Aromatic ring thiazine derivative and medical application thereof

Abstract

The invention belongs to the technical field of medicines, and relates to an aromatic ring thiazine derivative, in particular to structural forms such as free alkali, isomeride and pharmaceutically acceptable salt form of the compound. Methods for preparing such compounds; compositions comprising such compounds and therapeutic uses thereof. Based on a histamine PI3K delta receptor ligand, a series of compounds with novel structures are developed, a series of related biological tests are carried out on the compounds, and test results show that the compounds find novel PI3K delta high-selectivity inhibitors, have the characteristics of small toxic and side effects and low bioavailability, and have broad application prospects. The pharmaceutical composition is used for developing inhaled medicines suitable for treating respiratory system diseases such as asthma and COPD, and has important clinical significance and social value.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and relates to an aromatic ring-fused thiazine derivative, specifically to structural forms such as free bases, isomers, and pharmaceutically acceptable salt forms of such compounds; methods for preparing such compounds; compositions containing such compounds and their therapeutic uses. Background Art

[0002] The PI3K / Akt / mTOR pathway is a signal transduction pathway that is abnormally activated in human cancers and is closely related to the occurrence and development of malignant tumors. As a key kinase in this signal pathway, a variety of PI3K inhibitors have entered clinical research successively, and it is a popular target for the research and development of anti-tumor drugs.

[0003] PI3K signaling is one of the most frequently abnormally activated pathways. Early studies have shown that the pan-PI3K inhibitors LY294002 and wortmannin can reverse the drug resistance of cancer cells to various therapies. At the same time, some PI3K family members are also involved in inflammation and autoimmune functions. Class I PI3Ks consist of complex regulatory subunits and carry the p110 catalytic subunit (p110α, β, γ or δ). These heterodimeric complexes are called PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ, and p110α, p110β, p110γ, and p110δ represent the catalytic subunits themselves. p110α and p110β show a wide tissue distribution, and the p110γ and p110δ isoforms are mainly 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. Therefore, the PI3Kδ and PI3Kγ isoforms are extremely relevant to inflammatory respiratory diseases.

[0004] Currently, only 4 varieties of PI3Kδ inhibitors have been marketed, including Idelalisib, Copanlisib, Duvelisib, and Umbralisib. Among them, 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 the Chinese counterpart CN101031569B, protects the structure of the PI3Kδ inhibitor Idelalisib (CAL-101, Zydelig). In July 2014, Idelalisib obtained the approval of the US FDA 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 of idelalisib carries a black box warning about possible side effects such as liver toxicity, pneumonia, severe diarrhea, enteritis, and intestinal perforation induced by idelalisib.

[0006] Patent WO2011008302A1, corresponding to the Chinese counterpart CN102711767B, protects the PI3Kδ / γ selective inhibitor Duvelisib (IPI-145, INK-1197). In September 2018, Duvelisib obtained the approval of the US FDA for the treatment of adult patients with relapsed or refractory chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL) who have received at least 2 prior therapies and for the treatment of adult patients with relapsed or refractory follicular lymphoma (FL) who have received at least 2 prior therapies. It should be noted that the drug label of Duvelisib contains a black box warning indicating fatal and serious toxic effects, including infections, diarrhea or colitis, skin reactions, and pneumonia.

[0007] Based on the defects of the toxic and side effects of the above two PI3Kδ inhibitors, domestic and foreign scientific researchers have continued to modify the compound structure on their basis, hoping to find a safer and more effective PI3Kδ inhibitor.

[0008] Patent US20130053362A1, corresponding to the Chinese counterpart CN103998442A, discloses a compound and a pharmaceutical composition for inhibiting phosphoinositide 3-kinase (PI3K). The parent nucleus of the compound has the structures of isoquinolinone and quinazolinone and can be used for the treatment of cancer, inflammatory diseases, or autoimmune diseases.

[0009] Patent CN107033145B protects a kind of benzothiazine and benzothiadiazine compounds. Most of these compounds selectively have a significant inhibitory effect on PI3Kδ and can be used for the preparation of anti-inflammatory and anti-tumor drugs. The representative compound S-I-11 has an IC 50It is 0 - 20 nM. As known from the literature (Eur. J. Med. Chem. 2019, 170, 112 - 125), the oral bioavailability of this molecule (S - 63 in the literature) is 29.2%.

[0010] In summary, the cores of the above PI3K inhibitors are mainly isoquinolinone and quinazolinone, and they are all used to prepare drugs for treating anti - tumor, anti - inflammatory, and immune diseases. Patent CN107033145B is based on Idelalisib and Duvelisib and designed compounds with thiazine and benzothiadiazine as the skeletons using the principle of bioisosteres, which are also used for the treatment of tumor diseases and are suitable for oral administration. Thus, in the field of PI3Kδ inhibitors, the research on structural types with high PI3Kδ selectivity and using inhalation administration for treating respiratory diseases such as asthma and COPD is still scarce.

[0011] Therefore, it is urgent for those skilled in the art to search for and discover novel highly selective PI3Kδ inhibitors, which have the characteristics of low toxicity and side effects and low bioavailability, for developing inhalation drugs suitable for treating respiratory diseases such as asthma and COPD, having important clinical significance and social value. Summary of the Invention

[0012] The present invention aims to provide a novel aromatic ring - fused thiazine derivative, which is a highly selective PI3Kδ inhibitor with an oral bioavailability of 0 and a fast clearance rate, and can be used to develop inhalation preparations for treating respiratory diseases such as asthma and COPD.

[0013] To solve the above - mentioned technical problems, the technical solutions adopted by the present invention are as follows specifically:

[0014] The present invention provides an aromatic ring - fused thiazine derivative, which is a compound shown in general formula I or its isomers, or its pharmaceutically acceptable salts:

[0015]

[0016] In the formula:

[0017] W is selected from a direct bond, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl or C 1-6 alkylamino;

[0018] Ring A is selected from a benzene ring substituted by at least one R 0 or a 5 - 8 - membered heteroaromatic ring, and R 0 is selected from hydrogen, halogen, C 1-6 alkyl, C 2-6 alkynyl, C 2-6Alkenyl, C 1-6 Alkoxy, halo C 1-6 Alkyl, halo C 1-6 Alkoxy, C 1-6 Alkylamino, amino, cyano, hydroxy, carboxyl, carbonyl, ketone group, or one or more of them;

[0019] R 1 、R 2 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, hydroxy, carboxyl or carbonyl; C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 3-6 Heterocyclic group, C 1-6 Alkylamino can be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 Alkyl, hydroxy or amino;

[0020] Or, R 1 Is connected to R 2 To form a monocyclic or bicyclic structure;

[0021] R 3 、R 4 、R 5 Each independently selected from C 1-6 Alkyl, halo C 1-6 Alkyl, C 3-6 Cycloalkyl, C 1-6 Alkylamino, amino or cyano;

[0022] R 6 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- to 8-membered heteroaryl, amino, cyano or hydroxy; C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, 5- to 8-membered heteroaryl can be substituted by at least one R 11 Substituted, R 11 Selected from Or substituted or unsubstituted 5- to 8-membered aromatic ring or heteroaromatic ring, where the 5- to 8-membered aromatic ring or heteroaromatic ring can be substituted by at least one of the following groups: hydrogen, halogen or C 1-6 Alkyl;

[0023] R 7 、R 8 、R 9 、R 10 are each independently selected from C 1-6 alkyl or halo C 1-6 alkyl;

[0024] n, p, and q are each independently selected from 0, 1, 2, 3, 4, 5, or 6.

[0025] The present invention also provides an aryl-fused thiazine derivative, which may be a compound represented by General Formula II or its isomers, or its pharmaceutically acceptable salts:

[0026]

[0027] In the formula:

[0028] W is selected from a direct bond or C 1-6 alkyl;

[0029] Ring A is selected from a benzene ring, pyrazole, or pyridine that is at least substituted by one R 0 , R 0 is selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkyl, halo C 1-6 alkoxy, C 1-6 alkylamino, amino, or cyano;

[0030] R 1 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; C 1-6 alkyl, C 3-6 cycloalkyl, C 3-6 heterocyclic group may each be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 alkyl, hydroxy, or amino;

[0031] R 3 、R 4 、R 5 are each independently selected from C 1-6 alkyl, amino, or cyano;

[0032] R 6 is selected from halogen, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted 5- to 8-membered heteroaryl; C 1-6 alkyl, 5- to 8-membered heteroaryl may each be substituted by at least one R 11 substituted, R11 Selected from

[0033] R 9 Selected from C 1-6 alkyl or halo C 1-6 alkyl;

[0034] n, p, and q are each independently selected from 0, 1, 2, or 3.

[0035] Preferably, in the foregoing general formula II, when ring A is a benzene ring substituted with at least one R 0 substituent, R 1 is selected from C 1-6 alkyl; or, when ring A is a pyrazole or pyridine substituted with at least one R 0 substituent, R 1 is selected from C 1-6 alkyl or C 3-6 cycloalkyl.

[0036] Preferably, in the foregoing general formula II, W is selected from C 1-6 alkyl.

[0037] Preferably, in the foregoing general formula II, W is selected from C 1-6 alkyl, ring A is selected from a benzene ring, pyrazole, or pyridine, R 1 is selected from C 1-6 alkyl, R 3 is selected from cyano, R 4 is selected from C 1-6 alkyl, R 5 is selected from amino, R 6 is selected from halogen.

[0038] The present invention also provides an aromatic ring-fused thiazine derivative, which may be a compound represented by general formula II-1 or its isomers, or its pharmaceutically acceptable salts:

[0039]

[0040] In the formula:

[0041] Ring A is selected from a benzene ring substituted with at least one R 0 substituent, and the said R 0 is selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkyl, halo C 1-6 alkoxy, C 1-6 alkylamino, amino, or cyano;

[0042] R 1 is selected from halogen, substituted or unsubstituted C 2-6 alkyl, substituted or unsubstituted C 3-6Cycloalkyl, substituted or unsubstituted C 3-6 heterocyclic group or cyano group; C 2-6 alkyl group, C 3-6 cycloalkyl group, C 3-6 heterocyclic group can each be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 alkyl group, hydroxyl group or amino group;

[0043] R 3 、R 4 、R 5 are each independently selected from C 1-6 alkyl group, amino group or cyano group;

[0044] R 6 is selected from halogen, C 1-6 alkyl group or 5- to 8-membered heteroaryl group.

[0045] Preferably, in the aforementioned general formula II-1, ring A is selected from a benzene ring.

[0046] Preferably, in the aforementioned general formula II-1, R 1 is selected from substituted or unsubstituted C 3-6 cycloalkyl group. When there are substituents on the C 3-6 cycloalkyl group, it can be substituted by at least one of the following groups: halogen, C 1-6 alkyl group, hydroxyl group or amino group.

[0047] Preferably, in the aforementioned general formula II-1, R 1 is selected from C 3-6 cycloalkyl group.

[0048] Preferably, in the aforementioned general formula II-1, R 1 is selected from substituted or unsubstituted C 2-6 alkyl group. When there are substituents on the C 2-6 alkyl group, it can be substituted by at least one of the following groups: halogen, hydroxyl group or amino group.

[0049] Preferably, in the aforementioned general formula II-1, R 3 is selected from cyano group.

[0050] Preferably, in the aforementioned general formula II-1, R 4 is selected from C 1-6 alkyl group.

[0051] Preferably, in the aforementioned general formula II-1, R 5 is selected from amino group.

[0052] Preferably, in the aforementioned general formula II-1, R 6 is selected from halogen or C 1-6 alkyl group.

[0053] Preferably, in the aforementioned general formula II-1, ring A is selected from a benzene ring, R1 Selected from C 3-6 cycloalkyl, R 3 Selected from cyano, R 4 Selected from C 1-6 alkyl, R 5 Selected from amino, R 6 Selected from halogen.

[0054] The present invention also provides an aromatic ring-fused thiazine derivative, which can be a compound represented by the general formula II-2 or its isomer, or its pharmaceutically acceptable salt:

[0055]

[0056] In the formula:

[0057] Ring A is selected from pyrazole or pyridine substituted by at least one R 0 , and the R 0 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;

[0058] R 1 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; C 1-6 alkyl, C 3-6 cycloalkyl, C 3-6 heterocyclic group can be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 alkyl, hydroxyl or amino;

[0059] R 3 , R 4 , R 5 are each independently selected from C 1-6 alkyl, amino or cyano; and R 4 , R 5 are not both amino at the same time;

[0060] R 6 is selected from halogen, C 1-6 alkyl or 5-8 membered heteroaryl.

[0061] Preferably, in the aforementioned general formula II-2, ring A is selected from pyrazole or pyridine substituted by at least one R 0 , and R 0 is selected from hydrogen, halogen, C 1-6 alkyl or halogenated C 1-6 alkyl.

[0062] Preferably, in the aforementioned general formula II-2, R 1 is selected from substituted or unsubstituted C 1-6 alkyl. When the C 1-6 alkyl has substituents, it may be substituted by at least one of the following groups: halogen, hydroxyl, or amino.

[0063] Preferably, in the aforementioned general formula II-2, R 3 is selected from cyano.

[0064] Preferably, in the aforementioned general formula II-2, R 4 is selected from C 1-6 alkyl.

[0065] Preferably, in the aforementioned general formula II-2, R 5 is selected from amino.

[0066] Preferably, in the aforementioned general formula II-2, R 6 is selected from halogen or C 1-6 alkyl.

[0067] Preferably, in the aforementioned general formula II-2, ring A is selected from pyrazole, pyridine, or pyridine substituted by at least one R 0 wherein R 0 is selected from halogen, R 1 is selected from C 1-6 alkyl, R 3 is selected from cyano, R 4 is selected from C 1-6 alkyl, R 5 is selected from amino, R 6 is selected from halogen.

[0068] The present invention also provides an aromatic ring-fused thiazine derivative, which can be a compound represented by the general formula III, its isomers, or its pharmaceutically acceptable salts:

[0069]

[0070] In the formula:

[0071] Ring A is selected from a benzene ring, pyrazole, or pyridine substituted by at least one R 0 wherein R 0 is selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, halogenated C 1-6 alkyl, or halogenated C 1-6 alkoxy;

[0072] R 1 is selected from halogen, substituted or unsubstituted C 1-6 alkyl, or substituted or unsubstituted C 3-6 cycloalkyl; C1-6 Alkyl, C 3-6 Cycloalkyl may each be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 alkyl, hydroxyl or amino;

[0073] R 3 、R 4 、R 5 are each independently selected from C 1-6 alkyl, amino or cyano;

[0074] R 11 is selected from or a substituted or unsubstituted 5- to 8-membered aromatic or heteroaromatic ring; wherein the 5- to 8-membered aromatic or heteroaromatic ring may be substituted by at least one of the following groups: hydrogen or C 1-6 alkyl;

[0075] R 7 、R 8 、R 9 、R 10 are each independently selected from C 1-6 alkyl or halo C 1-6 alkyl;

[0076] n, p, q are each independently selected from 0, 1, 2 or 3.

[0077] The present invention also provides an aromatic ring-fused thiazine derivative, which may be a compound represented by the general formula III or its isomers, or its pharmaceutically acceptable salts:

[0078]

[0079] In the formula:

[0080] Ring A is selected from a benzene ring, pyrazole or pyridine which is substituted by at least one R 0 , and R 0 is selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 alkoxy, halo C 1-6 alkyl or halo C 1-6 alkoxy;

[0081] R 1 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 cycloalkyl may each be substituted by at least one of the following groups: hydrogen, halogen, C 1-6 alkyl, hydroxyl or amino;

[0082] R 3 、R 4 、R5 Each independently selected from C 1-6 alkyl, amino or cyano;

[0083] R 11 Selected from

[0084] R 9 、R 10 Each independently selected from substituted C 1-6 alkyl, substituted or unsubstituted 5- to 8-membered heterocyclic group. When there are substituents on the C 1-6 alkyl or 5- to 8-membered heterocyclic group, they can be substituted by at least one of the following groups: C 1-6 alkoxy;

[0085] n is selected from 0, 1, 2 or 3.

[0086] In the aforementioned general formula III, ring A is selected from a benzene ring substituted by at least one R 0 , R 0 is selected from hydrogen, halogen or C 1-6 alkyl; R 1 is selected from C 1-6 alkyl; R 3 、R 4 、R 5 Each independently selected from C 1-6 alkyl, amino or cyano;

[0087] R 11 Selected from

[0088] Preferably, in the aforementioned general formula I, general formula II, general formula II-1, general formula II-2, general formula III, R 3 、R 4 、R 5 can be further defined as: R 3 is selected from cyano, R 4 is selected from C 1-6 alkyl, R 5 is selected from amino.

[0089] Preferably, in the aforementioned aromatic ring-fused thiazine derivatives, the heterocyclic group, heteroaryl group, and arylheterocyclic group contain at least one heteroatom, and the heteroatom is selected from N, O or S.

[0090] More preferably, the arylheterocyclic group can include but is not limited to those selected from furan, thiophene, pyrazole, pyridine, pyrimidine, etc.

[0091] More preferably, the heteroaryl group can include but is not limited to those selected from furyl, thienyl, pyrazolyl, pyridyl, pyrimidinyl, etc. More preferably, the heterocyclic group can include but is not limited to those selected from pyrrole, morpholine, piperidine, piperazine, etc.

[0092] The present invention also provides an aromatic ring-fused thiazine derivative, including the following compounds numbered BIOS-A1 to BIOS-A8, BIOS-B1 to BIOS-B17 or pharmaceutically acceptable salts thereof:

[0093] 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;

[0094] 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;

[0095] 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;

[0096] 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;

[0097] 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;

[0098] 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;

[0099] 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;

[0100] 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;

[0101] 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;

[0102] 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;

[0103] 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;

[0104] 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;

[0105] 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;

[0106] 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]heptan-5-yl)-6-methylpyrimidine-5-carbonitrile;

[0107] 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;

[0108] 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;

[0109] 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;

[0110] 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;

[0111] 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;

[0112] 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;

[0113] 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;

[0114] 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;

[0115] BIOS - B16: (S)-2 - Amino - 4 - ((1-(1,1 - dihydroxy - 8-(6 - oxo - 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;

[0116] 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;

[0117] The compounds numbered BIOS-A1 to BIOS-A8, BIOS-B1 to BIOS-B17 above or their pharmaceutically acceptable salts have the following specific structural formulas:

[0118]

[0119]

[0120] The present invention also provides a method for preparing the aforementioned compounds, and the steps are as follows (including but not limited to the following methods):

[0121] Route 1: Preparation of Series A Compounds

[0122]

[0123] The specific reaction process is as follows:

[0124] The o-halogenated aromatic amine compound A-001 reacts with hydrochloric acid, sodium nitrite, sodium sulfite and copper sulfate to form a sulfonyl chloride intermediate A-002; then it undergoes a condensation reaction with an amino compound to form an intermediate A-003; the chiral (S)-2-(but-3-yn-2-yl)isoindoline-1,3-dione A-004 and the intermediate A-003 are catalyzed by a metal catalyst and undergo a coupling ring-closure reaction to obtain an intermediate A-005; the intermediate A-005 and hydrazine hydrate are heated under reflux in an ethanol solvent, and a deprotection reaction is carried out to obtain an intermediate A-006, and the latter undergoes an SN2 reaction with a substituted aminopyrimidine to obtain a series of target compounds, including but not limited to the compounds BIOS-A1 to BIOS-A7.

[0125] In addition, the product BIOS-A6 prepared by this reaction route can be further reacted with an alkyne derivative in one step under alkaline conditions with a noble metal as a catalyst to obtain the compound BIOS-A8.

[0126] Route 2: Preparation of Series B Compounds

[0127]

[0128] The specific reaction process is as follows:

[0129] The o-halogenated aromatic amine compound B-001 reacts with hydrochloric acid, sodium nitrite, sodium sulfite and copper sulfate to form a sulfonyl chloride intermediate B-002, which is then condensed with an amino compound to form an intermediate B-003; the Boc-protected chiral amino acid intermediate B-004 reacts with the intermediate B-003 at low temperature to obtain an intermediate B-005; the intermediate B-005 is reacted in concentrated hydrochloric acid to obtain B-006, and the latter undergoes an SN2 reaction with a substituted aminopyrimidine to obtain a series of target compounds, including but not limited to the compounds BIOS-B1 to BIOS-B10.

[0130] In addition, the products prepared by this reaction route can be further reacted with alkyne derivatives in one step using noble metals as catalysts under alkaline conditions to obtain compounds BIOS-B11 to BIOS-B17.

[0131] The "compounds" described in the present invention include, but are not limited to, the following forms of compounds: free bases, stereoisomers, geometric isomers, tautomers, isotopes, pharmaceutically acceptable salts, solvates, hydrates, prodrugs (esters or phosphates), etc.

[0132] The "compounds" described in the present invention can be asymmetric, for example, having one or more stereoisomers. Unless otherwise specified, all stereoisomers are included, such as enantiomers and diastereomers. Compounds containing asymmetric carbon atoms in the present invention can be isolated in optically active pure form or racemic form. The optically active pure form can be obtained by methods such as resolution of racemic mixtures, synthesis using chiral starting materials or chiral reagents.

[0133] The "isomers" described in the present invention refer to, unless otherwise specified, stereoisomers or tautomers. Unless otherwise specified, the term "stereoisomer" refers to a compound having the same chemical structure but different arrangements of atoms or groups in space. Stereoisomers include, but are not limited to, enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans isomers), atropisomers. Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, diastereomers based on differences in the physical and chemical properties of the components, for example, by chromatography and / or fractional crystallization. Unless otherwise specified, the term "tautomer" refers to structural isomers that can be interconverted through a low energy barrier with different energies. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also called proton transfer tautomers) include interconversions through proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions through recombination of some bonding electrons.

[0134] As used herein, the term "isotope" means that, unless otherwise specified, the compounds of the present invention may exist in isotopically labeled or enriched forms, containing one or more atoms whose atomic weights or mass numbers are different from those of the most abundant atoms found in nature. Isotopes can be radioactive or non-radioactive. Isotopes commonly used for isotope labeling 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 18 O; sulfur isotopes: including but not limited to 35 S. These isotopically labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues, especially 3 H and 13 C, which are more widely used because they are easy to label and convenient to detect. Substitution with certain heavy isotopes, such as deuterium ( 2 H), can enhance metabolic stability, extend the half-life, and thus provide a therapeutic advantage by reducing the dose. Isotopically labeled compounds are generally synthesized starting from labeled starting materials using known synthetic techniques in the same way as non-isotopically labeled compounds.

[0135] As used herein, the term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from compounds having specific substituents found in the present invention and relatively non-toxic bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds 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. For example: alkali metal salts, alkaline earth metal salts, other metal salts, inorganic base salts, organic base salts, inorganic acid salts, lower alkanesulfonates, arylsulfonates, organic acid salts, amino acid salts, etc.

[0136] In addition to the salt form, the compounds provided by the present invention also exist in the form of prodrugs. The prodrugs of the compounds described herein are readily chemically changed under physiological conditions to convert to the compounds of the present invention. In addition, the prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in the in vivo environment.

[0137] For the compounds containing the foregoing general formula structure, the terms used herein have the following meanings:

[0138] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0139] The term "cyano" refers to -CN.

[0140] The term "amino" refers to -NH 2 .

[0141] The term "alkyl" refers to a straight-chain or branched-chain saturated hydrocarbon group composed of carbon atoms and hydrogen atoms, such as C 1-6 alkyl, including but not limited to C 2-6 alkyl, C 3-6 alkyl, etc. Non-limiting examples thereof 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 for another example, C 2-6 alkyl means the case where methyl is excluded from C 1-6 alkyl.

[0142] The term "alkenyl" refers to a straight-chain or branched-chain unsaturated aliphatic hydrocarbon group composed of carbon atoms and hydrogen atoms and having at least one double bond. The alkenyl can contain 2-20 carbon atoms, preferably 2-10 carbon atoms (i.e., C 2-10 alkenyl), more preferably 2-8 carbon atoms (i.e., C 2-8 alkenyl), still more preferably 2-6 carbon atoms (i.e., C 2-6 alkenyl), 2-5 carbon atoms (i.e., C 2-5 alkenyl), 2-4 carbon atoms (i.e., C 2-4 alkenyl), 2-3 carbon atoms (i.e., C 2-3 alkenyl), 2 carbon atoms (i.e., C 2 alkenyl). For example, "C 2-6"Alkenyl" means that the group is alkenyl 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 include, but are not limited to, vinyl, 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, etc.

[0143] The term "alkynyl" refers to a straight-chain or branched-chain unsaturated aliphatic hydrocarbon group composed 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), more preferably 2 to 8 carbon atoms (C 2-8 alkynyl), still more preferably 2 to 6 carbon atoms (i.e., C 2-6 alkynyl), 2 to 5 carbon atoms (i.e., C 2-5 alkynyl), 2 to 4 carbon atoms (i.e., C 2-4 alkynyl), 2 to 3 carbon atoms (i.e., C 2-3 alkynyl), 2 carbon atoms (i.e., C 2 alkynyl). For example, "C 2-6 alkynyl" means that the group is alkynyl 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 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, etc.

[0144] The term "haloalkyl" refers to an alkyl group in which one or more hydrogens are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine, and iodine). Examples of haloalkyl include halo-C 1-8 alkyl, halo-C 1-6 alkyl, or halo-C 1-4 alkyl, but are not limited to -CF 3 , -CH 2 Cl, -CH 2 CF 3 , -CCl 2 , CF 3 , etc.

[0145] The term "C 1-6"Alkoxy" refers to an -O-alkyl group, where the alkyl group 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.

[0146] The term "C 1-6 alkylamino" refers to a C 1-6 monoalkyl-substituted amino group, or a di-C 1-6 alkyl-substituted amino group, where the alkyl group is as defined above. Examples of "C 1-6 monoalkylamino" include, but are not limited to, methylamino, ethylamino, propylamino, cyclopropylamino, isopropylamino; examples of "C 1-6 dialkylamino" include, but are not limited to, dimethylamino, diethylamino, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl.

[0147] The term "aromatic ring" refers to a fully carbonaceous monocyclic or fused polycyclic ring having 6 to 14 carbon atoms and a fully conjugated π-electron system, including, but not limited to, benzene ring, naphthalene ring, anthracene ring, etc., preferably a benzene ring.

[0148] The term "aryl" refers to a fully carbonaceous monocyclic or fused polycyclic group having 6 to 14 carbon atoms and a fully conjugated π-electron system, including, but not limited to, phenyl, naphthyl, anthryl, etc., preferably a phenyl group.

[0149] The term "heterocyclic ring" refers to a saturated or partially unsaturated monocyclic or polycyclic ring (such as a spiro ring, a bridged ring, etc.) containing 3 to 10 ring atoms, which is a non-aromatic structure; the polycyclic ring can be all non-aromatic rings, or at least one ring is an aromatic ring and the remaining rings are non-aromatic rings. One or more (such as 2, 3, 4 or more) of the aforementioned 3 to 10 ring atoms are heteroatoms, and the rest are carbon atoms. 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 can be one or more. Non-limiting examples of "heterocyclic ring" include, but are not limited to, aziridine, oxirane, thiirane, azetidine, oxetane, thietane, tetrahydrofuran, piperidine, piperazine, morpholine, thiomorpholine, etc.

[0150] The term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic or polycyclic (such as spiro, bridged, etc.) group containing 3 to 10 ring atoms, which is a non-aromatic structure; the polycycle can have all non-aromatic rings or at least one aromatic ring and the remaining non-aromatic rings. One or more (such as 2, 3, 4 or more) of the aforementioned 3 to 10 ring atoms are heteroatoms, and the rest are carbon atoms. The heteroatom is selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic can include the same or different heteroatoms in one or more rings, and the number of heteroatoms can be one or more. Non-limiting examples of "heterocyclic group" include, but are not limited to, aziridinyl, oxiranyl, thiiranyl, azetidinyl, oxetanyl, thietanyl, furyl, piperidinyl, piperazinyl, morpholinyl, pyrrolyl, thiomorpholinyl, etc.

[0151] The term "heteroaromatic ring" refers to an aromatic monocyclic or polycyclic (such as a fused ring, etc.) containing 5 to 14 ring atoms. One or more (such as 2, 3, 4 or more) of the aforementioned 5 to 14 ring atoms are heteroatoms, and the rest are carbon atoms. The heteroatom is selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic can include the same or different heteroatoms in one or more rings, and the number of heteroatoms can be one or more. The aforementioned "heteroaromatic ring" preferably contains 5 to 14, 5 to 10, 5 to 8 ring atoms, and more preferably contains 5 to 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, etc.

[0152] The term "heteroaryl" refers to an aromatic monocyclic or polycyclic (such as a fused ring, etc.) group containing 5 to 14 ring atoms. One or more (such as 2, 3, 4 or more) of the aforementioned 5 to 14 ring atoms are heteroatoms, and the rest are carbon atoms. The heteroatom is selected from one or more of N, O, and S. The aforementioned monocyclic or polycyclic can include the same or different heteroatoms in one or more rings, and the number of heteroatoms can be one or more. The aforementioned "heteroaryl" preferably contains 5 to 14, 5 to 12, 5 to 10, 5 to 8 ring atoms, and more preferably contains 5 to 6 ring atoms. Non-limiting examples of "heteroaryl" include, but are not limited to, furyl, thienyl, oxazolyl, thiazolyl, pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolinyl, indolyl, benzofuranyl, benzothienyl, benzimidazolyl, benzopyridinyl, benzopyrimidinyl, benzopyrazinyl, etc.

[0153] The present invention also provides a pharmaceutical composition, comprising at least one of the aforementioned compounds or a pharmaceutically acceptable salt thereof as an active ingredient, and at least one or more pharmaceutically acceptable carriers.

[0154] The "pharmaceutical composition" as described in the present invention refers to a preparation of one or more compounds or salts thereof of the present invention and a carrier commonly accepted in the art for delivering bioactive compounds to an organism (such as a human). The purpose of the pharmaceutical composition is to facilitate the administration and delivery to the organism.

[0155] In particular, the pharmaceutical composition provided by the present invention adopts an inhalable drug delivery mode.

[0156] The "inhalable" as described in the present invention means that such compounds can be administered through the airway and lungs, enabling absorption through lung epithelial cells or respiratory mucosa and directly entering the blood circulation. "Inhalable" indicates that it can be administered by inhalation, but is not limited to inhalation, and it is still applicable to other administration routes 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 preparations can be prepared by any method known in the pharmaceutical field. For example, by mixing the active ingredient with a carrier or excipient.

[0157] The present invention also provides the use of the aforementioned compounds or pharmaceutical compositions in the prevention or treatment of diseases related to PI3Kδ kinase activity.

[0158] Preferably, the aforementioned drug is used for the prevention or treatment of allergic diseases and inflammatory diseases; in particular, it plays a role in the prevention or treatment of asthma, COPD, and autoimmune diseases related to PI3Kδ deficiency through the delivery mode of inhalation administration.

[0159] More preferably, the aforementioned allergic diseases and inflammatory diseases are selected from asthma of any type or cause, including but not limited to endogenous asthma, exogenous asthma, mild asthma, moderate asthma, severe asthma, bronchitic asthma, exercise-induced asthma, occupational asthma, Th2-type asthma and non-Th2-type asthma, wheezy infant syndrome, acute lung injury, chronic obstructive pulmonary disease, chronic bronchitis, emphysema, adult / acute respiratory distress syndrome and other respiratory system diseases; and autoimmune diseases such as rheumatoid arthritis, osteoarthritis, lupus erythematosus, psoriasis, allergic dermatitis, multiple sclerosis, etc.

[0160] In particular, other inflammatory or obstructive airway diseases applicable to the present invention are selected from pneumoconiosis of any type or cause, including but not limited to asbestosis, anthracosis, siderosis, byssinosis, and cotton dust lung, etc.

[0161] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0162] Based on the target design of PI3Kδ inhibitors and using the principle of bioisosterism, the present invention has developed a series of structurally novel aromatic ring-fused thiazine derivatives that can be used for inhalation administration, and has conducted relevant biological tests. 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, when delivered by inhalation, some of the compounds can be well retained in the lungs, have good inhalation pharmacokinetic properties, and can be used as inhalation drugs for preventing or treating respiratory diseases such as asthma and COPD.

[0163] Description of the Drawings

[0164] Figure 1 It is a diagram showing the effect of compound BIOS-B12 on ovalbumin-induced airway hyperresponsiveness in mice.

[0165] Figure 2 It is a diagram showing the effect of compound BIOS-B12 on cell accumulation in the airways of ovalbumin-induced mice. Detailed Description of the Invention

[0166] The following are specific examples of the present invention, which further describe the technical solutions of the present invention, but the protection scope of the present invention is not limited to these examples. Any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the protection scope of the present invention.

[0167] 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:

[0168] Boc 2 O: Di-tert-butyl carbonate;

[0169] DMAP: 4-Dimethylaminopyridine;

[0170] Pd(PPh 3 ) 4 : Tetrakis(triphenylphosphine)palladium;

[0171] PPh 3 : Triphenylphosphine;

[0172] DIPEA: Diisopropylethylamine;

[0173] DEAD: Diethyl azodicarboxylate;

[0174] DMF: N,N-Dimethylformamide;

[0175] EA: Ethyl acetate;

[0176] PdCl 2 (CH3CN) 2 : Bis(acetonitrile)palladium(II) chloride;

[0177] Pd(amphos) 2 Cl 2 : Dichloro-di-tert-butyl-(4-dimethylaminophenyl)phosphine palladium(II);

[0178] HATU: 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate;

[0179] DMSO: Dimethyl sulfoxide;

[0180] X-phos: 2-Dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl.

[0181] In addition, all operations involving raw materials that are prone to oxidation or hydrolysis 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.

[0182] Each reaction starting material and common intermediate involved in the examples of the present invention can be obtained commercially or prepared by oneself. The starting materials and common intermediates that need to be prepared by oneself are described in detail in the preparation process as follows:

[0183] 1. Synthesis of intermediate 2-bromo-6-chlorobenzenesulfonyl chloride (BIOS-A1-1):

[0184]

[0185] Under an ice bath, 2-bromo-6-chloroaniline (7 g, 35 mmol) was added to concentrated hydrochloric acid (30 mL), and 7 mL of sodium nitrite solution (2.8 g, 41 mmol) was slowly added dropwise. In another reaction flask, copper sulfate pentahydrate (0.93 g, 3.7 mmol) and 32 mL of concentrated hydrochloric acid were added. Under an ice bath, 20 mL of sodium bisulfite solution (7.05 g, 67.8 mmol) and the above solution were slowly added dropwise. After the addition was completed, the reaction mixture was warmed to room temperature and reacted for 2 h until the raw materials were completely reacted. The reaction solution was extracted with dichloromethane (80 mL * 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product BIOS-A1-1 (7.0 g, 24.13 mmol). The product was directly used in the next step without purification.

[0186] 2. Synthesis of intermediate tert-butyl 4-amino-1H-pyrazole-1-carboxylate BIOS-A1-2:

[0187]

[0188] 4-Nitro-1H-pyrazole (1.1 g, 10 mmol) was dissolved in dichloromethane, and Boc was added respectively 2O (2.2 g, 10 mmol) and DMAP (110 mg, 1 mmol) were stirred at room temperature for 2 h. After the reaction was completed, the reaction was quenched by adding ammonium chloride solution. After concentration of the organic phase, tert-butyl 4-nitro-1H-pyrazole-1-carboxylate (1.7 g, 8 mmol) was obtained by purification through flash column chromatography. This intermediate was dissolved in methanol, nitrogen was charged, palladium on carbon was added, and a hydrogen balloon was attached. The mixture was purged with hydrogen three times and reacted overnight. After the reaction was completed, it was filtered through diatomaceous earth, and the filtrate was concentrated to obtain a pale yellow solid BIOS-A1-2 (0.9 g, 5 mmol).

[0189] 3. Synthesis of intermediate (S)-2-(but-3-yn-2-yl)isoindoline-1,3-dione A-004:

[0190]

[0191] Phthalimide (5.0 g, 34.3 mmol), (R)-3-butyn-2-ol (2.0 g, 28.6 mmol), and PPh 3 (9.0 g, 34.3 mmol) were dissolved in 50 mL of anhydrous tetrahydrofuran. DEAD (6.0 g, 34.3 mmol) was placed in an addition funnel with a constant pressure, and DEAD was added dropwise at 0 °C. After the addition was completed, the mixture was stirred at room temperature for 27 h. After the raw materials disappeared, the solvent was evaporated and concentrated to obtain a yellow oil, which was purified by flash column chromatography (PE:EA = 20:1) to obtain 5.4 g of white granular powder with a yield of 80%.

[0192] Example 1. Preparation of target compound BIOS-A1

[0193]

[0194] Step 1: Synthesis of intermediate tert-butyl 4-((2-bromo-6-chlorophenyl)sulfonamido)-1H-pyrazole-1-carboxylate (BIOS-A1-3)

[0195] 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 the reaction was completed, the reaction solution was washed with ammonium chloride solution 3 - 5 times. The organic phase was dried with anhydrous sodium sulfate and concentrated, and was purified by flash column chromatography to obtain a light gray solid BIOS-A1-3 (1.3 g, 3 mmol) with a yield of 60%.

[0196] ESI-MS: m / z = 436 [M+H] + 。

[0197] Step 2: Synthesis of Intermediate (S)-3-(1-phthalimidoethyl)-8-chloro-2-(1H-pyrazol-4-yl)-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A1-4):

[0198] Intermediate BIOS-A1-3 (1.3 g, 3 mmol), A-004 (720 mg, 3.6 mmol) and Pd(PPh 3 ) 4 (330 mg, 10 mol%) were placed in a dry sealed tube. The tube was purged with N 2 for 3 times, and then acetonitrile (30 mL) and DIPEA (1.2 g, 9 mmol) were added under positive pressure. The mixture was stirred at 90 °C overnight. After the raw materials disappeared, the solvent was evaporated to dryness to obtain a black oil, which was purified by flash column chromatography to give a pale yellow solid BIOS-A1-4 (454 mg, 1 mmol) with a yield of 33%.

[0199] ESI-MS: m / z = 455 [M+H] + .

[0200] 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)

[0201] Intermediate BIOS-A1-4 (454 mg, 1 mmol) and hydrazine hydrate (85%, 0.2 mL) were added to 10 mL of ethanol solution, and the mixture was stirred at 80 °C for 2 h. After the reaction was complete, it was cooled sufficiently, filtered by suction, and the filtrate was concentrated to obtain 200 mg of a white solid, which was directly used for the next reaction without purification.

[0202] ESI-MS: m / z = 325 [M+H] + .

[0203] 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)

[0204] 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 successively added to a single-necked flask, and the reaction was carried out at 90 °C for 8 h. The reaction mixture was concentrated under reduced pressure, and an appropriate amount of dichloromethane was added for extraction. Then, it was successively washed with saturated NaHCO 3Washed with aqueous solution, dilute hydrochloric acid (0.5 N) and saturated brine. The organic layer was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by preparative liquid chromatography to obtain the target product BIOS-A1 (46 mg), with a yield of 50%.

[0205] ESI-MS: m / z = 457 [M+H] + 。

[0206] 1 1H NMR (400 MHz, CDCl 3 ) δ 8.05 (s, 1H), 7.61 (s, 2H), 7.53 - 7.44 (m, 2H), 7.31 (dd, J = 6.9, 2.1 Hz, 1H), 6.53 (s, 1H), 5.56 (d, J = 7.2 Hz, 1H), 4.83 (t, J = 7.1 Hz, 1H), 2.63 (s, 3H), 1.54 (d, J = 7.0 Hz, 3H).

[0207] Example 2. Preparation of the target compound BIOS-A2

[0208]

[0209] Step 1: Synthesis of intermediate 2-bromo-6-chloro-N-(pyridin-3-yl)benzenesulfonamide (BIOS-A2-1):

[0210] Intermediate BIOS-A1-1 (1.4 g, 5 mmol) was dissolved in dichloromethane (20 mL), pyridine (2 mL) and 3-aminopyridine (450 mg, 5 mmol) were added, and the mixture was stirred at room temperature overnight until the raw materials reacted completely. The reaction was quenched with water, and the layers were separated. The organic phase was washed 3 times with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain BIOS-A2-1 (0.7 g), a white solid, with a yield of 50%.

[0211] ESI-MS: m / z = 347 [M+H] + 。

[0212] 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):

[0213] Intermediate BIOS-A2-1 (350 mg, 1 mmol), A-004 (240 mg, 1.2 mmol) and Pd(PPh 3 ) 4 (110 mg, 10 mol%) were placed in a dry sealed tube, N 2Replace three times. Add acetonitrile (20 mL) and DIPEA (400 mg, 3 mmol) under positive pressure, and stir at 90 °C overnight. After the raw materials disappeared, the solvent was evaporated to dryness to obtain a black oily substance, which was purified by flash column chromatography to obtain a pale yellow solid BIOS-A2-2 (200 mg), with a yield of 43%.

[0214] ESI-MS: m / z = 466 [M+H] + 。

[0215] 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):

[0216] Add intermediate BIOS-A2-2 (200 mg, 0.43 mmol) and hydrazine hydrate (85%, 0.3 mL) to an ethanol solution (10 mL). Stir the reaction at 80 °C for 2 h. After the reaction was complete, it was cooled sufficiently, filtered by suction, and the filtrate was concentrated to obtain a white solid BIOS-A2-3 (100 mg), which was directly used for the next reaction without purification.

[0217] ESI-MS: m / z = 336 [M+H] + 。

[0218] 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)

[0219] Add 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) to a single-necked flask in sequence, and react at 90 °C for 8 h until the raw materials reacted completely. Concentrate under reduced pressure, add an appropriate amount of dichloromethane for extraction, and wash successively with saturated NaHCO 3 aqueous solution, dilute hydrochloric acid (0.5 N) and saturated brine. The organic layer was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by preparative liquid phase to obtain the target product BIOS-A2 (40 mg), with a yield of 28%.

[0220] ESI-MS: m / z = 468 [M+H] + 。

[0221] 1 H NMR (400 MHz, DMSO-d 6)δ8.58 (dd, J = 4.4, 1.2 Hz, 1H), 8.38 (dd, J = 2.8, 0.8 Hz, 1H), 7.76 - 7.62 (m, 4H), 7.56 (d, J = 7.6 Hz, 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 = 6.8 Hz, 3H).

[0222] Example 3. Preparation of the target compound BIOS-A3

[0223]

[0224] Step 1: Synthesis of intermediate 2-bromo-6-chloro-N-(pyridin-4-yl)benzenesulfonamide (BIOS-A3-1)

[0225] Intermediate 2-bromo-6-chlorobenzenesulfonyl chloride (1.4 g, 5 mmol) was dissolved in dichloromethane (20 mL), pyridine (2 mL) and 3-aminopyridine (450 mg, 5 mmol) were added, and the mixture was stirred at room temperature overnight until the raw materials reacted completely. The reaction was quenched with water, and the layers were separated. The organic phase was washed 3 times with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by flash column chromatography to obtain BIOS-A3-1 (1.0 g), a white solid, with a yield of 60%.

[0226] ESI-MS: m / z = 347 [M + H] + .

[0227] 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)

[0228] Intermediate BIOS-A3-1 (700 mg, 2 mmol), alkynylamine derivative (480 mg, 2.4 mmol) and Pd(PPh 3 ) 4 (220 mg, 10 mol%) were placed in a dry sealed tube, and N 2 was replaced 3 times. Under positive pressure, acetonitrile (30 mL) and DIPEA (800 mg, 6 mmol) were added, and the mixture was stirred at 90 °C overnight. After the raw materials disappeared, the solvent was evaporated to dryness to obtain a black oil, which was purified by flash column chromatography to obtain 200 mg of a pale yellow solid, with a yield of 22%.

[0229] ESI-MS: m / z = 466 [M + H] + .

[0230] 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)

[0231] 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, it was cooled sufficiently, filtered by suction, and the filtrate was concentrated to obtain 100 mg of a white solid, which was directly used for the next reaction without purification.

[0232] ESI-MS: m / z = 336 [M+H] + 。

[0233] 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)

[0234] 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 successively added to a single-necked flask, and the reaction was carried out at 90 °C for 8 h. The reaction mixture was concentrated under reduced pressure, extracted with an appropriate amount of dichloromethane, and washed successively with saturated NaHCO 3 aqueous solution, dilute hydrochloric acid (0.5 N), and saturated brine. The organic layer was dried over anhydrous sodium sulfate, concentrated, and the residue was purified by high-performance liquid chromatography to obtain the target product BIOS-A3 (30 mg) with a yield of 21%.

[0235] ESI-MS: m / z = 468 [M+H] + 。

[0236] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 9.85 (d, J = 7.4 Hz, 1H), 8.58 (dd, J = 7.7, 2.0 Hz, 1H), 8.49 (dd, J = 7.6, 2.0 Hz, 1H), 8.40 - 8.23 (m, 2H), 7.38 (t, J = 7.7 Hz, 1H), 7.34 - 7.16 (m, 3H), 7.12 (dd, J = 7.9, 2.7 Hz, 1H), 6.09 (s, 1H), 4.61 - 4.54 (m, 1H), 2.54 (s, 3H), 1.54 (d, J = 6.7 Hz, 3H).

[0237] Example 4. Preparation of the target compound BIOS-A4

[0238]

[0239] Step 1: Preparation of 2-bromo-6-fluorobenzenesulfonyl chloride (BIOS-A4-2)

[0240] Under an ice bath, 2-bromo-6-fluoroaniline (3.85 g, 20.4 mmol) was added to 20 mL of concentrated hydrochloric acid, and 7 mL of sodium nitrite solution (2.8 g, 41 mmol) was slowly dropped in. In another reaction flask, copper sulfate pentahydrate (0.93 g, 3.7 mmol) and 32 mL of concentrated hydrochloric acid were added, and 20 mL of sodium bisulfite solution (7.05 g, 67.8 mmol) and the above solution were slowly dropped in under an ice bath. After the dropping was completed, the temperature was raised to room temperature and the reaction was carried out for 2 h until the raw materials reacted completely. The reaction solution was extracted with dichloromethane (80 mL * 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product BIOS-A4-2 (5.0 g). The product was directly used for the next step without purification.

[0241] LC-MS: 272.0 [M+H] + .

[0242] Step 2: Preparation of 2-bromo-6-fluoro-N-(pyridin-3-yl)benzenesulfonamide (BIOS-A4-3)

[0243] Under an ice bath, 3-aminopyridine (2.3 g, 24.1 mmol) was dissolved in 40 mL of pyridine, and BIOS-A4-2 (5.0 g) was dropped in. After the dropping was completed, the temperature was raised to room temperature and the reaction was carried out for 2 h until the raw materials reacted completely. After the reaction solution was concentrated under reduced pressure, 50 mL of dichloromethane was added, and it was washed twice with 1N HCl (50 mL * 2) and water (50 mL * 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue obtained was purified by silica gel column chromatography to obtain BIOS-A4-3 (3.6 g), with a yield of 59%.

[0244] LC-MS: 331.0 [M+H] + .

[0245] 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)

[0246] BIOS-A4-3 (3.6 g, 10.9 mmol), intermediate A-004 (3.3 g, 16.4 mmol) and Pd(PPh 3 )4 (5 mol%) was placed in a dry sealed tube, and N 2 was replaced three times. Under positive pressure, 50 mL of acetonitrile and DIPEA (32.7 mmol) were added, and the mixture was stirred at 90 °C overnight. After the raw materials disappeared, the solvent was evaporated to dryness to obtain a black oily substance, which was purified by flash column chromatography to obtain a pale yellow solid BIOS-A4-4 with a yield of 71%.

[0247] LC-MS: 450.0 [M+H] + 。

[0248] 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)

[0249] BIOS-A4-4 and hydrazine hydrate (85%, 1 mL, 15.5 mmol) were added to 50 mL of ethanol solution and cooled thoroughly. N 2 was replaced three times, and the mixture was stirred at 80 °C for 2 h. After the reaction was complete, it was cooled thoroughly, and a large amount of white flocculent precipitate was formed. It was filtered by suction, and the filtrate was concentrated to obtain a white solid. Ethanol solution was added thereto, cooled, and filtered by suction. The filtrate was further concentrated and ethyl acetate was added, cooled, and filtered by suction. The filtrate was evaporated to dryness to obtain a crude yellow oily product BIOS-A4-5, which was directly used for the next reaction without purification, with a yield of 85%.

[0250] LC-MS: 320.0 [M+H] + 。

[0251] 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)

[0252] The 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), DIPEA (387 mg, 3 mmol) were added, and N 2 was replaced three times, and the mixture was stirred at 90 °C overnight. The reaction solution was added to 10 mL of water and extracted with ethyl acetate (30 mL * 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 and purified by column chromatography to obtain the target product BIOS-A4 (280 mg), a yellow solid, with a yield of 62%.

[0253] LC-MS: 452.0 [M+H] + 。

[0254] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.58 (d, J = 4.8 Hz, 1H), 8.37 (d, J = 2.6 Hz, 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 = 7.0 Hz, 3H).

[0255] Example 5. Preparation of the target compound BIOS-A5

[0256]

[0257] Referring to Example 4, replace the compound 3-amino-pyridine in Step 2 of Example 4 with 5-fluoropyridine-3-amine, and keep other operations unchanged to obtain the compound BIOS-A5.

[0258] LC-MS: 470.1 [M + H] + .

[0259] Example 6. Preparation of the target compound BIOS-A6

[0260]

[0261]

[0262] Referring to Example 4, replace the compound BIOS-A4-1 in Step 1 of Example 4 with 2-bromo-6-chloroaniline, and replace the 3-amino-pyridine in Step 2 with 5-fluoropyridine-3-amine, and keep other operations unchanged to obtain the compound BIOS-A6 with a yield of 63%.

[0263] LC-MS: 486.0 [M + H] + .

[0264] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.62 (d, J = 2.6 Hz, 1H), 8.23 (d, J = 1.8 Hz, 1H), 7.81 - 7.60 (m, 4H), 7.49 (d, J = 7.7 Hz, 1H), 7.00 (s, 2H), 6.29 (s, 1H), 4.69 - 4.62 (m, 1H), 2.25 (s, 3H), 1.44 (d, J = 7.0 Hz, 3H).

[0265] Example 7. Preparation of the target compound BIOS-A7

[0266]

[0267] Step 1: Synthesis of Intermediate N-Benzyl-2-bromo-6-chlorobenzenesulfonamide (BIOS-A7-1)

[0268] 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 with ammonium chloride solution 3 - 5 times. The organic phase was dried over anhydrous sodium sulfate and concentrated, and then purified by flash column chromatography to obtain light gray solid BIOS-A7-1 (1.4 g) with a yield of 80%.

[0269] ESI-MS: m / z = 359 [M+H] + 。

[0270] 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)

[0271] BIOS-A7-1 (700 mg, 2 mmol), alkynylamine derivative (480 mg, 2.4 mmol) and Pd(PPh 3 ) 4 (220 mg, 10 mol%) were placed in a dry sealed tube, and N 2 was replaced 3 times. Under positive pressure, acetonitrile (30 mL) and DIPEA (800 mg, 6 mmol) were added, and the mixture was stirred at 90 °C overnight. After the raw materials disappeared, the solvent was evaporated to dryness to obtain a black oil, which was then purified by flash column chromatography to obtain light yellow solid BIOS-A7-2 (280 mg) with a yield of 30%.

[0272] ESI-MS: m / z = 479 [M+H] + 。

[0273] Step 3: Synthesis of (S)-3-(1-aminoethyl)-2-benzyl-8-chloro-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-A7-3)

[0274] Intermediate BIOS-A7-2 (280 mg, 0.6 mmol) and hydrazine hydrate (85%, 1.0 ml) were added to 15 mL of ethanol solution. The mixture was stirred at 80 °C for 2 h. After TLC showed that the reaction was complete, it was cooled sufficiently, filtered by suction, and the filtrate was concentrated to obtain 100 mg of white solid, which was directly used for the next reaction without purification.

[0275] ESI-MS: m / z = 349 [M+H]+ 。

[0276] 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)

[0277] Add 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) into a single-necked flask in sequence. React at 90 °C for 8 h until the raw materials react completely. Concentrate under reduced pressure, add an appropriate amount of dichloromethane for extraction, and wash successively with saturated NaHCO 3 aqueous solution, dilute hydrochloric acid (0.5 N) and saturated brine. The organic layer is dried over anhydrous sodium sulfate, concentrated, and the residue is purified by HPLC to obtain the target product BIOS-A7 (48 mg) with a yield of 33%.

[0278] ESI-MS: m / z = 481 [M+H] + 。

[0279] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.00 - 7.72 (m, 3H), 7.36 (m, 4H), 7.27 (m, 2H), 7.08 (d, J = 8.0 Hz, 1H), 6.70 (s, 1H), 5.74 (d, J = 10.4 Hz, 1H), 5.46 (d, = 18.2 Hz, 1H), 5.18 (d, J = 18.2 Hz, 1H), 5.11 - 5.05 (m, 1H), 2.22 (s, 3H), 0.70 (d, J = 6.5 Hz, 3H).

[0280] Example 8. Preparation of the target compound BIOS-A8

[0281]

[0282] Dissolve BIOS-A6 (100 mg, 0.2 mmol) in acetonitrile (2 mL), add PdCl 2 (CH 3 CN) 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), N 2Replace three times and reflux with stirring overnight. Add 10 mL of water to the reaction solution, extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated sodium chloride solution (20 mL × 1), dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify and separate by silica gel column chromatography to obtain BIOS-A8 (70 mg), a yellow solid, with a yield of 50%.

[0283] LC-MS: 691.2 [M+H] + 。

[0284] Example 9. Preparation of the target compound BIOS-B1

[0285]

[0286]

[0287] Step 1: Preparation of 2-chloro-6-methylbenzenesulfonyl chloride (BIOS-B1-2)

[0288] Under an ice bath, add 2-chloro-6-methylaniline (2.9 g, 20.4 mmol) to 20 mL of concentrated hydrochloric acid, and slowly dropwise add 7 mL of sodium nitrite solution (2.8 g, 41 mmol). In another reaction flask, add copper sulfate pentahydrate (0.93 g, 3.7 mmol) and 32 mL of concentrated hydrochloric acid, and slowly dropwise add 20 mL of sodium bisulfite solution (7.05 g, 67.8 mmol) and the above solution. After the addition is complete, raise the temperature to room temperature and react for 2 h. Extract the reaction solution with dichloromethane (80 mL × 3), dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure to obtain the crude product BIOS-B1-2 (4.0 g), and directly carry out the next reaction without purification of the product.

[0289] LC-MS: 225.0 [M+H] + 。

[0290] Step 2: Preparation of 2-chloro-6-methyl-N-phenylbenzenesulfonamide (BIOS-B1-3)

[0291] Under an ice bath, dissolve aniline (2.5 g, 24.1 mmol) in 40 mL of pyridine, and dropwise add BIOS-B1-2 (4.0 g). After the addition is complete, raise the temperature to room temperature and react for 2 h until the raw materials react completely. After concentrating the reaction solution under reduced pressure, add 50 mL of dichloromethane, wash twice with 1N HCl (50 mL × 2) and water (50 mL × 2), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify the residue obtained by silica gel column chromatography to obtain BIOS-B1-3 (3.6 g), with a yield of 72%.

[0292] LC-MS: 225.0 [M+H] + 。

[0293] 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)

[0294] Cool a solution of intermediate BIOS-B1-3 (1 g, 3.5 mmol) in anhydrous tetrahydrofuran (10 mL) to -78 °C, and then add a hexane solution of n-butyllithium (2.5 M, 4.3 mL). Stir the reaction mixture for 30 minutes. In another flask, cool a solution of tert-butyl (S)-(1-(methoxy(methyl)amino)-1-oxobutan-2-yl)carbamate (1.1 g, 4.55 mmol) in anhydrous tetrahydrofuran (10 mL). Slowly add a tetrahydrofuran solution of isopropylmagnesium chloride (2.5 mL, 5.3 mmol) at -78 °C. Stir the reaction mixture at the same temperature for 30 minutes, and then add it to the above reaction mixture. Stir at -78 °C for 1 hour. Quench the reaction mixture with water (50 mL), and then extract with ethyl acetate. Dry the extract over anhydrous sodium sulfate, and then concentrate it under reduced pressure. The product is directly subjected to the next reaction without purification.

[0295] LC-MS: 467.0 [M+H] + 。

[0296] Step 4: Preparation of (S)-3-(1-aminopropyl)-8-chloro-2-phenyl-2H-benzo[e][1,2]thiazine 1,1-dioxide (BIOS-B1-5)

[0297] Add the crude product of intermediate BIOS-B1-4 to a reaction flask, add 10 mL of 4 M hydrochloric acid dioxane solution, and stir at 100 °C for 4 h. After TLC shows that the reaction is complete, remove dioxane under reduced pressure, add a small amount of water, adjust the pH to alkaline with potassium carbonate, extract with ethyl acetate (50 mL * 2), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify and separate the product BIOS-B1-5 (700 mg) by silica gel column chromatography.

[0298] LC-MS: 349.0 [M+H] + 。

[0299] 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)

[0300] Dissolve the intermediate BIOS-B1-5 (700 mg, 2 mmol) in 7 mL of DMSO, add 2-amino-4-chloro-6-methylpyrimidine-5-carbonitrile (400 mg, 2.4 mmol), DIPEA (774 mg, 6 mmol), N 2 Replace three times, stir at 90 °C overnight until the raw materials react completely. Add 10 mL of water to the reaction solution, extract with ethyl acetate (30 mL * 3), combine the organic phases, wash with saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate and purify by silica gel column chromatography to obtain the target product BIOS-B1 (720 mg), a yellow solid, with a yield of 69%.

[0301] LC-MS: 481.0 [M+H] + 。

[0302] 1 H NMR (400 MHz, CDCl 3 ) δ 7.52 - 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 = 7.4 Hz, 3H).

[0303] Example 10. Preparation of the target compound BIOS-B2

[0304]

[0305] Referring to Example 9, replace the compound B-004-1 in step 3 of Example 9 with tert-butyl (S)-(4,4-difluoro-1-(methoxy(methyl)amino)-1-oxobutan-2-yl)carbamate, and keep other operations unchanged to obtain the compound BIOS-B2.

[0306] LC-MS: 517.1 [M+H] + 。

[0307] Example 11. Preparation of the target compound BIOS-B3

[0308]

[0309] Referring to Example 9, replace the compound B-004-1 in step 3 of the example with tert-butyl (S)-(1-cyclobutyl-2-(methoxy(methyl)amino)-2-oxoethyl)carbamate, and keep other operations unchanged to obtain the white powder target compound BIOS-B3 (25 mg).

[0310] LC-MS: 507.2 [M+H] + 。

[0311] Example 12. Preparation of the target compound BIOS-B4

[0312]

[0313] Referring to Example 9, replace the compound B-004-1 in Step 3 of the example with tert-butyl (S)-(1-(methoxy(methyl)amino)-3-methyl-1-oxobutan-2-yl)carbamate, and keep other operations unchanged to obtain the target compound BIOS-B4 as a white powder with a yield of 59%.

[0314] LC-MS: 495.0 [M+H] + 。

[0315] 1 H NMR (400 MHz, CDCl 3 ) δ 7.54 - 7.45 (m, 2H), 7.45 - 7.33 (m, 4H), 7.27 - 2.20 (m, 2H), 6.59 (s, 1H), 5.31 (d, J = 8.5 Hz, 1H), 4.97 (s, 2H), 4.61 (dd, J = 8.5, 4.6 Hz, 1H), 2.43 (s, 3H), 1.28 (s, 1H), 1.02 (d, J = 10.7, 6H).

[0316] Example 13. Preparation of the target compound BIOS-B5

[0317]

[0318] Referring to Example 9, replace the compound B-004-1 in Step 3 of the example with tert-butyl (S)-(1-cyclopropyl-2-(methoxy(methyl)amino)-2-oxoethyl)carbamate, and keep other operations unchanged to obtain the target compound BIOS-B5 as a white powder with a yield of 63%.

[0319] LC-MS: 493.0 [M+H] + 。

[0320] 1 H NMR (400 MHz, CDCl 3) δ 7.55 - 7.45 (m, 2H), 7.40 - 7.33 (m, 4H), 7.26 - 7.19 (m, 2H), 6.79 (s, 1H), 5.54 (d, J = 7.2 Hz, 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).

[0321] Example 14. Preparation of the target compound BIOS - B6

[0322]

[0323] Referring to Example 9, replace the compound B - 004 - 1 in step 3 of the example with tert - butyl (S) - 6 - (methoxy(methyl)carbamoyl)-5 - azaspiro[2.4]heptane - 5 - carboxylate, and keep other operations unchanged to obtain the compound BIOS - B6 with a yield of 25%.

[0324] ESI - MS: m / z = 519 [M + H] + 。

[0325] 1 1H NMR (400 MHz, DMSO - d 6 ) δ 7.70 - 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 = 6.0 Hz, 1H), 3.83 (d, J = 9.9 Hz, 1H), 3.67 (d, J = 9.8 Hz, 1H), 2.27 (s, 3H), 2.08 (dd, J = 12.4, 8.3 Hz, 1H), 1.81 (dd, J = 12.4, 2.4 Hz, 1H), 0.71 - 0.43 (m, 4H).

[0326] Example 15. Preparation of the target compound BIOS - B7

[0327]

[0328] Referring to Example 9, replace the compound B - 004 - 1 in step 3 of the example with tert - butyl (S) - 2 - (methoxy(methyl)carbamoyl)pyrrolidine - 1 - carboxylate, and keep other operations unchanged to obtain the white powder target compound BIOS - B7 with a yield of 82%.

[0329] LC - MS: 493.1 [M + H] + 。

[0330] 1 1H NMR (400 MHz, CDCl 3 ) δ 7.52 - 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 = 8.1 Hz, 1H), 3.92 - 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).

[0331] Example 16. Preparation of the target compound BIOS - B8

[0332]

[0333] Referring to Example 9, replace the compound 2 - amino - 4 - chloro - 6 - methylpyrimidine - 5 - carbonitrile in step 5 of Example 9 with 2,4 - diamino - 6 - chloropyrimidine - 5 - carbonitrile, and keep other operations unchanged. The target compound BIOS - B8 as a white powder is obtained with a yield of 30%.

[0334] LC - MS: 482.0 [M + H] + .

[0335] 1 1H NMR (400 MHz, DMSO - d 6 ) δ 7.82 - 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 = 7.2 Hz, 3H).

[0336] Example 17. Preparation of the target compound BIOS - B9

[0337]

[0338] Referring to Example 9, replace the compound 2 - chloro - 6 - methylaniline in step 1 of Example 9 with 2 - fluoro - 6 - methylaniline, and replace the compound aniline in step 2 with pyridin - 3 - ylmethanamine, and keep other operations unchanged. The target compound BIOS - B9 as a white powder is obtained.

[0339] ESI - MS: m / z = 480.1 [M + H] + .

[0340] Example 18. Preparation of the target compound BIOS-B10

[0341]

[0342] Referring to Example 9, replace the starting material 2-chloro-6-methylaniline in Step 1 of Example 9 with 2-fluoro-6-methylaniline (BIOS-B9-1) to generate the compound BIOS-B9-1. At the same time, replace aniline in Step 2 of Example 9 with (5-fluoropyridin-3-yl)methylamine, and keep other operations unchanged to obtain the compound BIOS-B10.

[0343] ESI-MS: m / z = 498.1 [M+H] + 。

[0344] Example 19. Preparation of the target compound BIOS-B11

[0345]

[0346] Dissolve BIOS-B1 (120 mg, 0.25 mmol) in acetonitrile (2 mL), add PdCl 2 (CH 3 CN) 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), replace N 2 three times, and reflux and stir overnight. Add 10 mL of water to the reaction solution, extract with ethyl acetate (30 mL * 3), combine the organic phases, wash with saturated sodium chloride solution (20 mL×1), dry over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and purify and separate by silica gel column chromatography to obtain BIOS-B11 (70 mg), a yellow solid, with a yield of 46%.

[0347] LC-MS: 603.2 [M+H] + 。

[0348] 1 H NMR (600 MHz, DMSO-d 6)δ 7.75 - 7.64 (m, 2H), 7.62 (dd, J = 7.3, 1.7 Hz, 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 (dd, J = 5.9, 3.7 Hz, 2H), 3.52 (dd, J = 5.8, 3.7 Hz, 2H), 3.48 (dd, J = 5.9, 3.7 Hz, 2H), 3.40 (dd, J = 5.8, 3.7 Hz, 2H), 3.22 (s, 3H), 2.25 (d, J = 1.9 Hz, 3H), 1.91 - 1.79 (m, 2H), 0.80 - 0.74 (m, 3H).

[0349] Example 20. Preparation of the target compound BIOS - B12

[0350]

[0351] Referring to Example 19, replace the compound BIOS - B11 - 1 in the steps of Example 19 with N,N - bis(2 - methoxyethyl)hex - 5 - ynamide, and keep other operations unchanged to obtain the light yellow powder compound BIOS - B12 with a yield of 67%.

[0352] LC - MS: 672.2 [M + H] + .

[0353] 1 1H NMR (400 MHz, CDCl 3 )δ 7.52 - 7.43 (m, 2H), 7.36 (dd, J = 5.2, 1.9 Hz, 3H), 7.31 (dd, J = 7.5, 1.5 Hz, 1H), 7.22 (dd, J = 6.7, 3.0 Hz, 2H), 6.55 (s, 1H), 5.21 (d, J = 7.9 Hz, 1H), 4.95 (s, 2H), 4.61 - 4.57 (m, 1H), 3.55 (t, J = 5.6 Hz, 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 = 7.4 Hz, 2H), 2.51 (t, J = 6.6 Hz, 2H), 2.39 (s, 3H), 1.95 - 1.90 (m, 2H), 1.76 - 1.71 (m, 2H), 0.97 (t, J = 7.4 Hz, 3H).

[0354] Example 21. Preparation of the target compound BIOS - B13

[0355]

[0356] Referring to Example 3, only replace 3-aminopyridine in Step 1 of Example 3 with aniline to prepare BIOS-B’ (i.e., Control Compound 2).

[0357] Referring to Example 19, replace the compound BIOS-B1 in the steps of Example 19 with BIOS-B’, and keep other operations unchanged to obtain the target compound BIOS-B13 as a pale yellow powder with a yield of 51%.

[0358] LC-MS: 589.1 [M+H] + 。

[0359] 1 H NMR (400 MHz, CDCl 3 ) δ 7.56 - 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 = 7.4 Hz, 1H), 4.98 (s, 2H), 4.76 - 4.71 (m, 1H), 4.43 (s, 2H), 3.77 (dd, J = 5.7, 3.7 Hz, 2H), 3.67 (dd, J = 5.8, 3.6 Hz, 2H), 3.63 (dd, J = 5.7, 3.8 Hz, 2H), 3.54 (dd, J = 5.8, 3.6 Hz, 2H), 3.39 - 3.34 (m, 3H), 2.40 (s, 3H), 1.49 (d, J = 6.9 Hz, 3H).

[0360] Example 22. Preparation of the target compound BIOS-B14

[0361]

[0362] Referring to Example 19, replace the compound BIOS-B11-1 in the steps of Example 19 with 4-ethynyl-1-methyl-1H-pyrazole, and keep other operations unchanged to obtain the target compound BIOS-B14 as a pale yellow powder with a yield of 73%.

[0363] LC-MS: 551.1 [M+H] + 。

[0364] 1 H NMR (400 MHz, CDCl 3)δ 7.67 - 7.49 (m, 4H), 7.42 - 7.32 (m, 4H), 7.26 - 7.23 (m, 2H), 6.59 (s, 1H), 5.23 (d, J = 7.8 Hz, 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 = 7.4 Hz, 3H).

[0365] Example 23. Preparation of the target compound BIOS - B15

[0366]

[0367] Referring to Example 19, replace the compound BIOS - B11 - 1 in the steps of Example 19 with 1 - morpholinocyclohex - 5 - yne - 1 - one, and keep other operations unchanged to obtain the compound BIOS - B15 with a yield of 69%.

[0368] LC - MS: 626.2 [M + H] + .

[0369] 1 H NMR (600 MHz, CDCl 3 )δ 7.52 - 7.46 (m, 2H), 7.41 - 7.36 (m, 3H), 7.32 (dd, J = 6.7, 2.4 Hz, 1H), 7.23 - 7.18 (m, 2H), 6.55 (s, 1H), 5.22 (d, J = 7.9 Hz, 1H), 4.97 (s, 2H), 4.60 - 4.56 (m, 1H), 3.63 (t, J = 4.9 Hz, 2H), 3.58 (dd, J = 5.7, 3.7 Hz, 2H), 3.53 (t, J = 4.8 Hz, 2H), 3.43 (dd, J = 5.8, 3.8 Hz, 2H), 2.60 (dt, J = 7.4, 2.7 Hz, 2H), 2.54 (dd, J = 7.1, 5.5 Hz, 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 = 7.4 Hz, 3H).

[0370] Example 24. Preparation of the target compound BIOS - B16

[0371]

[0372] Referring to Example 19, replace the compound BIOS-B11-1 in the steps of Example 19 with 1-(pyrrolidin-1-yl)hex-5-yn-1-one, and keep other operations unchanged to obtain compound BIOS-B16 with a yield of 67%.

[0373] LC-MS: 610.2[M+H] + 。

[0374] 1 H NMR(600MHz,CDCl 3 )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).

[0375] Example 25. Preparation of the target compound BIOS-B17

[0376]

[0377] Add BIOS-B1(150mg,0.31mmol), BIOS-B17-1(53mg,0.34mmol), potassium phosphate(131mg,0.62mmol), Pd(amphos) 2 Cl 2 (22mg,0.03mmol) and n-butanol:water = 2:1 (3mL in total) and 1mL of acetonitrile into the reaction flask. After replacing nitrogen, react at 80°C overnight. After the reaction is completed, remove the solvent under reduced pressure, add water and extract with ethyl acetate. Combine the organic phases, dry with anhydrous sodium sulfate and then concentrate. Purify and separate by silica gel column chromatography to obtain the target compound BIOS-B17 as an off-white powder with a yield of 83%.

[0378] LC-MS: 554.1[M+H] + 。

[0379] H NMR(400MHz,CDCl 3)δ8.16(dd, J = 5.2, 0.8Hz, 1H), 7.63(t, J = 7.7Hz, 1H), 7.50(dd, J

[0380] = 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), 3.94(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).

[0381] Synthesis of Control Compound 1. (S)-3-Amino-N-(1-(8-chloro-1,1-dihydroxy-2-phenyl-2H-benzo[e][1,2]thiazin-3-yl)ethyl)pyrazine-2-carboxamide

[0382]

[0383] Step 1: Synthesis of Intermediate 2-Bromo-6-chloro-N-phenylbenzenesulfonamide (1-1)

[0384] Intermediate 2-bromo-6-chlorobenzenesulfonyl chloride (1.4 g, 5 mmol) was dissolved in dichloromethane (20 mL), pyridine (2 mL) and aniline (450 mg, 5 mmol) were added, and the mixture was stirred at room temperature overnight. The reaction was quenched by adding water, and the layers were separated. The organic phase was washed 3 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), a white solid, with a yield of 80%.

[0385] ESI-MS: m / z = 346 [M+H] + .

[0386] 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)

[0387] Intermediate 1-1 (700 mg, 2 mmol), alkynylamine derivative (480 mg, 2.4 mmol) and Pd(PPh 3 ) 4 (220 mg, 10 mol%) were placed in a dry sealed tube, N 2It was changed 3 times. Under positive pressure, acetonitrile (30 mL) and DIPEA (800 mg, 6 mmol) were added, and the mixture was stirred at 90 °C overnight. After the raw materials disappeared, the solvent was evaporated to dryness to obtain a black oil, which was subjected to flash column chromatography to obtain a pale yellow solid 1-2 (280 mg) with a yield of 30%.

[0388] ESI-MS: m / z = 465 [M+H] + 。

[0389] Step 3: Synthesis of intermediate (S)-3-(1-aminoethyl)-8-chloro-2-phenyl-2H-benzo[e][1,2]thiazine 1,1-dioxide (1-3)

[0390] Intermediate 1-2 (280 mg, 0.6 mmol) and hydrazine hydrate (85%, 0.8 mL) were added to an ethanol solution (20 mL). The mixture was stirred at 80 °C for 2 h. After TLC showed that the reaction was complete, it was cooled sufficiently, filtered by suction, and the filtrate was concentrated to obtain 160 mg of a white solid, which was directly used for the next reaction without purification.

[0391] ESI-MS: m / z = 335 [M+H] + 。

[0392] 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)

[0393] 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 successively added to DMF. The mixture was stirred at room temperature for 2 h, and LC-MS showed that the reaction was complete. After the reaction solution was concentrated, it was directly purified by preparative liquid chromatography to obtain 68 mg of the target product as a white solid with a yield of 50%.

[0394] ESI-MS: m / z = 456 [M+H] + 。

[0395] 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.89 (d, J = 8.4 Hz, 1H), 8.23 (d, J = 2.3 Hz, 1H), 7.86 (d, J = 2.3 Hz, 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 = 7.0 Hz, 3H).

[0396] Example 26. ADP-Glo assay of the compounds of the present invention TM Kinase assay activity

[0397] ADP-Glo TM The ADP-Glo kinase assay was used to determine the inhibitory effect of the compound on PI3K kinase. ADP-Glo TM The assay consists of two steps. First, after the kinase reaction, an equal volume of ADP-Glo TM reagent was added to terminate the kinase reaction and deplete the remaining ATP. In the second step, a kinase detection reagent was added to convert ADP to ATP and allow the measurement of the newly synthesized ATP using the luciferase / luciferin reaction. The light produced was measured using a luminometer. By using the ATP-to-ADP conversion curve, the fluorescence could be correlated with the ADP concentration.

[0398] Experimental procedure: The test compound was diluted to a suitable series of concentrations (about 6 - 8 concentrations), and a source plate was prepared. The kinase reaction was carried out according to the kit, and the fluorescence signal was read. According to the formula inhibition rate = (maximum fluorescence - sample fluorescence) / (maximum fluorescence - minimum fluorescence) * 100, the inhibition percentage of each reaction relative to the DMSO-treated control was calculated, and then the 50% inhibitory concentration (IC 50 value) was calculated from the concentration-response curve.

[0399] Table 1: IC of PI3K kinase inhibition 50 (nM)

[0400]

[0401]

[0402] Note: “+++++” ≤ 5 nM; 5 nM < “++++” ≤ 10 nM; 10 nM < “+++” ≤ 100 nM; 100 < “++” ≤ 200 nM; “+” represents > 200 nM;

[0403] As can be seen from Table 1, the test compound showed significant inhibitory effects on PI3Kδ, and no or only low inhibitory effects on PI3Kα, PI3Kβ, and PI3Kγ.

[0404] Example 27. Inhibitory effect of some compounds on p-AKT in Joke-1 cells

[0405] TR-FRET combines time-resolved fluorescence detection technology and fluorescence energy resonance transfer detection (FRET) technology. In FRET experiments, biomolecules (such as proteins) are labeled with fluorescent donor-acceptor pairs. When biomolecules interact with each other, the distance between the donor and acceptor fluorescent groups is reduced. At this time, if the donor is excited, it will transfer the energy of its emitted light to the acceptor. The emitted light of the acceptor and donor has different wavelengths and can be distinguished by a microplate reader, thereby quantifying the interaction between biomolecules.

[0406] Add 8 μL of cell suspension to the detection plate. Add 2 μL of anti-IgM (250 ng / mL) and incubate at 30 °C for 60 minutes to stimulate JeKo-1 cells. Add compounds at different concentrations (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) to the cells and incubate at 37 °C for 10 minutes. Use the Phospho-AKT (Ser473) kit (Cisbio) to quantitatively determine p-AKT (a downstream product of PI3K) in the cells to judge the PI3Kδ inhibitory activity of the compounds, IC 50 The results are shown in Table 2.

[0407] Table 2: IC of PI3K kinase inhibition 50 (nM)

[0408] Compound Joke-1 cells BIOS-B5 +++ BIOS-A4 +++ BIOS-A6 +++ BIOS-B12 ++++ GSK2269557 (positive control) ++++

[0409]

Note

[0410] Example 28. Ovalbumin (OVA)-induced murine allergic asthma model

[0411] Female Balb / c mice at 6-8 weeks of age were intraperitoneally injected with ovalbumin sensitization solution (OVA 25 μg / mouse) on day 0 and day 14. To induce local inflammatory responses in the lungs, from day 21 to day 25, the mice were stimulated by nebulization with 1% OVA solution once a day for 30 min each time. After the animals were grouped, drug administration and challenge were carried out. One hour before each nebulization stimulation, the mice were anesthetized and fixed in the supine position on a 60° inclined plane, and dry powder nebulization drug administration was performed intratracheally, and lactose powder or compound BIOS-B12 (1.5 mg / kg) was given. On day 26, 24 hours after the last nebulization stimulation, the mice were subjected to a methacholine challenge test, and the airway reactivity of the mice was detected using an animal respiratory function detection system (EMMS-WBP), and the penh value was recorded. The results are as Figure 1As shown (MCH is acetylcholine). On the 27th day, after the mice were anesthetized, the neck skin was incised, the trachea in the neck was isolated, PBS was perfused into the lungs through the trachea and then aspirated back to obtain bronchoalveolar lavage fluid (BALF). The lavage was repeated 2 times, 0.6 mL each time. The BALF was centrifuged at 1500 r / min for 10 min at 4 °C. The cell pellet was resuspended with 1 mL PBS, and the total number of cells in the BALF of the mice was calculated using a cell counter. The results are as Figure 2 shown.

[0412] Ovalbumin can induce allergic asthma in mice. Under the stimulation of methacholine, the animals in the model control group showed obvious airway hyperresponsiveness. The detection results of Penh value showed that the administration of compound BIOS-B12 had a tendency to improve the airway hyperresponsiveness in mice. In addition, it was found that the administration of compound BIOS-B12 had an obvious inhibitory effect on the accumulation of cells in the bronchoalveolar lavage fluid after allergen stimulation.

[0413] Example 29. Pharmacokinetic study of the compound of the present invention by pulmonary administration

[0414] Male Sprague-Dawley rats were divided into a blank group, a group at 15 min, 30 min, 1 h, 2 h, and 4 h after administration, and were administered once. Blood samples were collected at the corresponding time points and plasma was separated; after blood collection, the animals were sacrificed, lung tissues were taken, lung tissue homogenates were prepared, the supernatant was centrifuged and taken, and the concentrations of B' (control compound 2) and BIOS-B12 in the lung tissues of Sprague-Dawley rats were determined by LC-MS / MS method. Pharmacokinetic parameters were calculated to investigate the metabolic characteristics of B' (control compound 2) and BIOS-B12 in the lung tissues of animals after administration in each group. The results are shown in Table 3.

[0415] Table 3: Pharmacokinetic data of the compound of the present invention

[0416]

[0417] It can be seen from Table 3 that compared with control compound 2, BIOS-B12 has a greater exposure and is more conducive to pulmonary inhalation administration.

[0418] Example 30. Pharmacokinetic study of the compound of the present invention by pulmonary administration (measurement of lung-blood ratio)

[0419] Male Sprague-Dawley (SD) rats were divided into a blank group and groups at 15 min, 30 min, 1 h, 2 h, and 4 h after drug administration. The rats were administered the drug once. Blood samples were collected at the corresponding time points, and plasma was separated. After blood collection, the animals were sacrificed, lung tissues were taken, lung tissue homogenates were prepared, the supernatant was taken after centrifugation, and the LC-MS / MS method was used to determine the concentrations of B’ (control compound 2), BIOS-B12, BIOS-B5, and BIOS-A3 in the plasma and lung tissues of SD rats. The lung-blood ratio was calculated by analyzing the pharmacokinetic parameters in the plasma and lung tissues, and the metabolic characteristics of B’ (control compound 2), BIOS-B12, BIOS-B5, and BIOS-A3 in animals after administration of each group were investigated.

[0420] The results are shown in Table 4.

[0421] Table 4: Lung-blood ratio data of the compounds of the present invention

[0422] Compound Lung-to-blood ratio (lung AUClast / plasma AUClast) BIOS-A3 277.23 BIOS-B5 227.48 BIOS-B12 233.78 Control compound 2 208.28

[0423] As can be seen from Table 4, compared with control compound 2, BIOS-B12, BIOS-B5, and BIOS-A3 have a greater lung-blood ratio and are more conducive to pulmonary inhalation administration.

[0424] Example 31. Inhibitory effect of the compound on the hERG ion channel

[0425] The hERG inhibitory effects of some compounds of the present invention were tested using a traditional model to judge safety, and the results are shown in Table 5.

[0426] Table 5: Blocking rate of the compound on the hERG ion channel

[0427] Compound 1 μM 10 μM BIOS-B12 1.6％ 17.46％

[0428] The results show that the compound BIOS-B12 of the present invention has no obvious inhibitory effect on the hERG channel.

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 Substitute, 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 Substitute, 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 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.

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.

Images