Macrocyclic compounds as multi-target kinase inhibitors

Abstract

The present application relates to a macrocyclic compound as a multi-target kinase inhibitor, wherein the macrocyclic compound has a structure of formula (I). The macrocyclic compound has the activity of inhibiting multiple protein kinases such as HPK1, FLT3 and KDR, and can be used to treat diseases or conditions mediated by these protein kinases.

Description

Technical Field

[0001] The present invention relates to macrocyclic compounds, specifically to macrocyclic compounds with multi-target kinase inhibitory activity and pharmaceutical compositions thereof, and also to the use of such compounds and pharmaceutical compositions thereof in improving or treating diseases mediated by protein kinases, such as cancer. Background Art

[0002] Cancer, also known as malignant tumors, is a major threat to human health and life, with cancer morbidity and mortality rates continuing to rise. Tumor cell proliferation, metastasis, and apoptosis are closely linked to abnormalities in a series of signaling pathways. Immune evasion by tumor cells and the exhaustion or suppression of the host's immune function are also key factors in the development and progression of cancer. Therefore, inhibiting the activity of key kinases in tumor cell signaling pathways while simultaneously activating immune cells and enhancing anti-tumor immunity can effectively inhibit tumor cell growth and metastasis, offering promising therapeutic opportunities for cancer patients.

[0003] Hematopoietic progenitor kinase 1 (HPK1) is a serine / threonine kinase originally cloned from hematopoietic progenitor cells (Hu, MC et al., Genes Dev. 1996; 10:2251-2264; Keifer, F. et al., The EMBO Journal 1996; 15:7013-7025). It belongs to the mitogen-activated protein kinase (MAP4K) family. HPK1 is concentrated in lymphoid organs and tissues, such as the bone marrow, lymph nodes, and thymus, and is primarily expressed in immune cells (T cells, B cells, and dendritic cells) (Hu, MC et al., Genes Dev. 1996; 10:2251-2264).

[0004] Studies have shown that HPK1 is a negative regulator of the T cell receptor (TCR) signaling pathway. TCR signaling activates HPK1, which then phosphorylates SLP-76 at Ser376, promoting its binding to the 14-3-3 protein (Di Bartolo, V. et al., J. Exp. Med. 2007; 204: 681-691; Shui, J. et al., Nature Immuno. 2007; 8: 84-91). The SLP-76 / 14-3-3 interaction downregulates ERK signaling and calcium flux, triggering the ubiquitination of SLP-76 and the degradation of the SLP-76 complex, blocking the TCR activation pathway and thereby inhibiting T cell function (Lasserre, R. et al., J. Cell Biol. 2011; 195: 839-853).

[0005] In vivo experiments, HPK1 knockout mice showed enhanced T cell function under antigen stimulation, producing more cytokines such as IL-2 and IFN-γ (Shui, J. et al., Nature Immuno. 2007; 8: 84-91; Alzabin, S. et al., J. Immunol. 2009; 182: 6187-6194; Alzabin, S. et al., Cancer Immunol. Immunother. 2010; 59: 419-429). Further studies have shown that the negative regulation of HPK1 on immune cells depends on its kinase activity. Compared with wild-type mice, CD8 + Enhanced T cell function can clear chronic lymphocytic meningitis virus faster and better inhibit tumor growth (Hernandez, S. et al., Cell Reports 2018; 25: 80-94). In the Lewis lung cancer (LLC) model, HPK1 was transfected - / - Mice expressing HPK1 T cells showed stronger anti-tumor immune responses than wild-type mice (Sawasdikosol, S. et al., Immunol. Res. 2012; 54: 262-265). Studies have revealed that HPK1 has an effect on B cells (Sauer, K. et al., J. Biol. Chem. 2001; 276: 45207-45216; Tsuji, S. et al., J. Exp. Med. 2001; 194: 529-539; Wang, X. et al., J. Biol. Chem. 2012; 287: 34091-34100; S. et al., PLos One, 2010; 5: e12468), dendritic cells (Alzabin, S. et al., J. Immunol. 2009; 182: 6187-6194), NK cells and Treg cells also originate from their kinase activity (Liu, J. et al., PLos One, 2019; 14: e0212670).

[0006] Clinical studies have found that compared with healthy controls, HPK1 levels in tissues of patients with systemic lupus erythematosus (Zhang, Q. et al., J. Autoimmun., 2011; 37: 180-189) and psoriatic arthritis (Stoeckman, AK et al., Genes Immun. 2006; 7: 583-591; Baltiwalla, FM et al., Mol. Med. 2005; 11: 21-29) are significantly downregulated, indicating that HPK1 downregulation helps enhance autoimmune responses. On the other hand, upregulation of HPK1 levels has been observed in a variety of cancers, such as acute myeloid leukemia (Chen-Deutsch, X. et al., Leuk. Res. 2012; 36: 884-888; Chen-Deutsch, X. et al., Cell Cycle 2012; 11: 1364-1373), bladder urothelial carcinoma (Wang. Y et al., Mol. Med. Rep. 2012; 5: 260-265), extramammary Paget's disease (Qian, Y et al., Am J. Dermatopathol. 2011; 33: 681-686) and colon cancer (Yang, HS et al., Mol. Cell Biol. 2006; 26: 1297-1306).

[0007] Therefore, HPK1 is a promising potential target for the treatment of tumors and viral diseases, and the development of small molecule inhibitors of HPK1 kinase has important clinical prospects.

[0008] FMS-like tyrosine kinase 3 (FLT3) is a receptor tyrosine kinase expressed in hematopoietic progenitor cells and stem cells, playing an important role in the survival, proliferation, and differentiation of hematopoietic cells. Mutations and abnormal expression of FLT3 lead to hematologic diseases such as leukemia (Gilliland, DG et al., Blood, 2002; 100: 1532-1542; Stirewalt, DL et al., Nat. Rev. Cancer, 2003; 3: 650-665). Approximately 30% of adult acute myeloid leukemia (AML) patients harbor FLT3 gene mutations (Nakao, MS et al., Leukemia, 1996; 10: 1911-1918; Kottaridis PD, Blood, 2001; 98: 1742-1759). This is the most common gene mutation and a poor prognostic factor in patients (Abu-Duhier FM et al., British Journal of Haematology, 2000; 111: 190-195; Meshinchi S. et al., Clin. Cancer Res., 2009; 15: 4263-4269), making FLT3 an important target for the development of small molecule drugs to treat this type of tumor.

[0009] In 2017, Novartis (USA) approved midostaurin, the first small molecule FLT3 inhibitor, for combination therapy in treatment-naive FLT3-positive AML patients. Subsequently, Gilteritinib, developed by Astellas, and Quizartinib, developed by Daiichi Sankyo, were launched as monotherapy or in combination for the treatment of AML associated with abnormal FLT3 expression or mutation. Despite the availability of several drugs, small molecule FLT3 inhibitors as a treatment for AML remain problematic, including short-lived clinical effects and the development of drug resistance. Furthermore, clinical data suggest that FLT3 inhibitors can effectively eliminate blasts in patients' peripheral blood, but have less potent effects on the bone marrow (Bortheakur, G. et al., Haematologica, Jan. 2011;96:62-68). One possible reason for these issues is the presence of alternative signaling pathways. Therefore, the development of a new generation of FLT3 inhibitors, particularly small molecules capable of simultaneously inhibiting multiple signaling pathways, is needed to address unmet clinical needs.

[0010] Vascular endothelial growth factor receptor (VEGFR) belongs to the class III receptor tyrosine kinase family. VEGFR2 (KDR), also known as the kinase insert domain receptor (KDR), is widely distributed in vascular endothelial cells and plays a crucial role in inducing the growth, proliferation, and migration of neoplastic endothelial cells.

[0011] Currently, a variety of VEGFR2 inhibitors have been approved for the treatment of related tumors. In 2006, Sunitinib, a small molecule drug developed by Pfizer, was approved for marketing as a VEGFRs inhibitor for the treatment of gastrointestinal stromal tumors and metastatic renal cell carcinoma. As a multi-target inhibitor, Sunitinib not only effectively inhibits VEGFRs activity, but also has inhibitory effects on multiple kinases such as PDGFR and c-Kit. The multi-target inhibition effect makes the drug not only potentially applicable to multiple indications, but also can reduce drug resistance caused by activation of bypass signals. Since then, a number of multi-target small molecule inhibitors have been approved and have achieved great success, such as Sorafenib, Cabozantininb, Lenvatininb, etc.

[0012] Although some patent applications for HPK1 small molecule inhibitors have been disclosed, such as WO2018049152, WO2018049191, WO2018049200, WO2018049214, WO2018102366, WO2018183964, WO2019051199, WO2019090198, WO2019206049, WO2019238067, WO2020092528, WO2020103896, WO2021050964, WO2021213317, WO2022068848, WO2022089225, WO2022095904 and WO2022253328, there are still no drugs targeting HPK1 on the market. In particular, small molecule compounds that simultaneously inhibit HPK1 and other kinase targets are rarely reported, as exemplified by WO2020188467, WO2020235902, and WO2023278483. These small molecule compounds have the potential to directly inhibit tumor cell growth while simultaneously stimulating autoimmunity, thereby eliminating tumor cells. Therefore, the development of novel, highly active small molecule HPK1 inhibitors is urgently needed clinically. Summary of the Invention

[0013] The present application provides a macrocyclic compound of formula (I), or its isomers, pharmaceutically acceptable salts, polymorphs, isotope-labeled substances, active metabolites or prodrugs,

[0014]

[0015] in,

[0016] W is N or CH;

[0017] X is N, C-CN or carbonyl, and the dotted line between X and Y indicates the presence or absence of the double bond, wherein:

[0018] When X is N or C-CN, Y is C, Z is CH, and there is a double bond between X and Y;

[0019] When X is a carbonyl group, Y and Z are both N, and there is a single bond between X and Y;

[0020] Ring B is a benzene ring;

[0021] L is independently selected at each occurrence from -O-, -CH2-, -CH(R 1 )-、-C(R 1 )2-、 And one L connected to -O- is not -O-; and any two adjacent L are not -O- at the same time. And not at the same time m mutually connected Ls constitute a linker connecting -O- and the B ring;

[0022] R 1 Selected from F and C 1-3 alkyl;

[0023] R 2 Selected from H, F and C 1-3 alkoxy;

[0024] R 3 Selected from halogen, optionally substituted with 1, 2 or 3 R 31A C 1-3 Alkyl, optionally substituted with 1, 2 or 3 R 31A C 3-6 Cycloalkyl, C 2-6 Alkynyl and cyano groups;

[0025] R 4 Selected from H, F and C 1-3 alkoxy;

[0026] R 5 Selected from:

[0027] 1) H, F, and C 1-3 Alkoxy,

[0028] 2) optionally substituted with 1 or 2 R 51A phenyl and a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group contains 1 or 2 ring-forming heteroatoms selected from N, O or S,

[0029] 3) optionally substituted with 1, 2 or 3 R 51B A 5-7 membered monocyclic alicyclic group or a 7-10 membered bicyclic alicyclic group;

[0030] R 6 Selected from:

[0031] 1) H, F, and C 1-3 Alkoxy,

[0032] 2) One of the following structures, where the chemical bond ends are It indicates that it is connected to other atoms in the structure of formula (I) through this bond:

[0033]

[0034] And R 5 and R 6 Not H, F or C at the same time 1-3 alkoxy;

[0035] The condition is that when X is N, and (1) n is 0, R 4 and R 5 are all H, and L is independently selected from -O- or -CH2-, or, (2) n is 0, R 4 and R 5 Both are H, and -(L) m -for -(L) m -When connected to the B ring through the bond marked with "*", R 6 Select one of the following structures:

[0036]

[0037] R 31A Independently selected from F, hydroxy, C 1-3 Alkoxy, amino, C 1-3 Alkylamino, and cyano;

[0038] R 31B Selected from optionally substituted with 1, 2 or 3 R 311 C 1-3 Alkyl and cyclopropyl, where R 311 Selected from F or hydroxyl;

[0039] R 51A Independently selected from:

[0040] 1) Halogen, -OR a1 、-NR a1 R b1 、-NR a1 C(=O)R b1 、-NR e1 C(=O)NR a1 R b1 、-NR a1 S(=O)2R d1 、-NR e1 S(=O)2NR a1 R b1 、-C(=O)R a1 、-C(=O)OR a1 、-C(=O)NR a1 R b1 and cyano, and optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl,

[0041] 2) optionally substituted with 1, 2 or 3 R 52 5-6 membered aliphatic heterocyclic group;

[0042] R 51B Independently selected from:

[0043] 1) Oxo, F, hydroxyl, C 1-3 Alkoxy, cyclopropyloxy, amino, C 1-3 Alkylamino, cyclopropylamino, C 1-3 Alkyl, cyclopropyl and cyano, the C 1-3 Alkoxy, cyclopropyloxy, C 1-3 Alkylamino, C 1-3 Alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R G ;

[0044] 2) optionally substituted with 1, 2 or 3 R G A 4-6 membered saturated aliphatic heterocyclic group; or

[0045] 3) Two R attached to the same carbon atom 51B Together with the carbon atom,

[0046] R 52 Selected from R G 、C 1-3 Hydroxyalkyl and C 1-3 cyanoalkyl;

[0047] R a1 、R b1and R e1 Each independently selected from:

[0048] 1) Hydrogen;

[0049] 2) optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0050] or,

[0051] R attached to the same nitrogen atom a1 and R b1 Together with the nitrogen atom, it forms an optionally substituted 1 or 2 R G 3-6 membered aliphatic heterocyclic group;

[0052] R d1 independently selected from optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0053] R A Selected from H, optionally substituted with 1, 2 or 3 R A11 C 1-3 alkyl, Among them, R A11 is selected from fluoro, hydroxy or cyano;

[0054] R B Selected from H, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino, optionally substituted with 1, 2 or 3 R B11 C 1-3 Alkyl, cyclopropyl, cyano, Among them, R B11 is selected from fluorine or hydroxyl; wherein, The ring carbon atoms in are optionally substituted with 1, 2 or 3 C 1-3 Alkyl; The ring-forming carbon atoms in are optionally substituted with 1, 2 or 3 R G ; and, two R attached to the same carbon atom B Different from H, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino,

[0055] R G Selected from oxo, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Fluoroalkoxy, cyclopropyloxy, C 1-3 Alkyl, C1-3 Fluoroalkyl, cyclopropyl, and cyano groups;

[0056] E 1 Selected from -CH(R x )-、 where R X Selected from

[0057] E 2 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(R Y )-、 where R Y Selected from isopropyl,

[0058] E 3 Selected from -C(CH3)2-, -CH(CH2CN)-,

[0059] E 4 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(CH2CN)-,

[0060] E 5 Selected from -CH2-, -C(CH3)2-, -CF2-, where R Z Selected from H and C 1-3 alkyl;

[0061] The E ring is a 3-membered saturated alicyclic ring or a 4-6-membered saturated alicyclic heterocyclic ring, wherein the saturated alicyclic heterocyclic ring contains 1 or 2 ring-forming heteroatoms selected from N and O, and the ring-forming carbon atoms in the saturated alicyclic heterocyclic ring are optionally substituted with 1, 2 or 3 C 1-3 Alkyl, saturated aliphatic heterocyclic ring, if present, is optionally substituted with a ring nitrogen atom of R A ;

[0062] m is 4, 5 or 6;

[0063] n is 0, 1, or 2;

[0064] q is 1 or 2.

[0065] One embodiment of the present invention further provides a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, and a pharmaceutically acceptable carrier.

[0066] One embodiment of the present invention further provides a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, or the use of the above-mentioned pharmaceutical composition in preventing or treating diseases mediated by protein kinases.

[0067] One embodiment of the present invention further provides a use of the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, or the pharmaceutical composition in the preparation of a medicament for preventing or treating diseases mediated by protein kinases, wherein the diseases include one or more of tumors, myelodysplastic syndromes and diseases caused by viruses.

[0068] One embodiment of the present invention further provides a method for inhibiting HPK1 kinase activity, comprising administering to a subject a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof;

[0069] One embodiment of the present invention further provides a method for simultaneously inhibiting the activity of multiple protein kinases, which comprises administering to an individual a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof; the protein kinases are selected from one or more of HPK1, FLT3 and KDR.

[0070] One embodiment of the present invention further provides a method for treating a disease or condition mediated by protein kinase in a patient, comprising administering to the patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof.

[0071] The compounds of the present application have the effect of inhibiting the activities of multiple protein kinases, including HPK1, FLT3 and KDR, and can be used to treat diseases or conditions mediated by these protein kinases. DETAILED DESCRIPTION

[0072] The present invention will be further described in detail below through the following examples, through which the features and advantages of the present invention will become more clearly understood.

[0073] The word “exemplary” is used exclusively herein to mean “serving as an example, example, or illustration.” Any embodiment described herein as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments.

[0074] In addition, the technical features involved in different embodiments of the present application described below can be combined with each other as long as they do not conflict with each other.

[0075] definition

[0076] In the present invention, unless otherwise specified, the description or explanation of a group applies to all groups containing this group, whether used alone or in combination with other terms. For example, the description of alkyl applies to C 1-6 Alkyl, C 1-3 Alkyl, etc.; for C 1-6 The description or description of alkyl is applicable to "C 1-6 "alkoxy" etc., and the following definitions apply to the claims and the specification.

[0077] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition in each instance is independent. Thus, for example, if a group is substituted with 0-2 R's, the group may optionally be substituted with up to two R's, with each instance of R being an independent choice. Furthermore, combinations of substituents and / or their variants are permitted only if such combinations result in stable compounds. For example, when a structure contains multiple substituents represented by the same symbol, the substituents may be of the same or different types; for example, if the B ring group contains 2 R's .... 3 Substituents, these two R 3 Both may be methoxy groups, or one may be a methoxy group and the other a methyl group.

[0078] The term "optionally" or "optionally" means that the event or circumstance described subsequently may or may not occur, and the description includes both the occurrence of the event or circumstance and the non-occurrence of the event or circumstance. For example, "a chemical moiety optionally substituted with 1, 2, or 3 R substituents" means that the chemical moiety may not have substituents R, or the chemical moiety may have 1, 2, or 3 substituents R.

[0079] The terms "halo", "halogen" or "halogen atom" include fluorine, chlorine, bromine, iodine, in particular fluorine and chlorine.

[0080] The term "C m-n " group (wherein m and n are integers) indicates a range including the end point, which means that the corresponding group contains mn carbon atoms. For example, C 1-6 Alkyl refers to an alkyl group containing 1 to 6 carbon atoms, C 2-6 Alkenyl means an alkenyl group containing 2 to 6 carbon atoms.

[0081] The term "n-membered" (where n is an integer) generally describes the number of ring-forming atoms in a moiety, where the number of ring-forming atoms is n. "mn-membered" indicates an inclusive range, meaning that the corresponding ring structure contains mn ring-forming atoms. For example, piperidinyl is an example of a 6-membered heterocyclic group, and pyrazolyl is an example of a 5-membered heteroaromatic ring group.

[0082] The term "substituted" refers to an atom or group of atoms that formally replaces hydrogen as a "substituent" attached to another group. Unless otherwise indicated, the term "substituted" refers to any degree of substitution, as long as the substitution is allowed. The choice of substituent is independent, and substitution can be at any chemically accessible position. It should be understood that substitution on a given atom is limited by valence. It should be understood that substitution on a given atom produces a chemically stable molecule. A single divalent substituent (e.g., oxo) can replace two hydrogen atoms.

[0083] The "remainder of the compound" refers to the portion of the complete molecular structure excluding the substituent. The remainder of the compound is connected to the substituent through one or more unsaturated valences. The remainder of the compound may contain one or more "points of attachment," and two or more points of attachment may be on the same atom or on different atoms.

[0084] The term "alkyl" refers to a straight or branched saturated hydrocarbon group. An alkyl group is a group formed by losing a hydrogen atom from an alkane. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, 1,2,2-trimethylpropyl, etc. 1-3 Non-limiting examples of alkyl groups include, for example, methyl, ethyl, n-propyl, isopropyl, and the like.

[0085] The term "fluoroalkyl" refers to a group in which one or more hydrogen atoms in an alkyl group are replaced by fluorine atoms. n The "fluoroalkyl" group may have no less than 1 and no more than 2n+1 hydrogen atoms replaced by fluorine atoms. Examples of fluoroalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, and perfluorobutyl. 1-3 Non-limiting examples of fluoroalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, 1-fluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, perfluoroethyl, and the like.

[0086] The term "hydroxyalkyl" refers to a group in which one or more hydrogen atoms in an alkyl group are replaced by hydroxyl groups, and no more than one hydrogen atom on the same carbon atom can be replaced by hydroxyl groups. n The "hydroxyalkyl" group may have no fewer than 1 and no more than n hydrogen atoms replaced by hydroxy groups.1-3 Examples of hydroxyalkyl groups include, for example, hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 2-hydroxy-2-propyl, 2,3-dihydroxypropyl, and the like.

[0087] The term "cyanoalkyl" refers to a group in which one hydrogen atom of an alkyl group is replaced by a cyano group. 1-3 Examples of cyanoalkyl include cyanomethyl, 1-cyanoethyl, 2-cyanoethyl, 2-cyano-2-propyl and the like.

[0088] The term "alkynyl" refers to a straight or branched hydrocarbon group having one or more carbon-carbon triple bonds. An alkynyl group is a group formed by losing a hydrogen atom from an alkyne. 2-6 Examples of alkynyl groups include ethynyl, 1-propynyl, propargyl, 1-butynyl, but-2-yn-1-yl, but-3-yn-1-yl, but-3-en-1-ynyl, 3-methylpent-2-en-4-yn-1-yl, and the like.

[0089] The term "alkylene" refers to a divalent group formed by the simultaneous loss of two hydrogen atoms from a carbon atom of an alkane, wherein the two connecting valences may be concentrated on the same atom or the two connecting valences may be located on two atoms respectively. The two connecting valences may be located on the same atom of the rest of the compound or on two atoms respectively in the rest of the compound. Non-limiting examples of alkylene include, for example, methylene (-CH2- or =CH2), 1,1-ethylene (-CH(CH3)- or =CH-CH3), 1,2-ethylene (-CH2CH2-), butane-1,4-diyl, butane-1,3-diyl, 2,2-dimethyl-propane-1,3-diyl, and the like.

[0090] The term "alkoxy" refers to a group of the formula "-O-alkyl", wherein alkyl is as defined above. Non-limiting examples of alkoxy include, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, and n-hexoxy. 1-3 Examples of alkoxy groups include, for example, methoxy, ethoxy, n-propoxyethyl and isopropoxy.

[0091] The term "amino" refers to a group of formula -NH2. The term "alkylamino" includes groups of formula "-NH-alkyl" and groups of formula "-N(alkyl)2", wherein alkyl is as defined above. Examples of the group of formula "-NH-alkyl" include methylamino, ethylamino, isopropylamino, and n-hexylamino; examples of the group of formula "-N(alkyl)2" include dimethylamino, diethylamino, methylethylamino, methylisopropylamino, and ethyl-n-hexylamino. The term "C 1-3 "Alkylamino" includes the formula "-NH-C 1-3 Alkyl" and the group of formula "-N(C 1-3alkyl)2" groups, for example, methylamino, ethylamino, isopropylamino, dimethylamino, diethylamino, methylethylamino, methylisopropylamino, etc.

[0092] The term "cyclopropylamino" refers to a Dimethylamino, diethylamino, methylethylamino, methylisopropylamino

[0093] The term "carbonyl" refers to a group of formula -(C=O)-, which may also be written as -C(O)-.

[0094] The term "cyano" refers to a group of formula -C≡N, which may also be written as -CN.

[0095] The term "oxo" refers to an oxygen atom as a divalent substituent, which forms a carbonyl group when attached to a carbon atom, or forms a sulfoxide or sulfone group when attached to a heteroatom, or an N-oxide group, etc. In some embodiments, cycloalkyl and heterocyclyl groups may be optionally substituted with 1 or 2 oxo groups.

[0096] The term "cycloalkyl" includes monocyclic or polycyclic alicyclic and aromatic hydrocarbon groups. Monocyclic alicyclic hydrocarbon groups contain one hydrocarbon ring, including cyclized alkyl and alkenyl groups. Polycyclic alicyclic hydrocarbon groups contain two or more hydrocarbon rings, at least one of which contains an alicyclic hydrocarbon ring (including cyclized alkyl and alkenyl groups), and the other rings can be alicyclic hydrocarbon rings and / or aromatic hydrocarbon rings; any of which is connected to at least one other ring by a spiro ring (two rings share one ring atom) or a bridge ring (two rings share two or more ring atoms). Polycyclic cycloalkyl groups are connected to the rest of the compound through ring carbon atoms on the hydrocarbon ring. Examples of cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentenyl, cyclohexadienyl, bicyclo[3.1.0]hexyl, norbornyl, norpinyl, bicyclo[1.1.1]pentyl, 1H-inden-1-yl, 2,3-dihydro-1H-inden-2-yl, etc. When any of the rings contained in the cycloalkyl group is a saturated ring, the cycloalkyl group is a saturated cycloalkyl group, also known as a "cycloalkyl group". 3-6 Examples of the cycloalkyl group may include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

[0097] The term "aryl" refers to a monocyclic or polycyclic aromatic hydrocarbon group, for example, phenyl, naphthyl, etc.

[0098] The term "heterocyclyl" refers to a monocyclic or polycyclic group having at least one heteroatom ring member selected from oxygen, nitrogen, sulfur and phosphorus. A polycyclic heterocyclyl contains two or more ring structures, at least one of which has at least one heteroatom ring member selected from oxygen, nitrogen, sulfur and phosphorus, and the other ring structures may or may not have ring-forming heteroatoms; wherein any ring is connected to at least one other ring by a spirocycle (two rings share one ring atom) or a bridged ring (two rings share two or more ring atoms). The heterocyclyl may be connected to the rest of the compound by an optional ring-forming carbon atom, or may be connected to the rest of the compound by an optional ring-forming heteroatom. In one embodiment, any ring-forming carbon atom in the heterocyclyl may be substituted by an oxo group to form a carbonyl group. In one embodiment, any ring-forming nitrogen atom in the heterocyclyl may be an N-oxide. In one embodiment, any ring-forming nitrogen atom in the heterocyclyl may be a quaternary ammonium ion.

[0099] Heterocyclic groups include aromatic heterocyclic groups (ie, "heteroaryl groups") and "aliphatic heterocyclic groups."

[0100] "Heteroaryl" refers to a monocyclic or polycyclic aromatic heterocycle having at least one heteroatom ring member selected from oxygen, nitrogen, and sulfur. A heteroaryl group can be attached to the remainder of the compound through an optional carbon atom or through an optional heteroatom, provided that the valence of the carbon atom or heteroatom permits. In one embodiment, any ring-forming carbon atom in the heteroaryl moiety can be substituted with an oxo group to form a carbonyl group. In one embodiment, any ring-forming nitrogen atom in the heteroaryl moiety can be an N-oxide. In one embodiment, any ring-forming nitrogen atom in the heteroaryl moiety can be a quaternary ammonium ion. Examples of “heteroaryl” may include, for example, pyrrolyl (including pyrrol-1-yl, pyrrol-2-yl and pyrrol-3-yl), pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, tetrazolyl, pyridinyl, pyridin-2(1H)-on-1-yl, pyridin-4(1H)-on-1-yl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrazin-3(2H)-on-2-yl, 1,2,4-triazinyl, 1,3,5-triazinyl, indolyl, benzofuranyl, benzothiophenyl, indazolyl, benzimidazolyl, benzisothiazolyl, quinolyl, isoquinolyl, naphthyridinyl, imidazo[1,2-b]thiazolyl, purinyl and the like.

[0101] "Alicyclic groups" include monocyclic or polycyclic alicyclic groups. A monocyclic alicyclic group may contain no cyclic double bonds or may contain one or more cyclic double bonds. A polycyclic alicyclic group may contain at least one alicyclic ring structure, and the other rings may be alicyclic or aromatic rings; any ring structure in a polycyclic alicyclic group may contain no cyclic double bonds or may contain one or more cyclic double bonds. A "saturated alicyclic group" refers to an alicyclic group whose ring structure does not contain a cyclic double bond. A polycyclic alicyclic group may include a bicyclic alicyclic group, and may be a bridged ring or a spirocyclic group. Non-limiting examples of alicyclic groups include, for example, azetidinyl, oxetanyl, tetrahydropyrrolyl, tetrahydrofuranyl, 2-oxooxazolidinyl, piperidinyl, 3-oxopiperidinyl, piperazinyl, morpholinyl, azepanyl, 2-oxa-6-azaspiro[3.3]heptyl, 1,2,3,4-tetrahydroquinolinyl, and the like.

[0102] "Heterocyclylene" refers to a heterocyclyl group connected to two points of attachment to the rest of the compound via two valences. The two valences may be on the same ring atom of the heterocyclyl group or on two separate ring atoms of the heterocyclyl group; the two points of attachment may be on the same atom of the rest of the compound or on two separate atoms of the rest of the compound. Non-limiting examples of "heterocyclylene" include 1,1-(3-oxacyclobutane) and 1,3-(2-azacyclopentane).

[0103] The term "isomer" refers to an isomer resulting from the different spatial arrangements of atoms in a molecule. "Stereoisomers" of the compounds described herein refer to all stereoisomers. For example, when the compound has an asymmetric carbon atom, enantiomers and diastereomers are produced; when the compound has a carbon-carbon double bond, a carbon-nitrogen double bond, or a ring structure, cis-trans isomers are produced. Unless otherwise specified, the compounds described herein include all isomers thereof, such as optically active isomers, geometric isomers, rotational isomers, tautomers, and conformational isomers that can exist stably; and may exist as mixtures of isomers or as separated isomers.

[0104] Methods for preparing optically active products from optically inactive starting materials are known in the art, for example by resolution of racemic mixtures or by stereoselective synthesis.

[0105] The resolution of a racemic mixture of compounds can be carried out by any of a number of methods known in the art. One method includes fractional recrystallization using a chiral resolving acid that is an optically active, salt-forming organic acid. Suitable resolving agents for fractional recrystallization can be optically active acids such as D-tartaric acid, L-tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, L-camphorsulfonic acid, and the like; other suitable resolving agents for fractional recrystallization include, for example, stereoisomerically pure α-methylbenzylamine, 2-phenylglycinol, 1-cyclohexylethylamine, and the like.

[0106] The resolution method of the racemic mixture also includes, for example, converting to diastereomers by reaction with an appropriate optically active substance (e.g., chiral alcohol or Mosher's acyl chloride), separating the diastereomers and converting (e.g., hydrolyzing) them into corresponding single optical isomers. For example, eluting on a chromatographic column filled with an optically active resolving agent is implemented. Suitable chromatographic columns and elution solvent compositions can be determined by those skilled in the art.

[0107] The term "isotopically labeled" refers to a compound of the present invention in which one or more constituent atoms are replaced by a specific isotope thereof. For example, the isotopes in the compounds of the present invention may include various isotopes of elements such as H, C, N, O, F, P, S, Cl, and I, such as 2 H. 3 H. 13 C. 14 C. 15 N. 17 O. 18 O. 18 F. 30 P. 32 P. 35 S. 36 S. 123 I. 124 I and 125 I etc. The present invention includes various compounds that are isotopically labeled as defined. For example, radioactive isotopes (such as 3 H and 14 C), or those compounds in which non-radioactive isotopes such as 2 H and 13 C). Such isotope-labeled compounds are suitable for metabolic studies (using 14 C); reaction kinetic studies (using e.g. 2 H or 3 H); detection or imaging techniques, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug or substrate tissue distribution analysis; or radiotherapy applied to the patient, etc.

[0108] Specifically, 18 Compound F may be particularly desirable for PET or SPECT studies. Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by methods analogous to those described in the accompanying examples and preparations, using an appropriate isotopically labeled reagent in place of the unlabeled reagent previously employed.

[0109] In addition, the use of heavier isotopes, especially deuterium (i.e. 2 H or D) substitution may afford certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements or improved therapeutic index, and therefore may be preferred in some circumstances.

[0110] The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms which, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0111] "Pharmaceutically acceptable salts" refer to salts that retain the biological utility and properties of the compounds of the invention and generally do not have biologically or otherwise undesirable effects. In many cases, the compounds of the invention are capable of forming acid addition salts and / or base addition salts by virtue of the presence of amino and / or carboxyl groups or groups similar thereto.

[0112] Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids include, for example, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids include, for example, but not limited to, acetic acid, propionic acid, glycolic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethylsulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, ethanedisulfonic acid, sulfosalicylic acid, aspartic acid, glutamic acid, and the like.

[0113] Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases include, for example, but not limited to, basic compounds of sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium, and magnesium salts. Organic bases include primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like; examples include, but are not limited to, isopropylamine, benzylamine, choline, diethanolamine, diethylamine, dicyclohexylamine, lysine, arginine, meglumine, piperazine, and tromethamine.

[0114] All compounds and pharmaceutically acceptable salts thereof can be found together with other substances (e.g., solvents, including water and other solvents, etc.) (e.g., hydrates and solvates) or can be isolated. When in a solid state, the compounds described herein and their salts can exist in various forms, including hydrates and solvates. Hydrates and solvates of the compounds described herein and their salts include those in which the water and solvent can be isotopically substituted, such as D2O, methanol-d3, methanol-d4, acetone-d6, DMSO-d6. The presence of hydrates and solvates can be identified by those skilled in the art using means such as nuclear magnetic resonance (NMR).

[0115] The term "polymorph" refers to compounds of the present invention that exist in different crystalline forms, as well as in an amorphous form. Polymorphs of the compounds of the present invention and their salts also include mixtures of various crystalline forms, as well as mixtures of one or more crystalline forms with an amorphous form. The presence of polymorphs can be identified by those skilled in the art using methods such as X-ray diffraction.

[0116] Therefore, unless otherwise expressly stated, the compounds and salts thereof mentioned in this specification should be understood to cover any solid state form of the compounds.

[0117] The term "active metabolite" refers to an active derivative of a compound that is formed when the compound is metabolized.

[0118] "Pharmaceutically acceptable prodrugs" refer to any pharmaceutically acceptable ester, salt, amide, or other derivative of a compound of the invention that, upon administration to a recipient, is capable of providing, directly or indirectly, the compound of the invention or a pharmaceutically active metabolite or residue thereof. Particularly preferred derivatives or prodrugs are those that, when administered to a patient, can increase the bioavailability of the compound of the invention (e.g., by making an orally administered compound more readily absorbed into the bloodstream), or those that facilitate the delivery of the parent compound to a biological organ or site of action.

[0119] The term "pharmaceutical composition" refers to a biologically active compound optionally admixed with at least one pharmaceutically acceptable chemical component or agent, known as a "carrier," which facilitates the introduction of the compound into cells or tissues, including, but not limited to, stabilizers, diluents, suspending agents, thickeners, and / or excipients. Pharmaceutical compositions include, but are not limited to, tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (solid or dissolved in a liquid vehicle), ointments, soft and hard gelatin capsules, suppositories, transdermal patches, sterile injectable solutions, and sterile packaged powders.

[0120] The term "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antibacterial agents, antifungal agents), isotonic agents, absorption delaying agents, salts, preservatives, pharmaceutical stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavorings, dyes, and the like, and combinations thereof, as known to those skilled in the art. Unless any conventional carrier is incompatible with the active ingredient, its use in therapeutic or pharmaceutical compositions is contemplated.

[0121] The term "administration" or "administering" includes the introduction of the compound and its pharmaceutical composition into a subject to achieve its intended function. The route of administration depends on whether it is local or systemic treatment and the area to be treated. Examples of routes of administration that can be used include injection (including subcutaneous, intravenous, intraarterial, parenteral, intraperitoneal, intrathecal and other local injections), topical (including transdermal, epidermal, eye and mucous membranes, including intranasal, vaginal and rectal delivery), oral, inhalation or insufflation (intratracheal or intranasal, such as inhalation or insufflation of powders or aerosols, including by nebulizer), injection can be by way of a single bolus dose, or can be (for example) by way of infusion via a continuous infusion pump.

[0122] "Therapeutically effective amount" refers to the amount of the compound of the present invention that induces a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0123] "Subject" or "patient" refers to an individual suffering from a disease, disorder, condition, etc., including mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans; non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; other domesticated animals, such as rabbits, dogs, and cats; laboratory animals, including rodents, such as rats, mice, and guinea pigs. Examples of non-mammals include, but are not limited to, birds and fish.

[0124] Compound

[0125] In one embodiment, the present application relates to a macrocyclic compound of formula (I), or an isomer, pharmaceutically acceptable salt, polymorph, isotope label, active metabolite or prodrug thereof,

[0126]

[0127] in,

[0128] W is N or CH;

[0129] X is N, C-CN or carbonyl, and the dotted line between X and Y indicates the presence or absence of the double bond, wherein:

[0130] When X is N or C-CN, Y is C, Z is CH, and there is a double bond between X and Y;

[0131] When X is a carbonyl group, Y and Z are both N, and there is a single bond between X and Y;

[0132] Ring B is a benzene ring;

[0133] L is independently selected at each occurrence from -O-, -CH2-, -CH(R 1 )-、-C(R 1 )2-、 And one L connected to -O- is not -O-; and any two adjacent L are not -O- at the same time, and are not And not at the same time m mutually connected Ls constitute a linker connecting -O- and the B ring;

[0134] R 1 Selected from F and C 1-3 alkyl;

[0135] R 2 Selected from H, F and C 1-3 alkoxy;

[0136] R 3 Selected from halogen, optionally substituted with 1, 2 or 3 R 31A C 1-3 Alkyl, optionally substituted with 1, 2 or 3 R 31A C 3-6 Cycloalkyl, C 2-6 Alkynyl and cyano groups;

[0137] R 4 Selected from H, F and C 1-3 alkoxy;

[0138] R 5 Selected from:

[0139] 1) H, F, and C 1-3 Alkoxy,

[0140] 2) optionally substituted with 1 or 2 R 51A phenyl and a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group contains 1 or 2 ring-forming heteroatoms selected from N, O or S,

[0141] 3) optionally substituted with 1, 2 or 3 R 51B A 5-7 membered monocyclic alicyclic group or a 7-10 membered bicyclic alicyclic group;

[0142] R 6 Selected from:

[0143] 1) H, F, and C 1-3 Alkoxy,

[0144] 2) One of the following structures, where the chemical bond ends are It indicates that it is connected to other atoms in the structure of formula (I) through this bond:

[0145]

[0146] And R 5 and R 6 Not H, F or C at the same time 1-3 alkoxy;

[0147] The condition is that when X is N, and (1) n is 0, R 4 and R 5 are all H, and L is independently selected from -O- or -CH2-, or, (2) n is 0, R 4 and R 5 Both are H, and -(L) m -for -(L) m -When connected to the B ring through the bond marked with "*", R 6 Select one of the following structures:

[0148]

[0149] R 31A Independently selected from F, hydroxy, C 1-3 Alkoxy, amino, C 1-3 Alkylamino, and cyano;

[0150] R 31B Selected from optionally substituted with 1, 2 or 3 R 311 C 1-3 Alkyl and cyclopropyl, where R 311 Selected from F or hydroxyl;

[0151] R 51A Independently selected from:

[0152] 1) Halogen, -OR a1 、-NR a1 R b1 、-NR a1 C(=O)R b1 、-NR e1 C(=O)NR a1 R b1 、-NR a1 S(=O)2R d1 、-NRe1 S(=O)2NR a1 R b1 、-C(=O)R a1 、-C(=O)OR a1 、-C(=O)NR a1 R b1 and cyano, and optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl,

[0153] 2) optionally substituted with 1, 2 or 3 R 52 5-6 membered aliphatic heterocyclic group;

[0154] R 51B Independently selected from:

[0155] 1) Oxo, F, hydroxyl, C 1-3 Alkoxy, cyclopropyloxy, amino, C 1-3 Alkylamino, cyclopropylamino, C 1-3 Alkyl, cyclopropyl and cyano, the C 1-3 Alkoxy, cyclopropyloxy, C 1-3 Alkylamino, C 1-3 Alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R G ;

[0156] 2) optionally substituted with 1, 2 or 3 R G A 4-6 membered saturated aliphatic heterocyclic group; or

[0157] 3) Two R attached to the same carbon atom 51B Together with the carbon atom,

[0158] R 52 Selected from R G 、C 1-3 Hydroxyalkyl and C 1-3 cyanoalkyl;

[0159] R a1 、R b1 and R e1 Each independently selected from:

[0160] 1) Hydrogen;

[0161] 2) optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0162] or,

[0163] R attached to the same nitrogen atom a1and R b1 Together with the nitrogen atom, it forms an optionally substituted 1 or 2 R G 3-6 membered aliphatic heterocyclic group;

[0164] R d1 independently selected from optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0165] R A Selected from H, optionally substituted with 1, 2 or 3 R A11 C 1-3 alkyl, Among them, R A11 is selected from fluoro, hydroxy or cyano;

[0166] R B Selected from H, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino, optionally substituted with 1, 2 or 3 R B11 C 1-3 Alkyl, cyclopropyl, cyano, Among them, R B11 is selected from fluorine or hydroxyl; wherein, The ring carbon atoms in are optionally substituted with 1, 2 or 3 C 1-3 Alkyl; The ring-forming carbon atoms in are optionally substituted with 1, 2 or 3 R G ; and, two R attached to the same carbon atom B Different from H, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino,

[0167] R G Selected from oxo, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Fluoroalkoxy, cyclopropyloxy, C 1-3 Alkyl, C 1-3 Fluoroalkyl, cyclopropyl, and cyano groups;

[0168] E 1 Selected from -CH(R x )-、 where R X Selected from

[0169] E 2Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(R Y )-、 where R Y Selected from isopropyl,

[0170] E 3 Selected from -C(CH3)2-, -CH(CH2CN)-,

[0171] E 4 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(CH2CN)-,

[0172] E 5 Selected from -CH2-, -C(CH3)2-, -CF2-, where R Z Selected from H and C 1-3 alkyl;

[0173] The E ring is a 3-membered saturated alicyclic ring or a 4-6-membered saturated alicyclic heterocyclic ring, wherein the saturated alicyclic heterocyclic ring contains 1 or 2 ring-forming heteroatoms selected from N and O, and the ring-forming carbon atoms in the saturated alicyclic heterocyclic ring are optionally substituted with 1, 2 or 3 C 1-3 Alkyl, saturated aliphatic heterocyclic ring, if present, is optionally substituted with a ring nitrogen atom of R A ;

[0174] m is 4, 5 or 6;

[0175] n is 0, 1, or 2;

[0176] q is 1 or 2.

[0177] In one embodiment, W is selected from CH.

[0178] In one embodiment, W is selected from N.

[0179] In one embodiment, each L is independently selected from -O-, -CH2-, -C(CH3)2-, -CF2- and And one L connected to -O- is not -O-; and any two adjacent L are not simultaneously -O-, not simultaneously -C(CH3)2-, and not simultaneously

[0180] In one embodiment, each L is independently selected from -O-, -CH2- and And one L connected to -O- is not -O-; and any two adjacent Ls are not -O- at the same time, and are not

[0181] In one embodiment, each L is independently selected from -O- and -CH2-, and one L connected to -O- is not -O-, and any two adjacent Ls are not simultaneously -O-.

[0182] In one embodiment, R 1 Selected from F and methyl.

[0183] In one embodiment, R 1 Selected from F.

[0184] In one embodiment, R 1 Selected from methyl.

[0185] In one embodiment, -(L) m - is selected from one of the following structures and is connected to the B ring via a bond marked with "*",

[0186]

[0187] In one embodiment, -(L) m - one selected from the following structures,

[0188]

[0189] In one embodiment, -(L) m - one selected from the following structures,

[0190]

[0191] In one embodiment, -(L) m - one selected from the following structures,

[0192]

[0193] In one embodiment, -(L) m - one selected from the following structures,

[0194] And preferably,

[0195]

[0196] In one embodiment, R 2 Selected from F and C 1-3 Alkoxy groups, such as methoxy groups.

[0197] In one embodiment, R 2 Selected from F.

[0198] In one embodiment, R 2 Selected from H.

[0199] In one embodiment, there is only one R 3 , and is in R 2 Ortho position, R 3 is selected from F, Cl, methyl or cyclopropyl; and preferably is F and Cl.

[0200] In one embodiment, there is only one R 3 , and is in R 2 The meta position, R 3 is selected from F, Cl, methyl, difluoromethyl, trifluoromethyl, ethyl, isopropyl, cyclopropyl, ethynyl and cyano, wherein the methyl, ethyl, isopropyl and cyclopropyl are unsubstituted or substituted with one selected from hydroxy, C 1-3 Alkoxy, amino, C 1-3 Alkylamino, or cyano substituents; the ethynyl group is unsubstituted or substituted with one selected from methyl, cyclopropyl, substituents.

[0201] In one embodiment, there is only one R 3 , and is in R 2 The meta position, R 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are unsubstituted or substituted with one selected from hydroxy, methoxy, methylamino, dimethylamino or substituents.

[0202] In one embodiment, R 31A Selected from hydroxyl, C 13 Alkoxy, amino, C 13 Alkylamino and and preferably hydroxy, methoxy, methylamino, dimethylamino and

[0203] In one embodiment, R 31B Selected from methyl, cyclopropyl,

[0204] In one embodiment, there is only one R 3 , and is in R 2 The meta position, R 3 is selected from methyl, ethyl, ethynyl and cyano, wherein the methyl and ethyl groups are unsubstituted or substituted with one selected from hydroxy, methoxy, amino, methylamino, dimethylamino or Substituents;

[0205] In one embodiment, the B ring is excluding L and R 2 In addition, it does not contain other substituents, that is, there is no R 3 (n is 0).

[0206] In one embodiment, R 4 Selected from H and F.

[0207] In one embodiment, R 4 Selected from methoxy.

[0208] In one embodiment, R 4 Selected from F.

[0209] In one embodiment, R 4 Selected from H.

[0210] In one embodiment, R 5 Selected from H and F.

[0211] In one embodiment, R 5 Selected from F.

[0212] In one embodiment, R 5 Selected from H.

[0213] In one embodiment, X=N, W=CH, and the compound has the structure of formula (II-1)

[0214] in,

[0215] R 2 Selected from H and F;

[0216] R 3 There is only one, and R 3 In R 2 Ortho position, R 3 is selected from F, Cl, methyl or cyclopropyl,

[0217] R 4 and R 5 Selected from H, F and C 1-3 Alkoxy such as methoxy, or R 4 and R 5 All are H;

[0218] R 6 Select one of the following structures,

[0219]

[0220] Among them, E 2 、R B 、R A 、E 4 、E 5 , E ring and q are as defined above.

[0221] Preferably, R 6 Select one of the following structures:

[0222]

[0223] In one embodiment, the compound has the structure of formula (II-1),

[0224] in,

[0225] R 2 Selected from H and F;

[0226] R 3 There is only one, and R 3 In R 2 The meta position, R 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are unsubstituted or substituted with one selected from hydroxy, methoxy, methylamino, dimethylamino or Substituents;

[0227] R 4 and R 5 Selected from H, F and C 1-3 Alkoxy groups, such as methoxy,

[0228] R 6 Select one of the following structures,

[0229]

[0230] Among them, E 2 、R B 、R A 、E 4 、E 5 , E ring and q are as defined above.

[0231] In one embodiment, in the case where the above compound has the structure of formula (II-1), R 4 is selected from F and methoxy, and is preferably F,

[0232] R 5 is selected from H, F and methoxy, and is preferably H.

[0233] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein R 4 is selected from H, F and methoxy, and is preferably H,

[0234] R 5 It is selected from F and methoxy, and is preferably F.

[0235] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein R 4 and R 5There is only one selected from F and methoxy, or R 4 and R 5 All selected from F.

[0236] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein,

[0237] Each L is not all selected from -O- or -CH2-, and -(L) m -Not for

[0238] And - (L) m -Preferably the following structure:

[0239]

[0240] In one embodiment, in the case where the above compound has the structure of formula (II-1),

[0241] n is 0, and R 4 and R 5 Not all H,

[0242] Or, n is 1, R 3 In R 2 Ortho or meta position,

[0243] -(L) m -Selected from one of the following structures:

[0244]

[0245] And preferably one of the following structures:

[0246]

[0247] In one embodiment, in the case where the above compound has the structure of formula (II-1), R 4 is selected from H, F and methoxy; and preferably H,

[0248] R 6 is selected from H, F and methoxy; and preferably H,

[0249] R 5 Selected from:

[0250] 1) unsubstituted or optionally substituted with 1 or 2 R 51A phenyl and a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group contains 1 or 2 ring-forming heteroatoms selected from N, O or S,

[0251] 2) unsubstituted or optionally substituted with 1, 2 or 3 R 51BThe 5-7 membered monocyclic alicyclic group or the 7-10 membered bicyclic alicyclic group may be a bridged ring or a spiro ring.

[0252] In one embodiment, the compound has the structure of formula (IIa),

[0253]

[0254] Among them, -(L) m -Selected from one of the following structures:

[0255]

[0256] R 2 Selected from H and F;

[0257] R 3 Located in R 2 Ortho or meta, where (1) when R 3 Located in R 2 When the adjacent position of R 3 is selected from F, Cl, methyl or cyclopropyl; (2) when R 3 Located in R 2 When the meta position is 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are optionally substituted with one selected from hydroxy, methoxy, methylamino, dimethylamino or Substituents;

[0258] R 4 and R 5 There is only one selected from F and C 1-3 Alkoxy, or R 4 and R 5 All are H,

[0259] R 6 Select one of the following structures:

[0260] Among them, R A Selected from C 1-3 Alkyl, C 1-3 Fluorinated alkyl and C 1-3 hydroxyalkyl;

[0261] R B Selected from H, hydroxyl, C 1-3 Hydroxyalkyl, described The ring carbon atoms in are optionally substituted with 1, 2 or 3 hydroxyl or methyl groups; and the two R B Different from H, hydroxyl,

[0262] E 4 is selected from carbonyl and -CH2-;

[0263] The E ring is selected from one of the following structures:

[0264] wherein the ring-forming carbon atoms are unsubstituted;

[0265] q is selected from 1 and 2.

[0266] In one embodiment, the compound has the structure of formula (IIa),

[0267]

[0268] Among them, -(L) m -Selected from one of the following structures:

[0269]

[0270] R 2 Selected from H and F;

[0271] R 3 Located in R 2 Ortho or meta, where (1) when R 3 Located in R 2 When the adjacent position of R 3 is selected from F, Cl, methyl or cyclopropyl, (2) when R 3 Located in R 2 When the meta position is 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are optionally substituted with one selected from hydroxy, methoxy, methylamino, dimethylamino or Substituents;

[0272] R 4 and R 6 There is only one selected from F, or R 4 and R 6 All are H,

[0273] R 5 Selected from:

[0274] 1) optionally substituted with 1 or 2 R 51A pyridyl, imidazolyl and pyrazolyl,

[0275] 2) optionally substituted with 1, 2 or 3 R 51B The 5-6 membered monocyclic alicyclic group is preferably selected from:

[0276] in,

[0277] R 51A Selected from halogen, -OR a1 、-NR a1 R b1 、-NR a1 C(=O)R b1 、-NR a1 S(=O)2R d1 、

[0278] -C(=O)NR a1 R b1 、C 1-3 Alkyl, C 1-3 Fluorinated alkyl, C 1-3 Hydroxyalkyl, C 1-3 cyanoalkyl, cyclopropyl, and cyano groups;

[0279] R 51B Selected from:

[0280] 1) Oxo, F, hydroxy, methoxy, trifluoromethoxy, methylamino, dimethylamino, cyclopropylamino, methyl, difluoromethyl, trifluoromethyl, isopropyl, cyclopropyl, cyano,

[0281] or,

[0282] 2) Two R attached to the same carbon atom 51B Together with the carbon atom,

[0283] Among them, R a1 and R b1 Each independently selected from:

[0284] 1) Hydrogen;

[0285] 2) optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0286] or,

[0287] R attached to the same nitrogen atom a1 and R b1 Together with the nitrogen atom, it forms an optionally substituted 1 or 2 R G 3-6 membered aliphatic heterocyclic group;

[0288] R d1 independently selected from optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0289] R G Selected from oxo, F, hydroxyl, C1-3 Alkoxy, C 1-3 Fluoroalkoxy, cyclopropyloxy, C 1-3 Alkyl, C 1-3 Fluoroalkyl, cyclopropyl and cyano.

[0290] In one embodiment, in the case where the above compound has the structure of formula (IIa), R 5 Select one of the following structures:

[0291]

[0292] In one embodiment, X=C-CN, W=CH, and the compound has the structure of formula (II-2)

[0293]

[0294] Among them, R 2 Selected from H and F;

[0295] n is 1, that is, R 3 There is only one and it is in R 2 Ortho position, R 3 is selected from F, Cl, methyl or cyclopropyl;

[0296] R 4 and R 5 Selected from H, F and C 1-3 Alkyl groups such as methoxy,

[0297] R 6 Select one of the following structures,

[0298]

[0299] Among them, E 2 、R B 、R A 、E 4 、E 5 , E ring and q are as defined above.

[0300] In one embodiment, the compound has the structure of formula (II-2),

[0301]

[0302] Among them, R 2 Selected from H and F;

[0303] n is 1, that is, R 3 There is only one and it is in R 2 The meta position, R 3is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are unsubstituted or optionally substituted with one group selected from hydroxy, methoxy, methylamino, dimethylamino or Substituents;

[0304] R 4 and R 5 is selected from H, F and methoxy;

[0305] R 6 Select one of the following structures,

[0306]

[0307] Among them, E 2 、R B 、R A 、E 4 、E 5 , E ring and q are as defined above.

[0308] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein n is 1, that is, there is only one R 3 ,

[0309] When R 3 In R 2 When the adjacent position of R 3 is selected from F, Cl, methyl or cyclopropyl,

[0310] Or, R 3 In R 2 The meta position, R 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are unsubstituted or optionally substituted with one group selected from hydroxy, methoxy, methylamino, dimethylamino or Substituents;

[0311] R 4 and R 5 All selected from H.

[0312] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein R 4 and R 5 There is only one selected from F and methoxy, or R 4 and R 5 All selected from F.

[0313] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein R 4 and R 5 All selected from H.

[0314] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein -(L) m -Selected from one of the following structures:

[0315]

[0316] In one embodiment, in the case where the above compound has a structure of formula (II-1), wherein,

[0317] R 4 is selected from H, F and methoxy; and preferably H,

[0318] R 6 is selected from H, F and methoxy; and preferably H,

[0319] R 5 Selected from:

[0320] 1) unsubstituted or optionally substituted with 1 or 2 R 51A phenyl and a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group contains 1 or 2 ring-forming heteroatoms selected from N, O or S,

[0321] 2) unsubstituted or optionally substituted with 1, 2 or 3 R 51B The 5-7 membered monocyclic alicyclic group or the 7-10 membered bicyclic alicyclic group may be a bridged ring or a spiro ring.

[0322] In one embodiment, the compound has the structure of formula (IIb),

[0323]

[0324] Among them, -(L) m -Selected from one of the following structures:

[0325]

[0326] R 2 Selected from H and F;

[0327] R 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are optionally substituted with hydroxy, methoxy, methylamino, dimethylamino or

[0328] R 4 Selected from H, F and C 1-3 Alkoxy, R 5 Selected from H, F and C 1-3 Alkoxy, and R 4 and R 5 There is only one selected from F and C1-3 Alkoxy, or R 4 and R 5 All are H,

[0329] R 6 Select one of the following structures:

[0330] in,

[0331] R A Selected from C 1-3 Alkyl, C 1-3 Fluorinated alkyl and C 1-3 hydroxyalkyl;

[0332] R B Selected from H, hydroxyl, C 1-3 Hydroxyalkyl, described The ring carbon atoms in are optionally substituted with 1, 2 or 3 hydroxyl or methyl groups; and the two R B Different from H, hydroxyl,

[0333] E 4 is selected from carbonyl and -CH2-;

[0334] The E ring is selected from one of the following structures:

[0335] wherein the ring-forming carbon atoms are unsubstituted;

[0336] q is selected from 1 and 2.

[0337] In one embodiment, the compound has the structure of formula (IIb),

[0338]

[0339] Among them, -(L) m -Selected from one of the following structures:

[0340]

[0341] R 2 Selected from H and F;

[0342] R 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are optionally substituted with hydroxy, methoxy, methylamino, dimethylamino or

[0343] R 4 Selected from H, F and C 1-3 Alkoxy, R 6Selected from H, F and C 1-3 Alkoxy; and R 4 and R 6 There is only one selected from F, or R 4 and R 6 All are H,

[0344] R 5 Selected from:

[0345] 1) optionally substituted with 1 or 2 R 51A pyridyl, imidazolyl and pyrazolyl,

[0346] 2) optionally substituted with 1, 2 or 3 R 51B The 5-6 membered monocyclic alicyclic group is preferably selected from:

[0347] in,

[0348] R 51A Selected from halogen, -OR a1 、-NR a1 R b1 、-NR a1 C(=O)R b1 、-NR a1 S(=O)2R d1 、

[0349] -C(=O)NR a1 R b1 、C 1-3 Alkyl, C 1-3 Fluorinated alkyl, C 1-3 Hydroxyalkyl, C 1-3 cyanoalkyl, cyclopropyl, and cyano;

[0350] R 51B Selected from:

[0351] 1) Oxo, F, hydroxy, methoxy, trifluoromethoxy, methylamino, dimethylamino, cyclopropylamino, methyl, difluoromethyl, trifluoromethyl, isopropyl, cyclopropyl, cyano, or,

[0352] 2) Two R attached to the same carbon atom 51B Together with the carbon atom,

[0353] Among them, R a1 and R b1 Each independently selected from:

[0354] 1) Hydrogen;

[0355] 2) optionally substituted with 1, 2 or 3 RG C 1-3 Alkyl and cyclopropyl;

[0356] or,

[0357] R attached to the same nitrogen atom a1 and R b1 Together with the nitrogen atom, it forms an optionally substituted 1 or 2 R G 3-6 membered aliphatic heterocyclic group;

[0358] R d1 independently selected from optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0359] R G Selected from oxo, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Fluoroalkoxy, cyclopropyloxy, C 1-3 Alkyl, C 1-3 Fluoroalkyl, cyclopropyl and cyano.

[0360] In one embodiment, the compound has the structure of formula (IIc),

[0361]

[0362] Among them, R 2 Selected from H and F;

[0363] -(L) m -Selected from one of the following structures:

[0364]

[0365] R 3 Located in R 2 Ortho or meta, where (1) when R 3 Located in R 2 When the adjacent position of R 3 Selected from F and Cl; (2) when R 3 Located in R 2 When the meta position is 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are optionally substituted with hydroxy, methoxy, methylamino, dimethylamino or

[0366] R 4 and R 5 All are H;

[0367] R 6 Select one of the following structures:

[0368] Among them, R A Selected from C 1-3 Alkyl, C 1-3 Fluorinated alkyl and C 1-3 hydroxyalkyl;

[0369] R B Selected from H, hydroxyl, C 1-3 Hydroxyalkyl, described The ring carbon atoms in are optionally substituted with 1, 2 or 3 hydroxyl or methyl groups; and the two R B Different from H, hydroxyl,

[0370] E 4 is selected from carbonyl and -CH2-;

[0371] The E ring is selected from one of the following structures:

[0372] wherein the ring-forming carbon atoms are unsubstituted;

[0373] q is selected from 1 and 2.

[0374] In one embodiment, X=N, W=CH, R 2 Selected from F, n is 0, the compound has the structure of formula (IIIa)

[0375]

[0376] Among them, R 4 is selected from H, F and methoxy, R 5 is selected from H, F and methoxy; and R 4 and R 5 There is only one selected from F and methoxy, or R 4 and R 5 All selected from F,

[0377] R 6 Select one of the following structures,

[0378]

[0379] Among them, E 2 、R B 、R A 、E 4 、E 5 , E ring and q are as defined above.

[0380] In one embodiment, in the case where the above compound has a structure of formula (IIIa), wherein R 4Selected from F, R 5 Selected from H.

[0381] In one embodiment, in the case where the above compound has a structure of formula (IIIa), wherein R 4 Selected from H, R 5 Selected from F.

[0382] In one embodiment, in the case where the above compound has a structure of formula (IIIa), wherein -(L) m -Selected from one of the following structures:

[0383]

[0384] R 4 Selected from F, R 5 Selected from H.

[0385] In one embodiment, in the case where the above compound has a structure of formula (IIIa), wherein -(L) m -Selected from one of the following structures:

[0386]

[0387] R 4 Selected from H, R 5 Selected from F.

[0388] In one embodiment, in the case where the above compound has a structure of formula (IIIa), wherein -(L) m -Selected from one of the following structures:

[0389]

[0390] R 4 and R 5 All selected from H.

[0391] In one embodiment, the compound has the structure of formula (IIIa),

[0392]

[0393] wherein each L is independently selected from -O- or -CH2-, or -(L) m -Selected

[0394] R 4 and R 5 All are selected from H,

[0395] R 6 Select one of the following structures

[0396]

[0397] Among them, E 1 、R B 、R A 、E 3 、E 5 , E ring and q are as defined above.

[0398] In one embodiment, in the case where the above compound has a structure of formula (IIIa), wherein -(L) m -Selected from one of the following structures:

[0399]

[0400] R 4 and R 5 All selected from H.

[0401] In one embodiment, in the case where the above compound has a structure of formula (IIIa), wherein,

[0402] R 4 and R 6 All selected from H,

[0403] R 5 Selected from:

[0404] 1) unsubstituted or optionally substituted with 1 or 2 R 51A pyridyl, imidazolyl and pyrazolyl,

[0405] 2) unsubstituted or optionally substituted with 1, 2 or 3 R 51B A 5-7 membered monocyclic aliphatic heterocyclic group.

[0406] In one embodiment, X═C-CN, W═CH, R 2 is selected from F, n is 0, and the compound has the structure of formula (IIIb)

[0407]

[0408] Among them, R 4 is selected from H, F and methoxy, R 5 is selected from H, F and methoxy; and R 4 and R 5 There is only one selected from F and methoxy, or R 4 and R 5 All selected from F,

[0409] R 6 Select one of the following structures,

[0410]

[0411] Among them, E 2 、R B 、R A 、E 4 、E 5 , E ring and q are as defined above.

[0412] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein,

[0413] R 4 is selected from H, F and methoxy, R 5 is selected from H, F and methoxy; and R 4 and R 5 There is only one selected from F and methoxy,

[0414] -(L) m -Selected from one of the following structures:

[0415]

[0416] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein R 4 Selected from F, R 5 Selected from H.

[0417] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein,

[0418] R 4 Selected from H, R 5 Selected from F.

[0419] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein R 4 and R 5 All selected from H.

[0420] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein -(L) m -Selected from one of the following structures:

[0421]

[0422] R 4 Selected from F, R 5 Selected from H.

[0423] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein,

[0424] -(L) m -Selected from one of the following structures:

[0425]

[0426] R 4 Selected from H, R 5 Selected from F.

[0427] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein -(L) m -Selected from one of the following structures:

[0428]

[0429] R 4 and R 5 All selected from H.

[0430] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein,

[0431] -(L) m -Selected from one of the following structures:

[0432]

[0433] R 4 and R 5 All selected from H.

[0434] In one embodiment, in the case where the above compound has a structure of formula (IIIb), wherein R 4 and R 6 All selected from H,

[0435] R 5 Selected from:

[0436] 1) unsubstituted or optionally substituted with 1 or 2 R 51A pyridyl, imidazolyl and pyrazolyl,

[0437] 2) unsubstituted or optionally substituted with 1, 2 or 3 R 51B A 5-7 membered monocyclic aliphatic heterocyclic group.

[0438] In one embodiment, X = carbonyl, W = CH, R 2 Selected from F, n is 0, the compound has the structure of formula (IIIc)

[0439]

[0440] Among them, R 4 is selected from H, F and methoxy, R 5 is selected from H, F and methoxy; R 4 and R 5There is only one selected from F and methoxy, or R 4 and R 5 All selected from F,

[0441] R 6 Select one of the following structures,

[0442]

[0443] Among them, E 2 、R B 、R A 、E 4 、E 5 , E ring and q are as defined above.

[0444] In one embodiment, in the case where the above compound has a structure of formula (IIIc), wherein R 4 Selected from F, R 5 Selected from H.

[0445] In one embodiment, in the case where the above compound has a structure of formula (IIIc), wherein R 4 Selected from H, R 5 Selected from F.

[0446] In one embodiment, in the case where the above compound has a structure of formula (IIIc), wherein,

[0447] R 4 and R 5 All selected from H.

[0448] In one embodiment, in the case where the above compound has a structure of formula (IIIc), wherein,

[0449] -(L) m -Selected from one of the following structures:

[0450]

[0451] R 4 Selected from F, R 5 Selected from H.

[0452] In one embodiment, in the case where the above compound has a structure of formula (IIIc), wherein,

[0453] -(L) m -Selected from one of the following structures:

[0454]

[0455] R 4 Selected from H, R 5 Selected from F.

[0456] In one embodiment, in the case where the above compound has a structure of formula (IIIc), wherein,

[0457] -(L) m -Selected from one of the following structures:

[0458]

[0459] R 4 and R 5 All selected from H.

[0460] In one embodiment, the compound has the structure of formula (IIIa),

[0461]

[0462] Among them, -(L) m -Selected from one of the following structures:

[0463] in,

[0464] (1) When R 4 and R 5 There is only one selected from F and C 1-3 Alkoxy, the other is H,

[0465] R 6 Select one of the following structures:

[0466]

[0467] (2) When R 4 and R 6 When both are H,

[0468] R 5 Selected from:

[0469] 1) optionally substituted with 1 or 2 R 51A substituted pyridyl, imidazolyl and pyrazolyl groups,

[0470] 2) optionally substituted with 1, 2 or 3 R 51B The following ring systems:

[0471] in,

[0472] R 51A independently selected from F, Cl, methoxy, methyl, cyclopropyl and cyano;

[0473] R 51B Selected from:

[0474] 1) F, hydroxy, methyl, isopropyl and cyano, or

[0475] 2) Two R attached to the same carbon atom 51B Together with the carbon atom,

[0476] In one embodiment, the compound has the structure of formula (IIIa),

[0477]

[0478] Among them, -(L) m -Selected from one of the following structures:

[0479] in,

[0480] R 4 Selected from H, F and C 1-3 Alkoxy, R 5 Selected from H, F and C 1-3 Alkoxy; and R 4 and R 5 There is only one selected from F and C 1-3 Alkoxy, or R 4 and R 5 All are H,

[0481] R 6 Select one of the following structures:

[0482]

[0483] In one embodiment, the compound has the structure of formula (IIIa),

[0484]

[0485] Among them, -(L) m -Selected from one of the following structures:

[0486]

[0487] R 4 and R 5 All are H,

[0488] R 6 Select one of the following structures:

[0489] in,

[0490] R B Selected from H, hydroxyl, C 1-3 Hydroxyalkyl, described The ring carbon atoms in are optionally substituted with 1, 2 or 3 hydroxyl or methyl groups; and the two R B Different from H, hydroxyl,

[0491] The E ring is selected from one of the following structures:

[0492] wherein the ring-forming carbon atoms are unsubstituted;

[0493] q is selected from 1 and 2.

[0494] In one embodiment, the compound has the structure of formula (IIIa),

[0495]

[0496] Among them, -(L) m -Selected from one of the following structures:

[0497]

[0498] R 4 and R 5 All are H,

[0499] R 6 Select one of the following structures:

[0500]

[0501] In one embodiment, the compound has the structure of formula (IIIb),

[0502]

[0503] Among them, -(L) m -Selected from one of the following structures:

[0504]

[0505] R 4 Selected from H, F and C 1-3 Alkoxy, R 5 Selected from H, F and C 1-3 Alkoxy; and R 4 and R 5 There is only one selected from F and C 1-3 Alkoxy, or R 4 and R 5 All are H,

[0506] R 6 Select one of the following structures:

[0507]

[0508] In one embodiment, the compound has the structure of formula (IIIc),

[0509]

[0510] Among them, -(L) m -Selected from one of the following structures:

[0511]

[0512] R 4 and R 5 All are H,

[0513] R 6 Select one of the following structures:

[0514]

[0515] In one embodiment, C 1-3 Alkoxy is selected from methoxy, the halogen is selected from F or Cl, the C 3-6 Cycloalkyl is cyclopropyl, the C 2-6 Alkynyl is ethynyl.

[0516] In one embodiment, R 5 is selected from unsubstituted or optionally substituted with 1 or 2 R 51A phenyl and a 5-6 membered heteroaryl, wherein the 5-6 membered heteroaryl contains 1 or 2 ring-forming heteroatoms selected from N, O or S;

[0517] In one embodiment, R 5 is selected from phenyl, pyridinyl, imidazolyl, pyrazolyl, oxazolyl and thiazolyl; and preferably selected from pyridinyl, imidazolyl and pyrazolyl; said group is unsubstituted or optionally substituted with 1 or 2 R 51A .

[0518] In one embodiment, R 5 is selected from unsubstituted or optionally substituted with 1, 2 or 3 R 51B The 5-7 membered monocyclic alicyclic group or the 7-10 membered bicyclic alicyclic group may be a bridged ring or a spiro ring.

[0519] In one embodiment, R 5 is selected from unsubstituted or optionally substituted with 1, 2 or 3 R 51B The 5-7 membered monocyclic alicyclic group is preferably a 5-7 membered saturated monocyclic alicyclic group, such as tetrahydropyrrolyl, tetrahydrofuranyl, piperidinyl, tetrahydropyranyl, piperazinyl and morpholinyl.

[0520] In one embodiment, R 5 is selected from unsubstituted or optionally substituted with 1, 2 or 3 R 51B For example, the monocyclic alicyclic group is selected from one of the following ring systems:

[0521]

[0522] In one embodiment, R 5 is selected from unsubstituted or optionally substituted with 1, 2 or 3 R 51B For example, the bicyclic alicyclic group is selected from one of the following ring systems:

[0523]

[0524] In one embodiment, R 51A Independently selected from F, Cl, -OR a1 、-NR a1 R b1 、-NR a1 C(=O)R b1 、-NR e1 C(=O)NR a1 R b1 、-NR a1 S(=O)2R d1 、-NR e1 S(=O)2NR a1 R b1 、-C(=O)R a1 、-C(=O)OR a1 、-C(=O)NR a1 R b1 and cyano, and unsubstituted or optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl;

[0525] In one embodiment, R 51A independently selected from unsubstituted or optionally substituted with 1, 2 or 3 R 52 A 5-6 membered aliphatic heterocyclic group.

[0526] In one embodiment, R 51A Selected from:

[0527] 1)F, Cl, -OR a1 、-NR a1 R b1 、-NR a1 C(=O)R b1 、-NRa1 S(=O)2R d1 、-C(=O)NR a1 R b1 、C 1-3 Alkyl, C 1-3 Fluorinated alkyl, C 1-3 Hydroxyalkyl, C 1-3 Cyanoalkyl, cyclopropyl and cyano,

[0528] 2) a 5-6 membered aliphatic heterocyclic group, wherein the 5-6 membered aliphatic heterocyclic group is selected from one of the following structures:

[0529]

[0530] The above structures are unsubstituted or optionally substituted with 1, 2 or 3 R 52 .

[0531] In one embodiment, R 51A Selected from F, Cl, -OR a1 、-NR a1 R b1 、-NR a1 C(=O)R b1 、-NR a1 S(=O)2R d1 、-C(=O)NR a1 R b1 、C 1-3 Alkyl, C 1-3 Fluorinated alkyl, C 1-3 Hydroxyalkyl, C 1-3 Cyanoalkyl, cyclopropyl and cyano.

[0532] In one embodiment, R 51A Independently selected from F, Cl, methoxy, difluoromethoxy, trifluoromethoxy, cyclopropyloxy, methylamino, dimethylamino, cyclopropylamino, cyano, methyl, difluoromethyl, trifluoromethyl and cyclopropyl.

[0533] In one embodiment, R 51A Independently selected from F, Cl, methoxy, methyl, cyclopropyl and cyano.

[0534] In one embodiment, R 51A Independently selected from one of the following structures:

[0535]

[0536] The above structures are unsubstituted or optionally substituted with 1, 2 or 3 R 52 ;

[0537] When R 51A Selected from When R 52 Selected from oxo, methyl, difluoromethyl, trifluoromethyl, cyclopropyl,

[0538] When R 51A When other structures are selected, R 52 Selected from oxo, F, hydroxy, methoxy, trifluoromethoxy, methyl, difluoromethyl, trifluoromethyl, cyclopropyl, cyano,

[0539] In one embodiment, R 5 One of the following ring systems:

[0540]

[0541] The above structures are unsubstituted or optionally substituted with 1, 2 or 3 R 51B , where R 51B Selected from:

[0542] 1) Oxo, F, hydroxy, methoxy, trifluoromethoxy, methylamino, dimethylamino, cyclopropylamino, methyl, difluoromethyl, trifluoromethyl, isopropyl, cyclopropyl, cyano,

[0543] 2) Two R attached to the same carbon atom 51B Together with the carbon atom,

[0544] In one embodiment, R 5 One of the following ring systems:

[0545]

[0546] The above structure is unsubstituted or optionally substituted with a 4-6 membered saturated alicyclic group, wherein the 4-6 membered saturated alicyclic group is unsubstituted or optionally substituted with 1, 2 or 3 substituents selected from F, hydroxyl or methyl, and the 4-6 membered saturated alicyclic group is selected from one of the following ring systems:

[0547]

[0548] In one embodiment, R 51B Selected from:

[0549] 1) F, hydroxy, methyl, isopropyl and cyano,

[0550] 2) Two R attached to the same carbon atom 51B Together with the carbon atom,

[0551] In one embodiment, R 5 Select one of the following structures:

[0552]

[0553] In one embodiment, R 6 Select one of the following structures:

[0554] in,

[0555] R A is selected from H, C optionally substituted with 1, 2 or 3 fluorine 1-3 Alkyl, C 1-3 Hydroxyalkyl, C 1-3 Cyanoalkyl,

[0556] R B Selected from H, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino, optionally substituted with 1, 2 or 3 fluorine C 1-3 Alkyl, C 1-3 Hydroxyalkyl, cyclopropyl, cyano,

[0557] described The ring carbon atoms in the ring are unsubstituted or optionally substituted with 1, 2 or 3 C 1-3 Alkyl The ring carbon atoms in the ring are unsubstituted or optionally substituted with 1, 2 or 3 R G Substitution; and, two R attached to the same carbon atom B Different from H, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino,

[0558] E 2 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(R Y )-、 where R Y Selected from isopropyl,

[0559] E 4 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(CH2CN)-,

[0560] E 5 Selected from -CH2-, -C(CH3)2-, -CF2-, where R Z Selected from H and C 1-3 alkyl;

[0561] The E ring is selected from one of the following structures:

[0562]

[0563] The ring carbon atoms are unsubstituted or optionally replaced by 1, 2 or 3 C 1-3 Alkyl substitution,

[0564] q is selected from 1 and 2.

[0565] In one embodiment, R 6 Select one of the following structures:

[0566] in,

[0567] R A is selected from methyl, isopropyl, C optionally substituted with 1, 2 or 3 fluorine 1-3 Alkyl, C 1-3 Hydroxyalkyl,

[0568] R B Selected from H, hydroxy, methoxy, dimethylamino, methyl, isopropyl, C 1-3 Hydroxyalkyl, described The ring carbon atoms in are unsubstituted or optionally substituted with 1, 2 or 3 substituents selected from F, hydroxyl or methyl; and the two R attached to the same carbon atom B Not at the same time H, hydroxyl, methoxy, dimethylamino,

[0569] E 2 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(R Y )-and where R Y Selected from isopropyl,

[0570] E 4 Selected from carbonyl, -CH2-, -C(CH3)2- and

[0571] The E ring is selected from one of the following structures:

[0572]

[0573] The ring carbon atoms are unsubstituted,

[0574] q is selected from 1 and 2.

[0575] In one embodiment, R 6 Select one of the following structures:

[0576] in,

[0577] R A is selected from methyl, optionally substituted with 1, 2 or 3 fluorine-substituted C 1-3 Alkyl and C 1-3 hydroxyalkyl;

[0578] R B Selected from H, hydroxyl, C 1-3 Hydroxyalkyl, described The ring carbon atoms in the ring are unsubstituted or optionally substituted with 1, 2 or 3 substituents selected from hydroxyl or methyl; and the two R B Different from H, hydroxyl,

[0579] E 4 is selected from carbonyl and -CH2-;

[0580] The E ring is selected from one of the following structures:

[0581] The ring carbon atoms are unsubstituted,

[0582] q is selected from 1 and 2.

[0583] In one embodiment, R 6 Select one of the following structures:

[0584]

[0585] In one embodiment, R 6 Select one of the following structures:

[0586] in,

[0587] R A Selected from H, optionally substituted with 1, 2 or 3 R A11 C 1-3 alkyl, Among them, R A11is selected from fluoro, hydroxy or cyano;

[0588] R B Selected from H, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino, optionally substituted with 1, 2 or 3 R B11 C 1-3 Alkyl, cyclopropyl, cyano, Among them, R B11 Selected from fluorine or hydroxyl; The ring carbon atoms in the ring are unsubstituted or optionally substituted with 1, 2 or 3 C 1-3 Alkyl, the The ring carbon atoms in are unsubstituted or optionally substituted with 1, 2 or 3 R G ; and, the two R linked to the same carbon atom B Different from H, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino,

[0589] E 1 Selected from -CH(R x )-、 where R X Selected from

[0590] E 3 Selected from -C(CH3)2-, -CH(CH2CN)-,

[0591] E 5 Selected from -CH2-, -C(CH3)2-, -CF2-, where R Z Selected from H and C 1-3 alkyl;

[0592] The E ring is a 3-membered saturated alicyclic ring or a 4-6-membered saturated alicyclic heterocyclic ring, wherein the saturated alicyclic heterocyclic ring contains 1 or 2 ring-forming heteroatoms selected from N and O, and the ring-forming carbon atoms of the saturated alicyclic heterocyclic ring are unsubstituted or optionally replaced by 1, 2 or 3 C 1-3 Alkyl substitution, saturated aliphatic heterocyclic ring nitrogen atoms (if any) are optionally replaced by R A replace;

[0593] q is selected from 1 and 2.

[0594] In one embodiment, R 6 Select one of the following structures:

[0595] in,

[0596] R A Selected from H, optionally substituted with 1, 2 or 3 R A11 C 1-3 alkyl, Among them, R A11 is selected from fluoro, hydroxy or cyano;

[0597] R B Selected from H, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino, optionally substituted with 1, 2 or 3 R B11 C 1-3 Alkyl, cyclopropyl, cyano, Among them, R B11 Selected from fluorine or hydroxyl; The ring carbon atoms in the ring are unsubstituted or optionally substituted with 1, 2 or 3 C 1-3 Alkyl, the The ring carbon atoms in the ring are unsubstituted or optionally substituted with 1, 2 or 3 R G ; and, the two R linked to the same carbon atom B Different from H, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino,

[0598] The E ring is selected from one of the following structures:

[0599]

[0600] The ring carbon atoms are unsubstituted or optionally substituted with 1, 2 or 3 C 1-3 alkyl;

[0601] q is selected from 1 and 2.

[0602] In one embodiment, R 6 Select one of the following structures:

[0603] in,

[0604] R B Selected from H, hydroxyl, C 1-3 Hydroxyalkyl, described The ring carbon atoms in the ring are unsubstituted or optionally substituted with 1, 2 or 3 hydroxyl or methyl groups; and the two R BDifferent from H, hydroxyl,

[0605] The E ring is selected from one of the following structures:

[0606] wherein the ring-forming carbon atoms are unsubstituted;

[0607] q is selected from 1 and 2.

[0608] In one embodiment, R 6 Select one of the following structures:

[0609]

[0610] In one embodiment, preferably, R 6 Select one of the following structures:

[0611] or

[0612] In one embodiment, m is selected from 4 and 5, and is preferably 4.

[0613] In one embodiment, n is selected from 0 and 1.

[0614] In one embodiment, n is selected from 1.

[0615] In one embodiment, n is selected from 0.

[0616] In this application, the compounds do not include the following compounds:

[0617]

[0618]

[0619]

[0620]

[0621] In one embodiment, the compound of formula (I) is selected from the following structures:

[0622]

[0623]

[0624]

[0625]

[0626]

[0627]

[0628] synthesis

[0629] The compounds of the present invention and their salts can be prepared using known organic synthesis techniques and can be prepared according to any of a number of possible synthetic routes, such as those in the schemes below.

[0630] The reaction for preparing the compounds of this invention can be implemented in a suitable solvent. Suitable solvents can be substantially unreactive with starting material (reactant), intermediate or product at a temperature (e.g., a temperature within the range of the solvent's freezing temperature to the solvent's boiling temperature) in which the reaction is carried out. A given reaction can be implemented in a mixture of a solvent or multiple solvents. According to a specific reaction step, one skilled in the art can select a suitable solvent for the specific reaction step.

[0631] The preparation of the compounds of the present invention may involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the selection of appropriate protecting groups can be readily determined by one skilled in the art.

[0632] The following schemes provide general guidance for preparing compounds of the present invention. One skilled in the art will appreciate that general knowledge of organic chemistry can be used to modify or optimize the methods shown in the schemes to prepare various compounds of the present invention.

[0633] Compounds of formula (I) can be prepared according to the methods illustrated in the following schemes. For example, various compounds of formula (I) can be prepared using the methods illustrated in Scheme 1. In the method shown in Scheme 1, W is preferably CH.

[0634] First, the compound of formula (1-1) and the compound of formula (INT-1A) undergo a transition metal-catalyzed cross-coupling reaction (eg, Suzuki reaction, Stille reaction, Negishi reaction, etc.) to form the compound of formula (1-2A).

[0635] The compound of formula (1-2) and the compound of formula (INT-2A) are coupled by transition metal catalysis

[0636] reacting to form a compound of formula (1-3A);

[0637] Alternatively, in some embodiments, the compound of formula (1-2) is reacted with the compound of formula (INT-2B) via a transition metal-catalyzed coupling reaction to form the compound of formula (1-3B).

[0638] Among them, X 1 and X 2is halogen (such as chlorine, bromine or iodine) or halogen-like (such as trifluoromethanesulfonyloxy), [M] 1 and [M] 2 It is a metal analogue (such as boric acid, boric ester, fluoroborate, trialkyltin group, organozinc reagent, Grignard reagent, etc.), and the phenolic hydroxyl group and alcoholic hydroxyl group in the compounds of formula (INT-1A), formula (INT-2A) and formula (INT-2B) may not have a protecting group or may have a protecting group (such as acetyl, tetrahydropyran-2-yl, trialkylsilyl, 2-(trimethylsilyl)ethoxy)methyl, etc.).

[0639] Alternatively, in some embodiments, a compound of formula (1-1) is reacted with a compound of formula (INT-1B) via a transition metal-catalyzed coupling reaction to form a compound of formula (1-2B). A compound of formula (1-2B) is reacted with a compound of formula (INT-2B) via a transition metal-catalyzed coupling reaction to form a compound of formula (1-3C).

[0640] In some embodiments, the compound of formula (1-2A) can also be formed into the compound of formula (1-2B) through a substitution reaction; wherein, Y 1 is a good leaving group (selected from bromine and iodine, or from halogen-like groups such as various sulfonates).

[0641] The compounds of formula (1-3A), formula (1-3B) and formula (1-3C) are subjected to Mitsunobu reaction to form a compound of formula (1-4), wherein Y 1 and Y 2 is a good leaving group (selected from, for example, bromine and iodine, or from halogen-like groups, such as various sulfonates); if the alcoholic hydroxyl group and / or phenolic hydroxyl group of the compounds of formula (1-3A), formula (1-3B) and formula (1-3C) carries a protecting group, the protecting group should be removed at an appropriate step before the ring-closure reaction;

[0642] In some embodiments, the compound of formula (1-3A) can be sequentially subjected to halogenation reaction and substitution reaction to form a compound of formula (1-4); wherein the halogen in the halogenation reaction can be selected from bromine and iodine, or selected from halogen-like elements such as various sulfonates; wherein, when L directly linked to the B ring is selected from -O-, m is preferably 5 and 6.

[0643] In some embodiments, the compound of formula (1-3C) can also undergo a substitution reaction to form a compound of formula (1-4), wherein (L) m-1 The terminal group can be a good leaving group or can be formed into a good leaving group through a simple functional group transformation.

[0644] The compound of formula (1-4) is deprotected to form the target compound of formula (I).

[0645] Option 1:

[0646]

[0647] Alternatively, various intermediate compounds of the compounds of formula (I) can be prepared using the method as illustrated in Scheme 2. In the method shown in Scheme 2, W is preferably CH.

[0648] First, the compound of formula (2-1) and the compound of formula (INT-2A) undergo a transition metal-catalyzed coupling reaction to form the compound of formula (2-2A). The compound of formula (2-2A) directly undergoes a halogenation reaction to form the compound of formula (2-3A), wherein X 1 Selected from chlorine, bromine or iodine.

[0649] In some embodiments, the compound of formula (2-2A) can be first subjected to a deprotection reaction to remove the protecting group PG 1 , after halogenation reaction, the protective group PG is introduced to form formula (2-3A)

[0650] Compound; wherein the protective group PG and PG 1 The groups (e.g., tetrahydropyran-2-yl, 2-(trimethylsilyl)ethoxy)methyl, trityl, p-methoxybenzyl, etc.) may be the same or different.

[0651] The compound of formula (2-3A) and the compound of formula (INT-1A) undergo a transition metal-catalyzed coupling reaction to form the compound of formula (1-3A).

[0652] or,

[0653] The compound of formula (2-1) and the compound of formula (INT-2B) undergo a transition metal-catalyzed coupling reaction to form the compound of formula (2-2B). The compound of formula (2-2B) directly undergoes a halogenation reaction to form the compound of formula (2-3B).

[0654] In some embodiments, the compound of formula (2-2B) can be first subjected to a deprotection reaction to remove the protecting group PG 1 , after halogenation reaction, the protective group PG is introduced to form formula (2-3B)

[0655] Compound; wherein the protective group PG and PG 1 It can be the same or different.

[0656] The compound of formula (2-3B) and the compound of formula (INT-1A) undergo a transition metal-catalyzed coupling reaction to form a compound of formula (1-3B);

[0657] In some embodiments, a compound of formula (2-3B) is reacted with a compound of formula (INT-1B) via a transition metal-catalyzed coupling reaction to form a compound of formula (1-3C);

[0658] Wherein, the phenolic hydroxyl group of the compound of formula (2-3B) carries a protective group PG 2 (selected from, for example, tetrahydropyran-2-yl, 2-(trimethylsilyl)ethoxy)methyl and trialkylsilyl, when PG 2 When PG is H, there is no protecting group. 2 When it is not H, the protecting group is removed at an appropriate step before the ring closure reaction.

[0659] The compounds of formula (1-3A), formula (1-3B) and formula (1-3C) are converted into the target compound of formula (I) through the corresponding method of scheme 1.

[0660] Option 2:

[0661]

[0662] In some embodiments, R in the compound of formula (1-4) 5 or R 6 The group is selected from Boc protected amino groups. The compound of formula (1-4) is deprotected to form the compound of formula (1-5). The compound of formula (1-5) is subjected to Sandmeyer reaction to form the compound of formula (1-6); wherein X 3 is selected from halogen, and is preferably bromine and iodine. The compound of formula (1-6) undergoes a transition metal-catalyzed coupling reaction (such as Suzuki reaction, Buchwald-Hartwig amination, etc.) to form the compound of formula (1-7). The compound of formula (1-7) undergoes a deprotection reaction to form the target compound of formula (I). As shown below,

[0663]

[0664] In some embodiments, R in the compound of formula (1-4) 5 or R 6 The group is selected from X 3 ; where X 3 The compound of formula (1-4) is subjected to a transition metal-catalyzed coupling reaction to form a compound of formula (1-7). The compound of formula (1-7) is subjected to a deprotection reaction to form the target compound of formula (I). As shown below,

[0665]

[0666] In some embodiments, X=C-CN, Y=C, Z=CH, there is a double bond between X and Y, and the compound of formula (I) has the structure of formula (II-3),

[0667]

[0668] Wherein, W is preferably CH. L directly linked to ring B is selected from -O-. The compound of formula (3-1) is subjected to cyano reduction reaction and / or other functional group conversion reaction to form a compound of formula (3-2); wherein the group E is selected from aldehyde group, acetal group, formaldehyde oxime group or O-substituted formaldehyde oxime group, and X is 4 Selected from H or halogen. The compound of formula (3-2) undergoes a series of reactions as described in Scheme 1 or Scheme 2 to form the compound of formula (3-3A) or (3-3B). The compound of formula (3-3A) or (3-3B) undergoes a ring closure reaction as described in Scheme 1 or Scheme 2 to form the compound of formula (3-4). The compound of formula (3-4) undergoes a functional group conversion reaction to form the compound of formula (3-5). The compound of formula (3-5) undergoes deprotection to form the target compound of formula (IIb).

[0669] As shown below,

[0670]

[0671] In some embodiments, X=carbonyl, Y=Z=N, there is a single bond between X and Y, and the compound of formula (I) has the structure of formula (II-4),

[0672]

[0673] Wherein, W is preferably CH. The compounds of formula (2-3A) and formula (2-3B) have the structure of formula (4-8) and can be formed by the following method:

[0674]

[0675] First, the compound of formula (4-1) undergoes diazotization and coupling reaction to form the compound of formula (4-2). The compound of formula (4-2) undergoes a ring-closure reaction (e.g., thermal ring-closure, or acid- or base-catalyzed ring-closure reaction) to form the compound of formula (4-3). The compound of formula (4-3) undergoes a chlorination reaction to form the compound of formula (4-4). The compound of formula (4-4) and hydrazine (or hydrazine hydrate) undergo a condensation reaction to form the compound of formula (4-5), wherein Q is selected from an alkoxy group or a hydrazine group. The compound of formula (4-5) undergoes a ring-closure reaction (e.g., thermal ring-closure, or acid- or base-catalyzed ring-closure reaction) to form the compound of formula (4-6). The compound of formula (4-6) undergoes a substitution reaction (e.g., chlorination, bromination, etc.) to form the compound of formula (4-7), wherein X is selected from chlorine, bromine, or trifluoromethanesulfonyloxy. The NH group of the pyrazole is protected with a suitable protecting group to form the compound of formula (4-8). The compound of formula (4-8) is reacted as described in Scheme 2 to form the target compound of formula (II-4).

[0676] In some embodiments, X = carbonyl, Y = Z = N, there is a single bond between X and Y, and the compound of formula (I) has a structure of formula (II-4), wherein W is preferably CH. The compounds of formula (1-3A), formula (1-3B), and formula (1-3C) have structures such as formula (5-8) and can be formed by the following method:

[0677]

[0678] First, the aldehyde compound of formula (5-1) undergoes addition reaction and oxidation reaction (such as using Dess-Martin oxidant) to form the compound of formula (5-3). The compound of formula (4-1) is diazotized and coupled with the compound of formula (5-3) to form the compound of formula (5-4). The compound of formula (5-4) undergoes ring closure reaction to form the compound of formula (5-5). The compound of formula (5-5) is condensed with hydrazine (or hydrazine hydrate) to form the compound of formula (5-6). The compound of formula (5-6) is protected by selecting a suitable protecting group for the NH group of pyrazole to form the compound of formula (5-7). The PG of the phenolic hydroxyl group of the compound of formula (5-7) is removed. 3 The protecting group forms a compound of formula (5-8). The compound of formula (5-8) undergoes the corresponding reaction as in Scheme 1 to form the target compound of formula (II-4).

[0679] Unless otherwise specified or clearly contradicted by the context, all methods described in this specification can be performed in any suitable order. The use of any and all examples or exemplary language (e.g., "such as") provided in this specification is only used to better illustrate the present invention and does not limit the scope of the present invention otherwise claimed.

[0680] Composition and application

[0681] One embodiment of the present invention provides a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, and a pharmaceutically acceptable carrier.

[0682] The pharmaceutical composition can be prepared in a manner well known in the pharmaceutical field and can be administered by various routes. The administration route can be topical (including transdermal, epidermal, ocular, local injection and mucosal, including intranasal, vaginal and rectal delivery), pulmonary (intratracheal or intranasal, such as inhalation or insufflation of powders or aerosols, including by nebulizer), oral or parenteral.

[0683] In one embodiment, the composition is suitable for parenteral administration. This includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular, and other local injections or infusions; or intracranial, such as intrathecal or intraventricular administration. Parenteral administration can be in the form of a single bolus dose or can be administered by, for example, a continuous infusion pump.

[0684] In one embodiment, the composition is suitable for topical administration. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powdered, or oily bases, thickeners, and the like may be necessary or desirable.

[0685] One embodiment of the present invention provides the use of the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and the pharmaceutical composition in preventing or treating diseases mediated by protein kinases.

[0686] One embodiment of the present invention provides the use of the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and the pharmaceutical composition in preventing or treating diseases mediated by HPK1.

[0687] One embodiment of the present invention provides a method for regulating (e.g., inhibiting) the activity of protein kinases such as HPK1, FLT3, KDR, etc., comprising administering to a patient a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof, as well as the above-mentioned pharmaceutical composition, to stimulate and / or enhance the immune response in cancer.

[0688] One embodiment of the present invention provides a method for simultaneously regulating (e.g., inhibiting) the activity of multiple kinase targets (e.g., HPK1, FLT3, KDR, etc.), comprising administering to a patient a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, as well as the pharmaceutical composition described above, to inhibit the growth of tumor cells.

[0689] One embodiment of the present invention provides a method for simultaneously regulating (e.g., inhibiting) the activity of HPK1 and multiple other kinase targets (e.g., FLT3, KDR, etc.), comprising administering to a patient a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope label, isomer or prodrug thereof, as well as the pharmaceutical composition thereof, to stimulate and / or enhance the immune response in cancer and inhibit the growth of tumor cells.

[0690] One embodiment of the present invention provides a method for preventing, ameliorating or treating diseases mediated by protein kinases such as HPK1, FLT3, KDR, etc., comprising administering a therapeutically effective amount of the above-mentioned compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and the above-mentioned pharmaceutical composition to a patient suffering from a disease mediated by protein kinases such as HPK1, FLT3, KDR, etc.

[0691] One embodiment of the present invention provides a method for preventing, ameliorating or treating tumors (including benign and malignant tumors), comprising administering to a patient suffering from a tumor a therapeutically effective amount of the compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, as well as the pharmaceutical composition described above.

[0692] One embodiment of the present invention provides a method for preventing, ameliorating or treating a disease caused by a virus, comprising administering a therapeutically effective amount of the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, as well as the above-mentioned pharmaceutical composition to a patient suffering from a tumor.

[0693] One embodiment of the present invention provides a method for preventing, ameliorating or treating myelodysplastic syndrome, comprising administering a therapeutically effective amount of the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled form, isomer or prodrug, and the pharmaceutical composition thereof to a patient suffering from a tumor.

[0694] One embodiment of the present invention provides the use of the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and the pharmaceutical composition in treating or ameliorating specific diseases, including one or more of tumors, myelodysplastic syndrome and viral-induced diseases.

[0695] One embodiment of the present invention provides the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and the use of the above-mentioned pharmaceutical composition in treating or ameliorating tumors.

[0696] One embodiment of the present invention provides the compound of formula (I) or its pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug, and the use of the above-mentioned pharmaceutical composition in treating or ameliorating viral diseases.

[0697] One embodiment of the present invention provides a use of the compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled form, isomer or prodrug thereof, or the pharmaceutical composition thereof in the preparation of a medicament for treating or ameliorating a specific disease, wherein the disease comprises one or more of a tumor, myelodysplastic syndrome and a virus-induced disease.

[0698] One embodiment of the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, or the use of the above-mentioned pharmaceutical composition in the preparation of a drug for treating or ameliorating tumors.

[0699] One embodiment of the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt, hydrate, solvate, active metabolite, polymorph, isotope-labeled substance, isomer or prodrug thereof, or the use of the above-mentioned pharmaceutical composition in the preparation of a medicament for treating or ameliorating a disease caused by a virus.

[0700] In one embodiment, the tumor comprises one or more of chronic or acute leukemia, lymphoma, primary CNS lymphoma, multiple myeloma, lung cancer, hepatocellular carcinoma, bile duct cancer, gallbladder cancer, gastric cancer, colorectal cancer, small intestinal leiomyosarcoma, breast cancer, triple-negative breast cancer, ovarian cancer, cervical cancer, endometrial cancer, fallopian tube cancer, vaginal cancer, vulvar cancer, malignant teratoma, pancreatic cancer, pancreatic ductal adenocarcinoma, nasopharyngeal cancer, oral cancer, laryngeal cancer, hypopharyngeal cancer, esophageal squamous cell carcinoma, esophageal adenocarcinoma, thyroid cancer, kidney cancer, bladder cancer, malignant brain tumor, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, fibrosarcoma of bone, Ewing's sarcoma, myxoma, malignant thymoma, malignant peripheral nerve sheath tumor, prostate cancer, testicular cancer, penile cancer, urethral cancer, and skin malignancies (including squamous cell carcinoma, basal cell carcinoma, malignant melanoma, etc.).

[0701] In one embodiment, the disease-causing virus includes one or more of hepatitis virus, human immunodeficiency virus, human papillomavirus, herpes simplex virus, measles virus, norovirus, bocavirus, coxsackievirus, Ebola virus, enterovirus, lymphocytic meningitis virus, influenza virus, SARS virus and new coronavirus.

[0702] The preparation and properties of the compound of formula (I) according to one embodiment of the present invention are further described below with reference to specific examples. The starting materials used are known and commercially available, or can be synthesized using or according to methods known in the art.

[0703] Unless otherwise specified, all reactions in the examples were carried out under continuous magnetic stirring, and the reaction temperatures were expressed in degrees Celsius.

[0704] The reaction can be monitored by any suitable method known in the art, such as nuclear magnetic resonance spectroscopy (NMR), infrared spectroscopy (IR), spectrophotometry (such as UV-visible spectroscopy), liquid chromatography-mass spectrometry (LC-MS), mass spectrometry, high performance liquid chromatography, thin layer chromatography (or thin layer chromatography), etc. The product can be purified by any suitable method known in the art, such as column chromatography (normal phase or reverse phase), preparative thin layer chromatography, beating, recrystallization, etc. Normal phase column chromatography usually uses Qingdao Ocean 100-200 mesh silica gel as a carrier. Thin layer chromatography (TLC) uses Merck's Silica gel 60F254 silica gel plates, and preparative thin layer chromatography (pre-TLC) uses Anhui Liangchen Silicon Source GF254 preparative silica gel plates.

[0705] The structures of the compounds of the present invention are determined by nuclear magnetic resonance spectroscopy (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). The NMR spectrum is measured by a Bruker AVANCE-400 nuclear magnetic resonance instrument, and the measuring solvent is usually deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDCl3). The NMR chemical shift (δ) is given in parts per million (ppm), and the internal standard is tetramethylsilane (TMS). The LC-MS is measured by an Agilent 1100 series liquid chromatograph and a Bruker HCT-Ultra ion trap mass spectrometer.

[0706] Abbreviations

[0707]

[0708]

[0709] Synthesis Example: Preparation of 3-fluoro-2-(3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-5-yl)phenol (Intermediate A1)

[0710]

[0711] Step 1: Preparation of tert-butyl 5-bromo-1H-pyrazolo[3,4-c]pyridine-1-carboxylate (Intermediate A1-A1)

[0712]

[0713] Under nitrogen, 5-bromo-1H-pyrazolo[3,4-c]pyridine (10.0 g, 50.5 mmol) was dissolved in 1,4-dioxane (100 mL), and DIPEA (9.8 g, 75.8 mmol) and DMAP (62 mg, 76 mmol) were added. Di-tert-butyl dicarbonate (12.12 g, 55.60 mmol) was added at 0°C. The reaction was allowed to proceed at room temperature for 18 h. The mixture was quenched with water (100 mL), extracted with ethyl acetate (150 mL × 3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (petroleum ether / ethyl acetate = 5 / 1) to give intermediate A1-A1 (14.1 g, white solid) in a yield of 94%.

[0714] ESI-MS: m / z = 298.2, [M+H] + .

[0715] Step 2: Preparation of (2-((2-bromo-3-fluorophenoxy)methoxy)ethyl)trimethylsilane (Intermediate A1-A2)

[0716]

[0717] At room temperature, anhydrous potassium carbonate (32.513 g, 235.60 mmol) was added to a solution of 2-bromo-3-fluorophenol (15.000 g, 78.53 mmol) in DMF (150 mL), followed by the dropwise addition of 2-(trimethylsilyl)ethoxymethyl chloride (15.711 g, 94.24 mmol), and the mixture was stirred at room temperature for 1 hour. Water (600 mL) was added to the reaction system, which was then extracted with ethyl acetate (400 mL × 2). The organic phases were combined and washed once with water (300 mL) and saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to give intermediate A1-A2 (18.10 g, colorless oil) in a 77% yield.

[0718] Step 3: Preparation of 2-fluoro-6-((2-(trimethylsilyl)ethoxy)methoxy)phenylboronic acid pinacol ester (Intermediate A1-A3)

[0719]

[0720] Under nitrogen protection and a dry ice-ethanol bath, a solution of intermediate A1-A2 (13.000 g, 40.47 mmol) in dry tetrahydrofuran (130 mL) was cooled to -70°C. A 2.5 M solution of n-butyllithium in hexane (19.4 mL, 48.6 mmol) was then slowly added dropwise, maintaining the internal temperature below -60°C. After the addition was complete, the mixture was stirred at -65°C for 1 hour. Isopropyl alcohol pinacol borate (11.293 g, 60.67 mmol) was then slowly added dropwise, and the mixture was stirred at -65°C for 1 hour. The reaction was quenched with ammonium chloride solution (130 mL) and extracted with ethyl acetate (100 mL x 2). The organic phase was washed once with water (100 mL) and once with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude intermediate A1-A3 (15.42 g, colorless oil) was obtained with a yield of 103.4%.

[0721] Step 4: Preparation of 5-(2-fluoro-6-((2-(trimethylsilyl)ethoxy)methoxy)phenyl)-1H-pyrazolo[3,4-c]pyridine (Intermediate A1-A4)

[0722]

[0723] Under nitrogen protection, a solution of intermediate A1-A1 (5.510 g, 18.49 mmol), intermediate A1-A3 (10.896 g, 29.58 mmol), potassium phosphate (11.774 g, 55.47 mmol) and Xphos-Pd-G2 (1.163 g, 1.48 mmol) in 1,4-dioxane (65 mL) and water (13 mL) was heated to 80 ° C and stirred for 2 hours. The mixture was cooled to room temperature and a 2M aqueous lithium hydroxide solution (50 mL) was added dropwise to the system and stirred for 4 hours. The pH was adjusted to 7-8 with a 5% aqueous citric acid solution. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (150 mL × 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to afford intermediate A1-A4 (6.07 g, white solid) in a yield of 91.3%.

[0724] ESI-MS: m / z = 360.3, [M+H] + .

[0725] Step 5: Preparation of 5-(2-fluoro-6-((2-(trimethylsilyl)ethoxy)methoxy)phenyl)-3-iodo-1H-pyrazolo[3,4-c]pyridine (Intermediate A1-A5)

[0726]

[0727] To a solution of intermediate A1-A4 (6.070 g, 16.88 mmol) in DMAc (60 mL) at 0°C were added potassium hydroxide (2.842 g, 50.66 mmol) and iodine (4.718 g, 18.57 mmol), followed by stirring at room temperature overnight. The reaction was quenched with aqueous ammonium chloride (150 mL) and extracted with ethyl acetate (150 mL x 2). The organic phase was washed once with aqueous sodium thiosulfate (100 mL), once with water (100 mL), and once with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate (5 / 1 to 2 / 1) to afford intermediate A1-A5 (7.30 g, white solid) in an 89.1% yield.

[0728] ESI-MS: m / z = 486.3, [M+H] + .

[0729] Step 6: Preparation of Intermediate A1

[0730]

[0731] Under nitrogen, to a solution of Intermediate A1-A5 (8.800 g, 18.13 mmol) in 1,4-dioxane (90 mL) were added 3,4-dihydro-2H-pyran (4.569 g, 54.39 mmol) and p-toluenesulfonic acid monohydrate (1.724 g, 9.07 mmol), then the mixture was heated to 100°C and stirred for 3 hours. The mixture was cooled to room temperature, water (200 mL) was added, and the mixture was extracted with ethyl acetate (150 mL x 2). The organic phase was washed once with water (100 mL) and once with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to obtain Intermediate A1 (6.76 g, pale yellow solid) in an 84.9% yield.

[0732] ESI-MS: m / z = 440.3, [M+H] + .

[0733] Alternatively, intermediate A1 can also be prepared by the following method.

[0734] Synthesis Example: Preparation of Intermediate A1

[0735] Step 1: Preparation of tert-butyl 5-(2-fluoro-6-methoxyphenyl)-1H-pyrazolo[3,4-c]pyridine-1-carboxylate (Intermediate A1-B1)

[0736]

[0737] Under nitrogen protection, intermediate A1-A1 (7.000 g, 23.49 mmol), 2-fluoro-6-methoxyphenylboronic acid (5.230 g, 30.76 mmol), potassium phosphate (14.954 g, 70.47 mmol) and XPhos-Pd-G2 (1.728 g, 1.577 mmol) were added to a mixed solvent of 1,4-dioxane / water (110 mL / 15 mL). The reaction was heated to 90°C for 18 h, and the mixture was quenched by adding water (100 mL). The product was extracted with ethyl acetate (150 mL×3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain intermediate A1-B1 (8.912 g, yellow oil) in a yield of 62%.

[0738] ESI-MS: m / z = 344.2, [M+H] + .

[0739] Step 2: Preparation of 5-(2-fluoro-6-methoxyphenyl)-1H-pyrazolo[3,4-c]pyridine (Intermediate A1-B2)

[0740]

[0741] Intermediate A1-B1 (8.912 g, 25.91 mmol) was dissolved in tetrahydrofuran (15 mL), and saturated aqueous lithium hydroxide solution (8 mL) was added. The mixture was heated to 40°C for 3 hours, and the pH was adjusted to 7 with 6N hydrochloric acid. The mixture was extracted with ethyl acetate (100 mL × 3), washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, and concentrated to give intermediate A1-B2 (6.310 g, yellow solid) in a quantitative yield.

[0742] ESI-MS: m / z = 244.1, [M+H] + .

[0743] Step 3: Preparation of 3-iodo-5-(2-fluoro-6-methoxyphenyl)-1H-pyrazolo[3,4-c]pyridine (Intermediate A1-B3)

[0744]

[0745] Intermediate A1-B2 (6.310 g, 25.86 mmol) was dissolved in DMAc (100 mL), and KOH (4.345 g, 77.46 mmol) and iodine (3.941 g, 31.03 mmol) were added sequentially. The mixture was reacted at room temperature for 18 h, and then quenched by addition of aqueous sodium sulfite solution. The pH was adjusted to 7 with 4N hydrochloric acid, and the mixture was extracted with ethyl acetate (100 mL × 3), washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, and concentrated to afford intermediate A1-B3 (8.832 g, yellow solid) in a 93% yield.

[0746] ESI-MS: m / z = 370.1, [M+H] + .

[0747] Step 4: Preparation of 3-fluoro-2-(3-iodo-1H-pyrazolo[3,4-c]pyridin-5-yl)phenol (Intermediate A1-B4)

[0748]

[0749] Intermediate A1-B3 (12.844 g, 34.8 mmol) was dissolved in dichloromethane (150 mL), cooled to 0°C, and boron tribromide (10.2 mL, 104 mmol) was added dropwise. The reaction was allowed to react at room temperature for 18 h. The mixture was cooled to 0°C, and methanol was added dropwise to quench the reaction. The pH was adjusted to 8 with NaHCO3, and the mixture was extracted with dichloromethane (100 mL × 3), washed with saturated brine (300 mL), and dried over anhydrous sodium sulfate. The crude product was concentrated and purified on a silica gel column (petroleum ether / ethyl acetate = 3 / 1) to obtain intermediate A1-B4 (9.512 g, yellow solid) in a yield of 77%.

[0750] ESI-MS: m / z = 356.1, [M+H] + .

[0751] Step 5: Preparation of Intermediate A1

[0752]

[0753] Intermediate A1-B4 (8.212 g, 23.2 mmol) was dissolved in 1,4-dioxane (100 mL), and 3,4-dihydro-2H-pyran (5.844 g, 69.6 mmol) and p-toluenesulfonic acid monohydrate (2.578 g, 13.54 mmol) were added at room temperature. The mixture was reacted at 85°C for 3 h. Saturated aqueous sodium bicarbonate solution was poured into the system, and the mixture was extracted with dichloromethane (100 mL × 3). The mixture was washed with saturated brine (300 mL) and dried over anhydrous sodium sulfate. The crude product was concentrated and purified on a silica gel column (petroleum ether / ethyl acetate = 10 / 1) to obtain intermediate A1 (10.400 g, yellow solid) with a quantitative yield.

[0754] ESI-MS: m / z = 440.2, [M+H] + .

[0755] Synthesis Example: Preparation of 3-acetoxy-4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenylboronic acid pinacol ester (Intermediate A2)

[0756]

[0757] Step 1: Preparation of (R)-8-(2-(benzyloxy)phenyl)octahydropyrazino[2,1-c][1,4]oxazine (Intermediate A2-1)

[0758]

[0759] To a solution of 2-benzyloxybromobenzene (7152 mg, 27.18 mmol) in toluene (100 mL) were added (R)-octahydropyrazino[2,1-c][1,4]oxazine (5828 mg, 40.98 mmol), palladium acetate (610 mg, 2.72 mmol), cesium carbonate (26620 mg, 81.70 mmol) and BINAP (3411 mg, 5.48 mmol), and the mixture was heated to 120°C and stirred under nitrogen for 14 h. After the reaction, the reaction solution was purified by column chromatography using petroleum ether / ethyl acetate = 5 / 1 to 0 / 1 as the developing solvent to obtain intermediate A2-1 (6137 g, yellow liquid) with a yield of 70%.

[0760] ESI-MS: m / z = 325.2, [M+H] +.

[0761] Step 2: Preparation of (R)-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenol (Intermediate A2-2)

[0762]

[0763] Intermediate A2-1 (6137 mg, 18.92 mmol) was dissolved in methanol (60 mL), and 10% palladium on carbon (606 mg) was added. The hydrogen was replaced and the mixture was stirred at room temperature under a hydrogen atmosphere (1 atm) for 16 hours. After the reaction was completed, the reaction solution was filtered, the filter cake was washed with methanol, the filtrate was collected, and the solvent was removed in vacuo to obtain intermediate A2-2 (4.372 g, white solid) in a yield of 99%.

[0764] ESI-MS: m / z = 235.1, [M+H] + .

[0765] Step 3: Preparation of (R)-(2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenol)acetate (Intermediate A2-3)

[0766]

[0767] Intermediate A2-2 (4372 mg, 18.66 mmol) was dissolved in dichloromethane (50 mL), cooled to 0°C, and pyridine (4.51 mL, 55.98 mmol), acetic anhydride (2.12 mL, 22.4 mmol) and DMAP (30 mg, 0.25 mmol) were added and returned to room temperature. After stirring at room temperature for 3 hours, the reaction was quenched with water (150 mL) and then extracted with ethyl acetate (50 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo to obtain a crude product. The crude product was purified by column chromatography using petroleum ether / ethyl acetate = 2 / 1 to 0 / 1 as eluent to obtain intermediate A2-3 (5191 mg, colorless liquid) in a 100% yield.

[0768] ESI-MS: m / z = 277.2, [M+H] + .

[0769] Step 4: Preparation of (R)-(2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-5-bromophenol)acetate (Intermediate A2-4)

[0770]

[0771] Intermediate A2-3 (1002 mg, 3.63 mmol) was dissolved in dichloromethane (10 mL) and cooled to 0°C. A solution of hydrogen bromide in acetic acid (1.8 mL, 11 mmol) and NBS (966 mg, 5.43 mmol) were then added. After reacting at room temperature for 2 hours, the reaction was quenched with aqueous sodium carbonate (50 mL). The mixture was then extracted with ethyl acetate (50 mL × 3). The organic phase was collected, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo to obtain a crude product. The crude product was purified by column chromatography using petroleum ether / ethyl acetate = 2 / 1 to 0 / 1 as the eluent to obtain intermediate A2-4 (1.149 mg, yellow solid) in an 89% yield.

[0772] ESI-MS: m / z = 355.1, [M+H] + .

[0773] Step 5: Preparation of Intermediate A2

[0774]

[0775] Intermediate A2-4 (1149 mg, 3.23 mmol) was dissolved in 1,4-dioxane (15 mL), and pinacol diboronate (1093 mg, 4.30 mmol), Pd(dppf)Cl2 (237 mg, 0.32 mmol), and potassium acetate (976 mg, 9.95 mmol) were added. The mixture was then heated to 90°C and stirred for 16 hours under nitrogen. After the reaction was completed, the mixture was cooled to room temperature, quenched with water (150 mL), and extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using dichloromethane / methanol = 30 / 1 as the eluent to obtain intermediate A2 (1388 mg, black liquid) in a quantitative yield.

[0776] ESI-MS: m / z = 403.2, [M+H] + .

[0777] Synthesis Example: Preparation of 3-acetoxy-4-(4-(tert-butoxycarbonyl)piperazin-1-yl)phenylboronic acid pinacol ester (Intermediate A3)

[0778]

[0779] Intermediate A3 was prepared by similar procedures to Intermediate A2, using 1-(tert-butoxycarbonyl)piperazine instead of (R)-octahydropyrazino[2,1-c][1,4]oxazine as the starting material.

[0780] Synthesis Example: Preparation of 4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)phenylboronic acid pinacol ester (Intermediate A4)

[0781]

[0782] Step 1: Preparation of (9aR)-8-(2-(3-((tetrahydro-2H-pyran-2-yl)oxy)propyloxy)phenyl)octahydropyrazino[2,1-c][1,4]oxazine (Intermediate A4-1)

[0783]

[0784] Under nitrogen, a toluene (30 mL) solution of 2-(3-(2-bromophenoxy)propoxy)tetrahydro-2H-pyran (2.00 g, 6.35 mmol), (R)-octahydropyrazino[2,1-c][1,4]oxazine (1.173 g, 8.25 mmol), palladium acetate (142 mg, 0.634 mmol), BINAP (790 mg, 1.27 mmol), and cesium carbonate (6.20 g, 19.0 mmol) was heated to 100°C and stirred for 14 hours. The reaction system was cooled to room temperature, water (200 mL) was added, and the mixture was extracted with ethyl acetate (200 mL × 2). The organic phases were combined and washed once with water (200 mL) and once with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 20 / 1 as the eluent to afford intermediate A4-1 (1.67 g, yellow oil) in a 70% yield.

[0785] ESI-MS: m / z = 377.2, [M+H] + .

[0786] Step 2: Preparation of (9aR)-8-(4-bromo-2-(3-((tetrahydro-2H-pyran-2-yl)oxy)propyloxy)phenyl)octahydropyrazino[2,1-c][1,4]oxazine (Intermediate A4-2)

[0787]

[0788] Under nitrogen protection and 0°C, NBS (829 mg, 4.65 mmol) was added in batches to a solution of intermediate A4-1 (1.67 g, 4.43 mmol) in dichloromethane (20 mL), and then the mixture was warmed to room temperature and stirred for 2 hours. The reaction was quenched with aqueous sodium bicarbonate solution and extracted with dichloromethane (100 mL). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography eluting with dichloromethane / methanol = 20 / 1 to give intermediate A4-2 (1.79 g, oily substance) in a yield of 89%.

[0789] ESI-MS: m / z = 455.2, [M+H] + .

[0790] Step 3: Preparation of Intermediate A4

[0791]

[0792] Under nitrogen, a solution of Intermediate A4-2 (1.79 g, 3.95 mmol), bis(pinacol borate) (1.30 g, 5.14 mmol), potassium acetate (1.16 g, 11.80 mmol), and Pd(dppf)Cl2 (288 g, 0.39 mmol) in 1,4-dioxane (25 mL) was heated to 90°C and stirred for 6 hours. The reaction mixture was cooled to room temperature, and most of the solvent was evaporated off. Water (100 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (110 mL x 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography eluting with ethyl acetate to obtain Intermediate A4 (1.70 g, light yellow oil) in an 86% yield.

[0793] ESI-MS: m / z = 503.3, [M+H] + .

[0794] Synthesis Example: Preparation of 4-(4-(tert-Butoxycarbonyl)piperazin-1-yl)-3-(3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)phenylboronic acid pinacol ester (Intermediate A5)

[0795]

[0796] Intermediate A5 was prepared by similar procedures to Intermediate A4, using 1-(tert-butoxycarbonyl)piperazine instead of (R)-octahydropyrazino[2,1-c][1,4]oxazine as the starting material.

[0797] ESI-MS: m / z = 547.3, [M+H] + .

[0798] Example 1

[0799] Compound 1:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 5 -Chloro-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane

[0800]

[0801] Step 1: Preparation of tert-butyl (4-bromo-2-((tert-butoxycarbonyl)oxy)phenyl)carbamate (Intermediate 1-1)

[0802]

[0803] To a solution of 2-amino-5-bromophenol (9173 mg, 48.79 mmol) in dichloromethane (150 mL) was added di-tert-butyl dicarbonate (21.275 g, 97.48 mmol) and stirred at room temperature for 14 h. After the reaction, the reaction solution was purified by column chromatography using petroleum ether / ethyl acetate = 20 / 1 to 10 / 1 as the developing solvent to obtain intermediate 1-1 (12.300 g, white solid) with a yield of 65%.

[0804] ESI-MS: m / z = 388.1, [M+H] + .

[0805] Step 2: Preparation of 4-((tert-butoxycarbonyl)amino)-3-((tert-butoxycarbonyl)oxy)phenylboronic acid pinacol ester (Intermediate 1-2)

[0806]

[0807] Intermediate 1-1 (12.300 g, 31.68 mmol) was dissolved in ultra-dry 1,4-dioxane (120 mL), and diboronic acid pinacol ester (10.480 g, 41.27 mmol), Pd(dppf)Cl2 (2.356 g, 3.22 mmol) and potassium acetate (9.376 g, 95.54 mmol) were added. The mixture was then heated to 100 ° C and stirred for 3 hours under nitrogen protection. After the reaction was completed, it was cooled to room temperature, quenched with water (500 mL), and extracted with ethyl acetate (300 mL × 3). The organic phase was washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 20 / 1 to 10 / 1 to obtain intermediate 1-2 (12.28 g, yellow liquid) with a yield of 89%.

[0808] ESI-MS: m / z = 436.2, [M+H] + .

[0809] Step 3: Preparation of tert-butyl (4-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-hydroxyphenyl)carbamate (Intermediate 1-3)

[0810]

[0811] Intermediate 1-2 (5770 mg, 13.25 mmol) was dissolved in dioxane (90 mL), and Pd(dppf)Cl2 (935 mg, 1.28 mmol), potassium phosphate (8040 mg, 37.88 mmol), 5-bromo-3-iodo-1-(2-(trimethylsilyl)ethoxy)methyl-1H-pyrazolo[3,4-c]pyridine (5730 mg, 12.62 mmol), and water (22.5 mL) were added. The mixture was then protected by nitrogen and stirred at 85°C for 14 hours. After completion of the reaction, water (300 mL) was added to quench the reaction, followed by extraction with ethyl acetate (200 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo to obtain the crude product. The crude product was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to 5 / 1 as the eluent to obtain Intermediate 1-3 (3945 mg, brown liquid) in a yield of 58%.

[0812] ESI-MS: m / z = 535.1, [M+H] + .

[0813] Step 4: Preparation of 2-bromo-4-chloro-3-fluorobenzoic acid (Intermediate 1-4)

[0814]

[0815] Ultra-dry tetrahydrofuran (50 mL) was protected by nitrogen and cooled to -70 ° C. Then, a 2M solution of lithium diisopropylamine in tetrahydrofuran (30.3 mL, 60.5 mmol) was slowly added. 4-Chloro-3-fluorobenzoic acid (4.814 g, 27.6 mmol) was dissolved in ultra-dry tetrahydrofuran (20 mL) and slowly added dropwise to the above system. After the addition was completed, the reaction was kept at low temperature for one hour. Then, 1,2-dibromotetrachloroethane (17937 mg, 55 08mmol) in ultra-dry tetrahydrofuran (50mL) was added dropwise to the reaction system. After the addition was completed, the temperature was slowly raised to room temperature. After reaction at room temperature for 14 hours, ethyl acetate (200mL) was added for dilution, followed by extraction with water (100mL×3). The aqueous phase was collected and adjusted to pH=1-2 with 4M hydrochloric acid, followed by extraction with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo to obtain intermediate 1-4 (6.848g, white solid) with a yield of 98%.

[0816] Step 5: Preparation of 2-bromo-4-chloro-3-fluorobenzyl alcohol (Intermediate 1-5)

[0817]

[0818] Intermediate 1-4 (6548 mg, 25.84 mmol) was dissolved in tetrahydrofuran (70 mL), and borane dimethyl sulfide complex (7.75 mL, 77.5 mmol) was added. The temperature was then raised to 60°C and stirred for 3 hours. After the reaction was completed, the mixture was cooled to 0°C, quenched with methanol (300 mL), and dried in vacuo. Aqueous sodium bicarbonate solution (150 mL) was added, and the mixture was extracted with ethyl acetate (100 mL×3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a crude intermediate 1-5 (6.309 g, white solid).

[0819] Step 6: Preparation of 2-bromo-1-(bromomethyl)-4-chloro-3-fluorobenzene (Intermediate 1-6)

[0820]

[0821] A solution of intermediate 1-5 (6109 mg, 25.51 mmol) in dichloromethane (65 mL) was cooled to 0°C, and then phosphorus tribromide (2.66 mL, 28.1 mmol) was added dropwise. The reaction system was stirred at 0°C for 3 hours. After the reaction was completed, the reaction solution was poured into an aqueous solution of sodium bicarbonate (200 mL, pH = 8), and then extracted with methyl tert-butyl ether (100 mL × 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the product intermediate 1-6 (4937 mg, yellow liquid) with a yield of 63%.

[0822] Step 7: Preparation of 3-((2-bromo-4-chloro-3-fluorobenzyl)oxy)propan-1-ol (Intermediate 1-7)

[0823]

[0824] Ultra-dry tetrahydrofuran (50 mL) was cooled to 0 ° C, and then sodium hydroxide (718 mg, 18.0 mmol) and 1,3-propylene glycol (22.5 mL, 310 mmol) were slowly added in sequence. The nitrogen atmosphere was replaced and the reaction was carried out at 0 ° C for 40 minutes. Then, a solution of intermediate 1-6 (3437 mg, 11.37 mmol) in tetrahydrofuran (10 mL) was added. After the addition was complete, the temperature was slowly raised to 40 ° C. The reaction was allowed to proceed for 14 hours. After the reaction was completed, the temperature was lowered to 0 ° C, water (150 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (100 mL). The organic phase was washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain a crude intermediate 1-7 (3767 mg, yellow liquid). The crude product was used directly in the next step.

[0825] ESI-MS: m / z = 299.0, [M+H] + .

[0826] Step 8: Preparation of 2-(3-((2-bromo-4-chloro-3-fluorobenzyl)oxy)propoxy)tetrahydro-2H-pyran (Intermediate 1-8)

[0827]

[0828] Intermediate 1-7 (3767 mg, 12.66 mmol) was dissolved in dichloromethane (35 mL), and 3,4-dihydro-2H-pyran (1294 mg, 15.40 mmol) and 4-methylbenzenesulfonate pyridinium (641 mg, 2.55 mmol) were added. The mixture was heated to 35°C and reacted for 2 hours. After completion of the reaction, water (150 mL) was added to quench the reaction. The mixture was then extracted with dichloromethane (100 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo to obtain the crude product. The crude product was purified by column chromatography using petroleum ether / ethyl acetate = 20 / 1 as the eluent to obtain Intermediate 1-8 (3799 mg, colorless liquid) with a two-step yield of 88%.

[0829] ESI-MS: m / z = 383.0, [M+H] + .

[0830] Step 9: Preparation of 3-chloro-2-fluoro-6-((3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)methyl)phenylboronic acid pinacol ester (Intermediate 1-9)

[0831]

[0832] A solution of intermediate 1-8 (3299 mg, 8.64 mmol) in ultra-dry tetrahydrofuran (60 mL) was cooled to -70°C, and then a 2.5 M solution of n-butyl lithium in hexane (4.15 mL, 10.4 mmol) was added dropwise. After incubation for 1 hour, isopropyl alcohol pinacol borate (3278 mg, 17.62 mmol) was added dropwise. After the addition was complete, the temperature was maintained for 1.5 hours. After completion of the reaction, aqueous ammonium chloride solution (50 mL) was added to quench the reaction, the mixture was diluted with water (100 mL), and then extracted with dichloromethane (100 mL × 3). The organic phase was collected, washed with saturated brine (100 mL), and dried over anhydrous sodium sulfate. The solvent was evaporated in vacuo to obtain a crude product of intermediate 1-9 (5130 mg, yellow liquid). The crude product was used directly in the next reaction.

[0833] ESI-MS: m / z = 429.2, [M+H] + .

[0834] Step 10: Preparation of tert-butyl (4-(5-(3-chloro-2-fluoro-6-(3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)methyl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-hydroxyphenyl)carbamate (Intermediate 1-10)

[0835]

[0836] To a mixed solution of intermediate 1-9 (5130 mg, 11.97 mmol) in 1,4-dioxane (40 mL) and water (10 mL) was added intermediate 1-3 (2540 mg, 4.74 mmol), Xphos-Pd-G2 (373 mg, 0.474 mmol) and sodium carbonate (1539 mg, 14.52 mmol), then heated to 80 ° C and stirred for 15 hours. After the reaction was completed, it was quenched with water (150 mL) and cooled to room temperature and extracted with ethyl acetate (100 mL × 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography with petroleum ether / ethyl acetate = 5 / 1 to 3 / 1 as eluent to obtain the product intermediate 1-10 (2008 mg, yellow liquid) with a yield of 58%.

[0837] ESI-MS: m / z = 757.3, [M+H] + .

[0838] Step 11: Preparation of tert-butyl (4-(5-(3-chloro-2-fluoro-6-(3-hydroxypropoxy)methyl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-hydroxyphenyl)carbamate (Intermediate 1-11)

[0839]

[0840] To a solution of intermediate 1-10 (2008 mg, 2.65 mmol) in methanol (30 mL) was added p-toluenesulfonic acid monohydrate (526 mg, 2.77 mmol), and the mixture was heated to 40° C. and stirred for 0.5 hour. After the reaction was completed, the methanol was removed by distillation under reduced pressure. The remaining concentrate was dissolved in water (150 mL) and then extracted with dichloromethane (100 mL×3). The organic phase was collected, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was evaporated in vacuo, and the residue was purified by column chromatography with petroleum ether / ethyl acetate = 3 / 1 to 1 / 1 as the eluent to obtain the product intermediate 1-11 (1457 mg, yellow liquid) in a yield of 81%.

[0841] ESI-MS: m / z = 673.3, [M+H] + .

[0842] Step 12:1 6 -(tert-Butyloxycarbonylamino)-2- ... 1 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 5 -Chloro-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 1-12)

[0843]

[0844] To a solution of TMAD (1745 mg, 10.13 mmol) in ultra-dry tetrahydrofuran (30 mL) was added tri-n-butylphosphine (2071 mg, 10.23 mmol). The mixture was reacted at room temperature under nitrogen for 10 minutes. Ultra-dry tetrahydrofuran (90 mL) was then added to dilute the system. The temperature was raised to 50°C and a solution of intermediate 1-11 (1357 mg, 2.02 mmol) in tetrahydrofuran was added dropwise. The mixture was reacted at 50°C under nitrogen for 20 minutes. After the reaction was completed, water (200 mL) was added to quench the mixture, followed by extraction with ethyl acetate (100 mL × 3). The organic phase was collected, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was evaporated in vacuo, and the residue was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to 5 / 1 as the eluent to obtain intermediate 1-12 (1008 mg, yellow liquid) in a yield of 71%.

[0845] ESI-MS: m / z = 655.2, [M+H] + .

[0846] Step 13:1 6 -Amino-2 1 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 5 -Chloro-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 1-13)

[0847]

[0848] To a solution of intermediate 1-12 (967 mg, 1.47 mmol) in methanol (20 mL) was added p-toluenesulfonic acid monohydrate (703 mg, 3.70 mmol), and the temperature was raised to 60° C. and stirred for 4 hours. After the reaction was completed, distillation was carried out under reduced pressure to remove part of the methanol. The remaining concentrate was dissolved in aqueous sodium bicarbonate (150 mL), and then extracted with dichloromethane (100 mL×3). The organic phase was collected, washed with saturated brine, and then dried over anhydrous sodium sulfate. The solvent was evaporated in vacuo, and the residue was purified by column chromatography with petroleum ether / ethyl acetate = 3 / 1 as eluent to give the crude product intermediate 1-13 (869 mg, yellow solid).

[0849] ESI-MS: m / z = 555.2, [M+H] + .

[0850] Step 14:1 6 -Br-2 1 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 5 -Chloro-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 1-14)

[0851]

[0852] To a solution of the crude intermediate 1-13 (551 mg, 0.993 mmol) in acetonitrile (15 mL) were added p-toluenesulfonic acid monohydrate (378 mg, 1.98 mmol) and tert-butyl nitrite (153 μl, 1.29 mmol), and the mixture was stirred at room temperature for 0.5 h. Then, copper bromide (55 mg, 0.25 mmol) and tetrabutylammonium bromide (1130 mg, 3.505 mmol) were added, and the mixture was warmed to 0° C. and stirred for 3 h. After the reaction was completed, the mixture was quenched with water (100 mL), and then extracted with dichloromethane (50 mL×3). The organic phase was collected, washed with saturated brine, and dried over anhydrous sodium sulfate. The solvent was evaporated in vacuo, and the residue was purified by column chromatography with petroleum ether / ethyl acetate = 10 / 1 to 4 / 1 as the eluent to give the product intermediate 1-14 (572 mg, yellow liquid) in a yield of 73%.

[0853] ESI-MS: m / z = 620.1, [M+H] + .

[0854] Step 15:1 6 -(4-methylpiperazin-1-yl)-2 1 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 5 -Chloro-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 1-15)

[0855]

[0856] To a solution of intermediate 1-14 (30 mg, 0.048 mmol) in toluene (3 mL) were added N-methylpiperazine (14 mg, 0.14 mmol), palladium acetate (2 mg, 0.009 mmol), BINAP (12 mg, 0.019 mmol), cesium carbonate (81 mg, 0.25 mmol), nitrogen was replaced, the temperature was raised to 100 ° C. and stirred for 14 hours. After the reaction was completed, it was quenched with water (30 mL), and then extracted with dichloromethane (20 mL×3). The organic phase was collected, washed with saturated brine, and then dried over anhydrous sodium sulfate. The solvent was evaporated in vacuo, and the residue was purified by preparative thin layer chromatography using petroleum ether / ethyl acetate = 1 / 1 as the developing solvent to obtain the product intermediate 1-15 (18 mg, white solid), 58%.

[0857] ESI-MS: m / z = 638.3, [M+H] + .

[0858] Step 16: Synthesis of Compound 1

[0859] To a solution of intermediate 1-15 (18 mg, 0.028 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL) and stirred at room temperature for 14 hours. After the reaction was completed, the mixture was dried in vacuo and the residue was dissolved in acetonitrile (4 mL). Ammonia (1 mL) was added and the mixture was reacted at room temperature for 30 minutes. After the reaction was completed, the mixture was dried in vacuo and the residue was dissolved in dichloromethane and purified by preparative thin layer chromatography using dichloromethane / methanol = 12 / 1 as the developing solvent. The crude product was dissolved in methanol and methyl tert-butyl ether was added to slurry to obtain compound 1 (4 mg, white solid) in a yield of 28%.

[0860] ESI-MS: m / z = 508.2, [M+H] + .

[0861] 1 H NMR(DMSO-d6,400MHz)δ13.81(s,1H),9.15(d,J=1.1Hz,1H),8.50(d,J=1.2H z,1H),7.74-7.64(m,2H),7.52(dd,J=8.2,1.8Hz,1H),7.46(dd,J=8.3,1.2H z,1H),7.04(d,J=8.3Hz,1H),4.39(t,J=6.9Hz,2H),4.32(s,2H),3.70(t,J= 5.4Hz,2H),3.06(s,4H),2.52-2.48(m,4H),2.25(s,3H),2.12-2.02(m,2H).

[0862] The following compounds were synthesized by a method similar to Example 1 using appropriate amines:

[0863]

[0864]

[0865] Example 8

[0866] Compound 8: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 H-3 5 -Chloro-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane

[0867]

[0868] Step 1: Preparation of 3-chloro-2-fluoro-6-methoxyphenylboronic acid (Intermediate 8-1)

[0869]

[0870] Under nitrogen, 4-chloro-3-fluoro-anisole (5.00 g, 31.1 mmol) was dissolved in dry tetrahydrofuran (50 mL), cooled to -60°C, and triisopropyl borate (7.02 g, 37.4 mmol) was added. A 2M solution of lithium diisopropylamine in tetrahydrofuran (31 mL, 62 mmol) was added dropwise at -60°C, and the reaction was maintained at low temperature for 1 h. The mixture was heated to 0°C, diluted with water, and the pH was adjusted to about 4 with 3N HCl. The mixture was extracted with ethyl acetate (50 mL × 3), washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain intermediate 8-1 (5.1 g, white solid) in a yield of 80.9%.

[0871] Step 2: Preparation of 3-chloro-2-fluoro-6-hydroxyphenylboronic acid (Intermediate 8-2)

[0872]

[0873] Under nitrogen, intermediate 8-1 (3.00 g, 14.7 mmol) was dissolved in DCM (40 mL), cooled to -30°C, added with boron tribromide (1.4 mL, 29 mmol), and maintained at 0°C for 1 h. Water (20 mL) was added, extracted with ethyl acetate (30 mL × 3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and slurried with petroleum ether (15 mL) to obtain intermediate 8-2 (2.30 g, white solid) in a yield of 82%.

[0874] Step 3: Preparation of (R)-5-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenol (Intermediate 8-3)

[0875]

[0876] This compound was prepared according to the procedure described in Example 31 (Step 2) using 5-bromo-3-iodo-1-(2-(trimethylsilyl)ethoxy)methyl-1H-pyrazolo[3,4-c]pyridine instead of Intermediate 31-1 as the starting material. Yield: 56.8%.

[0877] ESI-MS: m / z = 560.2, [M+H] + .

[0878] Step 4: Preparation of (R)-3-(5-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenoxy)propan-1-ol (Intermediate 8-4)

[0879]

[0880] This compound was prepared according to the procedure described in Example 31 (Step 3) using Intermediate 8-3 instead of Intermediate 31-2 as the starting material. Yield: 79.2%.

[0881] ESI-MS: m / z = 618.3, [M+H] + .

[0882] Step 5: Preparation of (R)-4-chloro-3-fluoro-2-(3-(4-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-hydroxypropyloxy)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)phenol (Intermediate 8-5)

[0883]

[0884] Under nitrogen, intermediate 8-4 (160 mg, 0.26 mmol), intermediate 8-2 (99 mg, 0.52 mmol), Xphos-Pd-G2 (20 mg, 0.030 mmol), and potassium phosphate (166 mg, 0.78 mmol) were dissolved in a mixed solvent of 1,4-dioxane / water (10 / 2 mL) and heated to 85°C for 3 h. The mixture was quenched with water (30 mL) and extracted with ethyl acetate (20 mL × 3). The mixture was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (methanol / ethyl acetate = 1 / 50) to obtain intermediate 8-5 (121 mg, yellow oil) in a yield of 68%.

[0885] ESI-MS: m / z = 684.3, [M+H] + .

[0886] Step 6: Preparation of (R)-4-chloro-3-fluoro-2-(3-(4-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-bromopropoxy)phenyl)-1-((2-(trimethylsilyl))ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)phenol (Intermediate 8-6)

[0887]

[0888] Under nitrogen protection, intermediate 8-5 (91 mg, 0.14 mmol) was dissolved in dichloromethane (5 mL), triphenylphosphine (32 mg, 0.18 mmol) was added, and the mixture was cooled to 0°C. NBS (47 mg, 0.18 mmol) was added and the reaction was allowed to proceed at room temperature for 2 h. The mixture was quenched with water (10 mL), extracted with dichloromethane (20 mL × 3), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (methanol / ethyl acetate = 1 / 50) to obtain intermediate 8-6 (41 mg, yellow oil) in a yield of 39.4%.

[0889] ESI-MS: m / z = 746.3, [M+H] + .

[0890] Step 7: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 -((2-(trimethylsilyl))ethoxy)methyl)-3-((2-(trimethylsilyl ... 5 -Chloro-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane (Intermediate 8-7)

[0891]

[0892] Under nitrogen, intermediate 8-6 (33 mg, 0.050 mmol) was dissolved in DMF (5 mL), and cesium carbonate (50 mg, 0.15 mmol) and sodium iodide (7 mg, 0.05 mmol) were added. The mixture was heated to 90°C for 3 h. Water (10 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (20 mL × 3). The mixture was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by Prep-TLC (developed with ethyl acetate) to give intermediate 8-7 (12 mg, white solid) in a yield of 31.5%.

[0893] ESI-MS: m / z = 666.3, [M+H] + .

[0894] Step 8: Preparation of Compound 8

[0895]

[0896] Under nitrogen protection, intermediate 8-7 (12 mg, 0.02 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added at 0°C, and the reaction was allowed to proceed at room temperature for 18 h. The mixture was concentrated under reduced pressure, acetonitrile (5 mL) and aqueous ammonia (1 mL) were added, and the mixture was stirred at room temperature for 30 min. The mixture was extracted with (dichloromethane / isopropanol = 5 / 1) (10 mL × 3). The combined organic phases were washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by Prep-TLC (dichloromethane / methanol = 10 / 1) to obtain the title compound 8 (7.30 mg, white solid) in a yield of 75%.

[0897] ESI-MS: m / z = 536.3, [M+H] + .

[0898] 1 H NMR(DMSO-d6,400MHz)δ13.67(s,1H),9.10(s,1H),8.78(s,1H),7.82(d,J=1.9Hz,1H), 7.58(t,J=8.7Hz,1H),7.48(dd,J=8.2,1.7Hz,1H),7.146.93(m,2H),4.64-4.44(m,2H), 4.13(s,2H),3.82-3.75(m,1H),3.72-3.65(m,1H),3.61-3.42(m,2H),3.30-3.23(m,1H ), 3.17 (t, J = 10.0Hz, 1H), 2.76 (t, J = 11.7Hz, 2H), 2.70-2.62 (m, 1H), 2.45-2.20 (m, 6H).

[0899] Example 9

[0900] Compound 9: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 H-3 5 -cyclopropyl-3- 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane

[0901]

[0902] Step 1: Preparation of 4-cyclopropyl-3-fluorophenol (Intermediate 9-1)

[0903]

[0904] To a mixed solvent of 1,4-dioxane (20 mL) and toluene (20 mL) were added 4-bromo-3-fluorophenol (2.047 g, 10.7 mmol), cyclopropylboronic acid (2.131 g, 24.8 mmol), and potassium carbonate (6.329 g, 45.8 mmol), and the atmosphere was replaced with nitrogen. Palladium acetate (207 mg, 0.92 mmol) and tricyclohexylphosphine (669 mg, 2.39 mmol) were added to the reaction system, and the mixture was heated to 85°C overnight under nitrogen. The reaction mixture was cooled to room temperature, and ethyl acetate (50 mL) and saturated aqueous ammonium chloride (50 mL) were added. The mixture was filtered through celite, and the mother liquor was allowed to stand for separation. The organic phase was washed with brine, dried, and concentrated. The residue was purified by column chromatography to afford Intermediate 9-1 (1.772 g, light brown liquid) in a quantitative yield.

[0905] ESI-MS: m / z = 151.2, [MH] - .

[0906] Step 2: Preparation of 1-cyclopropyl-2-fluoro-4-(methoxymethoxy)benzene (Intermediate 9-2)

[0907]

[0908] To a solution of intermediate 9-1 (1256 mg, 8.25 mmol) in dichloromethane (25 mL) was added DIPEA (4.313 mL, 24.76 mmol). After stirring at room temperature for 5 minutes, bromomethyl methyl ether (2093 mg, 16.75 mmol) was added dropwise. The mixture was stirred at room temperature for 14 h. After completion of the reaction, water (100 mL) was added to quench the mixture, followed by extraction with dichloromethane (50 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo to obtain the crude product. The crude product was purified by column chromatography using petroleum ether / ethyl acetate = 30 / 1 as eluent to obtain intermediate 9-2 (1257 mg, yellow liquid) in a yield of 78%.

[0909] ESI-MS: m / z = 197.1, [M+H] + .

[0910] Step 3: Preparation of 3-cyclopropyl-2-fluoro-6-(methoxymethoxy)phenylboronic acid (Intermediate 9-3)

[0911]

[0912] Intermediate 9-2 (299 mg, 1.52 mmol) was dissolved in a mixture of dry tetrahydrofuran (5 mL) and dry cyclohexane (5 mL). Under nitrogen, N,N,N',N'-tetramethylethylenediamine (0.40 mL, 2.67 mmol) was added, and the mixture was cooled to -70°C. A 1.3 M solution of sec-butyllithium in n-hexane (2.0 mL, 2.6 mmol) was added dropwise. After addition, the temperature was raised to -55 to -45°C and stirred for one hour. The mixture was cooled again to -65°C and triisopropyl borate (0.64 mL, 2.8 mmol) was added dropwise. After addition, the temperature was slowly raised to 0°C and stirred for 30 minutes. Methanol (5 mL) and 4 M methanolic hydrogen chloride (4 mL) were slowly added to the reaction system, and the mixture was stirred at room temperature overnight. The reaction system was diluted with water and extracted twice with ethyl acetate. The organic phases were combined and concentrated to dryness. The residue was redissolved in methyl tert-butyl ether and extracted twice with 0.15M aqueous sodium hydroxide solution (10 mL*2). The aqueous phases were combined and washed with methyl tert-butyl ether. 3M hydrochloric acid (1 mL) was added dropwise to the aqueous phase until the pH was <3, and the mixture was extracted with ethyl acetate (20 mL*2). The organic phases were combined, dried, and concentrated to dryness to obtain intermediate 9-3 (249 mg, light yellow liquid) in a yield of 68%.

[0913] ESI-MS: m / z = 257.2, [M+OH] - .

[0914] Step 4: Preparation of (3-cyclopropyl-2-fluoro-6-hydroxyphenyl)boronic acid (Intermediate 9-4)

[0915]

[0916] Intermediate 9-3 (249 mg, 1.03 mmol) was dissolved in a mixed solvent of 1,4-dioxane (2 mL) and methanol (1 mL). Hydrochloric acid (0.5 mL) was added dropwise under ice-water cooling. The mixture was warmed to room temperature and stirred overnight. The reaction mixture was diluted with water and extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with water, dried, and concentrated to dryness to obtain Intermediate 9-4 (211 mg, colorless liquid to white solid). The yield was quantitative.

[0917] ESI-MS: m / z = 195.2, [MH] - .

[0918] Compound 9 was prepared according to the preparation method described in Example 8 (steps 5 to 8) using intermediate 9-4 instead of intermediate 8-2 as the starting material.

[0919] ESI-MS: m / z = 542.3, [M+H] + .

[0920] 1H NMR(DMSO-d6,700MHz)δ13.62(s,1H),9.08(s,1H),8.70(s,1H),7.83(s,1H),7.46(d,J=8.0Hz,1H),7.02-6. 90(m,2H),6.85(d,J=8.7Hz,1H),4.52(t,J=8.3Hz,2H),4.04(s,2H),3.77(d,J=10.4Hz,1H),3.69(d,J=10.2H z,1H),3.53(t,J=11.1Hz,1H),3.48(d,J=9.9Hz,1H),3.24(d,J=9.4Hz,1H),3.16(t,J=10.0Hz,1H),2.80-2.7 2(m,2H),2.66(d,J=11.0Hz,1H),2.42-2.22(m,6H),2.08-2.01(m,1H),0.96-0.91(m,2H),0.71-0.66(m,2H).

[0921] Example 10

[0922] Compound 10:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 4 -ethyl-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane

[0923]

[0924] Step 1: Preparation of 2-(3-((5-bromo-3-fluoro-2-iodobenzyl)oxy)propoxy)tetrahydro-2H-pyran (Intermediate 10-1)

[0925]

[0926] This compound was prepared according to the preparation method described in Example 1 (steps 5 to 8) using 5-bromo-3-fluoro-2-iodobenzoic acid instead of Intermediate 1-4 as the starting material.

[0927] ESI-MS: m / z = 495.0, [M+H] + .

[0928] Step 2: Preparation of 5-bromo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine (Intermediate 10-2)

[0929]

[0930] Under nitrogen, 3,4-dihydro-2H-pyran (636 mg, 7.58 mmol) and p-toluenesulfonic acid monohydrate (240 mg, 1.26 mmol) were added to a solution of 5-bromo-1H-pyrazolo[3,4-c]pyridine (500 mg, 2.53 mmol) in 1,4-dioxane (8 mL). The mixture was then heated to 80°C and stirred for 5 hours. After cooling to room temperature, sodium bicarbonate solution (50 mL) was added and the mixture was extracted with ethyl acetate (50 mL x 2). The organic phase was washed once with water (50 mL) and once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 5 / 1 to obtain intermediate 10-2 (670 mg, white solid) in a 94% yield.

[0931] ESI-MS: m / z = 282.1, [M+H] + .

[0932] Step 3: Preparation of 1-(tetrahydro-2H-pyran-2-yl)-5-(tri-n-butyltinyl)-1H-pyrazolo[3,4-c]pyridine (Intermediate 10-3)

[0933]

[0934] Under nitrogen protection, hexabutyl ditin (1.44 mL 2.85 mmol) was added to the 1,4-dioxane (10 mL) system of intermediate 10-2 (670 mg, 2.38 mmol), anhydrous lithium chloride (604 mg 14.2 mmol), tricyclohexylphosphine (66 mg, 0.24 mmol), and Pd2(dba)3 (109 mg, 0.12 mmol), and then the temperature was raised to 100 ° C and stirred for 14 hours. Filtered through celite, the filter cake was washed with ethyl acetate, and the filtrate was collected and evaporated to dryness. The residue was purified by column chromatography with a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to obtain intermediate 10-3 (875 mg, colorless oil) with a yield of 75%.

[0935] ESI-MS: m / z = 494.2, [M+H] + .

[0936] Step 4: Preparation of 5-(4-bromo-2-fluoro-6-((3-((tetrahydro-2H-pyran-2-yl)oxy)propyloxy)methyl)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine (Intermediate 10-4)

[0937]

[0938] Under nitrogen, a solution of intermediate 10-3 (778 mg, 1.58 mmol), intermediate 10-1 (972 mg, 2.06 mmol), and Pd(PPh3)2Cl2 (111 mg, 0.16 mmol) in xylene (15 mL) was heated to 130°C and stirred for 14 hours. The mixture was cooled to room temperature, added with water (60 mL), and extracted with ethyl acetate (60 mL). The organic phase was washed once with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to obtain intermediate 10-4 (503 mg, colorless oil) in a yield of 53%.

[0939] ESI-MS: m / z = 548.1, [M+H] + .

[0940] Step 5: Preparation of 3-((5-bromo-3-fluoro-2-(1H-pyrazolo[3,4-c]pyridin-5-yl)benzyl)oxy)propan-1-ol (Intermediate 10-5)

[0941]

[0942] To a solution of intermediate 10-4 (503 mg, 0.92 mmol) in dichloromethane (5 mL) was added a 4 M solution of hydrogen chloride in 1,4-dioxane (5 mL) dropwise at room temperature, stirred overnight, and rotary evaporated to dryness to afford intermediate 10-5 (348 mg, light yellow oil) in 100% yield.

[0943] ESI-MS: m / z = 380.1, [M+H] + .

[0944] Step 6: Preparation of 3-((5-bromo-3-fluoro-2-(3-iodo-1H-pyrazolo[3,4-c]pyridin-5-yl)benzyl)oxy)propan-1-ol (Intermediate 10-6)

[0945]

[0946] To a solution of intermediate 10-5 (348 mg, 0.92 mmol) in DMAc (30 mL) at 0°C were added potassium hydroxide (154 mg, 2.74 mmol) and iodine (256 mg, 1.00 mmol), followed by stirring at room temperature overnight. The reaction was quenched with aqueous ammonium chloride (50 mL) and extracted with ethyl acetate (50 mL x 2). The organic phase was washed once with aqueous sodium thiosulfate (50 mL), once with water (50 mL), and once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 10-6 (485 mg, yellow solid) was obtained in a 100% yield.

[0947] ESI-MS: m / z = 505.9, [M+H] + .

[0948] Step 7: Preparation of 5-(4-bromo-2-fluoro-6-((3-((tetrahydro-2H-pyran-2-yl)oxy)propyloxy)methyl)phenyl)-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridine (Intermediate 10-7)

[0949]

[0950] Under nitrogen protection, to a solution of intermediate 10-6 (450 mg, 0.89 mmol) in 1,4-dioxane (8 mL) were added p-toluenesulfonic acid monohydrate (84 mg, 0.44 mmol) and 3,4-dihydro-2H-pyran (224 mg, 2.66 mmol), and the temperature was raised to 100 ° C and stirred for 2 hours. After cooling to room temperature, water (50 mL) was added to quench the mixture and the mixture was extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography with a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to obtain intermediate 10-7 (503 mg, yellow solid) with a yield of 81%.

[0951] ESI-MS: m / z = 674.0, [M+H] + .

[0952] Step 8: Preparation of tert-butyl 4-(4-(5-(4-bromo-2-fluoro-6-(3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)methyl)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 10-8)

[0953]

[0954] This compound was prepared according to the procedure described in Example 31 (Step 2) using Intermediate 10-7 (465 mg, 0.69 mmol) instead of Intermediate 31-1 and Intermediate A3 instead of Intermediate A2 as starting materials to obtain Intermediate 10-8 (430 mg, yellow solid) in a yield of 76%.

[0955] ESI-MS: m / z = 824.3, [M+H] + .

[0956] Step 9: Preparation of tert-butyl 4-(4-(5-(4-bromo-2-fluoro-6-((3-hydroxypropyloxy)methyl)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 10-9)

[0957]

[0958] This compound was prepared according to the procedure described in Example 1 (Step 11) using Intermediate 10-8 (461 mg, 0.56 mmol) instead of Intermediate 1-10 as the starting material to obtain Intermediate 10-9 (363 mg, yellow solid) with a yield of 87.7%.

[0959] ESI-MS: m / z = 740.3, [M+H] + .

[0960] Step 10:1 6 -(4-methylpiperazin-1-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 4 -Br-3 6 Preparation of 5,9-dioxa-1(1,3)-benzo-2(3,5)-(6-azaindazole)-3(1,2)-benzheterocyclononane (Intermediate 10-10)

[0961]

[0962] This compound was prepared according to the procedure described in Example 1 (Step 12) using Intermediate 10-9 (363 mg, 0.49 mmol) instead of Intermediate 1-11 as the starting material to obtain Intermediate 10-10 (310 mg, white solid) with a yield of 87.5%.

[0963] ESI-MS: m / z = 722.2, [M+H] + .

[0964] Step 11:1 6 -(4-methylpiperazin-1-yl)-2 1-(tetrahydro-2H-pyran-2-yl)-3 4 -Vinyl-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 10-11)

[0965]

[0966] Under nitrogen, a mixture of intermediate 10-10 (245 mg, 0.34 mmol), vinylboronic acid pinacol ester (157 mg, 1.02 mmol), Pd(dppf)Cl2 (37 mg, 0.05 mmol), potassium phosphate (216 mg, 1.02 mmol) in 1,4-dioxane (12 mL) and water (4 mL) was stirred at 100°C for 14 hours. The mixture was then cooled to room temperature, diluted with water (50 mL), and extracted with ethyl acetate (50 mL x 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to obtain intermediate 10-11 (218 mg, white solid) in a 96% yield.

[0967] ESI-MS: m / z = 670.3, [M+H] + .

[0968] Step 12:1 6 -(4-methylpiperazin-1-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 4 -ethyl-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 10-12)

[0969]

[0970] To a solution of intermediate 10-11 (30 mg, 0.04 mmol) in tetrahydrofuran (2 mL) and methanol (2 mL) was added platinum dioxide (5 mg, 0.02 mmol) at room temperature and stirred under a hydrogen atmosphere for 3 hours. The mixture was filtered through a pad of celite, and the filtrate was collected and evaporated to dryness. Intermediate 10-12 (31 mg, light yellow solid) was obtained in a 100% yield.

[0971] ESI-MS: m / z = 672.4, [M+H] + .

[0972] Step 13:1 6-(piperazin-1-yl)-2 1 H-3 4 -ethyl-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 10-13)

[0973]

[0974] To a solution of intermediate 10-12 (31 mg, 0.04 mmol) in dichloromethane (1 mL) was added a 4M solution of hydrogen chloride in 1,4-dioxane (2 mL) dropwise at room temperature, stirred overnight, and rotary evaporated to dryness to afford crude intermediate 10-13 (21 mg, light yellow solid) in quantitative yield.

[0975] ESI-MS: m / z = 488.3, ​​[M+H] + .

[0976] Step 14: Preparation of compound 10

[0977] At room temperature, 1H-benzotriazole-1-methanol (10 mg, 0.06 mmol), sodium acetate (11 mg, 0.13 mmol), and sodium triacetoxyborohydride (32 mg, 0.15 mmol) were added to a solution of intermediate 10-13 (21 mg, 0.040 mmol) in dichloromethane / methanol = 1 / 1 (3 mL) and stirred at room temperature for 2 hours. The reaction was quenched with sodium bicarbonate solution (15 mL) and extracted with dichloromethane / isopropanol = 5 / 1 (15 mL × 3). The organic phases were combined and washed once with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography eluting with dichloromethane / methanol = 7:1 to give the title product (13.5 mg, white solid) in a 64% yield.

[0978] ESI-MS: m / z = 502.2, [M+H] + .

[0979] 1H NMR(DMSO-d6,400MHz)δ13.75(s,1H),9.13(s,1H),8.45(s,1H),7.71(s,1H) ,7.52(d,J=8.2Hz,1H),7.25(s,1H),7.18(d,J=11.1Hz,1H),7.05(d,J=8.3Hz ,1H),4.40(t,J=6.6Hz,2H),4.31(s,2H),3.75-3.67(m,2H),3.11(br,4H),2. 74-2.60(m,5H),2.42-2.30(s,3H),2.13-2.02(m,2H),1.25(t,J=7.6Hz,3H).

[0980] Example 11

[0981] Compound 11:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 4 -(1-Hydroxyethyl)-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane

[0982]

[0983] Step 1:1 6 -(4-methylpiperazin-1-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 4 -aldehyde-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 11-1)

[0984]

[0985] To a solution of Intermediate 10-11 (210 mg, 0.31 mmol), potassium osmate (3 mg, 0.006 mmol), and 2,6-lutidine (67 mg, 0.63 mmol) in 1,4-dioxane (6 mL) and water (2 mL) was added sodium periodate (268 mg, 1.25 mmol) at room temperature and stirred for 15 hours. The reaction was quenched by the addition of saturated sodium thiosulfate solution (50 mL) and extracted with ethyl acetate (50 mL x 2). The organic phase was washed sequentially with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient of petroleum ether / ethyl acetate (10 / 1 to 2 / 1) to afford Intermediate 11-1 (160 mg, white solid) in a 76% yield.

[0986] ESI-MS: m / z = 672.3, [M+H] + .

[0987] Step 2:1 6 -(4-methylpiperazin-1-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 4 -(1-Hydroxyethyl)-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 11-2)

[0988]

[0989] To a solution of intermediate 11-1 (30 mg, 0.040 mmol) in tetrahydrofuran (2 mL) under nitrogen was added dropwise a 1.4 M solution of methylmagnesium bromide in toluene (0.10 mL, 0.14 mmol) at 0°C, followed by stirring at 0°C for 1 hour. The mixture was quenched with aqueous ammonium chloride and then extracted with ethyl acetate (20 mL x 2). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 11-2 (32 mg, light yellow solid) was obtained in a 100% yield.

[0990] ESI-MS: m / z = 688.4, [M+H] + .

[0991] Compound 11 was prepared according to the preparation method described in Example 10 (steps 13 to 14) using intermediate 11-2 (32 mg, 0.04 mmol) instead of intermediate 10-12 as the starting material to give the title product (15 mg, white solid) with a two-step yield of 71%.

[0992] ESI-MS: m / z = 518.2, [M+H] + .

[0993] 1 H NMR(DMSO-d6,400MHz)δ13.75(s,1H),9.13(s,1H),8.45(s,1H),7.71(s,1H),7.52(d ,J=8.2Hz,1H),7.37(s,1H),7.26(d,J=11.1Hz,1H),7.04(d,J=8.3Hz,1H),5.37(d,J =3.7Hz,1H),4.86-4.76(m,1H),4.40(t,J=6.6Hz,2H),4.33(s,2H),3.76-3.66(m,2H ), 3.10 (s, 4H), 2.63 (br, 4H), 2.34 (s, 3H), 2.13-2.03 (m, 2H), 1.39 (d, J = 6.4Hz, 3H).

[0994] Example 12

[0995] Compound 12:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 4 -(methylaminomethyl)-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane

[0996]

[0997] Step 1-2:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 4 -aldehyde-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 12-2)

[0998]

[0999] Intermediate 12-2 was prepared according to the preparation method described in Example 10 (steps 13 to 14) using intermediate 11-1 (30 mg, 0.04 mmol) instead of intermediate 10-12 as the starting material to give the title product (18 mg, white solid) with a two-step yield of 87.5%.

[1000] ESI-MS: m / z = 502.2, [M+H] + .

[1001] Step 3: Preparation of compound 12

[1002] At room temperature, a 30% methylamine solution in ethanol (50 μL) was added to a solution of intermediate 12-2 (18 mg, 0.036 mmol) in dichloromethane / methanol = 1 / 1 (3 mL). The mixture was heated to 50°C and stirred for 30 minutes. Acetic acid (50 μL) and sodium triacetoxyborohydride (72 mg, 0.32 mmol) were then added and stirred at 50°C for 5 hours. The reaction was quenched with sodium bicarbonate solution (15 mL) and extracted with dichloromethane / isopropanol = 5 / 1 (15 mL × 2). The organic phases were combined and washed once with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 6 / 1 as the eluent to give the title product (8.1 mg, white solid) in a 43% yield.

[1003] ESI-MS: m / z = 517.2, [M+H] + .

[1004] 1 H NMR(DMSO-d6,400MHz)δ13.77(br,1H),9.14(s,1H),8.46(s,1H),7.70(s,1H ),7.52(d,J=8.2Hz,1H),7.44(s,1H),7.39(d,J=11.1Hz,1H),7.04(d,J=8.3 Hz,1H),4.40(t,J=6.6Hz,2H),4.33(s,2H),3.97(s,2H),3.76-3.66(m,2H), 3.06(s,4H),2.55-2.50(m,4H),2.47(s,3H),2.24(s,3H),2.13-2.03(m,2H).

[1005] Example 13

[1006] Compound 13:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 4 -ethynyl-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane

[1007]

[1008] Step 1:1 6 -(4-methylpiperazin-1-yl)-2 1-(tetrahydro-2H-pyran-2-yl)-3 4 -ethynyl-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 13-1)

[1009]

[1010] Under nitrogen, anhydrous potassium carbonate (18 mg, 0.15 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (17 μL, 0.10 mmol) were added to a solution of intermediate 11-1 (35 mg, 0.05 mmol) in methanol (2 mL). The mixture was stirred at room temperature for 8 hours. The mixture was quenched with water and then extracted with ethyl acetate (20 mL x 2). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. This afforded crude intermediate 13-1 (40 mg, pale yellow solid) in a 100% yield.

[1011] ESI-MS: m / z = 668.3, [M+H] + .

[1012] Compound 13 was prepared according to the preparation method described in Example 10 (steps 13 to 14) using intermediate 13-1 (40 mg, 0.050 mmol) instead of intermediate 10-12 as the starting material to give the title product (14 mg, white solid) with a two-step yield of 53%.

[1013] ESI-MS: m / z = 498.2, [M+H] + .

[1014] 1 H NMR(DMSO-d6,400 MHz)δ13.79(br,1H),9.14(s,1H),8.50(s,1H),7.70(s,1H),7.58(s,1H),7.52(d,J=8.2 Hz,1H),7.47(d,J=11.1 Hz,1H),7.04(d,J=8.3 Hz,1H),4.44-4.36(m,3H),3.74-3.67(m,2H),3.09(s,4H),2.63(br,4H),2.47(s,3H),2.34(s,3H),2.13-2.03(m,2H).

[1015] Example 14

[1016] Compound 14:1 6 -(4-methylpiperazin-1-yl)-21 H-3 4 -Cyano-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane

[1017]

[1018] Step 1:1 6 -(4-methylpiperazin-1-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 4 -Formaldehyde oxime-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 14-1)

[1019]

[1020] To a solution of intermediate 11-1 (35 mg, 0.050 mmol) in ethanol (2 mL) was added triethylamine (30 μL, 0.21 mmol) dropwise at room temperature, followed by hydroxylamine hydrochloride (15 mg, 0.21 mmol). The mixture was stirred at room temperature for 14 hours. After the reaction was complete, water (30 mL) was added and then extracted with ethyl acetate (30 mL x 2). The organic phase was washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 14-1 (36 mg, light yellow solid) was obtained in a 100% yield.

[1021] ESI-MS: m / z = 687.3, [M+H] + .

[1022] Step 2:1 6 -(4-methylpiperazin-1-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 4 -Cyano-3 6 Preparation of 5,9-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclononane (Intermediate 14-2)

[1023]

[1024] To a solution of Intermediate 14-1 (36 mg, 0.05 mmol) in DMSO (2 mL) at room temperature was added potassium carbonate (22 mg, 0.16 mmol), followed by acetic anhydride (15 μL, 0.16 mmol). The mixture was heated to 60°C and stirred for 14 hours. The temperature was cooled to room temperature, water (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL x 2). The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. This afforded crude Intermediate 14-2 (40 mg, light yellow solid) in a quantitative yield.

[1025] ESI-MS: m / z = 669.3, [M+H] + .

[1026] Compound 14 was prepared according to the preparation method described in Example 10 (steps 13 to 14) using intermediate 14-2 (40 mg, 0.050 mmol) instead of intermediate 10-12 as the starting material to give the title product (7 mg, white solid) with a two-step yield of 28%.

[1027] ESI-MS: m / z = 499.2, [M+H] + .

[1028] 1 H NMR(DMSO-d6,400MHz)δ13.88(br,1H),9.17(s,1H),8.54(s,1H),8.02-7.96(m,2H),7.69(s,1H),7.52(d,J=8.2Hz,1H), 7.04(d,J=8.3Hz,1H),4.45-4.35(m,4H),3.76-3.67(m,2H),3.10(s,4H),2.64(br,4H),2.34(s,3H),2.13-2.03(m,2H).

[1029] Example 15

[1030] Compound 15: (R)-6 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-7- ... 1 H-8 6 Preparation of 5,9-dioxa-6(1,3)-benza-7(3,5)-(6-azaindazole)-8(1,2)-benzaspiro[2.7]decane

[1031]

[1032] Step 1: Preparation of (2-bromo-5-iodophenol) acetate (Intermediate 15-1)

[1033]

[1034] Under nitrogen, 2-bromo-5-iodophenol (4.32 g, 14.4 mmol) was dissolved in 1,4-dioxane (50 mL), and pyridine (5.72 g, 72.26 mmol) was added. Acetic anhydride (1.72 g, 17.3 mmol) was added at 0°C, and the reaction was stirred at room temperature for 18 h. The mixture was quenched with water (20 mL), extracted with ethyl acetate (50 mL × 3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (petroleum ether / ethyl acetate = 20 / 1) to obtain intermediate 15-1 (5.01 g, white solid) in a yield of 100%.

[1035] Step 2: Preparation of 3-acetoxy-4-bromophenylboronic acid pinacol ester (Intermediate 15-2)

[1036]

[1037] Under nitrogen, Intermediate 15-1 (3.000 g, 8.80 mmol) was dissolved in 1,4-dioxane (45 mL). Bis(boron)pinacolatoate (2.342 g, 9.24 mmol), Pd(dppf)Cl2 (644 mg, 0.09 mmol), and potassium pivalate (3.701 g, 26.39 mmol) were added at room temperature. The mixture was heated to 90°C for 18 h. Water (50 mL) was added, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified on a silica gel column (petroleum ether / ethyl acetate = 10 / 1) to obtain Intermediate 15-2 (2.702 g, yellow oil) in a yield of 70.0%.

[1038] ESI-MS: m / z=358.2, [M+NH4] + .

[1039] Step 3: Preparation of 2-(3-(4-bromo-3-hydroxyphenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-5-yl)-3-fluorophenol (Intermediate 15-3)

[1040]

[1041] Under nitrogen, Intermediate A1 (432 mg, 0.98 mmol) was dissolved in a 1,4-dioxane / water mixture (8 / 1 mL). Intermediate 15-2 (367 mg, 1.08 mmol), Pd(dppf)Cl2 (72 mg, 0.098 mmol), and potassium phosphate (624 mg, 2.94 mmol) were added, and the mixture was heated to 90°C for 3 h. Aqueous potassium hydroxide (10 mL) was added, and the mixture was stirred for 2 h. The pH was adjusted to 7 with saturated citric acid solution, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified on a silica gel column (petroleum ether / ethyl acetate = 3 / 1) to obtain Intermediate 15-3 (283 mg, yellow oil) in a yield of 59.7%.

[1042] ESI-MS: m / z = 484.2, [M+H] + .

[1043] Step 4:6 6 -Br-7 1 -(tetrahydro-2H-pyran-2-yl)-8 6 Preparation of 5,9-fluoro-6(1,3)-benza-7(3,5)-(6-azaindazole)-8(1,2)-benzaspiro[2.7]decane (Intermediate 15-4)

[1044]

[1045] Under nitrogen, intermediate 15-3 (283 mg, 0.58 mmol) was dissolved in acetonitrile (20 mL), and 1,1-bis(bromomethyl)cyclopropane (132 mg, 0.58 mmol), cesium carbonate (566 mg, 1.74 mmol), and sodium iodide (44 mg, 0.29 mmol) were added. The mixture was heated to 70°C for 3 h. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified on a silica gel column (petroleum ether / ethyl acetate = 2 / 1) to obtain intermediate 15-4 (212 mg, yellow oil) in a yield of 66.1%.

[1046] ESI-MS: m / z = 550.2, [M+H] + .

[1047] Step 5: (R)-6 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-7- ... 1 -(tetrahydro-2H-pyran-2-yl)-8 6Preparation of 5,9-fluoro-6(1,3)-benza-7(3,5)-(6-azaindazole)-8(1,2)-benzaspiro[2.7]decane (Intermediate 15-5)

[1048]

[1049] Under nitrogen, intermediate 15-4 (50 mg, 0.09 mmol) was dissolved in toluene (5 mL), and (R)-octahydropyrazino[2,1-c][1,4]oxazine (51 mg, 0.36 mmol), palladium acetate (2 mg, 0.01 mmol), BINAP (12 mg, 0.02 mmol), and cesium carbonate (88 mg, 0.27 mmol) were added. The mixture was heated to 110°C for 18 h. Water (10 mL) was added, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified on a silica gel column (ethyl acetate = 100%) to obtain intermediate 15-5 (21 mg, yellow oil) in a yield of 42.8%.

[1050] ESI-MS: m / z = 612.5, [M+H] + .

[1051] Step 6: Preparation of compound 15

[1052]

[1053] Under nitrogen, intermediate 15-5 (21 mg, 0.034 mmol) was dissolved in dichloromethane (1 mL), and a 4 M solution of hydrogen chloride in dioxane (2 mL) was added at 0°C. The mixture was allowed to react at room temperature for 18 h. The mixture was concentrated under reduced pressure, adjusted to pH 8 with saturated sodium bicarbonate solution, and stirred at room temperature for 30 min. The mixture was extracted with a mixed solvent of (dichloromethane / isopropanol = 5 / 1) (10 mL x 3). The organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Prep-TLC (dichloromethane / methanol = 10 / 1) yielded the title compound (4 mg, white solid) in a yield of 22.2%.

[1054] ESI-MS: m / z = 528.5, [M+H] + .

[1055] 1H NMR(DMSO-d6,400MHz)δ13.60(s,1H),9.08(s,1H),8.78(s,1H),7.91(s,1H),7.47(dd,J= 8.2,1.4Hz,1H),7.42-7.34(m,1H),7.02-6.92(m,3H),4.71(s,2H),4.02(br,2H),3.77(d, J=10.4Hz,1H),3.68(d,J=10.2Hz,1H),3.54(t,J=11.1Hz,1H),3.41(d,J=9.9Hz,1H),3.2 4-3.12(m,2H),2.80-2.72(m,2H),2.66(d,J=11.0Hz,1H),2.42-2.22(m,4H),0.59(s,4H).

[1056] The following compounds were synthesized by a method similar to Example 15 using appropriate intermediates:

[1057]

[1058] Example 17

[1059] Compound 17: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 H-3 6 Preparation of 6,6-difluoro-1(1,3)-benzo-2(3,5)-(6-azaindazole)-3(1,2)-benzeneheterocyclooctane

[1060]

[1061] Step 1: Preparation of 2,2-difluoro-1,3-propanediol mono-p-toluenesulfonate (Intermediate 17-1)

[1062]

[1063] Under nitrogen protection and at 0°C, DIPEA (3.6 mL, 21 mmol) and p-toluenesulfonyl chloride (2.041 g, 10.71 mmol) were added to a solution of 2,2-difluoro-1,3-propanediol (1.200 g, 10.71 mmol) in dichloromethane (20 mL), and then stirred at room temperature for 16 hours. Water (100 mL) was added to the system and extracted with dichloromethane (100 mL). The organic phase was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography eluting with petroleum ether / ethyl acetate = 5 / 1 to obtain intermediate 17-1 (520 mg, colorless oil) with a yield of 18%.

[1064] ESI-MS: m / z=284.1, [M+NH4] + .

[1065] Step 2: Preparation of (R)-3-(5-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenoxy)-2,2-difluoropropan-1-ol (Intermediate 17-2)

[1066]

[1067] Under nitrogen, cesium carbonate (486 mg, 1.49 mmol) and sodium iodide (37 mg, 0.25 mmol) were added to a solution of intermediate 8-3 (279 mg, 0.50 mmol) and intermediate 17-1 (159 mg, 0.60 mmol) in acetonitrile (10 mL). The mixture was heated to 70°C and stirred for 14 hours. After cooling to room temperature, water (50 mL) was added to the reaction system and extracted with ethyl acetate (50 mL × 2). The organic phases were combined and washed once with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to 1 / 1 as eluent to obtain intermediate 17-2 (120 mg, yellow oil) in a 36% yield.

[1068] ESI-MS: m / z = 654.2, [M+H] + .

[1069] Step 3: Preparation of (R)-2-(3-(3-(2,2-difluoro-3-hydroxypropoxy)-4-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)-3-fluorophenol (Intermediate 17-3)

[1070]

[1071] Under nitrogen, a solution of Intermediate 17-2 (120 mg, 0.18 mmol), 2-fluoro-6-hydroxyphenylboronic acid (57 mg, 0.37 mmol), potassium phosphate (117 mg, 0.55 mmol), and Xphos-Pd-G2 (15 mg, 0.02 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was heated to 80°C and stirred for 2 hours. The mixture was cooled to room temperature, diluted with water (30 mL), and extracted with ethyl acetate (30 mL x 2). The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to pure ethyl acetate to obtain Intermediate 17-3 (125 mg, yellow solid) in 100% yield.

[1072] ESI-MS: m / z = 686.3, [M+H] + .

[1073] Step 4: Preparation of (R)-2-(3-(3-(2,2-difluoro-3-iodopropoxy)-4-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-5-yl)-3-fluorophenol (Intermediate 17-4)

[1074]

[1075] Under nitrogen, to a solution of Intermediate 17-3 (90 mg, 0.13 mmol) in tetrahydrofuran (5 mL) were added triphenylphosphine (177 mg, 0.66 mmol), imidazole (45 mg, 0.66 mmol), and iodine (167 mg, 0.66 mmol). The mixture was heated to 80°C and stirred for 4 hours. The mixture was cooled to 0°C and quenched with aqueous sodium thiosulfate. The mixture was extracted with ethyl acetate (30 mL x 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to pure ethyl acetate to obtain crude Intermediate 17-4 (226 mg, white solid) in a quantitative yield.

[1076] ESI-MS: m / z = 796.2, [M+H] + .

[1077] Step 5: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-21 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 6 Preparation of 4,8-dioxa-6,6-difluoro-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane (Intermediate 17-5)

[1078]

[1079] To a solution of Intermediate 17-4 (226 mg, 0.28 mmol) in DMF (10 mL) was added cesium carbonate (259 mg, 0.84 mmol) at room temperature, and the mixture was heated to 100°C and stirred for 5 hours. The mixture was cooled to room temperature, quenched with water (50 mL), and extracted with ethyl acetate (50 mL x 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. This afforded the crude product of Intermediate 17-5 (220 mg, white solid) in a quantitative yield.

[1080] ESI-MS: m / z = 668.3, [M+H] + .

[1081] Step 6: Preparation of compound 17

[1082] To a solution of intermediate 17-5 (220 mg, 0.33 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (1.0 mL) at room temperature, followed by stirring at room temperature for 14 hours. The system was rotary evaporated to dryness. The crude product was redissolved in acetonitrile (5 mL), and aqueous ammonia (0.5 mL) was added thereto and stirred for 30 minutes. Aqueous sodium carbonate solution (20 mL) was added, extracted with dichloromethane (20 mL × 3), the organic phases were combined and washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 10 / 1 as eluent to give the title product (17 mg, white solid) with a three-step yield of 24%.

[1083] ESI-MS: m / z = 538.2, [M+H] + .

[1084] 1H NMR(DMSO-d6,400MHz)δ13.67(s,1H),9.11(s,1H),8.69(s,1H),7.76(s,1H),7.49(dd, J=8.2,1.4Hz,1H),7.47-7.41(m,1H),7.12-7.01(m,3H),5.00(br,2H),4.49(br,2H),3 .78(d,J=10.4Hz,1H),3.71(d,J=10.2Hz,1H),3.60-3.48(m,2H),3.27(d,J=10.2Hz,1H ), 3.18 (t, J = 10.3Hz, 1H), 2.85-2.76 (m, 2H), 2.68 (d, J = 11.0Hz, 1H), 2.46-2.24 (m, 4H).

[1085] Example 18

[1086] Compound 18:1 6 -(1-oxa-8-azaspiro[4.5]dec-8-yl)-2 1 H-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane

[1087]

[1088] Step 1: Preparation of 3-(2-bromo-5-iodophenoxy)propan-1-ol (Intermediate 18-1)

[1089]

[1090] To a solution of 2-bromo-5-iodophenol (6.000 g, 20.07 mmol) in DMF (60 mL) at room temperature was added anhydrous potassium carbonate (5.540 g, 40.15 mmol), followed by the dropwise addition of 3-bromopropanol (3.627 g, 26.10 mmol). The mixture was heated to 70°C and stirred for 1 hour. The temperature was cooled to room temperature, and water (200 mL) was added to the reaction system. The mixture was extracted with ethyl acetate (200 mL x 2). The organic phases were combined and washed once with water (200 mL) and once with saturated brine (200 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 18-1 (7.164 g, white solid) was obtained in a 100% yield.

[1091] ESI-MS: m / z = 379.0, [M+Na] + .

[1092] Step 2: Preparation of 3-(2-bromo-5-iodophenoxy)propan-1-ol acetate (Intermediate 18-2)

[1093]

[1094] To a solution of intermediate 18-1 (7.164 g, 20.07 mmol) in dichloromethane (100 mL) was added pyridine (4.9 mL, 60 mmol) and acetic anhydride (2.458 g, 24.08 mmol) at room temperature and stirred for 1 hour. The mixture was diluted with dichloromethane (100 mL), and the organic phase was washed with 5% citric acid solution (100 mL), washed once with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to obtain intermediate 18-2 (7.538 g, colorless oil) in a 94% yield.

[1095] ESI-MS: m / z=416.0, [M+NH4] + .

[1096] Step 3: Preparation of (3-(3-acetoxypropoxy)-4-bromophenyl)boronic acid pinacol ester (Intermediate 18-3)

[1097]

[1098] Under nitrogen protection, intermediate 18-2 (7.538 g, 18.89 mmol), bis-boronic acid pinacol

[1099] A solution of ester (5.038 g, 19.84 mmol), potassium pivalate (7.947 g, 56.67 mmol), and Pd(dppf)Cl2 (1.382 g, 1.89 mmol) in 1,4-dioxane (100 mL) was heated to 80°C and stirred for 14 hours. The mixture was cooled to room temperature, and most of the 1,4-dioxane was removed by rotary evaporation. The reaction was quenched with water (200 mL) and extracted with ethyl acetate (150 mL x 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to obtain intermediate 18-3 (6.96 g, brown solid) in a 92.3% yield.

[1100] ESI-MS: m / z=416.0, [M+NH4] + .

[1101] Step 4: Preparation of (3-(2-bromo-5-(5-(2-fluoro-6-hydroxyphenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-3-yl)phenoxy)propan-1-ol) acetate (Intermediate 18-4)

[1102]

[1103] Under nitrogen, a mixture of Intermediate A1 (6.76 g, 15.39 mmol), Intermediate 18-3 (6.449 g, 16.16 mmol), Pd(dppf)Cl2 (1.126 g, 1.54 mmol), potassium phosphate (9.800 g, 46.17 mmol) in 1,4-dioxane (100 mL) and water (20 mL) was stirred at 80°C for 4 hours. The mixture was then cooled to room temperature, and water (250 mL) was added. The mixture was extracted with ethyl acetate (150 mL x 2). The organic phase was washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 3 / 1 to obtain Intermediate 18-4 (6.4 g, yellow oil) in a 71% yield.

[1104] ESI-MS: m / z = 584.3, [M+H] + .

[1105] Step 5: Preparation of 2-(3-(4-bromo-3-(3-hydroxypropyloxy)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-5-yl)-3-fluorophenol (Intermediate 18-5)

[1106]

[1107] At 0°C, the intermediate 18-4 (6.400 g, 10.95 mmol) in tetrahydrofuran (60 mL) was added.

[1108] 2M aqueous lithium hydroxide solution (60 mL) was added dropwise to the system and stirred at room temperature for 14 hours. The pH was adjusted to 7-8 with 5% aqueous citric acid. The mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 2). The organic phase was washed once with water (150 mL) and once with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 18-5 (5.620 g, yellow solid) was obtained in a 94.6% yield.

[1109] ESI-MS: m / z = 542.2, [M+H] + .

[1110] Step 6: Preparation of 2-(3-(4-bromo-3-(3-bromopropyloxy)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazolo[3,4-c]pyridin-5-yl)-3-fluorophenol (Intermediate 18-6)

[1111]

[1112] This compound was prepared according to the procedure described in Example 8 (Step 6) using Intermediate 18-5 instead of Intermediate 8-5 as the starting material to give Intermediate 18-6 (5.30 g, white solid) with a yield of 84.2%.

[1113] ESI-MS: m / z = 606.0, [M+H] + .

[1114] Step 7:1 6 -Br-2 1 -(tetrahydro-2H-pyran-2-yl)-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane (Intermediate 18-7)

[1115]

[1116] To a solution of intermediate 18-6 (5.300 g, 8.75 mmol) in acetonitrile (250 mL) was added cesium carbonate (8.558 g, 26.27 mmol) and sodium iodide (676 mg, 4.38 mmol) at room temperature, and the mixture was heated to 70°C and stirred for 2 hours. The mixture was cooled to room temperature, quenched with water (400 mL), and extracted with ethyl acetate (250 mL × 2). The organic phase was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to give intermediate 18-7 (3.70 g, white solid) in a yield of 80.6%.

[1117] ESI-MS: m / z = 524.3, [M+H] + .

[1118] Step 8:1 6 -(1-oxa-8-azaspiro[4.5]dec-8-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane (Intermediate 18-8)

[1119]

[1120] This compound was prepared according to the procedure described in Example 15 (Step 5) using 1-oxa-8-azaspiro[4.5]decane instead of (R)-octahydropyrazino[2,1-c][1,4]oxazine as the starting material to give Intermediate 18-8 in 54% yield.

[1121] ESI-MS: m / z = 585.3, [M+H] + .

[1122] Step 9: Synthesis of compound 18

[1123] This compound was prepared according to the procedure described in Example 15 (Step 6) using Intermediate 18-8 instead of Intermediate 15-5 as the starting material to give the title compound in 54% yield.

[1124] ESI-MS: m / z = 501.3, [M+H] + .

[1125] 1 H NMR(DMSO-d6,400MHz)δ13.61(s,1H),9.09(s,1H),8.75(s,1H),7.84(s,1H ),7.46(d,J=8.1Hz,1H),7.40(q,J=6.8Hz,1H),7.04(d,J=8.2Hz,1H),7.00 -6.93(m,2H),4.60-4.50(m,2H),4.14-4.07(m,2H),3.75(t,J=6.7Hz,2H), 3.12-3.02(m,4H),2.38-2.28(m,2H),1.93-1.85(m,2H),1.76-1.66(m,6H).

[1126] The following compounds were synthesized by using appropriate amines and intermediates in a manner similar to Example 18:

[1127]

[1128]

[1129]

[1130] Example 27

[1131] Compound 27: (R)-1 5 -Fluorine-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 H-36 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane

[1132]

[1133] Step 1: Preparation of 3-(2-bromo-3-fluorophenoxy)propan-1-ol (Intermediate 27-1)

[1134]

[1135] Under nitrogen, 2-bromo-3-fluorophenol (2.0 g, 10 mmol) was dissolved in DMF (20 mL), and potassium carbonate (4.3 g, 31 mmol) and 3-bromo-1-propanol (1.46 g, 13.6 mmol) were added. The mixture was heated to 80°C for 3 h. Water (10 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (20 mL × 3). The mixture was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (petroleum ether / ethyl acetate = 1 / 1) to obtain intermediate 27-1 (2.5 g, colorless oil) in a yield of 96%.

[1136] Step 2: Preparation of 2-(3-(2-bromo-3-fluorophenoxy)propoxy)tetrahydro-2H-pyran (Intermediate 27-2)

[1137]

[1138] Under nitrogen, Intermediate 27-1 (2.5 g, 10 mmol) was dissolved in dichloromethane (25 mL). 3,4-Dihydro-2H-pyran (1.01 g, 12.0 mmol) and pyridine p-toluenesulfonate (705 mg, 2.01 mmol) were added at 0°C and reacted for 3 h at room temperature. The mixture was quenched with water (50 mL) and extracted with dichloromethane (100 mL × 3). The mixture was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (petroleum ether / ethyl acetate = 10 / 1) to obtain Intermediate 27-2 (3.3 g, colorless oil) in a yield of 98%.

[1139] Step 3: Preparation of (9aR)-8-(2-fluoro-6-(3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)phenyl)octahydropyrazino[2,1-c][1,4]oxazine (Intermediate 27-3)

[1140]

[1141] Under nitrogen protection, intermediate 27-2 (3.3 g, 9.9 mmol) was dissolved in toluene (50 mL), and (R)-octahydropyrazino[2,1-c][1,4]oxazine (4.2 g, 30 mmol), Pd2(dba)3 (907 mg, 0.99 mmol), sodium tert-butoxide (2.38 g, 24.8 mmol), and BINAP (1.23 g, 1.98 mmol) were added. The reaction was heated to 110 ° C for 18 h, and water (100 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (150 mL × 3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column (petroleum ether / ethyl acetate = 1 / 1) to obtain the crude product of intermediate 27-3 (3.52 g, yellow oil).

[1142] ESI-MS: m / z = 395.3, [M+H] + .

[1143] Step 4: Preparation of (R)-3-(3-fluoro-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenoxy)propan-1-ol (Intermediate 27-4)

[1144]

[1145] Under nitrogen, the crude intermediate 27-3 (3.52 g, 8.92 mmol) was dissolved in methanol (40 mL). p-Toluenesulfonic acid monohydrate (849 mg, 4.46 mmol) was added at 0°C and the reaction was allowed to proceed for 4 h at room temperature. The system was poured into a saturated NaHCO solution, extracted with dichloromethane (100 mL × 3), washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (petroleum ether / ethyl acetate = 1 / 2) to obtain intermediate 27-4 (1.01 g, yellow oil) with a two-step yield of 32.9%.

[1146] ESI-MS: m / z = 311.2, [M+H] + .

[1147] Step 5: Preparation of (R)-(3-(3-fluoro-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenoxy)propan-1-ol) acetate (Intermediate 27-5)

[1148]

[1149] Under nitrogen protection, intermediate 27-4 (1.01 g, 3.26 mmol) was dissolved in 1,2-dichloroethane (20

[1150] mL), pyridine (1.29 g, 16.3 mmol) was added at 0°C, followed by acetic anhydride (399 mg, 3.91 mmol), and the mixture was heated to 40°C for 8 h. Water (100 mL) was added, and the mixture was extracted with dichloromethane (100 mL × 3). The mixture was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified on a silica gel column (dichloromethane / methanol = 30 / 1) to obtain intermediate 27-5 (1.02 g, yellow oil) in a yield of 88.9%.

[1151] ESI-MS: m / z = 353.2, [M+H] + .

[1152] Step 6: Preparation of (R)-(3-(5-bromo-3-fluoro-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenoxy)propan-1-ol) acetate (Intermediate 27-6)

[1153]

[1154] Under nitrogen, intermediate 27-5 (650 mg, 1.85 mmol) was dissolved in 48% HBr in acetic acid (5 mL), cooled to 10°C, and bromine (0.3 mL) was added dropwise. The mixture was allowed to warm to room temperature and reacted for 0.5 h. Water (10 mL), sodium sulfite solution (30 mL), and extraction with dichloromethane (40 mL × 3) were added. The mixture was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse medium-pressure preparative chromatography (acetonitrile / water = 1 / 1) to obtain intermediate 27-6 (362 mg, yellow oil) in a yield of 45%.

[1155] ESI-MS: m / z = 431.2, [M+H] + .

[1156] Step 7: Preparation of (R)-3-(3-acetoxypropoxy)-5-fluoro-4-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenylboronic acid pinacol ester (Intermediate 27-7)

[1157]

[1158] Under nitrogen protection, intermediate 27-6 (362 mg, 0.84 mmol) was dissolved in dioxane (10 mL), and bisboron pinacol ester (320 mg, 1.26 mmol), Pd(dppf)Cl2 (62 mg, 0.08 mmol), and potassium acetate (247 mg, 2.52 mmol) were added. The reaction mixture was heated to 90 ° C for 18 h, quenched by adding water (50 mL), extracted with ethyl acetate (50 mL × 3), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column (dichloromethane / methanol = 30 / 1) to obtain intermediate 27-7 (412 mg, yellow oil) in a yield of 100%.

[1159] ESI-MS: m / z = 479.3, [M+H] + .

[1160] Step 8: Preparation of (R)-3-(5-(5-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[3,4-c]pyridin-3-yl)-3-fluoro-2-(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)phenoxy)propyl acetate (Intermediate 27-8)

[1161]

[1162] This compound was prepared according to the procedure described in Example 31 (Step 2) using Intermediate 27-7 instead of Intermediate A2 and 5-bromo-3-iodo-1-(2-(trimethylsilyl)ethoxy)methyl-1H-pyrazolo[3,4-c]pyridine instead of Intermediate 31-1 as the starting material.

[1163] ESI-MS: m / z = 678.3, [M+H] + .

[1164] Compound 27 was prepared according to the preparation method described in Example 8 (steps 5 to 8) using 2-fluoro-6-hydroxyphenylboronic acid instead of intermediate 8-2 and intermediate 27-8 instead of intermediate 8-4 as starting materials.

[1165] ESI-MS: m / z = 520.3, [M+H] + .

[1166] 1H NMR(DMSO-d6,400MHz)δ13.76(s,1H),9.11(s,1H),8.75(s,1H),7.71(s,1H),7.44-7.37(m,1H),7.2 8(d,J=12.0Hz,1H),7.00-6.94(m,2H),4.61-4.53(m,2H),4.14-4.08(m,2H),3.76(d,J=10.4Hz,1H), 3.64(dd,J=10.6,2.1Hz,1H),3.57-3.50(m,1H),3.35-3.26(m,1H),3.13(t,J=10.0Hz,1H),3.06(d, J=11.5Hz,1H),2.95-2.82(m,2H),2.72(d,J=10.8Hz,1H),2.65(d,J=11.0Hz,1H),2.38-2.22(m,5H).

[1167] The following compounds were synthesized by a method similar to Example 27 using appropriate intermediates:

[1168]

[1169]

[1170] Example 30

[1171] Compound 30:1 6 -(4-methylpiperazin-1-yl)-2 1 H-2 6 -Cyano-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-indazol-3(1,2)-benzheterocyclooctane

[1172]

[1173] Step 1: Preparation of tert-butyl 6-bromo-5-chloro-1H-indazole-1-carboxylate (Intermediate 30-1)

[1174]

[1175] To a solution of 6-bromo-5-chloro-1H-indazole (4.65 g, 20.1 mmol) in 1,4-dioxane (20 mL) were added DIPEA (5.1 mL, 30 mmol), di-tert-butyl dicarbonate (4.83 g, 22.1 mmol), and DMAP (20 mg, 0.16 mmol), and the mixture was stirred at room temperature for 2 hours. The reaction was quenched with water and extracted with ethyl acetate. The organic phase was washed once with water and once with brine, dried over anhydrous sodium sulfate, filtered, and rotary evaporated to dryness. The residue was purified by column chromatography using a gradient of petroleum ether / ethyl acetate = 10 / 1 to obtain intermediate 30-1 (6.45 g, white solid) in a 96.8% yield.

[1176] Step 2: Preparation of 5-chloro-1H-indazole-6-carbonitrile (Intermediate 30-2)

[1177]

[1178] Under nitrogen, a solution of intermediate 30-1 (6.41 g, 19.4 mmol), zinc cyanide (2.728 g, 23.23 mmol), Pd2(dba)3 (532 mg, 0.58 mmol), 1,1'-bis(diphenylphosphino)ferrocene (1.037 g, 1.936 mmol), and zinc powder (253 mg, 3.87 mmol) in DMAc (100 mL) was heated to 110°C and stirred for 4 hours. The mixture was cooled to room temperature, diluted with water (400 mL), and extracted with ethyl acetate (200 mL × 2). The organic phase was washed with water (200 mL) and saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 1 / 1 to give intermediate 30-2 (3.005 g, light yellow solid) in a yield of 84.9%.

[1179] ESI-MS: m / z = 178.1, [M+H] + .

[1180] Step 3: Preparation of 2-iodo-5-chloro-1H-indazole-6-carbonitrile (Intermediate 30-3)

[1181]

[1182] To a solution of intermediate 30-2 (3.005 g, 16.92 mmol) in DMAc (30 mL) was added potassium hydroxide (2.848 g, 50.76 mmol) and iodine (4.513 g, 17.76 mmol) at 0°C, followed by stirring at room temperature overnight. The reaction was quenched with aqueous ammonium chloride and extracted with ethyl acetate (150 mL x 2). The organic phase was washed with water (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to afford intermediate 30-3 (3.567 g, white solid) in 70% yield.

[1183] ESI-MS: m / z = 302.0, [M+H] + .

[1184] Step 4: Preparation of 5-chloro-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-6-carbonitrile (Intermediate 30-4)

[1185]

[1186] Under nitrogen, to a solution of intermediate 30-3 (3.405 g, 11.22 mmol) in 1,4-dioxane (50 mL) was added p-toluenesulfonic acid monohydrate (1.067 g, 5.61 mmol) and dihydropyran (2.90 g, 33.7 mmol), and the mixture was heated to 100°C and stirred for 2 hours. The mixture was cooled to room temperature, quenched with water (300 mL), and extracted with ethyl acetate (100 mL × 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 20 / 1 to 5 / 1 to obtain intermediate 30-4 (4.43 g, yellow solid) in a yield of 102%.

[1187] ESI-MS: m / z = 410.0, [M+Na] + .

[1188] Step 5: Preparation of tert-butyl 4-(4-(5-chloro-6-cyano-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 30-5)

[1189]

[1190] Under nitrogen, a mixture of intermediate 30-4 (2.460 g, 6.35 mmol), intermediate A3 (2.830 g, 6.35 mmol), Pd(dppf)Cl2 (464 mg, 0.64 mmol), potassium phosphate (4.040 g, 19.04 mmol), 1,4-dioxane (30 mL) and water (6 mL) was stirred at 80°C for 4 hours. The reaction solution was cooled to 0°C, and a 2M aqueous solution of lithium hydroxide (30 mL) was added dropwise to the system and stirred for 1 hour. The pH was adjusted to 7-8 with a 5% aqueous solution of citric acid. The mixture was diluted with water (200 mL) and extracted with ethyl acetate (150 mL × 2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to 2 / 1 to afford Intermediate 30-5 (2.79 g, yellow solid) in a yield of 81.6%.

[1191] ESI-MS: m / z = 538.2, [M+H] + .

[1192] Step 6: Preparation of 4-(2-bromo-3-fluorophenoxy)butan-1-ol (Intermediate 30-6)

[1193]

[1194] At room temperature, potassium carbonate (2.190 g, 15.70 mmol) and 4-bromobutanol (80% tetrahydrofuran solution, 1.652 g, 8.64 mmol) were added to a solution of 2-bromo-3-fluorophenol (1.500 g, 7.85 mmol) in DMF (15 mL). The mixture was heated to 70° C. and stirred overnight. The system was cooled to room temperature, quenched with water (50 mL), and extracted with ethyl acetate (50 mL×2). The organic phase was washed once with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. A gradient elution with petroleum ether / ethyl acetate = 10 / 1 to 5 / 1 was used to obtain intermediate 30-6 (245 mg, colorless oil) in an 11.8% yield.

[1195] ESI-MS: m / z = 285.0, [M+Na] + .

[1196] Step 7: Preparation of 2-(4-(2-bromo-3-fluorophenoxy)butoxy)tetrahydro-2H-pyran (Intermediate 30-7)

[1197]

[1198] To a solution of intermediate 30-6 (245 mg, 0.93 mmol) in dichloromethane (5 mL) were added 3,4-dihydro-2H-pyran (110 μL, 1.21 mmol) and p-toluenesulfonic acid pyridinium salt (47 mg, 0.19 mmol) and stirred at room temperature for 14 hours. Sodium carbonate solution (40 mL) was then added and the mixture was extracted with dichloromethane (40 mL). The organic phase was washed once with water (40 mL) and once with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 10 / 1 to obtain intermediate 30-7 (286 mg, colorless oil) in an 88.5% yield.

[1199] ESI-MS: m / z = 369.1, [M+Na] + .

[1200] Step 8: Preparation of 2-fluoro-6-(4-((tetrahydro-2H-pyran-2-yl)oxy)butoxy)phenylboronic acid pinacol ester (Intermediate 30-8)

[1201]

[1202] Under nitrogen, a solution of Intermediate 30-7 (286 mg, 0.82 mmol) in dry tetrahydrofuran (4 mL) was cooled to -70°C. A 2.5 M solution of n-butyllithium in n-hexane (0.43 mL, 1.07 mmol) was then slowly added dropwise, maintaining the internal temperature below -60°C. After the addition was complete, the mixture was stirred at -65°C for 1 hour. Isopropyl borate-pinacol ester (245 mg, 1.32 mmol) was then slowly added dropwise, and the mixture was stirred at -65°C for 1 hour. The reaction was quenched with ammonium chloride solution (40 mL) and extracted with ethyl acetate (40 mL x 2). The organic phase was washed once with water (50 mL) and once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. This afforded the crude product of Intermediate 30-8 (366 mg, pale yellow oil) in a 100% yield. The crude product was used directly in the next step without further purification.

[1203] ESI-MS: m / z=412.2, [M+NH4] + .

[1204] Step 9: Preparation of tert-butyl 4-(4-(6-cyano-5-(2-fluoro-6-(4-((tetrahydro-2H-pyran-2-yl)oxy)butoxy)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 30-9)

[1205]

[1206] Under nitrogen, a solution of Intermediate 30-5 (200 mg, 0.37 mmol), Intermediate 30-8 (220 mg, 0.56 mmol), potassium phosphate (237 mg, 1.12 mmol), and Xphos-Pd-G2 (29 mg, 0.04 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was heated to 80°C and stirred for 2 hours. The mixture was cooled to room temperature, diluted with water (30 mL), and extracted with ethyl acetate (40 mL × 2). The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to 2 / 1 to obtain Intermediate 30-9 (262 mg, white solid) in a 90% yield.

[1207] ESI-MS: m / z = 770.4, [M+H] + .

[1208] Step 10: Preparation of tert-butyl 4-(4-(6-cyano-5-(2-fluoro-6-(4-hydroxybutoxy)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 30-10)

[1209]

[1210] To a solution of intermediate 30-9 (262 mg, 0.34 mmol) in methanol (6 mL) was added p-toluenesulfonic acid monohydrate (65 mg, 0.34 mmol) at room temperature, and the mixture was heated to 40° C. and stirred for 2 hours. The system was rotary evaporated to dryness, and the residue was dissolved in dichloromethane (50 mL), then washed with sodium bicarbonate solution (30 mL), water (30 mL), and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to 1 / 1 to obtain intermediate 30-10 (213 mg, light yellow solid) in a yield of 91.4%.

[1211] ESI-MS: 686.4, [M+H] + .

[1212] Step 11:1 6 -(4-tert-Butyloxycarbonylpiperazin-1-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-2 6 -Cyano-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-indazol-3(1,2)-benzeneheterocyclooctane (Intermediate 30-11)

[1213]

[1214] Under nitrogen, tri-n-butylphosphine (314 mg, 1.55 mmol) was added dropwise to a solution of TMAD (267 mg, 1.55 mmol) in dry tetrahydrofuran (4 mL) at room temperature. After stirring for 5 minutes, dry tetrahydrofuran (20 mL) was added to dilute the mixture, followed by the dropwise addition of a solution of intermediate 30-10 (213 mg, 0.31 mmol) in tetrahydrofuran (5 mL). After the addition was complete, the mixture was stirred at room temperature for 1 hour. Water (80 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to 2 / 1 to obtain intermediate 30-11 (70 mg, white solid) in a 33.8% yield.

[1215] ESI-MS: m / z = 668.4, [M+H] + .

[1216] Step 12:1 6 -(piperazin-1-yl)-2 1 H-2 6 -Cyano-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-indazolyl-3(1,2)-benzylheterocyclooctane hydrochloride (Intermediate 30-12)

[1217]

[1218] To a solution of Intermediate 30-11 (100 mg, 0.14 mmol) in dichloromethane (10 mL) was added dropwise 4M hydrogen chloride in 1,4-dioxane (2 mL) at 0°C, followed by stirring at room temperature for 14 hours. The system was rotary evaporated to dryness to afford Intermediate 30-12 (55 mg, white solid) in a 100% yield.

[1219] ESI-MS: m / z = 484.2, [M+H] + .

[1220] Step 13: Preparation of compound 30

[1221]

[1222] To a solution of intermediate 30-12 (27 mg, 0.06 mmol) in dichloromethane / methanol = 1 / 1 (2 mL) was added 1H-benzotriazole-1-methanol (12 mg, 0.08 mmol), sodium acetate (14 mg, 0.17 mmol) and sodium triacetoxyborohydride (41 mg, 0.20 mmol) at room temperature and stirred at room temperature for 2 hours. The reaction was quenched with sodium bicarbonate solution (20 mL) and extracted with dichloromethane / isopropanol = 5 / 1 (20 mL × 2). The organic phases were combined and washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography eluting with dichloromethane / methanol = 10 / 1 to give the title product (10.5 mg, white solid) in a 38% yield.

[1223] ESI-MS: m / z = 498.3, [M+H] + .

[1224] 1 H NMR(DMSO-d6,400MHz)δ13.73(s,1H),8.30(s,1H),8.10(s,1H)7.44(q,J=8.4Hz,1H),7.24(dd,J=8.0,1.4Hz,1H),7.12(s,1H),7.07-7. 01(m,2H),6.96(t,J=9.2Hz,1H),4.39-4.30(m,1H),4.22-4.07(m,3H),3.20-2.95(m,4H),2.61(s,4H),2.32(s,3H),1.90-1.60(m,4H).

[1225] By selecting appropriate boronate intermediates and using a method similar to Example 30, the following compounds were synthesized:

[1226]

[1227] Example 31

[1228] Compound 31: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 H-2 6 -Cyano-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-indazol-3(1,2)-benzheterocyclooctane

[1229]

[1230] Step 1: Preparation of 5-chloro-3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-6-carbaldehyde (Intermediate 31-1)

[1231]

[1232] To a solution of intermediate 30-4 (800 mg, 2.06 mmol) in dry dichloromethane (15 mL) was added a 1.5 M diisobutylaluminum hydride toluene solution (1.5 mL, 2.2 mmol) dropwise under nitrogen at 0°C, followed by stirring at 0°C for 2 hours. The system was diluted with dichloromethane (10 mL), and aqueous citric acid was added (maintaining pH <4) and stirred overnight. After confirming that the intermediate imine was completely converted to intermediate 31-1, the reaction mixture was washed with dichloromethane (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to afford the crude intermediate 31-1 (800 mg, brown solid) in a 100% yield. The crude product was used directly in the next step.

[1233] ESI-MS: m / z = 412.9, [M+Na] + .

[1234] Step 2: Preparation of 5-chloro-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-hydroxyphenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-6-carbaldehyde (Intermediate 31-2)

[1235]

[1236] Under nitrogen, a mixture of crude Intermediate 31-1 (420 mg, 1.07 mmol), Intermediate A2 (540 mg, 1.08 mmol), Pd(dppf)Cl2 (78 mg, 0.11 mmol), potassium phosphate (684 mg, 3.22 mmol), 1,4-dioxane (10 mL), and water (2 mL) was stirred at 80°C for 4 hours. The temperature was then lowered to 0°C, and a 2M aqueous lithium hydroxide solution (5 mL) was added dropwise, followed by stirring for 1 hour. The pH was adjusted to 7-8 with 5% aqueous citric acid. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution system of petroleum ether / ethyl acetate = 2 / 1 to pure ethyl acetate to afford Intermediate 31-2 (290 mg, brown solid) in a 54% yield.

[1237] ESI-MS: m / z = 497.2, [M+H] + .

[1238] Step 3: Preparation of 5-chloro-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-hydroxypropyloxy)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-6-carbaldehyde (Intermediate 31-3)

[1239]

[1240] Under nitrogen protection, anhydrous potassium carbonate (225 mg, 1.63 mmol) and 3-bromo-1-propanol (90 mg, 0.65 mmol) were added to a solution of intermediate 31-2 (270 mg, 0.54 mmol) in DMF (6 mL), and the mixture was heated to 70°C and stirred for 2 hours. After cooling to room temperature, water (50 mL) was added to the reaction system and extracted with ethyl acetate (50 mL × 2). The organic phases were combined and washed once with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product of intermediate 31-3 (300 mg, yellow oil) was obtained with a yield of 100%. The crude product was not purified and was used directly in the next step.

[1241] ESI-MS: m / z = 555.2, [M+H] + .

[1242] Step 4: Preparation of 5-(2-fluoro-6-hydroxyphenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-hydroxypropyloxy)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-6-carbaldehyde (Intermediate 31-4)

[1243]

[1244] Under nitrogen, a solution of Intermediate 31-3 (300 mg, 0.54 mmol), 2-fluoro-6-hydroxyphenylboronic acid (168 mg, 1.08 mmol), potassium phosphate (344 mg, 1.62 mmol), and Xphos-Pd-G2 (42 mg, 0.050 mmol) in 1,4-dioxane (10 mL) and water (2 mL) was heated to 80°C and stirred for 2 hours. The mixture was cooled to room temperature, diluted with water (60 mL), and extracted with ethyl acetate (50 mL x 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to pure ethyl acetate to obtain Intermediate 31-4 (203 mg, brown solid) in a two-step yield of 59.3%.

[1245] ESI-MS: m / z = 631.3, [M+H] + .

[1246] Step 5: Preparation of 5-(2-fluoro-6-hydroxyphenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-bromopropoxy)phenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-6-carbaldehyde (Intermediate 31-5)

[1247]

[1248] To a solution of intermediate 31-4 (203 mg, 0.32 mmol) in dichloromethane (10 mL) was added triphenylphosphine (110 mg, 0.42 mmol) and NBS (75 mg, 0.42 mmol) at 0°C, followed by stirring at room temperature for 14 hours. After completion of the reaction, the reaction was quenched with sodium bicarbonate solution (30 mL), then extracted with dichloromethane (30 mL × 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to pure ethyl acetate to give the crude intermediate 31-5 (200 mg, pale yellow solid).

[1249] ESI-MS: m / z = 693.2, [M+H] + .

[1250] Step 6: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-2 6 -aldehyde-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-indazolyl-3(1,2)-benzylheterocyclooctane (Intermediate 31-6)

[1251]

[1252] To a solution of intermediate 31-5 (200 mg, 0.288 mmol) in acetonitrile (10 mL) was added cesium carbonate (281 mg, 0.87 mmol) and sodium iodide (22 mg, 0.14 mmol) at room temperature, and the mixture was heated to 70°C and stirred for 2 hours. The mixture was cooled to room temperature, quenched with water (50 mL), and extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of ethyl acetate / methanol = 20 / 1 to give intermediate 31-6 (56 mg, yellow solid) with a two-step yield of 29%.

[1253] ESI-MS: m / z = 613.3, [M+H] + .

[1254] Step 7: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-2 6 -Formaldehyde oxime-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-indazol-3(1,2)-benzeneheterocyclooctane (Intermediate 31-7)

[1255]

[1256] To a solution of intermediate 31-6 (56 mg, 0.09 mmol) in ethanol (3 mL) was added triethylamine (26 μL, 0.19 mmol) and hydroxylamine hydrochloride (13 mg, 0.18 mmol) at room temperature and stirred at room temperature for 3 hours. The mixture was then extracted with ethyl acetate (30 mL × 2), and the organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 31-7 (57 mg, white solid) was obtained in a 100% yield.

[1257] ESI-MS: m / z = 628.3, [M+H] + .

[1258] Step 8: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-2 6 -Cyano-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-indazolyl-3(1,2)-benzylheterocyclooctane (Intermediate 31-8)

[1259]

[1260] To a solution of intermediate 31-7 (57 mg, 0.09 mmol) in DMSO (2 mL) at room temperature were added potassium carbonate (25 mg, 0.18 mmol) and acetic anhydride (17 μL, 0.18 mmol) in sequence. The mixture was heated to 50°C and stirred for 14 hours. The reaction mixture was cooled to room temperature, water (30 mL) was added, and then extracted with ethyl acetate (30 mL x 2). The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. This afforded the crude intermediate 31-8 (75 mg, white solid) in a quantitative yield.

[1261] ESI-MS: m / z = 610.3, [M+H] + .

[1262] Step 9: Synthesis of compound 31

[1263]

[1264] At room temperature, 4M hydrogen chloride in 1,4-dioxane (2 mL) was added dropwise to a solution of intermediate 31-8 (75 mg, 0.09 mmol) in dichloromethane (3 mL) and stirred overnight. The mixture was rotary evaporated to dryness, the pH was adjusted to 9-10 with sodium carbonate solution, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined and washed once with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 12 / 1 as the eluent to give the title product (21 mg, white solid) in a yield of 44.5%.

[1265] ESI-MS: m / z = 526.2, [M+H] + .

[1266] 1 H NMR(DMSO-d6,400MHz)δ13.68(s,1H),8.75(s,1H),8.26(s,1H),7.74(s,1H),7.50-7.45(m,2H),7.08-6.95(m,3H),4.59- 4.39(m,2H),4.14(s,2H),3.82-3.65(m,2H),3.60-3.45(m,2H),3.30-3.12(m,2H),2.82-2.62(m,2H),2.44-2.22(m,7H).

[1267] The following compounds were synthesized using a method similar to Example 31 via appropriate boronate intermediates:

[1268]

[1269]

[1270] Example 33

[1271] Compound 33: (R)-6 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-7- ... 1 H-7 6 -Cyano-8 6 Preparation of 5,9-dioxa-6(1,3)-benzo-7(3,5)-indazol-8(1,2)-benzospiro[2.7]decane

[1272]

[1273] Step 1: Preparation of 5-(2-fluoro-6-hydroxyphenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-hydroxyphenyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-indazole-6-carbaldehyde (Intermediate 33-1)

[1274]

[1275] This compound was prepared according to the procedure described in Example 31 (Step 4) using Intermediate 31-2 instead of Intermediate 31-3 as the starting material. Yield: 78%.

[1276] ESI-MS: m / z = 573.2, [M+H] + .

[1277] Step 2: (R)-6 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-7- ... 1 -(tetrahydro-2H-pyran-2-yl)-7 6 -aldehyde-8 6 Preparation of 5,9-dioxa-6(1,3)-benza-7(3,5)-indazol-8(1,2)-benzaspiro[2.7]decane (Intermediate 33-2)

[1278]

[1279] Intermediate 33-1 (332 mg, 0.580 mmol) was dissolved in acetonitrile (15 mL), and cesium carbonate (567 mg, 1.74 mmol) and 1,1-bis(bromomethyl)cyclopropane (66 mg, 0.29 mmol) were added. The mixture was heated to 70°C and reacted for 4 hours. After completion of the reaction, water (50 mL) was added to quench the reaction. The mixture was then extracted with ethyl acetate (30 mL × 3). The organic phase was collected, dried over anhydrous sodium sulfate, and the solvent was evaporated in vacuo to obtain the crude product. The crude product was purified by preparative thin-layer chromatography using a developing solvent of methanol / ethyl acetate = 1:20. The crude product was purified by reverse-phase medium-pressure preparative chromatography to obtain intermediate 33-2 (8 mg, yellow liquid) in a yield of 2.2%.

[1280] ESI-MS: m / z = 639.3, [M+H] + .

[1281] Compound 33 was prepared according to the preparation method described in Example 31 (steps 7 to 9) using intermediate 33-2 instead of intermediate 31-6 as the starting material. The three-step yield was 20%.

[1282] ESI-MS: m / z = 552.3, [M+H] + .

[1283] 1 H NMR (DMSO-d6, 400 MHz) δ 13.67 (s, 1H), 8.77 (s, 1H), 8.25 (s, 1H), 7.79 (s, 1H), 7.50-7.43 (m, 2H), 7.06-6.96 (m, 3H), 4.68-4.60 (m, 3H), 3.76 (d, J = 10.2 Hz, 1H), 3.67 (t, J = 11.2 Hz, 1H), 3.58-3.33 (m, 4H), 3.26-3.10 (m, 2H), 2.88-2.62 (m, 3H), 2.44-2.22 (m, 4H), 0.68-0.56 (m, 3H), 0.54-0.46 (m, 1H). (monohydrochloride)

[1284] Example 42

[1285] Compound 42:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 6 Preparation of 5,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)iso-3(1,2)-benzheterocyclooctane

[1286]

[1287] Step 1: Preparation of dimethyl 2-(2-(2-bromo-6-fluorophenyl)hydrazono)-3-oxoglutarate (Intermediate 42-1)

[1288]

[1289] To a solution of 2-bromo-6-fluoroaniline (9.5 g, 50.0 mmol) in water (57 mL) at 0°C, 3M hydrochloric acid (90 mL) was added. Then, a solution of sodium nitrite (3.484 g, 50.05 mmol) in water (10 mL) was slowly added dropwise. Stirring was continued for 1 hour, maintaining the reaction temperature below 5°C. After warming to room temperature, the above solution was added dropwise to a mixture of dimethyl 1,3-acetonedicarboxylate (8.794 g, 50.05 mmol) and sodium acetate (24.60 g, 300.0 mmol) in ethanol (57 mL) and water (114 mL). Stirring was continued at room temperature for 1.5 hours. The reaction mixture was filtered, and the filter cake was rinsed with water (50 mL x 2). The filter cake was collected and dried under vacuum to obtain Intermediate 42-1 (18.45 g, yellow solid) in a 98% yield. The crude product was used directly in the next reaction.

[1290] ESI-MS: m / z = 375.1, [M+H] + .

[1291] Step 2: Preparation of methyl 1-(2-bromo-6-fluorophenyl)-4-hydroxy-6-oxo-1,6-dihydropyridazine-3-carboxylate (Intermediate 42-2)

[1292]

[1293] Intermediate 42-1 (18.45 g, 49.20 mmol) was dissolved in 1,2-dichlorobenzene (120 mL) and stirred at 175°C under nitrogen for 7 hours. After the reaction was completed, the temperature was cooled to room temperature and the reaction solution was added dropwise to n-heptane (1200 mL). The solid precipitated with stirring at room temperature and filtered. The filter cake was rinsed with n-heptane (50 mL x 2). The filtrate was separated with saturated sodium bicarbonate solution (1200 mL). The aqueous phase was washed with petroleum ether / ethyl acetate (10 / 1) (200 mL x 2), then adjusted to pH 5 with saturated citric acid solution and extracted with ethyl acetate (200 mL x 3). The organic phase was washed with saturated brine (300 mL x 2), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to dryness. The residue was combined with the filter cake to obtain Intermediate 42-2 (13.57 g) in an 80% yield.

[1294] ESI-MS: m / z = 344.1, [M+H] + .

[1295] Step 3: Preparation of methyl 1-(2-bromo-6-fluorophenyl)-4-chloro-6-oxo-1,6-dihydropyridazine-3-carboxylate (Intermediate 42-3)

[1296]

[1297] Intermediate 42-2 (13.57 g, 39.55 mmol) was dissolved in phosphorus oxychloride (60 mL) and stirred at 100°C overnight under nitrogen. The reaction solution was concentrated under vacuum, and the resulting residue was diluted with dichloromethane (50 mL) and directly purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 20 / 1 to 2 / 1 to obtain intermediate 42-3 (12.40 g, brown solid) in an 86.7% yield.

[1298] ESI-MS: m / z = 361.0, [M+H] + .

[1299] Step 4: Preparation of 1-(2-bromo-6-fluorophenyl)-4-hydrazino-6-oxo-1,6-dihydropyridazine-3-carbohydrazide (Intermediate 42-4)

[1300]

[1301] Intermediate 42-3 (12.40 g, 34.30 mmol) was dissolved in anhydrous ethanol (62 mL). 80% hydrazine hydrate (6.438 g, 102.9 mmol) and DIPEA (17.0 mL, 103 mmol) were added. The mixture was stirred at 80°C under nitrogen for 1.5 hours. The reaction mixture was concentrated under vacuum to afford the crude intermediate 42-4 (12.0 g, brown solid) in a 97.8% yield. This crude product was used directly in the next reaction.

[1302] ESI-MS: m / z = 357.1, [M+H] + .

[1303] Step 5: Preparation of 5-(2-bromo-6-fluorophenyl)-1H-pyrazolo[4,3-c]pyridazine-3,6(2H,5H)-dione (Intermediate 42-5)

[1304]

[1305] The crude intermediate 42-4 (12.0 g, 33.6 mmol) was dissolved in n-butanol (65 mL), and DIPEA (27.8 mL, 168 mmol) and acetic acid (11.5 mL, 202 mmol) were added. The mixture was stirred at 120°C overnight under nitrogen. The reaction solution was concentrated under vacuum, and the residue was purified by column chromatography using a gradient elution from pure dichloromethane to dichloromethane / methanol = 7 / 1 to obtain the crude intermediate 42-5 (10.9 g, brown solid) in a 100% yield. This crude product was used directly in the next reaction.

[1306] ESI-MS: m / z = 325.1, [M+H] + .

[1307] Step 6: Preparation of 5-(2-bromo-6-fluorophenyl)-3-chloro-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 42-6)

[1308]

[1309] The crude intermediate 42-5 (10.90 g) was dissolved in acetonitrile (164 mL), and phosphorus oxychloride (15.6 mL, 168 mmol) and benzyltrimethylammonium chloride (6.227 g, 33.54 mmol) were added. Under nitrogen protection, the mixture was stirred at 70 ° C overnight. The reaction solution was concentrated under vacuum, and the resulting residue was diluted with ice water (200 mL) and extracted with EtOAc (200 mL * 3). The organic phase was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and rotary evaporated to dryness. The residue was slurried with methanol (30 mL), filtered, and the precipitate was collected and dried to obtain intermediate 42-6 (4.20 g, yellow solid) with a yield of 36.5%.

[1310] ESI-MS: m / z = 343.0, [M+H] + .

[1311] Step 7: Preparation of 5-(2-bromo-6-fluorophenyl)-3-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 42-7)

[1312]

[1313] Under nitrogen at 0°C, sodium hydride (768 mg, 19.2 mmol) was added to a solution of intermediate 42-6 (2.20 g, 6.40 mmol) in DMF (25 mL). The mixture was stirred at this temperature for 30 minutes, followed by the dropwise addition of 2-(trimethylsilyl)ethoxymethyl chloride (3.31 g, 19.8 mmol), and the mixture was naturally warmed to room temperature and stirred for 1-2 hours. The mixture was cooled to 0°C, quenched with water (100 mL), and extracted with ethyl acetate (100 mL x 2). The organic phase was washed with water (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 20 / 1 to 5 / 1 as eluent to afford intermediate 42-7 (2.94 g, yellow solid) in a 97% yield.

[1314] ESI-MS: m / z = 475.1, [M+H] + .

[1315] Step 8: Preparation of 3-chloro-5-(2-fluoro-6-vinylphenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 42-8)

[1316]

[1317] Under nitrogen, a mixture of intermediate 42-7 (2.231 g, 4.71 mmol), vinyl pinacol boronate (798 mg, 5.18 mmol), Pd(dppf)Cl2 (516 mg, 0.71 mmol), potassium phosphate (2.99 g, 14.1 mmol), 1,4-dioxane (33 mL), and water (7 mL) was stirred at 100°C for 1 hour. The mixture was then cooled to room temperature, diluted with water (150 mL), and extracted with ethyl acetate (150 mL). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 20 / 1 to 2 / 1 to obtain intermediate 42-8 (1.63 g, light yellow oil) in an 82.3% yield.

[1318] ESI-MS: m / z = 421.2, [M+H] + .

[1319] Step 9: Preparation of 2-(3-chloro-6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3-fluorobenzaldehyde (Intermediate 42-9)

[1320]

[1321] To a solution of intermediate 42-8 (1.63 g, 3.88 mmol), potassium osmate (57 mg, 0.16 mmol), and 2,6-lutidine (829 mg, 27.75 mmol) in 1,4-dioxane (30 mL) and water (10 mL) at 0°C was added sodium periodate (3.32 g, 15.5 mmol) and stirred for 4 hours. The reaction was quenched by the addition of saturated sodium thiosulfate solution (100 mL) and extracted with ethyl acetate (100 mL). The organic phase was washed sequentially with 3% aqueous citric acid (50 mL), water (50 mL), and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to obtain intermediate 42-9 (1.178 g, light yellow oil) in a 72% yield.

[1322] ESI-MS: m / z = 423.2, [M+H] + .

[1323] Step 10: Preparation of 3-chloro-5-(2-fluoro-6-(hydroxymethyl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 42-10)

[1324]

[1325] At 0 ° C and under nitrogen protection, sodium borohydride (106 mg, 2.78 mmol) was added to a solution of intermediate 42-9 (1.178 g, 2.78 mmol) in tetrahydrofuran (30 mL) and stirred at this temperature for 30 minutes. Water (100 mL) was added to quench the reaction and extracted with ethyl acetate (100 mL * 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography with petroleum ether / ethyl acetate = 10 / 1 to 1 / 1 gradient elution to obtain intermediate 42-10 (1.03 g, light yellow oil) in a yield of 86%.

[1326] ESI-MS: m / z = 425.2, [M+H] + .

[1327] Step 11: Preparation of 3-chloro-5-(2-fluoro-6-(bromomethyl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 42-11)

[1328]

[1329] To a solution of intermediate 42-10 (200 mg, 0.47 mmol) in dichloromethane (5 mL) was added triphenylphosphine (246 mg, 0.94 mmol) and carbon tetrabromide (312 mg, 0.94 mmol) at 0°C under nitrogen, and the mixture was stirred overnight at room temperature. The reaction system was evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to afford intermediate 42-11 (223 mg, light yellow oil) in a 97% yield.

[1330] ESI-MS: m / z = 487.2, [M+H] + .

[1331] Step 12: Preparation of 5-(2-((2-((tert-butyldimethylsilyl)oxy)ethoxy)methyl)-6-fluorophenyl)-3-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 42-12)

[1332]

[1333] To a suspension of 60% sodium hydride (49 mg, 1.2 mmol) in dry tetrahydrofuran (5 mL) at 0°C under nitrogen was added tert-butyldimethylhydroxyethoxysilane (410 mg, 2.46 mmol). The mixture was stirred in an ice-water bath for 0.5 hours and then stirred at room temperature for 2 hours. A solution of intermediate 42-11 (200 mg, 0.41 mmol) in tetrahydrofuran (3 mL) was then slowly added dropwise and stirred at room temperature overnight. The reaction was quenched by adding water (100 mL) and extracted with ethyl acetate (100 mL*2). The organic phase was washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 2 / 1 to obtain intermediate 42-12 (85 mg, light yellow oil) in a 40% yield.

[1334] ESI-MS: m / z = 583.4, [M+H] + .

[1335] Step 13: Preparation of tert-butyl 4-(4-(5-(2-((2-((tert-butyldimethylsilyl)oxy)ethoxy)methyl)-6-fluorophenyl)-6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-5,6-dihydro-1H-pyrazolo[4,3-c]pyridazin-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 42-13)

[1336]

[1337] Under nitrogen, a solution of Intermediate 42-12 (85 mg, 0.16 mmol), Intermediate A3 (162 mg, 0.36 mmol), potassium phosphate (93 mg, 0.44 mmol), and Sphos-Pd-G2 (11 mg, 0.015 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was heated to 100°C and stirred overnight. The mixture was cooled to room temperature, diluted with water (40 mL), and extracted with ethyl acetate (40 mL × 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was then dissolved in methanol (5 mL), cooled to 0°C, and 3M lithium hydroxide solution (1 mL) was added dropwise and stirred for 30 minutes. The pH was adjusted to 7 with 5% citric acid, followed by addition of water (30 mL) and extraction with ethyl acetate (40 mL × 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to 2 / 1 to afford intermediate 42-13 (93 mg, yellow oil) in a 63% yield.

[1338] ESI-MS: m / z = 825.7, [M+H] + .

[1339] Step 14: Preparation of tert-butyl 4-(4-(5-(2-((2-hydroxyethoxy)methyl)-6-fluorophenyl)-6-oxo-1-((2-(trimethylsilyl)ethoxy)methyl)-5,6-dihydro-1H-pyrazolo[4,3-c]pyridazin-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 42-14)

[1340]

[1341] At 0 ° C, to a solution of intermediate 42-13 (93 mg, 0.11 mmol) in tetrahydrofuran (5 mL) was added a 1M TBAF tetrahydrofuran solution (0.12 mL, 0.12 mmol) and stirred at room temperature overnight. Water (50 mL) was added to quench the reaction and extracted with ethyl acetate (50 mL * 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 1 / 1 to pure ethyl acetate to give intermediate 42-14 (62 mg, yellow solid) in a yield of 77.5%.

[1342] ESI-MS: m / z = 711.6, [M+H] + .

[1343] Step 15:16 -(4-tert-Butyloxycarbonylpiperazin-1-yl)-2 1 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 6 Preparation of 5,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclooctane (Intermediate 42-15)

[1344]

[1345] Under nitrogen protection, tri-n-butylphosphine (70 mg, 0.35 mmol) was added dropwise to a solution of TMAD (60 mg, 0.35 mmol) in dry tetrahydrofuran (10 mL) at room temperature. After stirring for 5 minutes, a solution of intermediate 42-14 (62 mg, 0.09 mmol) in tetrahydrofuran (3 mL) was added dropwise. After the addition was complete, the mixture was stirred at room temperature for 2 hours. Water (50 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to 1 / 1 to obtain intermediate 42-15 (30 mg, yellow oil) in a 50% yield.

[1346] ESI-MS: m / z = 693.5, [M+H] + .

[1347] Step 16:1 6 -(piperazin-1-yl)-2 1 H-3 6 Preparation of 5,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclooctane (Intermediate 42-16)

[1348]

[1349] To a solution of intermediate 42-15 (30 mg, 0.040 mmol) in dichloromethane (5 mL) at 0°C was added trifluoroacetic acid (1.0 mL) dropwise, followed by stirring at room temperature for 14 hours. The system was rotary evaporated to dryness, redissolved in dichloromethane (20 mL), and sodium carbonate solution (20 mL) was added. The product was extracted with dichloromethane / isopropanol = 4 / 1 (20 mL x 3). The extracts were combined and rotary evaporated to dryness to obtain the crude product. The crude product was redissolved in acetonitrile (5 mL), and ammonia water (0.5 mL) was added and stirred for 30 minutes. Water (30 mL) was added, and the product was extracted with dichloromethane / isopropanol = 4 / 1 (20 mL x 3). The organic phases were combined and washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 42-16 (23 mg, yellow solid) was obtained in a 100% yield.

[1350] ESI-MS: m / z = 463.3, [M+H] + .

[1351] Step 17: Preparation of compound 42

[1352]

[1353] To a solution of intermediate 42-16 (23 mg, 0.050 mmol) in dichloromethane / methanol = 1 / 1 (4 mL) was added 1H-benzotriazole-1-methanol (10 mg, 0.060 mmol), sodium acetate (10 mg, 0.13 mmol) and sodium triacetoxyborohydride (26 mg, 0.13 mmol) at room temperature and stirred at room temperature for 2 hours. The reaction was quenched with sodium bicarbonate solution (30 mL) and extracted with dichloromethane / isopropanol = 4 / 1 (20 mL × 3). The organic phases were combined and washed once with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography eluting with dichloromethane / methanol = 6 / 1 to give the title product (8.4 mg, orange solid) in a 35.6% yield.

[1354] ESI-MS: m / z = 477.2, [M+H] + .

[1355] 1H NMR(DMSO-d6,400MHz)δ8.56(s,1H),7.63-7.50(m,3H),7.45-7.35(m,1H),6.98(d,J=8.8Hz,1H),6.64(s,1H),4.55(d,J =10.4Hz,1H),4.39(d,J=10.0Hz,1H),4.22(s,2H),3.83-3.59(m,4H),3.00-2.90(m,2H),2.50-2.40(m,4H),2.21(s,3H).

[1356] Example 43

[1357] Compound 43:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 6 Preparation of 5,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane

[1358]

[1359] Step 1: Preparation of N-(diphenylmethylene)-2-fluoro-6-((3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)methyl)aniline (Intermediate 43-1)

[1360]

[1361] Under nitrogen, a toluene (50 mL) solution of 2-(3-((2-bromo-3-fluorobenzyl)oxy)propoxy)tetrahydro-2H-pyran (4.000 g, 11.52 mmol), benzophenone imine (3.132 g, 17.28 mmol), Pd2(dba)3 (527 mg, 0.58 mmol), BINAP (717 mg, 1.15 mmol) and sodium tert-butoxide (2.768 g, 28.80 mmol) was heated to 100°C and stirred for 14 hours. The mixture was cooled to room temperature, water (100 mL) was added to the reaction system, and the mixture was extracted with ethyl acetate (80 mL × 2). The organic phases were combined and washed once with water (100 mL) and saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 20 / 1 to give intermediate 43-1 (4.8 g, yellow oil) in a 93% yield.

[1362] ESI-MS: m / z = 448.2, [M+H] + .

[1363] Step 2: Preparation of 3-((2-amino-3-fluorobenzyl)oxy)propan-1-ol (Intermediate 43-2)

[1364]

[1365] At 0°C, a 4M solution of hydrogen chloride in 1,4-dioxane (20 mL) was added dropwise to a solution of intermediate 43-1 (4.80 g, 10.7 mmol) in dichloromethane (15 mL) and stirred for 30 minutes. Water (10 mL) was then added and stirred at room temperature for 1 hour. The pH was adjusted to 9 with sodium carbonate solution, extracted with dichloromethane (50 mL × 2), and the organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 1 / 1 as the eluent to give intermediate 43-2 (1.616 g, white solid) in a 68% yield.

[1366] ESI-MS: m / z = 222.1, [M+Na] + .

[1367] Step 3: Preparation of 2-fluoro-6-((3-hydroxypropoxy)methyl)phenyldiazonium tetrafluoroborate (Intermediate 43-3)

[1368]

[1369] To a solution of intermediate 43-2 (1.30 g, 6.53 mmol) in tetrahydrofuran (17 mL) was added dropwise 48% (w / w) tetrafluoroboric acid aqueous solution (2.39 g, 13.1 mmol) under nitrogen protection at 0°C. After stirring for 30 minutes, tert-butyl nitrite (1.347 g, 13.07 mmol) was added dropwise. The mixture was stirred at this temperature for 40 minutes and evaporated to dryness at low temperature to give intermediate 43-3 (1.94 g, light yellow oil) with a yield of 100%.

[1370] Step 4: Preparation of 4-bromo-3-hydroxybenzaldehyde (Intermediate 43-4)

[1371]

[1372] Under nitrogen protection, boron tribromide (6.3 mL, 68 mmol) was added dropwise to a solution of 4-bromo-3-methoxybenzaldehyde (4.908 g, 22.83 mmol) in dichloromethane (50 mL) at -70°C, and the mixture was then allowed to rise to room temperature and stirred for 2 hours. The temperature was lowered to -20°C, and methanol (10 mL) was slowly added dropwise to quench the reaction, followed by rotary evaporation to dryness. The crude product was dissolved in a mixture of methanol and dichloromethane, to which 2 M sodium hydroxide solution was added dropwise (maintaining pH > 12) and stirred for 30 minutes. The pH was then adjusted to 3 with dilute hydrochloric acid, and the mixture was extracted with dichloromethane (100 mL x 2). The organic phase was washed once with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Intermediate 43-4 (3.1 g) was obtained with a yield of 67%.

[1373] Step 5: Preparation of 4-bromo-3-((tert-butyldimethylsilyl)oxy)benzaldehyde (Intermediate 43-5)

[1374]

[1375] At room temperature, imidazole (3.097 g, 30.84 mmol) and tert-butyldimethylsilyl chloride (3.486 g, 23.15 mmol) were added to a solution of intermediate 43-4 (3.100 g, 15.42 mmol) in dichloromethane (30 mL), and the mixture was stirred at room temperature for 30 minutes. The mixture was quenched with aqueous sodium bicarbonate solution and extracted with dichloromethane (50 mL × 2). The organic phase was washed once with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 20 / 1 to give intermediate 43-5 (3.426 g, brown oil) in a yield of 70.5%.

[1376] Step 6: Preparation of 6-(2-(4-bromo-3-((tert-butyldimethylsilyl)oxy)phenyl)-2-hydroxyethyl)-2,2-dimethyl-4H-1,3-dioxin-4-one (Intermediate 43-6)

[1377]

[1378] To a solution of 2,2,6-trimethyl-4H-1,3-dioxin-4-one (1.700 g, 11.95 mmol) in tetrahydrofuran (20 mL) was added dropwise a 1 M solution of LiHMDS in tetrahydrofuran (12 mL, 12 mmol) at -70°C under nitrogen, maintaining the internal temperature below -60°C. The mixture was then stirred at -65°C for 30 minutes, followed by the dropwise addition of a solution of Intermediate 43-5 (3.426 g, 10.87 mmol) in tetrahydrofuran (20 mL). After the addition was complete, the mixture was stirred for 0.5 hours. After completion of the reaction, the reaction was quenched by the addition of saturated ammonium chloride (100 mL) at below -20°C, gradually heated to 0°C, diluted with water (50 mL), and extracted with ethyl acetate (80 mL of acetic acid). The organic phase was washed once with water (80 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient of petroleum ether / ethyl acetate = 20 / 1 to 3 / 1 to afford Intermediate 43-6 (4.075 g, brown oil) in a 75% yield.

[1379] ESI-MS: m / z = 457.2, [M+H] + .

[1380] Step 7: Preparation of 6-(2-(4-bromo-3-((tert-butyldimethylsilyl)oxy)phenyl)-2-oxoethyl)-2,2-dimethyl-4H-1,3-dioxin-4-one (Intermediate 43-7)

[1381]

[1382] At 0.3 °C, intermediate 43-6 (4.075 g, 8.91 mmol) was dissolved in dichloromethane (40 mL).

[1383] Dess-Martin periodinane (4.930 g, 11.58 mmol) was added to the mixture, followed by stirring at room temperature for 1 hour. After completion of the reaction, the temperature was lowered to 0°C and quenched with saturated aqueous sodium thiosulfate (30 mL). The pH was adjusted to 7-8 with saturated sodium bicarbonate. The mixture was separated, and the aqueous phase was extracted with dichloromethane (50 mL). The organic phases were combined, washed with water (50 mL), washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 20 / 1 to 5 / 1 to obtain intermediate 43-7 (3.49 g, yellow oil) in an 86% yield.

[1384] Step 8: Preparation of 6-(2-(4-bromo-3-hydroxyphenyl)-1-(2-(2-fluoro-6-((3-hydroxypropyloxy)methyl)phenyl)hydrazono)-2-oxoethyl)-2,2-dimethyl-4H-1,3-dioxin-4-one (Intermediate 43-8)

[1385]

[1386] At room temperature, intermediate 43-7 (400 mg, 0.90 mmol) was dissolved in anhydrous ethanol (4 mL).

[1387] Anhydrous sodium acetate (252 mg, 0.31 mmol) was added to the mixture, followed by a solution of intermediate 43-3 (300 mg, 1.00 mmol) in acetonitrile (4 mL) dropwise. The mixture was allowed to react at room temperature for 0.5 h. After completion of the addition, the reaction was quenched with saturated ammonium chloride and extracted with ethyl acetate (30 mL). The organic phase was washed once with water (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 1 / 1 to obtain intermediate 43-8 (302 mg, yellow oil) in a 62% yield.

[1388] Step 9: Preparation of 6-(2-(4-bromo-3-((tert-butyldimethylsilyl)oxy)phenyl)-1-(2-(2-((3-((tert-butyldimethylsilyl)oxy)propoxy)methyl)-6-fluorophenyl)hydrazono)-2-oxoethyl)-2,2-dimethyl-4H-1,3-dioxin-4-one (Intermediate 43-9)

[1389]

[1390] To a solution of intermediate 43-8 (245 mg, 0.45 mmol) in dichloromethane (8 mL) was added imidazole (76 mg, 1.1 mmol) and tert-butyldimethylsilyl chloride (141 mg, 0.93 mmol) at room temperature, followed by stirring at room temperature for 30 minutes. The mixture was quenched with water (30 mL) and extracted with dichloromethane (30 mL of dichloromethane). The organic phase was washed once with water (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 10:1 as eluent to afford intermediate 43-9 (184 mg, yellow oil) in a 53% yield.

[1391] ESI-MS: m / z = 779.3, [M+H] + .

[1392] Step 10: Preparation of 6-(4-bromo-3-((tert-butyldimethylsilyl)oxy)benzoyl)-2-(2-((3-((tert-butyldimethylsilyl)oxy)propoxy)methyl)-6-fluorophenyl)-5-hydroxypyridazin-3(2H)-one (Intermediate 43-10)

[1393]

[1394] At room temperature, intermediate 43-9 (184 mg, 0.24 mmol) was dissolved in o-dichlorobenzene (2 mL).

[1395] Glacial acetic acid (92 μL, 0.5 V) was added to the mixture, which was then heated to 130°C and stirred in an oil bath for 1 hour. After the reaction was completed, the mixture was cooled to room temperature and directly purified by column chromatography using dichloromethane / methanol = 10 / 1 as the eluent to afford intermediate 43-10 (155 mg, light yellow oil) in a 70% yield.

[1396] ESI-MS: m / z = 721.3, [M+H] + .

[1397] Step 11: Preparation of 3-(4-bromo-3-((tert-butyldimethylsilyl)oxy)phenyl)-5-(2-((3-((tert-butyldimethylsilyl)oxy)propoxy)methyl)-6-fluorophenyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 43-11)

[1398]

[1399] Under nitrogen protection, 80% hydrazine hydrate (65 μL, 1.1 mmol) and glacial acetic acid (61 μL, 1.1 mmol) were added to a solution of intermediate 43-10 (184 mg, 0.24 mmol) in 2-methyl-2-butanol (2 mL) at room temperature, and then the temperature was raised to 100°C and stirred for 1 hour. After the reaction was completed, the mixture was cooled to room temperature, water (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL of ethyl acetate). The organic phase was washed once with water (30 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 10 / 1 to 1 / 1 to obtain intermediate 43-11 (100 mg, yellow solid) in a yield of 65%.

[1400] ESI-MS: m / z = 717.2, [M+H] + .

[1401] Step 12: Preparation of 3-(4-bromo-3-hydroxyphenyl)-5-(2-((3-((tert-butyldimethylsilyl)oxy)propoxy)methyl)-6-fluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 43-12)

[1402]

[1403] Under the protection of 03 and nitrogen, anhydrous potassium carbonate (58mg, 0.42mmol) was added to a DMF (2mL) solution of intermediate 43-11 (100mg, 0.14mmol), followed by dropwise addition of 2-(trimethylsilyl)ethoxychloromethane (32μL, 0.18mmol), and the mixture was naturally warmed to room temperature and stirred overnight. The reaction was quenched with water (30mL) and extracted with ethyl acetate (30mL, and), the organic phase was washed with water (30mL), saturated brine (30mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography, eluted with petroleum ether / ethyl acetate = 10 / 1 to 2 / 1, to give intermediate 43-12 (63mg, yellow solid) in a yield of 61.7%.

[1404] ESI-MS: m / z = 733.2, [M+H] + .

[1405] Step 13: Preparation of 3-(4-bromo-3-hydroxyphenyl)-5-(2-fluoro-6-((3-hydroxypropyloxy)methyl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 43-13)

[1406]

[1407] 03 and nitrogen protection, to the tetrahydrofuran solution of intermediate 43-12 (63 mg, 0.09 mmol) was added 1M TBAF tetrahydrofuran solution (94 μL, 0.10 mmol), then warmed to room temperature and stirred overnight. The reaction was quenched with water (30 mL) and extracted with ethyl acetate (30 mL, and), the organic phase was washed with water (30 mL), saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography, eluting with petroleum ether / ethyl acetate = 5 / 1 to pure ethyl acetate to obtain intermediate 43-13 (60 mg, yellow oil) in a yield of 113%.

[1408] ESI-MS: m / z = 619.2, [M+H] + .

[1409] Step 14:1 6 -Br-2 1 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 6 Preparation of 5,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane (Intermediate 43-14)

[1410]

[1411] Under nitrogen protection, tri-n-butylphosphine (78 mg, 0.39 mmol) was added dropwise to a solution of TMAD (67 mg, 0.39 mmol) in dry tetrahydrofuran (5 mL) at room temperature. After stirring for 5 minutes, a solution of intermediate 43-13 (60 mg, 0.09 mmol) in tetrahydrofuran (2 mL) was added dropwise. After completion of the addition, the mixture was stirred at room temperature for 1 hour. Water (30 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (30 mL of acetic acid). The organic phase was washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using a gradient elution of petroleum ether / ethyl acetate = 5 / 1 to 1 / 1 to obtain intermediate 43-14 (25 mg, white solid) in a 42% yield.

[1412] ESI-MS: m / z = 601.2, [M+H] + .

[1413] Step 15:1 6 -(4-methylpiperazin-1-yl)-2 1 -((2-(trimethylsilyl)ethoxy)methyl)-3- ... 6 Preparation of 5,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane (Intermediate 43-15)

[1414]

[1415] Under nitrogen, a solution of intermediate 43-14 (25 mg, 0.04 mmol), 1-methylpiperazine (17 mg, 0.16 mmol), palladium acetate (1 mg, 0.004 mmol), BINAP (5 mg, 0.008 mmol), and cesium carbonate (41 mg, 0.12 mmol) in toluene (1 mL) was heated to 100 liters and stirred for 14 hours. The mixture was cooled to room temperature, and water (10 mL) was added to the reaction system. The mixture was extracted with ethyl acetate (10 mL). The organic phases were combined and washed once with water (10 mL) and once with saturated brine (10 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 15 / 1 as the eluent to afford intermediate 43-15 (7 mg, yellow oil) in a 34% yield.

[1416] ESI-MS: m / z = 621.3, [M+H] + .

[1417] Step 16: Preparation of compound 43

[1418]

[1419] To a solution of intermediate 43-15 (7 mg, 0.01 mmol) in dichloromethane (1 mL) was added trifluoroacetic acid (0.25 mL) dropwise at 0°C, followed by stirring at room temperature for 5 hours. The system was rotary evaporated to dryness, redissolved in acetonitrile (1 mL), to which was added aqueous ammonia (0.1 mL) and stirred for 30 minutes. Evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 10 / 1 as the eluent to give the title product (2.1 mg, orange solid) in a 38% yield.

[1420] ESI-MS: m / z = 491.2, [M+H] + .

[1421] 1 H NMR (DMSO-d6, 400MHz), δ12.88 (s, 1H), 8.12 (d, J = 7.6Hz, 1H), 7.63-7.56 (m, 1H),7.54-7.48(m,2H),7.46-7.40(m,1H),6.99(d,J=8.4Hz,1H),6.65(s,1H) 4.46(d,J=11.6Hz,1H),4.28-4.20(m,2H),4.16-4.08(m,1H),3.56(d,J=4.8H z,2H),3.10-2.90(m,4H),2.50-2.40(m,4H),2.22(s,3H),2.05-1.75(m,2H).

[1422] Example 44

[1423] Compound 44: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 H-3 6 Preparation of 4,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane

[1424]

[1425] Step 1: Preparation of 3-chloro-5-(2-fluoro-6-methoxyphenyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-1)

[1426]

[1427] This compound was prepared according to the preparation method of intermediate 42-6 described in Example 42 (steps 1 to 6) using 2-methoxy-6-fluoroaniline instead of 2-bromo-6-fluoroaniline as the starting material.

[1428] ESI-MS: m / z = 294.2, [M+H] + .

[1429] Step 2: Preparation of 3-chloro-5-(2-fluoro-6-hydroxyphenyl)-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-2)

[1430]

[1431] A solution of intermediate 44-1 (1804 mg, 6.12 mmol) in dichloromethane (38 mL) was cooled.

[1432] To 0 liquid, then slowly add boron tribromide (1.70 mL, 18.4 mmol), then warm to room temperature and stir for 3 hours. After the reaction is completed, cool to 0 again, then quench with methanol (5 mL), concentrate under reduced pressure, dry in vacuum, and slurry with methanol to obtain intermediate 44-2 (1.615 g, yellow solid) with a yield of 94%.

[1433] ESI-MS: m / z = 281.0, [M+H] + .

[1434] Step 3: Preparation of 5-(2-((tert-butyldimethylsilyl)oxy)-6-fluorophenyl)-3-chloro-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-3)

[1435]

[1436] To a solution of intermediate 44-2 (1.465 g, 5.22 mmol) in tetrahydrofuran (20 mL) was added DIPEA (2.728 mL, 15.66 mmol), stirred at room temperature for 3 minutes, then added tert-butyldimethylsilyl chloride (958 mg, 6.36 mmol), and stirred at room temperature for 7 hours. After completion of the reaction, the mixture was quenched with water (150 mL), cooled to room temperature, and extracted with ethyl acetate (50 mL to room temperature). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether / ethyl acetate = 5 / 1 to 2 / 1 as eluent to obtain intermediate 44-3 (2.114 g, yellow solid) in a yield of 103%.

[1437] ESI-MS: m / z = 395.1, [M+H] + .

[1438] Step 4: Preparation of 5-(2-((tert-butyldimethylsilyl)oxy)-6-fluorophenyl)-3-chloro-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-4)

[1439]

[1440] To a solution of intermediate 44-3 (2.114 g, 5.353 mmol) in DMF (25 mL) was added

[1441] DIPEA (1.40mL, 8.03mmol), stirred at normal temperature for 3 minutes, then triphenylmethane (1.713g, 6.16mmol) was added, and then stirred at room temperature for 3 hours. After the reaction was completed, it was quenched with water (150mL) and extracted with ethyl acetate (50mL with second solvent). The organic phase was washed with saturated aqueous common salt (150mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by chromatography using petroleum ether / ethyl acetate = 10 / 1 as eluent to obtain intermediate 44-4 (2.996g, yellow solid) in a yield of 88%.

[1442] ESI-MS: m / z = 637.2, [M+H] + .

[1443] Step 5: Preparation of 3-chloro-5-(2-fluoro-6-hydroxyphenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-5)

[1444]

[1445] To a solution of intermediate 44-4 (2.996 g, 4.702 mmol) in tetrahydrofuran (60 mL) was added 1 M

[1446] A solution of tetrabutylammonium fluoride in tetrahydrofuran (5 mL) was stirred at room temperature for 1.5 hours. After completion of the reaction, the mixture was quenched with saturated ammonium chloride solution (300 mL) and extracted with dichloromethane / methanol = 10 / 1 (100 mL dichloromethane). The organic phase was washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was crystallized with methanol and slurried to give intermediate 44-5 (2.308 g, yellow solid) in a 94% yield.

[1447] ESI-MS: m / z = 523.1, [M+H] + .

[1448] Step 6: Preparation of 3-chloro-5-(2-fluoro-6-(4-((tetrahydro-2H-pyran-2-yl)oxy)butoxy)phenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-6)

[1449]

[1450] To a solution of intermediate 44-5 (601 mg, 1.15 mmol) in DMF (10 mL) were added potassium carbonate (300 mg, 2.30 mmol) and 2-(4-bromobutoxy)tetrahydro-2H-pyran (364 mg, 1.54 mmol), and the mixture was stirred at 80°C for 14 hours. After completion of the reaction, the mixture was quenched with water (150 mL) and extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (150 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using petroleum ether:ethyl acetate = 5 / 1 to 2 / 1 as eluent to obtain intermediate 44-6 (809 mg, yellow solid) in a yield of 104%.

[1451] ESI-MS: m / z = 679.2, [M+H] + .

[1452] Step 7: Preparation of 5-(2-fluoro-6-(4-((tetrahydro-2H-pyran-2-yl)oxy)butoxy)phenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-hydroxyphenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-7)

[1453]

[1454] To a solution of Intermediate 44-6 (377 mg, 0.555 mmol) in 1,4-dioxane (8 mL) and water (2 mL) were added Intermediate A2 (383 mg, 1.063 mmol), Xphos-Pd-G2 (38 mg, 0.053 mmol), and potassium phosphate (342 mg, 1.611 mmol). The mixture was then heated to 100 L and stirred for 14 hours. After completion of the reaction, the mixture was cooled to room temperature and stirred at room temperature for 1 hour. The mixture was then adjusted to neutral with aqueous citric acid and extracted with ethyl acetate (30 mL). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using dichloromethane / methanol = 30 / 1 as the eluent to afford Intermediate 44-7 (409 mg, red solid) in an 84% yield.

[1455] ESI-MS: m / z = 877.4, [M+H] + .

[1456] Step 8: Preparation of 5-(2-fluoro-6-(4-hydroxybutoxy)phenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-hydroxyphenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-8)

[1457]

[1458] To a solution of intermediate 44-7 (409 mg, 1.47 mmol) in methanol (10 mL) was added p-toluenesulfonic acid pyridinium salt (123 mg, 0.489 mmol), the temperature was raised to 50 liters and stirred for 14 hours. If the reaction was not complete, heating was stopped, the temperature was lowered to room temperature, p-toluenesulfonic acid monohydrate (88 mg) was added, the temperature was raised to 40 liters, and after stirring for 1 hour, the reaction was completed and quenched with sodium bicarbonate aqueous solution (50 mL) and extracted with dichloromethane (30 mL with dichloromethane). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by column chromatography using dichloromethane / methanol = 50 / 1 to 30 / 1 as eluent to obtain intermediate 44-8 (330 mg, red solid) in a yield of 89%.

[1459] ESI-MS: m / z = 793.3, [M+H] + .

[1460] Step 9: Preparation of 5-(2-fluoro-6-(4-bromobutoxy)phenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-hydroxyphenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 44-9)

[1461]

[1462] To a solution of intermediate 44-8 (80 mg, 0.101 mmol) in dichloromethane (3 mL) was added triphenylphosphine (33 mg, 0.17 mmol), cooled to 0°C with stirring, and NBS (22 mg, 0.12 mmol) was added. The reaction was allowed to return to room temperature for 14 hours. After the reaction was completed, the mixture was quenched with aqueous sodium bicarbonate (50 mL) and extracted with dichloromethane (30 mL with dichloromethane). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 17 / 1 as the developing solvent to obtain intermediate 44-9 (64 mg, red solid) in a yield of 74%.

[1463] ESI-MS: m / z = 855.3, [M+H] + .

[1464] Step 10: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 -Trityl-3 6 Preparation of 4,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane (Intermediate 44-10)

[1465]

[1466] To a solution of intermediate 44-9 (64 mg, 0.075 mmol) in acetonitrile (3 mL) was added cesium carbonate (74 mg, 0.23 mmol) and sodium iodide (8 mg, 0.053 mmol), the temperature was raised to 70 ° C and stirred for 1 hour. The reaction was completed, quenched with an aqueous solution (50 mL), and extracted with dichloromethane (30 mL with dichloromethane). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 17 / 1 as the developing solvent to obtain intermediate 44-10 (41 mg, red solid) in a yield of 72%.

[1467] ESI-MS: m / z = 775.3, [M+H] + .

[1468] Step 11: Preparation of compound 44

[1469]

[1470] To a solution of intermediate 44-10 (41 mg) in dichloromethane (3 mL) was added trifluoroacetic acid (1 mL), stirred at room temperature for 14 hours, the reaction was completed, quenched with saturated sodium bicarbonate aqueous solution (50 mL), and extracted with dichloromethane (30 mL with dichloromethane), the organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 11 / 1 as the developing solvent, and the crude product was slurried with dichloromethane and n-heptane to give the title product (4 mg, red solid).

[1471] ESI-MS: m / z = 533.2, [M+H] + .

[1472] Example 45

[1473] Compound 45:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 6 Preparation of 4,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane

[1474]

[1475] Step 1: Preparation of tert-butyl 4-(4-(5-(2-fluoro-6-(4-((tetrahydro-2H-pyran-2-yl)oxy)butoxy)phenyl)-6-oxo-1-trityl-5,6-dihydro-1H-pyrazolo[4,3-c]pyridazin-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 45-7)

[1476]

[1477] This compound was prepared according to the procedure described in Example 44 (step 7) using Intermediate A3 instead of Intermediate A2 as the starting material.

[1478] ESI-MS: m / z = 920.5, [M+H] + .

[1479] Step 2: Preparation of tert-butyl 4-(4-(5-(2-fluoro-6-(4-hydroxybutoxy)phenyl)-6-oxo-1-trityl-5,6-dihydro-1H-pyrazolo[4,3-c]pyridazin-3-yl)-2-hydroxyphenyl)piperazine-1-carboxylate (Intermediate 45-8)

[1480]

[1481] This compound was prepared according to the procedure described in Example 44 (Step 8) using Intermediate 45-7 instead of Intermediate 44-7 as the starting material. Yield: quantitative.

[1482] ESI-MS: m / z = 837.4, [M+H] + .

[1483] Step 3:1 6 -(4-(tert-Butyloxycarbonyl)piperazin-1-yl)-2 1 -Trityl-3 6 Preparation of 4,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane (Intermediate 45-9)

[1484]

[1485] To a solution of TMAD (88 mg, 0.514 mmol) in tetrahydrofuran (1 mL) was added tri-n-butylphosphine (104 mg, 0.514 mmol), stirred at room temperature for 10 minutes under nitrogen protection, then diluted with tetrahydrofuran (5 mL), a solution of intermediate 45-8 in tetrahydrofuran (4 mL) was added, and the temperature was raised to 50°C and stirred for 30 minutes. After the reaction was completed, it was quenched with water (50 mL) and cooled to room temperature, and extracted with dichloromethane (30 mL to room temperature). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative thin layer chromatography using dichloromethane / methanol = 15 / 1 as the developing solvent to obtain intermediate 45-9 (17 mg, yellow solid) with a two-step yield of 22%.

[1486] ESI-MS: m / z = 819.4, [M+H] + .

[1487] Step 4:1 6 -(piperazin-1-yl)-2 1 H-3 6 Preparation of 4,9-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzoheterocyclodecane (Intermediate 45-10)

[1488]

[1489] To a solution of intermediate 45-9 (17 mg, 0.036 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (150 mL) and stirred at room temperature for 16 hours. The reaction mixture was detected by liquid chromatography-mass spectrometry to determine the residual intermediate. Trifluoroacetic acid (0.5 mL) was added and the reaction was continued at room temperature for 3 hours. After completion of the reaction, the mixture was quenched with sodium bicarbonate solution (50 mL) and extracted with dichloromethane (30 mL with dichloromethane). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative thin layer chromatography using dichloromethane / methanol = 5 / 1 as the developing solvent to obtain intermediate 45-10 (9 mg, yellow solid) in a yield of 91%.

[1490] ESI-MS: m / z = 477.2, [M+H] + .

[1491] Step 5: Preparation of compound 45

[1492]

[1493] To a solution of intermediate 45-10 (9 mg, 0.019 mmol) in dichloromethane (2 mL) was added methanol (2 mL), sodium acetate (5 mg, 0.06 mmol), sodium triacetoxyborohydride (12 mg, 0.057 mmol) and 1H-benzotriazole-1-methanol (4 mg, 0.03 mmol) and stirred at room temperature for 1 hour. After completion of the reaction, the mixture was quenched with sodium bicarbonate solution (50 mL) and extracted with dichloromethane (30 mL with dichloromethane). The organic phase was washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The residue was purified by preparative chromatography using dichloromethane / methanol = 7 / 1 as the developing solvent. The crude product was crystallized and slurried with dichloromethane and n-heptane to give the title product (3 mg, orange solid) in a 32% yield.

[1494] ESI-MS: m / z = 491.2, [M+H] + .

[1495] 1H NMR(DMSO-d6,400MHz)δ12.83(s,1H),7.76(d,J=1.9Hz,1H),7.47(td,J=8.6,6.6Hz,1H),7.39( dd,J=8.1,1.8Hz,1H),7.16(d,J=8.7Hz,1H),7.00(d,J=8.3Hz,1H),6.95(t,J=8.9Hz,1H),6.66( s,1H),4.50-4.44(m,1H),4.29-4.13(m,3H),3.16-3.06(m,2H),3.04-2.90(m,2H),2.60-2.50( m,2H),2.26(s,3H),2.18-2.06(m,1H),2.06-1.94(m,1H),1.83-1.71(m,1H),1.70-1.60(m,1H).

[1496] Example 46

[1497] Compound 46:1 6 -(4-methylpiperazin-1-yl)-2 1 H-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclooctane

[1498]

[1499] Step 1: Preparation of tert-butyl 4-(4-(5-(2-fluoro-6-hydroxyphenyl)-6-oxo-1-trityl-5,6-dihydro-1H-pyrazolo[4,3-c]pyridazin-3-yl)-2-(3-hydroxypropyloxy)phenyl)piperazine-1-carboxylate (Intermediate 46-7)

[1500]

[1501] This compound was prepared according to the procedure described in Example 44 (steps 7 to 8) using Intermediate A5 instead of Intermediate A2 as the starting material. Yield: 93%.

[1502] ESI-MS: m / z = 823.5, [M+H] + .

[1503] Step 2: Preparation of tert-butyl 4-(4-(5-(2-fluoro-6-hydroxyphenyl)-6-oxo-1-trityl-5,6-dihydro-1H-pyrazolo[4,3-c]pyridazin-3-yl)-2-(3-bromopropoxy)phenyl)piperazine-1-carboxylate (Intermediate 46-8)

[1504]

[1505] This compound was prepared according to the procedure described in Example 44 (Step 9) using Intermediate 46-7 instead of Intermediate 44-8 as the starting material. Yield: 58%.

[1506] ESI-MS: m / z = 885.5, [M+H] + .

[1507] Step 3:1 6 -(4-(tert-Butyloxycarbonyl)piperazin-1-yl)-2 1 -Trityl-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclononane (Intermediate 46-9)

[1508]

[1509] This compound was prepared according to the procedure described in Example 44 (Step 10) using Intermediate 46-8 instead of Intermediate 44-9 as the starting material. Yield: 65%.

[1510] ESI-MS: m / z = 805.5, [M+H] + .

[1511] Step 4:1 6 -(4-(tert-Butyloxycarbonyl)piperazin-1-yl)-2 1 -Trityl-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclononane (Intermediate 46-10)

[1512]

[1513] This compound was prepared according to the procedure described in Example 45 (Step 4) using Intermediate 46-9 instead of Intermediate 45-9 as the starting material. Yield: 94%.

[1514] ESI-MS: m / z = 463.2, [M+H] + .

[1515] Step 5: Preparation of compound 46

[1516]

[1517] This compound was prepared according to the procedure described in Example 45 (Step 5) using Intermediate 46-10 instead of Intermediate 45-10 as the starting material. Yield: 61%.

[1518] ESI-MS: m / z = 477.2, [M+H] + .

[1519] 1 H NMR(DMSO-d6,400MHz)δ12.80(s,1H),8.41(s,1H),7.53-7.45(m,2H),7.17(d,J=8.7Hz,1H),7.02-6.95(m,2H),6.64(s ,1H),4.47-4.32(m,2H),4.23-4.17(m,1H),4.10(t,J=11.9Hz,1H),3.32(s,2H),3.09(s,4H),2.61(s,4H),2.37-2.18(m 4H),2.17-2.05(m,1H).

[1520] Example 47

[1521] Compound 47: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 H-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclooctane

[1522]

[1523] Step 1: Preparation of 5-(2-fluoro-6-hydroxyphenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-((tetrahydro-2H-pyran-2-yl)oxy)propoxy)phenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 47-7)

[1524]

[1525] This compound was prepared according to the procedure described in Example 44 (Step 7) using Intermediate A4 instead of Intermediate A2 as the starting material. Yield: 100%.

[1526] ESI-MS: m / z = 863.5, [M+H] + .

[1527] Step 2: Preparation of 5-(2-fluoro-6-hydroxyphenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-hydroxypropyloxy)phenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 47-8)

[1528]

[1529] This compound was prepared according to the procedure described in Example 44 (Step 8) using Intermediate 47-7 instead of Intermediate 44-7 as the starting material. Yield: 91%.

[1530] ESI-MS: m / z = 779.5, [M+H] + .

[1531] Step 3: Preparation of 5-(2-fluoro-6-hydroxyphenyl)-3-(4-((R)-hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-3-(3-bromopropoxy)phenyl)-1-trityl-1H-pyrazolo[4,3-c]pyridazin-6(5H)-one (Intermediate 47-9)

[1532]

[1533] This compound was prepared according to the procedure described in Example 44 (Step 9) using Intermediate 47-8 instead of Intermediate 44-8 as the starting material. Yield: 71%.

[1534] ESI-MS: m / z = 841.3, [M+H] + .

[1535] Step 4: (R)-1 6 -(hexahydropyrazino[2,1-c][1,4]oxazin-8(1H)-yl)-2 1 -Trityl-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclononane (Intermediate 47-10)

[1536]

[1537] This compound was prepared according to the procedure described in Example 44 (Step 10) using Intermediate 47-9 instead of Intermediate 44-9 as the starting material. Yield: 84%.

[1538] ESI-MS: m / z = 761.5, [M+H]+ .

[1539] Step 5: Preparation of compound 47

[1540]

[1541] This compound was prepared according to the procedure described in Example 44 (Step 11) using Intermediate 47-10 instead of Intermediate 44-10 as the starting material. Yield: 64%.

[1542] ESI-MS: m / z = 519.3, [M+H] + .

[1543] 1 H NMR(DMSO-d6,400MHz)δ12.78(s,1H),8.41(s,1H),7.53-7.45(m,2H),7.18(d,J=8.7Hz ,1H),7.02-6.95(m,2H),6.64(s,1H),4.46-4.33(m,2H),4.23-4.17(m,1H),4.15-4.04( m,1H),3.76(d,J=9.6Hz,1H),3.67(d,J=10.5Hz,1H),3.58-3.44(m,2H),3.25(t,J=10.4 Hz, 1H), 3.15 (t, J = 9.3Hz, 1H), 2.80-2.62 (m, 3H), 2.41-2.18 (m, 5H), 2.18-2.06 (m, 1H).

[1544] Example 48

[1545] Compound 48:1 5 -(4-cyanopyridin-3-yl)-2 1 H-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclooctane

[1546]

[1547] Step 1:1 5 -Br-2 1 -(tetrahydro-2H-pyran-2-yl)-3 6 Preparation of 4,8-dioxa-1(1,3)-benzyl-2(3,5)-(6-azaindazole)-3(1,2)-benzylheterocyclooctane (Intermediate 48-1)

[1548]

[1549] This compound was prepared according to the procedures described in Example 18 (steps 1 to 7) using 3-bromo-5-iodophenol instead of 2-bromo-5-iodophenol as the starting material to provide intermediate 48-1.

[1550] ESI-MS: m / z = 524.3, [M+H] + .

[1551] Step 2:1 5 -(4-cyanopyridin-3-yl)-2 1 -(tetrahydro-2H-pyran-2-yl)-3 6 Preparation of 4,8-dioxa-1(1,3)-benzo-2(3,5)-(6-oxo-1H-pyrazolo[4,3-c]pyridazin-5(6H)-yl)-3(1,2)-benzheterocyclooctane (Intermediate 48-2)

[1552]

[1553] This compound was prepared according to the procedure described in Example 8 (Step 5) using Intermediate 48-1 instead of Intermediate 8-4 and (4-cyanopyridin-3-yl)boronic acid pinacol ester instead of Intermediate 8-2 as starting materials to give Intermediate 48-2 in 61% yield.

[1554] ESI-MS: m / z = 548.3, [M+H] + .

[1555] Step 3: Preparation of compound 48

[1556] This compound was prepared according to the procedure described in Example 15 (Step 6) using Intermediate 48-2 instead of Intermediate 15-5 as the starting material to give the title compound (white solid) in 66% yield.

[1557] ESI-MS: m / z = 464.3, [M+H] + .

[1558] 1 H NMR(DMSO-d6,400MHz)δ13.90(s,1H),9.16(s,1H),9.01(s,1H),8.90-8.86(m,2H),8.13(s,1H),8.03(d,J=5.0Hz ,1H),7.80(s,1H),7.46-7.39(m,1H),7.23(s,1H),7.03-6.96(m,2H),4.66-4.58(m,2H),4.18-4.12(m,2H),2.42 -2.32(m,2H).

[1559] The following compounds were synthesized by a method similar to Example 48 using appropriate intermediates:

[1560]

[1561]

[1562] Biological activity detection methods and results:

[1563] 1. HPK1 kinase activity assay

[1564] HPK1 kinase activity is manifested as autophosphorylation activity and phosphorylation of downstream substrates. No additional substrates are required during the autophosphorylation process, and ATP is consumed to produce ADP. The amount of the product is detected using the ADP-Glo ​​reagent and luminescence method to reflect the activity of the kinase.

[1565] Test substance: the compound prepared in the examples of the present invention.

[1566] Prepare compound stock solution: dissolve the test compound in 100% DMSO to make a 10 mM stock solution;

[1567] Prepare 4× kinase reaction buffer:

[1568] name Stock concentration volume Final concentration Tris 1M(25X) 240 μL 40mM <![CDATA[MgCl2]]> 1M(50X) 120 μL 20mM BSA 7.5％(75X) 80 μL 0.1％ DTT 1M(500X) 3μL 0.5mM <![CDATA[ddH2O]]> 5557μL

[1569] Prepare 2× HPK1 kinase solution:

[1570]

[1571] Prepare 4x ATP mix:

[1572]

[1573] Test steps:

[1574] Dilute the test compound stock solution 5-fold with 100% DMSO and perform 4-fold isocratic dilutions in a 96-well dilution plate. Add 1 μL of compound to 49 μL of kinase reaction buffer and shake on a microplate shaker for 20 minutes. Transfer 2 μL of 2× HPK1 kinase solution to a 384-well reaction plate (Greiner, Cat# 784075), add 1 μL of the test compound to the 384-well reaction plate, centrifuge for 1 minute (1000 rpm), and incubate at 25°C for 10 minutes. Transfer 1 μL of 4× ATP mix to the 384-well reaction plate, centrifuge for 1 minute (1000 rpm), and incubate at 25°C for 60 minutes. The final DMSO concentration in all reactions is 0.5%. Transfer 4 μL of ADP-Glo ​​to the 384-well reaction plate, centrifuge for 1 minute (1000 rpm), and incubate at 25°C for 40 minutes. Transfer 8 μL of the detection solution to a 384-well reaction plate, centrifuge for 1 minute (1000 rpm), and incubate for 40 minutes at 25° C. Use a Biotek multi-function plate reader to read the fluorescence signal, and use a four-coefficient nonlinear fitting formula to calculate the IC50 (half-maximal inhibitory concentration) of the compound.

[1575] As shown in the examples, the compounds showed IC50 values ​​in the following range: ++++ = IC50 ≤ 5 nM, +++ = 5 nM <IC50≤50nM,++=50nM<IC50≤500nM,+=IC50> 500nM.

[1576] Table 1 Inhibitory effect of compounds on HPK1 kinase activity

[1577]

[1578]

[1579] The data in Table 1 show that the compounds of the examples of the present invention have a strong inhibitory effect on the activity of HPK1 kinase.

[1580] 2. FLT3 kinase activity test

[1581] Test substance: the compound prepared in the examples of the present invention.

[1582] Prepare compound stocks: Dissolve the test compound in 100% DMSO to make a 10 mM stock solution. Dilute the test compound stock 5-fold with 100% DMSO. Perform 4-fold aliquots in a 96-well dilution plate. Add 1 μL of compound to 39 μL of kinase reaction buffer (1× kinase buffer, 5 mM MgCl2, 1 mM DTT) and shake on a microplate shaker for 20 minutes. Prepare 4× positive controls (4 μL of the first concentration point of the positive drug added to 196 μL of kinase buffer) and 4× negative controls (4 μL of 100% DMSO added to 196 μL of kinase buffer).

[1583] Test steps:

[1584] (1) Prepare 2.5× FLT3 kinase solution (160 pM) with 1× enzyme reaction buffer, transfer 2 μL of kinase solution to a 384-well reaction plate, add 1 μL of the compound to be tested in a gradient concentration to the 384-well reaction plate (Greiner, 784075), centrifuge for 1 minute (1000 rpm), and incubate at 25°C for 10 minutes.

[1585] (2) Prepare 2.5× TK-substrate-biotin (2.5 μM) and 2.5× ATP (2.5 μM) in 1× enzyme reaction buffer, mix well, add 2 μL of TK-substrate-biotin / ATP mixture to the 384 reaction plate, centrifuge for 30 seconds (1000 rpm), and incubate at 25°C for 40 minutes.

[1586] (3) Prepare 2× Sa-XL 665 (125 nM) and 1× TK-antibody-cryptate in HTRF detection buffer. Add 5 μL of the mixture of Sa-XL 665 and TK-antibody-cryptate to each well, centrifuge for 30 seconds (1000 rpm), and incubate at room temperature for 1 hour.

[1587] (4) The fluorescence signal was read using a Biotek multifunctional plate reader, and the IC50 (half maximal inhibitory concentration) of the compound was obtained using a four-coefficient nonlinear fitting formula.

[1588] As shown in the examples, the compounds showed IC50 values ​​in the following range: ++++ = IC50 ≤ 5 nM, +++ = 5 nM <IC50≤50nM,++=50nM<IC50≤500nM,+=IC50> 500nM.

[1589] Table 2 Inhibitory effect of compounds on FLT3 kinase activity

[1590]

[1591]

[1592] The data in Table 2 show that the compounds of the examples of the present invention have a strong inhibitory effect on the activity of FLT3 kinase.

[1593] 3. KDR kinase activity assay

[1594] Test substance: the compound prepared in the examples of the present invention.

[1595] Prepare compound stocks: Dissolve the test compound in 100% DMSO to make a 10 mM stock solution. Dilute the test compound stock 5-fold with 100% DMSO. Perform 4-fold aliquots in a 96-well dilution plate. Add 1 μL of compound to 39 μL of kinase reaction buffer (1× kinase buffer, 5 mM MgCl2, 1 mM DTT) and shake on a microplate shaker for 20 minutes. Prepare 4× positive controls (4 μL of the first concentration point of the positive drug added to 196 μL of kinase buffer) and 4× negative controls (4 μL of 100% DMSO added to 196 μL of kinase buffer).

[1596] Test steps:

[1597] (1) Prepare 2.5× KDR kinase solution (250 pM) in 1× enzyme reaction buffer, transfer 2 μL of kinase solution to a 384-well reaction plate, add 1 μL of the compound to be tested in a gradient concentration to the 384-well reaction plate (Greiner, Cat#784075), centrifuge for 1 minute (1000 rpm), and incubate at 25°C for 10 minutes.

[1598] (2) Prepare 2.5× TK-substrate-biotin (2.5 μM) and 2.5× ATP (12.5 μM) in 1× enzyme reaction buffer, mix well, add 2 μL of TK-substrate-biotin / ATP mixture to a 384-well reaction plate, centrifuge for 30 seconds (1000 rpm), and incubate at 25°C for 40 minutes.

[1599] (3) Prepare 2× Sa-XL 665 (125 nM) and 1× TK-antibody-cryptate in HTRF detection buffer. Add 5 μL of the mixture of Sa-XL 665 and TK-antibody-cryptate to each well, centrifuge for 30 seconds (1000 rpm), and incubate at room temperature for 1 hour.

[1600] (4) The fluorescence signal was read using a Biotek multifunctional plate reader, and the IC50 (half maximal inhibitory concentration) of the compound was obtained using a four-coefficient nonlinear fitting formula.

[1601] As shown in the examples, the compounds showed IC50 values ​​in the following range: ++++ = IC50 ≤ 5 nM, +++ = 5 nM <IC50≤50nM,++=50nM<IC50≤500nM,+=IC50> 500nM.

[1602] Table 3 Inhibitory effect of compounds on KDR kinase activity

[1603]

[1604]

[1605] 4.ELISA detection of IL-2 secretion by Jurkat cells

[1606] Test substance: the compound prepared in the examples of the present invention.

[1607] Test steps:

[1608] Human Jurkat-E6-1 cells were incubated with various concentrations of test compounds in a humidified incubator at 37°C and 5% CO₂ for 30 minutes. The cells were then transferred to a cell culture plate pre-coated with anti-human CD3 antibody. Soluble anti-human CD28 antibody was then added and the cells were stimulated for 24 hours in a humidified incubator at 37°C and 5% CO₂. The cell culture medium was collected by centrifugation and transferred to a 96-well clear microplate (Thermo) pre-coated with anti-human IL-2 antibody. The plates were incubated at room temperature for 2 hours with gentle shaking. The plates were washed four times with wash buffer and the OD values ​​were read using a microplate reader (Molecular Device, i3X) according to the ELISA MAX Deluxe Set Human IL-2 (BioLegend, Cat# 431804) kit protocol. The optimal standard curve was selected using the microplate reader software, and the corresponding concentrations were calculated based on the OD values ​​of the standards. Results are expressed as the percentage (%) of IL-2 secretion by compound-treated versus DMSO-treated cells.

[1609] Table 4 Effects of compounds on IL-2 secretion by human Jurkat cells

[1610]

[1611]

[1612] The data in Table 4 show that compared with the blank control group, which was treated with DMSO, the compounds of the examples of the present invention had a significant promoting effect on the secretion of cytokine IL-2 by Jurkat cells.

[1613] 5.MV-4-11 cell activity test

[1614] The CellTiter-Glo (CTG) luminescence assay was used to detect the effects of the compounds on the activity of MV-4-11 cells.

[1615] Test substance: the compound prepared in the examples of the present invention.

[1616] Test steps:

[1617] In a 96-well flat-bottomed, clear-bottomed cell plate (Corning, Cat#3603), 100 μL of a cell suspension containing 20,000 MV-4-11 cells (human myelomonocytic leukemia cells, obtained from the Chinese Academy of Sciences Cell Bank, Catalog No. SCSP-5031) was added to each well. Then, 100 μL of cell culture medium containing a concentration series of the test compound was added to a total volume of 200 μL. The final compound concentrations were 500 nM, 125 nM, 31.25 nM, 7.813 nM, 1.953 nM, 0.488 nM, 0.122 nM, 0.031 nM, and 0.008 nM, with a final DMSO content of 0.5%. A 100% cell control was established by adding cell culture medium containing 0.5% DMSO and cells but no compound. A culture medium control was established by adding culture medium without cells. The cell plates were incubated in a humidified incubator at 37°C and 5% CO2 for 5 days. After 5 days, the cell plates were removed and centrifuged (1000 rpm) for 5 minutes. 100 μL of supernatant was removed and discarded. 100 μL of CTG reagent (Promega, Cat# G7572) was added to each well and shaken at room temperature for 10 minutes to promote cell lysis. The cells were incubated at room temperature for 30 minutes to allow the luminescent signal to stabilize. Luminescent fluorescence was read using a microplate reader (Molecular Device, i3X). Raw data were analyzed using GraphPad Prism 5.0 software. Dose-effect curves (cell viability %) were obtained by fitting the data using nonlinear S-curve regression, and IC50 values ​​were calculated.

[1618] Table 5 Inhibitory effect of compounds on MV-4-11 cell activity

[1619] Compound IC50(nM) Compound 8 140 Compound 15 2.6 Compound 27 3.3 Compound 31 2.4 Compound 33 60 Compound 39 47

[1620] Unless otherwise defined, the terms used in the present invention have the same meanings as those commonly understood by those skilled in the art.

[1621] The present application has been described above in conjunction with preferred embodiments, but these embodiments are merely exemplary and serve only as an illustration. On this basis, various replacements and improvements can be made to the present application, all of which fall within the scope of protection of the present application. The embodiments described in the present invention are for illustrative purposes only and are not intended to limit the scope of protection of the present invention. Those skilled in the art may make various other replacements, changes and improvements within the scope of the present invention. Therefore, the present invention is not limited to the above-mentioned embodiments, but is only limited by the claims.

Claims

1. A macrocyclic compound of formula (I), or a stereoisomer or a pharmaceutically acceptable salt thereof, in, W is N or CH; X is N, Y is C, Z is CH, and there is a double bond between X and Y; Ring B is a benzene ring; L is independently selected at each occurrence from -O-, -CH2-, -CH(R 1 )-、-C(R 1 )2-、 And one L connected to -O- is not -O-; and any two adjacent L are not -O- at the same time, and are not And not at the same time m mutually connected Ls constitute a linker connecting -O- and the B ring; R 1 Selected from F and C 1-3 alkyl; R 2 Selected from H, F and C 1-3 alkoxy; R 3 Selected from halogen, optionally substituted with 1, 2 or 3 R 31A C 1-3 Alkyl, optionally substituted with 1, 2 or 3 R 31A C 3-6 Cycloalkyl, C 2-6 Alkynyl and cyano groups; R 4 Selected from H, F and C 1-3 alkoxy; R 5 Selected from: 1) H, F, and C 1-3 Alkoxy, 2) optionally substituted with 1 or 2 R 51A phenyl and a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group contains 1 or 2 ring-forming heteroatoms selected from N, O or S, 3) optionally substituted with 1, 2 or 3 R 51B A 5-7 membered monocyclic aliphatic heterocyclic group; R 6 Selected from: 1) H, F, and C 1-3 Alkoxy, 2) One of the following structures, where the chemical bond ends are It indicates that it is connected to other atoms in the structure of formula (I) through this bond: And R 5 and R 6 Not H, F or C at the same time 1-3 alkoxy; The condition is that when (1)n is 0, R 4 and R 5 are all H, and L is independently selected from -O- or -CH2-, or, (2) n is 0, R 4 and R 5 Both are H, and -(L) m -for -(L) m -When connected to the B ring through the bond marked with "*", R 6 Select one of the following structures: R 31A Independently selected from F, hydroxy, C 1-3 Alkoxy, amino, C 1-3 Alkylamino, and cyano; R 51A Independently selected from: 1) Halogen, -OR a1 and cyano, and optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl; R 51B Independently selected from: 1) C 1-3 Alkyl, cyclopropyl, the C 1-3 Alkyl and cyclopropyl are optionally substituted with 1, 2 or 3 R G ;or 2) Two R attached to the same carbon atom 51B Together with the carbon atom, R a1 Each independently selected from: 1) Hydrogen; 2) optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl; R A Selected from H, optionally substituted with 1, 2 or 3 R A11 C 1-3 alkyl, Among them, R A11 is selected from fluoro, hydroxy or cyano; R B Selected from H, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino, optionally substituted with 1, 2 or 3 R B11 C 1-3 Alkyl, cyclopropyl, cyano, Among them, R B11 is selected from fluorine or hydroxyl; wherein, The ring carbon atoms in are optionally substituted with 1, 2 or 3 C 1-3 Alkyl; The ring-forming carbon atoms in are optionally substituted with 1, 2 or 3 R G ; and, two R attached to the same carbon atom B Different from H, hydroxyl, C 1-3 Alkoxy, C 1-3 Alkylamino, R G Selected from oxo, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Fluoroalkoxy, cyclopropyloxy, C 1-3 Alkyl, C 1-3 Fluoroalkyl, cyclopropyl, and cyano groups; E 1 Selected from -CH(R x )-、 where R X Selected from E 2 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(R Y )-、 where R Y Selected from isopropyl, E 3 Selected from -C(CH3)2-, -CH(CH2CN)-, E 4 Selected from carbonyl, -CH2-, -C(CH3)2-, -CH(CH2CN)-, E 5 Selected from -CH2-, -C(CH3)2-, -CF2-, where R Z Selected from H and C 1-3 alkyl; The E ring is a 3-membered saturated alicyclic ring or a 4-6-membered saturated alicyclic heterocyclic ring, wherein the saturated alicyclic heterocyclic ring contains 1 or 2 ring-forming heteroatoms selected from N and O, and the ring-forming carbon atoms in the saturated alicyclic heterocyclic ring are optionally substituted with 1, 2 or 3 C 1-3 Alkyl, saturated aliphatic heterocyclic ring, if present, is optionally substituted with a ring nitrogen atom of R A ; m is 4, 5 or 6; n is 0, 1, or 2; q is 1 or 2.

2. The macrocyclic compound according to claim 1, wherein W is CH, R 2 Selected from H and F.

3. The macrocyclic compound according to claim 2, wherein The compound has the structure of formula (IIa), Among them, -(L) m -Selected from one of the following structures: R 2 Selected from H and F; R 3 Located in R 2 Ortho or meta, where (1) when R 3 Located in R 2 When the adjacent position of R 3 is selected from F, Cl, methyl or cyclopropyl; (2) when R 3 Located in R 2 When the meta position is 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are optionally substituted with one selected from hydroxy, methoxy, methylamino, dimethylamino or Substituents; R 4 Selected from H, F and C 1-3 Alkoxy, R 5 Selected from H, F and C 1-3 Alkoxy, and R 4 and R 5 There is only one selected from F and C 1-3 Alkoxy, or R 4 and R 5 All are H; R 6 Select one of the following structures: in, R A Selected from C 1-3 Alkyl, 1, 2 or 3 fluorine-substituted C 1-3 Alkyl and 1, 2 or 3 hydroxy substituted C 1-3 alkyl; R B selected from H, hydroxyl, C substituted with 1, 2 or 3 hydroxyl groups 1-3 alkyl, described The ring carbon atoms in are optionally substituted with 1, 2 or 3 hydroxyl or methyl groups; and the two R B Different from H, hydroxyl, E 4 is selected from carbonyl and -CH2-; The E ring is selected from one of the following structures: wherein the ring-forming carbon atoms are unsubstituted; q is selected from 1 and 2.

4. The macrocyclic compound according to claim 3, wherein R 6 Select one of the following structures:

5. The macrocyclic compound according to claim 2, wherein The compound has the structure of formula (IIa), Among them, -(L) m -Selected from one of the following structures: R 2 Selected from H and F; R 3 Located in R 2 Ortho or meta, where (1) when R 3 Located in R 2 When the adjacent position of R 3 is selected from F, Cl, methyl or cyclopropyl, (2) when R 3 Located in R 2 When the meta position is 3 is selected from methyl, ethyl, cyclopropyl, ethynyl and cyano, wherein the methyl and ethyl groups are optionally substituted with one selected from hydroxy, methoxy, methylamino, dimethylamino or Substituents; R 4 Selected from H, F and C 1-3 Alkoxy, R 6 Selected from H, F and C 1-3 Alkoxy, and R 4 and R 6 There is only one selected from F, or R 4 and R 6 All are H; R 5 Selected from: 1) optionally substituted with 1 or 2 R 51A pyridyl, imidazolyl and pyrazolyl, 2) optionally substituted with 1, 2 or 3 R 51B A 5-6 membered monocyclic aliphatic heterocyclic group; Among them, R 51A Selected from halogen, -OR a1 、C 1-3 Alkyl, 1, 2 or 3 fluorine-substituted C 1-3 alkyl, cyclopropyl, and cyano groups; R 51B Selected from: 1) methyl, difluoromethyl, trifluoromethyl, isopropyl, cyclopropyl, or 2) Two R attached to the same carbon atom 51B Together with the carbon atom, Among them, R a1 Each independently selected from: 1) Hydrogen; 2) optionally substituted with 1, 2 or 3 R G C 1-3 Alkyl and cyclopropyl; R G Selected from oxo, F, hydroxyl, C 1-3 Alkoxy, C 1-3 Fluoroalkoxy, cyclopropyloxy, C 1-3 Alkyl, C 1-3 Fluoroalkyl, cyclopropyl and cyano.

6. The macrocyclic compound according to claim 5, wherein The 5-6 membered monocyclic aliphatic heterocyclic group is selected from:

7. The macrocyclic compound according to claim 5, wherein R 5 Select one of the following structures:

8. The macrocyclic compound according to claim 5, wherein R 5 Select one of the following structures: Optionally substituted with 1 R 51A The pyridyl group, R 51A selected from cyano; Replace with 1 R 51B A 5-6 membered monocyclic aliphatic heterocyclic group selected from the following: Among them, R 51B It is a methyl group.

9. The macrocyclic compound according to claim 1, wherein The compound has the structure of formula (IIIa), Among them, -(L) m -Selected from one of the following structures: in, 1) When R 4 and R 5 There is only one selected from F and C 1-3 Alkoxy, the other is H, R 6 Select one of the following structures: 2) When R 4 and R 6 When both are H, R 5 Selected from: 1) optionally substituted with 1 or 2 R 51A substituted pyridyl, imidazolyl and pyrazolyl groups, 2) optionally substituted with 1, 2 or 3 R 51B The following ring systems: in, R 51A independently selected from F, Cl, methoxy, methyl, cyclopropyl and cyano; R 51B Selected from: 1) methyl and isopropyl, or 2) Two R attached to the same carbon atom 51B Together with the carbon atom, 10. The macrocyclic compound according to claim 1, wherein The compound has the structure of formula (IIIa), Among them, -(L) m -Selected from one of the following structures: in, R 4 Selected from H, F and C 1-3 Alkoxy, R 5 Selected from H, F and C 1-3 Alkoxy, and R 4 and R 5 There is only one selected from F and C 1-3 Alkoxy, or R 4 and R 5 All are H, R 6 Select one of the following structures:

11. The macrocyclic compound according to claim 1, wherein The compound has the structure of formula (IIIa), Among them, -(L) m -Selected from one of the following structures: R 4 and R 5 All are H, R 6 Select one of the following structures: in, R B selected from H, hydroxyl, C substituted with 1, 2 or 3 hydroxyl groups 1-3 alkyl, described The ring carbon atoms in are optionally substituted with 1, 2 or 3 hydroxyl or methyl groups; and the two R B Different from H, hydroxyl, The E ring is selected from one of the following structures: wherein the ring-forming carbon atoms are unsubstituted; q is selected from 1 and 2.

12. The macrocyclic compound according to claim 1, wherein The compound has the structure of formula (IIIa), Among them, -(L) m -Selected from one of the following structures: R 4 and R 5 All are H, R 6 Select one of the following structures:

13. The compound according to claim 1, wherein -(L) m - one selected from the following structures, 14. The compound according to claim 1, wherein R 2 is selected from F and methoxy; Or, R 2 Selected from F; Or, R 2 Selected from H.

15. The compound according to claim 1, wherein There is only one R 3 , and is in R 2 Ortho position, R 3 is selected from F, Cl, methyl or cyclopropyl; Or, there is only one R 3 , and is in R 2 The meta position, R 3 is selected from F, Cl, methyl, difluoromethyl, trifluoromethyl, ethyl, isopropyl, cyclopropyl, ethynyl and cyano, wherein the methyl, ethyl, isopropyl and cyclopropyl are unsubstituted or substituted with one selected from hydroxy, C 1-3 Alkoxy, amino, C 1-3 Alkylamino, or cyano substituents; the ethynyl group is unsubstituted or substituted with one selected from methyl, cyclopropyl, Substituents; Alternatively, n is 0.

16. The compound according to claim 1, wherein There is only one R 3 , and is in R 2 Ortho position, R 3 Selected from F and Cl.

17. The compound according to claim 1, wherein R 4 Selected from H and F; Or, R 4 selected from methoxy; Or, R 4 Selected from F; Or, R 4 Selected from H; Or, R 5 Selected from H and F; Or, R 5 Selected from F; Or, R 5 Selected from H.

18. The compound according to any one of claims 1 to 12, wherein The compounds described do not include the following compounds:

19. A macrocyclic compound selected from the group consisting of the following compounds, or a stereoisomer or a pharmaceutically acceptable salt thereof:

20. A pharmaceutical composition comprising the compound according to any one of claims 1 to 19 or a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier.

21. Use of the compound according to any one of claims 1 to 19 or a pharmaceutically acceptable salt or stereoisomer thereof, or the pharmaceutical composition according to claim 20, in the preparation of a medicament for preventing or treating a disease mediated by protein kinase, wherein the disease is one or more of a tumor, myelodysplastic syndrome, and a disease induced by a virus; wherein, The protein kinase is selected from one or more of HPK1, FLT3 and KDR; wherein the tumor is one or more of chronic or acute leukemia, lymphoma, multiple myeloma, myeloproliferative neoplasms, lung cancer, hepatocellular carcinoma, gallbladder cancer, bile duct cancer, gastric cancer, colorectal cancer, small intestinal leiomyosarcoma, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, fallopian tube cancer, vaginal cancer, vulvar cancer, malignant teratoma, pancreatic cancer, neuroendocrine tumor, nasopharyngeal cancer, oral cancer, laryngeal cancer, hypopharyngeal cancer, esophageal cancer, thyroid cancer, pheochromocytoma, endocrine pancreatic tumor, kidney cancer, bladder cancer, malignant brain tumor, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, fibrosarcoma of bone, Ewing's sarcoma, myxoma, malignant thymoma, malignant peripheral nerve sheath tumor, prostate cancer, testicular cancer, penile cancer, urethral cancer and benign and malignant skin tumors; The virus is one or more of hepatitis virus, human immunodeficiency virus, human papillomavirus, herpes simplex virus, measles virus, norovirus, bocavirus, coxsackievirus, Ebola virus, enterovirus, lymphocytic meningitis virus, influenza virus, SARS virus and new coronavirus.

22. The use according to claim 21, wherein the tumor is primary CNS lymphoma, intrahepatic cholangiocarcinoma, or esophageal cardia cancer.

23. The use according to claim 21, wherein the skin malignancy is one or more of squamous cell carcinoma, basal cell carcinoma, and malignant melanoma.

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