Aromatic ring-fused thiazine derivative and pharmaceutical use thereof

Abstract

The present invention belongs to the technical field of pharmaceuticals, relates to an aromatic ring-fused thiazine derivative, and in particular relates to a free alkali, an isomer, a pharmaceutically acceptable salt form and other structural forms of the compound, a method for preparing such compounds, and a composition comprising such compounds and the therapeutic use thereof. In the present invention, on the basis of the histamine PI3Kδ receptor ligand, a series of compounds with novel structures have been developed, and a series of related biological experiments have been carried out on the compounds. The experimental results all show that the compounds are found to be a novel type of highly selective PI3Kδ inhibitor, which combines the characteristics of low toxicity and mild side effects and low bioavailability, is used for developing inhaled drugs suitable for treating respiratory diseases such as asthma and COPD, and has important clinical significance and social values.

Description

Aromatic ring thiazine derivatives and their medical uses Technical Field

[0001] The present invention belongs to the field of medical technology and relates to an aromatic ring thiazine derivative, specifically to the free base, isomers, and pharmaceutically acceptable salt forms of the compound; a method for preparing the compound; a composition containing the compound and its therapeutic use. Background Art

[0002] The PI3K / Akt / mTOR pathway is a signal transduction pathway that is abnormally activated in human cancers and is closely associated with the development and progression of malignant tumors. As a key kinase in this signaling pathway, PI3K has attracted numerous clinical trials and is a popular target for anti-tumor drug development.

[0003] PI3K signaling is one of the most commonly aberrantly activated pathways. Early studies have shown that the pan-PI3K inhibitors LY294002 and wortmannin can reverse resistance in cancer cells to multiple therapies. Several PI3K family members are also involved in inflammation and autoimmunity. Class I PI3Ks consist of a complex regulatory subunit with a p110 catalytic subunit (p110α, β, γ, or δ). These heterodimeric complexes are termed PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ, with p110α, p110β, p110γ, and p110δ representing the catalytic subunits themselves. While p110α and p110β exhibit widespread tissue distribution, the p110γ and p110δ isoforms are primarily expressed in leukocytes and are important in the activation of immune responses, such as leukocyte migration, B and T cell activation, and mast cell degranulation. Consequently, the PI3Kδ and PI3Kγ isoforms are strongly implicated in inflammatory respiratory diseases.

[0004] Currently, there are only four types of PI3Kδ inhibitors on the market, including Idelalisib, Copanlisib, Duvelisib, and Umbralisib. Idelalisib and Duvelisib have similar structures. The parent nucleus of Idelalisib is quinazolinone, and Duvelisib is modified based on Idelalisib.

[0005] Patent WO2005113556A1, corresponding to its Chinese counterpart CN101031569B, protects the structure of the PI3Kδ inhibitor idelalisib (CAL-101, zydelig). In July 2014, idelalisib received US FDA approval for the treatment of relapsed chronic lymphocytic leukemia (CLL), follicular B-cell non-Hodgkin lymphoma (FL), and small lymphocytic lymphoma (SLL). However, the drug label for idelalisib carries a black box warning regarding possible side effects such as hepatotoxicity, pneumonitis, severe diarrhea, enteritis, and intestinal perforation.

[0006] Patent WO2011008302A1, corresponding to its Chinese counterpart CN102711767B, protects the PI3Kδ / γ selective inhibitor duvelisib (IPI-145, INK-1197). Duvelisib received US FDA approval in September 2018 for the treatment of relapsed or refractory chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL) in patients who have received at least two prior therapies, and for the treatment of relapsed or refractory follicular lymphoma (FL) in adult patients who have received at least two prior therapies. It is important to note that the duvelisib package insert contains a boxed warning regarding life-threatening and serious toxic effects, including infection, diarrhea or colitis, skin reactions, and pneumonitis.

[0007] Based on the toxic and side effects of the above two PI3Kδ inhibitors, domestic and foreign researchers continue to modify the compound structure based on their structure, hoping to find safer and more effective PI3Kδ inhibitors.

[0008] Patent US20130053362A1, corresponding to the Chinese equivalent CN103998442A, discloses a compound and pharmaceutical composition that inhibits phosphoinositide 3-kinase (PI3K). The parent nucleus of the compound is the structure of isoquinolinone and quinazolinone, and can be used to treat cancer, inflammatory diseases or autoimmune diseases.

[0009] Patent CN107033145B protects a class of benzothiazine and benzothiadiazine compounds, most of which selectively inhibit PI3Kδ and can be used to prepare anti-inflammatory and anti-tumor drugs. Among them, the representative compound SI-11 has an IC of 1.177 for PI3Kδ. 50The range is 0-20 nM. It can be seen from the literature (Eur. J. Med. Chem. 2019, 170, 112-125) that the oral bioavailability of this molecule (S-63 in the literature) is 29.2%.

[0010] In summary, the parent nuclei of the above-mentioned PI3K inhibitors are mostly based on isoquinolinone and quinazolinone, and are all used to prepare drugs for anti-tumor, anti-inflammatory, and immune diseases. Patent CN107033145B, based on Idelalisib and Duvelisib, adopts the principle of bioisosteres to design compounds with thiazine and benzothiadiazine as the skeleton, which are also used for the treatment of tumor diseases and are suitable for oral administration. Therefore, in the field of PI3Kδ inhibitors, PI3Kδ has high selectivity and is administered by inhalation. Research on structural types for the treatment of respiratory diseases such as asthma and COPD is still relatively scarce.

[0011] Therefore, those skilled in the art are in urgent need of searching for and discovering novel highly selective PI3Kδ inhibitors with minimal toxic side effects and low bioavailability, which can be used to develop inhaled drugs suitable for the treatment of respiratory diseases such as asthma and COPD, which have important clinical significance and social value. Summary of the Invention

[0012] The present invention aims to provide a novel aromatic ring thiazine derivative, which is a highly selective PI3Kδ inhibitor with zero oral bioavailability and a fast clearance rate. It can be used to develop inhalation preparations for the treatment of respiratory diseases such as asthma and COPD.

[0013] In order to solve the above technical problems, the technical solutions adopted by the present invention are as follows:

[0014] The present invention provides an aromatic ring thiazine derivative, which is a compound represented by general formula I or its isomers, or a pharmaceutically acceptable salt thereof:

[0015] Where:

[0016] W is selected from direct bond, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl or C 1-6 Alkylamine;

[0017] Ring A is selected from a benzene ring or a 5-8 membered aromatic heterocycle substituted by at least one R0, R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 2-6 Alkynyl, C 2-6 Alkenyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C1-6 Alkoxy, C 1-6 One or more of alkylamino, amino, cyano, hydroxyl, carboxyl, carbonyl, and keto groups;

[0018] R1 and R2 are each independently selected from hydrogen, halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 Heterocyclic group, substituted or unsubstituted C 1-6 Alkylamino, cyano, hydroxyl, carboxyl or carbonyl; C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 3-6 Heterocyclic group, C 1-6 Alkylamino groups may be substituted with at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino groups;

[0019] Or, R1 and R2 are connected to form a monocyclic or bicyclic structure;

[0020] R3, R4, and R5 are each independently selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Alkylamino, amino or cyano groups;

[0021] R6 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 2-6 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl, substituted or unsubstituted 5-8 membered heteroaryl, amino, cyano or hydroxy; C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl and 5-8 membered heteroaryl can be replaced by at least one R 11 Replacement, R 11 Selected from or a substituted or unsubstituted 5-8 membered aromatic ring or aromatic heterocycle, wherein the 5-8 membered aromatic ring or aromatic heterocycle may be substituted by at least one of the following groups: hydrogen, halogen or C 1-6 alkyl;

[0022] R7, R8, R9, R 10 Each independently selected from C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0023] n, p, and q are each independently selected from 0, 1, 2, 3, 4, 5, or 6.

[0024] The present invention also provides an aromatic ring thiazine derivative, which can be a compound represented by general formula II or its isomers, or a pharmaceutically acceptable salt thereof:

[0025] Where:

[0026] W is selected from a direct bond or C 1-6 alkyl;

[0027] Ring A is selected from a benzene ring, pyrazole or pyridine substituted by at least one R0, R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 1-6 Alkylamino, amino or cyano groups;

[0028] R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 Heterocyclic or cyano; C 1-6 Alkyl, C 3-6 Cycloalkyl, C 3-6 The heterocyclic groups may be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino groups;

[0029] R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano;

[0030] R6 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted 5-8 membered heteroaryl; C 1-6 Alkyl and 5-8 membered heteroaryl groups may be replaced by at least one R 11 Replacement, R 11 Selected from

[0031] R9 is selected from C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0032] n, p, q are each independently selected from 0, 1, 2 or 3.

[0033] Preferably, in the above general formula II, when ring A is a benzene ring substituted with at least one R0, R1 is selected from C 1-6 Alkyl; or, when ring A is pyrazole or pyridine substituted with at least one R0, R1 is selected from C 1-6 Alkyl or C 3-6 Cycloalkyl.

[0034] Preferably, in the above general formula II, W is selected from C 1-6 alkyl.

[0035] Preferably, in the above general formula II, W is selected from C 1-6 Alkyl, ring A is selected from benzene ring, pyrazole or pyridine, R1 is selected from C 1-6 Alkyl, R3 is selected from cyano, R4 is selected from C 1-6 Alkyl, R5 is selected from amino, and R6 is selected from halogen.

[0036] The present invention also provides an aromatic ring thiazine derivative, which can be a compound represented by general formula II-1 or its isomers, or a pharmaceutically acceptable salt thereof:

[0037] Where:

[0038] Ring A is selected from a benzene ring substituted with at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 1-6 Alkylamino, amino or cyano groups;

[0039] R1 is selected from halogen, substituted or unsubstituted C 2-6 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 Heterocyclic or cyano; C 2-6 Alkyl, C 3-6 Cycloalkyl, C 3-6 The heterocyclic groups may be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino groups;

[0040] R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano;

[0041] R6 is selected from halogen, C 1-6 alkyl or 5-8 membered heteroaryl.

[0042] Preferably, in the aforementioned general formula II-1, ring A is selected from a benzene ring.

[0043] Preferably, in the above general formula II-1, R1 is selected from substituted or unsubstituted C 3-6 Cycloalkyl, when C 3-6 When a cycloalkyl group has a substituent, it may be substituted by at least one of the following groups: halogen, C 1-6 Alkyl, hydroxy or amino.

[0044] Preferably, in the aforementioned general formula II-1, R1 is selected from C3-6 Cycloalkyl.

[0045] Preferably, in the above general formula II-1, R1 is selected from substituted or unsubstituted C 2-6 Alkyl, when C 2-6 When an alkyl group has a substituent, it may be substituted by at least one of the following groups: halogen, hydroxyl or amino.

[0046] Preferably, in the aforementioned general formula II-1, R3 is selected from cyano.

[0047] Preferably, in the aforementioned general formula II-1, R4 is selected from C 1-6 alkyl.

[0048] Preferably, in the aforementioned general formula II-1, R5 is selected from amino group.

[0049] Preferably, in the above general formula II-1, R6 is selected from halogen or C 1-6 alkyl.

[0050] Preferably, in the above general formula II-1, ring A is selected from a benzene ring, and R1 is selected from a C 3-6 Cycloalkyl, R3 is selected from cyano, R4 is selected from C 1-6 Alkyl, R5 is selected from amino, and R6 is selected from halogen.

[0051] The present invention also provides an aromatic ring thiazine derivative, which can be a compound represented by general formula II-2 or its isomers, or a pharmaceutically acceptable salt thereof:

[0052] Where:

[0053] Ring A is selected from pyrazole or pyridine substituted with at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 1-6 Alkylamino, amino or cyano groups;

[0054] R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 Heterocyclic or cyano; C 1-6 Alkyl, C 3-6 Cycloalkyl, C 3-6 The heterocyclic groups may be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino groups;

[0055] R3, R4, and R5 are each independently selected from C 1-6Alkyl, amino or cyano; and R4 and R5 are not amino at the same time;

[0056] R6 is selected from halogen, C 1-6 alkyl or 5-8 membered heteroaryl.

[0057] Preferably, in the above general formula II-2, ring A is selected from pyrazole or pyridine substituted with at least one R0, and R0 is selected from hydrogen, halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl.

[0058] Preferably, in the aforementioned general formula II-2, R1 is selected from substituted or unsubstituted C 1-6 Alkyl, when C 1-6 When an alkyl group has a substituent, it may be substituted by at least one of the following groups: halogen, hydroxyl or amino.

[0059] Preferably, in the aforementioned general formula II-2, R3 is selected from cyano.

[0060] Preferably, in the aforementioned general formula II-2, R4 is selected from C 1-6 alkyl.

[0061] Preferably, in the aforementioned general formula II-2, R5 is selected from amino group.

[0062] Preferably, in the above general formula II-2, R6 is selected from halogen or C 1-6 alkyl.

[0063] Preferably, in the aforementioned general formula II-2, ring A is selected from pyrazole, pyridine or pyridine substituted by at least one R0, R0 is selected from halogen, R1 is selected from C 1-6 Alkyl, R3 is selected from cyano, R4 is selected from C 1-6 Alkyl, R5 is selected from amino, and R6 is selected from halogen.

[0064] The present invention also provides an aromatic ring thiazine derivative, which can be a compound represented by general formula III or its isomers, or a pharmaceutically acceptable salt thereof:

[0065] Where:

[0066] Ring A is selected from a benzene ring, pyrazole or pyridine substituted by at least one R0, R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or halogenated C 1-6 alkoxy;

[0067] R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl or substituted or unsubstituted C 3-6 Cycloalkyl; C 1-6 Alkyl, C3-6 The cycloalkyl groups may be substituted with at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino groups;

[0068] R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano;

[0069] R 11 Selected from or substituted or unsubstituted 5-8 membered aromatic ring or aromatic heterocycle; wherein the 5-8 membered aromatic ring or aromatic heterocycle may be substituted by at least one of the following groups: hydrogen or C 1-6 alkyl;

[0070] R7, R8, R9, R 10 Each independently selected from C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0071] n, p, q are each independently selected from 0, 1, 2 or 3.

[0072] The present invention also provides an aromatic ring thiazine derivative, which can be a compound represented by general formula III or its isomers, or a pharmaceutically acceptable salt thereof:

[0073] Where:

[0074] Ring A is selected from a benzene ring, pyrazole or pyridine substituted by at least one R0, R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or halogenated C 1-6 alkoxy;

[0075] R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl or substituted or unsubstituted C 3-6 Cycloalkyl; C 1-6 Alkyl, C 3-6 The cycloalkyl groups may be substituted with at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino groups;

[0076] R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano;

[0077] R 11 Selected from

[0078] R9, R 10 Each independently selected from substituted C 1-6 Alkyl, substituted or unsubstituted 5-8 membered heterocyclic group, when C1-6 When an alkyl group or a 5-8 membered heterocyclic group has a substituent, it may be substituted by at least one of the following groups: 1-6 alkoxy;

[0079] n is selected from 0, 1, 2 or 3.

[0080] In the above general formula III, ring A is selected from a benzene ring substituted by at least one R0, and R0 is selected from hydrogen, halogen or C 1-6 Alkyl; R1 is selected from C 1-6 Alkyl; R3, R4, R5 are each independently selected from C 1-6 Alkyl, amino or cyano;

[0081] R 11 Selected from

[0082] Preferably, R3, R4, and R5 in the aforementioned general formula I, general formula II, general formula II-1, general formula II-2, and general formula III can be further defined as follows: R3 is selected from cyano, R4 is selected from C 1-6 Alkyl, R5 is selected from amino.

[0083] Preferably, in the aforementioned aromatic thiazine derivatives, the heterocyclic group, heteroaryl group, or aromatic heterocycle contains at least one heteroatom, and the heteroatom is selected from N, O, or S.

[0084] More preferably, the aromatic heterocycle may include, but is not limited to, furan, thiophene, pyrazole, pyridine, pyrimidine, and the like.

[0085] More preferably, the heteroaryl group may include, but is not limited to, a group selected from furyl, thienyl, pyrazolyl, pyridinyl, pyrimidinyl, etc. More preferably, the heterocyclic group may include, but is not limited to, a group selected from pyrrole, morpholine, piperidine, piperazine, etc.

[0086] The present invention also provides an aromatic ring thiazine derivative, including the following compounds numbered BIOS-A1 to BIOS-A8, BIOS-B1 to BIOS-B17 or pharmaceutically acceptable salts thereof:

[0087] BIOS-A1: (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-(1H-pyrazol-4-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0088] BIOS-A2: (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0089] BIOS-A3: (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-(pyridin-4-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0090] BIOS-A4: (S)-2-amino-4-((1-(8-fluoro-1,1-dihydroxy-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0091] BIOS-A5: (S)-2-amino-4-((1-(8-fluoro-2-(5-fluoropyridin-3-yl)-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0092] BIOS-A6: (S)-2-amino-4-((1-(8-chloro-2-(5-fluoropyridin-3-yl)-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0093] BIOS-A7: (S)-2-amino-4-((1-(2-benzyl-8-chloro-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0094] BIOS-A8: (S)-6-(3-(1-((2-amino-5-cyano-6-methylpyrimidin-4-yl)amino)propyl)-2-(5-fluoropyridin-3-yl)-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-8-yl)-N,N-bis(2-methoxyethyl)hex-5-ynamide;

[0095] BIOS-B1: (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0096] BIOS-B2: (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)-3,3-difluoropropyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0097] BIOS-B3: (S)-2-amino-4-(((8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)(cyclobutyl)methyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0098] BIOS-B4: (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)-2-methylpropyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0099] BIOS-B5: (S)-2-amino-4-(((8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)(cyclopropyl)methyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0100] BIOS-B6: (S)-2-amino-4-(6-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)-5-azaspiro[2.4]heptane-5-yl)-6-methylpyrimidine-5-carbonitrile;

[0101] BIOS-B7: (S)-2-amino-4-(2-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)pyrrolidin-1-yl)-6-methylpyrimidine-5-carbonitrile;

[0102] BIOS-B9: (S)-2-amino-4-((1-(8-fluoro-1,1-dihydroxy-2-(pyridin-3-ylmethyl)-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0103] BIOS-B10: (S)-2-amino-4-((1-(8-fluoro-2-((5-fluoropyridin-3-yl)methyl)-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0104] BIOS-B11: (S)-2-amino-4-((1-(8-(3-(2-(2-methoxyethoxy)ethoxy)prop-1-yn-1-yl)-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0105] BIOS-B12: (S)-6-(3-(1-((2-amino-5-cyano-6-methylpyrimidin-4-yl)amino)propyl)-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-8-yl)-N,N-bis(2-methoxyethyl)hex-5-ynamide;

[0106] BIOS-B13: (S)-2-amino-4-((1-(8-(3-(2-(2-methoxyethoxy)ethoxy)prop-1-yn-1-yl)-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0107] BIOS-B14: (S)-2-amino-4-methyl-6-((1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)pyrimidine-5-carbonitrile;

[0108] BIOS-B15: (S)-2-amino-4-methyl-6-((1-(8-(6-morpholino-6-carbonylhex-1-yn-1-yl)-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)pyrimidine-5-carbonitrile;

[0109] BIOS-B16: (S)-2-amino-4-((1-(1,1-dihydroxy-8-(6-carbonyl-6-(pyrrolidin-1-yl)hex-1-yn-1-yl)-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0110] BIOS-B17: (S)-2-amino-4-((1-(8-(2-methoxypyridin-4-yl)-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile;

[0111] The aforementioned compounds numbered BIOS-A1 to BIOS-A8, BIOS-B1 to BIOS-B17, or pharmaceutically acceptable salts thereof, have the following structural formulas:

[0112] The present invention also provides a method for preparing the aforementioned compound, comprising the following steps (including but not limited to the following method):

[0113] Route 1: Preparation of series A compounds

[0114] The specific reaction process is as follows:

[0115] An ortho-halogenated aromatic amine compound A-001 reacts with hydrochloric acid, sodium nitrite, sodium sulfite, and copper sulfate to produce a sulfonyl chloride intermediate A-002; this intermediate then undergoes a condensation reaction with an amino compound to produce an intermediate A-003; chiral (S)-2-(but-3-yn-2-yl)isoindoline-1,3-dione A-004 and intermediate A-003 undergo a coupling ring closure reaction in the presence of a metal catalyst to produce intermediate A-005; intermediate A-005 and hydrazine hydrate are heated to reflux in an ethanol solvent and subjected to a deprotection reaction to produce intermediate A-006, which then undergoes an SN2 reaction with a substituted aminopyrimidine to produce a series of target compounds, including but not limited to compounds BIOS-A1 to BIOS-A7.

[0116] In addition, the product BIOS-A6 prepared by this reaction route can be further obtained into the BIOS-A8 compound through a one-step reaction with an alkaline derivative using a noble metal as a catalyst under alkaline conditions.

[0117] Route 2: Preparation of series B compounds

[0118] The specific reaction process is as follows:

[0119] An ortho-halogenated aromatic amine compound B-001 reacts with hydrochloric acid, sodium nitrite, sodium sulfite, and copper sulfate to produce a sulfonyl chloride intermediate B-002, which then undergoes a condensation reaction with an amino compound to produce intermediate B-003. A Boc-protected chiral amino acid intermediate B-004 reacts with intermediate B-003 at low temperature to produce intermediate B-005. Intermediate B-005 is reacted in concentrated hydrochloric acid to produce B-006, which then undergoes an SN2 reaction with a substituted aminopyrimidine to produce a series of target compounds, including but not limited to compounds BIOS-B1 to BIOS-B10.

[0120] In addition, the products prepared by this reaction route can be further obtained into BIOS-B11 to BIOS-B17 compounds through a one-step reaction with alkaline derivatives using noble metals as catalysts.

[0121] The "compounds" described in the present invention include but are not limited to the following forms of compounds: free base, stereoisomers, geometric isomers, tautomers, isotopes, pharmaceutically acceptable salts, solvates, hydrates, prodrugs (esters or phosphates), etc.

[0122] The "compounds" described herein may be asymmetric, for example, having one or more stereoisomers. Unless otherwise indicated, all stereoisomers include, for example, enantiomers and diastereomers. Compounds of the present invention containing asymmetric carbon atoms may be isolated in optically pure or racemic forms. Optically pure forms may be obtained by resolution of racemic mixtures or by synthesis using chiral starting materials or reagents.

[0123] The “isomers” described in the present invention refer to stereoisomers or tautomers unless otherwise specified. Unless otherwise specified, the term “stereoisomer” refers to compounds with the same chemical structure but different arrangements of atoms or groups in space. Stereoisomers include, but are not limited to, enantiomers, diastereomers, conformers (rotational isomers), geometric isomers (cis / trans) isomers, and atropisomers. Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on the differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization. Unless otherwise specified, the term “tautomer” refers to structural isomers with different energies that can be converted into each other through a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be reached. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations.Valence tautomers include interconversions by reorganization of some of the bonding electrons.

[0124] The term "isotope" as used herein means, unless otherwise specified, that the compounds of the present invention may be present in an isotopically labeled or enriched form, containing one or more atoms whose atomic mass or mass number differs from the atomic mass or mass number of the largest atom found in nature. Isotopes may be radioactive or non-radioactive. Isotopes commonly used as isotope labels are: hydrogen isotopes, including but not limited to: 2 H and 3 H; Carbon isotopes: including but not limited to 13 C and 14 C; Chlorine isotopes: including but not limited to 35 Cl and 37 Cl; Fluorine isotopes: including but not limited to 18 F; Iodine isotopes: including but not limited to 123 I and 125 I; Nitrogen isotopes: including but not limited to 13 N and 15 N; oxygen isotopes: including but not limited to 15 O. 17 O and 18O; Sulfur isotopes: including but not limited to 35 These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues, especially 3 H and 13 C, because they are easy to label and detect, they are more widely used. Some heavy isotopes, such as deuterium ( 2 H) substitution can enhance metabolic stability and prolong half-life, thereby achieving the goal of reducing dosage and providing therapeutic advantages. Isotope-labeled compounds are generally synthesized from labeled starting materials using known synthetic techniques similar to those used for synthesizing non-isotope-labeled compounds.

[0125] The term "pharmaceutically acceptable salt" as used herein refers to a salt of a compound of the present invention, prepared by reacting a compound having a specific substituent discovered herein with a relatively nontoxic base. When the compound of the present invention contains relatively acidic functional groups, a base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, magnesium salts, ammonium, or organic amines. Examples include alkali metal salts, alkaline earth metal salts, other metal salts, inorganic base salts, organic base salts, inorganic acid salts, lower alkane sulfonates, aryl sulfonates, organic acid salts, and amino acid salts.

[0126] In addition to the form of salts, the compounds provided by the present invention also exist in prodrug form. The prodrugs of the compounds described herein easily undergo chemical changes under physiological conditions to be converted into the compounds of the present invention. In addition, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an in vivo environment.

[0127] The terms used herein for compounds containing the aforementioned general structure have the following meanings:

[0128] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0129] The term "cyano" refers to -CN.

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

[0131] The term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group consisting of carbon atoms and hydrogen atoms, such as C 1-6 Alkyl groups, including but not limited to C 2-6 Alkyl, C 3-6Alkyl, etc., non-limiting examples of which include but are not limited to methyl, ethyl, propyl (including: 1-propyl or n-propyl, 2-propyl or isopropyl), butyl (including: 1-butyl or n-butyl, 2-methyl-1-propyl or isobutyl, 2-methyl-1-propyl or isobutyl, 1-methylpropyl or sec-butyl, 1,1-dimethylethyl or tert-butyl), pentyl (1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl), hexyl (1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl and 3,3-dimethyl-2-butyl); and C 2-6 Alkyl, that is, C 1-6 The alkyl group excludes the methyl group.

[0132] The term "alkenyl" refers to a linear or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one double bond. The alkenyl group may contain 2 to 20 carbon atoms, preferably 2 to 10 carbon atoms (i.e., C 2-10 Alkenyl), further preferably 2-8 carbon atoms (ie C 2-8 Alkenyl), more preferably 2-6 carbon atoms (ie C 2-6 alkenyl), 2-5 carbon atoms (ie C 2-5 alkenyl), 2-4 carbon atoms (ie C 2-4 alkenyl), 2-3 carbon atoms (ie C 2-3 alkenyl), 2 carbon atoms (i.e., C2 alkenyl), for example, "C 2-6 The term "alkenyl" refers to an alkenyl group, and the number of carbon atoms in the carbon chain is between 2 and 6 (i.e., 2, 3, 4, 5, or 6). Non-limiting examples of alkenyl groups include, but are not limited to, ethenyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 1,3-butadien-1-yl, 1,3-butadien-2-yl, and the like.

[0133] The term "alkynyl" refers to a straight or branched unsaturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms and having at least one triple bond. The alkynyl group may contain 2 to 20 carbon atoms, preferably 2 to 10 carbon atoms (i.e., C 2-10 Alkynyl), further preferably 2-8 carbon atoms (C 2-8 Alkynyl), more preferably 2-6 carbon atoms (ie C 2-6 Alkynyl), 2-5 carbon atoms (ie C 2-5 Alkynyl), 2-4 carbon atoms (ie C 2-4 Alkynyl), 2-3 carbon atoms (ie C 2-3alkynyl), 2 carbon atoms (i.e., C2 alkynyl), for example, "C 2-6 The term "alkynyl" refers to an alkynyl group, and the number of carbon atoms in the carbon chain is between 2 and 6 (i.e., 2, 3, 4, 5, or 6). Non-limiting examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, 1,3-butadiynyl, 1-pentynyl, 3-methyl-1-butynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 3-methyl-1-pentynyl, 4-methyl-1-pentynyl, 3,3-dimethyl-1-butynyl, 3-ethyl-1-butynyl, 1,3-hexadiynyl, 1,4-hexadiynyl, 3-methyl-1,4-pentadiynyl, 1,5-hexadiynyl, and the like.

[0134] The term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine and iodine). Examples of haloalkyl groups include halo-C 1-8 Alkyl, halogenated C 1-6 Alkyl or halogenated C 1-4 Alkyl groups include, but are not limited to, -CF3, -CH2Cl, -CH2CF3, -CCl2, CF3, etc.

[0135] The term "C 1-6 "Alkoxy" refers to an -O-alkyl group, wherein alkyl is as defined above. Examples of "alkoxy" as used herein include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, and tert-butoxy. "Alkoxy" also includes substituted alkoxy groups.

[0136] The term "C 1-6 "Alkylamino" refers to C 1-6 Monoalkyl-substituted amino, or two C 1-6 Alkyl-substituted amino, wherein alkyl is as defined above. As used herein, "C 1-6 Examples of "monoalkylamino" include, but are not limited to, methylamino, ethylamino, propylamino, cyclopropylamino, isopropylamino; "C 1-6 Examples of "dialkylamino" include, but are not limited to, dimethylamino, diethylamino, aziridine, azetidinyl, azopentyl, and azohexanyl.

[0137] The term "aromatic ring" refers to an all-carbon monocyclic ring or fused polycyclic ring of 6-14 carbon atoms with a completely conjugated π-electron system, including but not limited to a benzene ring, a naphthalene ring, an anthracene ring, etc., preferably a benzene ring.

[0138] The term "aryl" refers to an all-carbon monocyclic or condensed polycyclic group of 6-14 carbon atoms with a completely conjugated π-electron system, including but not limited to phenyl, naphthyl, anthracenyl, etc., preferably phenyl.

[0139] The term "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic ring (e.g., spirocyclic, bridged, etc.) containing 3-10 ring atoms, which is a non-aromatic structure; the polycyclic ring may be a non-aromatic ring in which all rings are non-aromatic, or at least one ring is aromatic and the remaining rings are non-aromatic. The aforementioned 3-10 ring atoms contain one or more (e.g., 2, 3, 4 or more) heteroatoms, and the rest are carbon atoms, and the heteroatoms are selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic ring may include the same or different heteroatoms in one or more rings, and the number of heteroatoms may be one or more. Non-limiting examples of "heterocycle" include, but are not limited to, aziridine, oxirane, thiirane, azetidine, oxetane, thietane, tetrahydrofuran, piperidine, piperazine, morpholine, thiomorpholine, etc.

[0140] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic (e.g., spirocyclic, bridged, etc.) group containing 3-10 ring atoms, which is a non-aromatic structure; the polycyclic ring may be all non-aromatic rings, or at least one ring may be aromatic and the remaining rings may be non-aromatic rings. The aforementioned 3-10 ring atoms contain one or more (e.g., 2, 3, 4 or more) heteroatoms, and the rest are carbon atoms, and the heteroatoms are selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic ring may include the same or different heteroatoms in one or more rings, and the number of heteroatoms may be one or more. Non-limiting examples of "heterocyclyl" include, but are not limited to, aziridine, oxirane, thiirane, azetidinyl, oxetanyl, thietanyl, furanyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, thiomorpholinyl, and the like.

[0141] The term "heteroaromatic ring" refers to an aromatic monocyclic or polycyclic ring (e.g., a fused ring) containing 5-14 ring atoms, wherein the aforementioned 5-14 ring atoms contain one or more (e.g., 2, 3, 4 or more) heteroatoms, and the rest are carbon atoms, wherein the heteroatoms are selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic ring may include the same or different heteroatoms in one or more rings, and the number of heteroatoms may be one or more. The aforementioned "heteroaromatic ring" preferably contains 5-14, 5-10, or 5-8 ring atoms, and more preferably contains 5-6 ring atoms. Non-limiting examples of "heteroaromatic ring" include, but are not limited to, tetrahydrofuran, thiophene, oxazole, thiazole, pyrrole, pyrazole, imidazole, pyridine, pyrimidine, pyrazinyl, pyridazine, quinoline, indole, benzofuran, benzothiophene, benzimidazole, benzopyridine, benzopyrimidine, benzopyrazine, and the like.

[0142] The term "heteroaryl" refers to an aromatic monocyclic or polycyclic (e.g., fused ring) group containing 5-14 ring atoms, wherein the aforementioned 5-14 ring atoms contain one or more (e.g., 2, 3, 4 or more) heteroatoms, and the remainder are carbon atoms, wherein the heteroatoms are selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic ring may include the same or different heteroatoms in one or more rings, and the number of heteroatoms may be one or more. The aforementioned "heteroaryl" preferably contains 5-14, 5-12, 5-10, or 5-8 ring atoms, and more preferably contains 5-6 ring atoms. Non-limiting examples of "heteroaryl" include, but are not limited to, furyl, thienyl, oxazolyl, thiazolyl, pyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolyl, indolyl, benzofuranyl, benzothienyl, benzimidazolyl, benzopyridinyl, benzopyrimidinyl, benzopyrazinyl, and the like.

[0143] The present invention also provides a pharmaceutical composition comprising at least one compound as described above or a pharmaceutically acceptable salt thereof as an active ingredient, and at least one or more pharmaceutically acceptable carriers.

[0144] The "pharmaceutical composition" of the present invention refers to a preparation of one or more compounds of the present invention or their salts and a carrier generally accepted in the art for delivering biologically active compounds to an organism (e.g., a human). The purpose of a pharmaceutical composition is to facilitate administration and delivery to an organism.

[0145] In particular, the pharmaceutical composition provided by the present invention adopts an inhalable drug delivery method.

[0146] The term "inhalable" as used herein refers to a compound that can be administered via the airways or lungs, absorbed by the lung epithelial cells or respiratory mucosa, and directly enter the bloodstream. "Inhalable" indicates that it can be administered by inhalation, but is not limited to inhalation. It is also suitable for routes of administration such as rectal administration, topical administration (including transdermal administration), vaginal administration, or parenteral administration (including subcutaneous injection, intramuscular injection, intravenous injection, or intradermal injection). These formulations can be prepared by any method known in the art of pharmacy, such as by mixing the active ingredient with a carrier or excipient.

[0147] The present invention also provides a method for preparing the aforementioned compound or pharmaceutical composition for use in preventing or treating diseases associated with PI3Kδ kinase activity.

[0148] Preferably, the aforementioned medicament is used for preventing or treating allergic diseases and inflammatory diseases; in particular, it plays a role in preventing or treating asthma, COPD and autoimmune diseases associated with PI3Kδ deficiency via inhalation administration.

[0149] More preferably, the aforementioned allergic diseases and inflammatory diseases are selected from asthma of any type or cause, including but not limited to intrinsic asthma, exogenous asthma, mild asthma, moderate asthma, severe asthma, bronchitis asthma, exercise-induced asthma, occupational asthma, Th2 asthma and non-Th2 asthma, wheezing infant syndrome, acute lung injury, chronic obstructive pulmonary disease, chronic bronchitis, emphysema, adult-onset / acute respiratory distress syndrome and other respiratory diseases; and autoimmune diseases such as rheumatoid arthritis, osteoarthritis, lupus erythematosus, psoriasis, atopic dermatitis, multiple sclerosis and the like.

[0150] In particular, other inflammatory or obstructive airway diseases to which the present invention is applicable are selected from pneumoconiosis of any type or origin, including but not limited to asbestosis, fume congestion, siderosis, hairline pneumoconiosis, and byssinosis.

[0151] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0152] Based on the target design of PI3Kδ inhibitors and the principle of bioisosterism, the present invention has developed a series of aromatic ring thiazine derivatives with novel structures that can be administered by inhalation. Relevant biological experiments have been carried out. The results show that some of the compounds developed by the present invention have high selectivity for PI3Kδ, significant inhibitory effects, and good safety. Moreover, after inhalation delivery, some of the compounds can be well retained in the lungs and have excellent inhalation pharmacokinetic properties. They can be used as inhaled drugs for the prevention or treatment of respiratory diseases such as asthma and COPD. BRIEF DESCRIPTION OF THE DRAWINGS

[0153] FIG1 is a graph showing the effect of compound BIOS-B12 on ovalbumin-induced airway hyperresponsiveness in mice.

[0154] FIG2 is a graph showing the effect of compound BIOS-B12 on ovalbumin-induced cell accumulation in the mouse airway. DETAILED DESCRIPTION

[0155] The following are specific embodiments of the present invention to further describe the technical solution of the present invention, but the scope of protection of the present invention is not limited to these embodiments. Any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the scope of protection of the present invention.

[0156] The Chinese names corresponding to the English abbreviations of the compounds used in the reaction process of the examples of the present invention are as follows:

[0157] Boc2O: di-tert-butyl carbonate;

[0158] DMAP: 4-dimethylaminopyridine;

[0159] Pd(PPh3)4: tetrakistriphenylphosphine palladium;

[0160] PPh3: triphenylphosphine;

[0161] DIPEA: diisopropylethylenediamine;

[0162] DEAD: diethyl azodicarboxylate;

[0163] DMF: N,N-dimethylformamide;

[0164] EA: ethyl acetate;

[0165] PdCl2(CH3CN)2: palladium chloride bis(acetonitrile);

[0166] Pd(amphos)2Cl2: dichlorodi-tert-butyl-(4-dimethylaminophenyl)phosphine palladium(II);

[0167] HATU: 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate;

[0168] DMSO: dimethyl sulfoxide;

[0169] X-phos: 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl.

[0170] In addition, all operations involving raw materials that are easily oxidized or hydrolyzed are carried out under nitrogen protection. Unless otherwise specified, the raw materials used in the present invention are commercially available raw materials and can be used directly without further purification.

[0171] The reaction starting materials and common intermediates involved in the embodiments of the present invention can be purchased commercially or prepared in-house. The preparation process of the starting materials and common intermediates that need to be prepared in-house is described in detail as follows:

[0172] 1. Synthesis of intermediate 2-bromo-6-chlorobenzenesulfonyl chloride (BIOS-A1-1):

[0173] Under ice-cooling, 2-bromo-6-chloroaniline (7g, 35mmol) was added to concentrated hydrochloric acid (30mL), followed by the slow addition of 7mL of sodium nitrite solution (2.8g, 41mmol). In a separate reaction flask, copper sulfate pentahydrate (0.93g, 3.7mmol) and 32mL of concentrated hydrochloric acid were added, followed by the slow dropwise addition of 20mL of sodium bisulfite solution (7.05g, 67.8mmol) and the above solution under ice-cooling. After the addition was complete, the mixture was allowed to warm to room temperature and react for 2h until the reaction was complete. The reaction solution was extracted with dichloromethane (80mL*3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to yield crude BIOS-A1-1 (7.0g, 24.13mmol), which was carried on to the next step without purification.

[0174] 2. Synthesis of the intermediate tert-butyl 4-amino-1H-pyrazole-1-carboxylate BIOS-A1-2:

[0175] 4-Nitro-1H-pyrazole (1.1 g, 10 mmol) was dissolved in dichloromethane, and Boc2O (2.2 g, 10 mmol) and DMAP (110 mg, 1 mmol) were added, respectively. The mixture was stirred at room temperature for 2 hours. After completion, the reaction was quenched by the addition of ammonium chloride solution. The organic phase was concentrated and purified by flash column chromatography to obtain tert-butyl 4-nitro-1H-pyrazole-1-carboxylate (1.7 g, 8 mmol). This intermediate was dissolved in methanol, purged with nitrogen, and palladium on carbon was added. A hydrogen balloon was added, and the atmosphere was replaced with hydrogen three times. The reaction was allowed to react overnight. After completion, the mixture was filtered through a pad of celite, and the filtrate was concentrated to obtain BIOS-A1-2 (0.9 g, 5 mmol) as a pale yellow solid.

[0176] 3. Synthesis of intermediate (S)-2-(but-3-yn-2-yl)isoindoline-1,3-dione A-004:

[0177] Phthalimide (5.0 g, 34.3 mmol), (R)-3-butyn-2-ol (2.0 g, 28.6 mmol), and PPh3 (9.0 g, 34.3 mmol) were dissolved in 50 mL of anhydrous tetrahydrofuran. DEAD (6.0 g, 34.3 mmol) was placed in a constant pressure dropping funnel and added dropwise at 0°C. After addition, the mixture was warmed to room temperature and stirred for 27 hours. After the starting material disappeared, the solvent was evaporated and concentrated to obtain a yellow oil. Purification and separation by flash column chromatography (PE:EA = 20:1) afforded 5.4 g of a white granular powder in an 80% yield.

[0178] Example 1. Preparation of target compound BIOS-A1

[0179] Step 1: Synthesis of intermediate tert-butyl 4-((2-bromo-6-chlorophenyl)sulfonylamino)-1H-pyrazole-1-carboxylate (BIOS-A1-3)

[0180] Intermediate BIOS-A1-1 (1.7 g, 6 mmol) was added to a dichloromethane solution containing BIOS-A1-2 (0.9 g, 5 mmol) and pyridine (2 mL). The mixture was stirred at room temperature overnight. After completion, the reaction solution was washed 3–5 times with ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated. Purification by flash column chromatography afforded BIOS-A1-3 (1.3 g, 3 mmol) as a light gray solid in a 60% yield.

[0181] ESI-MS: m / z = 436 [M+H]+ .

[0182] Step 2: Synthesis of intermediate (S)-3-(1-phthalamidoethyl)-8-chloro-2-(1H-pyrazol-4-yl)-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A1-4):

[0183] Intermediate BIOS-A1-3 (1.3 g, 3 mmol), A-004 (720 mg, 3.6 mmol), and Pd(PPh3)4 (330 mg, 10 mol%) were placed in a dry, sealed tube. The atmosphere was replaced with N2 three times. Acetonitrile (30 mL) and DIPEA (1.2 g, 9 mmol) were added under positive pressure, and the mixture was stirred overnight at 90°C. After the starting material disappeared, the solvent was evaporated to obtain a black oil. BIOS-A1-4 (454 mg, 1 mmol) was purified by flash column chromatography to obtain a pale yellow solid in a 33% yield.

[0184] ESI-MS: m / z = 455 [M+H] + .

[0185] Step 3: Synthesis of intermediate (S)-3-(1-aminoethyl)-8-chloro-2-(1H-pyrazol-4-yl)-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A1-5)

[0186] Intermediate BIOS-A1-4 (454 mg, 1 mmol) and hydrazine hydrate (85%, 0.2 mL) were added to 10 mL of ethanol solution and stirred at 80°C for 2 h. After the reaction was complete, the mixture was cooled thoroughly, filtered, and the filtrate was concentrated to obtain 200 mg of a white solid, which was directly carried to the next step without purification.

[0187] ESI-MS: m / z = 325 [M+H] + .

[0188] Step 4: Synthesis of (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-(1H-pyrazol-4-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile (BIOS-A1)

[0189] Intermediate BIOS-A1-5 (65 mg, 0.2 mmol), 2-amino-6-chloro-4-methylpyrimidine-5-carbonitrile (50 mg, 0.3 mmol), DIPEA (130 mg, 1.0 mmol), and DMSO (2.0 mL) were added sequentially to a single-necked flask and reacted at 90°C for 8 h. The mixture was concentrated under reduced pressure and extracted with an appropriate amount of dichloromethane. The product was washed sequentially with saturated aqueous NaHCO3, dilute hydrochloric acid (0.5 N), and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by preparative liquid chromatography to obtain the target product BIOS-A1 (46 mg) in a 50% yield.

[0190] ESI-MS: m / z = 457 [M+H] + .

[0191] 1 H NMR (400MHz, CDCl3) δ8.05 (s, 1H), 7.61 (s, 2H), 7.53-7.44 (m, 2H), 7.31 (dd, J = 6.9, 2.1Hz, 1H) ,6.53(s,1H),5.56(d,J=7.2Hz,1H),4.83(t,J=7.1Hz,1H),2.63(s,3H),1.54(d,J=7.0Hz,3H).

[0192] Example 2. Preparation of target compound BIOS-A2

[0193] Step 1: Synthesis of intermediate 2-bromo-6-chloro-N-(pyridin-3-yl)benzenesulfonamide (BIOS-A2-1):

[0194] Intermediate BIOS-A1-1 (1.4 g, 5 mmol) was dissolved in dichloromethane (20 mL), and pyridine (2 mL) and 3-aminopyridine (450 mg, 5 mmol) were added. The mixture was stirred at room temperature overnight until the reaction was complete. The reaction was quenched with water, and the organic phase was separated. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. BIOS-A2-1 (0.7 g) was isolated by flash column chromatography as a white solid in a 50% yield.

[0195] ESI-MS: m / z = 347 [M+H] + .

[0196] Step 2: Synthesis of intermediate (S)-2-(1-(8-chloro-1,1-dihydroxy-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)isoindoline-1,3-dione (BIOS-A2-2):

[0197] Intermediate BIOS-A2-1 (350 mg, 1 mmol), A-004 (240 mg, 1.2 mmol), and Pd(PPh3)4 (110 mg, 10 mol%) were placed in a dry, sealed tube. The atmosphere was replaced with N2 three times. Acetonitrile (20 mL) and DIPEA (400 mg, 3 mmol) were added under positive pressure, and the mixture was stirred overnight at 90°C. After the starting material disappeared, the solvent was evaporated to obtain a black oil. BIOS-A2-2 (200 mg) was purified by flash column chromatography to obtain a light yellow solid in a 43% yield.

[0198] ESI-MS: m / z = 466 [M+H] + .

[0199] Step 3: Synthesis of intermediate (S)-3-(1-aminoethyl)-8-chloro-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A2-3):

[0200] Intermediate BIOS-A2-2 (200 mg, 0.43 mmol) and hydrazine hydrate (85%, 0.3 ml) were added to an ethanol solution (10 mL). The mixture was stirred at 80°C for 2 h. After the reaction was complete, the mixture was cooled thoroughly, filtered, and the filtrate was concentrated to obtain a white solid BIOS-A2-3 (100 mg). This was carried on to the next step without purification.

[0201] ESI-MS: m / z = 336 [M+H] + .

[0202] Step 4: Synthesis of (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile (BIOS-A2)

[0203] Intermediate BIOS-A2-3 (100 mg, 0.3 mmol), 2-amino-6-chloro-4-methylpyrimidine-5-carbonitrile (51.0 mg, 0.3 mmol), DIPEA (156.0 μL, 0.9 mmol), and DMSO (2.0 mL) were added sequentially to a single-necked flask and reacted at 90°C for 8 h until the starting materials reacted completely. The mixture was concentrated under reduced pressure and extracted with an appropriate amount of dichloromethane. The product was washed sequentially with saturated aqueous NaHCO3, dilute hydrochloric acid (0.5 N), and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by preparative liquid chromatography to obtain the target product BIOS-A2 (40 mg) in a 28% yield.

[0204] ESI-MS: m / z = 468 [M+H] + .

[0205] 1 H NMR(400MHz, DMSO-d6)δ8.58(dd,J=4.4,1.2Hz,1H),8.38(dd,J=2.8,0.8Hz,1H),7.76-7.62(m,4H),7.56(d,J=7.6Hz, 1H),7.52-7.49(m,1H),7.19(s,1H),6.94(s,1H),6.29(s,1H),4.56-4.49(m,1H),2.26(s,3H),1.42(d,J=6.8Hz,3H).

[0206] Example 3. Preparation of target compound BIOS-A3

[0207] Step 1: Synthesis of intermediate 2-bromo-6-chloro-N-(pyridin-4-yl)benzenesulfonamide (BIOS-A3-1)

[0208] The intermediate 2-bromo-6-chlorobenzenesulfonyl chloride (1.4 g, 5 mmol) was dissolved in dichloromethane (20 mL), and pyridine (2 mL) and 3-aminopyridine (450 mg, 5 mmol) were added. The mixture was stirred at room temperature overnight until the reaction was complete. The reaction was quenched with water, and the layers were separated. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. BIOS-A3-1 (1.0 g) was isolated by flash column chromatography as a white solid in a 60% yield.

[0209] ESI-MS: m / z = 347 [M+H] + .

[0210] Step 2: Synthesis of intermediate (S)-2-(1-(8-chloro-1,1-dihydroxy-2-(pyridin-4-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)isoindoline-1,3-dione (BIOS-A3-2)

[0211] Intermediate BIOS-A3-1 (700 mg, 2 mmol), an alkynamide derivative (480 mg, 2.4 mmol), and Pd(PPh3)4 (220 mg, 10 mol%) were placed in a dry, sealed tube. The atmosphere was replaced with nitrogen three times. Acetonitrile (30 mL) and DIPEA (800 mg, 6 mmol) were added under positive pressure and stirred overnight at 90°C. After the starting material disappeared, the solvent was evaporated to obtain a black oil. Flash column chromatography afforded 200 mg of a light yellow solid (22% yield).

[0212] ESI-MS: m / z = 466 [M+H] + .

[0213] Step 3: Synthesis of intermediate (S)-3-(1-aminoethyl)-8-chloro-2-(pyridin-4-yl)-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A3-3)

[0214] Intermediate BIOS-A3-2 (200 mg, 0.43 mmol) and hydrazine hydrate (85%, 0.3 mL) were added to an ethanol solution (10 mL). The mixture was stirred at 80°C for 2 h. After the reaction was complete, the mixture was cooled thoroughly, filtered, and the filtrate was concentrated to obtain 100 mg of a white solid, which was directly carried to the next step without purification.

[0215] ESI-MS: m / z = 336 [M+H] + .

[0216] Step 4: Synthesis of (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-(pyridin-4-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile (BIOS-A3)

[0217] Intermediate BIOS-A3-3 (100 mg, 0.3 mmol), 2-amino-6-chloro-4-methylpyrimidine-5-carbonitrile (51.0 mg, 0.3 mmol), DIPEA (156.0 μL, 0.9 mmol), and DMSO (2.0 mL) were added sequentially to a single-necked flask and reacted at 90°C for 8 h. The mixture was concentrated under reduced pressure and extracted with an appropriate amount of dichloromethane. The product was washed sequentially with saturated aqueous NaHCO₃, dilute hydrochloric acid (0.5 N), and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by HPLC to obtain the target product BIOS-A3 (30 mg) in a 21% yield.

[0218] ESI-MS: m / z = 468 [M+H] + .

[0219] 1 H NMR (400MHz, DMSO-d6) δ9.85(d,J=7.4Hz,1H),8.58(dd,J=7.7,2.0Hz,1H),8.49(dd,J=7.6,2.0Hz,1H),8.40-8.23(m,2H),7.38(t,J=7.7Hz,1H),7.34 -7.16(m,3H),7.12(dd,J=7.9,2.7Hz,1H),6.09(s,1H),4.61-4.54(m,1H),2.54(s,3H),1.54(d,J=6.7Hz,3H).

[0220] Example 4. Preparation of target compound BIOS-A4

[0221] Step 1: Preparation of 2-bromo-6-fluorobenzenesulfonyl chloride (BIOS-A4-2)

[0222] Under ice, 2-bromo-6-fluoroaniline (3.85 g, 20.4 mmol) was added to 20 mL of concentrated hydrochloric acid, followed by the slow addition of 7 mL of sodium nitrite solution (2.8 g, 41 mmol). In a separate reaction flask, copper sulfate pentahydrate (0.93 g, 3.7 mmol) and 32 mL of concentrated hydrochloric acid were added, followed by the slow dropwise addition of 20 mL of sodium bisulfite solution (7.05 g, 67.8 mmol) and the above solution under ice. After the addition was complete, the mixture was allowed to warm to room temperature and react for 2 h until the reaction was complete. The reaction solution was extracted with dichloromethane (80 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to yield crude BIOS-A4-2 (5.0 g). This product was carried on to the next step without purification.

[0223] LC-MS: 272.0 [M+H] + .

[0224] Step 2: Preparation of 2-bromo-6-fluoro-N-(pyridin-3-yl)benzenesulfonamide (BIOS-A4-3)

[0225] Under ice, 3-aminopyridine (2.3 g, 24.1 mmol) was dissolved in 40 mL of pyridine, and BIOS-A4-2 (5.0 g) was added dropwise. The mixture was warmed to room temperature and allowed to react for 2 h until the starting material was completely reacted. The reaction mixture was concentrated under reduced pressure, then added with 50 mL of dichloromethane. The mixture was washed twice with 1N HCl (50 mL x 2) and water (50 mL x 2). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain BIOS-A4-3 (3.6 g) in a 59% yield.

[0226] LC-MS: 331.0 [M+H] + .

[0227] Step 3: Preparation of (S)-2-(1-(8-fluoro-1,1-dihydroxy-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)isoindoline-1,3-dione (BIOS-A4-4)

[0228] BIOS-A4-3 (3.6 g, 10.9 mmol), intermediate A-004 (3.3 g, 16.4 mmol), and Pd(PPh3)4 (5 mol%) were placed in a dry, sealed tube. The atmosphere was replaced with N2 three times. 50 mL of acetonitrile and DIPEA (32.7 mmol) were added under positive pressure, and the mixture was stirred overnight at 90°C. After the starting material disappeared, the solvent was evaporated to obtain a black oil. BIOS-A4-4 was isolated by flash column chromatography as a pale yellow solid in a 71% yield.

[0229] LC-MS: 450.0 [M+H] + .

[0230] Step 4: Preparation of (S)-3-(1-aminoethyl)-8-fluoro-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A4-5)

[0231] BIOS-A4-4 and hydrazine hydrate (85%, 1 mL, 15.5 mmol) were added to a 50 mL ethanol solution, thoroughly cooled, and the mixture was replaced with nitrogen three times. The reaction was stirred at 80°C for 2 h. After the reaction was complete, the mixture was thoroughly cooled to precipitate a large amount of white flocculent precipitate, which was filtered and concentrated to obtain a white solid. Ethanol solution was added to the mixture, cooled, filtered, and the filtrate was further concentrated. Ethyl acetate was added, cooled, filtered, and the filtrate was dried to dryness to obtain the crude yellow oil BIOS-A4-5. This product was directly carried to the next step without purification, with a yield of 85%.

[0232] LC-MS: 320.0 [M+H] + .

[0233] Step 5: Preparation of (S)-2-amino-4-((1-(8-fluoro-1,1-dihydroxy-2-(pyridin-3-yl)-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile (BIOS-A4)

[0234] Intermediate BIOS-A4-5 (320 mg, 1 mmol) was dissolved in 3 mL of DMSO, and 2-amino-4-chloro-6-methylpyrimidine-5-carbonitrile (200 mg, 1.2 mmol) and DIPEA (387 mg, 3 mmol) were added. The atmosphere was replaced with nitrogen three times, and the mixture was stirred at 90°C overnight. 10 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The target product, BIOS-A4 (280 mg), was isolated and purified by column chromatography as a yellow solid in a 62% yield.

[0235] LC-MS:452.0[M+H] + .

[0236] 1 H NMR (400MHz, DMSO-d6) δ8.58(d,J=4.8Hz,1H),8.37(d,J=2.6Hz,1H),7.82-7.72(m,2H),7.64- 7.40(m,4H),6.98(s,2H),6.28(s,1H),4.57-4.49(m,1H),2.26(s,3H),1.42(d,J=7.0Hz,3H).

[0237] Example 5. Preparation of target compound BIOS-A5

[0238] Referring to Example 4, the compound 3-amino-pyridine in step 2 of Example 4 was replaced with 5-fluoropyridin-3-amine, and other operations remained unchanged to obtain compound BIOS-A5.

[0239] LC-MS:470.1[M+H] + .

[0240] Example 6. Preparation of target compound BIOS-A6

[0241] Referring to Example 4, the compound BIOS-A4-1 in step 1 of Example 4 was replaced with 2-bromo-6-chloroaniline, and the 3-amino-pyridine in step 2 was replaced with 5-fluoropyridin-3-amine. Other operations remained unchanged to obtain compound BIOS-A6 in a yield of 63%.

[0242] LC-MS:486.0[M+H] + .

[0243] 1 H NMR (400MHz, DMSO-d6) δ8.62(d,J=2.6Hz,1H),8.23(d,J=1.8Hz,1H),7.81-7.60(m,4H),7.49(d ,J=7.7Hz,1H),7.00(s,2H),6.29(s,1H),4.69-4.62(m,1H),2.25(s,3H),1.44(d,J=7.0Hz,3H).

[0244] Example 7. Preparation of target compound BIOS-A7

[0245] Step 1: Synthesis of intermediate N-benzyl-2-bromo-6-chlorobenzenesulfonamide (BIOS-A7-1)

[0246] The intermediate 2-bromo-6-chlorobenzenesulfonyl chloride (1.45 g, 5 mmol) was added to a dichloromethane solution containing benzylamine (520 mg, 5 mmol) and triethylamine (1 mL). The mixture was stirred at room temperature overnight. After completion of the reaction, the reaction solution was washed 3-5 times with ammonium chloride solution. The organic phase was dried over anhydrous sodium sulfate and concentrated. Purification by flash column chromatography afforded BIOS-A7-1 (1.4 g) as a light gray solid in 80% yield.

[0247] ESI-MS: m / z = 359 [M+H] + .

[0248] Step 2: Synthesis of intermediate S-2-(1-(2-benzyl-8-chloro-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-3-yl)ethyl)isoindoline-1,3-dione (BIOS-A7-2)

[0249] BIOS-A7-1 (700 mg, 2 mmol), an alkynylamine derivative (480 mg, 2.4 mmol), and Pd(PPh3)4 (220 mg, 10 mol%) were placed in a dry, sealed tube. The atmosphere was replaced with N2 three times. Acetonitrile (30 mL) and DIPEA (800 mg, 6 mmol) were added under positive pressure, and the mixture was stirred overnight at 90°C. After the starting material disappeared, the solvent was evaporated to obtain a black oil. Flash column chromatography afforded BIOS-A7-2 (280 mg) as a pale yellow solid in a 30% yield.

[0250] ESI-MS: m / z = 479 [M+H] + .

[0251] Step 3: Synthesis of (S)-3-(1-aminoethyl)-2-benzyl-8-chloro-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A7-3)

[0252] Intermediate BIOS-A7-2 (280 mg, 0.6 mmol) and hydrazine hydrate (85%, 1.0 ml) were added to 15 mL of ethanol. The reaction was stirred at 80°C for 2 h. After TLC indicated completion of the reaction, the product was cooled, filtered, and the filtrate was concentrated to yield 100 mg of a white solid, which was carried on to the next step without purification.

[0253] ESI-MS: m / z = 349 [M+H] + .

[0254] Step 4: Synthesis of (S)-2-amino-4-((1-(2-benzyl-8-chloro-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-3-yl)ethyl)amino)-6-methylpyrimidine-5-carbonitrile (BIOS-A7)

[0255] Intermediate BIOS-A7-3 (100 mg, 0.3 mmol), 2-amino-6-chloro-4-methylpyrimidine-5-carbonitrile (66 mg, 0.4 mmol), DIPEA (130 mg, 1.0 mmol), and DMSO (2.0 mL) were added sequentially to a single-necked flask and reacted at 90°C for 8 h until the starting materials reacted completely. The mixture was concentrated under reduced pressure and extracted with an appropriate amount of dichloromethane. The product was washed sequentially with saturated aqueous NaHCO3, dilute hydrochloric acid (0.5 N), and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by HPLC to obtain the desired product, BIOS-A7 (48 mg), in a 33% yield.

[0256] ESI-MS: m / z = 481 [M+H] + .

[0257] 1 H NMR (400MHz, DMSO-d6) δ8.00-7.72(m,3H),7.36(m,4H),7.27(m,2H),7.08(d,J=8.0Hz,1H),6.70(s,1H),5.74(d,J =10.4Hz,1H),5.46(d,=18.2Hz,1H),5.18(d,J=18.2Hz,1H),5.11-5.05(m,1H),2.22(s,3H),0.70(d,J=6.5Hz,3H).

[0258] Example 8. Preparation of target compound BIOS-A8

[0259] BIOS-A6 (100 mg, 0.2 mmol) was dissolved in acetonitrile (2 mL), and PdCl2(CH3CN)2 (13 mg, 0.2 eq), X-phos (72 mg, 0.6 eq), cesium carbonate (212 mg, 3 eq), and N,N-bis(2-methoxyethyl)hex-5-ynamide (136 mg, 3 eq) were added. The atmosphere was replaced with nitrogen three times, and the mixture was stirred at reflux overnight. 10 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. BIOS-A8 (70 mg) was purified by silica gel column chromatography to obtain a yellow solid in a 50% yield.

[0260] LC-MS: 691.2[M+H] + .

[0261] Example 9. Preparation of target compound BIOS-B1

[0262] Step 1: Preparation of 2-chloro-6-methylbenzenesulfonyl chloride (BIOS-B1-2)

[0263] Under ice-cooling, 2-chloro-6-methylaniline (2.9 g, 20.4 mmol) was added to 20 mL of concentrated hydrochloric acid, followed by the slow addition of 7 mL of sodium nitrite solution (2.8 g, 41 mmol). In a separate reaction flask, copper sulfate pentahydrate (0.93 g, 3.7 mmol) and 32 mL of concentrated hydrochloric acid were added, followed by the slow dropwise addition of 20 mL of sodium bisulfite solution (7.05 g, 67.8 mmol) and the above solution under ice-cooling. After the addition was complete, the mixture was allowed to warm to room temperature and react for 2 h. The reaction solution was extracted with dichloromethane (80 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to yield crude BIOS-B1-2 (4.0 g). This product was carried on to the next step without purification.

[0264] LC-MS: 225.0 [M+H] + .

[0265] Step 2: Preparation of 2-chloro-6-methyl-N-phenylbenzenesulfonamide (BIOS-B1-3)

[0266] Under ice, aniline (2.5 g, 24.1 mmol) was dissolved in 40 mL of pyridine, and BIOS-B1-2 (4.0 g) was added dropwise. After the addition was complete, the mixture was warmed to room temperature and allowed to react for 2 hours until the starting material was completely reacted. The reaction mixture was concentrated under reduced pressure, then added with 50 mL of dichloromethane. The mixture was washed twice with 1N HCl (50 mL x 2) and water (50 mL x 2). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography to obtain BIOS-B1-3 (3.6 g) in a 72% yield.

[0267] LC-MS: 225.0 [M+H] + .

[0268] Step 3: Preparation of (S)-2-(1-(8-chloro-2-(5-fluoropyridin-3-yl)-1,1-dihydroxy-2H-benzo[e][1,2]thiazin-3-yl)ethyl)isoindoline-1,3-dione (BIOS-B1-4)

[0269] A solution of intermediate BIOS-B1-3 (1 g, 3.5 mmol) in anhydrous tetrahydrofuran (10 mL) was cooled to -78°C, followed by the addition of a 2.5 M solution of n-butyllithium in hexane (4.3 mL). The reaction mixture was stirred for 30 minutes. In a separate flask, a solution of tert-butyl (S)-(1-(methoxy(methyl)amino)-1-carbonylbutan-2-yl)carbamate (1.1 g, 4.55 mmol) in anhydrous tetrahydrofuran (10 mL) was cooled. A solution of isopropylmagnesium chloride in tetrahydrofuran (2.5 mL, 5.3 mmol) was slowly added at -78°C. The reaction mixture was stirred at the same temperature for 30 minutes, then added to the above reaction mixture. The mixture was stirred at -78°C for 1 hour. The reaction mixture was quenched with water (50 mL) and extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was carried on to the next step without purification.

[0270] LC-MS:467.0[M+H] + .

[0271] Step 4: Preparation of (S)-3-(1-aminopropyl)-8-chloro-2-phenyl-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-B1-5)

[0272] The crude intermediate BIOS-B1-4 and 10 mL of a 4 M hydrochloric acid solution in dioxane were added to a reaction flask and stirred at 100°C for 4 h. After TLC indicated completion of the reaction, the dioxane was removed under reduced pressure, a small amount of water was added, and the pH was adjusted to alkaline with potassium carbonate. The product was extracted with ethyl acetate (50 mL x 2). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The product BIOS-B1-5 (700 mg) was isolated by silica gel column chromatography.

[0273] LC-MS: 349.0 [M+H] + .

[0274] Step 5: Preparation of (S)-2-amino-4-((1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)propyl)amino)-6-methylpyrimidine-5-carbonitrile (BIOS-B1)

[0275] Intermediate BIOS-B1-5 (700 mg, 2 mmol) was dissolved in 7 mL of DMSO, followed by the addition of 2-amino-4-chloro-6-methylpyrimidine-5-carbonitrile (400 mg, 2.4 mmol) and DIPEA (774 mg, 6 mmol). The atmosphere was replaced with nitrogen three times and stirred at 90°C overnight until the reaction was complete. The reaction mixture was added with 10 mL of water and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The target product, BIOS-B1 (720 mg), was isolated and purified by silica gel column chromatography to obtain a yellow solid in a 69% yield.

[0276] LC-MS:481.0[M+H] + .

[0277] 1 H NMR (400MHz, CDCl3) δ7.52-7.43(m,2H),7.41-7.30(m,4H),7.26(s,1H),7.24-7.18(m,2H),6.61(s,1H), 5.29(d,2H),4.65-4.59(m,1H),2.42(s,3H),2.08-1.98(m,1H),1.81-1.71(m,1H),0.99(t,J=7.4Hz,3H).

[0278] Example 10. Preparation of target compound BIOS-B2

[0279] Referring to Example 9, the compound B-004-1 in step 3 of Example 9 was replaced with tert-butyl (S)-(4,4-difluoro-1-(methoxy(methyl)amino)-1-carbonylbutan-2-yl)carbamate, and the other operations remained unchanged to obtain compound BIOS-B2.

[0280] LC-MS: 517.1[M+H] + .

[0281] Example 11. Preparation of target compound BIOS-B3

[0282] Referring to Example 9, the compound B-004-1 in step 3 of Example was replaced with tert-butyl (S)-(1-cyclobutyl-2-(methoxy(methyl)amino)-2-carbonylethyl)carbamate, and the other operations remained unchanged to obtain the target compound BIOS-B3 (25 mg) as a white powder.

[0283] LC-MS: 507.2[M+H] + .

[0284] Example 12. Preparation of target compound BIOS-B4

[0285] Referring to Example 9, the compound B-004-1 in step 3 of Example was replaced with tert-butyl (S)-(1-(methoxy(methyl)amino)-3-methyl-1-carbonylbutan-2-yl)carbamate, and the other operations remained unchanged to obtain the target compound BIOS-B4 as a white powder in a yield of 59%.

[0286] LC-MS:495.0[M+H] + .

[0287] 1 H NMR (400MHz, CDCl3) δ7.54-7.45(m,2H),7.45-7.33(m,4H),7.27-2.20(m,2H),6.59(s,1H),5.31(d,J =8.5Hz,1H),4.97(s,2H),4.61(dd,J=8.5,4.6Hz,1H),2.43(s,3H),1.28(s,1H),1.02(d,J=10.7,6H).

[0288] Example 13. Preparation of target compound BIOS-B5

[0289] Referring to Example 9, compound B-004-1 in step 3 of Example was replaced with tert-butyl (S)-(1-cyclopropyl-2-(methoxy(methyl)amino)-2-carbonylethyl)carbamate. Other operations remained unchanged to obtain the target compound BIOS-B5 as a white powder in a yield of 63%.

[0290] LC-MS:493.0[M+H] + .

[0291] 1 H NMR (400MHz, CDCl3) δ7.55-7.45(m,2H),7.40-7.33(m,4H),7.26 -7.19(m,2H),6.79(s,1H),5.54(d,J=7.2Hz,1H),5.05(s,2H),4.12(t,J=8.1 Hz,1H),2.41(s,3H),1.26-1.16(m,1H),0.81-0.62(m,2H),0.45-0.29(m,2H).

[0292] Example 14. Preparation of target compound BIOS-B6

[0293] Referring to Example 9, compound B-004-1 in step 3 of Example was replaced with tert-butyl (S)-6-(methoxy(methyl)carbamoyl)-5-azaspiro[2.4]heptane-5-carboxylate. Other operations remained unchanged to obtain compound BIOS-B6 in a yield of 25%.

[0294] ESI-MS: m / z = 519 [M+H] + .

[0295] 1 H NMR (400MHz, DMSO-d6) δ7.70-7.65(m,2H),7.64-7.57(m,1H),7.47-7.35(m,3H),7.32-7.22(m,2H),6.84(s,1H),4.86(d,J=6.0Hz,1H), 3.83(d,J=9.9Hz,1H), 3.67(d,J=9.8Hz,1H), 2.27(s,3H), 2.08(dd,J=12.4,8.3Hz,1H), 1.81(dd,J=12.4,2.4Hz,1H), 0.71-0.43(m,4H).

[0296] Example 15. Preparation of target compound BIOS-B7

[0297] Referring to Example 9, compound B-004-1 in step 3 of Example was replaced with tert-butyl (S)-2-(methoxy(methyl)carbamoyl)pyrrolidine-1-carboxylate. Other operations remained unchanged to obtain the target compound BIOS-B7 as a white powder in 82% yield.

[0298] LC-MS:493.1[M+H] + .

[0299] 1 H NMR(400MHz, CDCl3)δ7.52-7.38(m,5H),7.34 -7.29(m,3H),6.36(s,1H),5.14(s,2H),4.90-4.81(m,1H),4.22(d,J=8.1Hz,1H),3.9 2-3.82(m,1H),2.47(s,3H),2.24-2.13(m,2H),2.06-1.98(m,1H),1.94-1.83(m,1H).

[0300] Example 16. Preparation of target compound BIOS-B8

[0301] Referring to Example 9, the compound 2-amino-4-chloro-6-methylpyrimidine-5-carbonitrile in Step 5 of Example 9 was replaced with 2,4-diamino-6-chloropyrimidine-5-carbonitrile. Other operations remained unchanged to obtain the target compound BIOS-B8 as a white powder in a yield of 30%.

[0302] LC-MS:482.0[M+H] + .

[0303] 1 H NMR(400MHz,DMSO-d6)δ7.82-7.63(m,3H),7.40-7.34(m,3H),7.15-7.04(m,3H),6.70-6.65( m,2H),4.56(s,1H),4.22-4.17(m,2H),4.06(d,1H),1.84-1.77(m,2H),0.87(t,J=7.2Hz,3H).

[0304] Example 17. Preparation of target compound BIOS-B9

[0305] Referring to Example 9, the compound 2-chloro-6-methylaniline in Step 1 of Example 9 was replaced with 2-fluoro-6-methylaniline, and the compound aniline in Step 2 was replaced with pyridin-3-ylmethylamine. Other operations remained unchanged to obtain the target compound BIOS-B9 as a white powder.

[0306] ESI-MS: m / z = 480.1 [M+H] + .

[0307] Example 18. Preparation of target compound BIOS-B10

[0308] Referring to Example 9, the starting material 2-chloro-6-methylaniline in Step 1 of Example 9 was replaced with 2-fluoro-6-methylaniline (BIOS-B9-1) to produce compound BIOS-B9-1. At the same time, the compound aniline in Step 2 of Example 9 was replaced with (5-fluoropyridin-3-yl)methanamine, and the other operations remained unchanged to obtain compound BIOS-B10.

[0309] ESI-MS: m / z = 498.1 [M+H] + .

[0310] Example 19. Preparation of target compound BIOS-B11

[0311] BIOS-B1 (120 mg, 0.25 mmol) was dissolved in acetonitrile (2 mL), and PdCl2(CH3CN)2 (13 mg, 0.2 eq), X-phos (72 mg, 0.6 eq), cesium carbonate (212 mg, 2.6 eq), and BIOS-B11-1 (119 mg, 3 eq) were added. The atmosphere was replaced with nitrogen three times, and the mixture was stirred at reflux overnight. 10 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution (20 mL x 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. BIOS-B11 (70 mg) was purified by silica gel column chromatography to obtain a yellow solid in a yield of 46%.

[0312] LC-MS: 603.2[M+H] + .

[0313] 1 H NMR(600MHz,DMSO-d6)δ7.75-7.64(m,2H),7.62(dd,J=7.3,1.7Hz,1H),7.46-7.34(m,3H), 7.20-7.12(m,3H),6.84(s,1H),6.29-5.94(m,2H),4.53-4.48(m,1H),4.37(s,2H),3.60(d d,J=5.9,3.7Hz,2H),3.52(dd,J=5.8,3.7Hz,2H),3.48(dd,J=5.9,3.7Hz,2H),3.40(dd,J= 5.8, 3.7Hz, 2H), 3.22 (s, 3H), 2.25 (d, J = 1.9Hz, 3H), 1.91-1.79 (m, 2H), 0.80-0.74 (m, 3H).

[0314] Example 20. Preparation of target compound BIOS-B12

[0315] Referring to Example 19, the compound BIOS-B11-1 in the step of Example 19 was replaced with N,N-bis(2-methoxyethyl)hex-5-ynamide, and the other operations remained unchanged to obtain a light yellow powder compound BIOS-B12 with a yield of 67%.

[0316] LC-MS: 672.2[M+H] + .

[0317] 1H NMR (400MHz, CDCl3) δ7.52-7.43(m,2H),7.36(dd,J=5.2,1.9Hz,3H),7.31(dd,J=7.5,1.5Hz,1H),7. 22(dd,J=6.7,3.0Hz,2H),6.55(s,1H),5.21(d,J=7.9Hz,1H),4.95(s,2H),4.61-4.57(m,1H),3.55( t,J=5.6Hz,2H),3.52-3.48(m,4H),3.42-3.38(m,2H),3.32(s,3H),3.25(s,3H),2.57(t,J=7.4Hz,2 H), 2.51 (t, J = 6.6Hz, 2H), 2.39 (s, 3H), 1.95-1.90 (m, 2H), 1.76-1.71 (m, 2H), 0.97 (t, J = 7.4Hz, 3H).

[0318] Example 21. Preparation of target compound BIOS-B13

[0319] With reference to Example 3, BIOS-B' (ie, reference compound 2) was prepared by replacing 3-aminopyridine in step 1 of Example 3 with aniline.

[0320] Referring to Example 19, the compound BIOS-B1 in the steps of Example 19 was replaced by BIOS-B', and other operations remained unchanged to obtain the target compound BIOS-B13 as a light yellow powder with a yield of 51%.

[0321] LC-MS:589.1[M+H] + .

[0322] 1 H NMR (400MHz, CDCl3) δ7.56-7.54(m,1H),7.52-7.49(m,1H),7.38 -7.35(m,4H),7.24-7.21(m,2H),6.59(s,1H),5.28(d,J=7.4Hz,1H),4.98(s,2H),4.76-4.71(m,1H),4.43(s,2H),3.77(dd,J=5.7,3.7Hz,2H) ,3.67(dd,J=5.8,3.6Hz,2H),3.63(dd,J=5.7,3.8Hz,2H),3.54(dd,J=5.8,3.6Hz,2H),3.39-3.34(m,3H),2.40(s,3H),1.49(d,J=6.9Hz,3H).

[0323] Example 22. Preparation of target compound BIOS-B14

[0324] Referring to Example 19, the compound BIOS-B11-1 in the step of Example 19 was replaced with 4-ethynyl-1-methyl-1H-pyrazole, and the other operations remained unchanged to obtain the target compound BIOS-B14 as a light yellow powder with a yield of 73%.

[0325] LC-MS: 551.1[M+H] + .

[0326] 1 H NMR (400MHz, CDCl3) δ7.67-7.49(m,4H),7.42-7.32(m,4H),7.26 -7.23(m,2H),6.59(s,1H),5.23(d,J=7.8Hz,1H),4.97(s,2H),4.66-4.60(m,1H),3. 89(s,3H),2.42(s,3H),2.10-2.02(m,1H),1.81-1.73(m,1H),1.00(t,J=7.4Hz,3H).

[0327] Example 23. Preparation of target compound BIOS-B15

[0328] Referring to Example 19, compound BIOS-B11-1 in the step of Example 19 was replaced with 1-morpholinohex-5-yn-1-one, and other operations remained unchanged to obtain compound BIOS-B15 in a yield of 69%.

[0329] LC-MS: 626.2[M+H] + .

[0330] 1H NMR (600MHz, CDCl3) δ7.52-7.46(m,2H),7.41-7.36(m,3H),7.32(dd,J=6.7,2.4Hz,1H),7.23-7.18(m,2H) ,6.55(s,1H),5.22(d,J=7.9Hz,1H),4.97(s,2H),4.60-4.56(m,1H),3.63(t,J=4.9Hz,2H),3.58(dd,J=5.7 ,3.7Hz,2H),3.53(t,J=4.8Hz,2H),3.43(dd,J=5.8,3.8Hz,2H),2.60(dt,J=7.4,2.7Hz,2H),2.54(dd,J=7. 1,5.5Hz,2H),2.39(s,3H),2.04-1.97(m,1H),1.94-1.88(m,2H),1.76-1.71(m,1H),0.97(t,J=7.4Hz,3H).

[0331] Example 24. Preparation of target compound BIOS-B16

[0332] Referring to Example 19, the compound BIOS-B11-1 in the step of Example 19 was replaced with 1-(pyrrolidin-1-yl)hex-5-yn-1-one, and the other operations remained unchanged to obtain compound BIOS-B16 in a yield of 67%.

[0333] LC-MS: 610.2[M+H] + .

[0334] 1 H NMR (600MHz, CDCl3) δ7.52-7.45(m,2H),7.39-7.33(m,3H),7.32 -7.30(m,1H),7.23-7.19(m,2H),6.56(s,1H),5.21(d,J=7.9Hz,1H),4.97(s, 2H),4.62-4.57(m,1H),3.42(t,J=6.8Hz,2H),3.37(t,J=6.8Hz,2H),2.53(t,J =6.5Hz,2H),2.50(t,J=7.4Hz,2H),2.39(s,3H),2.04-1.98(m,1H),1.95-1.90 (m,2H),1.88-1.85(m,2H),1.83-1.79(m,2H),1.76-1.71(m,1H),0.98(t,3H).

[0335] Example 25. Preparation of target compound BIOS-B17

[0336] To a reaction flask were added BIOS-B1 (150 mg, 0.31 mmol), BIOS-B17-1 (53 mg, 0.34 mmol), potassium phosphate (131 mg, 0.62 mmol), Pd(amphos)2Cl2 (22 mg, 0.03 mmol), n-butanol:water = 2:1 (3 mL total), and 1 mL of acetonitrile. The atmosphere was replaced with nitrogen and the reaction was allowed to proceed at 80°C overnight. After completion of the reaction, the solvent was removed under reduced pressure, water was added, and the mixture was extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to afford the target compound BIOS-B17 as a bright-colored powder in an 83% yield.

[0337] LC-MS: 554.1[M+H] + .

[0338] H NMR (400MHz, CDCl3) δ8.16 (dd, J=5.2, 0.8Hz, 1H), 7.63 (t, J=7.7Hz, 1H), 7.50 (dd, J=7.9,1.2Hz,1H),7.38-7.29(m,4H),7.22-7.12(m,2H),7.02(d,J=5.9Hz,1H),6. 79(s,1H),6.69(s,1H),5.27(d,J=7.9Hz,1H),4.98(s,2H),4.66-4.59(m,1H),3.9 4(s,3H),2.42(s,3H),2.10-1.98(m,1H),1.85-1.71(m,1H),1.01(t,J=7.4Hz,3H).

[0339] Reference Compound 1. Synthesis of (S)-3-amino-N-(1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)ethyl)pyrazine-2-carboxamide

[0340] Step 1: Synthesis of intermediate 2-bromo-6-chloro-N-phenylbenzenesulfonamide (1-1)

[0341] The intermediate 2-bromo-6-chlorobenzenesulfonyl chloride (1.4 g, 5 mmol) was dissolved in dichloromethane (20 mL), and pyridine (2 mL) and aniline (450 mg, 5 mmol) were added. The mixture was stirred at room temperature overnight. The reaction was quenched with water, and the layers were separated. The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain compound 1-1 (1.4 g) as a white solid in 80% yield.

[0342] ESI-MS: m / z = 346 [M+H] + .

[0343] Step 2: Synthesis of intermediate (S)-2-(1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)ethyl)isoindoline-1,3-dione (1-2)

[0344] Intermediate 1-1 (700 mg, 2 mmol), an alkynamide derivative (480 mg, 2.4 mmol), and Pd(PPh3)4 (220 mg, 10 mol%) were placed in a dry, sealed tube. The atmosphere was replaced with N2 three times. Acetonitrile (30 mL) and DIPEA (800 mg, 6 mmol) were added under positive pressure and stirred overnight at 90°C. After the starting material disappeared, the solvent was evaporated to give a black oil. Flash column chromatography afforded 1-2 (280 mg) as a pale yellow solid in a 30% yield.

[0345] ESI-MS: m / z = 465 [M+H] + .

[0346] Step 3: Synthesis of intermediate (S)-3-(1-aminoethyl)-8-chloro-2-phenyl-2H-benzo[e][1,2]thiazine 1,1-dioxide (1-3)

[0347] Intermediate 1-2 (280 mg, 0.6 mmol) and hydrazine hydrate (85%, 0.8 mL) were added to an ethanol solution (20 mL). The reaction was stirred at 80°C for 2 h. After TLC indicated completion of the reaction, the mixture was cooled, filtered, and the filtrate was concentrated to yield 160 mg of a white solid, which was carried on to the next step without purification.

[0348] ESI-MS: m / z = 335 [M+H] + .

[0349] Step 4: Synthesis of (S)-3-amino-N-(1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)ethyl)pyrazine-2-carboxamide (reference compound 1)

[0350] Intermediate 1-3 (140 mg, 0.3 mmol), 3-aminopyrazine-2-carboxylic acid (56 mg, 0.4 mmol), HATU (190 mg, 0.5 mmol), and DIPEA (130 mg, 1 mmol) were added sequentially to DMF. Stirring was continued at room temperature for 2 hours. LC-MS indicated the reaction was complete. The reaction solution was concentrated and directly purified by preparative chromatography to afford 68 mg of the desired product as a white solid in a 50% yield.

[0351] ESI-MS: m / z = 456 [M+H]+ .

[0352] 1 H NMR (400MHz, DMSO-d6) δ8.89(d,J=8.4Hz,1H),8.23(d,J=2.3Hz,1H),7.86(d,J=2.3Hz,1H),7.74 -7.57(m,3H),7.55-7.35(m,5H),7.19-7.17(m,2H),6.94(s,1H),4.54-4.41(m,1H),1.43(d,J=7.0Hz,3H).

[0353] Example 26. ADP-Glo ​​of the Compounds of the Invention TM Kinase activity assay

[0354] ADP-Glo TM Kinase assay compounds inhibit PI3K kinase. ADP-Glo TM The assay consists of two steps. First, after the kinase reaction, an equal volume of ADP-Glo TM The first step is to add a kinase assay reagent, which terminates the kinase reaction and depletes any remaining ATP. In the second step, a kinase assay reagent is added, which simultaneously converts ADP to ATP and allows for measurement of newly synthesized ATP using a luciferase / luciferin reaction. The light generated is measured using a luminometer. Using an ATP to ADP conversion curve, fluorescence can be correlated to ADP concentration.

[0355] Experimental procedure: Dilute the test compound to an appropriate concentration series (approximately 6 to 8 concentrations) and prepare a source plate. Perform the kinase reaction according to the kit and read the fluorescence signal. Calculate the percentage inhibition of each reaction relative to the DMSO-treated control using the formula: Inhibition rate = (maximum fluorescence - sample fluorescence) / (maximum fluorescence - minimum fluorescence) / 100. Calculate the 50% inhibitory concentration (IC) from the concentration-response curve. 50 value).

[0356] Table 1: IC of PI3K kinase inhibition 50 (nM)

Note

[0357] As shown in Table 1, the test compounds showed significant inhibitory effects on PI3Kδ, but showed no or only low inhibitory effects on PI3Kα, PI3Kβ and PI3Kγ.

[0358] Example 27. Inhibitory effects of some compounds on p-AKT in Joke-1 cells

[0359] TR-FRET combines time-resolved fluorescence detection with fluorescence resonance energy transfer (FRET). In a FRET experiment, biomolecules (such as proteins) are labeled with fluorescent groups called donor and acceptor fluorophores. When the biomolecules interact, the distance between the donor and acceptor fluorophores is brought closer. At this point, if the donor is excited, it transfers its emitted light energy to the acceptor. The light emitted by the acceptor and donor has different wavelengths, which can be distinguished by a microplate reader, allowing the interaction between the biomolecules to be quantified.

[0360] 8 μL of cell suspension was added to the assay plate. 2 μL of anti-IgM (250 ng / mL) was added and incubated at 30°C for 60 minutes to stimulate JeKo-1 cells. Different concentrations of the compound (0.000152 μM, 0.000457 μM, 0.00137 μM, 0.00411 μM, 0.0123 μM, 0.0370 μM, 0.111 μM, 0.333 μM, 1 μM) were added to the cells and incubated at 37°C for 10 minutes. The p-AKT (downstream product of PI3K) in the cells was quantitatively measured using the Phospho-AKT (Ser473) kit (Cisbio) to determine the PI3Kδ inhibitory activity of the compound, IC 50 The results are shown in Table 2.

[0361] Table 2: IC of PI3K kinase inhibition 50 (nM)

Note

[0362] Example 28. Ovalbumin (OVA)-induced allergic asthma model in mice

[0363] Female Balb / c mice aged 6-8 weeks were intraperitoneally injected with ovalbumin sensitization solution (OVA 25μg / mouse) on days 0 and 14. In order to induce a local inflammatory response in the lungs, mice were stimulated with a 1% OVA solution by aerosolization on days 21-25, once a day for 30 minutes each time. The animals were grouped and then dosed and challenged. 1 hour before each aerosol stimulation, the mice were anesthetized and fixed in a supine position on a 60° inclined surface, and intratracheal dry powder aerosolization was performed to administer lactose powder or compound BIOS-B12 (1.5mg / kg). On day 26, 24 hours after the last aerosol stimulation, the mice underwent a methacholine provocation test, and the airway responsiveness of the mice was tested using the animal respiratory function monitoring system (EMMS-WBP), and the penh value was recorded. The results are shown in Figure 1 (MCH is acetylcholine). On day 27, after mice were anesthetized, the neck skin was incised, the trachea was isolated, and PBS was perfused into the lungs through the trachea and aspirated to obtain bronchoalveolar lavage fluid (BALF). This was repeated twice, with 0.6 mL of BALF perfusion. The BALF was centrifuged at 1500 rpm for 10 minutes at 4°C. The cell pellet was resuspended in 1 mL of PBS, and the total number of cells in the mouse BALF was counted using a cell counter. The results are shown in Figure 2.

[0364] Ovalbumin can induce allergic asthma in mice. Model control animals exhibited significant airway hyperresponsiveness in response to methacholine stimulation. Penh values ​​showed that administration of the compound BIOS-B12 improved airway hyperresponsiveness in mice. Furthermore, administration of the compound BIOS-B12 significantly inhibited the accumulation of cells in bronchoalveolar lavage fluid following allergen stimulation.

[0365] Example 29. Pharmacokinetics of the compounds of the present invention after pulmonary administration

[0366] Male Sprague-Dawley rats were divided into a blank group and groups administered a single dose 15 minutes, 30 minutes, 1 hour, 2 hours, and 4 hours after dosing. Blood samples were collected at the corresponding time points, and plasma was separated. After blood collection, the animals were sacrificed, and lung tissue was obtained. Lung tissue homogenates were prepared and centrifuged. The supernatant was removed and the concentrations of B' (reference compound 2) and BIOS-B12 in the lung tissue of the Sprague-Dawley rats were determined by LC-MS / MS. Pharmacokinetic parameters were calculated, and the metabolic profiles of B' (reference compound 2) and BIOS-B12 in the lung tissue of the animals after dosing were investigated. The results are shown in Table 3.

[0367] Table 3: Pharmacokinetic data of the compounds of the present invention

[0368] As can be seen from Table 3, BIOS-B12 has a larger exposure amount than the control compound 2, which is more conducive to inhalation administration into the lungs.

[0369] Example 30. Pharmacokinetic study of the compounds of the present invention after pulmonary administration (measuring lung-blood ratio)

[0370] Male Sprague-Dawley rats were divided into a blank group and groups administered a single dose 15 minutes, 30 minutes, 1 hour, 2 hours, and 4 hours after administration. Blood samples were collected at the corresponding time points, and plasma was separated. After blood collection, the animals were sacrificed, and lung tissue was harvested. Lung tissue homogenates were prepared and centrifuged, and the supernatant was removed. The concentrations of B' (reference compound 2), BIOS-B12, BIOS-B5, and BIOS-A3 in the plasma and lung tissue of the Sprague-Dawley rats were determined by LC-MS / MS. Pharmacokinetic parameters in plasma and lung tissue were analyzed to calculate the lung-to-blood ratio. The metabolic profiles of B' (reference compound 2), BIOS-B12, BIOS-B5, and BIOS-A3 in the animals after administration were investigated.

[0371] The results are shown in Table 4.

[0372] Table 4: Lung-to-blood ratio data of the compounds of the present invention

[0373] As shown in Table 4, BIOS-B12, BIOS-B5, and BIOS-A3 have a larger lung-to-blood ratio than the control compound 2, which is more conducive to inhalation administration into the lungs.

[0374] Example 31. Inhibitory effect of compounds on hERG ion channels

[0375] The hERG inhibitory effect of some compounds of the present invention was tested using a traditional model to determine safety. The results are shown in Table 5.

[0376] Table 5: Blocking rate of compounds on hERG ion channels

[0377] The results showed that the compound BIOS-B12 of the present invention had no significant inhibitory effect on hERG channels.

Claims

1. An aromatic ring thiazine derivative, which is a compound represented by the general formula I or its isomers, or a pharmaceutically acceptable salt thereof: Where: W is selected from direct bond, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl or C 1-6 Alkylamine; Ring A is selected from a benzene ring or a 5-8 membered aromatic heterocycle substituted by at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 2-6 Alkynyl, C 2-6 Alkenyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 1-6 One or more of alkylamine, amino, cyano, hydroxyl, carboxyl, carbonyl, and keto; R1 and R2 are each independently selected from hydrogen, halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 Heterocyclic, substituted or unsubstituted C 1-6 Alkylamino, cyano, hydroxyl, carboxyl or carbonyl; the C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 3-6 Heterocyclic group, C 1-6 Alkylamino groups may be substituted with at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino; Or, R1 and R2 are connected to form a monocyclic or bicyclic structure; R3, R4, and R5 are each independently selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Alkylamino, amino or cyano groups; R6 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 2-6 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl, substituted or unsubstituted 5-8 membered heteroaryl, amino, cyano or hydroxyl; the C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl and 5-8 membered heteroaryl may be replaced by at least one R 11 Substitution, the R 11 Selected from or a substituted or unsubstituted 5-8 membered aromatic ring or aromatic heterocyclic ring, wherein the 5-8 membered aromatic ring or aromatic heterocyclic ring may be substituted by at least one of the following groups: hydrogen, halogen or C 1-6 alkyl; R7, R8, R9, R 10 Each independently selected from C 1-6 Alkyl or halogenated C 1-6 alkyl; n, p, q are each independently selected from 0, 1, 2, 3, 4, 5 or 6.

2. The aromatic ring thiazine derivative according to claim 1, characterized in that: It is a compound represented by general formula II or its isomers, or a pharmaceutically acceptable salt thereof: Where: W is selected from a direct bond or C 1-6 alkyl; Ring A is selected from a benzene ring, pyrazole or pyridine substituted with at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 1-6 Alkylamino, amino or cyano groups; R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 heterocyclic group or cyano group; said C 1-6 Alkyl, C 3-6 Cycloalkyl, C 3-6 The heterocyclic group may be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino; R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano; R6 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted 5-8 membered heteroaryl; the C 1-6 Alkyl and 5-8 membered heteroaryl groups may be replaced by at least one R 11 Substitution, the R 11 Selected from R9 is selected from C 1-6 Alkyl or halogenated C 1-6 alkyl; n, p, q are each independently selected from 0, 1, 2 or 3.

3. The aromatic ring thiazine derivative according to claim 2, characterized in that: In the general formula II, when ring A is a benzene ring substituted with at least one R0, R1 is selected from C 1-6 or, when ring A is pyrazole or pyridine substituted with at least one R0, R1 is selected from C 1-6 Alkyl or C 3-6 Cycloalkyl.

4. The aromatic ring thiazine derivative according to claim 2, characterized in that: In the general formula II, W is selected from C 1-6 alkyl.

5. The aromatic ring thiazine derivative according to claim 2, characterized in that: It is a compound represented by general formula II-1 or its isomers, or a pharmaceutically acceptable salt thereof: Where: Ring A is selected from a benzene ring substituted with at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 1-6 Alkylamino, amino or cyano groups; R1 is selected from halogen, substituted or unsubstituted C 2-6 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 heterocyclic group or cyano group; said C 2-6 Alkyl, C 3-6 Cycloalkyl, C 3-6 The heterocyclic group may be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino; R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano; R6 is selected from halogen, C 1-6 Alkyl or 5-8 membered heteroaryl.

6. The aromatic ring thiazine derivative according to claim 5, characterized in that: In the general formula II-1, ring A is selected from a benzene ring.

7. The aromatic ring thiazine derivative according to claim 5, characterized in that: In the general formula II-1, R1 is selected from substituted or unsubstituted C 3-6 Cycloalkyl, when C 3-6 When a cycloalkyl group has a substituent, it may be substituted by at least one of the following groups: halogen, C 1-6 Alkyl, hydroxy or amino.

8. The aromatic ring thiazine derivative according to claim 7, characterized in that: In the general formula II-1, R1 is selected from C 3-6 Cycloalkyl.

9. The aromatic ring thiazine derivative according to claim 5, characterized in that: In the general formula II-1, R1 is selected from substituted or unsubstituted C 2-6 Alkyl, when C 2-6 When an alkyl group has a substituent, it may be substituted by at least one of the following groups: halogen, hydroxyl or amino.

10. The aromatic ring thiazine derivative according to claim 2, characterized in that: It is a compound represented by general formula II-2 or its isomers, or a pharmaceutically acceptable salt thereof: Where: Ring A is selected from pyrazole or pyridine substituted with at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, C 1-6 Alkylamino, amino or cyano groups; R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 3-6 heterocyclic group or cyano group; said C 1-6 Alkyl, C 3-6 Cycloalkyl, C 3-6 The heterocyclic group may be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino; R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano; and R4 and R5 are not amino at the same time; R6 is selected from halogen, C 1-6 Alkyl or 5-8 membered heteroaryl.

11. The aromatic ring thiazine derivative according to claim 10, characterized in that: In the general formula II-2, ring A is selected from pyrazole or pyridine substituted by at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl.

12. The aromatic ring thiazine derivative according to claim 10, characterized in that: In the general formula II-2, R1 is selected from substituted or unsubstituted C 1-6 Alkyl, when C 1-6 When an alkyl group has a substituent, it may be substituted by at least one of the following groups: halogen, hydroxyl or amino.

13. The aromatic ring thiazine derivative according to claim 1, characterized in that: It is a compound represented by general formula III or its isomers, or a pharmaceutically acceptable salt thereof: Where: Ring A is selected from a benzene ring, pyrazole or pyridine substituted with at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or halogenated C 1-6 Alkoxy; R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl or substituted or unsubstituted C 3-6 Cycloalkyl; the C 1-6 Alkyl, C 3-6 The cycloalkyl groups may be substituted with at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino; R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano; R 11 Selected from or a substituted or unsubstituted 5-8 membered aromatic ring or aromatic heterocyclic ring; wherein the 5-8 membered aromatic ring or aromatic heterocyclic ring may be substituted by at least one of the following groups: hydrogen or C 1-6 alkyl; R7, R8, R9, R 10 Each independently selected from C 1-6 Alkyl or halogenated C 1-6 alkyl; n, p, q are each independently selected from 0, 1, 2 or 3.

14. The aromatic ring thiazine derivative according to claim 13, characterized in that: In the general formula III, ring A is selected from a benzene ring substituted by at least one R0, wherein R0 is selected from hydrogen, halogen or C 1-6 alkyl; R1 is selected from C 1-6 alkyl; R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano; R 11 Selected from 15. The aromatic ring thiazine derivative according to claim 1, characterized in that: It is a compound represented by general formula III or its isomers, or a pharmaceutically acceptable salt thereof: In the formula: Ring A is selected from a benzene ring, pyrazole or pyridine substituted by at least one R0, wherein R0 is selected from hydrogen, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or halogenated C 1-6 Alkoxy; R1 is selected from halogen, substituted or unsubstituted C 1-6 Alkyl or substituted or unsubstituted C 3-6 Cycloalkyl; the C 1-6 Alkyl, C 3-6 The cycloalkyl groups may be substituted with at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino; R3, R4, and R5 are each independently selected from C 1-6 Alkyl, amino or cyano; R 11 Selected from R9, R 10 Each independently selected from substituted C 1-6 Alkyl, substituted or unsubstituted 5-8 membered heterocyclic group, when C 1-6 When an alkyl group or a 5-8 membered heterocyclic group has a substituent, it may be substituted by at least one of the following groups: 1-6 Alkoxy; n is selected from 0, 1, 2 or 3.

16. The aromatic thiazine derivative according to any one of claims 1 to 15, characterized in that R3, R4, and R5 in the general formula I, general formula II, general formula II-1, general formula II-2, and general formula III can be further defined as: R3 is selected from cyano, R4 is selected from C 1-6 Alkyl, R5 is selected from amino.

17. The aromatic thiazine derivative according to any one of claims 1 to 15, characterized in that In the general formula I, general formula II, general formula II-1, general formula II-2 and general formula III, the heterocyclic group, heteroaryl group and aromatic heterocycle contain at least one heteroatom selected from N, O or S.

18. An aromatic ring thiazine derivative selected from the following characteristic compounds or their isomers, or pharmaceutically acceptable salts thereof:

19. A pharmaceutical composition comprising at least one compound as described in any one of Claims 1 to 18 and at least one pharmaceutically acceptable carrier or excipient.

20. Use of the compound according to any one of claims 1 to 18 or the pharmaceutical composition according to claim 19 in preventing or treating diseases associated with PI3Kδ kinase activity.

21. The use according to claim 20, characterized in that The drug is used for preventing or treating allergic diseases and inflammatory diseases; in particular, it plays a role in preventing or treating asthma, COPD and autoimmune diseases associated with PI3Kδ deficiency by delivery via inhalation administration.

22. The use according to claim 21, characterized in that The allergic diseases and inflammatory diseases are selected from asthma of any type or cause, including but not limited to intrinsic asthma, exogenous asthma, mild asthma, moderate asthma, severe asthma, bronchitis asthma, exercise-induced asthma, occupational asthma, Th2 asthma and non-Th2 asthma, wheezing infant syndrome, acute lung injury, chronic obstructive pulmonary disease, chronic bronchitis, emphysema, adult-onset / acute respiratory distress syndrome and other respiratory diseases; and autoimmune diseases such as rheumatoid arthritis, osteoarthritis, lupus erythematosus, psoriasis, allergic dermatitis, multiple sclerosis and the like.

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