Macrocyclic compound as kinase inhibitor, and use thereof

Abstract

The present disclosure relates to a macrocyclic compound represented by formula I, a pharmaceutical composition containing the compound or a pharmaceutically acceptable salt, prodrug, solvate, polymorph, isomer and stable isotopic derivative thereof, a preparation method therefor, and a use thereof as a TRK, ROS1, c-Met, CSF1 and / or ALK inhibitor.

Description

Macrocyclic compounds as kinase inhibitors and their uses Technical Field

[0001] This disclosure relates to macrocyclic compounds of Formula I, pharmaceutically acceptable salts thereof, prodrugs, solvates, polymorphs, isomers, or stable isotopic derivatives thereof, and pharmaceutical compositions comprising them, methods of their preparation, and their use as inhibitors of TRK, ROS1, c-Met, CSF1, and ALK. This disclosure also relates to the use of them to treat TRK, ROS1, c-Met, CSF1, and ALK-mediated diseases, such as cancer and ubiquitination, and methods of using them to treat such diseases. Background Technology

[0002] Kinases play an important role in regulating cell signaling pathways by catalyzing protein phosphorylation, and are essential for cell growth, proliferation and survival [a): D. Bossemeyer. Protein kinases - structure and function. 1995, FEBS Let., 369, 57-61. b). Stevan R. Hubbard et al. Protein Tyrosine Kinase Structure and Function. Ann. Rev. of Biochem., 2000, 69, 373-398. The cumulative genetic and epigenetic modifications of kinases (such as site mutations, deletions, insertions, amplifications, and translocations) lead to abnormal activation of cell signaling pathways, thereby driving cancer cell growth, proliferation, metastasis, and survival [a]: Torkamanani A. et al. Cancer driver mutations in protein kinase genes. Can. Lett., 2009, 281(2), 117-127. [b]: Lahiry P. et al. Kinase mutations in human disease: interpreting genotype-phenotype relationships. Nat. Rev. Gen., 2010, 11, 60-74. Targeting different kinases has proven to be a very successful approach for treating various diseases, including cancer and autoimmune diseases. [Richard A. Ward, Frederick W. Goldberg (Edited). Kinase Drug Discovery (Drug Discovery, Vol. 19), 2011, RSC publishing.] Inhibition of TRKS, ROS1, and ALk has proven to be an effective treatment option for various cancers marked by their respective gene fusions. FDA-approved larotrectinib and Entrectinib have both demonstrated high efficacy in clinical practice. However, their effectiveness is increasingly challenged by cumulative resistance, marked by TRK mutations (Cancer Discov; 2018, 8(10); 1227–36). To overcome the resistance problem, second-generation therapies, represented by TPX-0005 (CN 106170289B) and LOXO-195 (CN 108697708 A), are under active development.

[0003] TRK: Tropomyosin receptor kinase (TRK) is encoded by the NTRK gene and plays a crucial role in the development and normal function of the nervous system. The TRK family consists of TRKA, TRKB, and TRKC. They are high-affinity receptors for neurotrophic factors (NTs) and are activated by their preferred neurotrophic factors (NGF to TrkA, brain-derived neurotrophic factor [BDNF] and NT4 / 5 to TrkB, and NT3 to TrkC) [a]: R. Klein, et al. The trk protooncogene encodes a receptor for nerve growth factor. Cell, 1991, 189-197. [b]: BLHempstead et al. High-affinity NGF binding requires coexpression of the trk protooncogene and the low-affinity NGF receptor. Nature 1991, 350, 678-683. In fact, TRK was initially cloned as an oncogene fused with the tropomyosin gene in its extracellular domain [M. Barbacid et al. The trk family of tyrosine protein kinase receptors, Biochim. Biophys. Acta, 1991, 1072, 115-127.]. Since the first discovery of fusion genes in colorectal cancer in 1986, more than 80 fusion genes have now been identified in adult and pediatric tumors [A. Amatu et al. Tropomyosin receptor kinase (TRK) biology and the role of NTRK gene fusions in cancer. Ann. of Oncol., 2019, 30(Suppl8): viii5-viii15.Some of these gene aberrations lead to the expression of constitutively active fusion proteins, which become carcinogenic drivers and have become unique targets for the treatment of related cancers such as non-small cell lung cancer, papillary carcinoma, medullary thyroid carcinoma, breast cancer and colorectal cancer [Carol J. Thiele, et al. “On Trk”-the TrkB signal transduction pathway is an increasingly important target in cancer biology. Clin Cancer Res., 2009, 15(19), 5962–5967.].

[0004] ROS1

[0005] The aberrant ROS1 protein encoded by the fusion proto-oncogene ROS1 with an unknown ligand belongs to the insulin receptor family [H. Matsushime, et al. Human c-ros-1 gene ho mologous to the v-ros sequence of UR2 sarcoma virus encodes for a transmembrane receptor-like molecule. Mol. Cell Biol. 1986, 6(8), 3000-3004]. The fusion protein ROS1 can autophosphorylate and then mediate tumor progression through the mitogen-activated protein kinase (MAPK) pathway or RAS phosphorylation [Sato H, Schoenfeld AJ, Siau E, et al. MAPK pathway alterations correlate with poor survival and drive resistance to therapy in patients with lung cancers driven by ROS1 fusions. Clin Cancer Res. 2020, 26(12), 2932-2945.]. ROS1 fusions have been reported in 2.54% of lung adenocarcinomas, with even higher frequencies in spitzoid tumors and inflammatory myofibroblastic tumors [Alexander Drilon, et al. ROS1-dependent cancers-biology, diagnostics and therapeutics. Nat. Rev. Clin. Oncol. 2021, 18(1), 35-55.]. To date, approximately 26 genes have been found to fuse with ROS1, such as RET and ALK. Most fusion genes retain the dimerization domain in the fusion protein, which may lead to constitutive ROS1 tyrosine kinase activation. Numerous studies have shown that constitutively active fusions of ROS1 protein are present in a variety of human cancers, such as glioblastoma, non-small cell lung cancer (NSCLC), cholangiocarcinoma, gastric adenocarcinoma, colorectal cancer, ovarian cancer, inflammatory myofibroblastoma, angiosarcoma, and epithelioid angioendothelioma [Arnaud Uguen, Marc De Braekeleer. ROS1 fusions in cancer: a review. Future Oncol. 2016, 12(16), 1911-28.].Currently, ROS1 fusion gene therapy has made significant progress in clinical practice, with several drugs targeting ROS1 fusion genes approved, including first-generation small molecules crizotinib and ceritinib, and second-generation macrocyclic molecules repotrectinib and talectrectinib, for the treatment of ROS1-positive NSCLC [a]. Shizhe Li et al. Current treatment and novel insights regarding ROS1-targeted therapy in malignant tumors. Cancer Med. 2024, 13(8), e7201. b). Jessica J. Lin, Alice T. Shaw. Recent Advances in Targeting ROS1 in Lung Cancer. J. of Thoracic Oncol., 2017, 12(11), 1611-1625. However, like other kinase inhibitors, new challenges such as drug resistance and adverse reactions (including central nervous system metastasis) continue to test human wisdom, and the search for new therapies with better properties remains a long and arduous task.

[0006] ALK:

[0007] Anaplastic lymphoma kinase (ALK), also known as ALK tyrosine kinase receptor or CD246, is a neurotransmitter. In humans, normal ALK expression is relatively limited in the fetal central and peripheral nervous systems, which is crucial for fetal nervous system development and function [a) Iwahara T, Wen D, et al. molecular characterization of ALK, a receptor tyrosine kinase expressed specifically in the nervous system. Oncogene. 1997, 14, 439-449. b) Chiarle R et al. The anaplastic lymphoma kinase in the pathogenesis of cancer. Nat. Rev. Cancer. 2008; 8:11e23.]. It is classified into the insulin receptor (IR) superfamily because the tyrosine kinase domain of human ALK is highly similar to that of the insulin receptor, forming a subgroup of the RTK superfamily along with leukocyte tyrosine kinase (LTK). ALK fusion was first discovered in anaplastic large cell lymphoma (ALCL) cells [Morris SW, Kirstein MN, et al. Fusion of a kinase gene, ALK, to a nucleolar protein gene, NPM, in non-Hodgkin's lymphoma. Science. 1994, 263(5151): 1281-1284.], and was caused by the translocation fusion gene (2;5)(p23:q35), which encodes the fusion protein NPM-ALK, in which the kinase domain of ALK is fused to the N-terminus of nucleolar phosphatase (NPM) [Shiota M et al. Hyperphosphorylation of a novel 80kDa protein-tyrosine kinase similar to Ltk in a human Ki-1 lymphoma cell line, AMS3. Oncogene. 1994, 9(6): 1567-1574.]. ALK consists of an extracellular ligand-binding domain, a transmembrane domain, and an intracellular tyrosine kinase domain.NPM dimerization constitutively activates the ALK kinase domain [a] Morris SW, Naeve C, et al. ALK, the chromosome 2 gene locus altered by the t(2;5) in non-Hodgkin's lymphoma, encodes a novel neural receptor tyrosine kinase that is highly related to leukocyte tyrosine kinase (LTK). Oncogene. 1997, 14(18), 2175-2188. [b] Iwahara T, Fujimoto J, et al. molecular characterization of ALK, a receptor tyrosine kinase expressed specifically in the nervous system. Oncogene. 1997, 14, 439-449. More than twenty different ALK translocation partners have been identified in various cancers, including ALCL, inflammatory myofibroblastic tumor (IMT), non-small cell lung cancer (NSCLC), colorectal cancer (CRC), breast cancer, and other cancers [a) Jessica J. Lin et al. Targeting ALK: Precision Medicine Takes On Drug Resistance. Cancer Discov. 2017, 7(2), 137-155. b) Xue Du et al. ALK-rearrangement in non-small-cell lung cancer (NSCLC). Thorac Cancer. 2018, 9(4), 423-430.]. ALK has become an attractive and effective molecular target for cancer treatment development due to its important role in hematopoietic tumors, solid tumors, and mesenchymal tumors.

[0008] As experience accumulates, tumor cells will inevitably develop drug resistance and drive metastasis or recurrence by generating new mutations and gene amplification. Therefore, potent inhibitors are still needed to combat these tumors [Richard A Ward, Frederick W Goldberg (Edited). Kinase Drug Discovery (Drug Discovery, Vol. 19), 2011, RSC publishing]. The compounds described herein address the resistance problem by inhibiting multiple kinases in the structure-associated oncogenic proteome [Fleur Broekman et al. Tyrosine kinase inhibitors: Multi-targeted or single-targeted? World J Clin Oncol. 2011, 2(2), 80-93.]. This disclosure will detail a unique class of macrocyclic compounds that target the aforementioned kinases. Summary of the Invention

[0009] This disclosure provides compounds, isomers thereof, prodrugs, solvates, isotopic derivatives or pharmaceutically acceptable salts as shown in Formula I, which can be used as inhibitors of TRK, ROS1, cMet and / or ALK kinases.

[0010] in,

[0011] R1 is selected from H, R'-S-, R'-O-, and C. 1-6 C substituted with alkyl and halogen or cyano groups 1-6 Alkyl; R' is selected from C 1-6 alkyl;

[0012] R2 is selected from H and C. 1-6 C substituted with alkyl, halogen or cyano groups 1-6 Alkyl, amino, -NHC 1-6 Alkyl, -N(C) 1-6 alkyl)2-, -SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl groups, -CONH2, -CONR'R” and C 1-6 Alkoxycarbonyl; wherein "NR'R" is selected from C 1-6 alkyl.

[0013] R3 and R4 are independently selected from H and C. 1-6 alkyl;

[0014] R5 and R6 are independently selected from H and C. 1-6Alkyl groups, or R5 and R6, can be linked to form a five- or six-membered heterocycle, which may optionally be substituted with a halogen or have an oxygen heterocycle atom;

[0015] R7 can be one to four independently selected from H, halogen, -CN, C 1-6 C substituted with alkyl, halogen or cyano groups 1-6 Alkyl substituents, C 1-6 Alkoxy;

[0016] X is selected from -CH- and N;

[0017] L is -CHR 10 -CHR 11 -or-CHR 10 -CH2CHR 11 -, where R 10 and R 11 Independently selected from H and C 1-6 alkyl.

[0018] According to some embodiments, this disclosure provides compounds, isomers thereof, prodrugs, solvates, isotope derivatives, or pharmaceutically acceptable salts as shown in Formula I.

[0019] in,

[0020] R1 is selected from H, R'S-, C 1-6 Alkyl and F-substituted C 1-6 Alkyl; R' is selected from C 1-6 alkyl;

[0021] R2 is selected from H and C. 1-6 Alkyl, F-substituted C 1-6 Alkyl, amino and C 1-6 Alkoxycarbonyl;

[0022] R3 and R4 are independently selected from H and C. 1-6 alkyl;

[0023] R5 and R6 are independently selected from H and C. 1-6 Alkyl groups, or R5 and R6, can be linked to form a five- or six-membered saturated heterocycle, which may optionally be substituted with F or have -O- heterocyclic atoms;

[0024] R7 can be one to four independently selected from H, halogen, -CN, C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl substituents (i.e., R7 can substitute 1 to 4 positions on the aromatic ring or 1 to 3 positions on the heteroaromatic ring);

[0025] X is selected from -CH- and N;

[0026] L is -CHR 10 -CHR 11 -or-CHR 10 -CH2CHR 11 -, where R 10 and R 11 Independently selected from H and C 1-6 alkyl;

[0027] In some implementations, R1 is H.

[0028] In some implementations, R1 is selected from H, R2 is selected from H, and C. 1-6 Alkyl, amino and C 1-6 Alkoxycarbonyl group.

[0029] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups.

[0030] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, with both R3 and R4 being H.

[0031] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 is H and R6 is C. 1-6 alkyl.

[0032] In some embodiments, R1 is selected from H, R2 is selected from H, C1-6 alkyl and amino, R3 and R4 are both H, and R5 is C 1-6 Alkyl group and R6 is H.

[0033] In some embodiments, R1 is selected from H, R2 is selected from H, C1-6 alkyl and amino, R3 and R4 are both H, and R5 and R6 are linked to form a bond to form a five- or six-membered saturated heterocycle, which may optionally be substituted with F, or the heterocycle may have -O- heterocycle atoms.

[0034] In some embodiments, R1 is selected from H, R2 is selected from H, C1-6 alkyl and amino, R3 and R4 are both H, or R5 and R6 are connected to form a five-membered saturated nitrogen-containing heterocycle.

[0035] In some embodiments, R1 is selected from H, R2 is selected from H, C1-6 alkyl and amino, R3 and R4 are both H, R5 is H and R6 is C 1-6 Alkyl group, R7 is selected from H, F, CN and Br, and can substitute for more than one position on the aromatic or heteroaromatic ring, such as 1, 2, 3 or 4 positions. For example, R7 is at the para or meta position of the oxygen atom.

[0036] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, and R5 is C. 1-6 Alkyl group, R6 is H, R7 is independently selected from H, halogen, -CN group, and can replace one or more positions of aryl or heteroaryl ring.

[0037] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 and R6 are linked to form a bond to form a five-membered saturated heterocycle, and R7 is independently selected from H, halogen, -CN group, and can replace more than one position of aryl or heteroaryl ring.

[0038] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 is H and R6 is C. 1-6 Alkyl group, R7 is selected from H, F, CN and Br, and can substitute for more than one position of the aromatic or heteroaromatic ring, L is -CHR. 10 -CHR 11 -, where R 10 It is -CH3 and R 11 For H or R 10 For H and R 11 It is CH3.

[0039] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, and R5 is C. 1-6 The alkyl group has R6 as H, R7 is independently selected from H, halogen, and -CN group, and can substitute for more than one position of the aryl or heteroaryl ring, and L is -CHR. 10 -CHR 11 -, where R 10 It is -CH3 and R 11 For H or R 10 For H and R 11 It is CH3.

[0040] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 and R6 are linked to form a five-membered saturated heterocycle, R7 is independently selected from F and -CN groups, which can substitute for more than one position of the aryl or heteroaryl ring, and L is -CHR. 10 -CHR 11 -where R 10 It is -CH3 and R 11 For H or R 10 For H and R 11 It is CH3.

[0041] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 is H and R6 is C. 1-6 Alkyl group, R7 is selected from F and CN, and can replace one or more positions on the aromatic or heteroaromatic ring, L is -CHCH3-CH2, and X is a carbon atom.

[0042] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 and R6 are linked to form a five-membered saturated heterocycle, R7 is independently selected from F and -CN groups, which can replace one or more positions of the aryl or heteroaryl ring, L is -CHCH3-CH2-, and X is N.

[0043] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 is H, and R6 is C. 1-6 Alkyl group, R7 is F, L is -CHCH3-CH2, and X is a carbon atom.

[0044] In some implementations, R1 is selected from H, and R2 is selected from H and C. 1-6 Alkyl and amino groups, R3 and R4 are both H, R5 and R6 are linked to form a five-membered saturated heterocycle, R7 is F, L is -CHCH3-CH2-, and X is N.

[0045] In some embodiments, this disclosure provides the following compounds represented by Formula I, pharmaceutically acceptable salts thereof, prodrugs, solvates, polymorphs, isomers, and stable isotope derivatives:

[0046] R1 is selected from H;

[0047] R2 is selected from H, -NH2, C 1-6 Alkyl and C 1-6 Alkoxycarbonyl;

[0048] R3 and R4 are independently selected from H;

[0049] R5 and R6 are independently selected from H and C. 1-6 Alkyl groups, or R5 and R6, can be linked together to form five- or six-membered saturated nitrogen-containing heterocycles;

[0050] R7 consists of 1-4 independent selections from H, halogens, and -CN;

[0051] X is selected from -CH- and N;

[0052] L is -CHR 10 -CHR 11 -、-CHR10 -CH2CHR 11 -, where R 10 and R 11 Independently selected from H and C 1-6 alkyl.

[0053] In another embodiment, this disclosure relates to the use of any of the following compounds, pharmaceutically acceptable salts, prodrugs, solvates, polymorphs, isomers, and stable isotope derivatives:

[0054] (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0055] (3S,6S,Z)-4 5 -Fluoro-12,3,6-trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0056] (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine

[0057] (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 methyl carboxylate

[0058] (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0059] (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6-formonitrile

[0060] (S,Z)-4 6 -bromo-2-ethyl-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0061] (S,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0062] (S,Z)-2-Ethyl-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0063] (2 2 R,5S,Z)-3 5 -Fluoro-5-methyl-1 3 H-4-oxa-7,9-diaza-1(5,3)-imidazo[4,5-b]pyridine-3(3,2)-pyridine-2(1,2)-pyrrolidinecyclonon-8-ene

[0064] (S,Z)-2-ethyl-4 5 -Fluoro-12,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0065] (R,Z)-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0066] (S)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,9,11-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocycloundecane-10-ene

[0067] (R,Z)-4 6-bromo-2-ethyl-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0068] (S,Z)-4 6 -bromo-2-ethyl-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0069] (R,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0070] (S,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0071] (R,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0072] (R,Z)-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0073] (S,Z)-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0074] (S,Z)-2-Ethyl-7-methyl-1 3H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0075] (R,Z)-2-ethyl-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0076] (2 2 R,6R,Z)-3 5 -Fluoro-6-methyl-1 3 H-4-oxa-7,9-diaza-1(5,3)-imidazo[4,5-b]pyridine-3(3,2)-pyridine-2(1,2)-pyrrolidinecyclonon-8-ene

[0077] (2 2 R,6S,Z)-3 5 -Fluoro-6-methyl-1 3 H-4-oxa-7,9-diaza-1(5,3)-imidazo[4,5-b]pyridine-3(3,2)-pyridine-2(1,2)-pyrrolidinecyclonon-8-ene

[0078] (R,Z)-2-ethyl-4 5 -Fluorine-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0079] (S,Z)-2-ethyl-4 5 -Fluorine-1 2 ,6-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0080] (S,Z)-2-ethyl-4 5 -Fluorine-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0081] (S,Z)-4 6 -bromo-2-ethyl-1 2 ,6-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0082] (R,Z)-4 6 -bromo-2-ethyl-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0083] (S,Z)-4 6 -bromo-2-ethyl-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0084] (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine

[0085] (S,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine

[0086] (3S,6S,E)-4 5 -Fluoro-3,6-dimethyl-9-methylthio-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0087] (3R,6S,Z)-44-fluoro-3,6-dimethyl-13H-5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]piperidine-4(1,2)benzenecyclodecanecyclo-9-ene

[0088] (3R,6S,Z)-3,6-dimethyl-13H-5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 5 -formonitrile

[0089] (3R,6S,Z)-3,6-dimethyl-45-(trifluoromethyl)-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzonocyclodecane-9-ene

[0090] (3R,6S,Z)-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2-benzocyclodecane-9-ene

[0091] (R,Z)-45-fluoro-3-methyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2-benzocyclodecane-9-ene

[0092] (3R,6S,Z)-45-chloro-3,6-dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzocyclodecane-9-ene

[0093] (3R, 6S, Z)-43,4-5-difluoro-3,6-dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2-benzocyclodecane-9-ene

[0094] ((3R, 6S, Z)-45,3,6-trimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2)-benzocyclodecane-9-ene

[0095] (3R, 6S, Z)-4 5 -Methoxy-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene.

[0096] (3S,6S,Z)-4 5 -Fluoro-3,6,9-trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzenecyclodecane-9-ene

[0097] (3S,6S,Z)-4 5 -Fluoro-9-isopropyl-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene

[0098] (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-9-propyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene

[0099] (3S,6S,Z)-4 5 -Fluoro-9-isopropyl-1 2 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzylcyclodecane-9-ene

[0100] (3S,6S,Z)-4 5 -Fluorine-1 2 3,6,9-Tetramethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-pyrido[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene

[0101] (3S,6S,Z)-4 5 -Fluorine-1 2 3,6-Trimethyl-9-propyl-1 3 H-5-O-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene

[0102] (3S,6S,Z)-4 5 -Fluorine-1 7 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzenecyclodecane-9-ene

[0103] (3S,6S,Z)-4 5 -Fluorine-1 7 3,6,9-Tetramethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene

[0104] (3S,6S)-4 5 -Fluorine-1 6 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene

[0105] According to another aspect of this application, this application provides a pharmaceutical composition comprising a compound of Formula I as described in this application, an isomer thereof, a prodrug, a solvate, an isotope derivative or a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

[0106] According to another aspect of this application, this application also relates to the use of compounds of Formula I, isomers thereof, prodrugs, solvates, isotope derivatives or pharmaceutically acceptable salts thereof, in the preparation of medicaments that inhibit TRK, ROS1, c-Met, CSF1 and / or ALK.

[0107] According to another aspect of this application, this application also relates to the use of compounds of Formula I, isomers thereof, prodrugs, solvates, isotope derivatives or pharmaceutically acceptable salts, in the preparation of medicaments for the prevention and treatment of diseases mediated by TRK, ROS1, c-Met, CSF1 and / or ALK (such as pain, cancer and ubiquitination).

[0108] According to another aspect of this application, this application provides a compound, its pharmaceutically acceptable salt, solvate, polymorph, or prodrug, for inhibiting TRK, ROS1, c-Met, CSF1, and / or ALK. It can be used to treat diseases mediated by TRK, ROS1, c-Met, CSF1, and / or ALK (such as pain, cancer, and ubiquitination).

[0109] According to another aspect of this application, this application provides a method for regulating protein kinase activity, comprising contacting the protein kinase with the aforementioned compound or a pharmaceutically acceptable salt, solvate, polymorph, or prodrug thereof. This method can be used in vivo or in vitro. Preferably, the protein kinase is selected from TRK, ROS1, c-Met, CSF1, and / or ALK.

[0110] According to another aspect of this application, this application provides a method for treating diseases related to protein kinase activity, the method comprising administering the above-mentioned compound or a pharmaceutically acceptable salt, solvate, polymorph, or prodrug thereof to an individual in need.

[0111] According to another aspect of this application, this application provides a method for treating diseases related to protein kinase activity, the method comprising administering the above-mentioned compound or a pharmaceutically acceptable salt, solvate, polymorph or prodrug thereof in combination with at least one therapeutic agent to an individual in need.

[0112] According to some embodiments of this application, the diseases related to protein kinase activity described in this application include tumors.

[0113] Invention Details

[0114] The chapter headings used in this document are for organizational purposes only and should not be construed as limiting the subject matter. All references or portions thereof cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and papers, are incorporated herein by reference in their entirety.

[0115] Some chemical terms

[0116] Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent applications, and publications cited in this document are incorporated herein in their entirety through reference.

[0117] It should be understood that the above summary and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of this application. In this application, unless otherwise specifically stated, the singular is used to include the plural. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the term “comprising” and other forms such as “including,” “containing,” and “containing” are non-limiting descriptions.

[0118] Definitions of standard chemical terms can be found in the references (including Carey and Sundberg, "Advanced Organic Chemistry 4th Edition," Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise stated, conventional methods within the scope of the art, such as mass spectrometry, NMR, IR, UV / Vis spectroscopy, and pharmacological methods, are used. Unless specifically defined, the terminology used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and pharmaceutical and medicinal chemistry is known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and in the treatment of patients. For example, reactions and purifications can be carried out using the manufacturer's instructions for use of kits, or in accordance with methods known in the art or the descriptions in this application. The techniques and methods described herein are generally carried out according to conventional methods well known in the art, based on the descriptions in the various summary and more specific references cited and discussed in this specification. In this specification, groups and their substituents can be selected by those skilled in the art to provide stable structural moieties and compounds.

[0119] When a substituent is described using a conventional chemical formula written from left to right, it also includes chemically equivalent substituents obtained when the structural formula is written from right to left. For example, CH2O is equivalent to OCH2.

[0120] The terms “optional / arbitrary” or “optionally / arbitrarily” mean that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation. For example, “optionally substituted alkyl” means “unsubstituted alkyl” (an alkyl group not substituted by a substituent) or “substituted alkyl” (an alkyl group substituted by a substituent).

[0121] The C1-C used in this article n Including C1-C2, C1-C3, ..., C1-C n For example, the term "C1-C6" alkyl refers to a moiety having 1-6 carbon atoms. Therefore, "C1-C6 alkyl" refers to an alkyl group having 1-6 carbon atoms, i.e., the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl, etc. The term "alkyl" herein refers to an optionally substituted straight-chain or optionally substituted branched aliphatic hydrocarbon.

[0122] As used herein, “alkyl” includes alkyl groups combined with other groups, such as alkyl groups in alkoxy groups. The term “alkoxy” as used alone or in combination herein refers to an alkyl ether group (O-alkyl), and non-limiting examples of alkoxy groups include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy, etc.

[0123] The term "alkoxycarbonyl" as used alone or in combination in this article refers to alkoxycarbonyl, i.e., -C(O)-O-alkyl.

[0124] The term "aryl / aromatic ring," used alone or in combination herein, refers to an optionally substituted aromatic hydrocarbon group having 6 to about 20, such as 6 to 12 or 6 to 10 cyclic carbon atoms. It can be a fused aromatic ring or a non-fused aromatic ring. A fused aromatic ring comprises 2 to 4 rings, wherein the other independent ring fused with the aryl group can be an alicyclic, heterocyclic, aromatic, or heteroaromatic ring. The aryl group in this document includes monocyclic, bicyclic, tricyclic, or more cyclic aryl groups. Non-limiting examples of monocyclic aryl groups include monocyclic aryl groups with 6 to about 12, 6 to about 10, or 6 to about 8 cyclic carbon atoms, such as phenyl; bicyclic, tricyclic, or more cyclic aryl groups are, for example, naphthyl, phenanthryl, anthracene, or azulel. A non-fused aromatic ring is, for example, a bicyclic non-fused aromatic ring, such as biphenyl.

[0125] The term "heteroaryl / heteroaryl ring" as used alone or in combination herein refers to an arbitrary substituted monovalent heteroaryl group comprising about 5 to about 20, such as 5 to 12 or 5 to 10 skeletal cyclic atoms, wherein one or more (e.g., 1-4, 1-3, 1-2) of the cyclic atoms are heteroatoms, which are independently selected from, but not limited to, heteroatoms of oxygen, nitrogen, and sulfur. The ring of the group does not contain two adjacent O or S atoms. Heteroaryl groups include monocyclic or polycyclic heteroaryl groups (e.g., bicyclic, tricyclic, etc.). In embodiments where two or more heteroatoms are present in the ring, the two or more heteroatoms may be identical to each other, or some or all of the two or more heteroatoms may be different from each other. The term heteroaryl includes an optionally substituted monovalent fused or non-fused heteroaryl group having at least one heteroatom. Fused heteroaryl groups may comprise 2-4 rings, wherein the other rings fused with the heteroaryl group may be alicyclic, heterocyclic, aromatic, or heteroaryl rings. Non-limiting embodiments of monocyclic heteroaryl groups include monocyclic heteroaryl groups with 5 to 12, 5 to 10, 5 to 7, or 6 cyclic skeletal atoms, for example, a non-limiting embodiment includes pyridyl. Fused heteroaryl groups include, for example, benzimidazolyl, quinolinyl, and acridineyl. Non-fused diheteroaryl groups include, for example, dipyridyl. Other embodiments of heteroaryl groups include their oxides, such as pyridyl-N-oxide, etc.

[0126] The term "heterocycle" or "heterocyclic group" as used alone or in combination herein refers to a non-aromatic heterocycle, including heterocyclic alkyl (saturated heterocyclic groups) and heterocyclic alkenyl (unsaturated heterocyclic groups). One or more of the cyclic atoms (e.g., 1-4, 1-3, 1-2) are heteroatoms, such as oxygen, nitrogen, or sulfur atoms. Heterocyclic groups can include monocyclic heterocyclic groups (heterocyclic groups having one ring) or polycyclic heterocyclic groups (e.g., bicyclic heterocyclic groups (heterocyclic groups having two rings), tricyclic heterocyclic groups, etc.). Bicyclic heterocyclic groups can be spirocyclic or bridged rings. Heterocyclic groups can have 3 to about 20, such as 3 to about 10, 3 to about 8, 5 to about 8, or 5 to about 6 cyclic atoms. Heterocyclic groups also include heterocycles with one or more fused aromatic rings (i.e., sharing a common bond), such as 2,3-dihydrobenzofuran, 1,3-benzodioxane, benzo-1,4-dioxane, phthalimide, and naphthalenedioximide. Heterocyclic groups with one or more fused aromatic rings can be linked to other groups via the aromatic or non-aromatic ring portions. Other groups can be linked to the heterocycle via heteroatoms or carbon atoms (i.e., the heterocycle is linked to the parent molecule or further substituted).

[0127] The term "-O-heterocyclic atom" as used alone or in combination in this article refers to an oxygen heterocyclic atom, that is, a heterocyclic atom containing an oxygen atom as a cyclic atom.

[0128] The term "cycloalkyl" as used alone or in combination herein refers to a saturated carbon ring. A cycloalkyl group can be monocyclic or polycyclic (e.g., having 2, 3, or 4 rings), can be spirocyclic or bridged, and can have 3 to 20 carbon atoms, for example, 3 to about 15 cyclic carbon atoms, 3 to about 10 cyclic carbon atoms, or 3 to 6 cyclic carbon atoms. Cycloalkyl groups also include rings having one or more aromatic rings fused together (i.e., sharing a common bond), such as benzopentane, hexane, etc. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.

[0129] The term "substituted" means that one or more hydrogen atoms on a particular atom are replaced by a specified group. If the normal valence of the specified atom is not exceeded under the existing conditions, the result of the substitution is a stable compound.

[0130] Halogens include fluorine, chlorine, bromine, and iodine. A cyano group is represented by "-CN"; a hydroxyl group by "-OH"; a mercapto group by "-SH"; and an amino group by "-NH2".

[0131] As used alone or in combination herein, the terms "halogenated" or "halogen substituted" refer to the replacement of one or more hydrogen atoms in an optionally substituted group (such as an optionally substituted alkyl or alkoxy group) with fluorine, chlorine, bromine, iodine atoms, or combinations thereof. In some embodiments, two or more hydrogen atoms are replaced with the same halogen atom (e.g., difluoromethyl, trifluoromethyl); in other embodiments, two or more hydrogen atoms are replaced with halogen atoms that are not exactly the same (e.g., 1-chloro-1-fluoro-1-iodoethyl). Non-limiting examples of halogenated alkyl groups include trifluoromethyl.

[0132] The term "membered ring" refers to the number of skeleton atoms that make up the ring. For example, pyridine is a six-membered ring, and pyrrole is a five-membered ring.

[0133] As used herein, the term "isotope derivative" refers to a compound in which one or more atoms are replaced by an atom of a different atomic mass or mass number. Isotopes in the compounds listed in this application include H, C, N, and O, such as... 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O.

[0134] As used herein, the terms “subject,” “patient,” or “individual” refer to an individual suffering from a disease, symptom, or condition, including both 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, and pigs; domesticated animals such as rabbits, dogs, and cats; and laboratory animals, including rodents such as rats, mice, and guinea pigs. Examples of non-human mammals include, but are not limited to, birds and fish. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0135] As used herein, the term "treatment" and other similar synonyms include relieving, reducing, or improving symptoms of a disease or condition; inhibiting a disease or condition, such as preventing its progression; alleviating a disease or condition; improving a disease or condition; relieving symptoms caused by a disease or condition; or stopping symptoms of a disease or condition; preventing other symptoms; improving or preventing the underlying metabolic causes of symptoms; and, moreover, the term includes preventative purposes. The term also includes achieving therapeutic and / or preventative effects. A therapeutic effect refers to the cure or improvement of the underlying disease being treated. Furthermore, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect; for example, an improvement is observed in a patient even though they may still be affected by the underlying disease. In terms of preventative effects, the composition may be administered to patients at risk of developing a specific disease, or to patients exhibiting one or more physiological symptoms of a disease, even if no disease diagnosis has been made.

[0136] As used herein, the terms "effective amount," "therapeutic effective amount," or "pharmaceutical effective amount" refer to the amount of at least one active substance (such as the compounds of this application) that, when taken orally, is sufficient to alleviate to some extent one or more symptoms of the disease or condition being treated. The result may be a reduction and / or relief of signs, symptoms, or causes, or any other desired change in the biological system. For example, an "effective amount" for treatment is the amount of a composition comprising the compounds disclosed herein that is clinically necessary to provide significant symptom relief. Effective amounts suitable for any individual case can be determined using techniques such as dose escalation testing.

[0137] As used herein, the terms “administration,” “application,” “dosage,” etc., refer to methods that deliver a compound or composition to the desired site for biological action. These methods include, but are not limited to, oral, duodenal, parenteral (including intravenous, subcutaneous, intraperitoneal, intramuscular, intra-arterial injection or infusion), topical, and rectal administration. Those skilled in the art are familiar with administration techniques that can be used with the compounds and methods described herein, such as those discussed in Goodman and Gilman, *The Pharmacological Basis of Therapeutics*, current ed.; Pergamon; and Remington's, *Pharmaceutical Sciences* (current edition), Mack Publishing Co., Easton, Pa.

[0138] As used herein, the term "pharmaceutical acceptable" means a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of this application and is relatively non-toxic, i.e., that the substance can be administered to an individual without causing an adverse biological reaction or interacting adversely with any component contained in the composition.

[0139] As used herein, the term "pharmaceutical composition" refers to a mixture of the compound of this application with at least one pharmaceutically acceptable substance. The pharmaceutically acceptable substance includes, but is not limited to, a carrier. As used herein, the term "carrier" refers to a relatively non-toxic substance that facilitates the introduction of the compound of this application into cells or tissues.

[0140] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological potency of the free acid and free base of the specified compound and has no adverse effects in biological or other respects. The compounds in this application also include pharmaceutically acceptable salts. A pharmaceutically acceptable salt is one in which a base group in the parent compound is converted into a salt form. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts containing base groups such as amine (amino) groups. Pharmaceutically acceptable salts of this application can be synthesized from the parent compound by reacting a basic group in the parent compound with 1-4 equivalents of an acid in a solvent system. Suitable salts are listed in Remingtong's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977).

[0141] Unless otherwise specified, the term "salt" in this application refers to acidic salts formed from organic / inorganic acids and basic salts formed from organic / inorganic bases. Additionally, when the basic functional group of a compound of the general formula is pyridine or imidazole (but not limited to pyridine or imidazole) and the acidic functional group is a carboxylic acid (but not limited to carboxylic acids), an zwitterion (internal salt) is formed, and internal salts are also included in the salts used in this application.

[0142] The stereoisomers described in this application, unless otherwise stated, include enantiomers and diastereomers, wherein diastereomers further include cis-trans isomers (geometric isomers) and conformational isomers. The compounds containing asymmetrically substituted carbon atoms of this application can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from racemic mixtures or synthesized using chiral starting materials or chiral reagents. The compounds of this application also include tautomer forms. Tautomer forms arise from the exchange of a single bond with an adjacent double bond, accompanied by the migration of a proton. Detailed Implementation

[0143] Exemplary implementations utilizing the principles of this application are described in detail below. A better understanding of the features and advantages of this application can be achieved by referring to the following description.

[0144] The compounds disclosed herein can be prepared by the following general methods. The following general methods and examples are for illustrative purposes only. These methods and examples should not be construed as limiting the scope of this disclosure in any way. The compounds described herein can also be synthesized using standard synthetic techniques known to those skilled in the art, or in combination with methods known in the art.

[0145] The chemical reactions in the embodiments of this disclosure are carried out in a suitable solvent, which must be suitable for the chemical changes of this disclosure and the reagents and materials required therefor. To obtain the compounds of this disclosure, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0146] The reactions described herein can be monitored using any suitable method known in the art. For example, they can be monitored using broad-spectrum methods such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13 C) Infrared spectroscopy, spectrophotometry (e.g., UV-Vis), mass spectrometry, or chromatographic methods such as high performance liquid chromatography (HPLC) or thin-layer chromatography (TLC) to monitor product formation.

[0147] While preferred embodiments of this application are described herein, these embodiments are provided by way of example only. It should be understood that variations of the embodiments described herein can also be used to implement this application. Those skilled in the art will understand that various variations, changes, and substitutions may occur without departing from the scope of this application. It should be understood that the scope of protection of each aspect of this application is determined by the claims, and the methods and structures within the scope of these claims, as well as their equivalents, are all within the scope of these claims.

[0148] Preparation of Formula I compounds

[0149] I. General Synthetic Route I:

[0150] Step 1: Dissolve R-1 and R-2 in a polar solvent (such as N,N-dimethylformamide, dimethyl sulfoxide) and allow them to undergo a docking reaction in the presence of an organic base (triethylamine, diisopropylethylamine, etc.). After the reaction is complete, pour the reaction solution into water to precipitate the crude product. The crude product can be purified by a non-polar solvent such as petroleum ether, or by silica gel column chromatography to obtain intermediate R-3.

[0151] Step 2: Add R-3 to a suitable solvent. Palladium carbon hydrogenation, zinc powder and ammonium acetate reduction, or Raney nickel and hydrazine hydrate can be used to convert the nitro group on the pyridine ring to an amino group. After the reaction is complete, filter the system to remove the catalyst, wash to remove the inorganic salt, and concentrate to obtain intermediate R-4. The product can be directly added to the next reaction or purified for multiple uses.

[0152] Step 3: A suitable solvent can be added to R-4. The amino groups at the 2 and 3 positions of pyridine can be cyclically closed using (alkyl trimethyl ester or triethyl ester, cyanogen bromide, etc.) to obtain a pyridine-imidazole compound. After the reaction is complete, the reaction system is concentrated and extracted to obtain the crude product. Then, the product is purified by purification or column chromatography to obtain compound R-5.

[0153] Step 4: Add a suitable solvent to R-5, then add an acid to remove the two amino protecting groups of R-5. After the reaction is complete, concentrate the solvent, then use a base to convert the salt of R-6 into a free state. Finally, obtain R-6 by crystallization or column chromatography.

[0154] Step 5: Add a suitable solvent to R-6, and then use an amino-binding reagent such as triethyl orthoformate or trimethyl orthoacetate to form an amidine structure from the two amino groups. After the reaction is complete, extract and wash, and then obtain compound R-7 by chromatographic preparation or silica gel chromatography.

[0155] II: General Synthetic Route

[0156] Step 1: Add a suitable polar solvent (DMF / DMSO) to S-1 and S-2. Under alkaline conditions (potassium carbonate, triethylamine, diisopropylethylamine), heat to 50-120℃ and react for 1-4 hours. The amino group of S-1 reacts with the 6-position of S-2. After the reaction is complete, add water to the system. The product precipitates out. Filter to obtain a lower purity product. Then, obtain S-3 by crystallization or silica gel chromatography.

[0157] Step 2: Add S-3 to a suitable solvent (methanol / ethanol / tetrahydrofuran). At room temperature, use a reducing agent (ferric acid / palladium carbon hydrogen / zinc acetic acid / Raney nickel hydrazine hydrate) to convert the nitro group on the pyridine ring to an amino group. After the reaction is complete, filter the system to remove the catalyst, wash to remove inorganic salts, and concentrate to obtain intermediate S-4. The product can be directly added to the next reaction or purified for multiple uses.

[0158] Step 3: Add a suitable solvent (such as alkyl trimethyl ester, triethyl ester, or cyanogen bromide) to S-4 to perform a ring-closure reaction on the amino groups at the 2 and 3 positions of pyridine, to obtain a pyridine-imidazolium compound. After the reaction is complete, concentrate the reaction system, extract the crude product, and then purify it by purification or column chromatography to obtain compound S-5.

[0159] Step 4: Add a suitable solvent (MeOH, THF, DCM) to S-5, and hydrolyze the methoxy group to a hydroxyl group at room temperature by adding an acid or boron tribromide solution. After the reaction is complete, quench the reaction with water, extract with an organic solvent, and dry the product to obtain compound S-6 by crystallization or silica gel chromatography.

[0160] Step 5: Add a suitable solvent (DMF / THF) to S-6, dehydrogenate the imidazole amino hydrogen in a strong base (LiHMDS / NaH), then add a hydroxylamine compound for substitution reaction. After the reaction is complete, add ethyl acetate solvent, wash the organic phase with water to remove the base and residual hydroxylamine, dry the organic phase, filter and concentrate to obtain the crude product, and obtain compound S-7 by crystallization or silica gel chromatography.

[0161] Step 6: Triethyl orthoformate or trimethyl orthoacetate solvent can be added to S-7, and the temperature is raised to 50-120℃ to carry out the reaction. After the reaction is complete, the remaining solvent is removed by direct concentration. The concentrate is then crystallized using a non-polar solvent and filtered to obtain product S-8.

[0162] Step 7: Add a suitable solvent (MeOH, EtOH) to S-8, then add an amino alcohol compound, heat to 30-70℃ to react. After the reaction is complete, concentrate to remove the solvent, then add dichloromethane, wash the organic phase to remove excess amino alcohol, dry the organic phase, filter and concentrate, and purify the concentrated residue using silica gel chromatography to obtain S-9.

[0163] Step 8: Add a suitable solvent (DCM / THF) to S-9, add triphenylphosphine, cool to about 0°C, add DiAD dropwise, and carry out the Mitsunobu reaction. The two hydroxyl groups dock to obtain a macrocyclic compound. After the reaction is complete, concentrate the system and purify it by chromatography or silica gel chromatography to obtain S-10.

[0164] Preparation of main intermediates

[0165] Preparation of tert-butyl ((S)-2-(2-((S)-1-aminoethyl)-4-fluorophenoxy)propyl)carbamate

[0166] Step 1: Synthesis of (S)-(2-(2-acetyl-4-fluorophenoxy)propyl)tert-butyl carbamate

[0167] 30.8 g (0.2 mol) of 1-(5-fluoro-2-hydroxyphenyl)-1-ethyl ketone, 78.6 g (0.3 mol) of triphenylphosphine, and 52.5 g (0.3 mol) of (R)-(2-hydroxypropyl)carbamate tert-butyl ester were dissolved in 300 mL of dichloromethane. The mixture was cooled to 0 °C, and 60.3 g (0.3 mol) of diisopropyl azodicarbonate was slowly added dropwise. After the addition was complete, the reaction was continued for 1 h. TLC showed that the starting material had basically disappeared. 500 mL of petroleum ether was added, and the mixture was stirred to crystallize. The solid was filtered, and the liquid was washed with water and twice with saturated sodium chloride. The liquid was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The oily substance was chromatographically analyzed with an eluent (petroleum ether: ethyl acetate = 20:1-15:1) to obtain 52.3 g of (S)-(2-(2-acetyl-4-fluorophenoxy)propyl)carbamate tert-butyl ester, with a yield of 84.0%.

[0168] Step 2: Synthesis of tert-butyl carbamate ((S)-2-(2-((Z)-1-(((R)-tert-butylsulfinyl)imino)ethyl)-4-fluorophenoxy)propyl)carbamate

[0169] 52.3 g (0.168 mol) of (S)-(2-(2-acetyl-4-fluorophenoxy)propyl)carbamate tert-butyl ester, 76.6 g (0.336 mol) of tetraethoxytitanium, and 30.2 g (0.252 mol) of (R)-2-methylpropane-2-sulfinamide were added to 500 mL of anhydrous tetrahydrofuran. The mixture was heated to 60-65 °C and reacted for 3-4 hours. HPLC showed that the starting material had basically disappeared. The mixture was cooled to room temperature, and water was slowly added dropwise to precipitate a solid. The solid was filtered and washed once with ethyl acetate. The filtrates and ethyl acetate were combined, and the organic phase was washed once with saturated NaHCO3 and twice with saturated NaCl solution. The mixture was concentrated to obtain 72.3 g of crude ((S)-2-(2-((Z)-1-(((R)-tert-butylsulfinyl)imino)ethyl)-4-fluorophenoxy)propyl)carbamate.

[0170] Step 3: Synthesis of tert-butyl carbamate ((S)-2-(2-((S)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate

[0171] 72.3 g (0.168 mol) of tert-butyl carbamate ((S)-2-(2-((Z)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate was dissolved in 95% methanol aqueous solution. The solution was cooled to -30 to -50 °C, and kept below -30 °C. 6.3 g (0.168 mol) of sodium borohydride was added in portions. After the addition was complete, the reaction was continued for 1 hour. After the reaction was complete, water and ethyl acetate were added, and the mixture was stirred and separated. The organic phase was washed twice with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to chromatography using petroleum ether:ethyl acetate = 20:1 to 5:1 to obtain 60.4 g of tert-butyl carbamate ((S)-2-(2-((S)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate. The two-step yield was 86.3%.

[0172] Step 4: Synthesis of tert-butyl ((S)-2-(2-((S)-1-aminoethyl)-4-fluorophenoxy)propyl)carbamate

[0173] 60.4 g (0.145 mol) of tert-butyl carbamate ((S)-2-(2-((S)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate tert-butyl ester was dissolved in 500 mL of 80% methanol aqueous solution, and 9.2 g (0.036 mol) of elemental iodine was added. The reaction was carried out at room temperature for 3 h, and the color of the system changed from brown to pale yellow. TLC showed that the reactants had completely reacted. The solvent was removed by concentration, and 400 mL of water was added to make... Extracted twice with petroleum ether:ethyl acetate at a ratio of 10:1, the aqueous phase was then extracted twice more with ethyl acetate. The ethyl acetate phase was washed twice with saturated NaHCO3 solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain an oily substance. Chromatography using dichloromethane:methanol at a ratio of 50:1 to 20:1 yielded 27.2 g of the product ((S)-2-(2-((S)-1-aminoethyl)-4-fluorophenoxy)propyl)carbamate, with a yield of 60.0%. 1 H NMR(400MHz, CDCl3)δ7.00(dd,J=9.5,2.8Hz,1H),6.83–6.70(m,2H),5.67(s,1H),4.36(dd,J=30.7,4 .8Hz,2H),3.36(s,1H),3.23–3.13(m,1H),1.34(s,9H),1.30(d,J=6.7Hz,3H),1.22(d,J=6.1Hz,3H).

[0174] Preparation of N-(2-bromo-3-fluoro-6-methoxybenzyl)ethylamine

[0175] 22.1 g (0.1 mol) of 2-bromo-3,6-difluorobenzaldehyde and 5.72 g (0.11 mol) of sodium methoxide were dissolved in 200 mL of methanol. The mixture was heated to 55-60 °C and reacted for 6-8 h under TLC monitoring. When a small amount of raw material remained, the reaction was stopped, the methanol was removed by concentration, ethyl acetate was added, and the organic phase was washed twice with saturated NaCl solution. The phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to chromatography using an eluent (petroleum ether:ethyl acetate = 20:1 to 10:1) to obtain 18.9 g of 2-bromo-3-fluoro-6-methoxybenzaldehyde, with a yield of 81.1%.

[0176] Step 2: Synthesis of N-(2-bromo-3-fluoro-6-methoxybenzyl)ethylamine

[0177] 18.9 g (0.081 mol) of 2-bromo-3-fluoro-6-methoxybenzaldehyde was dissolved in 200 mL of methanol, and 6.7 g (0.104 mol) of 70% ethylamine aqueous solution was added. The mixture was heated to 50 °C and reacted for 1 hour. After the reaction was completed, the temperature was lowered to 0–5 °C, and 3.0 g (0.081 mL) of sodium borohydride solid was added in portions. After the addition was completed, the reaction was carried out for 30 min. The solvent was removed by concentration, and the remaining oily substance was added to ethyl acetate. The mixture was washed once with saturated NaHCO3 and twice with saturated NaCl. The solution was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The oily substance was then subjected to chromatography using an eluent (petroleum ether:ethyl acetate 5:1–3:1) to obtain 19.7 g of N-(2-bromo-3-fluoro-6-methoxybenzyl)ethylamine, with a yield of 92.9%.

[0178] Preparation of N-(5-fluoro-2-methoxybenzyl)ethylamine

[0179] 20 g (0.13 mol) of 5-fluoro-2-methoxybenzaldehyde and 10.0 g (0.156 mol) of 70% ethylamine aqueous solution were dissolved in 200 mL of methanol. The mixture was heated to 50 °C and reacted for 1 hour. After the reaction was complete, the temperature was lowered to 0–5 °C, and 4.8 g (0.13 mol) of sodium borohydride solid was added in portions. After the addition was complete, the reaction was carried out for 30 min. The solvent was removed by concentration, and the remaining oily substance was added to ethyl acetate. The mixture was washed once with saturated NaHCO3 and twice with saturated NaCl. The solution was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain an oily substance. The product was obtained by chromatography using a petroleum ether:ethyl acetate 5:1–3:1 eluent, yielding 19.4 g of N-(5-fluoro-2-methoxybenzyl)ethylamine, with a yield of 87.8%.

[0180] N-(2-bromo-6-methoxybenzyl)ethylamine; 2-((ethylamino)methyl)-3-methoxybenzonitrile; 2-((ethylamino)methyl)-6-fluoro-3-methoxybenzonitrile were prepared using similar methods.

[0181] Example A-1: ​​Preparation of (3S,6S,E)-4 6 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,3)-benzocyclodecane-9-ene

[0182] Step 1: Synthesis of tert-butyl 2-(6-(((S)-1-(2-((((S)-1-((tert-Butoxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylic acid

[0183] 17.0 g (54.5 mmol) of tert-butyl ((S)-2-(2-((S)-1-aminoethyl)-4-fluorophenoxy)propyl)carbamate was dissolved in 250 mL of dry DMF. Then, 14.90 g (51.7 mol) of tert-butyl 2-(6-chloro-3-nitropyridin-2-yl)hydrazide-1-carboxylate and 21.0 g (163.0 mol) of diisopropylethylamine were added at 0 °C. The mixture was heated to 55 °C and reacted for 16 h. TLC showed that the reaction was complete. The reaction solution was washed twice with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (PE / EA = 30 / 1 to 10 / 1, V / V) to give 27.3 g of tert-butyl 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylate, with a yield of 93.6%.

[0184] Step 2: Synthesis of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butoxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid

[0185] 27.3 g (48.0 mol) of 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylic acid tert-butyl ester dissolved in oxygen-free THF was added to a reaction flask. 8.2 g of Raney-Ni catalyst was added under nitrogen protection, followed by the slow addition of 5.3 g (132.0 mmol) of 80% hydrazine hydrate at room temperature. After the addition was complete, the reaction mixture was stirred; the bubbles were colorless, and the system was a transparent liquid. HPLC analysis confirmed the reaction was complete. The reaction mixture was filtered quickly through a diatomaceous earth filter under a nitrogen atmosphere. The filtrate was concentrated to dryness under reduced pressure to obtain 26.7 g of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid. The crude product rapidly darkened in color and was used directly in the next reaction without further purification.

[0186] Step 3: Synthesis of tert-butyl carbamate ((2S)-2-(2-(1-((3-((tert-butoxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate

[0187] 26.7 g of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylate was dissolved in 100 mL of oxygen-free 1,4-dioxane and 100 mL of triethyl orthoformate under nitrogen protection. 10.0 mL of acetic acid was added, and the mixture was heated to 75–80 °C and allowed to rest for 2–3 hours. HPLC showed that the reaction proceeded completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of ethyl acetate, washed twice with organic phase NaHCO3, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 5:1–3:1) yielded 18.2 g of a white solid (((2S)-2-(2-(1-((3-((tert-butyloxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate tert-butyl ester. The two-step yield was 69.0%.

[0188] Step 4: N 5 Synthesis of -((S)-1-(2-(((S)-1-aminopropyl-2-yl)oxy)-5-fluorophenyl)ethyl)-3H-imidazo[4,5-b]pyridine-3,5-diamine

[0189] 18.2 g (33.3 mmol) of tert-butyl carbamate ((2S)-2-(2-(1-((3-((tert-butyloxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate was dissolved in 200 mL of 1 M hydrochloric acid-ethanol solution. The mixture was stirred at room temperature for 1–2 hours, and HPLC showed complete reaction. The reaction solution was concentrated under reduced pressure. Sodium bicarbonate solid was added to the remaining solution in a 2:1 mixture of dichloromethane and methanol, stirred for 10 minutes, filtered, and the liquid was concentrated to dryness to obtain a white solid N. 5 -((S)-1-(2-(((S)-1-aminopropyl-2-yl)oxy)-5-fluorophenyl)ethyl)-3H-imidazo[4,5-b]pyridine-3,5-diamine 11.73 g, yield 84.4%.

[0190] Step 5: (3S, 6S, Z) - 4 5 -Fluoro-3,6-dimethyl-1 3 Synthesis of H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0191] N 5 100 mg (0.29 mmol) of -((S)-1-(2-(((S)-1-aminopropyl-2-yl)oxy)-5-fluorophenyl)ethyl)-3H-imidazo[4,5-b]pyridine-3,5-diamine was dissolved in 5.0 mL of anoxic 1,4-dioxane solution under nitrogen protection, along with 128 mg (0.87 mmol) of triethyl orthoformate. 52 mg (0.87 mmol) of acetic acid was added, and the mixture was heated to 75–80 °C for 2–3 hours. HPLC showed complete reaction of the starting material. The solvent was removed by concentration under reduced pressure. The residue was dissolved in a suitable amount of dichloromethane, washed twice with NaHCO3, once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was then subjected to column chromatography (DCM:MeOH = 50:1–10:1) to obtain a white solid (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 86.0 mg of H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene, yield 83.4%. 1H NMR(400MHz,DMSO-d6)δ7.77(s,1H),7.62–7.59(m,1H),7.41(s,1H),7.16(d t,J=8.4,4.2Hz,1H),6.95–6.88(m,2H),6.41(d,J=8.7Hz,1H),5.76(s,1H),4 .68–4.56(m,1H),3.47(dd,J=14.9,5.7Hz,1H),3.31–3.19(m,1H),2.51(d,J =1.5Hz,1H),1.46–1.35(m,2H),1.32(d,J=7.0Hz,3H),1.28(d,J=6.2Hz,3H). MS: 355.4[M+H] + .

[0192] Example A-2: Preparation of (3S,6S,Z)-4 5 -Fluoro-12,3,6-trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0193] Step 1: Synthesis of tert-butyl carbamate ((2S)-2-(2-(1-((3-((tert-butyloxycarbonyl)amino)-2-methyl-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate

[0194] 5.8 g (2.0 mmol) of 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid tert-butyl ester, 6.5 g (4.0 mmol) of triethyl orthoacetate, and 1.8 g (3.0 mmol) of acetic acid were added to 50 mL of 1,4-dioxane. The mixture was heated to 70-75 °C and reacted for 2-3 hours. After the reaction was complete, the mixture was concentrated to remove the waste. Solvent was used; dichloromethane was added, and the system was washed twice with NaHCO3 solution and twice with NaCl solution. After drying, the crude product was obtained by filtration and concentration. Column chromatography (PE:EA = 5:1 to 3:1) was performed to obtain 4.87 g of a white solid ((2S)-2-(2-(1-((3-((tert-butoxycarbonyl)amino)-2-methyl-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate compound, yield 77.5%).

[0195] The second and third steps are the same as the fourth and fifth steps in Example 1, ultimately yielding compound (3S,6S,Z)-4. 5-Fluoro-12,3,6-trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene, 1 H NMR(400MHz, DMSO-d6)δ8.80(d,J=12.3Hz,1H),7.46(dd,J=10.4,5.1Hz,2H),7.28–7.14(m,2H),6.89(ddd,J=1 1.1,8.7,3.9Hz,2H),6.33(d,J=8.6Hz,1H),5.53(s,1H),4.60(s,1H),2.31(d,J=9.0Hz,3H),1.39-1.21(m,9H). MS:408.3[M+H] + .

[0196] Example A-3 Preparation of (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine

[0197] Step 1: Synthesis of tert-butyl carbamate ((2S)-2-(2-(1-((2-amino-3-((tert-butoxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate

[0198] 2.0 g (3.75 mmol) of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylate was dissolved in 20 mL of dichloromethane under nitrogen protection, and 0.48 g (4.50 mmol) of cyanogen bromide was added. The mixture was stirred at room temperature for 16 hours, and HPLC showed that the reaction proceeded completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of ethyl acetate, washed twice with organic phase NaHCO3, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 5:1–3:1) yielded 1.36 g of a brown solid ((2S)-2-(2-(1-((2-amino-3-((tert-butoxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenoxy)propyl)carbamate tert-butyl ester. Yield: 65.0%.

[0199] The second and third steps are performed following the experimental procedures in steps four and five of Example 1, yielding a white solid (3S,6S,Z)-4. 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine. 1 H NMR (400MHz, DMSO-d6) δ8.92(s,1H),7.27–7.09(m,2H),7.01–6.79(m,3H),6.29(d,J=8.4Hz,1H),6.16(d,J=8. 3Hz,1H),5.95(s,1H),5.83(s,1H),5.41(d,J=74.3Hz,1H),4.63–4.41(m,1H),1.92(s,2H),1.42–1.23(m,6H). MS:370.4[M+H] + .

[0200] Example A-4: Preparation of (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 methyl formate

[0201] Step 1: Synthesis of methyl 3-((tert-Butoxycarbonyl)amino)-5-((1-(2-(((S)-1-((tert-Butoxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-3H-imidazo[4,5-b]pyridine-2-carboxylate

[0202] 2.0 g (3.75 mmol) of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylate was dissolved in anhydrous methanol under nitrogen protection. 0.77 g (4.5 mmol) of methyl 2,2-dichloro-2-methoxyacetate and 1.5 g (15.0 mmol) of triethylamine were added. The mixture was heated to 50–55 °C and reacted for 2–4 hours. HPLC showed that the reaction proceeded completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of ethyl acetate, washed twice with organic phase NaHCO3, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 5:1–3:1) yielded 1.64 g of a white solid, methyl 3-((tert-butoxycarbonyl)amino)-5-((1-(2-(((S)-1-((tert-butoxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-3H-imidazo[4,5-b]pyridine-2-carboxylate. Yield: 72.6%.

[0203] The second and third steps are the same as the fourth and fifth steps in Example 1, yielding a white solid (3S,6S,Z)-4. 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 methyl formate. 1 H NMR (400MHz, DMSO-d6) δ7.78–7.46(m,2H),7.27(d,J=11.4Hz,1H),6.98(dtdd,J=22.8,16.7,11.3,7.3Hz,3H),6.52(dd,J=22.2,8.5Hz,1H),5.6 9–5.30(m,1H),4.81(dd,J=78.0,5.7Hz,1H),4.23(s,1H),3.86(s,3H), 3.52(dd,J=13.3,6.4Hz,1H), 3.17(t,J=13.8Hz,1H), 1.48–1.20(m,6H). MS:413[M+H] + .

[0204] Example A-5

[0205] Method 2: Preparation of (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0206] Step 1: N 2 -Ethyl-N 2 Synthesis of -(5-fluoro-2-methoxybenzyl)-5-nitropyridine-2,6-diamine

[0207] 15.37 g (0.088 mol) of 6-chloro-3-nitropyridine-2-amine and 18.24 g (0.088 mol) of N-(5-fluoro-2-methoxybenzyl)ethylamine were dissolved in anhydrous DMF. 35.50 g (0.275 mol) of diisopropylethylamine was added first, and the mixture was heated at 60 °C under nitrogen protection. ～ The reaction was carried out at 120℃ for 2.5 hours. After the reaction was complete as detected by TLC, the reaction mixture was cooled to room temperature and slowly poured into ice water. The mixture was extracted three times with ethyl acetate. The extracted organic phases were combined, washed once with saturated brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography using EA:PE = 1:10–1:5 as the eluent. Column chromatography with EA:PE = 1:10–1:5 yielded an orange-yellow foamy solid product N. 2 -Ethyl-N 2 22.71 g of -(5-fluoro-2-methoxybenzyl)-5-nitropyridine-2,6-diamine, yield 83.6%.

[0208] Step 2: N 6 -Ethyl-N 6 Synthesis of 5-fluoro-2-methoxybenzyl)pyridine-2,3,6-triamine

[0209] N 2 -Ethyl-N 2 22.42 g (0.07 mol) of 5-fluoro-2-methoxybenzyl)-5-nitropyridine-2,6-diamine was dissolved in anhydrous THF, and 11.25 g of Raney nickel was added. Then, 14.00 g of hydrazine hydrate was added dropwise at room temperature. After the addition was complete, the reaction continued for 20 min until the system became colorless and clear. The mixture was rapidly filtered, and the filtrate was concentrated to dryness to obtain foamy solid N. 6 -Ethyl-N 6 21.79 g of crude (5-fluoro-2-methoxybenzyl)pyridine-2,3,6-triamine was directly added to the next reaction step.

[0210] Step 3: Synthesis of N-ethyl-N-(5-fluoro-2-methoxybenzyl)-3H-imidazo[4,5-b]pyridine-5-amine

[0211] N 6 -Ethyl-N6 21.79 g of 5-fluoro-2-methoxybenzyl)pyridine-2,3,6-triamine, 50 mL of triethyl orthoformate, 5.0 g of acetic acid, and 100 mL of 1,4-dioxane were mixed and heated to 70-75 °C for 2-3 h. After the reaction was complete, the mixture was concentrated to remove all solvents. Dichloromethane was added, and the system was washed twice with NaHCO3 solution and twice with NaCl solution. The mixture was dried, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 5:1 to 3:1) yielded 17.13 g of N-ethyl-N-(5-fluoro-2-methoxybenzyl)-3H-imidazo[4,5-b]pyridine-5-amine, with a two-step yield of 81.6%.

[0212] Step 4: Synthesis of 2-((ethyl(3H-imidazo[4,5-b]pyridin-5-yl)amino)methyl)-4-fluorophenol

[0213] 17.13 g (0.057 mol) of N-ethyl-N-(5-fluoro-2-methoxybenzyl)-3H-imidazo[4,5-b]pyridin-5-amine was dissolved in dichloromethane, cooled to about 0 °C, and boron tribromide solution (1 M, 171 mL) was added dropwise. After the addition was complete, the mixture was allowed to naturally warm to room temperature and reacted for 4-6 h. After the reaction was complete, water was added dropwise to quench the reaction. The system was washed twice with NaHCO3 solution, dried, filtered, and concentrated to obtain the crude product. The crude product was then obtained by column chromatography (DCM:MeOH = 100:1~30:1) and tested white to give 13.88 g of 2-((ethyl(3H-imidazo[4,5-b]pyridin-5-yl)amino)methyl)-4-fluorophenol, with a yield of 85.0%.

[0214] Step 5: Synthesis of 2-(((3-amino-3H-imidazo[4,5-b]pyridin-5-yl)(ethyl)amino)methyl)-4-fluorophenol

[0215] 13.88 g (0.0485 mol) of 2-((ethyl(3H-imidazo[4,5-b]pyridin-5-yl)amino)methyl)-4-fluorophenol was dissolved in DMF, cooled to 0 °C, and 7.76 g (0.194 mol) of 60% NaH was added in portions. After the addition was complete, the reaction was maintained at this temperature for 1 h. After the reaction was complete, 22.6 g (0.097 mol) of diphenylphosphohydroxylamine was added. After the addition was complete, the temperature was naturally raised to room temperature and the reaction was carried out for 3 h. After the reaction was complete, water and ethyl acetate were added, the mixture was separated, the organic phase was washed with a small amount of water, dried, filtered and concentrated, and the solution was purified by column chromatography (DCM:MeOH = 100:1 to 30:1) to obtain 9.49 g of white solid 2-(((3-amino-3H-imidazo[4,5-b]pyridin-5-yl)(ethyl)amino)methyl)-4-fluorophenol, with a yield of 60.5%.

[0216] Step 6: Synthesis of methyl(Z)-N-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)formimide ester

[0217] 301 mg (1.0 mmol) of 2-(((3-amino-3H-imidazo[4,5-b]pyridin-5-yl)(ethyl)amino)methyl)-4-fluorophenol was suspended in 5.0 mL of excess triethyl orthoformate and heated to 80-1200 °C under nitrogen protection with stirring for 2-3 hours. The reaction was confirmed to be complete by TLC. Excess triethyl orthoformate was removed by concentration under reduced pressure. The residue was dissolved in 1,4-dioxane and concentrated to dryness under reduced pressure. The residue was slurried with ethyl acetate / petroleum ether (1 / 10, V / V), the solid was filtered, and dried to give 286 mg of methyl(Z)-N-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)formimide ester, with a yield of 83.4%.

[0218] Step 7: Synthesis of (R,Z)-N'-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-(2-hydroxypropyl)formimide

[0219] 286 mg (0.83 mmol) of methyl (Z)-N-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)formimide ester was dissolved in anhydrous ethanol, and 187 mg (2.5 mmol) of (R)-2-aminopropanol was added. The mixture was heated to 40-70 °C and stirred overnight. The reaction was confirmed to be complete by HPLC. The reaction mixture was concentrated to dryness under reduced pressure. The residue was dissolved in dichloromethane, washed once with pure water or dilute weak acid, and once with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration, and the filtrate was concentrated to the crude product under reduced pressure. The crude product was purified by a rapid column chromatography system (eluent: MeOH / DCM = 1 / 150 to 1 / 50, V / V) to give 245 mg of (R,Z)-N'-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-(2-hydroxypropyl)formimide, with a yield of 76.1%.

[0220] Step 8: (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 Synthesis of H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene:

[0221] 245 mg (0.63 mmol) of (R,Z)-N'-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-(2-hydroxypropyl)formimide ester was dissolved in tetrahydrofuran, and then 3500 mg (1.90 mmol) of PPh was added and stirred until dissolved. Then, 331 mg (1.90 mmol) of DIAD3 was added dropwise using a syringe. After the addition was complete, the reaction mixture was stirred at room temperature for 2–3 hours, and the reaction was confirmed to be complete by TLC. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was purified by rapid column chromatography (eluent: MeOH / DCM = 1 / 50–1 / 20, V / V) to give a white solid ((S,Z)-2-ethyl-4-yl)formimide ester. 5 -Fluoro-6-methyl-1 3 175 mg of H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene, yield 75.0%. 1 H NMR (400MHz, CDCl3) δ8.81 (s, 1H), 7.80 (s, 1H), 7.78 (s, 1H), 6.86 (dd, J = 9.1, 2. 6Hz,1H),6.82(dd,J=8.2,3.1Hz,1H),6.72(d,J=4.8Hz,1H),6.47(d,J=9.0Hz,1H ),4.56(s,2H),3.46(q,J=7.1Hz,2H),2.76–2.67(m,1H),2.64(d,J=6.0Hz,1H), 2.11(d,J=3.7Hz,1H),1.65(s,1H),1.35(d,J=5.5Hz,3H),1.15(t,J=7.1Hz,3H). MS: 369.3 [M+H] + .

[0222] Example A-6 Preparation of (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile

[0223] Step 1: Preparation of 2-(((6-amino-5-nitropyridin-2-yl)(ethyl)amino)methyl)-6-fluoro-3-methoxybenzonitrile

[0224] 5.0 g (24.0 mmol) of 2-((ethylamino)methyl)-6-fluoro-3-methoxybenzonitrile, 4.16 g (24.0 mmol) of 6-chloro-3-nitropyridin-2-amine, and 9.29 g (72.0 mmol) of diisopropylethylamine were added to 30 mL of N,N-dimethylformamide. The reaction was carried out at 50 °C for 2 h, and TLC showed that the reaction was complete. The system was poured into water, and a solid precipitated. The solid was filtered, and dissolved in petroleum ether and ethyl acetate (5:1) by heating. The solid precipitated by cooling and filtration yielded 7.20 g of a pale yellow solid, 2-(((6-amino-5-nitropyridin-2-yl)(ethyl)amino)methyl)-6-fluoro-3-methoxybenzonitrile, in 87.0% yield.

[0225] The second step and subsequent experimental procedures were performed in accordance with the following steps in Example 5, ultimately yielding compound (S,Z)-2-ethyl-4-ethyl. 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -Formonitrile. 1 H NMR (400MHz, CDCl3) δ8.58(s,1H),7.65(s,1H),7.03(dd,J=8.9,4.4Hz,1H),6.94(d,J=8.2Hz,1H),6.84(dd,J=8.9,4.1Hz,1H),6.62(d,J= 9.0Hz,1H),4.51(dd,J=37.0,30.7Hz,2H),3.62–3.51(m,2H),3.50–3.38(m,2H),2.02(s,2H),1.40(d,J=6.2Hz,3H),1.14(t,J=6.9Hz,3H). MS:494.3[M+H] + .

[0226] The compounds in the following table were prepared using the same method:

[0227] Example A-11, (S,Z)-2-ethyl-4 5 -Fluorine-1 2 ,6-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 Preparation of formonitrile

[0228] Step 1: Synthesis of 2-((ethyl(2-methyl-3H-imidazo[4,5-b]pyridin-5-yl)amino)methyl)-6-fluoro-3-methoxybenzonitrile)

[0229] 6.30 g (20.0 mmol) of 2-(((5,6-diaminopyridin-2-yl)(ethyl)amino)methyl)-6-fluoro-3-methoxybenzonitrile, 50 mL of triethyl orthoacetate, 5.0 g of acetic acid, and 50 mL of 1,4-dioxane were mixed and heated to 85 °C for 2-3 h. After the reaction was complete, the mixture was concentrated to remove all solvents. Dichloromethane was added, and the system was washed twice with NaHCO3 solution and twice with NaCl solution. The mixture was dried, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 5:1 to 3:1) yielded 5.15 g of 2-((ethyl(2-methyl-3H-imidazo[4,5-b]pyridin-5-yl)amino)methyl)-6-fluoro-3-methoxybenzonitrile, with a yield of 76.0%.

[0230] Steps two through six are the same as steps four through eight of Example 5, yielding the product (S,Z)-2-ethyl-4-ethyl. 5 -Fluorine-1 2 ,6-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile, 1 H NMR (400MHz, CDCl3) δ9.09 (s, 1H), 8.24 (s, 1H), 7.72 (dd, J = 19.8, 8.7Hz, 1H), 7.11 (dd, J =8.9,4.4Hz,1H),7.01(t,J=8.4Hz,1H),6.92(dd,J=9.0,4.2Hz,1H),4.69–4.55(m,1H),4 .14(dd,J=15.2,7.2Hz,1H),3.75(d,J=15.1Hz,1H),3.60(dd,J=15.4,7.5Hz,1H),3.49- 3.22 (dd, J = 23.3, 11.4 Hz, 3H), 2.43 (s, 3H), 1.33 (d, J = 6.0 Hz, 3H), 1.19 (t, J = 6.9 Hz, 3H). MS:408.3[M+H] + .

[0231] Example A-12: Preparation of (R,Z)-2-ethyl-4,5-fluoro-7-methyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene

[0232] Step 1: Synthesis of (R,Z)-N'-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-(1-hydroxypropyl-2-yl)formimidamide

[0233] 200 mg (0.58 mmol) of methyl (Z)-N-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)formimidate was dissolved in 5.0 mL of anhydrous ethanol, and 87 mg (1.16 mmol) of (R)-2-amino-1-propanol was added. The mixture was heated to 65 °C and stirred for 3 h. The reaction was confirmed to be complete by HPLC. The reaction mixture was concentrated to dryness under reduced pressure. The residue was dissolved in dichloromethane, washed once with pure water or dilute weak acid, and once with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration, and the filtrate was concentrated to the crude product under reduced pressure. The crude product was purified by a rapid column chromatography system (eluent: MeOH / DCM = 1 / 150 to 1 / 50, V / V) to give 164 mg of (R,Z)-N'-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-(1-hydroxypropyl-2-yl)iminoformamide, with a yield of 72.3%.

[0234] Step 2: Synthesis of (R,Z)-2-ethyl-4,5-fluoro-7-methyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzocyclodecane-9-ene

[0235] 164 mg (0.42 mmol) of (R,Z)-N'-(5-(ethyl(5-fluoro-2-hydroxybenzyl)amino)-3H-imidazo[4,5-b]pyridin-3-yl)-N-(1-hydroxypropyl-2-yl)formimide amide was dissolved in tetrahydrofuran, and then 330 mg (1.26 mmol) of PPh3 was added and stirred until dissolved. Then, 220 mg (1.26 mmol) of DIAD2 was added dropwise using a syringe. After the addition was complete, the reaction mixture was stirred at room temperature for 2–3 hours, and the reaction was confirmed to be complete by TLC. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was purified by rapid column chromatography (eluent: MeOH / DCM = 1 / 50 to 1 / 20, V / V) to give 128 mg of a white solid (R,Z)-2-ethyl-4,5-fluoro-7-methyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene, yield 82.6%. 1H NMR (400MHz, CDCl3) δ8.24–7.99(m,1H),7.79(d,J=8.2Hz,1H),7.75(s,1H),6.89(s,1H),6.86(s,1H),6.74(s,1H),6.48(d,J= 8.1Hz,1H),4.56(s,2H),3.48(d,J=16.5Hz,2H),3.37(s,2H),2.70(m,1H),1.80(s,2H),1.63(s,2H),1.38(d.3H),1.19(m,3H). MS:369.3[M+H] + .

[0236] Using similar intermediates, the following compounds can be prepared using the same method:

[0237] Example A-25: Preparation of (R,Z)-2-ethyl-4 5 -Fluorine-1 2 7-Dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile((R,Z)-2-ethyl-4 5 -fluoro-1 2 ,7-dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridina-4(1,2)-benzenacyclodecaphan-9-ene-4 6 -carbonitrile)

[0238] Step 1: Synthesis of (R,Z)-N'-(5-((2-cyano-3-fluoro-6-hydroxybenzyl)(ethyl)amino)-2-methyl-3H-imidazo[4,5-b]pyridin-3-yl)-N-(1-hydroxypropyl-2-yl)formimide amide

[0239] 200 mg (0.52 mmol) of methyl (Z)-N-(5-(((2-cyano-3-fluoro-6-hydroxybenzyl)(ethyl)amino)-2-methyl-3H-imidazo[4,5-b]pyridin-3-yl))formimide ester was dissolved in 5.0 mL of anhydrous ethanol, and 78 mg (1.05 mmol) of (R)-2-amino-1-propanol was added. The mixture was heated to 50 °C and stirred for 4 h. The reaction was confirmed to be complete by HPLC. The reaction mixture was concentrated to dryness under reduced pressure. The residue was dissolved in dichloromethane, washed once with pure water or dilute weak acid, and once with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration, and the filtrate was concentrated to the crude product under reduced pressure. The crude product was purified by a rapid column chromatography system (eluent: MeOH / DCM = 1 / 150 to 1 / 50, V / V) to give 175 mg of (R,Z)-N'-(5-((2-cyano-3-fluoro-6-hydroxybenzyl)(ethyl)amino)-2-methyl-3H-imidazo[4,5-b]pyridin-3-yl)-N-(1-hydroxypropyl-2-yl)formimide amide, with a yield of 78.8%.

[0240] Step 2: (R,Z)-2-ethyl-4 5 -Fluorine-1 2 7-Dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 Synthesis of formonitrile

[0241] 175 mg (0.41 mmol) of (R,Z)-N'-(5-(((2-cyano-3-fluoro-6-hydroxybenzyl)(ethyl)amino)-2-methyl-3H-imidazo[4,5-b]pyridin-3-yl)-N-(1-hydroxypropyl-2-yl)carboximide amide was dissolved in tetrahydrofuran, and then 322 mg (1.23 mmol) of PPh3 was added and stirred until dissolved. Then, 15 mg (1.23 mmol) of DIAD2 was added dropwise using a syringe. After the addition was complete, the reaction mixture was stirred at room temperature for 1 hour, and the reaction was confirmed to be complete by TLC. The reaction mixture was concentrated to dryness under reduced pressure, and the residue was purified by rapid column chromatography (eluent: MeOH / DCM = 1 / 50 to 1 / 20, V / V) to give a white solid (R,Z)-2-ethyl-4-yl 5 -Fluorine-1 2 7-Dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -Formonitrile 135mg, yield 82.6%. 1H NMR(400MHz, CDCl3)δ7.75(dd,J=19.9,8.4Hz,1H),7.35–6.86(m,3H),6.72–6.48(m,1H),4.23–4.03(m,2H),3 .79(d,J=13.9Hz,1H),3.71–3.52(m,3H),2.51(d,J=35.2Hz,3H),2.15(s,2H),1.39(s,3H),1.26–1.19(m,3H). MS:408.3[M+H] + .

[0242] The following compounds can be prepared using the same method:

[0243] Example A-31: (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 Preparation of -amines:

[0244] Step 1: Synthesis of tert-butyl (S)-2-(6-((2-((1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorobenzyl)(ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylic acid

[0245] 20.0 g (61.3 mmol) of (S)-(2-(2-((ethylamino)methyl)-4-fluorophenoxy)propyl)carbamate tert-butyl ester was dissolved in 250 mL of dry DMF. Then, 17.70 g (61.3 mol) of 2-(6-chloro-3-nitropyridin-2-yl)hydrazide-1-carboxylate tert-butyl ester and 23.74 g (184.0 mol) of diisopropylethylamine were added at 0 °C. The mixture was heated to 75 °C and reacted for 4 h. TLC showed that the reaction was complete. The reaction solution was washed twice with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (PE / EA = 30 / 1~10 / 1, V / V) to give 28.7 g of (S)-2-(6-((2-((1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorobenzyl)(ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylic acid tert-butyl ester, with a yield of 81.0%.

[0246] Step 2: Synthesis of tert-butyl (S)-2-(3-amino-6-((2-((1-((tert-butoxycarbonyl)amino)prop-2-yl)oxy)-5-fluorobenzyl)(ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid

[0247] 28.7 g (49.6 mmol) of (S)-2-(6-((2-((1-((tert-butyloxycarbonyl)amino)propyl-2-yl)oxy)-5-fluorobenzyl)(ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylic acid tert-butyl ester was added to a reaction flask in oxygen-free THF. 8.5 g of Raney-Ni catalyst was added under nitrogen protection, followed by the slow addition of 7.5 g (118 mmol) of 80% hydrazine hydrate at room temperature. After the addition was complete, the reaction mixture was stirred; the bubbles were colorless, and the system was a transparent liquid. HPLC analysis confirmed the reaction was complete. The reaction mixture was filtered quickly through a diatomaceous earth filter under nitrogen atmosphere. The filtrate was concentrated to dryness under reduced pressure, yielding 25.7 g of crude (S)-2-(3-amino-6-((2-((1-((tert-butyloxycarbonyl)amino)propyl-2-yl)oxy)-5-fluorobenzyl)(ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid tert-butyl ester. The crude product rapidly darkened in color and was used directly in the next reaction without further purification.

[0248] Step 3: Synthesis of (S)-(2-(2-(((2-amino-3-((tert-butoxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)(ethyl)amino)methyl)-4-fluorophenoxy)propyl)tert-butyl carbamate

[0249] 25.7 g of (S)-2-(3-amino-6-((2-((1-((tert-butoxycarbonyl)amino)propyl-2-yl)oxy)-5-fluorobenzyl)(ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid tert-butyl ester was dissolved in 125 mL of dichloromethane and 5.8 g (56.2 mmol) of cyanogen bromide under nitrogen protection. The reaction was carried out at room temperature for 16 hours. HPLC showed that the starting material reacted completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of ethyl acetate, washed twice with organic phase NaHCO3, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 4:1–2:1) yielded 16.8 g of a white solid (S)-(2-(2-(((2-amino-3-((tert-butoxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)(ethyl)amino)methyl)-4-fluorophenoxy)propyl)carbamate tert-butyl. Two-step yield: 62.6%.

[0250] Step 4: (S)-N 5 -(2-((1-aminopropyl-2-yl)oxy)-5-fluorobenzyl)-N 5Synthesis of ethyl-3H-imidazo[4,5-b]pyridine-2,3,5-triamine

[0251] 16.8 g (29.3 mmol) of (S)-(2-(2-(((2-amino-3-((tert-butoxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)(ethyl)amino)methyl)-4-fluorophenoxy)propyl)carbamate tert-butyl ester was dissolved in 200 mL of 1 M hydrochloric acid-ethanol solution. The mixture was stirred at room temperature for 2 hours, and HPLC showed complete reaction. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in a solution of dichloromethane and methanol (2:1). Sodium bicarbonate solid was added, and the mixture was stirred for 10 minutes. The mixture was filtered, and the liquid was concentrated to dryness to obtain a white solid (S)-N 5 -(2-((1-aminopropyl-2-yl)oxy)-5-fluorobenzyl)-N 5 9.86 g of ethyl-3H-imidazo[4,5-b]pyridine-2,3,5-triamine, yield 88.8%.

[0252] Step 5: (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 Synthesis of amines.

[0253] (S)-N 5 -(2-((1-aminopropyl-2-yl)oxy)-5-fluorobenzyl)-N 5 150 mg (0.40 mmol) of 2,3,5-ethyl-3H-imidazo[4,5-b]pyridine-2,3,5-triamine was dissolved in 5.0 mL of anoxic 1,4-dioxane solution under nitrogen protection, along with 131 mg (0.80 mmol) of triethyl orthoformate. 48 mg (0.80 mmol) of acetic acid was added, and the mixture was heated to 75–80 °C and reacted for 2 hours. HPLC showed that the reaction proceeds were completely reacted. The solvent was removed by concentration under reduced pressure. The residue was dissolved in a suitable amount of dichloromethane, washed twice with NaHCO3, once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Crude product was obtained by column chromatography (DCM:MeOH = 50:1–10:1) to give a white solid (S,Z)-2-ethyl-4-dioxane. 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -Amine 86.0 mg, yield 55.8%. 1H NMR (400MHz, DMSO-d6) δ8.70-8.73 (d, J=12.0Hz, 1H), 7.21-7.36 (m, 2H), 6.86-7 .07(m,4H),6.33-6.39(d,J=8.4Hz,1H),5.76(s,1H),5.41-5.49(d,J=12.4Hz,1 H),4.57-4.62(m,1H),3.84-3.93(m,1H),3.73-3.77(m,1H),3.54-3.63(m,1H), 3.41-3.46(m,2H),3.20-3.27(m,1H),1.27(d,J=6.0Hz,3H),1.16-1.19(m,3H). MS: 384.3 [M+H] + .

[0254] A similar method can be used to prepare compound (A-32): (S,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine: 1 H NMR(400MHz, DMSO-d6)δ8.72(d,J=12.0Hz,1H),7.36~7.28(m,1H),7.21-7.26(m,1H ),7.23-7.25(d,J=8.8Hz,1H),7.07-7.09(d,J=8.8Hz,1H),7.24(d,1H),6.27-6.44( m,1H),6.13(s,1.0H),5.88(s,1H),4.60-4.70(m,1H),3.88-3.93(m,1H),3.50-3.64 (m,2H),3.33`3.42(m,1H),3.21-3.31(m,1H),1.18-1.22(m,3H),1.07-1.14(m,3H). MS: 461.1, 463.1[M+H] + .

[0255] Compound A32 can be prepared by a similar method.

[0256] Example A-33: Preparation of (3S,6S,E)-4 5 -Fluoro-3,6-dimethyl-9-methylthio-1 3H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene((3S,6S,E)-4 5 -Fluoro-3,6-dimethyl-9-methylthio-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzenacyclodecaphan-9-ene):

[0257] Step 1: (3S, 6S, E) - 4 5 -Fluoro-3,6-dimethyl-1 3 Synthesis of H-5-oxo-28,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2-benzodecyclo-9-en-9-thiol):

[0258] 136.8 mg (0.4 mmol) of N5-((S)-1-(2-(((S)-1-aminopropyl-2-yl)oxy)-5-fluorophenyl)-3H-imidazo[4,5-b]pyridine-3,5-diamine was dissolved in 5 mL of anhydrous tetrahydrofuran, and 107.0 mg (0.6 mmol) of thiocarbonyldiimidazole was added. The mixture was stirred at room temperature for 16 h. After the reaction was complete, the solvent was removed by concentration, ethyl acetate was added, and the mixture was washed twice with water. The mixture was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by concentration. The mixture was then chromatographically analyzed using an eluent (DCM:MeOH = 30; 1-20:1) to obtain (3S,6S,E)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-O-28,10-Triaza-1(5,3)-Imidazo[4,5-b]pyridine-4(1,2-benzodecyclo-9-ene-9-thiol product 100 mg, yield 65%).

[0259] Step 2: (3S, 6S, E) - 4 5 -Fluoro-3,6-dimethyl-9-(methylthio)-1 3 Synthesis of -5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyr-4(1,2)-benzodecacyclo-9-ene.

[0260] (3S,6S,E)-4 5 -Fluoro-3,6-dimethyl-1 3H-5-O-28,10-Triaza-1(5,3)-Imidazolo[4,5-b]pyridine-4(1,2-benzodecyclo-9-en-9-thiol 100 mg (0.26 mmol) was dissolved in methanol, and 110 mg (0.78 mmol) of iodomethane was added. The mixture was reacted at 70-75 °C for 3 h. After the reaction was complete, the solvent was removed by concentration, and dichloromethane was added as solvent. The mixture was then directly separated by chromatography using eluent (DCM:MeOH = 20:1) to obtain (3S,6S,E)-4. 5 -Fluoro-3,6-dimethyl-9-(methylthio)-1 3 -5-O-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyrene-4(1,2-benzodecacyclo-9-ene product 42 mg, yield 40.4%, 1 HNMR(400MHz, CDCl3)δ9.16(dd,J=8.6,3.3Hz,1H),7.67(s,1H),7.62(d,J=8.7Hz,1H),6. 92(dd,J=9.9,8.4Hz,1H),6.73(d,J=1.6Hz,1H),6.72(d,J=1.6Hz,1H),6.30(d,J=8.7Hz, 1H),5.40–5.30(m,1H),4.88(d,J=5.3Hz,1H),4.56–4.45(m,1H),3.62–3.48(m,1H),3.27 (ddd,J=14.8,8.9,3.8Hz,1H),2.40(s,3H),1.40(d,J=6.4Hz,3H),1.37(d,J=6.9Hz,3H). MS:401.2[M+H] + .

[0261] Example A-34: (3R,6S,Z)-44-fluoro-3,6-dimethyl-13H-5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]piperidine-4(1,2)-phenylcyclodecanecycloene-9-ene

[0262] Step 1: Synthesis of (S)-(2-(2-acetyl-5-fluorophenoxy)propyl)tert-butyl carbamate

[0263] 4.0 g (25.95 mmol) of 1-(4-fluoro-2-hydroxyphenyl)-1-ethyl ketone, 10.2 g (38.93 mmol) of triphenylphosphine, and 6.8 g (38.93 mol) of (R)-(2-hydroxypropyl)carbamate tert-butyl ester were dissolved in 40 mL of dichloromethane. The mixture was cooled to 0 °C, and 7.2 g (41.52 mol) of diethyl azodicarbonate was slowly added dropwise. After the addition was complete, the reaction was continued for 5 h. TLC showed that the starting material had basically disappeared. 40 mL of petroleum ether was added, and the mixture was stirred to induce crystallization. The solid was filtered, and the filtrate was concentrated. The oily substance was chromatographically analyzed with an eluent (petroleum ether: ethyl acetate = 20:1-15:1) to give 6.48 g of (S)-(2-(2-acetyl-5-fluorophenoxy)propyl)carbamate tert-butyl ester, with a yield of 80.3%.

[0264] Step 2: Synthesis of tert-butyl carbamate ((S)-2-(2-((Z)-1-(((R)-tert-butylsulfinyl)imino)ethyl)-5-fluorophenoxy)propyl)carbamate

[0265] 6.48 g (20.48 mmol) of (S)-(2-(2-acetyl-5-fluorophenoxy)propyl)carbamate tert-butyl ester, 9.51 g (41.68 mmol) of tetraethoxytitanium, and 3.79 g (0.252 mol) of (R)-2-methylpropane-2-sulfinamide were added to 20 mL of anhydrous tetrahydrofuran and 20 mL of anhydrous 2-methyltetrahydrofuran. The mixture was heated to 60-65 °C and reacted under N2 protection for 12 hours. LC showed that the raw material had almost disappeared. The mixture was cooled to room temperature, and water was slowly added dropwise. A solid precipitated out. The solid was filtered and washed once with ethyl acetate. The filtrates and ethyl acetate were combined. The organic phase was washed once with saturated NaHCO3 and twice with saturated NaCl solution. The mixture was concentrated to obtain 8.56 g of crude tert-butyl carbamate ((S)-2-(2-((Z)-1-(((R)-tert-butylsulfinyl)imino)ethyl)-5-fluorophenoxy)propyl)carbamate.

[0266] Step 3: Synthesis of tert-butyl carbamate ((S)-2-(2-((S)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-5-fluorophenoxy)propyl)carbamate

[0267] Dissolve 8.56 g (20.68 mol) of tert-butyl carbamate ((S)-2-(2-((Z)-1-(((R)-tert-butylsulfinyl)imino)ethyl)-5-fluorophenoxy)propyl)carbamate in 90% tetrahydrofuran aqueous solution, cool to -30 to -50 °C, and maintain the temperature below -30 °C. Add 2.41 g (63.49 mmol) of sodium borohydride in portions. After the addition is complete, continue the reaction for 1 hour. Water and ethyl acetate were added, and the mixture was stirred and separated. The organic phase was washed twice with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then subjected to chromatography using petroleum ether:ethyl acetate = 20:1 to 5:1 to obtain 5.40 g of ((S)-2-(2-((S)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-5-fluorophenoxy)propyl)carbamate tert-butyl ester, with a two-step yield of 62.3%.

[0268] Step 4: Synthesis of tert-butyl ((S)-2-(2-((S)-1-aminoethyl)-5-fluorophenoxy)propyl)carbamate

[0269] 5.40 g (12.98 mol) of tert-butyl ((S)-2-(2-((S)-1-(((R)-tert-butylsulfinyl)amino)ethyl)-5-fluorophenoxy)propyl)carbamate was dissolved in 500 mL of 80% tetrahydrofuran aqueous solution. 0.66 g (2.60 mmol) of elemental iodine was added, and the reaction was carried out at room temperature for 5 h. The color of the system changed from brown to pale yellow. TLC showed that the reactants had completely reacted. Saturated sodium thiosulfate solution was added, and the mixture was extracted twice with ethyl acetate. The aqueous phase was then extracted twice more with ethyl acetate. The ethyl acetate phase was washed twice with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain an oily substance. Chromatography using a dichloromethane:methanol eluent (100:1 to 20:1) yielded 2.2 g of the product ((S)-2-(2-((S)-1-aminoethyl)-5-fluorophenoxy)propyl)carbamate, with a yield of 54.3%.

[0270] Step 5: Synthesis of tert-butyl 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-4-fluorophenyl)ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylic acid

[0271] 1.20 g (3.85 mmol) of tert-butyl ((S)-2-(2-((S)-1-aminoethyl)-4-fluorophenoxy)propyl)carbamate was dissolved in 10 mL of dry DMF, followed by the addition of 1.11 g (3.885 mmol) of tert-butyl 2-(6-chloro-3-nitropyridin-2-yl)hydrazide-1-carboxylate and 1.48 g (11.54 mmol) of diisopropylethylamine. The reaction was heated to 80 °C and reacted for 12 h. TLC showed that the reaction was complete. The reaction solution was washed twice with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (PE / EA = 30 / 1~10 / 1, V / V) to give 1.7 g of tert-butyl 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-4-fluorophenyl)ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylate, with a yield of 78.3%.

[0272] Step 6: Synthesis of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butoxycarbonyl)amino)prop-2-yl)oxy)-4-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid

[0273] 1.70 g (48.0 mol) of tert-butyl 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-4-fluorophenyl)ethyl)amino)-3-nitropyridin-2-yl)hydrazine-1-carboxylate (dissolved in oxygen-free THF) was added to a reaction flask under nitrogen protection. 1.70 g of Raney-Ni catalyst was added, followed by the slow addition of 0.48 g (15.07 mmol) of 80% hydrazine hydrate at room temperature. After the addition was complete, the reaction mixture was stirred; the bubbles were colorless, and the system was a transparent liquid. TLC analysis confirmed the reaction was complete. The reaction mixture was filtered quickly through a diatomaceous earth filter under nitrogen atmosphere. The filtrate was concentrated to dryness under reduced pressure to obtain 1.50 g of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-4-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylic acid. The crude product rapidly darkened in color and was used directly in the next reaction without further purification.

[0274] Step 7: Synthesis of tert-butyl carbamate ((2S)-2-(2-(1-((3-((tert-butyloxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-5-fluorophenoxy)propyl)carbamate

[0275] 1.5 g of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-4-fluorophenyl)ethyl)amino)pyridin-2-yl)hydrazine-1-carboxylate was dissolved in 10 mL of oxygen-free 1,4-dioxane and 1.5 g of triethyl orthoformate under nitrogen protection. 0.75 mL of acetic acid was added, and the mixture was heated to 75–80 °C and reacted for 2–3 hours. TLC showed that the reaction proceeded completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of DCM, washed twice with organic phase NaHCO3, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 5:1–3:1) yielded 0.88 g of a white solid (((2S)-2-(2-(1-((3-((tert-butyloxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-5-fluorophenoxy)propyl)carbamate tert-butyl ester). The two-step yield was 71.6%.

[0276] Step 8: Synthesis of N5-((S)-1-(2-(((S)-1-aminopropyl-2-yl)oxy)-4-fluorophenyl)ethyl)-3H-imidazo[4,5-b]pyridine-3,5-diamine

[0277] 0.88 g (1.61 mmol) of tert-butyl carbamate ((2S)-2-(2-(1-((3-((tert-butyloxycarbonyl)amino)-3H-imidazo[4,5-b]pyridin-5-yl)amino)ethyl)-5-fluorophenoxy)propyl)carbamate was dissolved in 2.57 mL of 1,4-dioxane hydrochloric acid solution (1M). The mixture was stirred at room temperature for 1–2 hours, and TLC showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in methanol solution with sodium bicarbonate solid added. The mixture was stirred for 10 minutes, filtered, and the liquid was concentrated to dryness to give 0.53 g of white solid N5-((S)-1-(2-(((S)-1-aminopropyl-2-yl)oxy)-4-fluorophenyl)ethyl)-3H-imidazo[4,5-b]pyridin-3,5-diamine, with a yield of 95.1%.

[0278] Step 9: (3R,6S,Z)-44-fluoro-3,6-dimethyl-13H-5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]piperidine-4(1,2)-phenylcyclodecanecycloene-9-ene

[0279] 130 mg (0.38 mmol) of N5-((S)-1-(2-(((S)-1-aminopropyl-2-yl)oxy)-5-fluorophenyl)ethyl)-3H-imidazo[4,5-b]pyridine-3,5-diamine was dissolved in 5.0 mL of an oxygen-free 1,4-dioxane solution under nitrogen protection, along with 169 mg (1.14 mmol) of triethyl orthoformate. 68.4 mg (1.14 mmol) of acetic acid was added, and the mixture was heated to 75–80 °C and reacted for 2–3 hours. HPLC showed that the reaction proceeded completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of dichloromethane, washed twice with organic phase NaHCO3, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Column chromatography (DCM:MeOH = 50:1–10:1) yielded 86.0 mg of a white solid (3R,6S,Z)-4,4-fluoro-3,6-dimethyl-13H-5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]piperidine-4(1,2)-phenylcyclodecanecycloene-9-ene, with a yield of 63.9%. 1HNMR (400MHz, CDCl3) δ9.40 (d, J = 49.9Hz, 1H), 7.76 (d, J = 24.4Hz, 1H), 7.68 (dd, J = 8 .7,2.8Hz,1H),7.29(dd,J=8.5,6.9Hz,1H),7.23(dd,J=8.4,6.7Hz,1H),6.71–6.46(m ,3H),6.36(dd,J=8.7,5.5Hz,1H),5.65–5.42(m,1H),5.04(t,J=13.5Hz,1H),4.47(d dt,J=13.0,9.9,6.1Hz,1H),3.47(ddd,J=22.6,12.7,11.1Hz,2H),1.53–1.36(m,6H). MS: 355.2[M+H] + .

[0280] The compounds listed below were prepared using a synthetic method similar to that used for A-34:

[0281] Example A-43: (3S,6S,Z)-4 5 -Fluoro-3,6,9-trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzenecyclodecane-9-ene

[0282] N 5-((S)-1-(2-((((S)-1-aminopropane-2-yl)oxy)-5-fluorophenyl)ethyl-3H-pyrido[4,5-b]pyridine-3,5-diamine (100 mg, 0.29 mmol) was dissolved in 5.0 mL of anoxic 1,4-dioxane solution under nitrogen protection. Triethyl orthoacetate (140 mg, 0.87 mmol) and acetic acid (52 mg, 0.87 mmol) were added. The mixture was heated to 75–80 °C and stirred for 2–3 hours. HPLC showed that the starting material reacted completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of dichloromethane, washed twice with NaHCO3 aqueous solution, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude target product. Column chromatography (eluent DCM:MeOH = 50:1–10:1) yielded a white solid (91.0 mg, yield: 83.4%). 1 H NMR(400MHz,DMSO-d6)δ(ppm):8.01(d,1H),7.79(d,1H),7.67(dd,1H),7.12 -7.0(m,3H),6.92(m,1H),6.47(d,J=8.7Hz,1H),5.20(m,1H),4.45(m,1H),4 .25(m,1H),3.47(dd,J=14.9,5.7Hz,1H),3.12–3.14(m,1H),1.85(s,3H),1. 46-1.35(m,2H),1.32(d,J=7.0Hz,3H),1.28(d,J=6.2Hz,3H).MS:368.4[M+H] + .

[0283] Synthesize the following compounds using the same method:

[0284] Example A-46: (3S,6S,Z)-4 5 -Fluoro-9-isopropyl-1 2 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzylcyclodecane-9-ene

[0285] N 5-((S)-1-(2-((((S)-1-aminopropane-2-yl)oxy)-5-fluorophenyl)ethyl)-2-methyl-3H-pyrido[4,5-b]pyridine-3,5-diamine (100 mg, 0.27 mmol) was dissolved in an oxygen-free 1,4-dioxane solution (5.0 mL) under nitrogen protection. Triethyl isobutyrate (170 mg, 0.87 mmol) and acetic acid (52 mg, 0.87 mmol) were then added. The mixture was heated to 75–80 °C and stirred for 2–3 hours. HPLC showed that the reaction proceeds were completely reacted. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of dichloromethane. The organic phase was washed twice with NaHCO3 aqueous solution and once with saturated NaCl solution. It was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH = 50:1 to 10:1) to obtain the target product as a white solid (72.0 mg, yield 65.4%). 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.03 (t, 1H), 7.47 (d, 1H), 7.16 (d, 1H), 7.04 (m, 1H), 6.93 (m, 1H), 6.84 (m, 1H), 6.29 (d, J = 8.7Hz, 1H), 5 .29(m,1H),4.55(m,1H),3.43(m,1H),3.26(m,1H),2.37(s,3H),2.35(m,1H),1.38(d,,3H),1.24(m,3H),0.96(d,6H).MS: 411.4[M+H] + .

[0286] Synthesize the following compounds using the same method:

[0287] Example A-49: (3S,6S,Z)-4 5 -Fluorine-1 7 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzenecyclodecane-9-ene

[0288] Step 1: Synthesis of tert-butyl 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-4-methyl-3-nitropyridin-2-yl)hydrazone-1-carboxylic acid

[0289] ((S)-2-(2-((S)-1-aminoethyl)-4-fluorophenyloxy)propyl tert-butyl carbonate (17.0 g, 54.5 mmol) was dissolved in dry DMF (250 mL), and 2-(6-chloro-4-methyl-3-nitropyridin-2-yl)nitrile-1-carboxylic acid tert-butyl ester (14.90 g, 49.4 mol) and diisopropylethylamine (21.09 g, 163.2 mol) were added at 0 °C. The reaction was heated to 55 °C and reacted for 16 h. TLC showed that the reaction was complete. The reaction solution was slowly poured into an ice-water mixture, and the aqueous layer was extracted once with an ethyl acetate / petroleum ether mixture (400 mL, EA / PE = 1 / 1, v / v). The aqueous phase was discarded. The viscous syrupy substance eluted from the organic layer was washed twice with pure water (150 mL × 2), and then washed once with saturated brine. The organic phase was dried over anhydrous sodium sulfate. Sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (PE / EA = 30 / 1~10 / 1, V / V) to obtain the target product (27.3 g, yield: 93.6%).

[0290] Step 2: Synthesis of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butoxycarbonyl)amino)propyl-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-4-methylpyridin-2-yl)hydrazone-1-carboxylic acid

[0291] 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)propyl-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-4-methyl-3-nitropyridin-2-yl)hydrazone-1-carboxylic acid tert-butyl ester (27.3 g, 48.0 mmol) was dissolved in oxygen-free THF (300 mL). Raney-Ni catalyst (8.2 g, wet weight) was added under nitrogen protection, followed by slow addition of 80% hydrazine hydrate (5.3 g, 132.0 mmol) at room temperature. After the addition was complete, the reaction mixture was stirred until no bubbles were generated and the solution became clear. HPLC analysis confirmed the reaction was complete. The reaction mixture was filtered quickly through a diatomaceous earth filter under nitrogen atmosphere. The filtrate was concentrated to dryness under reduced pressure to obtain the target product (26.7 g) crude product. The crude product rapidly darkened in color and was used directly in the next reaction without further purification.

[0292] Step 3: Synthesis of ((S)-2-(2-((S)-1-((3-((tert-butoxycarbonyl)amino)-7-methyl-3H-pyrido[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenyloxy)propyl)tert-butyl carbonate

[0293] Crude tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)propyl-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-4-methylpyridin-2-yl)hydrazone-1-carboxylic acid (tert-butyl ester) (26.7 g) was dissolved in oxygen-free 1,4-dioxane (100 mL) under nitrogen protection. Triethyl orthoformate (100 mL) and acetic acid (10.0 mL) were then added sequentially. The mixture was heated to 75–80 °C and stirred for 2–3 hours. HPLC showed complete reaction of the starting material. The solvent was removed by vacuum concentration, and the residue was dissolved in an appropriate amount of ethyl acetate. The organic phase was washed twice with NaHCO3 aqueous solution and once with saturated NaCl aqueous solution. The solution was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (PE:EA = 5:1–3:1) to obtain the target product (8.2 g, yield: 69.0%, two steps).

[0294] Step 4: N 5 Synthesis of -((S)-1-(2-(((S)-1-aminopropane-2-yl)oxy)-5-fluorophenyl)ethyl 7-methyl-3H-pyrido[4,5-b]pyridine-3,5-diamine

[0295] ((S)-2-(2-((S)-1-((3-((tert-butoxycarbonyl)amino)-7-methyl-3H-pyrido[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenyloxy)propyl)tert-butyl carbonate (18.2 g, 33.3 mmol) was dissolved in hydrochloric acid-ethanol solution (200 mL, 1 M). The mixture was stirred at room temperature for 1–2 hours, and HPLC showed complete reaction. The reaction solution was concentrated under reduced pressure. Sodium bicarbonate solid was added to the remaining solution in a mixture of dichloromethane and methanol (2:1), and the mixture was stirred for 10 minutes. The mixture was filtered, and the liquid was concentrated to dryness to obtain the crude target product (11.73 g, yield 84.4%).

[0296] Step 5: (3S, 6S) - 4 5 -Fluorine-1 7 3,6-Trimethyl-1 3 Synthesis of H-5-oxa-2,8,10-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzenecyclododecane-9-ene.

[0297] N 5-((S)-1-(2-(((S)-1-aminopropane-2-yl)oxy)-5-fluorophenyl)ethyl 7-methyl-3H-pyrido[4,5-b]pyridine-3,5-diamine (100 mg, 0.29 mmol) was dissolved in anaerobic 1,4-dioxane (5.0 mL) under nitrogen protection, followed by the addition of triethyl orthoformate (128 mg, 0.87 mmol) and acetic acid (52 mg, 0.87 mmol), and the mixture was heated to... The reaction was stirred at 75–80 °C for 2–3 hours, and HPLC showed that the starting material reacted completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of dichloromethane, and the organic phase was washed twice with NaHCO3 aqueous solution, once with saturated NaCl aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (DCM:MeOH = 50:1–10:1) to give a white solid target product (86.0 mg, yield 83.4%). 1 H NMR (400MHz, DMSO-d6) δ (ppm): 8.34 (d, 1H), 7.74 (d, 1H), 7.28 (dd, J = 14.9, 5.7Hz, 1H), 7.20 (d, 1H), 7.05–6. 95(m,3H),6.34(s,1H),5.54(m,1H),4.51(m,1H),3.53(m,2H),3.25(m,1H),2.26(s,3H),1.42–1.27(m,6H). MS: 369.4[M+H] + .

[0298] Synthesize the following compounds using the same method:

[0299] Example A-51: (3S,6S)-4 5 -Fluorine-1 6 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene

[0300] Step 1: Synthesis of tert-butyl 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-)oxy)-5-fluorophenyl)ethyl)amino)-5-methyl-3-nitropyridin-2-yl)hydrazone-1-carboxylic acid

[0301] 17.0 g (54.5 mmol) of ((S)-2-(2-((S)-1-aminoethyl)-4-fluorophenyloxy)propyl tert-butyl carbonate was dissolved in 250 mL of dry DMF. Then, 14.90 g (51.7 mol) of 2-(6-chloro-5-methyl-3-nitropyridin-2-yl)nitrile-1-carboxylic acid tert-butyl ester and 21.0 g (163.0 mol) of diisopropylethylamine were added at 0 °C. The mixture was heated to 55 °C and reacted for 16 h. TLC showed that the reaction was complete. The reaction solution was washed twice with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. The sodium sulfate was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (PE / EA = 30 / 1~10 / 1, V / V) to give 27.3 g of 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-)oxy)-5-fluorophenyl)ethyl)amino)-5-methyl-3-nitropyridin-2-yl)hydrazone-1-carboxylic acid tert-butyl ester, with a yield of 93.6%.

[0302] Step 2: Synthesis of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butoxycarbonyl)amino)-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-5-methylpyridin-2-yl)hydrazone-1-carboxylic acid

[0303] 27.3 g (48.0 mmol) of 2-(6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)prop-2-)oxy)-5-fluorophenyl)ethyl)amino)-5-methyl-3-nitropyridin-2-yl)hydrazone-1-carboxylic acid tert-butyl ester dissolved in oxygen-free THF) was added to a reaction flask under nitrogen protection. 8.2 g of Raney-Ni catalyst was added, followed by the slow addition of 5.3 g (132.0 mmol) of 80% hydrazine hydrate at room temperature. After the addition was complete, the reaction mixture was stirred; the bubbles were colorless, and the system was a transparent liquid. HPLC analysis confirmed the reaction was complete. The reaction mixture was filtered quickly through a diatomaceous earth filter under nitrogen atmosphere. The filtrate was concentrated to dryness under reduced pressure to obtain 26.7 g of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-5-methylpyridin-2-yl)hydrazone-1-carboxylic acid. The crude product rapidly darkened in color and was used directly in the next reaction without further purification.

[0304] Step 3: Synthesis of tert-butyl((S)-2-(2-((S)-1-((3-((tert-butoxycarbonyl)amino)-8-methyl-3H-pyrido[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenyloxy)propyl)carbonic acid

[0305] 26.7 g of tert-butyl 2-(3-amino-6-(((S)-1-(2-((((S)-1-((tert-butyloxycarbonyl)amino)-2-yl)oxy)-5-fluorophenyl)ethyl)amino)-5-methylpyridin-2-yl)hydrazone-1-carboxylic acid was dissolved in 100 mL of oxygen-free 1,4-dioxane and 100 mL of triethyl orthoformate under nitrogen protection. 10.0 mL of acetic acid was added, and the mixture was heated to 75–80 °C and allowed to rest for 2–3 hours. HPLC showed that the reaction proceeded completely. The solvent was removed by concentration under reduced pressure. The residue was dissolved in an appropriate amount of ethyl acetate, washed twice with organic phase NaHCO3, washed once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Column chromatography (PE:EA = 5:1–3:1) yielded 8.2 g of a white solid ((S)-2-(2-((S)-1-((3-((tert-butyloxycarbonyl)amino)-8-methyl-3H-pyrido[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenyloxy)propyl)tert-butyl carbonate. Two-step yield: 69.0%.

[0306] Step 4: N 5 Synthesis of -((S)-1-(2-(((S)-1-aminopropane-2-yl)oxy)-5-fluorophenyl)ethyl 8-methyl-3H-pyrido[4,5-b]pyridine-3,5-diamine

[0307] 18.2 g (33.3 mmol) of ((S)-2-(2-(((S)-1-((3-((tert-butoxycarbonyl)amino)-8-methyl-3H-pyrido[4,5-b]pyridin-5-yl)amino)ethyl)-4-fluorophenyloxy)propyl)tert-butyl carbonate was dissolved in 200 mL of 1 M hydrochloric acid-ethanol solution. The mixture was stirred at room temperature for 1–2 hours, and HPLC showed complete reaction. The reaction solution was concentrated under reduced pressure. Sodium bicarbonate solid was added to the remaining solution in a 2:1 mixture of dichloromethane and methanol, and the mixture was stirred for 10 minutes. The mixture was filtered, and the liquid was concentrated to dryness to give 11.73 g of a white solid, N5-((S)-1-(2-(((S)-1-aminopropane-2-yl)oxy)-5-fluorophenyl)ethyl-8-methyl-3H-pyrido[4,5-b]pyridin-3,5-diamine, with a yield of 84.4%.

[0308] Step 5: (3S, 6S) - 4 5 -Fluorine-1 6 Synthesis of 3,6-trimethyl-13H-5-oxo-2,810-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzocyclodecane-9-ene.

[0309] 100 mg (0.29 mmol) of N5-((S)-1-(2-(((S)-1-aminopropane-2-yl)oxy)-5-fluorophenyl)ethyl 8-methyl-3H-pyrido[4,5-b]pyridine-3,5-diamine was dissolved in 5.0 mL of anoxic 1,4-dioxane solution under nitrogen protection, along with 128 mg (0.87 mmol) of triethyl orthoformate. 52 mg (0.87 mmol) of acetic acid was added, and the mixture was heated to 75–80 °C and reacted for 2–3 hours. HPLC showed complete reaction of the starting material. The solvent was removed by concentration under reduced pressure. The residue was dissolved in a suitable amount of dichloromethane, washed twice with NaHCO3, once with saturated NaCl solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Crude product was obtained by column chromatography (DCM:MeOH = 50:1–10:1) to obtain a white solid (3S,6S,Z)-4 5 -Fluorine-1 6 ,3,6-Trimethyl-13H-5-oxo-2,810-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (86.0 mg, yield 83.4%). 1 H NMR (400MHz, DMSO-d6) δ (ppm): 9.15 (d, 1H), 7.83 (s, 1H), 7.56 (s, 1H), 7.40 (m, 2H),, 7.00–6.92 (m, 2H), 6. 58(d,J=8.7Hz,1H),5.71(m,1H),4.67(m,1H),3.49(m,1H),3.31(m,1H),2.32(s,,3H),1.46–1.35(m,9H). MS: 369.4[M+H] + .

[0310] Biological detection:

[0311] Enzyme activity assay:

[0312] Prepare 2×ATP (25 μM) / substrate solution and 2× kinase solution using an in vitro enzyme activity assay with kinase reaction buffer. Transfer 50 nL of the compound dilution to a 384 assay plate using an Echo 655; centrifuge and add 2.5 μL of 2×Kinase solution to the 384 assay plate, centrifuge at 1000 rpm for 1 min, and incubate at 25 °C for 10 min. Add 2.5 μL of 2× substrate and ATP solution to the 384 assay plate, centrifuge at 1000 rpm for 1 min, and incubate at 25 °C for 40 min. Prepare 2×XL665 and antibody detection reagents with assay buffer. Add 5 μL of kinase detection reagent to the assay plate and incubate at 25 °C. Centrifuge at 1000 rpm for 1 min and incubate at 25 °C for 1 h. Read the fluorescence signals at 620 nm (Cryptate) and 665 nm (XL665) using an EnVision Multilabel Reader (PerkinElmer). % Inhibition = 100% - (Compound Positive Control) / (Negative Control Positive Control) * 100%.

[0313] Enzyme activity assay Note: A: IC 50 ≤10nM; B: 10 <IC 50 ≤100nM; C: 100nM <IC 50 ≤500nM; D: 500nM <IC 50 <1000nM; E: 1.0μM≤IC 50 ≤5.0μM.

[0314] Anti-cell proliferation activity assay

[0315] Cell antiproliferative activity was determined using the CellTiter-Glo (Promega, USA) method. A 1000× compound solution was prepared in DMSO, and a 20× compound solution was prepared by adding 1 μl of the 1000× compound to 49 μl of growth medium. The cell suspension was diluted to the desired density in growth medium, and 95 μl was transferred to a 96-well plate. 5 μl of the 20× compound was added to each well according to the plate plot. The final concentration of dimethyl sulfoxide (DMSO) in each well was 0.1%. Cells were then incubated at 37°C and 5% CO2 for 72 hours, and the plate was equilibrated to room temperature before assay. 20 μl of cell titration-fluorescence reagent was added to each well. The contents were mixed on an orbital vibrator for 2 minutes to induce cell lysis. Cells were incubated at room temperature for 10 minutes to stabilize the luminescence signal. Luminescence was recorded using an EnVision multi-tag reader (PerkinElmer). Cell viability (CV%) relative to the medium (DMSO)-treated control wells was calculated using the following formula: Cell viability (%) = (RLU compound - RLU blank) / (RLU control - RLU blank) * 100%. IC50 values ​​were calculated using GraphPad Prism 6.0 software, and a four-parameter equation was fitted to generate concentration-response curves. All experiments were performed with three parallel samples and three replicates.

[0316] Anti-cell proliferation activity (Ba / F3)

Claims

1. A compound of Formula I, its isomers, prodrugs, solvates, isotopic derivatives, or pharmaceutically acceptable salts. in, R1 is selected from H, R'-S-, R'-O-, and C. 1-6 C substituted with alkyl and halogen or cyano groups 1-6 Alkyl; R' is selected from C 1-6 alkyl; R2 is selected from H and C. 1-6 C substituted with alkyl, halogen or cyano groups 1-6 Alkyl, amino, -NHC 1-6 Alkyl, -N(C) 1-6 alkyl)2-, -SC 1-6 Alkyl, -S(O)-C 1-6 Alkyl, -S(O)2-C 1-6 Alkyl groups, -CONH2, -CONR'R” and C 1-6 Alkoxycarbonyl; wherein "NR'R" is selected from C 1-6 alkyl. R3 and R4 are independently selected from H and C. 1-6 alkyl; R5 and R6 are independently selected from H and C. 1-6 Alkyl groups, or R5 and R6, can be linked to form a five- or six-membered heterocycle, which may optionally be substituted with a halogen or have an oxygen heterocycle atom; R7 can be one to four independently selected from H, halogen, -CN, C 1-6 C substituted with alkyl, halogen or cyano groups 1-6 Alkyl substituents, C 1-6 Alkoxy; X is selected from -CH- and N; L is -CHR 10 -CHR 11 -or-CHR 10 -CH2CHR 11 -, where R 10 and R 11 Independently selected from H and C 1-6 alkyl.

2. The compound of formula I according to claim 1, its isomers, prodrugs, solvates, isotope derivatives, or pharmaceutically acceptable salts. in, R1 is selected from H, R'-S-, C 1-6 Alkyl and F-substituted C 1-6 Alkyl; R' is selected from C 1-6 alkyl; R2 is selected from H and C. 1-6 Alkyl, F-substituted C 1-6 Alkyl, amino and C 1-6 Alkoxycarbonyl; R3 and R4 are independently selected from H and C. 1-6 alkyl; R5 and R6 are independently selected from H and C. 1-6 Alkyl groups, or R5 and R6, can be linked to form a five- or six-membered saturated heterocycle, which may optionally be substituted with F or have -O- heterocyclic atoms; R7 can be one to four independently selected from H, halogen, -CN, C 1-6 Alkyl, halogen-substituted C 1-6 Substituents of alkyl groups; X is selected from -CH- and N; L is -CHR 10 -CHR 11 -or-CHR 10 -CH2CHR 11 -, where R 10 and R 11 Independently selected from H and C 1-6 alkyl.

3. The compound, its isomer, prodrug, solvate, isotope derivative, or pharmaceutically acceptable salt according to claim 1 or 2, wherein, R1 is H.

4. The compound, its isomer, prodrug, solvate, isotope derivative, or pharmaceutically acceptable salt according to any one of claims 1-3, wherein, R2 is selected from H, C1-6 alkyl, amino, and C2. 1-6 Alkoxycarbonyl group.

5. The compound, its isomer, prodrug, solvate, isotope derivative, or pharmaceutically acceptable salt according to any one of claims 1-4, wherein, R3 and R4 are independently selected from H.

6. The compound, its isomer, prodrug, solvate, isotope derivative, or pharmaceutically acceptable salt according to any one of claims 1-5, wherein, R5 and R6 are independently selected from H and C. 1-6 Alkyl groups, or R5 and R6 linked together, form five- or six-membered saturated nitrogen-containing heterocycles.

7. The compound, its isomer, prodrug, solvate, isotope derivative, or pharmaceutically acceptable salt according to any one of claims 1-6, wherein, R1 is selected from H; R2 is selected from H, -NH2, C 1-6 Alkyl and C 1-6 Alkoxycarbonyl; R3 and R4 are independently selected from H; R5 and R6 are independently selected from H and C. 1-6 Alkyl groups, or R5 and R6, can be linked together to form five- or six-membered saturated nitrogen-containing heterocycles; R7 consists of 1-4 independent selections from H, halogens, and -CN; X is selected from -CH- and N; L is -CHR 10 -CHR 11 -、-CHR 10 -CH2CHR 11 -, where R 10 and R 11 Independently selected from H and C 1-6 alkyl.

8. The compound, its isomers, prodrugs, solvates, isotope derivatives, or pharmaceutically acceptable salts according to claim 7, wherein, R7 is selected from H, F, CN, and Br.

9. The compound, its isomer, prodrug, solvate, isotope derivative, or pharmaceutically acceptable salt according to any one of the preceding claims, wherein, R1 is selected from H; R2 is selected from H; Both R3 and R4 are selected from H; R5 is H, R6 is C 1-6 alkyl; R7 is F; X is -CH-; L is selected from -CHCH3-CH2-, -CH2-CHCH3-.

10. The compound, its isomer, prodrug, solvate, isotope derivative, or pharmaceutically acceptable salt according to any one of the preceding claims, wherein, R1 is selected from H; R2 is selected from H; Both R3 and R4 are selected from H; R5 and R6 are connected to form a five- or six-membered saturated nitrogen-containing heterocycle; R7 is F; X is N; L is selected from -CHCH3-CH2-, -CH2-CHCH3-.

11. The compound, its isomers, prodrugs, solvates, isotope derivatives, or pharmaceutically acceptable salts according to claim 1, wherein, The compound is selected from: (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (3S,6S,Z)-4 5 -Fluoro-12,3,6-trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 methyl formate (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (S,Z)-4 6 -bromo-2-ethyl-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S,Z)-2-Ethyl-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (2 2 R,5S,Z)-3 5 -Fluoro-5-methyl-1 3 H-4-oxa-7,9-diaza-1(5,3)-imidazo[4,5-b]pyridine-3(3,2)-pyridine-2(1,2)-pyrrolidinecyclonon-8-ene (S,Z)-2-ethyl-4 5 -Fluoro-12,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (R,Z)-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,9,11-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocycloundecane-10-ene (R,Z)-4 6 -bromo-2-ethyl-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S,Z)-4 6 -bromo-2-ethyl-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (R,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (R,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (R,Z)-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (S,Z)-2-ethyl-4 5 -Fluoro-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (S,Z)-2-Ethyl-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (R,Z)-2-ethyl-7-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (2 2 R,6R,Z)-3 5 -Fluoro-6-methyl-1 3 H-4-oxa-7,9-diaza-1(5,3)-imidazo[4,5-b]pyridine-3(3,2)-pyridine-2(1,2)-pyrrolidinecyclonon-8-ene (2 2 R,6S,Z)-3 5 -Fluoro-6-methyl-1 3 H-4-oxa-7,9-diaza-1(5,3)-imidazo[4,5-b]pyridine-3(3,2)-pyridine-2(1,2)-pyrrolidinecyclonon-8-ene (R,Z)-2-ethyl-4 5 -Fluorine-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (S,Z)-2-ethyl-4 5 -Fluorine-1 2 ,6-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (S,Z)-2-ethyl-4 5 -Fluorine-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-4 6 -formonitrile (S,Z)-4 6 -bromo-2-ethyl-1 2 ,6-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (R,Z)-4 6 -bromo-2-ethyl-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S,Z)-4 6 -bromo-2-ethyl-1 2 ,7-Dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (S,Z)-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine (S,Z)-4 6 -bromo-2-ethyl-4 5 -Fluoro-6-methyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene-1 2 -amine (3S,6S,E)-4 5 -Fluoro-3,6-dimethyl-9-methylthio-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene (3R,6S,Z)-44-fluoro-3,6-dimethyl-13H-5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]piperidine-4(1,2)-phenylcyclodecane-9-ene (3R,6S,Z)-3,6-dimethyl-13H-5-oxo-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene 5 -formonitrile (3R,6S,Z)-3,6-dimethyl-45-(trifluoromethyl)-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzonium-9-ene (3R,6S,Z)-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2-benzocyclodecane-9-ene (R,Z)-45-fluoro-3-methyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2-benzocyclodecane-9-ene (3R,6S,Z)-45-chloro-3,6-dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzocyclodecane-9-ene (3R, 6S, Z)-43,4-5-difluoro-3,6-dimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2-benzocyclodecane-9-ene ((3R, 6S, Z)-45,3,6-trimethyl-13H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4-(1,2)-benzocyclodecane-9-ene (3R, 6S, Z)-4 5 -Methoxy-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-benzocyclodecane-9-ene. (3S,6S,Z)-4 5 -Fluoro-3,6,9-trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzenecyclodecane-9-ene (3S,6S,Z)-4 5 -Fluoro-9-isopropyl-3,6-dimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene (3S,6S,Z)-4 5 -Fluoro-3,6-dimethyl-9-propyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene (3S,6S,Z)-4 5 -Fluoro-9-isopropyl-1 2 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzylcyclodecane-9-ene (3S,6S,Z)-4 5 -Fluorine-1 2 3,6,9-Tetramethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-pyrido[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene (3S,6S,Z)-4 5 -Fluorine-1 2 3,6-Trimethyl-9-propyl-1 3 H-5-O-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene (3S,6S,Z)-4 5 -Fluorine-1 7 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazolium[4,5-b]pyridine-4(1,2)benzenecyclodecane-9-ene (3S,6S,Z)-4 5 -Fluorine-1 7 3,6,9-Tetramethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)-phenylcyclodecane-9-ene (3S,6S)-4 5 -Fluorine-1 6 3,6-Trimethyl-1 3 H-5-oxa-2,8,10-triaza-1(5,3)-imidazo[4,5-b]pyridine-4(1,2)benzenecyclodecane-9-ene.

12. A pharmaceutical composition comprising the compound of any one of claims 1 to 11, an isomer thereof, a prodrug, a solvate, an isotope derivative or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

13. Use of the compound, its isomer, prodrug, solvate, isotope derivative or pharmaceutically acceptable salt of any one of claims 1 to 11 in the preparation of a medicament for inhibiting TRK, ROS1, c-Met, CSF1 and / or ALK.

14. The use according to claim 13, wherein, The drug is used to prevent and treat diseases mediated by TRK, ROS1, c-Met, CSF1 and / or ALK, including pain, cancer and ubiquitination.

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