Nitrogen-containing fused ring compounds, processes for their preparation and medical uses thereof

Abstract

Disclosed are nitrogen-containing fused ring compounds, and preparation methods and medical uses thereof. Specifically disclosed are nitrogen-containing fused ring compounds represented by general formula (I), preparation methods thereof, pharmaceutical compositions containing the same, and uses thereof as RET kinase inhibitors for treating diseases related to RET kinase activity. Definitions of the substituents in general formula (I) are the same as those in the specification.

Description

Technical Field

[0001] This invention relates to nitrogen-containing fused-ring compounds, their preparation methods, and pharmaceutical uses. Specifically, this invention relates to nitrogen-containing fused-ring compounds of general formula (I), their preparation methods, pharmaceutical compositions containing them, and their use as RET kinase inhibitors for the treatment of diseases related to RET kinase activity. Background Technology

[0002] The RET gene, a transfection rearrangement gene, is a proto-oncogene encoding a receptor tyrosine kinase (RTK) that is crucial for many physiological functions, such as early embryogenesis, intestinal development, the nervous system, kidney morphogenesis, spermatogenesis, hematopoiesis, and potential immune regulation. Variations in the RET gene lead to numerous pathologies. On one hand, loss-of-function mutations in RET are a known genetic cause of congenital megacolon, characterized by chronic constipation leading to intestinal obstruction, vomiting, and an increased risk of enterocolitis. On the other hand, aberrant RET kinase receptor activation caused by acquired functional rearrangements and mutations is associated with many tumors. In the past, this gene was primarily considered for the early diagnosis of breast cancer and hereditary medullary thyroid carcinoma (MTC). However, increasing evidence in recent years suggests that aberrant RET activation is a key driver of tumor growth and proliferation (AACR, 2020, 26, 6102-6111).

[0003] RET gene fusions occur in approximately 1%–2% of patients with non-small cell lung cancer (NSCLC) and 10%–20% of patients with papillary thyroid carcinoma (which accounts for about 85% of all thyroid cancers). RET gene mutations occur in approximately 60% of patients with medullary thyroid carcinoma.

[0004] Currently, the main treatment for RET fusion-mutant tumors is the use of multi-kinase inhibitors, such as cabozantinib and vandetanib. However, due to their low selectivity, their efficacy is very limited and their toxicity is significant. Novel selective RET kinase inhibitors, selpercatinib and pralsetinib, may address the off-target toxicity issue. These two highly selective RET kinase inhibitors, having demonstrated good antitumor activity and safety in advanced RET fusion-positive NSCLC, were approved by the US FDA on May 8, 2020, and September 4, 2020, respectively. Despite the encouraging efficacy of highly selective RET tyrosine kinase inhibitors (RET-TKIs), experience with targeted therapy for NSCLC indicates that drug resistance remains an unavoidable and significant problem when using RET-TKIs to treat RET fusion-positive NSCLC. Acquired resistance will eventually limit the duration of the benefits of highly selective RET kinase inhibitors. Therefore, understanding the resistance mechanisms of RET kinase inhibitors and providing novel treatment strategies to overcome resistance is crucial.

[0005] Studies have shown that RET mutations involving the solvent-front residue G810 are one of the resistance mechanisms to highly selective RET kinase inhibitors, while acquired double mutations of both the solvent-front G810 and gatekeeper V804 also account for a certain proportion (Journal of Thoracic Oncology, 2020). Additionally, some resistance is driven by RET-independent resistance (e.g., MET amplification) (Ann Oncol, 2020).

[0006] Therefore, it is necessary to develop a new generation of RET kinase inhibitors or combination therapy strategies that cover more comprehensive resistance sites in order to effectively overcome drug resistance in these patients. Summary of the Invention

[0007] Through dedicated research, the inventors have designed and synthesized a series of nitrogen-containing fused-ring compounds that exhibit inhibitory activity against RET kinases and can be developed into drugs for the prevention or treatment of diseases related to RET kinase activity.

[0008] Therefore, the object of the present invention is to provide a compound of general formula (I) or a meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0009]

[0010] Ring A is selected from cycloalkyl, heterocyclic, aryl, and heteroaryl groups, wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further modified by one or more R groups. 5 Replaced;

[0011] Cycloyl group B is selected from cycloalkyl, heterocyclic, aryl, and heteroaryl groups, wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further modified by one or more R groups. 6 Replaced;

[0012] R 1 The group is selected from hydrogen, deuterium, alkyl, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0013] R 2 and R 3 Each is independently selected from hydrogen, deuterium, alkyl, deuterated alkyl, and haloalkyl;

[0014] Or, R 2 and R3 The C atoms connected to them together form C=O or cycloalkyl groups, which may optionally be further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, and hydroxyalkyl.

[0015] R 4 The group is selected from hydrogen, deuterium, alkyl, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0016] R 5 Selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, nitro, cyano, oxo, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, and -(CH2). q R a -(CH2) q OR a -(CH2) q C(O)R a -(CH2) q C(O)OR a -(CH2) q OC(O)R a -(CH2) q C(O)NR b R c -(CH2) q S(O) p R a -(CH2) q NR b R c -(CH2) q S(O) p NR b R c -NR a C(O)NR b R c -(CH2) q NR b C(O)R a -(CH2) q NR b C(O)OR a Or -(CH2) q NR b S(O)p R a The alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups may optionally be further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0017] Or, two adjacent R 5 Together with the atoms attached thereto, they form cycloalkyl, heterocyclic, aryl, and heteroaryl groups, wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0018] R 6 Selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, nitro, cyano, oxo, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, heteroaryl, and -(CH2). q R a -(CH2) q OR a -(CH2) q C(O)R a -(CH2) q C(O)OR a -(CH2) q OC(O)R a -(CH2) q C(O)NR b R c -(CH2) q S(O) p R a -(CH2) q NR b R c -(CH2) q S(O) p NR b R c -NR a C(O)NR b R c -(CH2) q NR b C(O)R a -(CH2) q NR b C(O)ORa Or -(CH2) q NR b S(O) p R a The alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups may optionally be further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0019] Or, two adjacent R 6 Together with the atoms attached thereto, they form cycloalkyl, heterocyclic, aryl, and heteroaryl groups, wherein the cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0020] R a The group is selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, mercapto, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, alkylsulfonyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0021] R b and R c Each of the following groups is independently selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, mercapto, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups are optionally further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, alkylsulfonyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups;

[0022] Or, R b Or R cTogether with the nitrogen atom attached thereto, a heterocyclic group is formed, wherein the heterocyclic group is optionally further substituted by one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxy, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, alkylsulfonyl, cycloalkyl, heterocyclic, aryl, and heteroaryl;

[0023] p is 0, 1, or 2;

[0024] q is an integer from 0 to 6.

[0025] In one specific embodiment, the compound of general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, is a compound of general formula (II), or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0026]

[0027] Among them, rings A, B, X, Z, and R 1 As defined by general formula (I).

[0028] In another specific embodiment, the compound of general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0029] X is selected from CR 2 R 3 ;

[0030] R 2 R 3 As defined in general formula (I).

[0031] In another specific embodiment, the compound represented by general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: ring A is selected from 5 to 10-membered heteroaryl groups, preferably 5 to 6-membered heteroaryl groups, more preferably pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, pyridinyl, and pyrimidinyl groups; optionally further divided by one or more R groups. 5 Replaced; R 5 As defined by general formula (I).

[0032] In another specific embodiment, the compound represented by general formula (I) according to the present invention, or its meso compound, racemic mixture, enantiomer, diastereomer, mixture thereof, or pharmaceutically acceptable salt thereof, wherein: ring B is selected from 5 to 10-membered heteroaryl or 4 to 6-membered heterocyclic groups, preferably 5 to 6-membered heteroaryl or 5 to 6-membered heterocyclic groups, more preferably pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thiazolyl, pyridinyl, tetrahydropyranyl, dihydropyranyl, thiophenyl, furanyl, tetrahydrofuranyl, oxazolyl, and isoxazolyl; optionally further divided by one or more R 6 Replaced; R 6 As defined by general formula (I).

[0033] In another specific embodiment, the compound represented by general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: ring B is selected from pyridinyl, pyrimidinyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, thiazolyl, pyridinyl, tetrahydropyranyl, dihydropyranyl, thiophenyl, furanyl, tetrahydrofuranyl, oxazolyl, and isoxazolyl; optionally further divided by one or more R... 6 Replaced; R 6 As defined by general formula (I).

[0034] In another specific embodiment, the compound of general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, is a compound of general formula (III), or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0035]

[0036] in,

[0037] Z is selected from C or N;

[0038] Ring B is selected from 5- to 10-membered heteroaryl groups or 4- to 6-membered heterocyclic groups, preferably 5- to 6-membered heteroaryl and 5- to 6-membered heterocyclic groups, more preferably pyrazolyl, imidazolyl, triazolyl, tetrazolyl, thiazolyl, pyridinyl, tetrahydropyranyl, dihydropyranyl, thiophenyl, furanyl, tetrahydrofuranyl, oxazolyl, and isoxazolyl, optionally further surrounded by one or more R groups. 6 Replaced;

[0039] R 1 R 2 R 3 R 5 R 6 As defined by general formula (I).

[0040] In another specific embodiment, the compound of general formula (III) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from pyridinyl, pyrimidinyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, thiazolyl, pyridinyl, tetrahydropyranyl, dihydropyranyl, thiophenyl, furanyl, tetrahydrofuranyl, oxazolyl, and isoxazolyl, optionally further radicalized by one or more R... 6 Replaced; R 6 As defined by general formula (I).

[0041] In another specific embodiment, the compound of general formula (III) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from pyridinyl, pyrazolyl, imidazoleyl, triazolyl, and tetrazolyl; optionally further influenced by one or more R... 6 Replaced; R 6 The group is selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl, wherein the C1-C6 alkyl and C3-C6 cycloalkyl are optionally further substituted by one or more substituents selected from halogen, cyano, hydroxy, C1-C6 alkyl, and C1-C6 alkoxy.

[0042] In another specific embodiment, the compound represented by general formula (I) according to the present invention, or a meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein Z is N.

[0043] In another specific embodiment, the compound of general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, is a compound of general formula (IV), or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0044]

[0045] in,

[0046] X1 is CH or N;

[0047] X2 is CH or N;

[0048] R 1 R 2 R 3 R 5 R 6 As defined by general formula (I).

[0049] In another specific embodiment, the compound represented by general formula (I) according to the present invention, or a meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein Z is C.

[0050] In another specific embodiment, the compound represented by general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, is a compound represented by general formula (V), or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0051]

[0052] Wherein, X3 is CH2, O, S or NH;

[0053] R 1 R 2 R 3 R 5 R 6 As defined by general formula (I).

[0054] In another specific embodiment, the compound of general formula (I) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, is a compound of general formula (VI), or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0055]

[0056] Among them, X5, X6, and X7 are each independently selected from CH or N;

[0057] R 1 R 2 R 3 R 5 R 6 As defined by general formula (I).

[0058] In another specific embodiment, the compound represented by general formulas (I) to (VI) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R 2 and R 3 Each is independently selected from hydrogen, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl, and C1-C6 haloalkyl; preferably, R 2 and R 3Each is independently selected from hydrogen and C1-C6 alkyl; more preferably, R 2 and R 3 It is hydrogen.

[0059] In another specific embodiment, the compound represented by general formulas (I) to (VI) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R 5 The group is selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 deuterated alkoxy, C1-C6 haloalkoxy, C3-C6 cycloalkyl, and 5-6 membered heterocyclic groups; preferably, hydrogen, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 haloalkyl, and C3-C6 cycloalkyl; more preferably, C3-C6 cycloalkyl.

[0060] In another specific embodiment, the compound represented by general formulas (I) to (VI) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R 6 The group is selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and 5- to 6-membered heterocyclic groups, wherein the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, and 5- to 6-membered heterocyclic groups are optionally further substituted by one or more substituents selected from halogen, amino, cyano, and hydroxyl groups; preferably, R 6 The group is selected from hydrogen, deuterium, amino, and C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally further substituted with one or more substituents selected from halogen, amino, cyano, and hydroxyl groups. In another specific embodiment, the compound represented by general formulas (I) to (VI) according to the present invention, or its meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, wherein R 6 It is selected from hydrogen, amino, C1-C6 alkyl, and C1-C6 cyanoalkyl.

[0061] In another specific embodiment, the compound represented by general formulas (I) to (VI) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R 6 The group is selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl, wherein the C1-C6 alkyl and C3-C6 cycloalkyl are optionally further substituted by one or more substituents selected from halogen, cyano, hydroxy, C1-C6 alkyl, and C1-C6 alkoxy.

[0062] In another specific embodiment, the compound represented by general formulas (I) to (VI) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 Selected from hydrogen, deuterium, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, 3- to 8-membered heterocyclic groups, C6-C 10 aryl or 5- to 10-membered heteroaryl, wherein the C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 haloalkyl, C3-C8 cycloalkyl, 3- to 8-membered heterocyclic group, C6-C 10 aryl or 5- to 10-membered heteroaryl, optionally further selected from deuterium, halogen, amino, oxo, nitro, cyano, hydroxyl, mercapto, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, 3- to 6-membered heterocyclic, C6-C 10 The aryl group is substituted with one or more substituents of a 5- to 10-membered heteroaryl group; R 1 Preferably, it is derived from hydrogen, C1-C6 alkyl, C1-C6 deuterated alkyl, C1-C6 haloalkyl, or C3-C6 cycloalkyl, wherein the C3-C6 cycloalkyl may optionally be further substituted with C1-C6 alkyl.

[0063] In another specific embodiment, the compound represented by general formulas (I) to (VI) according to the present invention, or a meso compound, racemic mixture, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, wherein R 1 It is selected from hydrogen, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, and C1-C6 alkyl-C3-C6 cycloalkyl.

[0064] Typical compounds of this invention include, but are not limited to:

[0065]

[0066]

[0067]

[0068]

[0069] It may be in the form of its meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof.

[0070] The present invention further relates to a method for preparing a compound of general formula (IV) or a meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, comprising the following steps:

[0071]

[0072] Compounds of formula IVj undergo cyclization in the presence of a catalyst to give compounds of general formula (IV); the catalyst is preferably glacial acetic acid;

[0073] Among them, X1, X2, R 1 R 2 R 3 R 5 R 6 As defined by general formula (IV).

[0074] The present invention further provides a pharmaceutical composition comprising a compound of formula (I) to formula (VI) according to the present invention, or a meso compound, racemic compound, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0075] The present invention further relates to the use of compounds of general formulas (I) to (VI) according to the present invention, or their meso, racemic, enantiomers, diastereomers, or mixtures thereof, or their pharmaceutically acceptable salts or pharmaceutical compositions comprising thereof, in the preparation of RET kinase inhibitors.

[0076] The present invention further relates to the use of compounds of general formulas (I) to (VI) according to the present invention, or their meso, racemic, enantiomers, diastereomers, or mixtures thereof, or their pharmaceutically acceptable salts or pharmaceutical compositions comprising thereof, in the preparation of medicaments for the prevention and / or treatment of diseases associated with RET kinase activity, preferably malignant neoplasms such as non-small cell lung cancer or thyroid cancer.

[0077] The present invention further relates to a method for inhibiting RET kinase, comprising administering to a patient in need an effective amount of a compound of formula (I) to formula (VI) according to the present invention, or a racemic, racemic, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising thereof.

[0078] The present invention further relates to a method for preventing and / or treating diseases associated with RET kinase activity, comprising administering to a patient in need an effective amount of a compound of formula (I) to formula (VI) according to the present invention, or a racemic, racemic, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising thereof, wherein the disease is preferably a malignant tumor disease, such as non-small cell lung cancer or thyroid cancer.

[0079] The present invention further relates to compounds of general formulas (I) to (VI) according to the present invention, or to racemic, racemic, enantiomer, diastereomer, or mixture thereof, or to pharmaceutically acceptable salt thereof, or to pharmaceutical compositions comprising thereof, for use as RET kinase inhibitors.

[0080] The present invention further relates to compounds of general formulas (I) to (VI) according to the present invention, or their meso, racemic, enantiomers, diastereomers, or mixtures thereof, or their pharmaceutically acceptable salts or pharmaceutical compositions comprising thereof, for the prevention and / or treatment of diseases associated with RET kinase activity, wherein the diseases are preferably malignant tumor diseases, such as non-small cell lung cancer or thyroid cancer.

[0081] According to conventional methods in the field to which this invention pertains, the compounds of this invention can react with acids to form pharmaceutically acceptable acid addition salts. The acids include inorganic and organic acids, with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, etc., being particularly preferred.

[0082] According to conventional methods in the field of this invention, the compounds of this invention can react with a base to form a pharmaceutically acceptable basic addition salt. The base includes inorganic and organic bases; acceptable organic bases include diethanolamine, ethanolamine, N-methylglucosamine, triethanolamine, tromethamine, etc., and acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, and sodium hydroxide, etc.

[0083] Pharmaceutical compositions containing an active ingredient can be in forms suitable for oral administration, such as tablets, sugar lozenges, tablets, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions can be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from sweeteners, flavoring agents, coloring agents, and preservatives to provide an appealing and palatable pharmaceutical formulation. Tablets contain an active ingredient and non-toxic, pharmaceutically acceptable excipients suitable for tablet preparation for mixing. These excipients can be inert excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating agents and disintegrants such as microcrystalline cellulose, croscarmellose sodium, corn starch, or alginate; binders such as starch, gelatin, polyvinylpyrrolidone, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. These tablets may be uncoated or coated using known techniques that provide sustained release over a longer period of time by masking the taste of the drug or by delaying disintegration and absorption in the gastrointestinal tract. For example, water-soluble taste-masking substances such as hydroxypropyl methylcellulose or hydroxypropyl cellulose may be used, or time-extending substances such as ethylcellulose or cellulose acetate butyrate may be used.

[0084] Oral formulations can also be provided in hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or in soft gelatin capsules in which the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oil solvent such as peanut oil, liquid paraffin or olive oil.

[0085] Aqueous suspensions contain active substances and excipients suitable for preparing aqueous suspensions for mixing. Such excipients are suspending agents, such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinylpyrrolidone, and gum arabic; dispersants or wetting agents, which may be naturally occurring phospholipids such as lecithin, or condensation products of olefinic oxygen and fatty acids, such as polyoxyethylene stearate, or condensation products of ethylene oxide and long-chain fatty alcohols, such as heptadecaethyleneoxy cetanol, or condensation products of ethylene oxide and partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitan monooleate, or condensation products of ethylene oxide and partial esters derived from fatty acids and hexitol anhydrides, such as polyoxyethylene dehydrated sorbitan monooleate. Aqueous suspensions may also contain one or more preservatives such as ethylparaben or n-propylparaben, one or more colorants, one or more flavoring agents, and one or more sweeteners such as sucrose, saccharin, or aspartame.

[0086] Oil suspensions can be formulated by suspending the active ingredient in vegetable oils such as peanut oil, olive oil, sesame oil, or coconut oil, or mineral oils such as liquid paraffin. Oil suspensions may contain thickeners such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavoring agents mentioned above can be added to provide a palatable formulation. These compositions can be preserved by adding antioxidants such as butylated hydroxyanisole (BHA) or α-tocopherol.

[0087] The pharmaceutical compositions of the present invention may also be in the form of an oil-in-water emulsion. The oil phase may be a vegetable oil, such as olive oil or peanut oil, or a mineral oil, such as liquid paraffin, or a mixture thereof. Suitable emulsifiers may be naturally occurring phospholipids, such as soybean lecithin, and esters or metaesters derived from fatty acids and hexitan anhydrides, such as sorbitan monooleate, and condensation products of said metaesters and ethylene oxide, such as poly(ethylene oxide) sorbitan monooleate. The emulsion may also contain sweeteners, flavoring agents, preservatives, and antioxidants. Syrups and elixirs formulated with sweeteners such as glycerin, propylene glycol, sorbitol, or sucrose may be used. Such formulations may also contain moderating agents, preservatives, coloring agents, and antioxidants.

[0088] The pharmaceutical compositions of the present invention can be in the form of sterile injectable aqueous solutions. Acceptable solvents and media that can be used include water, Ringer's solution, and isotonic sodium chloride solution. The sterile injectable formulation can be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then treated with a mixture of water and glycerol to form a microemulsion. The injection solution or microemulsion can be injected into the patient's bloodstream by local large-volume injection. Alternatively, it is preferable to administer the solution and microemulsion in a manner that maintains a constant circulating concentration of the compounds of the present invention. To maintain such a constant concentration, a continuous intravenous delivery device can be used.

[0089] The pharmaceutical compositions of the present invention can be in the form of sterile injectable aqueous or oil suspensions for intramuscular and subcutaneous administration. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents as described above, according to known techniques. The sterile injectable formulations can also be sterile injectable solutions or suspensions prepared in non-toxic, parenteral-acceptable diluents or solvents, such as solutions prepared in 1,3-butanediol. Furthermore, sterile fixative oils can be conveniently used as solvents or suspension media. For this purpose, any blended fixative oil, including synthetic mono- or diglycerides of glycerol, can be used. Additionally, fatty acids such as oleic acid can also be used to prepare injectable formulations.

[0090] The compounds of the present invention can be administered in suppository form for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable, non-irritating excipient that is solid at normal temperatures but liquid in the rectum, and thus dissolves in the rectum to release the drug. Such substances include cocoa butter, glycerin gelatin, hydrogenated vegetable oils, polyethylene glycol of various molecular weights, and mixtures of fatty acid esters of polyethylene glycol.

[0091] Those skilled in the art will recognize that the dosage of a drug depends on a variety of factors, including, but not limited to, the activity of the specific compound used, the patient's age, weight, health condition, lifestyle, diet, timing of administration, route of administration, rate of excretion, and combination of drugs. Furthermore, optimal treatment modalities, such as treatment patterns, daily dosage of general formula compounds, or types of pharmaceutically acceptable salts, can be validated based on conventional treatment protocols.

[0092] This invention can contain a compound of a general formula, and its pharmaceutically acceptable salts, hydrates, or solvates as active ingredients, mixed with a pharmaceutically acceptable carrier or excipient to prepare a composition, and formulated into a clinically acceptable dosage form. Derivatives of this invention can be used in combination with other active ingredients, provided they do not produce other adverse effects, such as allergic reactions. The compounds of this invention can be used as the sole active ingredient or in combination with other drugs for treating diseases related to RET kinase activity. Combination therapy is achieved by administering the various therapeutic components simultaneously, separately, or sequentially.

[0093] Detailed description of the invention

[0094] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0095] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight-chain or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, and more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-Dimethylpentyl, 2,2-Dimethylpentyl, 3,3-Dimethylpentyl, 2-Ethylpentyl, 3-Ethylpentyl, n-Octyl, 2,3-Dimethylhexyl, 2,4-Dimethylhexyl, 2,5-Dimethylhexyl, 2,2-Dimethylhexyl, 3,3-Dimethylhexyl, 4,4-Dimethylhexyl, 2-Ethylhexyl, 3-Ethylhexyl, 4-Ethylhexyl, 2-Methyl-2-Ethylpentyl, 2-Methyl-3-Ethylpentyl, n-Nonyl, 2-Methyl-2-Ethylhexyl, 2-Methyl-3-Ethylhexyl, 2,2-Diethylpentyl, n-Decyl, 3,3-Diethylhexyl, 2,2-Diethylhexyl, and their various branched isomers, etc. More preferably, lower alkyl groups containing 1 to 6 carbon atoms are used. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group can be substituted or unsubstituted. When substituted, the substituent can be substituted at any usable connection point. The substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group.

[0096] The term "alkenyl" refers to an alkyl group as defined above, consisting of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2-, or 3-butenyl, etc. Alkenyl groups can be substituted or unsubstituted; when substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0097] The term "alkynyl" refers to an alkyl group as defined above, consisting of at least two carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, propynyl, butynyl, etc. The alkynyl group can be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, and heterocycloalkylthio.

[0098] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclohepttrienyl, cyclooctyl, etc.; polycyclic cycloalkyl groups include spirocyclic, fused-ring, and bridged-ring cycloalkyl groups.

[0099] The term "spirocycloalkyl" refers to a polycyclic group consisting of 5 to 20 quintile rings sharing a single carbon atom (called a spiro atom), which may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Preferably, it is 6 to 14 quintiles, more preferably 7 to 10 quintiles. Spirocycloalkyl groups are classified into monospirocycloalkyl, bispirocycloalkyl, or polyspirocycloalkyl groups based on the number of shared spiro atoms between the rings, with monospirocycloalkyl and bispirocycloalkyl groups being preferred. More preferably, it is a 4-quintile, 4-quintile, 4-quintile, 5-quintile, or 5-quintile / 6-quintile monospirocycloalkyl group. Non-limiting examples of spirocycloalkyl groups include:

[0100]

[0101] The term "fused-ring alkyl" refers to a 5- to 20-membered polycyclic carbon group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a fully conjugated π-electron system. Preferably, it is 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of constituent rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused-ring alkyl, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of fused-ring alkyl include:

[0102]

[0103] The term "bridged cycloalkyl" refers to a 5- to 20-membered polycyclic carbon group in which any two rings share two non-directly bonded carbon atoms. It may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Preferably, it is 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl groups include:

[0104]

[0105] The cycloalkyl ring may be fused to an aryl, heteroaryl, or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, and non-limiting examples include indanyl, tetrahydronaphthyl, benzocycloheptyl, etc. The cycloalkyl group may be optionally substituted or unsubstituted; when substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group.

[0106] The term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which are selected from nitrogen, oxygen, or S(O). m(Where m is an integer from 0 to 2) heteroatoms, but excluding the ring portions of -OO-, -OS-, or -SS-, with the remaining ring atoms being carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; most preferably, it contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably, it contains 5 to 7 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolyl, tetrahydrofuranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably 1, 2, 5-oxadiazolyl, pyranyl, or morpholinyl. Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups.

[0107] The term "spiroheterocyclic group" refers to a polycyclic heterocyclic group consisting of 5 to 20 member monocyclic rings sharing a single atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O). m The ring atoms are (where m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a fully conjugated π-electron system. Preferably, it is 6 to 14-membered, more preferably 7 to 10-membered. Spirocyclic groups are classified into monospirocyclic, bispirocyclic, or multispirocyclic groups according to the number of shared spiroatoms between rings, preferably monospirocyclic and bispirocyclic groups. More preferably, it is a 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospirocyclic group. Non-limiting examples of spirocyclic groups include:

[0108]

[0109] The term "fused heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with the other rings in the system. One or more rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. One or more ring atoms are selected from nitrogen, oxygen, or S(O). m (where m is an integer from 0 to 2) heteroatoms, with the remaining ring atoms being carbon. Preferably, it consists of 6 to 14 rings, more preferably 7 to 10 rings. Depending on the number of rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic group, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

[0110]

[0111] The term "bridged heterocyclic group" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two non-directly bonded atoms. It may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. One or more ring atoms are selected from nitrogen, oxygen, or S(O).m (where m is an integer from 0 to 2) heteroatoms, with the remaining ring atoms being carbon. Preferably, it consists of 6 to 14 rings, more preferably 7 to 10 rings. Depending on the number of rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic group, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

[0112]

[0113] The heterocyclic ring may be fused to an aryl, heteroaryl, or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, and non-limiting examples include:

[0114] wait.

[0115] The heterocyclic group can be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl, or carboxylic acid ester group.

[0116] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a conjugated π-electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. Phenyl is more preferred. The aryl ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, and non-limiting examples include:

[0117]

[0118] The aryl group can be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylic acid ester group.

[0119] The term "heteroaryl" refers to a heteroaryl system comprising 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5 to 10-membered, containing 1 to 3 heteroatoms; more preferably 5- or 6-membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furanyl, thiophene, thiazolyl, pyrazolyl, oxazolyl, pyrroleyl, tetrazolyl, pyridinyl, pyrimidinyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, thiazolyl, pyrazolyl or pyrimidinyl, thiazolyl; more preferably pyrazolyl or thiazolyl. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0120]

[0121] The heteroaryl group can be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylic acid ester group.

[0122] The term "alkoxy" refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), wherein alkyl and cycloalkyl are defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexoxy. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl, or carboxylic acid ester group.

[0123] The term "halogenated alkyl" refers to an alkyl group that has been substituted with one or more halogens, wherein the alkyl group is as defined above.

[0124] The term "haloalkoxy" refers to an alkoxy group that is substituted with one or more halogens, wherein the alkoxy group is as defined above.

[0125] The term “deuterated alkyl” refers to an alkyl group that is substituted with one or more deuterium atoms, wherein the alkyl group is as defined above.

[0126] The term “deuterated alkoxy” refers to an alkyl group substituted with one or more deuterium atoms, wherein the alkoxy group is as defined above.

[0127] The term "hydroxyalkyl" refers to an alkyl group that is substituted with one or more hydroxyl groups, wherein the alkyl group is as defined above.

[0128] The term "hydroxyl group" refers to the -OH group.

[0129] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0130] The term "amino" refers to -NH2.

[0131] The term "cyano" refers to -CN.

[0132] The term "nitro" refers to -NO2.

[0133] The term "oxo" refers to =O.

[0134] The term "thio-" refers to =S.

[0135] The term "carboxyl group" refers to -C(O)OH.

[0136] The term "thiol" refers to -SH.

[0137] The term "ester group" refers to -C(O)O (alkyl) or -C(O)O (cycloalkyl), where alkyl and cycloalkyl are as defined above.

[0138] The term "acyl" refers to a compound containing a -C(O)R group, where R is an alkyl, cycloalkyl, heterocyclic, aryl, or heteroaryl group as defined above.

[0139] The term "sulfonyl" refers to a compound containing a -S(O)2R group, where R is an alkyl, cycloalkyl, heterocyclic, aryl, or heteroaryl group as defined above.

[0140] The compounds of this invention can be in deuterated form. Each available hydrogen atom bonded to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize the deuterated form of the compounds by referring to relevant literature. Commercially available deuterated starting materials can be used to prepare the deuterated form of the compounds, or they can be synthesized using conventional techniques with deuterating reagents.

[0141] "Optional" or "optionally" means that the event or environment described below may but does not have to occur, and the description includes the possibility or absence of such event or environment. For example, "optionally alkyl-substituted heterocyclic group" means that the alkyl group may but does not have to be present, and the description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

[0142] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, and more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0143] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0144] "Medicinal salts" refers to salts of the compounds of this invention that are safe and effective when used in mammals and have the appropriate biological activity.

[0145] Method for synthesizing the compounds of the present invention

[0146] The compound represented by general formula (IV) of this invention can be prepared by the following scheme 1.

[0147]

[0148] Option 1

[0149] Step 1: Under alkaline conditions, compound IVa reacts with R 1 L undergoes a substitution reaction to give compound IVb, wherein the alkaline conditions are preferably potassium carbonate; wherein L is a halogen, preferably fluorine, chlorine, or bromine, and more preferably bromine;

[0150] Step 2: Under alkaline conditions, compound IVb undergoes a methylation reaction with iodomethane to yield compound IVc, wherein the preferred alkaline conditions are lithium diisopropylamino.

[0151] Step 3: Under alkaline conditions, compound IVcN,N-dimethylformamide undergoes a substitution reaction to give compound IVd, wherein the alkaline conditions are preferably n-butyllithium;

[0152] Step 4: Under alkaline conditions, compound IVd undergoes a dehydration reaction with hydroxylamine hydrochloride to obtain compound IVe, wherein the preferred alkaline conditions are sodium acetate;

[0153] Step 5: Under catalytic conditions, compound IVe and IVk undergo a cyclization reaction to yield compound IVf, wherein the catalyst is preferably N-chlorosuccinimide (NCS);

[0154] Step 6: Under catalytic conditions, compound IVf undergoes a substitution reaction with N-bromosuccinimide (NBS) to give compound IVg, wherein the catalyst is preferably azobisisobutyronitrile (AIBN);

[0155] Step 7: Under alkaline conditions, compound IVg undergoes ammonolysis and cyclization to give compound IVh, wherein the preferred alkaline conditions are ammonia water (25%).

[0156] Step 8: Under the conditions of a thioreagent, compound IVh undergoes a thioreaction to give compound IVi, wherein the thioreagent is preferably diphosphine pentasulfide;

[0157] Step 9: Under the conditions of the methylating reagent, compound IVi undergoes a methylation reaction to obtain compound IVj, wherein the methylating reagent is trimethyloxonium tetrafluoroboric acid;

[0158] Step 10: Under catalytic conditions, compound IVj undergoes a cyclization reaction to give the compound represented by general formula (IV), wherein the catalyst is preferably glacial acetic acid;

[0159] Where: X1, X2, R 1 R 2 R 3 R 5 and R 6 As defined by general formula (IV). Attached Figure Description

[0160] Figure 1A-1C Tumor growth curves of BaF3 KIF5B RET-G810R cell subcutaneous xenograft tumor model mice after administration of the compound; Figure 1A The tumor growth curve of the compound in Example 4 is shown. Figure 1B The tumor growth curve of the compound in Example 15 is shown. Figure 1C The tumor growth curve is for the compound in Example 19.

[0161] Figure 2A-2C The body weight curve of BaF3 KIF5B RET-G810R cell subcutaneous xenograft tumor model mice during drug administration; Figure 2A The weight curve of the compound in Example 4 is shown. Figure 2B The body weight curve of compound 15 is shown below. Figure 2C The weight curve of the compound in Example 19 is shown. Detailed Implementation

[0162] Further examples are provided to illustrate the compounds of the present invention and their preparation, and these examples demonstrate methods for preparing or using said compounds. However, it is to be understood that these examples do not limit the scope of the invention. Variations of the invention now known or further developed are considered to fall within the scope of the invention described and claimed herein.

[0163] The compounds of this invention are prepared using convenient starting materials and common preparation steps. Typical or preferred reaction conditions are provided, such as reaction temperature, time, solvent, pressure, and molar ratio of reactants. However, unless otherwise specified, other reaction conditions may also be adopted. Optimal conditions may vary depending on the specific reactants or solvents used, but in general, the optimal reaction steps and conditions can be determined.

[0164] In addition, this invention may employ protecting groups to protect certain functional groups from unwanted reactions. Suitable protecting groups for various functional groups and their protection or deprotection conditions are well known to those skilled in the art. For example, TW Greene and GMWuts' "Protecting Groups in Organic Preparations" (3rd edition, Wiley, New York, 1999 and cited references therein) provides a detailed description of the protection or deprotection of numerous protecting groups.

[0165] The separation and purification of compounds and intermediates are carried out using appropriate methods and procedures depending on specific needs, such as filtration, extraction, distillation, crystallization, column chromatography, preparative thin-layer chromatography, preparative high-performance liquid chromatography, or a combination of the above methods. Specific methods can be found in the examples described in this invention. Of course, other similar separation and purification methods can also be used. Conventional methods (including physical constants and spectroscopic data) can be used for characterization.

[0166] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts were expressed in 10⁻¹⁰. -6 The unit (ppm) is given. NMR measurements were performed using a Brukerdps 300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0167] MS measurements were performed using an LC (Agilent 1260 Infinity) / MS (G6125B) mass spectrometer (manufacturer: Agilent).

[0168] Preparative liquid chromatography was performed using an lc6000 high-performance liquid chromatograph (manufacturer: Innovation Tongheng). The chromatographic column was a Daisogel C18 10μm 100A (30mm × 250mm), and the mobile phase was acetonitrile / water.

[0169] Thin-layer chromatography (TLC) uses Qingdao Ocean Chemical GF254 silica gel plates. The silica gel plates used for reaction monitoring in TLC have a diameter of 0.20 mm to 0.25 mm, while those used for separation and purification have a diameter of 0.5 mm.

[0170] Silica gel column chromatography uses Qingdao marine silica gel of 100-200 mesh, 200-300 mesh, and 300-400 mesh as carriers.

[0171] The known starting materials of this invention can be synthesized using or according to methods known in the art, or can be purchased from companies such as NetEase, Beijing Coupling, Sigma, Bailingwei, Yishiming, Shanghai Shuya, Shanghai Inokai, Anaiji Chemical, Shanghai Bide, and Nanjing Yaoshi.

[0172] Unless otherwise specified in the examples, all reactions can be carried out under a nitrogen atmosphere.

[0173] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0174] The terms "reaction solvent," "organic solvent," or "inert solvent" are each used to indicate that the solvent does not participate in the reaction under the described reaction conditions. These include solvents such as benzene, toluene, acetonitrile, tetrahydrofuran (THF), dimethylformamide (DMF), chloroform, dichloromethane, diethyl ether, methanol, N-methylpyrrolidone (NMP), and pyridine. Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0175] The chemical reactions described in this invention are generally carried out under normal pressure. The reaction time and conditions are, for example, at one atmosphere, between -78°C and 200°C, and are completed in approximately 1 to 24 hours. If the reaction is carried out overnight, the reaction time is generally 16 hours. Unless otherwise specified in the examples, the reaction temperature is room temperature, between 20°C and 30°C.

[0176] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent systems used in the reactions were: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system, and C: acetone. The volume ratio of the solvents was adjusted according to the polarity of the compounds.

[0177] The eluent system for column chromatography and the developing solvent system for thin-layer chromatography used to purify the compound include: A: dichloromethane and methanol system, B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound. Small amounts of basic or acidic reagents such as triethylamine and trifluoroacetic acid can also be added for adjustment.

[0178] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be used in the methods of this invention.

[0179] Example

[0180] Example 1: Preparation of 3-cyclopropyl-9-isopropyl-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (1)

[0181]

[0182] Step 1: Preparation of 5-bromo-4-chloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine (1a)

[0183] At room temperature, 5-bromo-4-chloro-7H-pyrrolo[2,3-d]pyrimidine (5.00 g, 21.5 mmol) was dissolved in DMF (50 mL), and cesium carbonate (10.5 g, 32.3 mmol) and 2-bromopropane (3.20 g, 25.8 mmol) were added. The reaction mixture was heated to 60 °C and stirred overnight. The mixture was quenched with 100 mL of ice water, extracted with ethyl acetate (EA) (100 mL x 2), washed with saturated NaCl (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 100:1-10:1) to give 5.12 g of the title compound as a white solid, yield: 86.7%.

[0184] LC-MS: m / z 274 [M+H] + .

[0185] Step 2: Preparation of 5-bromo-4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine (1b)

[0186] At room temperature, 5-bromo-4-chloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidine (1a) (5.00 g, 18.3 mmol) was dissolved in 50 mL of THF. Under a nitrogen atmosphere at -78 °C, lithium diisopropylamino (LDA) (18.0 mL, 2 M, 36.6 mmol) was slowly added dropwise using a syringe, and stirring continued for 30 minutes. CH3I (7.80 g, 55 mmol) was then added dropwise, and the mixture was slowly brought to room temperature. The mixture was quenched with 50 mL of water, extracted with EA (100 mL x 2), washed with saturated NaCl (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: PE / EA = 100:1-10:1) to give 4.52 g of the title compound as a white solid, yield: 86.1%.

[0187] LC-MS: m / z 287 [M+H] + .

[0188] Step 3: Preparation of 4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde (1c)

[0189] At room temperature, 5-bromo-4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine (1b) (4.00 g, 14.0 mmol) was dissolved in 50 mL of THF. Under a nitrogen atmosphere at -50 °C, n-BuLi (11.2 mL, 2.5 M, 28.0 mmol) was slowly added dropwise using a syringe, and stirring continued for 30 minutes. DMF (10.2 g, 70.0 mmol) was then added dropwise, and the mixture was slowly brought to room temperature. The solution was quenched with 50 mL of water, extracted with EA (100 mL x 2), washed with saturated NaCl (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: PE / EA = 50:1–8:1) to give 1.6 g of the title compound as a white solid, yield: 48.5%.

[0190] LC-MS: m / z 238 [M+H] + .

[0191] Step 4: Preparation of 4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde oxime (1d)

[0192] 4-Chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde (1c) (1.3 g, 5.5 mmol) was dissolved in 20 mL of EtOH at room temperature. NaOAc (1.13 g, 13.7 mmol) and hydroxylamine hydrochloride (0.95 g, 13.7 mmol) were added, and the mixture was stirred at room temperature for 4 h. 20 mL of water was added, and the mixture was extracted with EA (20 mL x 2), washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: PE / EA = 50:1-5:1) to give 1.22 g of the title compound as a white solid, yield: 87.0%.

[0193] LC-MS: m / z 253 [M+H] + .

[0194] Step 5: Preparation of 3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole (1e)

[0195] 4-Chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde oxime (1d) (1.80 g, 7.10 mmol) was dissolved in 20 mL of DMF at room temperature. Potassium bicarbonate (1.40 g, 14.2 mmol), N-chlorosuccinimide (NCS) (1.24 g, 9.23 mmol), and ethynylcyclopropane (1.40 g, 21.3 mmol) were added, and the mixture was stirred overnight at room temperature. 20 mL of water was added, and the mixture was extracted with EA (20 mL x 2), washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: PE / EA = 100:1-10:1) to give 1.42 g of the title compound as a white solid, yield: 63.1%.

[0196] LC-MS: m / z 317 [M+H] + .

[0197] Step 6: Preparation of 3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropyl-4-iodoisoxazole (1f)

[0198] 3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole (1e) (1.32 g, 4.18 mmol) was dissolved in 20 mL of acetonitrile (ACN) at room temperature. Trifluoroacetic acid (TFA) (1.43 g, 12.5 mmol) and N-iodosuccinimide (NIS) (1.41 g, 6.27 mmol) were added, and the mixture was stirred overnight at room temperature. 20 mL of water was added, and the mixture was extracted with EA (20 mL x 2), washed with saturated brine (30 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: PE / EA = 100:1-10:1) to give 1.45 g of the title compound as a reddish-brown solid, yield: 78.8%.

[0199] LC-MS: m / z 442 [M+H] + .

[0200] Step 7: Preparation of methyl 3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid (1 g)

[0201] At room temperature, 3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropyl-4-iodoisoxazole (1f) (3.20 g, 7.23 mmol) was dissolved in THF (5 mL). At 0 °C, isopropyl magnesium chloride and lithium chloride complex salt (7.23 mL, 1.3 M, 9.40 mmol) were added, and stirring was continued for 0.5 hours. Methyl chloroformate (1.36 g, 14.48 mmol) was added, and the mixture was brought to room temperature and stirred overnight. Quenching with 10 mL of ice water, extraction with EA (20 mL x 2), washing with saturated NaCl (20 mL x 2), drying with anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and separating and purifying the residue by silica gel column chromatography (mobile phase: PE / EA = 100:1-10:1) to give 1.52 g of the title compound as a white solid, yield: 56.3%.

[0202] LC-MS: m / z 375 [M+H] + .

[0203] Step 8: Preparation of methyl 3-(6-(bromomethyl)-4-chloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid (1h)

[0204] At room temperature, methyl 3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazol-4-carboxylate (1 g) (1.20 g, 3.21 mmol) was dissolved in 20 mL of dichloroethane (DCE), and NBS (0.68 g, 3.85 mmol) and azobisisobutyronitrile (AIBN) (53.0 mg, 3.85 mmol) were added. The reaction mixture was heated to 80 °C and stirred overnight. The reaction mixture was quenched with 40 mL of water, extracted with EA (50 mL x 2), washed with saturated NaCl (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 1.6 g of crude title compound as a white solid.

[0205] LC-MS: m / z 453 [M+H] + .

[0206] Step 9: Preparation of 11-amino-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (1i)

[0207] At room temperature, crude methyl 3-(6-(bromomethyl)-4-chloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid (1 h) (1.50 g, 3.32 mmol) was dissolved in 15 mL of 1,4-dioxane, and 15 mL of ammonia (25%) was added. The reaction mixture was heated to 90 °C and stirred for 24 h. The reaction was quenched with 50 mL of water, extracted with EA (100 mL x 2), washed with saturated NaCl (100 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 1:1-1:3) to give 450 mg of the title compound as a white solid, yield: 40.9%.

[0208] LC-MS: m / z 339 [M+H] + .

[0209] Step 10: Preparation of 11-amino-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-thione (1j)

[0210] At room temperature, 11-amino-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (1i) (50.0 mg, 0.150 mmol) was dissolved in 1 mL of ACN, and triethylamine (30.0 mg, 0.300 mmol) and diphosphine pentasulfide (42.0 mg, 0.220 mmol) were added. The reaction mixture was heated to 80 °C and stirred overnight. The reaction mixture was concentrated under reduced pressure, and the residue was separated by preparative high-performance liquid chromatography (HPLC) (column: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.1% FA), gradient: 10%-50%), yielding 18 mg of the title compound as a white solid, yield: 34.6%.

[0211] LC-MS: m / z 354 [M+H] + .

[0212] Step 11: Preparation of 3-cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k)

[0213] At room temperature, 11-amino-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-thione (1j) (60.0 mg, 0.170 mmol) was dissolved in DCM (5 mL). Trimethyloxonium tetrafluoroboric acid (50.0 mg, 0.340 mmol) was added at 0 °C, and the mixture was stirred overnight at room temperature. The mixture was quenched with 10 mL of ice water, extracted with DCM (20 mL x 2), washed with saturated NaCl (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 50 mg of the title compound as a white solid, crude product.

[0214] LC-MS: m / z 369 [M+H] + .

[0215] Step 12: Preparation of 3-cyclopropyl-N-(2,2-dimethoxyethyl)-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4,11-diamine (1l)

[0216] At room temperature, 3-cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (50.0 mg, 0.136 mmol) was dissolved in 1 mL of EtOH, and 2,2-dimethoxyethane-1-amine (28.0 mg, 0.271 mmol) was added. The reaction mixture was heated to 80 °C and stirred overnight. The reaction mixture was quenched with 10 mL of water, extracted with EA (20 mL x 2), washed with saturated NaCl (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 60 mg of the title compound as a white solid, crude product.

[0217] LC-MS: m / z 426 [M+H] + .

[0218] Step 13: Preparation of 3-cyclopropyl-9-isopropyl-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (1)

[0219] At room temperature, crude 3-cyclopropyl-N-(2,2-dimethoxyethyl)-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4,11-diamine (1 L) (60.0 mg, 0.141 mmol) was dissolved in 1 mL of acetic acid, and the reaction mixture was heated to 100 °C and stirred overnight. The reaction mixture was concentrated under reduced pressure, and the residue was separated by preparative liquid chromatography (HPLC) (column: Daisogei 30 mm × 250 mm, C18, 10 μm 100A; mobile phase: acetonitrile / water (0.1% FA); gradient: 20%–28%), yielding 6 mg of the title compound as a white solid, yield: 11.7%.

[0220] LC-MS: m / z 362 [M+H] + .

[0221] 1 H NMR (400MHz, DMSO-d6) δ8.16(s,1H),7.56(d,J=1.2Hz,1H),7.45(s,2H),7.08(d,J=1.2Hz,1H),5.58(s ,2H),5.28(h,J=6.9,6.2Hz,1H),2.99(tt,J=7.9,5.6Hz,1H),1.65(d,J=6.9Hz,6H),1.31-1.18(m,4H).

[0222] Example 2: Preparation of 3-cyclopropyl-9-isopropyl-6-methyl-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]-azacycloheptane-13-amine (2)

[0223]

[0224] Step 1: Preparation of 3-cyclopropyl-7-isopropyl-4-methoxy-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (2a)

[0225] At room temperature, 11-amino-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (1i) (100 mg, 0.296 mmol) was dissolved in DCM (5 mL). Trimethyloxonium tetrafluoroboric acid (87.6 mg, 0.592 mmol) was added at 0 °C, and the mixture was stirred overnight at room temperature. The mixture was quenched with 10 mL of ice water, extracted with DCM (20 mL x 2), washed with saturated NaCl (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 94 mg of the title compound as a white solid, crude product.

[0226] LC-MS: m / z 353 [M+H] + .

[0227] Step 2: Preparation of 3-cyclopropyl-9-isopropyl-6-methyl-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]-azacycloheptane-13-amine (2)

[0228] At room temperature, 3-cyclopropyl-7-isopropyl-4-methoxy-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (2a) (30.0 mg, 0.085 mmol) was dissolved in 1 mL of EtOH, and acetylhydrazine (13.0 mg, 0.170 mmol) was added. The mixture was heated to 90 °C and stirred for 48 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated by preparative high-performance liquid chromatography (HPLC) (column: Daisogei 30 mm × 250 mm, C18, 10 μm 100A; mobile phase: acetonitrile / water (0.1% FA); gradient: 10%–40%), yielding 4.1 mg of the title compound as a white solid, yield: 12.8%.

[0229] LC-MS: m / z 377 [M+H] + .

[0230] 1 H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.44(s,2H),5.46(s,2H),5.18(p,J=7.0Hz,1H),2.9 3(tt,J=8.1,5.4Hz,1H),2.57(s,3H),1.68(d,J=6.8Hz,6H),1.30(tt,J=8.1,2.9Hz,4H).

[0231] Example 3: Preparation of 3-cyclopropyl-9-isopropyl-6-methyl-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (3)

[0232]

[0233]

[0234] Step 1: Preparation of 3-cyclopropyl-9-isopropyl-6-methyl-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (3)

[0235] Azacycloheptan-compound 1k (50 mg, 0.135 mmol) was dissolved in 2 mL of N-methylpyrrolidone (NMP), and propargylamine (14.9 mg, 0.271 mmol) and p-toluenesulfonic acid (2.30 mg, 0.0130 mmol) were added. The mixture was microwave-stirred at 150 °C for 50 min. The solution was then separated directly by high-performance liquid chromatography (HPLC) (column: Daisogei 30 mm * 250 mm, C18, 10 μm 100A; mobile phase: acetonitrile / water (0.1% FA); gradient: 20%–28%, 254 nM) to give 5 mg of the title compound as a white solid, yield: 9.8%.

[0236] LC-MS: m / z 375.10 [M+H] + .

[0237] 1H NMR (400MHz, DMSO-d6) δ8.16 (s, 1H), 7.42 (s, 2H), 6.85 (d, J = 1.2Hz, 1H), 5.34 (s, 2H), 5.22-5.10 (m ,1H),2.96(tt,J=7.6,5.9Hz,1H),2.38(d,J=1.1Hz,3H),1.68(d,J=6.8Hz,6H),1.27-1.19(m,4H).

[0238] Example 4: Preparation of 3-cyclopropyl-9-isopropyl-8,9-dihydroisooxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]tetrazo[1,5-a]azacycloheptane-13-amine (4)

[0239]

[0240] Step 1: Preparation of benzyl 3-cyclopropylpropynate (4a)

[0241] Under a nitrogen atmosphere, n-butyllithium (2.5M n-hexane solution, 6.67 mL, 16.7 mmol) was added to a tetrahydrofuran (10 mL) solution of ethynylcyclopropane (1.00 g, 15.1 mmol) at -78 °C. The mixture was stirred for 1 hour, and then benzyl chloroformate (2.84 g, 16.6 mmol) was added. The reaction mixture was slowly heated to -10 °C, quenched with water (15 mL), and extracted with ethyl acetate (3 x 20 mL). The organic phases were combined, washed with saturated brine (1 x 50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 3:1) to give 2.20 g of the title compound as a white solid, yield: 69.0%.

[0242] LC-MS: m / z 201 [M+H] + .

[0243] Step 2: Preparation of 3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (4b)

[0244] Under a nitrogen atmosphere, potassium bicarbonate (0.870 g, 8.70 mmol), N-chlorosuccinimide (NCS) (0.760 g, 5.66 mmol), and benzyl 3-cyclopropylpropynate (1.74 g, 8.71 mmol) were added to a solution of 4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde oxime (1d) (1.10 g, 4.35 mmol) in N,N-dimethylformamide (15 mL) at room temperature. The mixture was stirred overnight at room temperature. The reaction solution was diluted with water (30 mL) and extracted with ethyl acetate (3 x 35 mL). The organic phases were combined, washed with saturated brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 5:1) to give 1.415 g of the title compound as a white solid, yield: 68.5%.

[0245] LC-MS: m / z 451 [M+H] + .

[0246] Step 3: Preparation of 3-(6-(bromomethyl)-4-chloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (4c)

[0247] Under a nitrogen atmosphere, N-bromosuccinimide (NBS) (0.89 g, 4.99 mmol) and azobisisobutyronitrile (AIBN) (0.220 g, 1.33 mmol) were added to a solution of compound 4b (1.50 g, 3.33 mmol) in 1,2-dichloroethane (30 mL), and the mixture was stirred overnight at room temperature. The reaction solution was diluted with water (50 mL) and extracted with dichloromethane (3 x 60 mL). The combined organic phases were washed with saturated brine (1 x 200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 6:1) to give 1.14 g of the title compound as a colorless, transparent oil, yield: 61.6%.

[0248] LC-MS: m / z 529 [M+H] + .

[0249] Step 4: Preparation of 11-amino-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (1i)

[0250] Under a nitrogen atmosphere, compound 4c (570 mg, 1.08 mmol, 1.00 equiv) was added to a mixed solution of 1,4-dioxane (10 mL) and ammonia (6 mL) at room temperature. The mixture was heated to 85 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: dichloromethane / methanol = 99:1-93:7) to give 200 mg of the title compound as a white solid, yield: 52.2%.

[0251] LC-MS: m / z 339 [M+H] + .

[0252] Step 5: Preparation of 11-amino-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-thione (1j)

[0253] Under a nitrogen atmosphere, phosphorus pentasulfide (199 mg, 0.895 mmol) was added to a solution of 202 mg (0.597 mmol) of aziridine-compound 1i in acetonitrile (5 mL) at room temperature. The reaction was heated to 80 °C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: dichloromethane / methanol = 99:1-90:10) to give 140 mg of the title compound as a light green solid, in 60.6% yield.

[0254] LC-MS: m / z 355 [M+H] + .

[0255] Step 6: Preparation of 3-cyclopropyl-9-isopropyl-8,9-dihydroisooxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]tetrazo[1,5-a]azacycloheptane-13-amine (4)

[0256] Under a nitrogen atmosphere, at room temperature, azidotrimethylsilane (58.5 mg, 0.508 mmol) was added to a solution of azidocycloheptan-compound 1j (90.0 mg, 0.254 mmol) in dichloromethane (5 mL), and the mixture was stirred for 10 minutes. Ferric chloride (49.4 mg, 0.305 mmol) was then added, and the mixture was stirred at room temperature for 2 days. The reaction mixture was filtered, and the filter cake was washed with acetonitrile (3 x 15 mL). The filtrate was concentrated under reduced pressure. The residue was separated by high-performance liquid chromatography (HPLC) (column: XBridge Prep OBD C18 Column, 19 x 250 mm, 5 μm; mobile phase A: water (10 mmol / L ammonium bicarbonate + 0.1% ammonia), mobile phase B: methanol; flow rate: 60 mL / min; gradient: 20% B to 50% B for 8 minutes) to give 20.8 mg of the title compound as a white solid, in a yield of 22.14%.

[0257] LC-MS: m / z 364 [M+H] + .

[0258] 1 H NMR(400MHz,DMSO-d6)δ8.18(s,1H),7.52(s,2H),6.18(s,2H),5.32-5.22(m,1H), 2.96-2.86(m,1H),1.67(d,J=6.7Hz,6H),1.37(dtd,J=13.3,8.2,6.7,4.0Hz,4H).

[0259] Example 5: Preparation of 3-cyclopropyl-9-isopropyl-8,9-dihydroisoxazo[4”,3”:6',7']pyrido[3”,2”:4',5']cycloheptano[1',2':4,5]pyrrolo[2,3-d]pyrimidine-13-amine (5)

[0260]

[0261] Step 1: Preparation of 1-(3-(4-chloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazol-4-yl)aceto-1-one (5a)

[0262] Under a nitrogen atmosphere, at -78°C, a tetrahydrofuran solution (1.3 M, 3.2 mL, 4.13 mmol) of the isopropyl magnesium chloride-lithium chloride complex salt was added to a tetrahydrofuran solution (15 mL). After stirring for 30 minutes, acetic anhydride (484 mg, 4.74 mmol) was added dropwise. The mixture was allowed to rise naturally to room temperature and stirred for 2 hours. The reaction mixture was quenched with water (20 mL) and extracted with ethyl acetate (3 x 30 mL). The organic phases were combined, washed with saturated brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 4:1) to give 900 mg of the title compound as a pale yellow solid, yield: 75.3%.

[0263] LC-MS: m / z 359 [M+H] + .

[0264] Step 2: Preparation of 1-(3-(6-(bromomethyl)-4-chloro-7-isopropyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazol-4-yl)-1-one (5b)

[0265] Under a nitrogen atmosphere, N-bromosuccinimide (595 mg, 3.34 mmol) and azobisisobutyronitrile (73.2 mg, 0.446 mmol) were added to a solution of compound 5a (800 mg, 2.23 mmol) in 1,2-dichloroethane (16 mL), and the mixture was stirred overnight at room temperature. The reaction mixture was diluted with water (30 mL) and extracted with dichloromethane (3 x 40 mL). The combined organic phases were washed with saturated brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative chromatography (mobile phase: petroleum ether / ethyl acetate = 6:1) to give 228 mg of the title compound as a colorless, transparent oil, in yield: 82.8%.

[0266] LC-MS: m / z 437 [M+H] + .

[0267] Step 3: Preparation of 11-chloro-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4”,3”:6',7']cycloheptano[1',2':4,5]pyrrolo[2,3-d]pyrimidin-4-(5H)-1-one (5c)

[0268] Under a nitrogen atmosphere, lithium bis(trimethylsilylamino)amine (1M in THF, 3.40 mL, 3.42 mmol) was added to a tetrahydrofuran (20 mL) solution of compound 5b (750 mg, 1.71 mmol) at -78 °C. The mixture was stirred for 30 minutes and then slowly brought to room temperature. The mixture was quenched with water (20 mL). The aqueous phase was extracted with ethyl acetate (3 x 30 mL). The organic phases were combined, washed with saturated brine (1 x 100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 5:1) to give 250 mg of the title compound as a pale yellow oil, yield: 38.9%.

[0269] LC-MS: m / z 357 [M+H] + .

[0270] Step 4: Preparation of 11-(bis(4-methoxybenzyl)amino)-3-cyclopropyl-7-isopropyl-6,7-dihydroisoxazolo[4”,3”:6',7']cycloheptano[1',2':4,5]pyrrolo[2,3-d]pyrimidin-4(5H)-one (5d)

[0271] Under a nitrogen atmosphere, bis(4-methoxybenzyl)amine (721 mg, 2.81 mmol) and N,N-diisopropylethylamine (725 mg, 5.61 mmol) were added to a 4 mL solution of compound 5c (200 mg, 0.561 mmol) in dimethyl sulfoxide at room temperature. The reaction mixture was heated to 95 °C and stirred for 1 hour. The resulting residue was purified by C18 silica gel column chromatography (mobile phase: water and acetonitrile, 10% to 80% gradient, 30 min) to give 200 mg of the title compound as a yellow solid, in 58.7% yield.

[0272] LC-MS: m / z 578 [M+H] + .

[0273] Step 5: Preparation of 3-cyclopropyl-9-isopropyl-N,N-bis(4-methoxybenzyl)-8,9-dihydroisoxazo[4”,3”:6’,7’]pyrido[3”,2”:4’,5’]cycloheptano[1’,2’:4,5]pyrrolo[2,3-d]pyrimidine-13-amine (5e)

[0274] Under an oxygen atmosphere, at room temperature, 4-methylbenzene-1-sulfonic acid hydrate (9.88 mg, 0.052 mmol) and copper bis(trifluoromethanesulfonic acid) (3.13 mg, 0.009 mmol) were added fractionally to a hexanol (2 mL) solution of compound 5d (50.0 mg, 0.087 mmol) and propane-1,3-diamine (64.2 mg, 0.870 mmol). The reaction solution was heated to 130 °C and stirred overnight. The reaction solution was diluted with water (10 mL). Extracted with ethyl acetate (3 x 10 mL), the organic phases were combined, washed with saturated brine (1 x 20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative chromatography (mobile phase: dichloromethane / methanol = 20:1) to give 23 mg of the title compound as a brownish-yellow solid, yield: 34.7%.

[0275] LC-MS: m / z 613 [M+H] + .

[0276] Step 6: Preparation of 3-cyclopropyl-9-isopropyl-8,9-dihydroisoxazo[4”,3”:6',7']pyrido[3”,2”:4',5']cycloheptano[1',2':4,5]pyrrolo[2,3-d]pyrimidine-13-amine (5)

[0277] Under a nitrogen atmosphere, trifluoroacetic acid (0.3 mL) and 3 drops of p-toluenesulfonic acid were added to a solution of compound 5e (55.0 mg, 0.090 mmol) in dichloromethane (3 mL) at 0 °C. The reaction mixture was then stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure. The resulting residue was purified by high-performance liquid chromatography (HPLC) (column: Xselect SCH C18OBD, 30*150 mm, 5 μm; mobile phase A: water (10 mmol / L ammonium bicarbonate + 0.1% ammonia), mobile phase B: acetonitrile; flow rate: 60 mL / min; gradient: 35% B to 65% B for 8 min) to give 8.1 mg of the title compound as a white solid, in 24.2% yield.

[0278] LC-MS: m / z 373 [M+H] + .

[0279] 1H NMR (400MHz, DMSO-d6) δ8.58(d,J=4.7Hz,1H),8.13(s,1H),8.05(d,J=7.6Hz,1H),7.44-7.37(m,1H),7.2 6(s,2H),5.31(d,J=10.4Hz,1H),4.21(s,2H),3.13(s,1H),1.66(d,J=6.8Hz,6H),1.26(d,J=6.9Hz,4H).

[0280] Example 6: Preparation of 3-cyclopropyl-9-(1,3-difluoropropane-2-yl)-6-methyl-8,9-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]-azacycloheptane-13-amine (6)

[0281]

[0282] Step 1: Preparation of benzyl 3-(4-chloro-7-(1,3-difluoroprop-2-yl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid ester (6a)

[0283] At room temperature, 3-(4-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazol-4-carboxylic acid benzyl ester (1.50 g, 3.67 mmol), 1,3-difluoro-2-propanol (0.705 g, 7.35 mmol), and triphenylphosphine (0.632 g, 5.50 mmol) were dissolved in THF (20 mL). Diisopropyl azodicarbonate (DIAD) (1.11 g, 5.50 mol) was added at 0 °C, and the mixture was stirred overnight at 50 °C. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 5) to give 1.6 g of the title compound as a pale yellow solid, yield: 89.9%.

[0284] LC-MS: m / z 487.13 [M+H] + .

[0285] Step 2: Preparation of benzyl 3-(6-(bromomethyl)-4-chloro-7-(1,3-difluoroprop-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid ester (6b)

[0286] At room temperature, benzyl 3-(4-chloro-7-(1,3-difluoroprop-2-yl)-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid ester (1.60 g, 3.29 mmol), NBS (0.879 g, 4.93 mmol), and azobisisobutyronitrile (AIBN) (107 mg, 0.658 mmol) were dissolved in DCE (20 mL). The mixture was stirred overnight at 50 °C. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 5) to give 1.60 g of the title compound as a pale yellow solid, yield: 86.4%.

[0287] LC-MS: m / z 565.04 [M+H] + .

[0288] Step 3: Preparation of 11-amino-3-cyclopropyl-7-(1,3-difluoroprop-2-yl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4-(5H)-one (6c)

[0289] At room temperature, benzyl 3-(6-(bromomethyl)-4-chloro-7-(1,3-difluoroprop-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid ester (160 mg, 2.83 mmol) was dissolved in dioxane (20 mL), and ammonia water (20 mL) was added. The mixture was stirred overnight at 90 °C. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 10 / 1) to give 300 mg of the title compound as a brown solid, yield: 28.3%.

[0290] LC-MS: m / z 375.13 [M+H] + .

[0291] Step 4: Preparation of 11-amino-3-cyclopropyl-7-(1,3-difluoropropyl-2-yl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4-(5H)-thione (6d)

[0292] At room temperature, 300 mg (0.802 mmol) of 11-amino-3-cyclopropyl-7-(1,3-difluoropropyl-2-yl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4-(5H)-one was dissolved in 20 mL of ACN. Triethylamine (162 mg, 1.60 mmol) and diphosphine pentasulfide (355 mg, 1.60 mmol) were added sequentially, and the mixture was stirred overnight at 80 °C. The mixture was quenched with 20 mL of ice water, filtered, and the filter cake was washed twice with methanol and dried to give 200 mg of the title compound as a gray solid. Yield: 63.8%.

[0293] LC-MS: m / z 391.11 [M+H] + .

[0294] Step 5: Preparation of 3-cyclopropyl-7-(1,3-difluoropropyl-2-yl)-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (6e)

[0295] At room temperature, 200 mg (0.789 mmol) of 11-amino-3-cyclopropyl-7-(1,3-difluoropropyl-2-yl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4-(5H)-thione was dissolved in 5 mL of DCM. Trimethyloxonium tetrafluoroboric acid (233 mg, 1.57 mmol) was added at 0 °C, and the reaction mixture was stirred overnight at room temperature. The reaction was quenched with 10 mL of saturated ammonium chloride solution, extracted with 20 mL of DCM, washed twice with 20 mL of saturated NaCl, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 10 / 1) to give 80 mg of the title compound as a brown solid, yield: 83.6%.

[0296] LC-MS: m / z 405.12 [M+H] + .

[0297] Step 6: Preparation of 3-cyclopropyl-9-(1,3-difluoropropyl-2-yl)-6-methyl-8,9-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (6)

[0298] At room temperature, 3-cyclopropyl-7-(1,3-difluoroprop-2-yl)-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidine[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (30.0 mg, 0.0742 mmol) was dissolved in 1 mL of EtOH, and acetylhydrazine (10.1 mg, 0.148 mmol) was added. The mixture was stirred at 90 °C for 2 days. The reaction solution was concentrated under reduced pressure, and the residue was separated by preparative high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.1% FA), gradient: 10%-40%), yielding 3 mg of the title compound as a white solid, yield: 10.0%.

[0299] LC-MS: m / z 413.16 [M+H] + .

[0300] 1 H NMR(400MHz,DMSO-d6)δ8.42(s,1H),8.18(s,1H),7.55(s,2H),5.49(s,2H),5.36-5.21(m,1H),5.18-5.09 (m,2H),5.03(dd,J=9.6,4.9Hz,1H),2.92(tt,J=7.9,5.5Hz,1H),2.54(s,3H),1.31(dd,J=9.7,3.4Hz,4H).

[0301] Example 7: Preparation of 9-cyclopentyl-3-cyclopropyl-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (7)

[0302]

[0303] Step 1: Preparation of 4-chloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (7a)

[0304] At room temperature, 12.0 g (0.0784 mol) of 4-chloro-7H-pyrrolo[2,3-d]pyrimidine was dissolved in 100 mL of DMF. NaH (60% in oil, 2.25 g, 0.0941 mol) was added at 0 °C, and stirring was continued for half an hour. Then, 15.6 g (0.0941 mol) of 2-(trimethylsilyl)ethoxymethyl chloride was added, and the mixture was stirred at room temperature for 4 hours. The mixture was quenched with 200 mL of ice water, extracted three times with EA, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 20) to give 19.0 g of the title compound as a pale yellow oil, yield: 85.6%.

[0305] LC-MS: m / z 284.09 [M+H] + .

[0306] Step 2: Preparation of 4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine (7b)

[0307] At room temperature, 19.0 g (0.0671 mol) of 4-chloro-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine was dissolved in 200 mL of THF. LDA (2.5 N, 26.8 mL, 0.0671 mol) was added dropwise at -78 °C, and stirring was continued for 0.5 hours. CH3I (47.6 g, 0.335 mol) was then added dropwise. The mixture was brought to room temperature and stirred overnight. The reaction solution was concentrated under reduced pressure, quenched with 100 mL of ice water, extracted three times with EA, dried over anhydrous ammonium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 20) to give 17.2 g of the title compound as a yellow solid, yield: 86.0%.

[0308] LC-MS: m / z 298.11 [M+H] + .

[0309] Step 3: Preparation of 4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde (7c).

[0310] At room temperature, 17.2 g (0.0579 mol) of 4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine was dissolved in 150 mL of DMF, and 44.0 g (0.289 mmol) of POCl3 was added. The reaction mixture was heated to 70 °C and stirred for 4 h. After cooling to room temperature, 300 mL of ice water was added to quench the reaction, and the mixture was extracted three times with EA. The extract was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 10) to give 12.5 g of the title compound as a yellow solid, yield: 66.4%.

[0311] LC-MS: m / z 326.10 [M+H] + .

[0312] Step 4: Preparation of (E)-4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde oxime (7d)

[0313] At room temperature, 12.5 g (0.0387 mol) of 4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde, 12.4 g (0.193 mol) of hydroxylamine hydrochloride, and 15.8 g (0.193 mol) of sodium acetate were dissolved in 150 mL of EtOH and stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 10) to give 9.6 g of the title compound as a white solid, yield: 73.2%.

[0314] LC-MS: m / z 341.11 [M+H] + .

[0315] Step 5: Preparation of 3-(4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (7e)

[0316] At room temperature, (E)-4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde oxime (5.50 g, 0.0161 mol) and benzyl 3-cyclopropynate (3.88 g, 0.0194 mol) were dissolved in DMF (100 mL), and NaHCO3 (2.70 g, 0.0322 mol) and NCS (2.78 g, 0.0209 mol) were added sequentially. The mixture was stirred overnight at room temperature, quenched with 100 mL of ice water, extracted three times with EA, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 5) to give 7.2 g of the title compound as a yellow solid, yield: 83.7%.

[0317] LC-MS: m / z 539.18 [M+H] + .

[0318] Step 6: Preparation of 3-(4-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (7f)

[0319] At room temperature, 7.20 g (0.0133 mol) of 3-(4-chloro-6-methyl-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester was dissolved in DCM (100 mL), and TFA (12.3 mL) was added. The mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 5) to give 4.2 g of the title compound as a yellow solid, yield: 77.8%.

[0320] LC-MS: m / z 409.10 [M+H] + .

[0321] Step 7: Preparation of 3-(4-chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (7g)

[0322] At room temperature, 3-(4-chloro-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (1.50 g, 0.00367 mol), cyclopentanol (0.632 g, 0.00735 mol), and triphenylphosphine (0.632 g, 0.00550 mol) were dissolved in THF (20 mL). Diisopropyl azodicarbonate (DIAD) (1.11 g, 0.00550 mol) was added at 0 °C, and the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 5) to give 1.4 g of the title compound as a white solid, yield: 77.8%.

[0323] LC-MS: m / z 477.16 [M+H] + .

[0324] Step 8: Preparation of 3-(6-(bromomethyl)-4-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (7h)

[0325] At room temperature, 3-(4-chloro-7-cyclopentyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (1.40 g, 0.00294 mol), NBS (0.630 g, 0.00352 mol), and AIBN (96 mg, 0.000558 mol) were dissolved in DCE (20 mL), heated to 50 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: EA / PE = 1 / 5) to give 1.43 g of the title compound as a pale yellow solid, yield: 88.2%.

[0326] LC-MS: m / z 555.07 [M+H] + .

[0327] Step 9: Preparation of 11-amino-7-cyclopentyl-3-cyclopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (7i)

[0328] At room temperature, 1.43 g (0.00258 mol) of 3-(6-(bromomethyl)-4-chloro-7-cyclopentyl-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester was dissolved in dioxane (20 mL), and ammonia (20 mL) was added. The mixture was heated to 90 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 10 / 1) to give 600 mg of the title compound as a brown solid, yield: 77.8%.

[0329] LC-MS: m / z 365.16 [M+H] + .

[0330] Step 10: Preparation of 11-amino-7-cyclopentyl-3-cyclopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-thione (7g)

[0331] At room temperature, 11-amino-7-cyclopentyl-3-cyclopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (600 mg, 1.64 mmol) was dissolved in 20 mL of ACN, followed by the addition of triethylamine (249 mg, 2.47 mmol) and P2S5 (471 mg, 2.47 mmol). The mixture was heated to 80 °C and stirred overnight. The reaction solution was quenched with 20 mL of ice water, filtered, and the filter cake was washed twice with methanol. The filter cake was dried to give 300 mg of the title compound as a gray solid, yield: 48.1%.

[0332] LC-MS: m / z 381.14 [M+H] + .

[0333] Step 11: Preparation of 7-cyclopentyl-3-cyclopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (7k)

[0334] At room temperature, 11-amino-7-cyclopentyl-3-cyclopropyl-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-thione (300 mg, 0.789 mmol) was dissolved in DCM (5 mL). Trimethyloxonium tetrafluoroboric acid (233 mg, 1.57 mmol) was added at 0 °C, and the mixture was heated to room temperature and stirred overnight. The reaction solution was quenched with 10 mL of ammonium chloride solution, extracted with DCM (20 mL x 2), washed twice with saturated NaCl (20 mL x 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 10 / 1) to give 260 mg of the title compound as a brown solid, yield: 83.6%.

[0335] LC-MS: m / z 395.158 [M+H] + .

[0336] Step 12: 7-Cyclopentyl-3-Cyclopropyl-N 4 Preparation of -(2,2-dimethoxyethyl)-6,7-dihydroisoxazo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4,11-diamine (7l)

[0337] At room temperature, 80.0 mg (0.202 mmol) of 7-cyclopentyl-3-cyclopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine was dissolved in 5 mL of n-butanol, and 106 mg (1.01 mmol) of 2,2-dimethoxyethane-1-amine was added. The mixture was heated to 100 °C and stirred overnight. The reaction solution was concentrated under reduced pressure to give 80 mg of the title compound as a black solid. This crude product was used directly in the next step.

[0338] LC-MS: m / z 452.23 [M+H] + .

[0339] Step 13: Preparation of 9-cyclopentyl-3-cyclopropyl-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (7)

[0340] At room temperature, 7-cyclopentyl-3-cyclopropyl-N 4-(2,2-Dimethoxyethyl)-6,7-dihydroisoxazo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4,11-diamine (80.0 mg, 0.177 mmol) was dissolved in 5 mL of glacial acetic acid and heated to 100 °C with stirring for 2 days. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.1% FA), gradient: 10%-40%) to give 8 mg of the title compound as a white solid, yield: 12.8%.

[0341] LC-MS: m / z 388.18 [M+H] + .

[0342] 1 H NMR (400MHz, DMSO-d6) δ8.15(s,1H),7.53(d,J=1.2Hz,1H),7.46(s,1H),7.08(d,J=1.2Hz,1H),5.55(s,2H),5.28(p,J=8.4H z,1H),3.00(tt,J=7.6,5.7Hz,1H),2.32(q,J=10.2,9.1Hz,2H),2.10–2.02(m,4H),1.73(s,2H),1.24(dd,J=4.9,2.1Hz,4H).

[0343] Example 8: Preparation of 9-cyclopentyl-3-cyclopropyl-6-methyl-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (8)

[0344]

[0345] At room temperature, 7-cyclopentyl-3-cyclopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (7k) (30.0 mg, 0.076 mmol) was dissolved in 1 mL of EtOH, and acetylhydrazine (11.2 mg, 0.152 mmol) was added. The mixture was heated to 90 °C and stirred for 2 days. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.1% FA), gradient: 10%-40%) to give 1.8 mg of the title compound as a white solid, yield: 12.8%.

[0346] LC-MS: m / z 403.19 [M+H] + .

[0347] 1H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.45(s,2H),5.45(s,2H),5.29(p,J=8.7Hz,1H),2.93(tt,J=8.0,5.4Hz ,1H),2.57(s,3H),2.31(q,J=10.5,9.1Hz,2H),2.12–2.00(m,4H),1.74(s,2H),1.30(tt,J=7.3,2.8Hz,4H).

[0348] Example 9: Preparation of 3-cyclopropyl-9-(1-methylcyclopropyl)-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (9)

[0349]

[0350]

[0351] Step 1: Preparation of 4-chloro-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine (9a)

[0352] At room temperature, 2-(4,6-dichloropyrimidin-5-yl)acetaldehyde (5.00 g, 26.3 mmol) and 1-methylcyclopropylamine hydrochloride (3.09 g, 28.9 mmol) were dissolved in 2-methoxy-ethanol (50 mL), and DIEA (16.9 g, 131 mmol) was added. The mixture was heated to 70 °C and stirred overnight in a sealed tube. The reaction solution was quenched with water (30 mL), extracted with DCM (50 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 5:1) to give 4.0 g of the title compound as a pale yellow solid, yield: 73.44%.

[0353] LC-MS: m / z 208.10 [M+H] + .

[0354] Step 2: Preparation of 4-chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine (9b)

[0355] 4-Chloro-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine (3.00 g, 14.5 mmol) was dissolved in THF (30 mL), and LDA (11.6 g, 23.1 mmol) was slowly added dropwise at -78 °C. The mixture was stirred for 30 min, and iodomethane (8.23 g, 58.0 mmol) was added. The mixture was stirred at -78 °C for another 30 min, then brought to room temperature and stirred for 1 h. The mixture was quenched with saturated ammonium chloride, extracted with EA (50 mL x 3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: PE / EA = 5:1) to give 2.7 g of the title compound as a pale yellow solid, yield: 84.31%.

[0356] LC-MS: m / z 221.10 [M+H] + .

[0357] Step 3: Preparation of 4-chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde (9c)

[0358] 4-Chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine (7.90 g, 38.1 mmol) was dissolved in DMF (70 mL), and phosphorus oxychloride (29.1 g, 191 mmol) was added. The mixture was stirred at 70 °C for 5 hours, cooled to room temperature, poured into ice water, and adjusted to alkalinity with saturated NaHCO3. The solution was extracted with EA (30 mL x 3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 8.0 g of the title compound as a pale yellow solid, yield: 84.32%.

[0359] LC-MS: m / z 250.10 [M+H] + .

[0360] Step 4: Preparation of (Z)-4-chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde oxime (9d)

[0361] 4-Chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde (8.00 g, 32.1 mmol) was dissolved in ethanol (80 ml), and sodium acetate (6.58 g, 80.3 mmol) and hydroxylamine hydrochloride (5.58 g, 80.3 mmol) were added. The mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, water was added, and the mixture was stirred for 30 minutes. The mixture was filtered, the filter cake was collected, and dried to obtain 7.4 g of crude product, which was used directly in the next step.

[0362] LC-MS: m / z 265.10 [M+H] + .

[0363] Step 5: Preparation of 3-(4-chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (9e)

[0364] (Z)-4-chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxaldehyde oxime (8.00 g, 30.3 mmol) was dissolved in DMF (80 mL), and benzyl 3-cyclopropynate (7.27 g, 36.3 mmol), sodium bicarbonate (5.21 g, 60.6 mmol), and NCS (5.25 g, 39.3 mmol) were added. The mixture was stirred overnight at room temperature, quenched with water (100 mL), extracted with EA (80 mL x 3), washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 1:1) to give 13.0 g of the title compound as a pale yellow solid, yield: 92.8%.

[0365] LC-MS: m / z 463.10 [M+H] + .

[0366] Step 6: Preparation of 3-(6-(bromomethyl)-4-chloro-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester (9f)

[0367] 12.8 g (27.7 mmol) of 3-(4-chloro-6-methyl-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester was dissolved in DCE (120 mL), and NBS (5.40 g, 30.4 mmol) and AIBN (454 mg, 2.77 mmol) were added. The mixture was stirred overnight at room temperature, and the reaction solution was quenched with water. The mixture was extracted with DCM (30 mL x 3), washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (mobile phase: PE / EA = 5:1) to give 13.0 g of the title compound as a pale yellow solid, yield: 86.9%.

[0368] LC-MS: m / z 541.10 [M+H] + .

[0369] Step 7: Preparation of 11-amino-3-cyclopropyl-7-(1-methylcyclopropyl)-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (9g)

[0370] 13.0 g (24.0 mmol) of 3-(6-(bromomethyl)-4-chloro-7-(1-methylcyclopropyl)-7H-pyrrolo[2,3-d]pyrimidin-5-yl)-5-cyclopropylisoxazole-4-carboxylic acid benzyl ester was dissolved in dioxane (100 mL), and ammonia (100 mL) was added. The mixture was heated to 90 °C and stirred overnight in a sealed tube. The reaction solution was concentrated under reduced pressure, filtered, and the filter cake was collected to give 4.5 g of the title compound as a pale yellow solid. Yield: 53.5%.

[0371] LC-MS: m / z 351.10 [M+H] + .

[0372] Step 8: Preparation of 11-amino-3-cyclopropyl-7-(1-methylcyclopropyl)-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-thione (9h)

[0373] 11-Amino-3-cyclopropyl-7-(1-methylcyclopropyl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-one (3.80 g, 10.4 mmol) was dissolved in acetonitrile (60 ml), and P2S5 (3.11 g, 16.3 mmol) and triethylamine (2.19 g, 21.7 mmol) were added. The mixture was stirred at 80 °C for three days. The reaction solution was cooled to room temperature and slowly added to water (200 ml). The mixture was filtered, and the filter cake was collected to give 7.0 g of the title compound as a brown solid.

[0374] LC-MS: m / z 367.10 [M+H] + .

[0375] Step 9: Preparation of 3-cyclopropyl-7-(1-methylcyclopropyl)-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (9i)

[0376] 11-Amino-3-cyclopropyl-7-(1-methylcyclopropyl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4(5H)-thione (2.00 g, 5.46 mmol) was dissolved in DCM (20 ml), and trimethyloxonium tetrafluoroboric acid (1.60 g, 10.9 mmol) was added at 0 °C. The mixture was stirred at room temperature for 1 hour, and the reaction solution was quenched with saturated sodium bicarbonate. The mixture was extracted with DCM / MeOH = 10:1, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (mobile phase: DCM / MeOH = 10:1) to give 900 mg of the title compound as a pale yellow solid, yield: 43.37%.

[0377] LC-MS: m / z 381.10 [M+H] + .

[0378] Step 10: 3-Cyclopropyl-N 4 Preparation of -(2,2-dimethoxyethyl)-7-(1-methylcyclopropyl)-6,7-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4,11-diamine (9j)

[0379] 100 mg (0.26 mmol) of 3-cyclopropyl-7-(1-methylcyclopropyl)-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine was dissolved in n-butanol (3 ml), and 165 mg (1.57 mmol) of 2,2-dimethoxyethyl-1-amine was added. The mixture was heated to 120 °C and stirred for 48 h in a sealed tube. The reaction solution was then concentrated under reduced pressure to obtain 100 mg of crude product, which was used directly in the next step.

[0380] LC-MS: m / z 438.10 [M+H] + .

[0381] Step 11: Preparation of 3-cyclopropyl-9-(1-methylcyclopropyl)-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (9)

[0382] 3-Cyclopropyl-N 4-(2,2-Dimethoxyethyl)-7-(1-methylcyclopropyl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4,11-diamine (100 mg, 0.228 mmol) was dissolved in AcOH (5 ml), heated to 100 °C and stirred for 48 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%), yielding 28 mg of the title compound as a pale yellow solid, yield: 32.9%.

[0383] LC-MS: m / z 373.10 [M+H] + .

[0384] 1 H NMR(400MHz, DMSO-d6)δ8.18(d,J=8.3Hz,1H),7.60(d,J=1.2Hz,1H),7.46–7.40(m,1H),7.1 0(d,J=1.1Hz,1H),5.65(s,2H),3.02(tt,J=8.0,5.5Hz,1H),1.53(s,3H),1.31–1.18(m,6H).

[0385] Example 10: Preparation of 3-cyclopropyl-6-methyl-9-(1-methylcyclopropyl)-8,9-dihydroimidazo[1,2-a]isoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-13-amine (10)

[0386]

[0387] 3-Cyclopropyl-7-(1-methylcyclopropyl)-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (9i) (100 mg, 0.263 mmol) was dissolved in 2 ml of NMP, and propargylamine (72.3 mg, 0.131 mmol) and p-toluenesulfonic acid (4.50 mg, 0.0260 mmol) were added. The mixture was heated to 180 °C and stirred for 4 h. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water (0.1% FA), gradient: 20%-28%) to obtain 32.0 mg of the title compound as a white solid, yield: 31.4%.

[0388] LC-MS: m / z 388.10 [M+H] + .

[0389] 1 H NMR (400MHz, DMSO-d6) δ8.20 (s, 1H), 7.47 (s, 2H), 6.89 (d, J = 1.2Hz, 1H), 5.44 (s, 2H) ,3.04(tt,J=8.1,5.5Hz,1H),2.42(d,J=1.0Hz,3H),1.53(s,3H),1.32-1.14(m,8H).

[0390] Example 11: Preparation of 3-cyclopropyl-6-methyl-9-(1-methylcyclopropyl)-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (11)

[0391]

[0392] 3-Cyclopropyl-7-(1-methylcyclopropyl)-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidine[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (9i) (80.0 mg, 0.263 mmol) was dissolved in ethanol (3 ml), and acetylhydrazine (38.9 mg, 0.526 mmol) was added. The mixture was heated to 80 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 27.0 mg of the title compound as a pale yellow solid, yield: 26.4%.

[0393] LC-MS: m / z 389.10 [M+H] + .

[0394] 1 H NMR (400MHz, DMSO-d6) δ8.21(s,1H),7.47(s,2H),3.32(s,2H),2.99(ddd,J=13.2,8.2,5.3Hz,1H),2.61(s,3H),1.53(s,3H),1.36-1.21(m,8H).

[0395] Example 12: Preparation of 3-cyclopropyl-9-(1,3-difluoropropyl-2-yl)-8,9-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]tetrazo[1,5-a]azacycloheptane-13-amine (12)

[0396]

[0397] At room temperature, 100 mg (0.256 mmol) of 11-amino-3-cyclopropyl-7-(1,3-difluoroprop-2-yl)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-4-(5H)-thione (59.0 mg, 0.512 mmol) and FeCl3 (83.0 mg, 0.512 mmol) were dissolved in 5 mL of DCM, and TMSN3 (59.0 mg, 0.512 mmol) and FeCl3 (83.0 mg, 0.512 mmol) were added. The mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column: Daisogei 30 mm * 250 mm, C18, 10 μm 100A; mobile phase: acetonitrile / water (0.1% FA); gradient: 10%-40%) to give 6.5 mg of the title compound as a white solid, yield: 6.3%.

[0398] LC-MS: m / z 400.14 [M+H] + .

[0399] 1 H NMR(400MHz,DMSO-d6)δ8.19(s,1H),7.64(s,2H),6.21(s,2H),5.28-5.18(m,1H),5.17- 5.07(m,2H),5.01(dd,J=9.7,4.9Hz,1H),2.92(tt,J=8.1,5.1Hz,1H),1.44-1.33(m,4H).

[0400] Example 13: Preparation of 3-cyclopropyl-9-isopropyl-6-propyl-8,9-dihydroisooxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (13)

[0401]

[0402]

[0403] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (150 mg, 0.407 mmol) was dissolved in ethanol (4 ml), and butyrylhydrazine (83.0 mg, 0.815 mmol) was added. The mixture was heated to 80 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 8 mg of the title compound as a pale yellow solid, yield: 4.8%.

[0404] LC-MS: m / z 404.10 [M+H] + .

[0405] 1 H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.44(s,2H),5.46(s,2H),5.15(p,J=6.8Hz,1H),3.00-2.88(m, 3H), 1.75 (p, J = 7.4Hz, 2H), 1.68 (d, J = 6.7Hz, 6H), 1.30 (tt, J = 8.0, 2.8Hz, 4H), 1.01 (t, J = 7.4Hz, 3H).

[0406] Example 14: Preparation of 6-butyl-3-cyclopropyl-9-isopropyl-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (14)

[0407]

[0408] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (150mg, 0.407mmol) was dissolved in ethanol (4ml), and valerate (90.0mg, 0.815mmol) was added. The mixture was heated to 80°C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 13mg of the title compound as a pale yellow solid, yield: 7.6%.

[0409] LC-MS: m / z 418.10 [M+H] + .

[0410] 1 H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.44(s,2H),5.46(s,2H),5.14(p,J=6.7Hz,1H),3.00-2.88(m ,3H),1.69(d,J=6.8Hz,8H),1.49-1.38(m,2H),1.30(tt,J=8.0,2.8Hz,4H),0.93(t,J=7.4Hz,3H).

[0411] Example 15: Preparation of 3-cyclopropyl-9-isopropyl-6-(trifluoromethyl)-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (15)

[0412]

[0413] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (80.0 mg, 0.217 mmol) was dissolved in ethanol (3 ml), and 2,2,2-trifluoroacetylhydrazine (55.6 mg, 0.434 mmol) was added. The mixture was heated to 80 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by preparative high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 6 mg of the title compound as a pale yellow solid, yield: 6.4%.

[0414] LC-MS: m / z 430.10 [M+H] + .

[0415] 1 H NMR (400MHz, DMSO-d6) δ8.19(s,1H),7.41(s,2H),5.65(s,2H),4.94(p,J=6.9Hz,1 H), 2.93 (tt, J = 8.1, 5.3Hz, 1H), 1.67 (d, J = 6.7Hz, 6H), 1.36 (tt, J = 8.2, 2.9Hz, 4H).

[0416] Example 16: Preparation of 3-cyclopropyl-9-isopropyl-6-(methoxymethyl)-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (16)

[0417]

[0418]

[0419] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (80.0 mg, 0.217 mmol) was dissolved in ethanol (3 ml), and 2-methoxyacetylhydrazine (45.2 mg, 0.434 mmol) was added. The mixture was heated to 80 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 16 mg of the title compound as a pale yellow solid, yield: 18.16%.

[0420] LC-MS: m / z 406.10 [M+H] + .

[0421] 1 H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.44(s,2H),5.56(s,2H),5.06(q,J=6.9Hz,1H),4.81 (s, 2H), 3.36 (s, 3H), 2.94 (tt, J = 8.0, 5.4Hz, 1H), 1.68 (d, J = 6.7Hz, 6H), 1.38-1.24 (m, 4H).

[0422] Example 17: Preparation of 3-cyclopropyl-9-isopropyl-6-(1-methylcyclopropyl)-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (17)

[0423]

[0424] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (80.0 mg, 0.217 mmol) was dissolved in ethanol (3 ml), and 1-methylcyclopropane-1-carbonylhydrazine (49.5 mg, 0.434 mmol) was added. The mixture was heated to 80 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 21.0 mg of the title compound as a pale yellow solid, yield: 23.26%.

[0425] LC-MS: m / z 416.10 [M+H] + .

[0426] 1 H NMR (400MHz, DMSO-d6) δ8.18(s,1H),7.46(s,2H),5.56(s,2H),5.02(q,J=6. 8Hz, 1H), 2.95 (ddd, J=13.5, 8.2, 5.3Hz, 1H), 1.73 (d, J=6.6Hz, 6H), 1.46 (s, 3H), 1.32 (d, J = 2.8Hz, 1H), 1.28 (dd, J = 5.2, 3.1Hz, 2H), 1.24 (s, 1H), 1.06-0.94 (m, 4H).

[0427] Example 18: Preparation of 3-cyclopropyl-9-isopropyl-6-hydroxymethyl-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (18)

[0428]

[0429] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (80.0 mg, 0.217 mmol) was dissolved in ethanol (3 ml), and 2-hydroxyacetylhydrazine (39.1 mg, 0.434 mmol) was added. The mixture was heated to 80 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 7.0 mg of the title compound as a pale yellow solid, yield: 8.2%.

[0430] LC-MS: m / z 416.10 [M+H] + .

[0431] 1 H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.43(s,2H),5.88(t,J=5.6Hz,1H),5.58(s,2H),5.09(p,J=6.8Hz,1H), 4.83(d,J=5.2Hz,2H), 2.93(tt,J=7.9,5.4Hz,1H), 1.67(d,J=6.8Hz,6H), 1.31(ddt,J=8.1,4.4,2.7Hz,4H).

[0432] Example 19: Preparation of 3-cyclopropyl-6,9-diisopropyl-8,9-dihydroisooxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (19)

[0433]

[0434] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (80.0 mg, 0.217 mmol) was dissolved in ethanol (3 ml), and isobutyrylhydrazine (44.3 mg, 0.434 mmol) was added. The mixture was heated to 80 °C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 15.0 mg of the title compound as a pale yellow solid, yield: 17.1%.

[0435] LC-MS: m / z 404.10 [M+H] + .

[0436] 1 H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.43(s,2H),5.46(s,2H),5.15(p,J=6.5Hz,1H),3.49(hept,J=6. 7Hz, 1H), 2.94 (tt, J = 8.1, 5.4Hz, 1H), 1.68 (d, J = 6.7Hz, 6H), 1.34 (d, J = 6.8Hz, 6H), 1.31-1.21 (m, 4H).

[0437] Example 20: Preparation of 3,6-dicyclopropyl-9-isopropyl-8,9-dihydroisooxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-13-amine (20)

[0438]

[0439] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (80mg, 0.217mmol) was dissolved in ethanol (3ml), and cyclopropanecarbazide (43.3mg, 0.434mmol) was added. The mixture was heated to 80°C and stirred overnight. The reaction solution was concentrated under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 21.0mg of the title compound as a pale yellow solid, yield: 24.1%.

[0440] LC-MS: m / z 402.10 [M+H] + .

[0441] 1 H NMR (400MHz, DMSO-d6) δ8.17(s,1H),7.43(s,2H),5.60(s,2H),5.22(t,J=6.8Hz,1H),2.89(tt,J=8.0,5.4Hz,1H), 2.39-2.30(m,1H),1.69(d,J=6.8Hz,6H),1.29(ddt,J=8.1,4.5,2.8Hz,4H),1.13-1.03(m,2H),1.03-0.92(m,2H).

[0442] Example 21: Preparation of 2-(13-amino-3-cyclopropyl-9-isopropyl-8,9-dihydroisoxazolo[4,3-c]pyrimido[5',4':4,5]pyrrolo[3,2-e][1,2,4]triazolo[4,3-a]azacycloheptane-6-yl)acetonitrile (21)

[0443]

[0444] 3-Cyclopropyl-7-isopropyl-4-(methylthio)-6,7-dihydroisoxazolo[4,3-c]pyrimidino[5',4':4,5]pyrrolo[3,2-e]azacycloheptane-11-amine (1k) (80.0 mg, 0.217 mmol) was dissolved in ethanol (3 ml), and 2-cyanoacetylhydrazine (43.0 mg, 0.434 mmol) was added. The mixture was stirred overnight at 80 °C, concentrated directly under reduced pressure, and the residue was purified by high-performance liquid chromatography (HPLC) (column type: Daisogei 30 mm * 250 mm, C18, 10 μm 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to obtain 12.0 mg of the title compound as a pale yellow solid, yield: 13.7%.

[0445] LC-MS: m / z 401.10 [M+H] + .

[0446] 1 H NMR(400MHz,DMSO-d6)δ8.17(s,1H),7.46(s,2H),5.50(s,2H),5.13(p,J=6.8Hz,1H) ,4.65(s,2H),2.95(tt,J=8.1,5.3Hz,1H),1.69(d,J=6.7Hz,6H),1.38–1.26(m,4H).

[0447] Biological evaluation

[0448] Experimental Example 1: Determination of the inhibitory effect of the compound of the present invention on RET kinase activity in vitro

[0449] Experimental materials: HTRF KinEASE-TK kit (Cisbio, 62TK0PEC), RET wild-type (RET WT, Invitrogen, PV3082), RET-V804M kinase (signalchem, R02-12GG-10), RET-G810R kinase (ProQinase, 1724-0000-1), ATP (sigma, A7699), Echo (Labcyte, 550).

[0450] Sample preparation: All compounds were prepared in DMSO, starting at 500 μM, with 3-fold dilutions in 10 gradients. Positive compound LOXO-292 (Selleck, S8781) was prepared in DMSO, starting at 500 μM, with 3-fold dilutions in 10 gradients.

[0451] Prepare 1x kinase reaction buffer: 1 volume of 5X kinase reaction buffer and 4 volumes of water; 5 mM MgCl2; 1 mM DTT.

[0452] Experimental Methods: Using an Echo 550, transfer 50 nmol of the diluted compound to each well of a reaction plate (784075, Greiner). Seal the plate with sealing film and centrifuge at 1000 g for 1 minute. Prepare 2×RET WT using 1× kinase reaction buffer. Add 5 μL of kinase (0.04 ng / μL) to each well of the reaction plate. Seal the plate with sealing film and centrifuge at 1000 g for 30 seconds. Incubate at room temperature for 10 minutes. Prepare a 2× TK-substrate-biotin and ATP mixture using 1× kinase reaction buffer. Add 5 μL of the TK-substrate-biotin (1 μM) / ATP (10 μM) mixture to the reaction plate. Seal the plate with sealing film and centrifuge at 1000 g for 30 seconds. Incubate at room temperature for 30 minutes. Prepare a 2X Sa-XL 665 and TK-antibody-Cryptate mixture using HTRF detection buffer (Cisbio, 62TK0PEC). Add 10 μl of the Sa-XL 665 (62.5 nM) and TK-antibody-Cryptate mixture to each well, centrifuge at 1000g for 30 seconds, and incubate at room temperature for 1 hour. Read the fluorescence signals at 615 nm (Cryptate) and 665 nm (XL665) using an Envision 2104.

[0453] The IC of the compound is obtained using the following nonlinear fitting formula. 50 (Half-maximal inhibitory concentration):

[0454] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope));

[0455] X: Log value of compound concentration;

[0456] Y: Emission Ratio;

[0457] Bottom: minimum value, Top: maximum value, HillSlope: slope;

[0458] The inhibitory activities of the compounds of this invention against RET WT (RET wild-type kinase) are shown in Table 1 below:

[0459] In Table 1, A refers to the inhibitory activity IC of the compound against RET WT. 50 <1nM; B refers to 1nM <IC 50 <10nM; C refers to 10nM <IC 50 <50nM; D refers to IC 50 >50nM.

[0460] Table 1. Inhibitory activity (IC50) of the compounds of this invention against RET WT 50 value

[0461]

[0462] Conclusion: The compounds of this invention can effectively inhibit the activity of both wild-type (RET WT) and mutant RET kinases.

[0463] Experimental Example 2: Cell Proliferation Inhibition Level

[0464] The inhibitory levels of the test compounds on BaF3 KIF5B-RET-WT, BaF3 KIF5B-RET-V804M, and BaF3 KIF5B-RET-G810R stable cell lines (constructed in our own laboratory) were detected, and the results were analyzed based on the IC50 index. 50 Screening for candidate compounds.

[0465] Construction of stable cell lines: Mouse B cells BaF3 (National Experimental Cell Resource Sharing Service Platform, 1101MOU-PUMC000095). The KIF5B-RET WT gene and mutant sequences (G810R, V804M) were synthesized by Suzhou Genewiz Biotechnology Co., Ltd., and cloned into the pMSCV-puromycin vector. Electroporation (Lonza electroporator, AAF-1002B) was used to generate cells expressing the RET fusion mutant gene: BaF3 KIF5B-RET WT, BaF3 KIF5B-RET G810R, and BaF3 KIF5B-RET V804M. Specifically, cells were first cultured in RPMI complete medium (RPMI + 10% FBS + 8 μg / mL puromycin) containing IL3 for 2 weeks to obtain stable cells. These cells were then cultured in selection medium (RPMI complete medium without IL3) for 4 weeks, and stable positive monoclonal cell lines were selected as stable cell lines.

[0466] The specific steps are as follows: Parental BaF3 cells and stable transgenic cell lines were cultured in RPMI 1640 (Invitrogen, A10491-01) with 10% FBS (Gbico, 10099141), penicillin and streptomycin (1% penicillin and streptomycin, Gibco, 15140122), and Zeocin (1000 μg / mL, Invitrogen, R25001). Cells in the logarithmic growth phase were collected, and cell viability was assessed using the trypan blue method to ensure greater than 90%. Cells were seeded in 384-well plates (Corning, 3570) with a white transparent bottom at 450 cells / well, and the experimentally tested compounds were added.

[0467] The compounds were dissolved and diluted in DMSO; starting at a concentration of 10 mM, 3-fold dilutions were performed to establish 10 concentration gradients, with 3 replicates per gradient. Cells were co-cultured at 37°C and 5% CO2 for 72 h. Cell proliferation levels were determined by detecting total ATP content using a CELL Titer-GLO luminescence assay.

[0468] Cells were removed from 384-well plates and equilibrated at room temperature for 30 min. 20 μL of CellTiter Glo (Promega, catalog number G7572) was added to each well, vortexed, and incubated at room temperature for 10 min. Luminescence values ​​were read using a Biotek Cytation 3 microplate reader. Experimental data were plotted with the compound concentration (Logarithm) on the X-axis and the percentage inhibition level on the Y-axis. A nonlinear fit was performed using GraphPad Prism 8.0 software to determine the dose-effect relationship. The IC50 of the compound on cell proliferation inhibition was calculated using the following formula. 50 value:

[0469] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0470] X represents the log value of the compound concentration; Y represents the level of inhibition against the kinase; Top and Bottom represent the Y values ​​at the highest and lowest plateau phases of the curve; Hillslope is the Hill constant.

[0471] The inhibitory activity of the compounds of this invention on cell proliferation is shown in Table 2 below:

[0472] In Table 2, A refers to the inhibitory activity (IC50) of the compound on cell proliferation. 50 <10nM; B refers to 10nM <IC 50 <100nM; C refers to 100nM <IC 50 <500nM; D refers to IC 50 >500nM.

[0473] Table 2. Inhibitory effect of the compounds of the present invention on BaF3 KIF5B-RET-WT, BaF3 KIF5B-RET-V804M, and BaF3 KIF5B-RET-G810R cells. 50 value

[0474]

[0475]

[0476] Conclusion: The compounds of this invention can effectively inhibit the cell proliferation of RET WT and RET-resistant mutants.

[0477] Experimental Example 3: Pharmacokinetic Evaluation of the Compound ICR in Mice

[0478] Male 7-8 week old ICR mice (Beijing Vital River Laboratory Animal Technology Co., Ltd.) were housed in an SPF environment at a temperature of 20-26°C, with a daily temperature difference not exceeding 4°C and a relative humidity of 40-70% RH, with alternating 12h / 12h lighting daily. After a 3-5 day acclimatization period, animals receiving oral administration were fasted overnight (>12h) one day before the experiment, but water was allowed. The compound of this invention was administered orally (weighed 2.5mg, added to a 100% mixed solvent (10% NMP + 49% PEG400 + 1% Tween 80 + 40% water) to a final volume of 5mL, and then sonicated until a homogeneous solution was obtained). Blood samples were collected from the orbital rim before administration and at 15min, 30min, 1h, 2h, 4h, 6h, and 8h after administration. The blood was anticoagulated with dipotassium EDTA, centrifuged at 3500rpm for 10 minutes at 4°C, and plasma was obtained and stored at -20°C until testing.

[0479] Take 50 μL of plasma sample into a 1.5 mL EP tube, add 400 μL of acetonitrile solution containing internal standard, vortex for 1 minute to mix thoroughly, and centrifuge at 10000 rpm for 10 minutes. Transfer 0.2 mL of supernatant, filter through a 0.22 μM organic membrane, and add to a vial. Analyze the compound concentration in the sample using LC-MS / MS. The plasma concentration-time curve of the compound in mice and the main pharmacokinetic parameters (AUC) were calculated using MAS Studio (V1.3.1 stable) software. 0-t C max T max T 1 / 2 And F%, F% = (AUC) po ×Dose iv ) / (AUC iv ×Dose po )×100%.

[0480] Pharmacokinetic data of the compounds of this invention after oral administration are shown in Table 3:

[0481] In Table 3, A refers to the AUC of the compound. 0-t (μg / L*h)<5000; B refers to 5000 <AUC 0-t (μg / L*h)<10000; C refers to 10000 <AUC 0-t (μg / L*h)<20000; D refers to 20000 <AUC 0-t (μg / L*h)<30000.

[0482] Table 3. Pharmacokinetic parameters of the compounds of this invention administered orally to male ICR mice after a single oral administration.

[0483]

[0484]

[0485] Conclusion: The compound of this invention exhibits good in vivo pharmacokinetic properties when orally administered to mice.

[0486] Experimental Example 4: Efficacy Experiment of the Compound of the Invention in Mouse Subcutaneous Xenograft Tumors

[0487] The mouse B cell line, BaF3, was derived from the Peking Union Medical College Hospital Cell Resource Center. Vector construction was commissioned to Suzhou Genewiz Biotechnology Co., Ltd., and our laboratory obtained BaF3 KIF5B RET-G810R cells through electroporation and screening for positive clones.

[0488] Female Balb / c Nude mice aged 7-8 weeks were used as experimental animals, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. They were housed in an SPF environment with a temperature of 20-26℃, a daily temperature difference not exceeding 4℃, and a relative humidity of 40-70% RH, with alternating 12h / 12h lighting daily. The animals underwent an acclimatization period of 3-5 days.

[0489] Two weeks prior to animal inoculation, target cells were resuscitated and cultured in RPMI-1640 medium (10% FBS, 1% PS, 0.8 μg / mL puro) at 37°C in a 5% CO2 environment to amplify the cells. Cells in the logarithmic growth phase were collected, washed twice with PBS buffer, and counted. The cell suspension was centrifuged, and the supernatant was discarded. A 1:1 mixture of matrix gel and PBS buffer was added to the cell pellet to prepare a concentration of 5 × 10⁻⁶ cells / mL. 7 Prepare a 1 mL / ml cell suspension and place it on ice. Wipe the mouse inoculation site with an alcohol swab, and subcutaneously inoculate cells into the right forelimb of the mouse using a pre-cooled 1 mL syringe. The inoculation volume is 100 μL (inoculation amount is 5 × 10⁶ cells / mL). 6 / Only).

[0490] The experiment was divided into 10 groups: a solvent control group, and three dosage groups (5 mg / kg, 10 mg / kg, and 20 mg / kg) for Examples 4, 15, and 19, respectively. Administration was by gavage, twice daily. Solvent: 20% HP-β-CD. Mice were weighed three times a week, and tumors were measured three times a week using calipers. The formula V = major axis × minor axis was used. 2 / 2, calculate the tumor volume V; according to the formula TGI(%) = [1 - (average tumor volume at the end of treatment - average tumor volume at the beginning of treatment) / (average tumor volume at the end of treatment in solvent control group - average tumor volume at the beginning of treatment in solvent control group)] × 100%, calculate the tumor growth inhibition rate TGI of the compound.

[0491] The efficacy results of the compounds of this invention against subcutaneous xenograft tumors in mice are as follows: Figures 1A-1CAs shown, the mouse's weight is as follows Figures 2A-2C As shown.

[0492] Conclusion: As can be seen from Figures 1 and 2, the compound of the present invention has a good tumor-suppressing effect on a mouse subcutaneous xenograft model, and the mice showed no abnormal weight gain.

Claims

1. A compound of general formula (III) or a pharmaceutically acceptable salt thereof, (III) in, Z is selected from C or N; Cycle B is selected from 5- to 6-membered heteroaryl groups, which may be further selectively bound by one or more R groups. 6 Replaced; R 1 Selected from C1-C6 alkyl, C1-C6 haloalkyl, and C3-C6 cycloalkyl, wherein the C3-C6 cycloalkyl may optionally be further substituted with C1-C6 alkyl; R 2 and R 3 It is hydrogen; R 5 Selected from C3-C6 cycloalkyl groups; R 6 The group is selected from hydrogen, C1-C6 alkyl, and C3-C6 cycloalkyl, wherein the C1-C6 alkyl and C3-C6 cycloalkyl are optionally further substituted by one or more substituents selected from halogen, cyano, hydroxy, C1-C6 alkyl, and C1-C6 alkoxy.

2. The compound of formula (III) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein ring B is selected from pyridinyl, pyrimidinyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, and isoxazolyl, optionally further divided by one or more R... 6 Replaced; R 6 As defined in claim 1.

3. The compound of formula (III) according to claim 1, or a pharmaceutically acceptable salt thereof, is a compound of formula (IV) or a pharmaceutically acceptable salt thereof. (IV) in, X1 is CH or N; X2 is CH or N; R 1 R 2 R 3 R 5 R 6 As defined in claim 1.

4. The compound of formula (III) according to claim 1, or a pharmaceutically acceptable salt thereof, wherein it is the compound of formula (VI) or a pharmaceutically acceptable salt thereof. (VI) in, X5, X6, and X7 are each independently selected as CH or N; R 1 R 2 R 3 R 5 R 6 As defined in claim 1.

5. The compound of formula (III) according to claim 1, or a pharmaceutically acceptable salt thereof, selected from: , , , , , , , , , , , , , , , , , , , and .

6. A method for preparing a compound of general formula (IV) or a pharmaceutically acceptable salt thereof, comprising the following steps: In the presence of a catalyst, compounds of formula IVj undergo cyclization to give compounds of general formula (IV); Among them, X1, X2, R 1 R 2 R 3 R 5 R 6 As defined in claim 3.

7. A pharmaceutical composition comprising a compound of formula (III) according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

8. Use of the compound of general formula (III) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7, in the preparation of a RET kinase inhibitor.

9. Use of the compound of general formula (III) according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 7, in the preparation of a medicament for the prevention and / or treatment of diseases related to RET kinase activity.

10. The use according to claim 9, wherein the disease is a malignant tumor disease.

11. The use according to claim 10, wherein the malignant tumor disease is non-small cell lung cancer or thyroid cancer.

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