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CN116322697A - A kind of quinazoline compound and its pharmaceutical composition - Google Patents

A kind of quinazoline compound and its pharmaceutical compositionDownload PDF

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CN116322697A
CN116322697ACN202180064719.XACN202180064719ACN116322697ACN 116322697 ACN116322697 ACN 116322697ACN 202180064719 ACN202180064719 ACN 202180064719ACN 116322697 ACN116322697 ACN 116322697A
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alkylene
compound
substituted
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synthesis
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吴颢
路渊
陈小平
李波燕
周晓
傅水标
何将旗
王维
湛波
朱小惯
兰宏
王家炳
丁列明
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Betta Pharmaceuticals Co Ltd
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Abstract

A novel compound having cancer therapeutic activity. Also relates to a preparation method of the compounds and a pharmaceutical composition containing the compounds. (I)

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Translated fromChinese
一种喹唑啉化合物及其药物组合物A kind of quinazoline compound and its pharmaceutical composition技术领域technical field

本发明涉及一种新型化合物，其具有癌症治疗活性。本发明还涉及这些化合物的制备方法以及包含其的药物组合物。The present invention relates to a novel compound having cancer therapeutic activity. The invention also relates to processes for the preparation of these compounds and pharmaceutical compositions containing them.

技术背景technical background

临床数据显示，RAS是人类肿瘤中发生突变率最高的基因，所有肿瘤中，约20-30％有RAS突变，大约98％的胰腺癌，52％的结肠癌,43％的多发性骨髓瘤，及32％的肺腺癌中存在RAS基因突变。RAS最常见的突变方式是点突变，经常发生在12、13、61密码子，其中又以第12位密码子突变最常见。KRAS-G12C突变占KRAS突变的约10-20％，在非小细胞肺癌中占14％。Clinical data show that RAS is the gene with the highest mutation rate in human tumors. About 20-30% of all tumors have RAS mutations, about 98% of pancreatic cancer, 52% of colon cancer, 43% of multiple myeloma, And 32% of lung adenocarcinomas have RAS gene mutations. The most common mutation in RAS is point mutation, which often occurs atcodons 12, 13, and 61, and the mutation atcodon 12 is the most common. The KRAS-G12C mutation accounts for approximately 10-20% of KRAS mutations and 14% of non-small cell lung cancers.

40多年来，科学家一直在尝试开发靶向KRAS药物。AMGEN公司开发的AMG510是首个进入临床的KRAS G12C抑制剂。2020年12月，AMGEN公司向美国FDA递交AMG510的新药上市申请，用于治疗KRAS G12C突变的晚期转移性非小细胞肺癌。此前，FDA已经授予其突破性疗法认证资格。2021年5月，FDA批准全球首个KRAS靶向药AMG510上市。For more than 40 years, scientists have been trying to develop drugs that target KRAS. AMG510 developed by AMGEN is the first KRAS G12C inhibitor to enter the clinic. In December 2020, AMGEN submitted a new drug application for AMG510 to the US FDA for the treatment of advanced metastatic non-small cell lung cancer with KRAS G12C mutation. Previously, the FDA had granted it Breakthrough Therapy Designation. In May 2021, the FDA approved the world's first KRAS targeting drug AMG510 for marketing.

ARAXES公司专利WO2015054572A1记载一种具有特定轴手性片段的KRAS抑制剂，为外消旋体形式。该分子理论上具有R/S两种手性，但是专利未记载何种为优势活性构型。ARAXES patent WO2015054572A1 describes a KRAS inhibitor with a specific axial chiral fragment in the form of a racemate. The molecule theoretically has two chiralities of R/S, but the patent does not record which is the dominant active configuration.

Figure PCTCN2021124884-APPB-000001
Figure PCTCN2021124884-APPB-000001

劲方公司专利WO2020177629A1公开了特定手性分子的活性数据(表1)。根据说明书记载，具有相同轴手性片段的两对手性分子Z2-1/Z2-2和Z25-1/Z25-2，其活性较优的分子Z2-1和Z25-2的构型完全相反。The patent WO2020177629A1 of Jinfang Company discloses the activity data of specific chiral molecules (Table 1). According to the instructions, the two chiral molecules Z2-1/Z2-2 and Z25-1/Z25-2 with chiral fragments of the same axis have completely opposite configurations of the more active molecules Z2-1 and Z25-2.

Figure PCTCN2021124884-APPB-000002
Figure PCTCN2021124884-APPB-000002

表1Table 1

化合物编号Compound numberH358IC50(μM)H358IC50 (μM)A549IC50(μM)A549IC50 (μM)Z2Z20.0160.0163.2663.266Z2-2Z2-20.0230.0232.1652.165Z25Z250.0790.0792.7712.771Z25-2Z25-20.0440.0443.4243.424

发明内容Contents of the invention

本发明提供一种新型结构的手性KRAS抑制剂。预料不到的是，该手性分子活性优于其对应轴手性异构体，活性差异大于100倍。使用活性更优的手性分子，能够有效降低药物用量和避免药物不良反应，具有重要临床意义。The invention provides a chiral KRAS inhibitor with a novel structure. Unexpectedly, the activity of the chiral molecule is better than that of the corresponding axial chiral isomer, and the activity difference is more than 100 times. The use of chiral molecules with better activity can effectively reduce the dosage of drugs and avoid adverse drug reactions, which has important clinical significance.

本发明提供一种通式(I)所示的手性化合物、氘代物或药用盐，The present invention provides a chiral compound represented by general formula (I), a deuterated compound or a pharmaceutically acceptable salt,

Figure PCTCN2021124884-APPB-000003
Figure PCTCN2021124884-APPB-000003

其中X选自C3-14环烷基、3-14元杂环基、3-14元杂环基氨基；X任选地进一步被1-4个取代基取代，其中每个取代基独立地选自C1-6烷基、氨基、C1-6氨基烷基、氨基甲酰基、C1-6氨基甲酰基烷基、羧基、C1-6羧基烷基、氰基、C1-6氰基烷基、卤素、C1-6卤代烷基、羟基、C1-6羟烷基，所述C1-6烷基可任选地被一个或多个选自氨基、氨基甲酰基、羧基、氰基、卤素、羟基、氧代基所取代；Wherein X is selected from C3-14 cycloalkyl, 3-14 membered heterocyclyl, 3-14 membered heterocyclylamino; X is optionally further substituted by 1-4 substituents, wherein each substituent is independently selected from C1-6 alkyl, amino, C1-6 aminoalkyl, carbamoyl, C1-6 carbamoyl alkyl, carboxyl, C1-6 carboxyalkyl, cyano, C1-6 Cyanoalkyl, halogen, C1-6 haloalkyl, hydroxyl, C1-6 hydroxyalkyl, the C1-6 alkyl can optionally be selected from one or more of amino, carbamoyl, carboxyl , cyano, halogen, hydroxyl, oxo;

R1为丙烯酰基、取代的丙烯酰基或

Figure PCTCN2021124884-APPB-000004
R1 is acryloyl, substituted acryloyl or
Figure PCTCN2021124884-APPB-000004

R2选自H、C2-6烯基、C1-6烷氧基、C1-6卤代烷氧基、C1-6烷基、4-10元杂环基取代的C1-6烷基、C1-6烷基磺酰基、C1-6烷基亚磺酰基、C1-6烷硫基、C2-6炔基、C1-6烷基氨基、氨基、C1-6氨基烷基、氨基甲酰基、C1-6氨基甲酰基烷基、C1-6羧基烷基、氰基、C1-6氰基烷基、卤素、C1-6卤代烷基、取代或未取代的芳基、取代或未取代的5-10元杂芳基、取代或未取代的C3-7环烷基、取代或未取代的4-10元杂环基、羟基或氧代基，所述C1-6烷基可任选地被一个或多个选自氨基、氨基甲酰基、羧基、氰基、卤素、羟基、氧代基所取代；R2 is selected from H, C2-6 alkenyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 alkyl, 4-10 membered heterocyclic substituted C1-6 alkane C1-6 alkylsulfonyl, C 1-6 alkylsulfinyl, C1-6 alkylthio, C2-6 alkynyl,C 1-6alkylamino , amino, C1-6 Aminoalkyl, carbamoyl, C1-6 carbamoylalkyl, C1-6 carboxyalkyl, cyano, C1-6 cyanoalkyl, halogen, C1-6 haloalkyl, substituted or unsubstituted Substituted aryl, substituted or unsubstituted 5-10 membered heteroaryl, substituted or unsubstituted C3-7 cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclic group, hydroxyl or oxo group, The C1-6 alkyl group may optionally be substituted by one or more groups selected from amino, carbamoyl, carboxyl, cyano, halogen, hydroxyl, and oxo;

R3选自H、卤素、C1-6烷基、取代的C1-6烷基、C2-6烯基、取代的C2-6烯基、C3-6环烷基或取代的C3-6环烷基；R3 is selected from H, halogen, C1-6 alkyl, substituted C1-6 alkyl, C2-6 alkenyl, substituted C2-6 alkenyl, C3-6 cycloalkyl or substituted C3-6 cycloalkyl;

R4或R5独立地选自H、卤素、C1-6烷基、取代的C1-6烷基、C3-6环烷基、取代的C3-6环烷基；R4 or R5 are independently selected from H, halogen, C1-6 alkyl, substituted C1-6 alkyl, C3-6 cycloalkyl, substituted C3-6 cycloalkyl;

R6选自羟基、氧代基、C1-6烷氧基、C3-10环烷基氧基、氰基取代的环丙基C1-6亚烷基氧基，所述C1-6烷氧基任选地进一步被一个或多个选自卤素、羟基、C1-6烷氧基、C3-8环烷基的取代基所取代；R6 is selected from hydroxyl, oxo, C1-6 alkoxy, C3-10 cycloalkyloxy, cyano substituted cyclopropyl C1-6 alkyleneoxy, the C1- 6 alkoxy is optionally further substituted by one or more substituents selected from halogen, hydroxyl, C1-6 alkoxy, C3-8 cycloalkyl;

m或n独立地选自0、1、2、3、4；m or n are independently selected from 0, 1, 2, 3, 4;

Y选自3-14元杂环基，所述杂环基通过环上C原子与喹唑啉环直连相连，所述杂环基任选地进一步被1-4个选自R7或R8的取代基所取代；R7或R8独立地选自H、＝O、＝S、氰基、卤素、硝基、C1-6烷基、-C0-6亚烷基-ORa、-C0-6亚烷基-OC(O)N(Ra)2、-C0-6亚烷基-N(Ra)2、-C0-6亚烷基-NRaC(O)Ra、-C0-6亚烷基-NRaC(O)N(Ra)2、-C0-6亚烷基-NRaS(O)Ra、-C0-6亚烷基-NRaS(O)2Ra、-C0-6亚烷基-S(＝O)Ra、-C0-6亚烷基-S(＝O)2Ra、-C0-6亚烷基-SRa、-C0-6亚烷基-S(Ra)5、-C0-6亚烷基-C(＝O)Ra、-C0-6亚烷基-C(＝O)ORa、-C0-3亚烷基-C(＝O)N(Ra)2、C2-6烯基、C2-6炔基、丙烯酰基、-C0-6亚烷基-C3-14环烷基、-C0-6亚烷基-(3-14元杂环基)、-C0-6亚烷基-C6-14芳基或-C0-6亚烷基-(5-14元杂芳基)，所述C1-6烷基、C2-6烯基、C2-6炔基、-C0-6亚烷基-C3-14环烷基、-C0-6亚烷基-(3-14元杂环基)、-C0-6亚烷基-C6-14芳基或-C0-6亚烷基-(5-14元杂芳基)任选地还可被1个或多个Ra所取代；每个Ra各自独立地选自H、卤素、羟基、氨基、氧代基、硝基、氰基、羧基、C1-6烷基、C1-6羟基烷基、C1-6氨基烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C1-6杂烷基、C3-8环烷基、3-8元杂环基、C6-14芳基、5-14元杂芳基、C1-6酰基或

Figure PCTCN2021124884-APPB-000005
Y is selected from 3-14 membered heterocyclic groups, the heterocyclic group is directly connected to the quinazoline ring through the C atom on the ring, and the heterocyclic group is optionally further selected from 1-4 R7 or R8 is substituted by a substituent; R7 or R8 are independently selected from H, =O, =S, cyano, halogen, nitro, C1-6 alkyl, -C0-6 alkylene-ORa , -C0-6 alkylene-OC(O)N(Ra )2 , -C0-6 alkylene-N(Ra )2 , -C0-6 alkylene-NRa C( O)Ra , -C0-6 alkylene-NRa C(O)N(Ra )2 , -C0-6 alkylene-NRa S(O)Ra , -C0-6 Alkylene-NRa S(O)2 Ra , -C0-6 alkylene-S(=O)Ra , -C0-6 alkylene-S(=O)2 Ra ,- C0-6 alkylene-SRa , -C0-6 alkylene-S(Ra )5 , -C0-6 alkylene-C(=O)Ra , -C0-6 alkylene Alkyl-C(=O)ORa , -C0-3 alkylene-C(=O)N(Ra )2 , C2-6 alkenyl, C2-6 alkynyl, acryloyl, - C0-6 alkylene-C3-14 cycloalkyl, -C0-6 alkylene-(3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 aryl Or -C0-6 alkylene - (5-14 yuan heteroaryl), the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, -C0-6 alkylene Base-C3-14 cycloalkyl, -C0-6 alkylene-(3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 aryl or -C0-6 Alkylene-(5-14 membered heteroaryl) is optionally also substituted by 1 or more Ra ; each Ra is independently selected from H, halogen, hydroxyl, amino, oxo, Nitro, cyano, carboxyl, C1-6 alkyl, C1-6 hydroxyalkyl, C1-6 aminoalkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 Haloalkoxy, C1-6 heteroalkyl, C3-8 cycloalkyl, 3-8 membered heterocyclyl, C6-14 aryl, 5-14 membered heteroaryl, C1-6 acyl or
Figure PCTCN2021124884-APPB-000005

一些实施方式中，式(I)中的X选自4-10元杂环基，所述4-10元杂环基任选地进一步被1-4个选自C1-6烷基、氰基、C1-6氰基烷基、C1-6羟基烷基的取代基所取代；优选为

Figure PCTCN2021124884-APPB-000006
Figure PCTCN2021124884-APPB-000007
Figure PCTCN2021124884-APPB-000008
更优选为
Figure PCTCN2021124884-APPB-000009
In some embodiments, X in formula (I) is selected from 4-10 membered heterocyclic groups, and the 4-10 membered heterocyclic groups are optionally further replaced by 1-4 selected from C1-6 alkyl, cyano Substituents of radicals, C1-6 cyanoalkyl, C1-6 hydroxyalkyl; preferably
Figure PCTCN2021124884-APPB-000006
Figure PCTCN2021124884-APPB-000007
Figure PCTCN2021124884-APPB-000008
more preferably
Figure PCTCN2021124884-APPB-000009

一些实施方式中，式(I)中的R1选自

Figure PCTCN2021124884-APPB-000010
Figure PCTCN2021124884-APPB-000011
优选为
Figure PCTCN2021124884-APPB-000012
In some embodiments,R in formula (I) is selected from
Figure PCTCN2021124884-APPB-000010
Figure PCTCN2021124884-APPB-000011
preferably
Figure PCTCN2021124884-APPB-000012

一些实施方式中，式(I)中的R2选自H、卤素或C1-3烷基；优选为H。In some embodiments, R in formula (I)is selected from H, halogen or C1-3 alkyl; preferably H.

一些实施方式中，式(I)中的R3选自H、C1-6烷基、取代的C1-6烷基、C2-6烯基、取代的C2-6烯基、C3-6环烷基或取代的C3-6环烷基；优选C1-3烷基、C3-6环烷基或C2-4烯基；更优选为乙基、乙烯基或环丙基。In some embodiments,R in formula (I) is selected from H, C1-6 alkyl, substituted C1-6 alkyl, C2-6 alkenyl, substituted C2-6 alkenyl, C3-6 cycloalkyl or substituted C3-6 cycloalkyl; preferably C1-3 alkyl, C3-6 cycloalkyl or C2-4 alkenyl; more preferably ethyl, vinyl or cyclo Propyl.

一些实施方式中，式(I)中的R4或R5选自H、卤素或C1-6烷基；优选R4为H且R5为甲基。In some embodiments, R4 or R5 in formula (I) is selected from H, halogen or C1-6 alkyl; preferably R4 is H and R5 is methyl.

一些实施方式中，式(I)中的R6选自C1-6烷氧基、C3-8环烷基氧基、氰基取代的环丙基C1-6亚烷基氧基，所述C1-6烷氧基任选地进一步被一个或多个选自卤素、羟基、甲氧基、C3-8环烷基的取代基所取代；优选R6选自

Figure PCTCN2021124884-APPB-000013
Figure PCTCN2021124884-APPB-000014
In some embodiments,R in formula (I) is selected from C1-6 alkoxy, C3-8 cycloalkyloxy, cyclopropyl C1-6 alkyleneoxy substituted by cyano, The C1-6 alkoxy group is optionally further substituted by one or more substituents selected from halogen, hydroxyl, methoxy, and C3-8 cycloalkyl; preferablyR is selected from
Figure PCTCN2021124884-APPB-000013
Figure PCTCN2021124884-APPB-000014

一些实施方式中，式(I)中的Y选自

Figure PCTCN2021124884-APPB-000015
Figure PCTCN2021124884-APPB-000016
优选为
Figure PCTCN2021124884-APPB-000017
更优选为
Figure PCTCN2021124884-APPB-000018
In some embodiments, Y in formula (I) is selected from
Figure PCTCN2021124884-APPB-000015
Figure PCTCN2021124884-APPB-000016
preferably
Figure PCTCN2021124884-APPB-000017
more preferably
Figure PCTCN2021124884-APPB-000018

一些实施方式中，式(I)中的R7选自H、氰基、卤素、C1-6烷基、-C0-6亚烷基-ORa、-C0-6亚烷基-OC(O)N(Ra)2、-C0-6亚烷基-N(Ra)2、-C0-6亚烷基-NRaC(O)Ra、-C0-6亚烷基-NRaC(O)N(Ra)2、-C0-6亚烷基-NRaS(O)Ra、-C0-6亚烷基-NRaS(O)2Ra、-C0-6亚烷基-S(＝O)Ra、-C0-6亚烷基-S(＝O)2Ra、-C0-6亚烷基-SRa、-C0-6亚烷基-S(Ra)5、-C0-6亚烷基-C(＝O)Ra、-C0-6亚烷基-C(＝O)ORa、-C0-3亚烷基-C(＝O)N(Ra)2、丙烯酰基、-C0-6亚烷基-C3-14环烷基、-C0-6亚烷基-(3-14元杂环基)、-C0-6亚烷基-C6-14芳基或-C0-6亚烷基-(5-14元杂芳基)，所述C1-6烷基、-C0-6亚烷基-C3-14环烷基、-C0-6亚烷基-(3-14元杂环基)、-C0-6亚烷基-C6-14芳基或-C0-6亚烷基-(5-14元杂芳基)任选地还可被1个或多个Ra所取代；每个Ra各自独立地选自H、卤素、羟基、氨基、氧代基、氰基、羧基、C1-6烷基、C3-8环烷基、3-8元杂环基、C6-14芳基、5-14元杂芳基、C1-6酰基或

Figure PCTCN2021124884-APPB-000019
优选R7选自H、丙烯酰基、C1-6烷基、C1-6卤代烷基、C1-6酰基、-C0-3亚烷基-C1-6烷氧基、C1-6羟基烷基、C1-6氰基烷基、-C0-3亚烷基-S(＝O)2-C1-6烷基、C3-6酮基、
Figure PCTCN2021124884-APPB-000020
-C0-3亚烷基-C3-6环烷基、-C0-3亚烷基-C3-8杂环基、-C0-3亚烷基-苯基、-C0-3亚烷基-吡唑基，所述-C0-3亚烷基-C3-6环烷基、-C0-3亚烷基-C3-8杂环基、-C0-3亚烷基-苯基、-C0-3亚烷基-吡唑基任选地进一步被一个或多个选自C1-6酰基、羟基、C1-6烷基、C3-6环烷基、卤素、
Figure PCTCN2021124884-APPB-000021
或氰基地取代基所取代，所述C3-8杂环基含有一个选自O或N的杂原子；更优选R7选自H、C1-6烷基、乙酰基、
Figure PCTCN2021124884-APPB-000022
In some embodiments, R7 in formula (I) is selected from H, cyano, halogen, C1-6 alkyl, -C0-6 alkylene-ORa , -C0-6 alkylene- OC(O)N(Ra )2 , -C0-6 alkylene-N(Ra )2 , -C0-6 alkylene-NRa C(O)Ra , -C0-6 Alkylene-NRa C(O)N(Ra )2 , -C0-6 alkylene-NRa S(O)Ra , -C0-6 alkylene-NRa S(O)2 Ra , -C0-6 alkylene-S(=O)Ra , -C0-6 alkylene-S(=O)2 Ra , -C0-6 alkylene-SRa , -C0-6 alkylene-S(Ra )5 , -C0-6 alkylene-C(=O)Ra , -C0-6 alkylene-C(=O)ORa , -C0-3 alkylene-C(=O)N(Ra )2 , acryloyl, -C0-6 alkylene-C3-14 cycloalkyl, -C0-6 alkylene -(3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 aryl or -C0-6 alkylene-(5-14 membered heteroaryl), the C1 -6 alkyl, -C0-6 alkylene-C3-14 cycloalkyl, -C0-6 alkylene-(3-14 membered heterocyclyl), -C0-6 alkylene- C6-14 aryl or -C0-6 alkylene-(5-14 membered heteroaryl) optionally can also be substituted by 1 or more Ra ; each Ra is independently selected from H, halogen, hydroxyl, amino, oxo, cyano, carboxyl, C1-6 alkyl, C3-8 cycloalkyl, 3-8 membered heterocyclyl, C6-14 aryl, 5-14 Member heteroaryl, C1-6 acyl or
Figure PCTCN2021124884-APPB-000019
Preferably R is selected from H, acryloyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 acyl, -C0-3 alkylene-C1-6 alkoxy, C1-6 6 hydroxyalkyl, C1-6 cyanoalkyl, -C0-3 alkylene-S(=O)2 -C1-6 alkyl, C3-6 ketone,
Figure PCTCN2021124884-APPB-000020
-C0-3 alkylene-C3-6 cycloalkyl, -C0-3 alkylene-C3-8 heterocyclyl, -C0-3 alkylene-phenyl, -C0- 3 alkylene-pyrazolyl, the -C0-3 alkylene-C3-6 cycloalkyl, -C0-3 alkylene-C3-8 heterocyclyl, -C0-3 Alkylene-phenyl, -C0-3 alkylene-pyrazolyl is optionally further replaced by one or more selected from C1-6 acyl, hydroxyl, C1-6 alkyl, C3-6 ring Alkyl, halogen,
Figure PCTCN2021124884-APPB-000021
Or substituted by a cyano substituent, the C3-8 heterocyclic group contains a heteroatom selected from O or N; more preferably R7 is selected from H, C1-6 alkyl, acetyl,
Figure PCTCN2021124884-APPB-000022

一些实施方式中，式(I)中的R8选自H或C1-6烷基；优选为H。In some embodiments, R in formula (I ) is selected from H or C1-6 alkyl; preferably H.

一些实施方式中，通式(I)所示的手性化合物选自式(IA)-(ID)化合物，In some embodiments, the chiral compound represented by general formula (I) is selected from compounds of formula (IA)-(ID),

Figure PCTCN2021124884-APPB-000023
Figure PCTCN2021124884-APPB-000023

；优选为; preferably

Figure PCTCN2021124884-APPB-000024
Figure PCTCN2021124884-APPB-000024

其中，取代基如通式(I)所定义。Wherein, the substituents are as defined in general formula (I).

本发明还提供一种手性化合物、氘代物或药用盐，其选自：The present invention also provides a chiral compound, a deuterated compound or a pharmaceutically acceptable salt, which is selected from:

Figure PCTCN2021124884-APPB-000025
Figure PCTCN2021124884-APPB-000025

Figure PCTCN2021124884-APPB-000026
Figure PCTCN2021124884-APPB-000027
优选为
Figure PCTCN2021124884-APPB-000028
Figure PCTCN2021124884-APPB-000029
Figure PCTCN2021124884-APPB-000026
Figure PCTCN2021124884-APPB-000027
preferably
Figure PCTCN2021124884-APPB-000028
Figure PCTCN2021124884-APPB-000029

本发明提供一种药物组合物，所述药物组合物含有式(I)中的任一项所述的手性化合物、氘代物或药用盐和至少一种药学上可接受的辅料，药物组合物中所述手性化合物含量比其对应轴手性异构体含量更高。The present invention provides a pharmaceutical composition, which contains the chiral compound, deuterium compound or pharmaceutically acceptable salt described in any one of formula (I) and at least one pharmaceutically acceptable excipient, the pharmaceutical composition The content of the chiral compound in the compound is higher than that of the corresponding axial chiral isomer.

本发明还提供一种药物组合物，药物组合物中所述通式(I)所示的手性化合物、氘代物、药用盐在活性成分中的含量高于51％、61％、71％、81％、91％、99％或更高。The present invention also provides a pharmaceutical composition, in which the content of the chiral compound, deuterium compound, and pharmaceutically acceptable salt represented by the general formula (I) in the active ingredient is higher than 51%, 61%, or 71%. , 81%, 91%, 99% or higher.

本发明还提供一种药物组合物，药物组合物中治疗有效量的至少一种式(I)所示的化合物、氘代物、药用盐和药学上可接受的辅料的质量百分比为0.0001:1-10。The present invention also provides a pharmaceutical composition, the therapeutically effective amount of at least one compound represented by formula (I), deuterium, pharmaceutically acceptable salt and pharmaceutically acceptable adjuvant in the pharmaceutical composition is 0.0001:1 by mass -10.

本发明还提供通式(I)中手性化合物、氘代物、药用盐或含有结构式(I)所示化合物的药物组合物在制备药物中的应用；优选所述药物为抗癌药物。The present invention also provides the use of chiral compounds, deuterated compounds, pharmaceutically acceptable salts in general formula (I), or pharmaceutical compositions containing compounds represented by structural formula (I) in the preparation of medicines; preferably, the medicines are anticancer drugs.

作为优选，所述应用为制备用于治疗由KRAS G12C介导的疾病的药物的应用。作为优选，所述疾病是癌症。Preferably, the application is the application of preparing medicines for treating diseases mediated by KRAS G12C. Preferably, said disease is cancer.

作为优选，所述癌症选自乳腺癌、多发性骨髓瘤、膀胱癌、子宫内膜癌、胃癌、宫颈癌、横纹肌肉瘤、非小细胞肺癌、小细胞肺癌、多形性肺癌、卵巢癌、食管癌、黑色素瘤、结肠直肠癌、肝细胞瘤、头颈部肿瘤、肝胆管细胞癌、骨髓增生异常综合征、恶性胶质瘤、前列腺癌、甲状腺癌、徐旺氏细胞瘤、肺鳞状细胞癌、苔藓样角化病、滑膜肉瘤、皮肤癌、胰腺癌、睾丸癌或脂肪肉瘤。Preferably, the cancer is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic lung cancer, ovarian cancer, esophagus Carcinoma, melanoma, colorectal cancer, hepatoma, head and neck tumors, hepatocholangiocarcinoma, myelodysplastic syndrome, glioblastoma, prostate cancer, thyroid cancer, Schwann cell tumor, lung squamous cell Carcinoma, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer, or liposarcoma.

本发明还提供一种治疗和/或预防癌症的方法，包括向治疗对象施用治疗有效量的通式(I)中手性化合物、氘代物、药用盐或含有结构式(I)所示化合物的药物组合物；优选所述癌症是由KRAS G12C，HRAS G12C或NRAS G12C突变介导的。The present invention also provides a method for treating and/or preventing cancer, comprising administering a therapeutically effective amount of a chiral compound, a deuterated substance, a pharmaceutically acceptable salt in the general formula (I) or a drug containing a compound represented by the structural formula (I) to the treatment subject. A pharmaceutical composition; preferably said cancer is mediated by KRAS G12C, HRAS G12C or NRAS G12C mutations.

作为优选，在上述方法中，所述的癌症选自乳腺癌、多发性骨髓瘤、膀胱癌、子宫内膜癌、胃癌、宫颈癌、横纹肌肉瘤、非小细胞肺癌、小细胞肺癌、多形性肺癌、卵巢癌、食管癌、黑色素瘤、结肠直肠癌、肝细胞瘤、头颈部肿瘤、肝胆管细胞癌、骨髓增生异常 综合征、恶性胶质瘤、前列腺癌、甲状腺癌、徐旺氏细胞瘤、肺鳞状细胞癌、苔藓样角化病、滑膜肉瘤、皮肤癌、胰腺癌、睾丸癌或脂肪肉瘤。Preferably, in the above method, the cancer is selected from breast cancer, multiple myeloma, bladder cancer, endometrial cancer, gastric cancer, cervical cancer, rhabdomyosarcoma, non-small cell lung cancer, small cell lung cancer, pleomorphic Lung cancer, ovarian cancer, esophageal cancer, melanoma, colorectal cancer, hepatocellular carcinoma, head and neck cancer, hepatocholangiocarcinoma, myelodysplastic syndrome, malignant glioma, prostate cancer, thyroid cancer, Schwann cell tumor, squamous cell carcinoma of the lung, lichenoid keratosis, synovial sarcoma, skin cancer, pancreatic cancer, testicular cancer, or liposarcoma.

除非另有说明，所述结构通式中使用的一般化学术语具有通常的含义。Unless otherwise stated, general chemical terms used in the structural formulae have their usual meanings.

例如，除非另有说明，本发明所用的术语“卤素”是指氟、氯、溴或碘。For example, the term "halogen" as used herein refers to fluorine, chlorine, bromine or iodine, unless otherwise specified.

在本发明中，除非另有说明，“烷基”包括直链或支链的一价饱和烃基。例如，烷基包括甲基、乙基、丙基、异丙基、正丁基、异丁基、仲丁基、叔丁基、正戊基、3-(2-甲基)丁基、2-戊基、2-甲基丁基、新戊基、正己基、2-己基、2-甲基戊基等。类似的，“C1-8烷基”中的“1-8”是指包含有1、2、3、4、5、6、7或8个碳原子的直链或支链形式排列的基团。In the present invention, unless otherwise specified, "alkyl" includes linear or branched monovalent saturated hydrocarbon groups. For example, alkyl includes methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 3-(2-methyl)butyl, 2 -pentyl, 2-methylbutyl, neopentyl, n-hexyl, 2-hexyl, 2-methylpentyl and the like. Similarly, "1-8 " in "C1-8 alkyl" refers to a group containing 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms arranged in a straight or branched chain group.

“C1-3亚烷基”是指包括直链或支链的二价饱和烃基。例如亚甲基、1,2-亚乙基、1,3-亚丙基或1,2-亚异丙基。类似的，“C1-6亚烷基”中的“1-6”是指包含有1、2、3、4、5或6个碳原子的直链或支链形式排列的基团。The "C1-3 alkylene group" refers to a divalent saturated hydrocarbon group including straight chain or branched chain. For example methylene, 1,2-ethylene, 1,3-propylene or 1,2-isopropylene. Similarly, "1-6 " in "C1-6 alkylene" refers to a group comprising 1, 2, 3, 4, 5 or 6 carbon atoms arranged in a linear or branched chain.

在本发明中，“一”、“一个”、“该”、“至少一个”和“一个或多个”可互换使用。因此，例如，包含“一种”药学上可接受的赋形剂的组合物可以被解释为表示该组合物包括“一种或多种”药学上可接受的赋形剂。In the present invention, "a", "an", "the", "at least one" and "one or more" are used interchangeably. Thus, for example, a composition comprising "a" pharmaceutically acceptable excipient may be interpreted to mean that the composition includes "one or more" pharmaceutically acceptable excipients.

术语“芳基”，在本发明中，除非另有说明，是指未取代或取代的包括碳环的原子的单环或稠环芳香基团，具有完全共轭的π电子体系。优选芳基为6-14元的单环或多环的芳香环基团。优选为苯基、萘基。最优选为苯基。所述芳基环可以稠合于杂芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为芳基环。The term "aryl", in the present invention, unless otherwise stated, refers to an unsubstituted or substituted monocyclic or condensed ring aromatic group comprising atoms of a carbocyclic ring, having a fully conjugated π-electron system. Preferably, the aryl group is a 6-14 membered monocyclic or polycyclic aromatic ring group. Preferred are phenyl and naphthyl. Most preferred is phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, where the ring bonded to the parent structure is the aryl ring.

术语“杂环基”，在本发明中，除非另有说明，是指由碳原子和1-3个选自N、O或S的杂原子组成的未取代或取代的3-14元稳定环系统，其为饱和或部分不饱和单环或多环环状烃取代基，如螺环或桥环。杂环基中氮或硫杂原子可以选择性地被氧化，并且氮杂原子可以选择性地被季铵化。该杂环基可以被连接到任何的杂原子或碳原子上以形成稳定的结构。这些杂环基的实例包括但不限于氮杂环丁烷基、吡咯烷基、哌啶基、哌嗪基、氧代哌嗪基、氧代哌啶基、四氢呋喃基、二氧戊环基、四氢咪唑基、四氢噻唑基、四氢恶唑基、四氢吡喃基、吗啉基、硫代吗啉基、硫代吗啉基亚砜、硫代吗啉基砜基和四氢恶二唑基。所述杂环基可以稠合于芳基、杂芳基或环烷基环上，其中与母体结构连接在一起的环为杂环基。The term "heterocyclyl", in the present invention, unless otherwise specified, refers to an unsubstituted or substituted 3-14 membered stable ring consisting of carbon atoms and 1-3 heteroatoms selected from N, O or S systems, which are saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituents, such as spiro or bridged rings. The nitrogen or sulfur heteroatoms in the heterocyclyl group can be optionally oxidized, and the nitrogen heteroatoms can be optionally quaternized. The heterocyclyl group can be attached to any heteroatom or carbon atom to form a stable structure. Examples of such heterocyclic groups include, but are not limited to, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxopiperazinyl, oxopiperidinyl, tetrahydrofuranyl, dioxolanyl, Tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydrooxazolyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone and tetrahydro Oxadiazolyl. The heterocyclyl may be fused to an aryl, heteroaryl or cycloalkyl ring, where the ring bonded to the parent structure is the heterocyclyl.

术语“螺环基”，在本发明中，除非另有说明，是指两个单环共用一个碳原子的多环化合物，这些螺环基的实例包括但不限于

Figure PCTCN2021124884-APPB-000030
Figure PCTCN2021124884-APPB-000031
The term "spirocyclic group", in the present invention, unless otherwise stated, refers to a polycyclic compound in which two monocyclic rings share one carbon atom. Examples of these spirocyclic groups include but are not limited to
Figure PCTCN2021124884-APPB-000030
Figure PCTCN2021124884-APPB-000031

术语“桥环基”，在本发明中，除非另有说明，是指两个单环共用两个或两个以上碳原子的多环化合物，这些桥环基的实例包括但不限于

Figure PCTCN2021124884-APPB-000032
The term "bridged ring group", in the present invention, unless otherwise specified, refers to a polycyclic compound in which two monocyclic rings share two or more carbon atoms. Examples of these bridged ring groups include but are not limited to
Figure PCTCN2021124884-APPB-000032

术语“杂芳基”，在本发明中，除非另有说明，是指未取代或取代的稳定的5元或6元单环芳族环系统或未取代或取代的9-14元苯并稠合杂芳族环系统或多环杂芳族环系统，其由碳原子和1-4个选自N、O或S的杂原子组成，并且其中所述氮或硫杂原子可以选择性地被氧化，所述氮杂原子可以选择性地被季铵化。杂芳基可以连接在任何杂原子或碳原子上以形成稳定的结构。杂芳基的实例包括但不限于噻吩基、呋喃基、咪唑基、异恶唑基、恶唑基、吡唑基、吡咯基、噻唑基、噻二唑基、三唑基、吡啶基、哒嗪基、吲哚基、氮杂吲哚基、吲唑基、苯并咪唑基、苯并呋喃基、苯并噻吩基、苯并异恶唑基、苯并噻唑基、苯并噻唑基、苯并噻二唑基、苯并三唑基腺嘌呤、喹啉基或异喹啉基。所述杂芳基可以稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环。The term "heteroaryl", in the present invention, unless otherwise stated, refers to an unsubstituted or substituted stable 5- or 6-membered monocyclic aromatic ring system or an unsubstituted or substituted 9-14-membered benzofused A heteroaromatic ring system or a polycyclic heteroaromatic ring system consisting of carbon atoms and 1-4 heteroatoms selected from N, O or S, and wherein the nitrogen or sulfur heteroatoms can be optionally replaced by Oxygenated, the nitrogen heteroatoms can optionally be quaternized. Heteroaryl groups can be attached to any heteroatom or carbon atom to form a stable structure. Examples of heteroaryl groups include, but are not limited to, thienyl, furyl, imidazolyl, isoxazolyl, oxazolyl, pyrazolyl, pyrrolyl, thiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridyl, Azinyl, indolyl, azaindolyl, indazolyl, benzimidazolyl, benzofuryl, benzothienyl, benzisoxazolyl, benzothiazolyl, benzothiazolyl, benzene Thiadiazolyl, benzotriazolyladenine, quinolinyl or isoquinolinyl. The heteroaryl group may be fused to an aryl, heterocyclyl or cycloalkyl ring where the ring attached to the parent structure is a heteroaryl ring.

术语“环烷基”是指具有3-14个碳原子的环状饱和或部分不饱和单环或多环环状烃取代基，例如，环丙基、环丁基、环戊基或环己基。所述环烷基可以稠合于芳基、杂环基或杂芳基环上，其中与母体结构连接在一起的环为环烷基。The term "cycloalkyl" refers to a cyclic saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent having 3 to 14 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl . The cycloalkyl group may be fused to an aryl, heterocyclyl, or heteroaryl ring where the ring bonded to the parent structure is a cycloalkyl group.

术语“取代的”是指基团中的一个或多个氢原子分别被相同的或者不同的取代基所取代。典型的取代基包括但不限于H、＝O、＝S、氰基、卤素、硝基、C1-6烷基、-C0-6亚烷基-ORa、-C0-6亚烷基-OC(O)N(Ra)2、-C0-6亚烷基-N(Ra)2、-C0-6亚烷基-NRaC(O)Ra、-C0-6亚烷基-NRaC(O)N(Ra)2、-C0-6亚烷基-NRaS(O)Ra、-C0-6亚烷基-NRaS(O)2Ra、-C0-6亚烷基-S(＝O)Ra、-C0-6亚烷基-S(＝O)2Ra、-C0-6亚烷基-SRa、-C0-6亚烷基-S(Ra)5、-C0-6亚烷基-C(＝O)Ra、-C0-6亚烷基-C(＝O)ORa、-C0-3亚烷基-C(＝O)N(Ra)2、C2-6烯基、C2-6炔基、丙烯酰基、-C0-6亚烷基-C3-14环烷基、-C0-6亚烷基-(3-14元杂环基)、-C0-6亚烷基-C6-14芳基或-C0-6亚烷基-(5-14元杂芳基)，所述C1-6烷基、C2-6烯基、C2-6炔基、-C0-6亚烷基-C3-14环烷基、-C0-6亚烷基-(3-14元杂环基)、-C0-6亚烷基-C6-14芳基或-C0-6亚烷基-(5-14元杂芳基)任选地还可被1个或多个Ra所取代；每个Ra各自独立地选自H、卤素、羟基、氨基、氧代基、硝基、氰基、羧基、C1-6烷基、C1-6羟基烷基、C1-6氨基烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、 C1-6杂烷基、C3-8环烷基、3-8元杂环基、C6-14芳基、5-14元杂芳基、C1-6酰基或

Figure PCTCN2021124884-APPB-000033
在一些实施例中，取代基独立地选自包含-F、-Cl、-Br、-I、-OH、三氟甲氧基、乙氧基、丙氧基、异丙氧基、正丁氧基、异丁氧基、叔丁氧基、-SCH3、-SC2H5、甲醛基、-C(OCH3)、氰基、硝基、-CF3、-OCF3、氨基、二甲基氨基、甲硫基、磺酰基和乙酰基的基团。The term "substituted" means that one or more hydrogen atoms in the group are respectively replaced by the same or different substituents. Typical substituents include, but are not limited to, H, =O, =S, cyano, halogen, nitro, C1-6 alkyl, -C0-6 alkylene-ORa , -C0-6 alkylene -OC(O)N(Ra )2 , -C0-6 alkylene-N(Ra )2 , -C0-6 alkylene-NRa C(O)Ra , -C0 -6 alkylene-NRa C(O)N(Ra )2 , -C0-6 alkylene-NRa S(O)Ra , -C0-6 alkylene-NRa S( O)2 Ra , -C0-6 alkylene-S(=O)Ra , -C0-6 alkylene-S(=O)2 Ra , -C0-6 alkylene- SRa , -C0-6 alkylene-S(Ra )5 , -C0-6 alkylene-C(=O)Ra , -C0-6 alkylene-C(=O) ORa , -C0-3 alkylene-C(=O)N(Ra )2 , C2-6 alkenyl, C2-6 alkynyl, acryloyl, -C0-6 alkylene- C3-14 cycloalkyl, -C0-6 alkylene-(3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 aryl or -C0-6 alkylene Base-(5-14 membered heteroaryl), the C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, -C0-6 alkylene-C3-14 cycloalkane Base, -C0-6 alkylene-(3-14 membered heterocyclic group), -C0-6 alkylene-C6-14 aryl or -C0-6 alkylene-(5-14 (Heteroaryl) can optionally be substituted by one or more Ra ; each Ra is independently selected from the group consisting of H, halogen, hydroxyl, amino, oxo, nitro, cyano, carboxyl, C1-6 alkyl, C1-6 hydroxyalkyl, C1-6 aminoalkyl, C1-6 haloalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 Heteroalkyl, C3-8 cycloalkyl, 3-8 membered heterocyclic group, C6-14 aryl, 5-14 membered heteroaryl, C1-6 acyl or
Figure PCTCN2021124884-APPB-000033
In some embodiments, the substituents are independently selected from the group consisting of -F, -Cl, -Br, -I, -OH, trifluoromethoxy, ethoxy, propoxy, isopropoxy, n-butoxy Butyl, isobutoxy, tert-butoxy, -SCH3 , -SC2 H5 , formaldehyde, -C(OCH3 ), cyano, nitro, -CF3 , -OCF3 , amino, dimethyl Amino, methylthio, sulfonyl and acetyl groups.

取代烷基的实例包括但不限于2,3-二羟基丙基、2-氨基乙基、2-羟乙基、五氯乙基、三氟甲基、甲氧基甲基、五氟乙基、苯基甲基、二恶茂基甲基和哌嗪基甲基。Examples of substituted alkyl groups include, but are not limited to, 2,3-dihydroxypropyl, 2-aminoethyl, 2-hydroxyethyl, pentachloroethyl, trifluoromethyl, methoxymethyl, pentafluoroethyl , phenylmethyl, dioxolylmethyl and piperazinylmethyl.

取代烷氧基的实例包括但不限于2-羟基乙氧基、2-氟乙氧基、2,2-二氟乙氧基、2-甲氧基乙氧基、2-氨基乙氧基、2,3-二羟基丙氧基、环丙基甲氧基、氨基甲氧基、三氟甲氧基、2-二乙基氨基乙氧基、2-乙氧基羰基乙氧基、3-羟基丙氧基。Examples of substituted alkoxy groups include, but are not limited to, 2-hydroxyethoxy, 2-fluoroethoxy, 2,2-difluoroethoxy, 2-methoxyethoxy, 2-aminoethoxy, 2,3-dihydroxypropoxy, cyclopropylmethoxy, aminomethoxy, trifluoromethoxy, 2-diethylaminoethoxy, 2-ethoxycarbonylethoxy, 3- Hydroxypropoxyl.

术语“羟基烷基”是指一个或多个羟基取代的烷基，例如-(亚烷基)-OH。The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups, eg -(alkylene)-OH.

术语“卤代烷基”是指一个或多个卤素取代的烷基，例如-(亚烷基)-卤素。The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, eg -(alkylene)-halogen.

术语“烷氧基”是指-O-(烷基)。The term "alkoxy" refers to -O-(alkyl).

术语“药学上可接受的盐”是指从药学上可接受的无毒的碱或酸制备的盐。The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid.

由于式(I)所示化合物将作为药物应用，较优地，使用一定纯度，例如，至少为60％纯度，比较合适的纯度为至少75％，特别合适地纯度为至少98％(％是重量比)。Since the compound shown in formula (I) will be used as a medicine, preferably, a certain purity is used, for example, at least 60% purity, more suitable purity is at least 75%, particularly suitable purity is at least 98% (% is weight) Compare).

本发明化合物的药物前体包含在本发明的保护范围内。通常，所述药物前体是指很容易在体内转化成所需化合物的功能性衍生物。例如，本申请化合物的任何药学上可接受的盐、酯、酯的盐或其它衍生物，其在向受体施用后能够直接或间接地提供本申请的化合物或其具有药学活性的代谢物或残基。特别优选的衍生物或前药是在施用于患者时可以提高本申请化合物生物利用度的那些化合物(例如，可以使口服的化合物更易于被吸收到血液中)，或者促进母体化合物向生物器官或作用位点(例如脑部或淋巴系统)递送的那些化合物。因此，本发明提供的治疗方法中的术语“给药”是指施用能治疗不同疾病的本发明公开的化合物，或虽未明确公开但对受试者给药后能够在体内转化为本发明公开的化合物的化合物。The prodrugs of the compounds of the present invention are included in the protection scope of the present invention. In general, the prodrugs refer to functional derivatives that are readily converted in vivo into the desired compound. For example, any pharmaceutically acceptable salt, ester, ester salt or other derivative of the compound of the present application, which can directly or indirectly provide the compound of the present application or its pharmaceutically active metabolite or Residues. Particularly preferred derivatives or prodrugs are those compounds which, when administered to a patient, increase the bioavailability of the compounds of the present application (e.g., allow an orally administered compound to be more readily absorbed into the blood), or promote the delivery of the parent compound to a biological organ or Those compounds delivered at a site of action such as the brain or lymphatic system. Therefore, the term "administering" in the treatment methods provided by the present invention refers to the administration of the compounds disclosed in the present invention that can treat different diseases, or the compounds disclosed in the present invention, although not explicitly disclosed, can be converted into compounds disclosed in the present invention after administration to the subject. compounds of compounds.

显然的，一个分子中任何取代基或特定位置的变量的定义是独立于分子中其他位置的。很容易理解，本领域技术人员可以通过现有技术手段及本发明中所述的方法来选择本发明中的化合物的取代基或取代形式，以获得化学上稳定且易于合成的化合物。Clearly, any substituent or variable at a particular position in a molecule is defined independently of other positions in the molecule. It is easy to understand that those skilled in the art can select the substituents or substitution forms of the compounds in the present invention by means of the prior art and the methods described in the present invention, so as to obtain chemically stable and easy-to-synthesize compounds.

本发明所述化合物可能含有一个或多个不对称中心，并可能由此产生非对映异构体和光学异构体。本发明包括所有可能的非对映异构体及其外消旋混合物、其基本上纯的拆分对映异构体、所有可能的几何异构体及其药学上可接受的盐。The compounds of the present invention may contain one or more asymmetric centers and may thereby give rise to diastereoisomers and optical isomers. The present invention includes all possible diastereoisomers and their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers and their pharmaceutically acceptable salts.

当式(I)所示化合物存在互变异构体时，除非特别声明，本发明包括任何可能的互变 异构体和其药学上可接受的盐，及它们的混合物。When the compounds represented by formula (I) have tautomers, unless otherwise stated, the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof.

当式(I)所示化合物用较重的同位素(例如氘)替代可能提供某些治疗优势，这是由于更大的代谢稳定性，例如增加体内半衰期或减少剂量要求。Substitution of compounds of formula (I) with heavier isotopes (eg deuterium) may afford certain therapeutic advantages due to greater metabolic stability, eg increased in vivo half-life or reduced dosage requirements.

当式(I)所示化合物及其药学上可接受的盐存在溶剂化物或多晶型时，本发明包括任何可能的溶剂化物和多晶型。形成溶剂化物的溶剂类型没有特别的限定，只要该溶剂是药学上可以接受的。例如，水、乙醇、丙醇、丙酮等类似的溶剂都可以采用。When the compound represented by formula (I) and its pharmaceutically acceptable salts have solvates or polymorphs, the present invention includes any possible solvates and polymorphs. The type of solvent forming a solvate is not particularly limited as long as the solvent is pharmaceutically acceptable. For example, water, ethanol, propanol, acetone, and the like can be used.

术语“组合物”，在本发明中，是指包括包含指定量的各指定成分的产品，以及直接或间接地由指定量的各指定成分的组合生产的任何产品。因此，含有本发明的化合物作为活性成分的药物组合物以及制备本发明化合物的方法也是本发明的一部分。此外，化合物的一些结晶形式可以多晶型存在，并且此多晶型包括在本发明中。另外，一些化合物可以与水(即水合物)或常见的有机溶剂形成溶剂化物，并且此类溶剂化物也落入本发明的范围内。The term "composition", in the present invention, is meant to include products comprising the specified amounts of each of the specified ingredients, as well as any product produced directly or indirectly from the combination of the specified amounts of each of the specified ingredients. Accordingly, pharmaceutical compositions containing the compounds of the invention as active ingredients as well as processes for the preparation of the compounds of the invention are also part of the invention. Furthermore, some of the crystalline forms of the compounds may exist as polymorphs, and such polymorphs are included in the present invention. In addition, some of the compounds may form solvates with water (ie, hydrates) or common organic solvents, and such solvates are also within the scope of the present invention.

本发明提供的药物组合物包括作为活性组分的式(I)所示化合物(或其药学上可接受的盐)、一种药学上可接受的赋形剂及其他可选的治疗组分或辅料。尽管任何给定的情况下，最适合的活性组分给药方式取决于接受给药的特定的主体、主体性质和病情严重程度。本发明的药物组合物可以方便地以本领域公知的单位剂型存在和药学领域公知的任何制备方法制备。The pharmaceutical composition provided by the present invention comprises a compound represented by formula (I) (or a pharmaceutically acceptable salt thereof) as an active component, a pharmaceutically acceptable excipient and other optional therapeutic components or Accessories. In any given case though, the most suitable mode of administration of the active ingredient will depend upon the particular subject to be administered, the nature of the subject and the severity of the condition. The pharmaceutical compositions of the present invention may conveniently be presented in unit dosage forms and prepared by any methods of preparation well known in the art of pharmacy.

本发明的药物组合物包括药学上可接受的载体和式(I)所示化合物或其立体异构体、互变异构体，多晶型物、溶剂化物、其药学上可接受的盐、其药物前体。式(I)所示化合物或其药学上可接受的盐，与其他一种或多种具有治疗活性的化合物的联合用药也包括在本发明的药物组合物中。The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier and a compound represented by formula (I) or its stereoisomers, tautomers, polymorphs, solvates, pharmaceutically acceptable salts thereof, its drug prodrugs. The combination of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof and one or more other therapeutically active compounds is also included in the pharmaceutical composition of the present invention.

本发明的药物组合物包括适于口腔、直肠、局部和不经肠道(包括皮下给药、肌肉注射、静脉给药)给药的药物组合物。含有本发明化合物或药物组合物的片剂或胶囊可含有一种或多种辅助组分。较优地，每个片剂或胶囊含有大约0.05mg到5g的活性组分。本发明还提供适用于注射的药物组合物，包括无菌水溶液或分散体系。进一步地，上述药物组合物可以制备成无菌粉末形式以用于即时配制无菌注射液或分散液。本发明提供的药物组合物可以是适于局部用药的形式，例如，气溶胶、乳剂、软膏、洗液、撒粉或其他类似的剂型。进一步地，本发明提供的药物组合物可以采用适于经皮给药设备使用的形式。本发明提供的药物组合物，可以以固体为载体，适用于直肠给药的形式。单位剂量的栓剂是最典型的剂型。栓剂可以方便地制备，首先药物组合物与软化或熔化的辅料混合，然后冷却 和模具成型而制得。The pharmaceutical compositions of the present invention include pharmaceutical compositions suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular, and intravenous) administration. Tablets or capsules containing a compound or pharmaceutical composition of this invention may contain one or more accessory ingredients. Preferably, each tablet or capsule contains from about 0.05 mg to 5 g of active ingredient. The present invention also provides pharmaceutical compositions suitable for injection, including sterile aqueous solutions or dispersions. Furthermore, the above pharmaceutical composition can be prepared in the form of sterile powder for immediate preparation of sterile injection or dispersion. The pharmaceutical composition provided by the present invention can be in a form suitable for topical administration, for example, aerosol, cream, ointment, lotion, dusting powder or other similar dosage forms. Furthermore, the pharmaceutical composition provided by the present invention can be in a form suitable for use in transdermal drug delivery devices. The pharmaceutical composition provided by the invention may be in a form suitable for rectal administration with solid as carrier. Unit dose suppositories are the most typical dosage form. Suppositories are conveniently prepared by first mixing the pharmaceutical composition with softened or melted excipients, followed by cooling and moulding.

上述制剂配方还可以包括，适当的，一种或多种附加的辅料组分，如稀释剂、缓冲剂、调味剂、粘合剂、表面活性剂、增稠剂、润滑剂和防腐剂(包括抗氧化剂)等。进一步地，其他的辅药还可以包括调节药物与血液等渗压的促渗剂。包含式(I)所示化合物，或其药学上可接受的盐的药物组合物，可以制备成粉剂或浓缩液的形式。The formulation of the above preparation may also include, as appropriate, one or more additional adjuvant components, such as diluents, buffers, flavoring agents, binding agents, surfactants, thickeners, lubricants and preservatives (including antioxidants), etc. Further, other adjuvants may also include penetration enhancers that adjust the isotonic pressure between the drug and the blood. The pharmaceutical composition comprising the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, can be prepared in the form of powder or concentrated solution.

一般情况下，治疗上述所示的状况或不适，药物的剂量水平约为每天0.01mg/kg体重到150mg/kg体重，或者每个病人每天0.5mg到7g。但是，可以理解，可能需要比上述那些更低或更高的剂量。任何特定病人的具体剂量水平和治疗方案将取决于多种因素，包括所用具体化合物的活性、年龄、体重、综合健康状况、性别、饮食、给药时间、给药途径、排泄率、药物联用的情况和接受治疗的特定疾病的严重程度。Generally, for the treatment of the conditions or disorders indicated above, the dosage level of the drug is about 0.01 mg/kg body weight to 150 mg/kg body weight per day, or 0.5 mg to 7 g per day per patient. However, it will be appreciated that lower or higher dosages than those recited above may be required. The specific dosage level and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the particular compound employed, age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combinations condition and the severity of the particular disease being treated.

附图说明Description of drawings

图1为实施例1A制得化合物1a的X射线单晶衍射谱图A。Fig. 1 is the X-ray single crystal diffraction pattern A ofcompound 1a prepared in Example 1A.

图2为实施例1A制得化合物1a的X射线单晶衍射谱图B。Fig. 2 is the X-ray single crystal diffraction spectrum B ofcompound 1a prepared in Example 1A.

图3为实施例1A制得化合物1a的绝对构型图。Figure 3 is the absolute configuration diagram ofcompound 1a prepared in Example 1A.

图4为人胰腺癌MIA PaCa-2 CDX肿瘤模型上的体内药效学研究A。Figure 4 is the in vivo pharmacodynamic study A on the human pancreatic cancer MIA PaCa-2 CDX tumor model.

图5为人胰腺癌MIA PaCa-2 CDX肿瘤模型上的体内药效学研究B。Figure 5 is the in vivo pharmacodynamic study B on the human pancreatic cancer MIA PaCa-2 CDX tumor model.

图6为人肺癌PDX肿瘤模型的体内药效学研究。Fig. 6 is an in vivo pharmacodynamic study of a human lung cancer PDX tumor model.

具体实施方式Detailed ways

为使上述内容更清楚、明确，本发明将用以下实施例来进一步阐述本发明的技术方案。以下实施例仅用于说明本发明的具体实施方式，以使本领域的技术人员能够理解本发明，但不用于限制本发明的保护范围。本发明的具体实施方式中，未作特别说明的技术手段或方法等为本领域的常规技术手段或方法等。In order to make the above content more clear and definite, the present invention will use the following examples to further illustrate the technical solution of the present invention. The following examples are only used to illustrate specific embodiments of the present invention so that those skilled in the art can understand the present invention, but are not intended to limit the protection scope of the present invention. In the specific implementation of the present invention, technical means or methods that are not specifically described are conventional technical means or methods in the art.

除非另有说明，本发明所有的一部分和百分比均按重量计算，所有温度均指摄氏度。实施例中使用了下列缩略语：Unless otherwise indicated, all parts and percentages herein are by weight and all temperatures are in degrees Celsius. The following abbreviations are used in the examples:

BOP：卡特缩合剂；BOP: Carter condensing agent;

CDI：羰基二咪唑；CDI: carbonyldiimidazole;

DBU:1,8-二氮杂双环[5.4.0]十一碳-7-烯；DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene;

DIEA：N，N-二异丙基乙胺；DIEA: N,N-Diisopropylethylamine;

DMF：N，N-二甲基甲酰胺；DMF: N, N-dimethylformamide;

DCM:二氯甲烷；DCM: dichloromethane;

Dioxane：二氧六环；Dioxane: dioxane;

ESI-MS：电喷雾电离质谱；ESI-MS: electrospray ionization mass spectrometry;

EtOH：乙醇；EtOH: ethanol;

HOAc：冰醋酸；HOAc: glacial acetic acid;

MeOH：甲醇；MeOH: Methanol;

NIS：N-碘代丁二酰亚胺；NIS: N-iodosuccinimide;

NCS：N-氯代丁二酰亚胺；NCS: N-chlorosuccinimide;

PE:EA：石油醚和乙酸乙酯的比值；PE:EA: ratio of petroleum ether to ethyl acetate;

POCl3：三氯氧磷；POCl3 : phosphorus oxychloride;

SOCl2：二氯亚砜；SOCl2 : thionyl chloride;

THF：四氢呋喃；THF: Tetrahydrofuran;

TFA：三氟乙酸；TFA: trifluoroacetic acid;

TEA：三乙胺；TEA: triethylamine;

Toluene：甲苯；Toluene: toluene;

Sphos Pd G2：氯(2-二环己基膦基-2,6-二甲氧基-1,1-联苯基)(2-氨基-1,1-联苯-2-基)钯(II)；Sphos Pd G2: Chloro(2-dicyclohexylphosphino-2,6-dimethoxy-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II );

Pd(dppf)Cl2.CH2Cl2：[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物；Pd(dppf)Cl2 .CH2 Cl2 : [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex;

Pd(PPh3)4：四(三苯基膦)钯；Pd(PPh3 )4 : tetrakis(triphenylphosphine) palladium;

Pre-TLC:薄层层析硅胶板。Pre-TLC: Thin layer chromatography silica gel plate.

中间体化合物M1合成Synthesis of intermediate compound M1

Figure PCTCN2021124884-APPB-000034
Figure PCTCN2021124884-APPB-000034

步骤1：化合物M1-2的合成Step 1: Synthesis of compound M1-2

在室温下，向化合物M1-1(40g)，HOAc(76.8g)，EtOH(400mL)与H2O(160mL)的混合物中逐份添加铁粉(26.52g)。所得混合物在室温下搅拌2小时，随后用NaOH(5N)溶液中和。随后用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩得褐色油状的所要粗品产物(34g，98％产率)，即化合物M1-2。To a mixture of compound M1-1 (40 g), HOAc (76.8 g), EtOH (400 mL) and H2 O (160 mL) was added iron powder (26.52 g) in portions at room temperature. The resulting mixture was stirred at room temperature for 2 hours, then neutralized with NaOH (5N) solution. The mixture was then extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to afford the desired crude product (34 g, 98% yield), compound M1-2, as a brown oil.

ESI-MS m/z:190.02[M+H]+。ESI-MS m/z: 190.02 [M+H]+ .

步骤2：化合物M1-3的合成Step 2: Synthesis of compound M1-3

将2,2,2-三氯乙烷-1,1-二醇(66.4g)与Na2SO4(503.4g)溶于水(560mL)中，然后升温热至55℃。添加含有化合物M1-2(34g)的水(240mL)和35％HC1(72mL)，再添加盐酸羟胺(81.4g)的水溶液(100mL)。所得混合物在90℃搅拌3小时且形成黄色沉淀物。将混合物冷却至室温。通过过滤收集固体,用水冲洗,且风干,得到黄褐色固体产物(47g，99％产率)，即化合物M1-3。2,2,2-Trichloroethane-1,1-diol (66.4g) and Na2 SO4 (503.4g) were dissolved in water (560mL), and then heated to 55°C. Water (240 mL) and 35% HCl (72 mL) containing compound M1-2 (34 g) were added, and an aqueous solution (100 mL) of hydroxylamine hydrochloride (81.4 g) was added. The resulting mixture was stirred at 90 °C for 3 hours and a yellow precipitate formed. The mixture was cooled to room temperature. The solid was collected by filtration, rinsed with water, and air-dried to give the product as a tan solid (47 g, 99% yield), compound M1-3.

ESI-MS m/z:261.03[M+H]+。ESI-MS m/z: 261.03 [M+H]+ .

步骤3：化合物M1-4的合成Step 3: Synthesis of compound M1-4

在60℃，向浓硫酸(300mL)中添加化合物M1-3(47g),将温度升高至90℃且维持3小时，反应完全，将反应混合物冷却至室温且倾注入冰水中。通过过滤收集黄色沉淀物且干燥，得到黑色固体产物(43g，99％产率)，即化合物M1-4。Compound M1-3 (47 g) was added to concentrated sulfuric acid (300 mL) at 60 °C, the temperature was raised to 90 °C and maintained for 3 hours, the reaction was complete, the reaction mixture was cooled to room temperature and poured into ice water. The yellow precipitate was collected by filtration and dried to give a black solid product (43 g, 99% yield), compound M1-4.

步骤4：化合物M1-5的合成Step 4: Synthesis of Compound M1-5

在0℃，将化合物M1-4(43g)于NaOH(2N,500mL)中的溶液中添加H2O2溶液(30％，80mL)且所得混合物在0℃搅拌30分钟。再移至室温下搅拌2小时，反应完全，将混合物倾注入冰水中再用浓HCI溶液酸化,通过过滤收集沉淀物且风干，得到呈白色固体状产物(20g，48.9％产率)，即化合物M1-5。To a solution of compound M1-4 (43 g) in NaOH (2N, 500 mL) was added H2 O2 solution (30%, 80 mL) at 0° C. and the resulting mixture was stirred at 0° C. for 30 min. Then moved to room temperature and stirred for 2 hours, the reaction was complete, the mixture was poured into ice water and acidified with concentrated HCI solution, the precipitate was collected by filtration and air-dried to obtain the product (20 g, 48.9% yield) as a white solid, namely compound M1-5.

ESI-MS m/z:233.97[M+H]+。ESI-MS m/z: 233.97 [M+H]+ .

步骤5：化合物M1-6的合成Step 5: Synthesis of compound M1-6

在室温下，向化合物M1-5(20g)于DMF(200mL)中的溶液中添加NIS(29g)且所得混合物在70℃搅拌过夜。反应完全，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩得褐色固体状的所要粗品产物(30g，98％产率)，即化合物M1-6。To a solution of compound M1-5 (20 g) in DMF (200 mL) was added NIS (29 g) at room temperature and the resulting mixture was stirred at 70°C overnight. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried overNa2SO4 and concentrated in vacuo to give the desired crude product (30g , 98% yield) as a brown solid, i.e. Compound M1-6.

ESI-MS m/z:359.87[M+H]+。ESI-MS m/z: 359.87 [M+H]+ .

1H NMR(500MHz,DMSO)δ13.34(s,1H),7.99(s,1H),6.87(s,2H)。1H NMR (500MHz, DMSO) δ 13.34(s, 1H), 7.99(s, 1H), 6.87(s, 2H).

步骤6：化合物M1的合成Step 6: Synthesis of Compound M1

在室温下，将二(咪唑-1-基)甲酮(2.70g)加入到粗品化合物M1-6(4.0g)的THF(20mL)中，再将N-乙基-N-异丙基丙-2-胺(1.44g，1.94mL)加入其中，混合物移至50℃反应，约反应2小时化合物M1-6基本完全转化为中间产物，接着将混合物逐滴加入到冰的氨水(35mL)中，搅拌5min即反应完全。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(石油醚/乙酸乙酯＝70:30)纯化，得到褐色固体状的所要目标产物化合物M1(1.64g)。At room temperature, bis(imidazol-1-yl)methanone (2.70g) was added to crude compound M1-6 (4.0g) in THF (20mL), and N-ethyl-N-isopropylpropyl -2-Amine (1.44g, 1.94mL) was added thereto, the mixture was moved to 50°C for reaction, and the compound M1-6 was almost completely converted into an intermediate product after about 2 hours of reaction, and then the mixture was added dropwise to iced ammonia water (35mL) , and stirred for 5 minutes to complete the reaction. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried overNa2SO4 and concentrated in vacuo, the residue was purified by flashsilica gel column chromatography (petroleum ether/ethyl acetate=70:30) , the desired target compound M1 (1.64 g) was obtained as a brown solid.

中间体化合物M2合成Synthesis of intermediate compound M2

Figure PCTCN2021124884-APPB-000035
Figure PCTCN2021124884-APPB-000035

步骤1：化合物M2-1的合成Step 1: Synthesis of Compound M2-1

在室温条件下，将化合物M1-5(3.6g)溶于DMF(20mL)中，加入NCS(2.05g)，在70℃反应1.5h。反应完全，降到室温，加水稀释，用EA萃取三次，合并有机相并用无水Na2SO4干燥，过滤，旋干得棕色固体M2-1(3.86g，粗品)。Compound M1-5 (3.6 g) was dissolved in DMF (20 mL) at room temperature, NCS (2.05 g) was added, and reacted at 70° C. for 1.5 h. The reaction was complete, cooled to room temperature, diluted with water, extracted three times with EA, combined the organic phases and dried with anhydrous Na2 SO4 , filtered, and spin-dried to obtain a brown solid M2-1 (3.86 g, crude product).

ESI-MS m/z:267.91[M+H]+。ESI-MS m/z: 267.91 [M+H]+ .

步骤2：化合物M2-2的合成Step 2: Synthesis of compound M2-2

在室温条件下，将化合物M2-1(3.7g)溶于THF(20mL)中，依次加入CDI(2.98g，)，DIEA(5.34g)，在50℃反应1.5h后，在冰浴条件下将反应液逐滴到氨水(40mL)中，搅拌5min即反应完全。加水稀释，用EA萃取三次，合并有机相并用无水Na2SO4干燥，过滤，旋干。硅胶柱分离纯化(PE:EA＝2:1)得黄色固体M2-2(2.32g，62.93％产率)。At room temperature, compound M2-1 (3.7g) was dissolved in THF (20mL), and CDI (2.98g, ) and DIEA (5.34g) were added successively, and reacted at 50°C for 1.5h. The reaction solution was added dropwise into aqueous ammonia (40 mL), and stirred for 5 min to complete the reaction. Diluted with water, extracted threetimes with EA, combined the organic phases and dried over anhydrousNa2SO4 , filtered and spin-dried. Silica gel column separation and purification (PE:EA=2:1) gave yellow solid M2-2 (2.32 g, 62.93% yield).

ESI-MS m/z:266.93[M+H]+。ESI-MS m/z: 266.93 [M+H]+ .

步骤3：化合物M2-3的合成Step 3: Synthesis of compound M2-3

在室温条件下，将化合物1-1(4.17g)溶于THF(10mL)中，加入M2-2(2.3g)，40℃反应4h。降到室温，加水淬灭，用EA萃取三次，合并有机相并用无水硫酸钠干燥，过滤，旋干。得棕色固体M2-3(3.4g，粗品)。At room temperature, compound 1-1 (4.17 g) was dissolved in THF (10 mL), M2-2 (2.3 g) was added, and reacted at 40° C. for 4 h. Cool down to room temperature, add water to quench, extract three times with EA, combine organic phases and dry over anhydrous sodium sulfate, filter and spin dry. A brown solid M2-3 (3.4 g, crude) was obtained.

ESI-MS m/z:392.01[M+H]+。ESI-MS m/z: 392.01 [M+H]+ .

步骤4：化合物M2-4的合成Step 4: Synthesis of compound M2-4

在室温条件下，将甲醇钠(2.34g)加入到化合物M2-3(3.4g)的甲苯(50mL)中，移至110℃回流搅拌5小时。降到室温，加水稀释，用3N HCl调节pH至6，用EA萃取三次，合并有机相并用无水Na2SO4干燥，过滤，旋干，得到棕色固体M2-4(2.63g，粗品)。At room temperature, sodium methoxide (2.34 g) was added to compound M2-3 (3.4 g) in toluene (50 mL), and the mixture was moved to 110° C. under reflux and stirred for 5 hours. Cooled down to room temperature, diluted with water, adjusted pH to 6 with 3N HCl, extracted three times with EA, combined organic phases and dried over anhydrous Na2 SO4 , filtered, and spin-dried to give brown solid M2-4 (2.63 g, crude).

ESI-MS m/z:374.01[M+H]+。ESI-MS m/z: 374.01 [M+H]+ .

步骤5：化合物M2-5的合成Step 5: Synthesis of compound M2-5

在室温条件下，氮气保护下，将DIEA(5mL)加入到化合物M2-4(2.6g)的POCl3(50mL)中，移至110℃搅拌2小时。待反应完全后，直接浓缩除去POCl3，得到棕褐色固体M2-5(2.8g，粗品)。At room temperature, under nitrogen protection, DIEA (5 mL) was added into POCl3 (50 mL) of compound M2-4 (2.6 g), moved to 110° C. and stirred for 2 hours. After the reaction was complete, the reaction was directly concentrated to remove POCl3 to obtain tan solid M2-5 (2.8 g, crude product).

ESI-MS m/z:391.97[M+H]+。ESI-MS m/z: 391.97 [M+H]+ .

步骤6：化合物M2的合成Step 6: Synthesis of compound M2

在室温条件下，将DIEA(4.60g)加入到化合物M2-5(2.8g)，2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(1.93g)的二氧六环(30mL)中，室温反应5min。加水淬灭，用EA萃取三次，合并有机相并用无水Na2SO4干燥，过滤，旋干，硅胶柱分离纯化(DCM:MeOH＝10:1)，得到棕色固体M2(3.4g，81.9％产率)。At room temperature, DIEA (4.60g) was added to compound M2-5 (2.8g), 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (1.93g) in dioxygen Hexacyclic (30 mL), react at room temperature for 5 min. Quenched with water, extracted three times with EA, combined the organic phases and dried with anhydrous Na2 SO4 , filtered, spin-dried, and purified by silica gel column separation (DCM:MeOH=10:1) to obtain a brown solid M2 (3.4g, 81.9% Yield).

ESI-MS m/z:582.16[M+H]+。ESI-MS m/z: 582.16 [M+H]+ .

实施例1：化合物1(1-(7-(7-(5-甲基-1H-吲唑-4-基)-2-(1甲基哌啶-4-基)-8-(2，2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 1: Compound 1 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(1 methylpiperidin-4-yl)-8-(2, 2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one )Synthesis

Figure PCTCN2021124884-APPB-000036
Figure PCTCN2021124884-APPB-000036

步骤1：化合物1-1的合成Step 1: Synthesis of compound 1-1

在室温下，氮气保护下，将1-甲基哌啶-4-羧酸(500mg)溶于SOCl2(2mL)中，移至70℃下，搅拌反应1h。真空浓缩，除去氯化亚砜，得到白色固体状目标产物，即化合物1-1(520mg，粗品)。At room temperature, under nitrogen protection, 1-methylpiperidine-4-carboxylic acid (500 mg) was dissolved in SOCl2 (2 mL), moved to 70° C., and stirred for 1 h. Concentrate in vacuo to remove thionyl chloride to obtain the target product, compound 1-1 (520 mg, crude product) as a white solid.

步骤2：化合物1-2的合成Step 2: Synthesis of Compound 1-2

在室温下，将化合物1-1(405.27mg)加入到化合物M1(450mg)的THF(10mL)中，移至40℃搅拌4小时。待反应完全后，直接浓缩出去THF，得到白色固体状粗品目标产物，即化合物1-2(600mg，粗品)。Compound 1-1 (405.27 mg) was added to compound M1 (450 mg) in THF (10 mL) at room temperature, and the mixture was moved to 40° C. and stirred for 4 hours. After the reaction was complete, the THF was directly concentrated to obtain the crude target product, compound 1-2 (600 mg, crude product) as a white solid.

ESI-MS m/z:483.94[M+H]+。ESI-MS m/z: 483.94 [M+H]+ .

步骤3：化合物1-3的合成Step 3: Synthesis of compounds 1-3

在室温下，将甲醇钠(200.87mg)加入到化合物1-2(600mg)的甲苯(15mL)中，移至110℃回流搅拌5小时。待反应完全后，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机相经Na2SO4干燥且真空浓缩，得到白色固体状粗品目标产物，即化合物1-3(600mg，粗品)。Sodium methoxide (200.87 mg) was added to compound 1-2 (600 mg) in toluene (15 mL) at room temperature, and the mixture was moved to 110° C. under reflux and stirred for 5 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo to obtain the crude target product, compound 1-3 (600 mg, crude) as a white solid.

ESI-MS m/z:465.96[M+H]+。ESI-MS m/z: 465.96 [M+H]+ .

步骤4：化合物1-4的合成Step 4: Synthesis of compounds 1-4

在室温下，氮气保护下，将DIEA(0.5mL)加入到化合物1-3(600mg)的POCl3(5mL)中，移至110℃搅拌3小时。待反应完全后，直接浓缩除去POCl3，得到棕褐色固体状粗品目标产物，即化合物1-4(600mg，粗品)。At room temperature, under nitrogen protection, DIEA (0.5 mL) was added to POCl3 (5 mL) of compound 1-3 (600 mg), moved to 110° C. and stirred for 3 hours. After the reaction was complete, the mixture was directly concentrated to remove POCl3 to obtain the crude target product, compound 1-4 (600 mg, crude product) as a tan solid.

ESI-MS m/z:483.96[M+H]+。ESI-MS m/z: 483.96 [M+H]+ .

步骤5：化合物1-5的合成Step 5: Synthesis of Compounds 1-5

在室温下，将DIEA(480.12mg)加入到化合物1-4(600mg)，2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(280.25mg)的1,4-二氧六环(10mL)中，室温搅拌3小时。将混合物倾注入冰水中，用乙酸乙酯/甲醇＝10:1萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(DCM:MeOH＝10:1)纯化，得到黄色固体状的所要目标产物，即化合物1-5(750mg，89.81％产率)。At room temperature, DIEA (480.12mg) was added to compound 1-4 (600mg), 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (280.25mg) in 1,4- Dioxane (10 mL), stirred at room temperature for 3 hours. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (DCM:MeOH=10: 1) Purification to obtain the desired target product, compound 1-5 (750 mg, 89.81% yield) as a yellow solid.

ESI-MS m/z:674.10[M+H]+。ESI-MS m/z: 674.10 [M+H]+ .

步骤6：化合物1-6的合成Step 6: Synthesis of Compounds 1-6

在氮气保护下，将三氟乙醇(244.76mg)加入到化合物1-5(550mg)，Cs2CO3(531.45mg，)的1,4-二氧六环(5mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯/甲醇＝10:1萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物，即化合物1-6(590mg，粗品)。Under nitrogen protection, trifluoroethanol (244.76mg) was added to compound 1-5 (550mg), Cs2 CO3 (531.45mg,) in 1,4-dioxane (5mL), and moved to 100°C React for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product, compound 1-6, as a yellow solid (590 mg, crude).

ESI-MS m/z:755.30[M+H]+。ESI-MS m/z: 755.30 [M+H]+ .

步骤7：化合物1-7的合成Step 7: Synthesis of Compounds 1-7

在氮气保护下，将Pd(dppf)Cl2.CH2Cl2(63.82mg)加入到化合物1-6(590mg)，K2CO3(216.17mg)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(120.45mg)的1,4-二氧六环(5.0mL)，H2O(0.5mL)混合溶液中，在70℃搅拌1h。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化，得到呈黄色固体状的所要产物，即化合物1-7(329mg，64.27％产率)。Under nitrogen protection, Pd(dppf)Cl2 .CH2 Cl2 (63.82 mg) was added to compound 1-6 (590 mg), K2 CO3 (216.17 mg), 4,4,5,5-tetramethyl In a mixed solution of 2-ethenyl-1,3,2-dioxaborolane (120.45 mg) in 1,4-dioxane (5.0 mL), H2 O (0.5 mL), Stir at 70 °C for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1) to obtain the desired product, compound 1-7 (329 mg, 64.27% yield) as a yellow solid.

ESI-MS m/z:655.63[M+H]+。ESI-MS m/z: 655.63 [M+H]+ .

步骤8：化合物1-8的合成Step 8: Synthesis of Compounds 1-8

在氮气保护下，将Pd(PPh3)4(116.16mg)加入到化合物1-7(329mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(344.04mg)，K3PO4(213.38mg，)的 1,4-二氧六环(3mL)，H2O(0.75mL)溶液中，移至85℃反应反应约3小时。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化，得到呈黄色固体状的所要产物，即化合物1-8(244mg，61.45％产率)。Under nitrogen protection, Pd(PPh3 )4 (116.16mg) was added to compound 1-7 (329mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (344.04mg), K3 PO4 (213.38mg,) of 1,4-dioxane (3mL ), H2 O (0.75 mL) solution, moved to 85° C. for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1) to obtain the desired product, compound 1-8 (244 mg, 61.45% yield) as a yellow solid.

ESI-MS m/z:874.10[M+H]+。ESI-MS m/z: 874.10 [M+H]+ .

步骤9：化合物1-9的合成Step 9: Synthesis of Compounds 1-9

在室温下，将化合物1-8(244mg)溶于DCM(4mL)和TFA(2mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色固体状粗品目标产物，即化合物1-9(190mg，粗品)。Compound 1-8 (244 mg) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product, Compound 1-9 (190 mg, crude product) as a yellow solid.

ESI-MS m/z:606.38[M+H]+。ESI-MS m/z: 606.38 [M+H]+ .

步骤10：化合物1的合成Step 10: Synthesis ofcompound 1

在氮气保护下，冰水浴中，将丙烯酰氯(17.03mg)的THF溶液加入到化合物1-9(95mg)，THF(4mL)，饱和Na2CO3(1mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物，即化合物1(24.1mg，22.96％产率)。Under nitrogen protection, a THF solution of acryloyl chloride (17.03 mg) was added to compound 1-9 (95 mg), THF (4 mL), saturated Na2 CO3 (1 mL) solution in an ice-water bath, and the reaction was complete upon addition. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product, Compound 1 (24.1 mg, 22.96% yield) as a yellow solid.

ESI-MS m/z:660.37[M+H]+。ESI-MS m/z: 660.37 [M+H]+.

1H NMR(500MHz,Methanol-d4)δ8.08(s,1H),7.52(d,J＝8.6Hz,1H),7.43–7.32(m,2H),6.40(d,J＝17.0,10.3Hz,1H),6.32–6.17(m,2H),5.78–5.69(m,2H),5.09(d,J＝11.1Hz,1H),4.86–4.75(m,1H),4.51(m,1H),4.15(s,2H),3.90(s,2H),3.72(td,J＝6.4,5.2,3.0Hz,4H),3.02(d,J＝11.4Hz,2H),2.85(t,J＝11.5Hz,1H),2.34(s,3H),2.25(m,2H),2.13(m,5H),2.06(m,3H),1.90–1.83(m,4H).1 H NMR (500MHz, Methanol-d4 )δ8.08(s,1H),7.52(d,J=8.6Hz,1H),7.43–7.32(m,2H),6.40(d,J=17.0,10.3 Hz,1H),6.32–6.17(m,2H),5.78–5.69(m,2H),5.09(d,J＝11.1Hz,1H),4.86–4.75(m,1H),4.51(m,1H) ,4.15(s,2H),3.90(s,2H),3.72(td,J=6.4,5.2,3.0Hz,4H),3.02(d,J=11.4Hz,2H),2.85(t,J=11.5 Hz,1H),2.34(s,3H),2.25(m,2H),2.13(m,5H),2.06(m,3H),1.90–1.83(m,4H).

实施例1A：化合物1的拆分与异构体结构解析Example 1A: Resolution and Isomer Structure Analysis ofCompound 1

使用色谱条件：CHIRALCEL OD(2.5cm I.D.×25cm L,10μm)，UV 254nm，流动相Hexane/IPA/DEA＝70/30/0.1(V/V/V)，流速20ml/min，拆分92mg化合物1，获得异构体1a(56.3mg)和异构体1b(34.3mg)。Chromatographic conditions used: CHIRALCEL OD (2.5cm I.D.×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, resolve92mg compound 1.Isomer 1a (56.3 mg) and Isomer 1b (34.3 mg) were obtained.

Figure PCTCN2021124884-APPB-000037
Figure PCTCN2021124884-APPB-000037

异构体1a：1H NMR(500MHz,DMSO-d6)δ13.00(s,1H),8.00(s,1H),7.49(d,J＝8.5Hz,1H),7.40(s,1H),7.32(d,J＝8.6Hz,1H),6.35(dd,J＝17.0,10.3Hz,1H),6.18–6.06(m,2H),5.79–5.66(m,2H),5.11(d,J＝11.1Hz,1H),4.97(dq,J＝12.3,9.1Hz,1H),4.68(dq,J＝12.3,9.1Hz,1H),4.05(s,2H),3.81–3.72(m,6H),2.86(dt,J＝11.5,3.1Hz,2H),2.68(tt,J＝11.4,3.6Hz,1H),2.19(s,3H),2.04(s,3H),1.98(dt,J＝6.7,4.1Hz,8H),1.85(qd,J＝13.0,3.9Hz,2H).ESI-MS m/z:660.4[M+H]+。色谱条件：CHIRALCEL OD(2.5cm I.D.×25cm L,10μm)，UV 254nm，流动相Hexane/IPA/DEA＝70/30/0.1(V/V/V)，流速20ml/min，保留时间6.39min(后出峰)；Isomer 1a:1 H NMR (500MHz, DMSO-d6 ) δ13.00(s, 1H), 8.00(s, 1H), 7.49(d, J=8.5Hz, 1H), 7.40(s, 1H) ,7.32(d,J=8.6Hz,1H),6.35(dd,J=17.0,10.3Hz,1H),6.18–6.06(m,2H),5.79–5.66(m,2H),5.11(d,J =11.1Hz,1H),4.97(dq,J=12.3,9.1Hz,1H),4.68(dq,J=12.3,9.1Hz,1H),4.05(s,2H),3.81–3.72(m,6H) ,2.86(dt,J=11.5,3.1Hz,2H),2.68(tt,J=11.4,3.6Hz,1H),2.19(s,3H),2.04(s,3H),1.98(dt,J=6.7 , 4.1Hz, 8H), 1.85 (qd, J=13.0, 3.9Hz, 2H). ESI-MS m/z: 660.4[M+H]+. Chromatographic conditions: CHIRALCEL OD (2.5cm ID×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, retention time 6.39min ( after the peak);

异构体1b：1H NMR(500MHz,DMSO-d6)δ13.01(s,1H),8.00(s,1H),7.50(d,J＝8.5Hz,1H),7.41(s,1H),7.32(d,J＝8.6Hz,1H),6.35(dd,J＝17.0,10.3Hz,1H),6.19–6.06(m,2H),5.80–5.60(m,2H),5.11(d,J＝11.2Hz,1H),4.98(dq,J＝12.2,9.1Hz,1H),4.69(dq,J＝12.2,9.2Hz,1H),4.05(s,2H),3.76(d,J＝7.1Hz,6H),2.85(dd,J＝8.9,5.8Hz,2H),2.68(tt,J＝11.6,3.7Hz,1H),2.19(s,3H),2.05(s,3H),2.02–1.96(m,8H),1.85(qd,J＝13.2,4.0Hz,2H).ESI-MS m/z:660.4[M+H]+。色谱条件：CHIRALCEL OD(2.5cm I.D.×25cm L,10μm)，UV 254nm，流动相Hexane/IPA/DEA＝70/30/0.1(V/V/V)，流速20ml/min，保留时间4.09min(先出峰)；Isomer 1b:1 H NMR (500MHz, DMSO-d6 ) δ13.01(s, 1H), 8.00(s, 1H), 7.50(d, J=8.5Hz, 1H), 7.41(s, 1H) ,7.32(d,J=8.6Hz,1H),6.35(dd,J=17.0,10.3Hz,1H),6.19–6.06(m,2H),5.80–5.60(m,2H),5.11(d,J =11.2Hz,1H),4.98(dq,J=12.2,9.1Hz,1H),4.69(dq,J=12.2,9.2Hz,1H),4.05(s,2H),3.76(d,J=7.1Hz ,6H),2.85(dd,J＝8.9,5.8Hz,2H),2.68(tt,J＝11.6,3.7Hz,1H),2.19(s,3H),2.05(s,3H),2.02–1.96( m, 8H), 1.85 (qd, J = 13.2, 4.0Hz, 2H). ESI-MS m/z: 660.4 [M+H]+. Chromatographic conditions: CHIRALCEL OD (2.5cm ID×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, retention time 4.09min ( first out of the peak);

通过在DMSO/H2O体系液面扩散方法进行单晶培养，得到异构体1a的DMSO溶剂合物的单晶。晶体衍射实验用晶体尺寸为0.03×0.19×0.23mm，采用Bruker D8 Venture Photon II衍射仪进行单晶测试，光源为CuKα辐射，扫描方式为

Figure PCTCN2021124884-APPB-000038
扫描。采用直接法(Shelxs97)解析晶体结构，获得全部100个非氢原子位置，晶体属于单斜晶系，空间群为P21，晶胞参数：a＝18.4614(13)，b＝11.8865(8)，
Figure PCTCN2021124884-APPB-000039
α＝γ＝90°，β＝110.112(3)°，晶胞体积
Figure PCTCN2021124884-APPB-000040
晶胞内不对称单位数Z＝2。使用最小二乘法修正结构参数和判别原子种类，使用几何计算法和差值Fourier法获得全部氢原子位置。单晶结构解析结果表明：晶态下分子排列属第一类空间群，化合物应具有旋光活性，Flack系数0.044(12)，可确定晶体中化 合物的绝对构型为S构型。The single crystal of the DMSO solvate of theisomer 1a is obtained by culturing the single crystal by the liquid surface diffusion method in the DMSO/H2 O system. The crystal size used in the crystal diffraction experiment is 0.03×0.19×0.23mm, and the Bruker D8 Venture Photon II diffractometer is used for single crystal testing. The light source is CuKα radiation, and the scanning method is
Figure PCTCN2021124884-APPB-000038
scanning. The crystal structure was analyzed by the direct method (Shelxs97), and all 100 non-hydrogen atom positions were obtained. The crystal belongs to the monoclinic system, the space group is P21 , and the unit cell parameters: a=18.4614(13), b=11.8865(8),
Figure PCTCN2021124884-APPB-000039
α=γ=90°, β=110.112(3)°, unit cell volume
Figure PCTCN2021124884-APPB-000040
The number of asymmetric units in the unit cell is Z=2. The least squares method was used to modify the structural parameters and the atomic species were discriminated, and the geometric calculation method and the difference Fourier method were used to obtain the positions of all hydrogen atoms. The results of single crystal structure analysis show that the molecular arrangement in the crystal state belongs to the first space group, the compound should be optically active, and the Flack coefficient is 0.044(12), so the absolute configuration of the compound in the crystal can be determined as S configuration.

[根据细则91更正 15.02.2022]实施例2：化合物2(1-(7-(2-环己基-7-(5-甲基-1H-吲唑-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2，7-二氮杂螺[3.5]壬烷-2-基)丙-2-烯-1-酮)的合成[Corrected under Rule 91 15.02.2022]Example 2: Compound 2 (1-(7-(2-cyclohexyl-7-(5-methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy )-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]nonan-2-yl)prop-2-en-1-one) synthesis

Figure PCTCN2021124884-APPB-000041
Figure PCTCN2021124884-APPB-000041

步骤1：化合物2-1的合成Step 1: Synthesis of Compound 2-1

在室温下，将环己甲酰氯(367.63mg)加入到化合物M1(300mg)的THF(10mL)中，移至40℃搅拌4小时。待反应完全后，直接浓缩出去THF，得到白色固体状粗品目标产物，即化合物2-1(390mg，粗品)。Cyclohexanoyl chloride (367.63 mg) was added to compound M1 (300 mg) in THF (10 mL) at room temperature, and the mixture was moved to 40°C and stirred for 4 hours. After the reaction was complete, the THF was directly concentrated to obtain the crude target product, compound 2-1 (390 mg, crude product) as a white solid.

ESI-MS m/z:469.00[M+H]+。ESI-MS m/z: 469.00 [M+H]+ .

步骤2：化合物2-2的合成Step 2: Synthesis of compound 2-2

在室温下，将甲醇钠(224.56mg)加入到2-1(390mg)的甲苯(15mL)中，移至110℃回流搅拌5小时。待反应完全后，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机相经Na2SO4干燥且真空浓缩，得到白色固体状粗品目标产物，即化合物2-2(350mg，粗品)。Sodium methoxide (224.56 mg) was added to toluene (15 mL) of 2-1 (390 mg) at room temperature, and the mixture was moved to 110° C. under reflux and stirred for 5 hours. After the reaction was complete, the mixture was poured into ice water, extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo to obtain the crude target product, compound 2-2 (350 mg, crude) as a white solid.

ESI-MS m/z:450.94[M+H]+。ESI-MS m/z: 450.94 [M+H]+ .

步骤3：化合物2-3的合成Step 3: Synthesis of Compound 2-3

在室温下，氮气保护下，将DIEA(1.0mL)加入到化合物2-2(350mg)的POCl3(10.0mL) 中，移至110℃搅拌12小时。待反应完全后，直接浓缩除去POCl3，得到棕褐色固体状粗品目标产物，即化合物2-3(340mg，粗品)。At room temperature, under nitrogen protection, DIEA (1.0 mL) was added to POCl3 (10.0 mL) of compound 2-2 (350 mg), moved to 110° C. and stirred for 12 hours. After the reaction was complete, the reaction was directly concentrated to remove POCl3 to obtain the crude target product, compound 2-3 (340 mg, crude product) as a tan solid.

ESI-MS m/z:468.87[M+H]+。ESI-MS m/z: 468.87 [M+H]+.

步骤4：化合物2-4的合成Step 4: Synthesis of compounds 2-4

在室温下，将DIEA(279mg)加入到化合物2-3(340mg)，2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(163mg)的1,4-二氧六环(10mL)中，室温搅拌5分钟。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(PE:EA＝10:1-15:1)纯化，得到黄色固体状的所要目标产物，即化合物2-4(348mg，42.62％产率)。At room temperature, DIEA (279 mg) was added to compound 2-3 (340 mg), 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (163 mg) in 1,4-dioxo Hexacyclone (10 mL), stirred at room temperature for 5 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (PE:EA=10:1-15:1) Purification afforded the desired target product, compound 2-4 (348 mg, 42.62% yield) as a yellow solid.

ESI-MS m/z:659.06[M+H]+。ESI-MS m/z: 659.06 [M+H]+ .

步骤5：化合物2-5的合成Step 5: Synthesis of Compound 2-5

在氮气保护下，将三氟乙醇(158.4mg)加入到化合物2-4(348mg)，Cs2CO3(343.92mg，)的1,4-二氧六环(5mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物，即化合物2-5(400mg，粗品)。Under nitrogen protection, trifluoroethanol (158.4 mg) was added to compound 2-4 (348 mg), Cs2 CO3 (343.92 mg,) in 1,4-dioxane (5 mL), and moved to 100°C React for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product, compound 2-5 (400 mg, crude) as a yellow solid.

ESI-MS m/z:739.01[M+H]+。ESI-MS m/z: 739.01 [M+H]+ .

步骤6：化合物2-6的合成Step 6: Synthesis of Compound 2-6

在氮气保护下，将Pd(dppf)Cl2.CH2Cl2(63.82mg)加入到化合物2-5(400mg)，K2CO3(149.53mg)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(83.32mg，)的1,4-二氧六环(4.0mL)，H2O(0.4mL)混合溶液中，在70℃搅拌1h。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(PE:EA＝10:1-15:1)纯化，得到黄色固体状的所要目标产物，即化合物2-6(213mg，61.56％产率)。Under nitrogen protection, Pd(dppf)Cl2 .CH2 Cl2 (63.82 mg) was added to compound 2-5 (400 mg), K2 CO3 (149.53 mg), 4,4,5,5-tetramethyl In a mixed solution of 2-vinyl-1,3,2-dioxaborolane (83.32mg,) in 1,4-dioxane (4.0mL), H2 O (0.4mL) , stirred at 70 °C for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography ( PE:EA=10:1-15:1) to obtain the desired target product, compound 2-6 (213 mg, 61.56% yield) as a yellow solid.

ESI-MS m/z:639.01[M+H]+。ESI-MS m/z: 639.01 [M+H]+ .

步骤7：化合物2-7的合成Step 7: Synthesis of Compound 2-7

在氮气保护下，将Pd(PPh3)4(116.16mg)加入到化合物2-6(213mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(227.96mg)，K3PO4(141.39mg，)的1,4-二氧六环(2mL),H2O(0.5mL)溶液中，移至85℃反应反应约3小时。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥 且真空浓缩，浓缩物通过pre-TLC(PE:EA＝2:1)纯化,得到呈黄色固体状的所要产物，即化合物2-7(180mg，69.74％产率)。ESI-MS m/z:775.42[M+H]+。Under nitrogen protection, Pd(PPh3 )4 (116.16mg) was added to compound 2-6 (213mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (227.96mg), K3 PO4 (141.39mg,) in 1,4-dioxane (2mL ), H2 O (0.5 mL) solution, moved to 85°C for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) to obtain the desired product, compound 2-7 (180 mg, 69.74% yield) as a yellow solid. ESI-MS m/z: 775.42 [M+H]+ .

步骤8：化合物2-8的合成Step 8: Synthesis of Compound 2-8

在室温下，将化合物2-7(180mg)溶于DCM(6mL)和TFA(3mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色固体状粗品目标产物，即化合物2-8(135mg，粗品)。Compound 2-7 (180 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product, Compound 2-8 (135 mg, crude product) as a yellow solid.

ESI-MS m/z:591.40[M+H]+。ESI-MS m/z: 591.40 [M+H]+ .

[根据细则91更正 15.02.2022]步骤9：化合物2的合成[Corrected under Rule 91 15.02.2022]Step 9: Synthesis ofCompound 2

在氮气保护下，冰水浴中，将丙烯酰氯(17.03mg)的THF溶液加入到化合物2-8(135mg，)，THF(4mL)，饱和Na2CO3(1mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物，即化合物2(66.71mg，44.36％产率)。ESI-MS m/z:645.44[M+H]+。1HNMR(500MHz,CDCl3)δ：7.94(s,1H),7.45-7.48(m,2H),7.36-7.36(m,1H),6.37-6.41(m,1H),6.22-6.28(m,2H),5.70-5.73(m,1H),5.62-5.66(m,1H),5.05-5.07(m,1H),4.83-4.91(m,1H),4.47-4.54(m,1H),4.04(s,2H),3.94(s,2H),3.70-3.79(m,4H),2.81-2.87(m,1H),2.15(s,3H),2.07-2.09(m,2H),1.85-1.87(m,2H),1.74-1.77(m,1H),1.61-1.71(m,3H),1.41-1.48(m,3H),1.25-1.37(m,3H)。Under nitrogen protection, in an ice-water bath, a THF solution of acryloyl chloride (17.03 mg) was added to compound 2-8 (135 mg, ), THF (4 mL), saturated Na2 CO3 (1 mL) solution, and the reaction was complete upon addition . The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product, compound 2 (66.71 mg, 44.36% yield) as a yellow solid. ESI-MS m/z: 645.44 [M+H]+ . 1HNMR (500MHz, CDCl3) δ: 7.94(s,1H),7.45-7.48(m,2H),7.36-7.36(m,1H),6.37-6.41(m,1H),6.22-6.28(m,2H) ,5.70-5.73(m,1H),5.62-5.66(m,1H),5.05-5.07(m,1H),4.83-4.91(m,1H),4.47-4.54(m,1H),4.04(s, 2H),3.94(s,2H),3.70-3.79(m,4H),2.81-2.87(m,1H),2.15(s,3H),2.07-2.09(m,2H),1.85-1.87(m, 2H), 1.74-1.77(m, 1H), 1.61-1.71(m, 3H), 1.41-1.48(m, 3H), 1.25-1.37(m, 3H).

[根据细则91更正 15.02.2022]实施例3：化合物3(1-(7-(2-四氢呋喃-3-基-7-(5-甲基-1H-吲唑-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2，7-二氮杂螺[3.5]壬烷-2-基)丙-2-烯-1-酮)的合成[Corrected under Rule 91 15.02.2022]Example 3: Compound 3 (1-(7-(2-tetrahydrofuran-3-yl-7-(5-methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoro Synthesis of ethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]nonan-2-yl)prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000042
Figure PCTCN2021124884-APPB-000042

步骤1：化合物3-1的合成Step 1: Synthesis of compound 3-1

在室温下，将四氢呋喃-3-甲酰氯(402mg)加入到化合物M1(300mg)的THF(10mL)中，移至40℃搅拌4小时。待反应完全后，直接浓缩出去THF，得到黄色固体状粗品目标产物，即化合物3-1(460mg，粗品)。Tetrahydrofuran-3-carbonyl chloride (402 mg) was added to compound M1 (300 mg) in THF (10 mL) at room temperature, and the mixture was moved to 40°C and stirred for 4 hours. After the reaction was complete, THF was directly concentrated to obtain the crude target product as a yellow solid, namely compound 3-1 (460 mg, crude product).

ESI-MS m/z:456.94[M+H]+。ESI-MS m/z: 456.94 [M+H]+ .

步骤2：化合物3-2的合成Step 2: Synthesis of compound 3-2

在室温下，将甲醇钠(224.56mg)加入到化合物3-1(460mg，0.83mmol)的甲苯(10mL)中，移至110℃回流搅拌8小时。待反应完全后，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机相经Na2SO4干燥且真空浓缩，得到白色固体状粗品目标产物，即化合物3-2(200mg，粗品)。Sodium methoxide (224.56 mg) was added to compound 3-1 (460 mg, 0.83 mmol) in toluene (10 mL) at room temperature, moved to 110° C. under reflux and stirred for 8 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo to obtain the crude target product, compound 3-2 (200 mg, crude) as a white solid.

ESI-MS m/z:438.96[M+H]+。ESI-MS m/z: 438.96 [M+H]+.

步骤3：化合物3-3的合成Step 3: Synthesis of compound 3-3

在室温下，氮气保护下，将DIEA(1.0mL)加入到化合物3-2(200mg)的POCl3(10.0mL)中，移至110℃搅拌12小时。待反应完全后，直接浓缩除去POCl3，得到棕褐色固体状粗品目标产物，即化合物3-3(230mg，粗品)。At room temperature, under nitrogen protection, DIEA (1.0 mL) was added into POCl3 (10.0 mL) of compound 3-2 (200 mg), moved to 110° C. and stirred for 12 hours. After the reaction was complete, the reaction was directly concentrated to remove POCl3 to obtain the crude target product, Compound 3-3 (230 mg, crude product) as a tan solid.

ESI-MS m/z:456.87[M+H]+。ESI-MS m/z: 456.87 [M+H]+ .

步骤4：化合物3-4的合成Step 4: Synthesis of Compound 3-4

在室温下，将DIEA(279mg)加入到化合物3-3(200mg)，2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(90mg)的1,4-二氧六环(2mL)中，室温搅拌5分钟。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过爬大板纯化，得到黄色固体状的所要目标产物，即化合物3-4(88mg，34.1％产率)。At room temperature, DIEA (279 mg) was added to compound 3-3 (200 mg), 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (90 mg) in 1,4-dioxo Hexacyclic (2 mL), stirred at room temperature for 5 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried overNa2SO4 and concentrated invacuo , the residue was purified by climbing plates to give the desired target product, compound 3, as a yellow solid -4 (88 mg, 34.1% yield).

ESI-MS m/z:647.14[M+H]+。ESI-MS m/z: 647.14 [M+H]+ .

步骤5：化合物3-5的合成Step 5: Synthesis of Compound 3-5

在氮气保护下，将三氟乙醇(68mg)加入到化合物3-4(88mg)，Cs2CO3(88mg)的1,4-二氧六环(2mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物，即化合物3-5(97mg，粗品)。Under nitrogen protection, trifluoroethanol (68 mg) was added to compound 3-4 (88 mg), Cs2 CO3 (88 mg) in 1,4-dioxane (2 mL), and moved to 100° C. for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product, compound 3-5 (97 mg, crude) as a yellow solid.

ESI-MS m/z:727.08[M+H]+。ESI-MS m/z: 727.08 [M+H]+ .

步骤6：化合物3-6的合成Step 6: Synthesis of Compound 3-6

在氮气保护下，将Pd(dppf)Cl2.CH2Cl2(10mg)加入到化合物3-5(90mg)，K2CO3(34mg，)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(19mg)的1,4-二氧六环(2.5mL)，H2O(0.5mL)混合溶液中，在70℃搅拌1h。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(PE:EA＝10:1-3:1)纯化，得到黄色固体状的所要目标产物，即化合物3-6(213mg，64.4％产率)。Under nitrogen protection, Pd(dppf)Cl2 .CH2 Cl2 (10 mg) was added to compound 3-5 (90 mg), K2 CO3 (34 mg,), 4,4,5,5-tetramethyl -2-vinyl-1,3,2-dioxaborolane (19mg) in 1,4-dioxane (2.5mL), H2 O (0.5mL) mixed solution at 70 Stir at ℃ for 1h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography ( PE:EA=10:1-3:1) to obtain the desired target product, compound 3-6 (213 mg, 64.4% yield) as a yellow solid.

ESI-MS m/z:627.21[M+H]+。ESI-MS m/z: 627.21 [M+H]+ .

步骤7：化合物3-7的合成Step 7: Synthesis of Compound 3-7

在氮气保护下，将Pd(PPh3)4(18.5mg)加入到化合物3-6(50mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(54.6mg)，K3PO4(34mg)的1,4-二氧六环(2mL),H2O(0.5mL)溶液中，移至85℃反应反应约4小时。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(PE:EA＝2:1)纯化,得到呈黄色固体状的所要产物，即化合物3-7(45mg，73.8％产率)。Under nitrogen protection, Pd(PPh3 )4 (18.5 mg) was added to compound 3-6 (50 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 , 5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (54.6 mg), K3 PO4 (34 mg) in 1,4-dioxane (2 mL), H2 O (0.5 mL) solution, moved to 85°C for about 4 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) to obtain the desired product, compound 3-7 (45 mg, 73.8% yield) as a yellow solid.

ESI-MS m/z:763.39[M+H]+。ESI-MS m/z: 763.39 [M+H]+ .

步骤8：化合物3-8的合成Step 8: Synthesis of Compound 3-8

在室温下，将化合物3-7(45mg)溶于DCM(1mL)和TFA(0.5mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色固体状粗品目标产物，即化合物3-8(56mg，粗品)。ESI-MS m/z:579.32[M+H]+。Compound 3-7 (45 mg) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product, Compound 3-8 (56 mg, crude product) as a yellow solid. ESI-MS m/z: 579.32 [M+H]+ .

[根据细则91更正 15.02.2022]步骤9：化合物3的合成[Corrected under Rule 91 15.02.2022]Step 9: Synthesis of compound 3

在氮气保护下，冰水浴中，将丙烯酰氯(5.85mg)的THF溶液加入到化合物3-8(56mg，)，THF(2mL)，饱和Na2CO3(0.5mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化，得到呈黄色固体状的所要产物，即化合物3(12.05mg，32.2％产率)。ESI-MS m/z:633.41[M+H]+。Under the protection of nitrogen, in an ice-water bath, the THF solution of acryloyl chloride (5.85mg) was added to the compound 3-8 (56mg,), THF (2mL), saturated Na2 CO3 (0.5mL) solution, added to react completely. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product, compound 3 (12.05 mg, 32.2% yield) as a yellow solid. ESI-MS m/z: 633.41 [M+H]+.

1H NMR(500MHz,Chloroform-d)δ7.88(s,1H),7.40(d,J＝7.5Hz,2H),7.19(s,1H),6.32(dd,J＝17.0,1.9Hz,1H),6.18(m,J＝17.0,10.6,4.9Hz,2H),5.65(dd,J＝10.3,1.9Hz,1H),5.58(dd,J＝17.4,1.0Hz,1H),5.01(dd,J＝10.9,1.0Hz,1H),4.78–4.64(m,1H),4.34(m,1H),4.20(td,J＝8.1,3.7Hz,1H),4.05(dd,J＝8.3,6.8Hz,1H),4.02–3.95(m,3H),3.94–3.89(m,1H),3.87(s,2H),3.75(td,J＝10.0,9.0,5.5Hz,2H),3.71–3.62(m,3H),2.41(m,1H),2.29(m,1H),2.08(s,3H),1.18(d,J＝7.2Hz,4H)。1H NMR (500MHz, Chloroform-d) δ7.88(s,1H),7.40(d,J=7.5Hz,2H),7.19(s,1H),6.32(dd,J=17.0,1.9Hz,1H) ,6.18(m,J=17.0,10.6,4.9Hz,2H),5.65(dd,J=10.3,1.9Hz,1H),5.58(dd,J=17.4,1.0Hz,1H),5.01(dd,J =10.9,1.0Hz,1H),4.78–4.64(m,1H),4.34(m,1H),4.20(td,J=8.1,3.7Hz,1H),4.05(dd,J=8.3,6.8Hz, 1H),4.02–3.95(m,3H),3.94–3.89(m,1H),3.87(s,2H),3.75(td,J＝10.0,9.0,5.5Hz,2H),3.71–3.62(m, 3H), 2.41 (m, 1H), 2.29 (m, 1H), 2.08 (s, 3H), 1.18 (d, J=7.2Hz, 4H).

实施例4：化合物4(1-(7-(6-乙基-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-8-(2,2,2-三氟乙氧基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 4: Compound 4 (1-(7-(6-ethyl-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl) -8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-ene-1- ketone) synthesis

Figure PCTCN2021124884-APPB-000043
Figure PCTCN2021124884-APPB-000043

步骤1：化合物4-1的合成Step 1: Synthesis of Compound 4-1

在室温下，在氢气氛围中，将Pd/C(80mg)加入到化合物1-9(95mg)的MeOH(10mL)的溶液中，在室温搅拌2小时。待反应完全后，过滤除去Pd/C，并用MeOH洗涤Pd/C，将滤液浓缩，即得黄色固体粗品，即化合物4-1(70mg，粗品)。Pd/C (80 mg) was added to a solution of compound 1-9 (95 mg) in MeOH (10 mL) under a hydrogen atmosphere at room temperature, and stirred at room temperature for 2 hours. After the reaction was complete, the Pd/C was removed by filtration, and the Pd/C was washed with MeOH, and the filtrate was concentrated to obtain a crude yellow solid, namely compound 4-1 (70 mg, crude).

ESI-MS m/z:608.43[M+H]+。ESI-MS m/z: 608.43 [M+H]+ .

步骤2：化合物4的合成Step 2: Synthesis of compound 4

在氮气保护下，冰水浴中，将丙烯酰氯(12.51mg)的THF溶液加入到化合物4-1(70mg)，THF(4mL)，饱和Na2CO3(1mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物，即化合物4(18.3mg，23.79％产率)。Under nitrogen protection, a THF solution of acryloyl chloride (12.51 mg) was added to compound 4-1 (70 mg), THF (4 mL), saturated Na2 CO3 (1 mL) solution in an ice-water bath, and the reaction was complete upon addition. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product, compound 4 (18.3 mg, 23.79% yield) as a yellow solid.

ESI-MS m/z:662.37[M+H]+。ESI-MS m/z: 662.37 [M+H]+ .

实施例5：化合物5(1-(7-(7-(5-甲基-1H-吲唑-4-基)-2-(四氢-2H-吡喃-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Embodiment 5: Compound 5 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(tetrahydro-2H-pyran-4-yl)-8-( 2,2,2-Trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-ene-1 - ketone) synthesis

Figure PCTCN2021124884-APPB-000044
Figure PCTCN2021124884-APPB-000044

步骤1：化合物5-1的合成Step 1: Synthesis of Compound 5-1

在室温下，将四氢吡喃-4-甲酰氯(248.38mg，1.67mmol)加入到化合物M1(300mg，)的THF(3mL)中，移至40℃搅拌4小时。待反应完全后，直接浓缩出去THF，得到白色固体状粗品目标产物，即化合物5-1(390mg，粗品)。Tetrahydropyran-4-carbonyl chloride (248.38 mg, 1.67 mmol) was added to compound M1 (300 mg, ) in THF (3 mL) at room temperature, moved to 40° C. and stirred for 4 hours. After the reaction was complete, THF was directly concentrated to obtain the crude target product as a white solid, namely compound 5-1 (390 mg, crude product).

ESI-MS m/z:470.92[M+H]+。ESI-MS m/z: 470.92 [M+H]+ .

步骤2：化合物5-2的合成Step 2: Synthesis of compound 5-2

在室温下，将甲醇钠(224.56mg)加入到化合物5-1(390mg)的甲苯(4mL)中，移至110℃回流搅拌5小时。待反应完全后，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机相经Na2SO4干燥且真空浓缩，得到白色固体状粗品目标产物，即化合物5-2(350mg，粗品)。Sodium methoxide (224.56 mg) was added to compound 5-1 (390 mg) in toluene (4 mL) at room temperature, and the mixture was moved to 110° C. under reflux and stirred for 5 hours. After the reaction was complete, the mixture was poured into ice water, extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo to obtain the crude target product, compound 5-2 (350 mg, crude) as a white solid.

ESI-MS m/z:452.96[M+H]+。ESI-MS m/z: 452.96 [M+H]+ .

步骤3：化合物5-3的合成Step 3: Synthesis of compound 5-3

在室温下，氮气保护下，将DIEA(0.4mL)加入到化合物5-2(350mg,0.77mmol)的POCl3(4mL)中，移至110℃搅拌12小时。待反应完全后，直接浓缩除去POCl3，得到棕褐色固体状粗品目标产物，即化合物5-3(210mg，粗品)。At room temperature, under nitrogen protection, DIEA (0.4 mL) was added to compound 5-2 (350 mg, 0.77 mmol) in POCl3 (4 mL), moved to 110° C. and stirred for 12 hours. After the reaction was complete, the reaction was directly concentrated to remove POCl3 to obtain the crude target product, compound 5-3 (210 mg, crude product) as a tan solid.

ESI-MS m/z:470.95[M+H]+。ESI-MS m/z: 470.95 [M+H]+ .

步骤4：化合物5-4的合成Step 4: Synthesis of compound 5-4

在室温下，将DIEA(115.13mg)加入到化合物5-3(210mg)，2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(120.96mg)的1,4-二氧六环(3mL)中，室温搅拌30分钟。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(PE:EA＝10:1-15:1)纯化，得到黄色固体状的所要目标产物，即化合物5-4(110mg，37.34％产率)。At room temperature, DIEA (115.13mg) was added to compound 5-3 (210mg), 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (120.96mg) in 1,4- Dioxane (3 mL), stirred at room temperature for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (PE:EA=10:1-15:1) Purification afforded the desired target product, compound 5-4 (110 mg, 37.34% yield) as a yellow solid.

ESI-MS m/z:661.17[M+H]+。ESI-MS m/z: 661.17 [M+H]+ .

步骤5：化合物5-5的合成Step 5: Synthesis of Compound 5-5

在氮气保护下，将三氟乙醇(49.92mg)加入到化合物5-4(110mg)，Cs2CO3(108.39mg，)的1,4-二氧六环(2mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物，即化合物5-5(105mg，粗品)。Under nitrogen protection, trifluoroethanol (49.92mg) was added to compound 5-4 (110mg), Cs2 CO3 (108.39mg,) in 1,4-dioxane (2mL), and moved to 100°C React for 30 minutes. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product, compound 5-5 (105 mg, crude) as a yellow solid.

ESI-MS m/z:741.14[M+H]+。ESI-MS m/z: 741.14 [M+H]+ .

步骤6：化合物5-6的合成Step 6: Synthesis of Compound 5-6

在氮气保护下，将Pd(dppf)Cl2.CH2Cl2(11.56mg)加入到化合物5-5(105mg)，K2CO3(39.15mg)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(21.81mg)的1,4-二氧六环(2mL)，H2O(0.4mL)混合溶液中，在70℃搅拌1h。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓 缩，剩余物通过快速硅胶柱色谱(PE:EA＝10:1-15:1)纯化，得到黄色固体状的所要目标产物，即化合物5-6(60mg，66.04％产率)。ESI-MS m/z:641.25[M+H]+。Under nitrogen protection, Pd(dppf)Cl2 .CH2 Cl2 (11.56 mg) was added to compound 5-5 (105 mg), K2 CO3 (39.15 mg), 4,4,5,5-tetramethyl In a mixed solution of 1,4-dioxane (2 mL) and H2 O (0.4 mL) of 2-ethenyl-2-vinyl-1,3,2-dioxaborolane (21.81 mg), in Stir at 70°C for 1h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was passed through flash silica gel column chromatography ( PE:EA=10:1-15:1) to obtain the desired target product, compound 5-6 (60 mg, 66.04% yield) as a yellow solid. ESI-MS m/z: 641.25 [M+H]+ .

步骤7：化合物5-7的合成Step 7: Synthesis of Compound 5-7

在氮气保护下，将Pd(PPh3)4(21.60mg)加入到化合物5-6(60mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(64.02mg)，K3PO4(39.71mg，)的1,4-二氧六环(2mL),H2O(0.2mL)溶液中，移至85℃反应反应约3小时。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(PE:EA＝2:1)纯化,得到呈黄色固体状的所要产物，即化合物5-7(25mg，34.41％产率)。Under nitrogen protection, Pd(PPh3 )4 (21.60 mg) was added to compound 5-6 (60 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (64.02mg), K3 PO4 (39.71mg,) in 1,4-dioxane (2mL ), H2 O (0.2 mL) solution, moved to 85°C for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) to obtain the desired product, compound 5-7 (25 mg, 34.41% yield) as a yellow solid.

ESI-MS m/z:777.42[M+H]+。ESI-MS m/z: 777.42 [M+H]+ .

步骤8：化合物5-8的合成Step 8: Synthesis of Compound 5-8

在室温下，将化合物5-7(25mg)溶于DCM(1mL)和TFA(0.5mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色油状粗品目标产物，即化合物5-8(20mg，粗品)。Compound 5-7 (25 mg) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product in the form of yellow oil, namely compound 5-8 (20 mg, crude product).

ESI-MS m/z:593.33[M+H]+。ESI-MS m/z: 593.33 [M+H]+ .

步骤9：化合物5的合成Step 9: Synthesis of Compound 5

在氮气保护下，冰水浴中，将丙烯酰氯(3.20mg)的THF溶液加入到化合物5-8(20mg，)，THF(2mL)，饱和Na2CO3(0.5mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化，得到呈黄色固体状的所要产物，即化合物5(5.2mg，33.48％产率)。Under the protection of nitrogen, in an ice-water bath, a THF solution of acryloyl chloride (3.20mg) was added to compound 5-8 (20mg,), THF (2mL), saturated Na2 CO3 (0.5mL) solution, added to react completely. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product, compound 5 (5.2 mg, 33.48% yield) as a yellow solid.

ESI-MS m/z:647.33[M+H]+。ESI-MS m/z: 647.33 [M+H]+ .

实施例6：化合物6(1-(7-(7-(5-甲基-1H-吲唑-4-基)-2-(N-甲基吡咯-3-基)-8-(2，2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Embodiment 6: Compound 6 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(N-methylpyrrol-3-yl)-8-(2, 2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one )Synthesis

Figure PCTCN2021124884-APPB-000045
Figure PCTCN2021124884-APPB-000045

步骤1：化合物6-1的合成Step 1: Synthesis of compound 6-1

在室温、氮气保护下，将N-甲基吡咯-3-羧酸(600mg)溶于SOCl2(5.5mL)中，升温至70℃，搅拌反应1h。真空浓缩，除去氯化亚砜，得到白色固体目标产物，即化合物6-1(68mg，粗品)。At room temperature under nitrogen protection, N-methylpyrrole-3-carboxylic acid (600 mg) was dissolved in SOCl2 (5.5 mL), heated to 70° C., and stirred for 1 h. Concentrate in vacuo to remove thionyl chloride to obtain the target product, compound 6-1 (68 mg, crude product) as a white solid.

步骤2：化合物6-2的合成Step 2: Synthesis of Compound 6-2

在室温下，将化合物6-1(369mg)加入到化合物M1(300mg)的THF(5mL)中，搅拌16小时。待反应完全后，旋蒸除去THF。使用二氯甲烷：甲醇＝90:10作为展开剂，柱层析分离得到淡黄色固体目标产物，即化合物6-2(262mg)。Compound 6-1 (369 mg) was added to compound M1 (300 mg) in THF (5 mL) at room temperature, and stirred for 16 hours. After the reaction was complete, THF was removed by rotary evaporation. Using dichloromethane:methanol=90:10 as a developing solvent, the target product, compound 6-2 (262 mg), was obtained through column chromatography separation.

ESI-MS m/z:469.93,471.94[M+H]+。ESI-MS m/z: 469.93, 471.94 [M+H]+ .

步骤3：化合物6-3的合成Step 3: Synthesis of Compound 6-3

在室温下，将甲醇钠(151mg)加入到化合物6-2(262mg)的甲苯(10mL)中，升温至110℃回流搅拌16小时。待反应完全后，将混合物倾入40mL冰水中，用乙酸乙酯萃取该混合物，有机相经Na2SO4干燥且真空浓缩，得到淡黄色固体目标产物，即化合物6-3(240mg，粗品)。Sodium methoxide (151 mg) was added to compound 6-2 (262 mg) in toluene (10 mL) at room temperature, and the mixture was heated to 110° C. under reflux and stirred for 16 hours. After the reaction was complete, the mixture was poured into 40 mL of ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo to obtain the target product as a pale yellow solid, compound 6-3 (240 mg, crude product) .

ESI-MS m/z:451.96,453.91[M+H]+。ESI-MS m/z: 451.96, 453.91 [M+H]+ .

步骤4：化合物6-4的合成Step 4: Synthesis of compound 6-4

在室温、氮气保护下，将DIEA(0.5mL)加入到化合物6-3(226mg)的POCl3(5mL)溶液中，升温至110℃搅拌3小时。待反应完全后，旋蒸除去POCl3，将混合物倒入冰水中，用乙酸乙酯萃取混合物，收集有机相，用饱和卤水洗涤，经Na2SO4干燥且真空浓缩，得到棕黄色固体目标产物，即化合物6-4(116mg，粗品)。At room temperature under nitrogen protection, DIEA (0.5 mL) was added to a solution of compound 6-3 (226 mg) in POCl3 (5 mL), heated to 110° C. and stirred for 3 hours. After the reaction was complete, POCl3 was removed by rotary evaporation, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was collected, washed with saturated brine, dried over Na2 SO4 and concentrated in vacuo to obtain the target product as a brown yellow solid , namely compound 6-4 (116 mg, crude product).

ESI-MS m/z:469.96,471.94[M+H]+。ESI-MS m/z: 469.96, 471.94 [M+H]+ .

步骤5：化合物6-5的合成Step 5: Synthesis of compound 6-5

在室温下，将DIEA(96mg)加入到化合物6-4(116mg)，2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(56mg)的二氧六环(5mL)溶液中，室温搅拌2小时。反应结束后，旋干二氧六环，加入冰水，用乙酸乙酯/甲醇＝10:1萃取混合物，收集有机相旋干。剩余物通过硅胶柱层析色谱(展开剂为DCM:MeOH＝10:1)纯化，得到黄色固体目标产物，即化合物6-5(58mg)。DIEA (96 mg) was added to compound 6-4 (116 mg), tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (56 mg) in dioxane (5 mL) at room temperature ) solution, stirred at room temperature for 2 hours. After the reaction, the dioxane was spin-dried, ice water was added, the mixture was extracted with ethyl acetate/methanol=10:1, and the organic phase was collected and spin-dried. The residue was purified by silica gel column chromatography (developing solvent: DCM:MeOH=10:1) to obtain the target product, compound 6-5 (58 mg), as a yellow solid.

ESI-MS m/z:660.08,662.07[M+H]+。ESI-MS m/z: 660.08, 662.07 [M+H]+ .

步骤6：化合物6-6的合成Step 6: Synthesis of compound 6-6

向充分干燥的反应瓶中加入化合物6-5(56mg)，用二氧六环(3mL)溶解，随后加入三氟乙醇(42mg)及Cs2CO3(55mg)，升温至100℃反应1h。反应结束后，用THF稀释反应液，过滤除去Cs2CO3。滤液旋干后，使用隔膜泵干燥样品，得到黄色固体目标产物，即化合物6-6(60mg)。Compound 6-5 (56 mg) was added to a well-dried reaction flask, dissolved in dioxane (3 mL), then trifluoroethanol (42 mg) and Cs2 CO3 (55 mg) were added, and the temperature was raised to 100° C. for 1 h. After the reaction, the reaction solution was diluted with THF, and Cs2 CO3 was removed by filtration. After the filtrate was spin-dried, the sample was dried using a diaphragm pump to obtain the target product, Compound 6-6 (60 mg), as a yellow solid.

ESI-MS m/z:740.03,742.07[M+H]+。ESI-MS m/z: 740.03, 742.07 [M+H]+ .

步骤7：化合物6-7的合成Step 7: Synthesis of Compound 6-7

N2保护下，向干燥反应瓶中加入化合物6-6(60mg)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(13mg)，用二氧六环(5mL)溶解，随后加入K2CO3(22.40mg)、H2O(0.5mL)，分散均匀后，加入1,1'-双二苯基膦二茂铁二氯化钯(5.93mg)，N2置换体系三次。升温至70℃搅拌1h。反应结束后，将反应液冷却至室温，加入水后用乙酸乙酯:甲醇＝10:1萃取，饱和食盐水反萃取，收集有机相，经Na2SO4干燥、过滤、旋干，浓缩物通过pre-TLC(展开剂为DCM/MeOH＝10:1)纯化,得到黄色固体产物，即化合物6-7(44mg)。Under the protection ofN2 , compound 6-6 (60 mg), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane was added to the dry reaction flask (13mg), dissolved with dioxane (5mL), then added K2 CO3 (22.40mg), H2 O (0.5mL), after uniform dispersion, added 1,1'-bisdiphenylphosphine Iron palladium dichloride (5.93mg), N2 replacement system three times. Raise the temperature to 70°C and stir for 1h. After the reaction, the reaction liquid was cooled to room temperature, added water, extracted with ethyl acetate:methanol=10:1, back-extracted with saturated brine, collected the organic phase, dried over Na2 SO4 , filtered, and spin-dried, the concentrate Purification by pre-TLC (developing solvent: DCM/MeOH=10:1) gave a yellow solid product, compound 6-7 (44 mg).

ESI-MS m/z:640.11,642.13[M+H]+。ESI-MS m/z: 640.11, 642.13 [M+H]+ .

步骤8：化合物6-8的合成Step 8: Synthesis of compounds 6-8

向氮气保护的反应瓶中加入化合物6-7(44mg)，用二氧六环(5mL)溶解后，依次加入5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(47mg)、 K3PO4(29mg)、去离子水(0.5mL)以及Pd(PPh3)4(8mg)。升温至85℃反应3小时。将反应混合物冷却至室温，加入水后用乙酸乙酯:甲醇＝10:1萃取，饱和食盐水反萃取，收集有机相，经Na2SO4干燥、过滤、旋干，浓缩物通过pre-TLC(展开剂为DCM/MeOH＝10:1)纯化，得到黄色固体产物，即化合物6-8(18mg)。ESI-MS m/z:776.87[M+H]+。Add compound 6-7 (44mg) to the nitrogen-protected reaction flask, after dissolving with dioxane (5mL), add 5-methyl-1-tetrahydropyran-2-yl-4-(4, 4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (47mg), K3 PO4 (29mg), deionized water (0.5mL) and Pd( PPh3 )4 (8 mg). Raise the temperature to 85°C for 3 hours. The reaction mixture was cooled to room temperature, added water, extracted with ethyl acetate:methanol=10:1, back-extracted with saturated brine, collected the organic phase, dried over Na2 SO4 , filtered, and spin-dried, and the concentrate was passed through pre-TLC Purification (developing solvent: DCM/MeOH=10:1) gave a yellow solid product, compound 6-8 (18 mg). ESI-MS m/z: 776.87 [M+H]+ .

步骤9：化合物6-9的合成Step 9: Synthesis of compounds 6-9

在室温下，将化合物6-8(18mg)溶于DCM(2mL)和TFA(1mL)的混合溶剂中，升温至40℃反应1h。待反应完全后，浓缩除去溶剂，得到黄色固体粗品产物，即化合物6-9(21mg，粗品)直接用于下一步投料。ESI-MS m/z:592.94[M+H]+。Compound 6-8 (18 mg) was dissolved in a mixed solvent of DCM (2 mL) and TFA (1 mL) at room temperature, and the temperature was raised to 40° C. for 1 h. After the reaction was complete, the solvent was concentrated and removed to obtain a yellow solid crude product, namely compound 6-9 (21 mg, crude product), which was directly used for feeding in the next step. ESI-MS m/z: 592.94 [M+H]+ .

步骤10：化合物6的合成Step 10: Synthesis of Compound 6

在氮气保护、冰水浴下，将丙烯酰氯(2.0mg)的THF(1mL)溶液滴加到化合物6-9(13mg，)的THF(4mL)与饱和NaHCO3(1mL)混合溶液中，搅拌10分钟。将反应液倒入水中，用乙酸乙酯:甲醇＝10:1萃取，饱和食盐水反萃取，收集有机相，经Na2SO4干燥、过滤、旋干，浓缩物通过pre-TLC(展开剂为DCM/MeOH＝10:1)纯化，得到淡黄色固体产物，即化合物6(7mg，LC-MS纯度96.2％)。Under nitrogen protection and ice-water bath, a solution of acryloyl chloride (2.0 mg) in THF (1 mL) was added dropwise to a mixed solution of compound 6-9 (13 mg,) in THF (4 mL) and saturated NaHCO3 (1 mL), and stirred for 10 minute. The reaction solution was poured into water, extracted with ethyl acetate:methanol= 10:1, back-extracted with saturated brine, the organic phase was collected, dried overNa2SO4 , filtered, and spin-dried, and the concentrate was passed through pre-TLC (developing solvent Purified (DCM/MeOH=10:1) to obtain a pale yellow solid product, compound 6 (7 mg, LC-MS purity 96.2%).

ESI-MS m/z:646.34[M+H]+。ESI-MS m/z: 646.34 [M+H]+ .

实施例7：化合物7(1-(7-(2-(1-乙基哌啶-4-基)-7-(5-甲基-1H-吲唑-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,7-二氮螺环[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 7: Compound 7 (1-(7-(2-(1-ethylpiperidin-4-yl)-7-(5-methyl-1H-indazol-4-yl)-8-(2 ,2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1- ketone) synthesis

Figure PCTCN2021124884-APPB-000046
Figure PCTCN2021124884-APPB-000046

步骤1：化合物7-1的合成Step 1: Synthesis of compound 7-1

在室温下，将碘乙烷(3.27g)，K2CO3(2.89g)加入到甲基哌啶-4-羧酸甲酯(1.5g,)的EtOH(15mL)溶液中，移至85℃回流反应1h。真空浓缩，除去EtOH，得到白色固体状目标产物7-1(1.7g，粗品)。Ethyl iodide (3.27 g), K2 CO3 (2.89 g) was added to a solution of methyl piperidine-4-carboxylate (1.5 g,) in EtOH (15 mL) at room temperature and moved to 85 ℃ reflux reaction for 1h. Concentration in vacuo to remove EtOH afforded the desired product 7-1 (1.7 g, crude) as a white solid.

ESI-MS m/z:172.06[M+H]+。ESI-MS m/z: 172.06 [M+H]+ .

步骤2：化合物7-2的合成Step 2: Synthesis of compound 7-2

在室温下，将LiOH(447.57mg)加入到化合物7-1(1.7g)的MeOH(10mL)溶液中，移至50℃反应1小时。向其中加入与LiOH等当量的稀盐酸溶液，使目标产物游离，直接旋干，剩余物通过快速硅胶柱色谱(DCM:MeOH＝92:8-90:10)纯化，得到黄色固体状的所要目标产物7-2(700mg，44.85％产率)。LiOH (447.57 mg) was added to a MeOH (10 mL) solution of compound 7-1 (1.7 g) at room temperature, and the reaction was carried out at 50° C. for 1 hour. Dilute hydrochloric acid solution equivalent to LiOH was added thereto to free the target product, spin dry directly, and the residue was purified by flash silica gel column chromatography (DCM:MeOH=92:8-90:10) to obtain the desired target product as a yellow solid Product 7-2 (700 mg, 44.85% yield).

ESI-MS m/z:158.15[M+H]+。ESI-MS m/z: 158.15 [M+H]+ .

步骤3：化合物7-3的合成Step 3: Synthesis of compound 7-3

在室温下，氮气保护下，将化合物7-2(400mg)溶于SOCl2(3mL)中，移至70℃下，搅拌反应1h。真空浓缩，除去氯化亚砜，得到白色固体状目标产物7-3(450mg，粗品)。At room temperature, under nitrogen protection, compound 7-2 (400 mg) was dissolved in SOCl2 (3 mL), moved to 70° C., and stirred for 1 h. Concentration in vacuo to remove thionyl chloride gave the target product 7-3 (450 mg, crude product) as a white solid.

步骤4：化合物7-4的合成Step 4: Synthesis of compound 7-4

在室温下，将化合物7-3(293.63mg)加入到化合物M1(300mg)的THF(10mL)中，移至45℃搅拌12小时。反应有大量固体析出，抽滤，固体即是目标产物7-4(150mg，粗品)。Compound 7-3 (293.63 mg) was added to compound M1 (300 mg) in THF (10 mL) at room temperature, moved to 45°C and stirred for 12 hours. A large amount of solids were precipitated in the reaction, and the solids were the target product 7-4 (150 mg, crude product) after suction filtration.

ESI-MS m/z:498.00[M+H]+。ESI-MS m/z: 498.00 [M+H]+ .

步骤5：化合物7-5的合成Step 5: Synthesis of compound 7-5

在室温下，将甲醇钠(81.34mg)加入到化合物7-4(150mg)的toluene(5mL)中，移至110℃回流搅拌4小时。待反应完全后，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机相经Na2SO4干燥且真空浓缩，得到棕色固体状粗品目标产物7-5(150mg，粗品)。Sodium methoxide (81.34 mg) was added to toluene (5 mL) of compound 7-4 (150 mg) at room temperature, and the mixture was moved to 110° C. under reflux and stirred for 4 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo to obtain the crude target product 7-5 (150 mg, crude) as a brown solid.

ESI-MS m/z:480.00[M+H]+。ESI-MS m/z: 480.00 [M+H]+ .

步骤6：化合物7-6的合成Step 6: Synthesis of Compound 7-6

在室温下，氮气保护下，将DIEA(1.0mL)加入到化合物7-5(150mg)的POCl3(5mL)中，移至110℃搅拌3小时。待反应完全后，直接浓缩除去POCl3，加入EA稀释，用水萃取三次，合并有机相，干燥浓缩，得到白色固体状目标产物7-6(150mg，粗品)。At room temperature, under nitrogen protection, DIEA (1.0 mL) was added into POCl3 (5 mL) of compound 7-5 (150 mg), moved to 110° C. and stirred for 3 hours. After the reaction was complete, directly concentrate to remove POCl3 , add EA to dilute, extract with water three times, combine the organic phases, dry and concentrate to obtain the target product 7-6 (150 mg, crude product) as a white solid.

ESI-MS m/z:497.99[M+H]+。ESI-MS m/z: 497.99 [M+H]+ .

步骤7：化合物7-7的合成Step 7: Synthesis of Compound 7-7

在室温下，将DIEA(116.65mg)加入到化合物7-6(150mg)，2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(68.09mg)的二氧六环(5mL)中，移至45℃搅拌1小时。将混合物倾注入冰水中，用乙酸乙酯/甲醇＝10:1萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(DCM:MeOH＝10:1)纯化，得到黄色固体状的所要目标产物7-7(50mg,24.14％产率)。At room temperature, DIEA (116.65 mg) was added to compound 7-6 (150 mg), 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (68.09 mg) in dioxane (5 mL), moved to 45°C and stirred for 1 hour. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (DCM:MeOH=10: 1) Purification afforded the desired target product 7-7 (50 mg, 24.14% yield) as a yellow solid.

ESI-MS m/z:688.14[M+H]+。ESI-MS m/z: 688.14 [M+H]+ .

步骤8：化合物7-8的合成Step 8: Synthesis of compounds 7-8

在氮气保护下，将三氟乙醇(21.80mg)加入到化合物7-7(50mg)，Cs2CO3(47.33mg)的二氧六环(1mL)中，移至100℃反应3h。将混合物倾注入冰水中，用乙酸乙酯/甲醇＝10:1萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物7-7(55mg，粗品)。Under nitrogen protection, trifluoroethanol (21.80 mg) was added to compound 7-7 (50 mg), Cs2 CO3 (47.33 mg) in dioxane (1 mL), and moved to 100° C. for 3 h. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product 7-7 as a yellow solid (55 mg, Crude).

ESI-MS m/z:768.18[M+H]+。ESI-MS m/z: 768.18 [M+H]+ .

步骤9：化合物7-9的合成Step 9: Synthesis of compounds 7-9

在氮气保护下，将Pd(dppf)Cl2.CH2Cl2(5.84mg)加入到化合物7-8(55mg)，K2CO3(19.78mg)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(11.02mg)的二氧六环(1.0mL)，H2O(0.1mL)混合溶液中，在70℃搅拌1h。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化，得到呈黄色固体状的所要产物7-9(25mg, 37.39μmol,52.24％产率)。Under nitrogen protection, Pd(dppf)Cl2 .CH2 Cl2 (5.84 mg) was added to compound 7-8 (55 mg), K2 CO3 (19.78 mg), 4,4,5,5-tetramethyl Dioxane (1.0 mL), H2 O (0.1 mL) mixed solution of 2-vinyl-1,3,2-dioxaborolane (11.02 mg) was stirred at 70°C 1h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1) to obtain the desired product 7-9 (25 mg, 37.39 μmol, 52.24% yield) as a yellow solid.

ESI-MS m/z:668.18[M+H]+。ESI-MS m/z: 668.18 [M+H]+ .

步骤10：化合物7-10的合成Step 10: Synthesis of compounds 7-10

在氮气保护下，将Pd(PPh3)4(8.64mg)加入到化合物7-9(25mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(25.59mg)，K3PO4(15.87mg,)的二氧六环(1mL)，H2O(0.25mL)溶液中，移至85℃反应反应约3小时。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物7-10(20mg,66.53％产率)。ESI-MSUnder nitrogen protection, Pd(PPh3 )4 (8.64 mg) was added to compound 7-9 (25 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (25.59 mg), K3 PO4 (15.87 mg,) in dioxane (1 mL), H2 O (0.25mL) solution, moved to 85°C and reacted for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1) to obtain the desired product 7-10 (20 mg, 66.53% yield) as a yellow solid. ESI-MS

m/z:804.56[M+H]+。m/z:804.56[M+H]+ .

步骤11：化合物7-11的合成Step 11: Synthesis of Compound 7-11

在室温下，将化合物7-10(20mg)溶于DCM(1mL)和TFA(0.5mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色固体状目标产物7-11(18mg，粗品)。Compound 7-10 (20 mg) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the target product 7-11 (18 mg, crude product) as a yellow solid.

ESI-MS m/z:620.28[M+H]+。ESI-MS m/z: 620.28 [M+H]+ .

步骤12：化合物7的合成Step 12: Synthesis ofCompound 7

在氮气保护下，冰水浴中，将丙烯酰氯(3.15mg)的THF溶液加入到化合物9-11(18mg,)，THF(2mL)，饱和Na2CO3(0.5mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化，得到呈黄色固体状的所要产物7(10.7mg，52.69％产率)。Under the protection of nitrogen, in an ice-water bath, the THF solution of acryloyl chloride (3.15mg) was added to the compound 9-11 (18mg,), THF (2mL), saturated Na2 CO3 (0.5mL) solution, added to react completely. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product 7 (10.7 mg, 52.69% yield) as a yellow solid.

ESI-MS m/z:674.37[M+H]+。ESI-MS m/z: 674.37 [M+H]+ .

1H NMR(500MHz,DMSO-d6)δ13.05(s,1H),8.30(s,1H),8.01(s,1H),7.50(d,J＝8.5Hz,1H),7.40(s,1H),7.33(d,J＝8.5Hz,1H),6.35(m,1H),6.18–6.05(m,2H),5.82–5.66(m,2H),5.11(d,J＝11.1Hz,1H),4.97(m,1H),4.68(m,1H),3.23–3.05(m,4H),2.80(m,1H),2.25(m,2H),2.05(d,J＝6.7Hz,5H),1.95(m,6H),1.62–1.55(m,1H),1.32(m,1H),1.07(t,J＝7.2Hz,3H),0.94(t,J＝7.3Hz,2H).1 H NMR (500MHz,DMSO-d6 )δ13.05(s,1H),8.30(s,1H),8.01(s,1H),7.50(d,J=8.5Hz,1H),7.40(s, 1H), 7.33(d, J＝8.5Hz, 1H), 6.35(m, 1H), 6.18–6.05(m, 2H), 5.82–5.66(m, 2H), 5.11(d, J＝11.1Hz, 1H ),4.97(m,1H),4.68(m,1H),3.23–3.05(m,4H),2.80(m,1H),2.25(m,2H),2.05(d,J＝6.7Hz,5H) ,1.95(m,6H),1.62–1.55(m,1H),1.32(m,1H),1.07(t,J=7.2Hz,3H),0.94(t,J=7.3Hz,2H).

实施例8：化合物8(1-(7-(6-氯-8-乙氧基-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 8: Compound 8 (1-(7-(6-chloro-8-ethoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidine Synthesis of -4-yl)-quinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000047
Figure PCTCN2021124884-APPB-000047

步骤1：化合物8-1的合成Step 1: Synthesis of Compound 8-1

在氮气保护下，将乙醇(395mg)加入到化合物M2(1.0g)，Cs2CO3(1.68g)的二氧六环(10mL)中，移至80℃反应1h。降到室温，缓慢加入水，析出固体，过滤，烘干得到黄色固体8-1(0.96g，粗品)。Under nitrogen protection, ethanol (395 mg) was added to compound M2 (1.0 g), Cs2 CO3 (1.68 g) in dioxane (10 mL), and moved to 80° C. for 1 h. After cooling down to room temperature, water was slowly added to precipitate a solid, which was filtered and dried to obtain a yellow solid 8-1 (0.96 g, crude product).

ESI-MS m/z:608.19[M+H]+。ESI-MS m/z: 608.19 [M+H]+ .

步骤2：化合物8-2的合成Step 2: Synthesis of Compound 8-2

在氮气保护下，将Pd(PPh3)4(341mg)加入到化合物8-1(900mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(758.6mg)，K3PO4(941.1mg，)的二氧六环(20mL)，H2O(5mL)溶液中，移至85℃反应4h。降到室温，加水稀释，用EA萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，硅胶柱分离纯化(DCM:MeOH＝10:1)，得黄色固体8-2(665mg，49.9％产率)。Under nitrogen protection, Pd(PPh3 )4 (341 mg) was added to compound 8-1 (900 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (758.6 mg), K3 PO4 (941.1 mg,) in dioxane (20 mL), H2 O (5 mL) solution, moved to 85 ° C for 4 h. Cool down to room temperature, dilute with water, extract three times with EA, combine the organic phases and dry with anhydrous Na2 SO4 , filter, spin dry, and separate and purify on a silica gel column (DCM:MeOH=10:1) to obtain a yellow solid 8-2 (665 mg, 49.9% yield).

ESI-MS m/z:744.39[M+H]+。ESI-MS m/z: 744.39 [M+H]+ .

步骤3：化合物8-3的合成Step 3: Synthesis of compound 8-3

在室温条件下，将化合物8-2(120mg)溶于DCM(6mL)和TFA(3mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到棕色油粗品8-3(73mg，粗品)。Compound 8-2 (120 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain a brown oil crude product 8-3 (73 mg, crude product).

ESI-MS m/z:560.28[M+H]+。ESI-MS m/z: 560.28 [M+H]+ .

步骤4：化合物8的合成Step 4: Synthesis ofCompound 8

在氮气保护下，冰水浴中，将丙烯酰氯(12.1mg)的THF(1mL)溶液加入到化合物8-3(73mg)，THF(4mL)，饱和Na2CO3(1mL)溶液中，保持温度搅拌5min。加水稀释，用EA 萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，pre-TLC分离纯化(DCM:MeOH＝10:1)，得白色固体8(36.5mg，43.3％产率)。Under the protection of nitrogen, in an ice-water bath, a solution of acryloyl chloride (12.1 mg) in THF (1 mL) was added to compound 8-3 (73 mg), THF (4 mL), saturated Na2 CO3 (1 mL) solution, and the temperature was maintained Stir for 5min. Diluted with water, extracted three times with EA, combined the organic phases and dried with anhydrous Na2 SO4 , filtered, spin-dried, separated and purified by pre-TLC (DCM:MeOH=10:1), a white solid 8 (36.5mg, 43.3 %Yield).

ESI-MS m/z:614.40[M+H]+。ESI-MS m/z: 614.40 [M+H]+ .

实施例9：化合物9(1-(7-(8-乙氧基-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 9: Compound 9 (1-(7-(8-ethoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl )-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one) synthesis

Figure PCTCN2021124884-APPB-000048
Figure PCTCN2021124884-APPB-000048

步骤1：化合物9-1的合成Step 1: Synthesis of compound 9-1

在氮气保护下，将Sphos Pd G2(51.7mg)加入到化合物8-2(300mg)，K3PO4(228.4mg，)，乙烯基硼酸酯(110.49mg)的二氧六环(4mL)，H2O(1mL)混合溶液中，在100℃搅拌4h。降至室温，加水稀释，用EA萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，pre-TLC分离纯化(DCM:MeOH＝10:1)，得白色固体9-1(185mg，70.08％产率)。Under nitrogen protection, Sphos Pd G2 (51.7 mg) was added to compound 8-2 (300 mg), K3 PO4 (228.4 mg,), vinyl borate (110.49 mg) in dioxane (4 mL) , H2 O (1 mL) mixed solution, stirred at 100° C. for 4 h. Cool down to room temperature, dilute with water, extract three times with EA, combine the organic phases and dry with anhydrous Na2 SO4 , filter, spin dry, pre-TLC separation and purification (DCM:MeOH=10:1), a white solid 9- 1 (185 mg, 70.08% yield).

ESI-MS m/z:736.45[M+H]+。ESI-MS m/z: 736.45 [M+H]+ .

步骤2：化合物9-2的合成Step 2: Synthesis of Compound 9-2

在室温条件下，将化合物9-1(185mg)溶于DCM(4mL)和TFA(2mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到棕色油粗品9-2(138mg，粗品)。Compound 9-1 (185 mg) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain a brown oil crude product 9-2 (138 mg, crude product).

ESI-MS m/z:552.34[M+H]+。ESI-MS m/z: 552.34 [M+H]+ .

步骤3：化合物9的合成Step 3: Synthesis of compound 9

在氮气保护下，冰水浴中，将丙烯酰氯(22.6mg)的THF(1mL)溶液加入到化合物9-2(138mg)，THF(4mL)，饱和Na2CO3(1mL)溶液中，保持温度搅拌5min。加水稀释，用EA萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，pre-TLC分离纯化(DCM:MeOH＝10:1)，得白色固体9(62.5mg，38.2％产率)。Under nitrogen protection, add acryloyl chloride (22.6 mg) in THF (1 mL) solution to compound 9-2 (138 mg), THF (4 mL), saturated Na2 CO3 (1 mL) solution in an ice-water bath, and keep the temperature Stir for 5min. Diluted with water, extracted three times with EA, combined the organic phases and dried with anhydrous Na2 SO4 , filtered, spin-dried, pre-TLC separation and purification (DCM:MeOH=10:1), a white solid 9 (62.5mg, 38.2 %Yield).

ESI-MS m/z:606.45[M+H]+。ESI-MS m/z: 606.45 [M+H]+ .

1H NMR(500MHz,Chloroform-d)δ7.90(s,1H),7.51(s,1H),7.46(d,J＝8.5Hz,1H),7.37–7.34(m,1H),6.39(dd,J＝17.0,1.9Hz,1H),6.24(m,2H),5.71(dd,J＝10.2,1.9Hz,1H),5.62(d,J＝17.4Hz,1H),5.03(d,J＝11.0Hz,1H),4.22(m,1H),4.03(s,2H),3.93(s,2H),3.91– 3.84(m,1H),3.75(m,4H),3.00(t,J＝6.1Hz,2H),2.87(m,1H),2.35(s,3H),2.16(s,3H),2.12–2.08(m,5H),2.03(m,4H),1.33(m,4H).1 H NMR (500MHz, Chloroform-d) δ7.90(s, 1H), 7.51(s, 1H), 7.46(d, J=8.5Hz, 1H), 7.37–7.34(m, 1H), 6.39(dd ,J=17.0,1.9Hz,1H),6.24(m,2H),5.71(dd,J=10.2,1.9Hz,1H),5.62(d,J=17.4Hz,1H),5.03(d,J= 11.0Hz, 1H), 4.22(m, 1H), 4.03(s, 2H), 3.93(s, 2H), 3.91– 3.84(m, 1H), 3.75(m, 4H), 3.00(t, J＝6.1 Hz,2H),2.87(m,1H),2.35(s,3H),2.16(s,3H),2.12–2.08(m,5H),2.03(m,4H),1.33(m,4H).

实施例10：化合物10(1-(7-(6-氯-8-甲氧基-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 10: Compound 10 (1-(7-(6-chloro-8-methoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidine Synthesis of -4-yl)-quinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000049
Figure PCTCN2021124884-APPB-000049

步骤1：化合物10-1的合成Step 1: Synthesis of compound 10-1

在氮气保护下，将甲醇(275mg)加入到化合物M2(1.0g)，Cs2CO3(1.68g)的二氧六环(10mL)中，移至70℃反应5h。降到室温，缓慢加入水，析出固体，过滤，烘干得到黄色固体10-1(0.91g，粗品)。Under nitrogen protection, methanol (275 mg) was added to compound M2 (1.0 g), Cs2 CO3 (1.68 g) in dioxane (10 mL), and moved to 70° C. for 5 h. After cooling down to room temperature, water was slowly added to precipitate a solid, which was filtered and dried to obtain a yellow solid 10-1 (0.91 g, crude product).

ESI-MS m/z:594.18[M+H]+。ESI-MS m/z: 594.18 [M+H]+ .

步骤2：化合物10-2的合成Step 2: Synthesis of compound 10-2

在氮气保护下，将Pd(PPh3)4(77.69mg)加入到化合物10-1(200mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(172.5mg)，K3PO4(214.1mg，)的二氧六环(4mL)，H2O(1mL)溶液中，移至85℃反应4h。降到室温，加水稀释，用EA萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，硅胶柱分离纯化(DCM:MeOH＝10:1)，得黄色固体10-2(202mg，79.8％产率)。Under nitrogen protection, Pd(PPh3 )4 (77.69 mg) was added to compound 10-1 (200 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5 ,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (172.5 mg), K3 PO4 (214.1 mg,) in dioxane (4 mL), H2 O (1 mL) solution, moved to 85 ° C for 4 h. Cool down to room temperature, dilute with water, extract three times with EA, combine the organic phases and dry with anhydrous Na2 SO4 , filter, spin dry, and separate and purify on a silica gel column (DCM:MeOH=10:1) to obtain a yellow solid 10-2 (202 mg, 79.8% yield).

ESI-MS m/z:730.38[M+H]+。ESI-MS m/z: 730.38 [M+H]+ .

[根据细则91更正 15.02.2022]步骤3：化合物10-3的合成[Corrected under Rule 91 15.02.2022]Step 3: Synthesis of compound 10-3

在室温条件下，将化合物10-2(200mg)溶于DCM(6mL)和TFA(3mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到棕色油粗品10-3(149mg，粗品)。Compound 10-2 (200 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain a brown oil crude product 10-3 (149 mg, crude product).

ESI-MS m/z:546.27[M+H]+。ESI-MS m/z: 546.27 [M+H]+ .

步骤4：化合物10的合成Step 4: Synthesis ofCompound 10

[根据细则91更正 15.02.2022]在氮气保护下，冰水浴中，将丙烯酰氯(24.7mg)的THF(1mL)溶液加入到化合物10-3(149mg)，THF(4mL)，饱和Na2CO3(1mL)溶液中，保持温度搅拌5min。加水稀释，用EA萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，pre-TLC分离纯化(DCM:MeOH＝10:1)，得白色固体10(80.3mg，47.27％产率，96.39％纯度)。[Corrected 15.02.2022 according to Rule 91] Add a solution of acryloyl chloride (24.7 mg) in THF (1 mL) to compound 10-3 (149 mg), THF (4 mL), saturated Na2 CO3 (1mL) solution, keep the temperature and stir for 5min. Diluted with water, extracted three times with EA, combined the organic phases and dried with anhydrous Na2 SO4 , filtered, spin-dried, pre-TLC separation and purification (DCM:MeOH=10:1), a white solid 10 (80.3mg, 47.27 % yield, 96.39% purity).

ESI-MS m/z:600.35[M+H]+。ESI-MS m/z: 600.35 [M+H]+ .

实施例11：化合物11(1-(7-(8-甲氧基-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 11: Compound 11 (1-(7-(8-methoxy-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl )-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one) synthesis

Figure PCTCN2021124884-APPB-000050
Figure PCTCN2021124884-APPB-000050

步骤1：化合物11-1的合成Step 1: Synthesis of compound 11-1

在氮气保护下，将Sphos Pd G2(59.2mg)加入到化合物10-2(300mg)，K3PO4(261.5mg)，乙烯基硼酸酯(126.53mg)的二氧六环(4mL)，H2O(1mL)混合溶液中，在100℃搅拌4h。降至室温，加水稀释，用EA萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，pre-TLC分离纯化(DCM:MeOH＝10:1)，得白色固体11-1(205mg，69.13％产率)。Under nitrogen, Sphos Pd G2 (59.2 mg) was added to compound 10-2 (300 mg), K3 PO4 (261.5 mg), vinyl borate (126.53 mg) in dioxane (4 mL), In H2 O (1 mL) mixed solution, stirred at 100°C for 4 h. Cooled down to room temperature, diluted with water, extracted three times with EA, combined the organic phases and dried with anhydrous Na2 SO4 , filtered, spin-dried, pre-TLC separation and purification (DCM:MeOH=10:1) to obtain a white solid 11- 1 (205 mg, 69.13% yield).

ESI-MS m/z:722.43[M+H]+。ESI-MS m/z: 722.43 [M+H]+ .

步骤2：化合物11-2的合成Step 2: Synthesis of compound 11-2

在室温条件下，将化合物11-1(205mg)溶于DCM(6mL)和TFA(3mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到棕色油粗品11-2(152mg，粗品)。Compound 11-1 (205 mg) was dissolved in a mixed solvent of DCM (6 mL) and TFA (3 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain a brown oil crude product 11-2 (152 mg, crude product).

ESI-MS m/z:538.32[M+H]+。ESI-MS m/z: 538.32 [M+H]+ .

步骤3：化合物11的合成Step 3: Synthesis ofcompound 11

在氮气保护下，冰水浴中，将丙烯酰氯(25.4mg)的THF(1mL)溶液加入到化合物11-2(152mg)，THF(4mL)，饱和Na2CO3(1mL)溶液中，保持温度搅拌5min。加水稀释，用EA 萃取三次萃取，合并有机相并用无水Na2SO4干燥，过滤，旋干，pre-TLC分离纯化(DCM:MeOH＝10:1)，得黄色固体11(87.2mg，52.1％产率)。Under nitrogen protection, in an ice-water bath, add acryloyl chloride (25.4 mg) in THF (1 mL) to compound 11-2 (152 mg), THF (4 mL), saturated Na2 CO3 (1 mL) solution, and keep the temperature Stir for 5min. Diluted with water, extracted three times with EA, combined the organic phases and dried with anhydrous Na2 SO4 , filtered, spin-dried, separated and purified by pre-TLC (DCM:MeOH=10:1), a yellow solid 11 (87.2mg, 52.1 %Yield).

ESI-MS m/z:592.43[M+H]+。ESI-MS m/z: 592.43 [M+H]+ .

1H NMR(500MHz,Chloroform-d)δ7.90(s,1H),7.52(s,1H),7.47(d,J＝8.5Hz,1H),7.36(d,J＝8.5Hz,1H),6.39(dd,J＝17.0,2.0Hz,1H),6.22(m,2H),5.72(dd,J＝10.2,2.0Hz,1H),5.62(d,J＝17.4Hz,1H),5.03(d,J＝11.0Hz,1H),4.03(s,2H),3.93(s,2H),3.75(m,8H),3.00(m,3H),2.35(s,3H),2.14(s,3H),2.11(d,J＝7.1Hz,5H),2.04(m,4H).1 H NMR (500MHz, Chloroform-d) δ7.90(s, 1H), 7.52(s, 1H), 7.47(d, J=8.5Hz, 1H), 7.36(d, J=8.5Hz, 1H), 6.39(dd, J=17.0,2.0Hz,1H),6.22(m,2H),5.72(dd,J=10.2,2.0Hz,1H),5.62(d,J=17.4Hz,1H),5.03(d ,J＝11.0Hz,1H),4.03(s,2H),3.93(s,2H),3.75(m,8H),3.00(m,3H),2.35(s,3H),2.14(s,3H) ,2.11(d,J=7.1Hz,5H),2.04(m,4H).

实施例12：化合物12(1-(7-(2-(1-乙酰哌啶-4-基)-7-(5-甲基-1H-吲唑-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬-2-基)丙-2-烯-1-酮)的合成Example 12: Compound 12 (1-(7-(2-(1-acetylpiperidin-4-yl)-7-(5-methyl-1H-indazol-4-yl)-8-(2, 2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]non-2-yl)prop-2-en-1-one )Synthesis

Figure PCTCN2021124884-APPB-000051
Figure PCTCN2021124884-APPB-000051

步骤1：化合物12-1的合成Step 1: Synthesis of compound 12-1

在室温下，将1乙酰基哌啶-4-碳酰氯(567mg)加入到化合物M1(300mg)的THF(10mL)中，移至40℃搅拌4小时。待反应完全后，直接浓缩出去THF，得到黄色固体状目标产物12-1(460mg，粗品)。1 Acetylpiperidine-4-carbonyl chloride (567 mg) was added to compound M1 (300 mg) in THF (10 mL) at room temperature, and the mixture was moved to 40°C and stirred for 4 hours. After the reaction was complete, the THF was concentrated directly to obtain the target product 12-1 (460 mg, crude product) as a yellow solid.

ESI-MS m/z:512.14[M+H]+。ESI-MS m/z: 512.14 [M+H]+ .

步骤2：化合物12-2的合成Step 2: Synthesis of compound 12-2

在室温下，将甲醇钠(224.56mg)加入到化合物12-1(460mg)的toluene(10mL)中，移至110℃回流搅拌8小时。待反应完全后，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机相经Na2SO4干燥且真空浓缩，得到白色固体状目标产物12-2(210mg，粗品)。Sodium methoxide (224.56 mg) was added to toluene (10 mL) of compound 12-1 (460 mg) at room temperature, and the mixture was moved to 110° C. under reflux and stirred for 8 hours. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo to give the target product 12-2 (210 mg, crude) as a white solid.

ESI-MS m/z:494.06[M+H]+。ESI-MS m/z: 494.06 [M+H]+ .

步骤3：化合物12-3的合成Step 3: Synthesis of compound 12-3

在室温下，将化合物12-2(170mg)加到DMF(5mL)中，接着加入2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(92mg)、BOP(376mg)、DBU(258mg)，40度反应6h。反应到后期原料剩30％不再进行，向反应物中加入水，用乙酸乙酯萃取，有机相经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(展开剂为DCM/MeOH＝15:1)纯化,得到黄色固体产物12-3(150mg，62.2％收率)。Compound 12-2 (170 mg) was added to DMF (5 mL) at room temperature, followed by tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (92 mg), BOP (376 mg ), DBU (258mg), react at 40 degrees for 6h. The reaction was not carried out until 30% of the raw materials remained in the late stage. Water was added to the reactant, extracted with ethyl acetate, the organic phase was dried over Na2 SO4 and concentrated in vacuo, and the concentrate was passed through pre-TLC (developing agent: DCM/MeOH= 15:1) was purified to obtain yellow solid product 12-3 (150 mg, 62.2% yield).

ESI-MS m/z:702.28[M+H]+。ESI-MS m/z: 702.28 [M+H]+ .

步骤4：化合物12-4的合成Step 4: Synthesis of compound 12-4

在氮气保护下，将三氟乙醇(106mg)加入到化合物12-3(150mg)，Cs2CO3(139mg，)的二氧六环(3mL)中，移至100℃反应3h。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(展开剂为DCM/MeOH＝15:1)纯化,得到黄色油状的所要目标产物12-4(120mg，71.9％收率)。Under nitrogen protection, trifluoroethanol (106 mg) was added to compound 12-3 (150 mg), Cs2 CO3 (139 mg,) in dioxane (3 mL), and moved to 100° C. for 3 h. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was purified by pre-TLC (developing solvent: DCM/MeOH=15:1), The desired target product 12-4 was obtained as a yellow oil (120 mg, 71.9% yield).

ESI-MS m/z:782.48[M+H]+。ESI-MS m/z: 782.48 [M+H]+ .

步骤5：化合物12-5的合成Step 5: Synthesis of compound 12-5

在氮气保护下，将Pd(dppf)Cl2.CH2Cl2(12mg)加入到化合物12-4(115mg)，K2CO3(41mg，)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(22.7mg)的二氧六环(2.5mL)，H2O(0.5mL)混合溶液中，在70℃搅拌6h。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，剩余物通过快速硅胶柱色谱(PE:EA＝1:2)纯化，得到黄色固体状的所要目标产物12-5(65mg，65.4％产率)。Under nitrogen protection, Pd(dppf)Cl2 .CH2 Cl2 (12 mg) was added to compound 12-4 (115 mg), K2 CO3 (41 mg,), 4,4,5,5-tetramethyl - In a mixed solution of 2-vinyl-1,3,2-dioxaborolane (22.7mg) in dioxane (2.5mL), H2 O (0.5mL), stir at 70°C for 6h . The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the residue was subjected to flash silica gel column chromatography (PE:EA=1: 2) Purification afforded the desired target product 12-5 (65 mg, 65.4% yield) as a yellow solid.

ESI-MS m/z:682.58[M+H]+。ESI-MS m/z: 682.58 [M+H]+ .

步骤6：化合物12-6的合成Step 6: Synthesis of compound 12-6

在氮气保护下，将Pd(PPh3)4(22mg)加入到化合物12-5(65mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(65mg)，K3PO4(40mg)的二氧六环(2mL),H2O(0.5mL)溶液中，移至85℃反应反应约4小时。将反应混合物冷却至室温，向反应混合 物中加入水，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(PE:EA＝2:1)纯化，得到呈黄色油状的所要产物12-6(70mg，89.7％产率)。Under nitrogen protection, Pd(PPh3 )4 (22 mg) was added to compound 12-5 (65 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (65 mg), K3 PO4 (40 mg) in dioxane (2 mL), H2 O (0.5 mL ) solution, moved to 85 ° C for about 4 hours of reaction. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (PE:EA=2:1 ) to give the desired product 12-6 (70 mg, 89.7% yield) as a yellow oil.

ESI-MS m/z:818.94[M+H]+。ESI-MS m/z: 818.94 [M+H]+ .

步骤7：化合物12-7的合成Step 7: Synthesis of compound 12-7

在室温下，将化合物14-6(70mg，85.7μmol)溶于DCM(1mL)和TFA(0.5mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色固体状目标产物12-7(93mg，粗品)。Compound 14-6 (70 mg, 85.7 μmol) was dissolved in a mixed solvent of DCM (1 mL) and TFA (0.5 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the target product 12-7 (93 mg, crude product) as a yellow solid.

ESI-MS m/z:634.32[M+H]+。ESI-MS m/z: 634.32 [M+H]+ .

步骤8：化合物12的合成Step 8: Synthesis ofcompound 12

在氮气保护下，冰水浴中，将丙烯酰氯(8.4mg，94μmol)的THF溶液加入到化合物14-7(54mg)，THF(2mL)，饱和Na2CO3(0.5mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物12(25.7mg，43.2％产率)。Under nitrogen protection, in an ice-water bath, a THF solution of acryloyl chloride (8.4 mg, 94 μmol) was added to compound 14-7 (54 mg), THF (2 mL), saturated Na2 CO3 (0.5 mL) solution, and then The response is complete. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product 12 (25.7 mg, 43.2% yield) as a yellow solid.

ESI-MS m/z:688.49[M+H]+。ESI-MS m/z: 688.49 [M+H]+ .

Figure PCTCN2021124884-APPB-000052
Figure PCTCN2021124884-APPB-000052

Figure PCTCN2021124884-APPB-000053
Figure PCTCN2021124884-APPB-000053

Figure PCTCN2021124884-APPB-000054
Figure PCTCN2021124884-APPB-000054

Figure PCTCN2021124884-APPB-000055
Figure PCTCN2021124884-APPB-000055

Figure PCTCN2021124884-APPB-000056
Figure PCTCN2021124884-APPB-000056

实施例13：化合物13(1-(7-(6-环丙基-8-(2-甲氧基乙氧基)-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 13: Compound 13 (1-(7-(6-cyclopropyl-8-(2-methoxyethoxy)-7-(5-methyl-1H-indazol-4-yl)- 2-(1-Methylpiperidin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one )Synthesis

Figure PCTCN2021124884-APPB-000057
Figure PCTCN2021124884-APPB-000057

步骤1：化合物13-1的合成Step 1: Synthesis of compound 13-1

在氮气保护下，将Pd(dppf)2Cl2DCM(112mg)加入到碳酸钾(378mg),化合物1-6(1g)和环丙基硼酸(124mg)的甲苯(10mL)和水(1mL)的溶液中,100度搅拌3小时，TLC和LCMS显示反应完毕，冷却至室温，乙酸乙酯稀释，饱和食盐水洗涤两次，干燥，过滤，浓缩得粗产物，柱层析(DCM/MeOH＝15:1)得目标产物(530mg,60％产率)。ESI-MS m/z:668.30[M+H]+Under nitrogen, Pd(dppf)2 Cl2 DCM (112 mg) was added to potassium carbonate (378 mg), compound 1-6 (1 g) and cyclopropylboronic acid (124 mg) in toluene (10 mL) and water (1 mL) In the solution, stirred at 100 degrees for 3 hours, TLC and LCMS showed that the reaction was complete, cooled to room temperature, diluted with ethyl acetate, washed twice with saturated brine, dried, filtered, concentrated to give a crude product, column chromatography (DCM/MeOH= 15:1) to obtain the target product (530 mg, 60% yield). ESI-MS m/z:668.30[M+H]+

步骤2：化合物13-2的合成Step 2: Synthesis of compound 13-2

在氮气保护下，将Pd2(dba)3(9.9mg)和SPhos(8.8mg)加入到13-1(72mg),5-甲基-1-(四氢-2H-吡喃-2-基)-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1H-吲唑(37mg)和磷酸钾(46mg)的二氧六环(3mL)和水(0.4mL)的溶液中，加热至110度反应2小时，TLC和LCMS显示反应完毕，冷却至室温，浓缩，柱层析(DCM/MeOH＝15:1)得产物(38mg,45％收率)。ESI-MS m/z:804.50[M+H]+Under nitrogen protection, Pd2 (dba)3 (9.9 mg) and SPhos (8.8 mg) were added to 13-1 (72 mg), 5-methyl-1-(tetrahydro-2H-pyran-2-yl )-4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (37mg) and potassium phosphate (46mg) Hexacyclic (3mL) and water (0.4mL) solution, heated to 110 degrees for 2 hours, TLC and LCMS showed that the reaction was complete, cooled to room temperature, concentrated, column chromatography (DCM/MeOH=15:1) to obtain the product (38 mg, 45% yield). ESI-MS m/z:804.50[M+H]+

步骤3：化合物13-3的合成Step 3: Synthesis of compound 13-3

在室温下，将13-2(200mg)溶于DCM(4mL)和TFA(2mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色固体状粗品目标产物13-3(100mg)。ESI-MS m/z:620.38[M+H]+。13-2 (200 mg) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product 13-3 (100 mg) as a yellow solid. ESI-MS m/z: 620.38 [M+H]+.

步骤4：化合物13的合成Step 4: Synthesis of compound 13

在氮气保护下，冰水浴中，将丙烯酰氯(22mg)的THF溶液加入到13-3(100mg)的THF(4mL)和饱和Na2CO3(1mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物13(24.1mg，22％产率)。ESI-MS m/z:674.40[M+H]+。Under the protection of nitrogen, the THF solution of acryloyl chloride (22 mg) was added to the THF (4 mL) and saturated Na2CO3 (1 mL) solution of 13-3 (100 mg) in an ice-water bath, and the reaction was complete upon addition. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product 13 (24.1 mg, 22% yield) as a yellow solid. ESI-MS m/z: 674.40 [M+H]+.

实施例14：化合物14(1-(7-(6-环丙基-8-(2-甲氧基乙氧基)-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 14: Compound 14 (1-(7-(6-cyclopropyl-8-(2-methoxyethoxy)-7-(5-methyl-1H-indazol-4-yl)- 2-(1-Methylpiperidin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one )Synthesis

Figure PCTCN2021124884-APPB-000058
Figure PCTCN2021124884-APPB-000058

在氮气保护下，将乙二醇甲醚(762mg)加入到1-5(1.35g)，Cs2CO3(1.96g)的Dioxane(15mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯/甲醇＝10:1萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物14-1(1.3g，粗产物)。ESI-MS m/z:730.15[M+H]+Under nitrogen protection, ethylene glycol methyl ether (762 mg) was added to 1-5 (1.35 g), Cs2 CO3 (1.96 g) in Dioxane (15 mL), and moved to 100° C. for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product 14-1 as a yellow solid (1.3 g , crude product). ESI-MS m/z:730.15[M+H]+

之后按照化合物13的合成步骤合成14-2，14-3和14-4，最终得到目标产物14。ESI-MS m/z:650.4[M+H]+Afterwards, 14-2, 14-3 and 14-4 were synthesized according to the synthesis steps of compound 13, and finally thetarget product 14 was obtained. ESI-MS m/z:650.4[M+H]+

实施例15：化合物15(1-(7-(6-环丙基-8-(2,2-二氟乙氧基)-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)喹唑啉-4–基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 15: Compound 15 (1-(7-(6-cyclopropyl-8-(2,2-difluoroethoxy)-7-(5-methyl-1H-indazol-4-yl) -2-(1-Methylpiperidin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1- ketone) synthesis

Figure PCTCN2021124884-APPB-000059
Figure PCTCN2021124884-APPB-000059

步骤1：化合物15-1的合成Step 1: Synthesis of compound 15-1

在氮气保护下，将二氟乙醇(201mg)加入到1-5(550mg)，Cs2CO3(531.45mg)的Dioxane(5mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯/甲醇＝10:1萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物15-1(550mg，crude)。ESI-MS m/z:736.30[M+H]+Under nitrogen protection, difluoroethanol (201 mg) was added to 1-5 (550 mg), Cs2 CO3 (531.45 mg) in Dioxane (5 mL), and moved to 100° C. for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product 15-1 (550 mg, cruel). ESI-MS m/z:736.30[M+H]+

之后按照化合物13的合成步骤合成15-2，15-3和15-4，最终得到目标产物15。ESI-MS m/z:656.4[M+H]+Afterwards, 15-2, 15-3 and 15-4 were synthesized according to the synthesis steps of compound 13, and finally thetarget product 15 was obtained. ESI-MS m/z:656.4[M+H]+

Figure PCTCN2021124884-APPB-000060
Figure PCTCN2021124884-APPB-000060

Figure PCTCN2021124884-APPB-000061
Figure PCTCN2021124884-APPB-000061

实施例16：化合物16(1-(7-(8-乙氧基-6-乙基-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)喹唑啉-4-基)-2，7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 16: Compound 16 (1-(7-(8-ethoxy-6-ethyl-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiper Synthesis of pyridin-4-yl)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000062
Figure PCTCN2021124884-APPB-000062

步骤1：化合物16-1的合成Step 1: Synthesis of compound 16-1

在氮气保护下，将乙醇(112mg)加入到1-5(550mg),Cs2CO3(531mg)的Dioxane(5mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯/甲醇＝10:1萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到黄色固体状的所要目标产物16-1(590mg，crude)。ESI-MS m/z:700.3[M+H]+。Under nitrogen protection, ethanol (112 mg) was added to 1-5 (550 mg), Cs2 CO3 (531 mg) in Dioxane (5 mL), and moved to 100° C. for 30 min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate/methanol = 10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product 16-1 (590 mg, cruel). ESI-MS m/z: 700.3 [M+H]+.

步骤2：化合物16-2的合成Step 2: Synthesis of compound 16-2

在氮气保护下，将Pd(dppf)Cl2.CH2Cl2(67mg)加入到16-1(粗品590mg)，K2CO3(226mg)，4,4,5,5-四甲基-2-乙烯基-1,3,2-二氧杂硼杂环戊烷(126mg)的dioxane(5.0mL)，H2O(0.5mL)混合溶液中，在70℃搅拌1h。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物16-2(329mg，66％产率).ESI-MS m/z:600.6[M+H]+。Under nitrogen protection, Pd(dppf)Cl2 .CH2 Cl2 (67 mg) was added to 16-1 (crude product 590 mg), K2 CO3 (226 mg), 4,4,5,5-tetramethyl- A mixed solution of 2-vinyl-1,3,2-dioxaborolane (126 mg) in dioxane (5.0 mL) and H2 O (0.5 mL) was stirred at 70°C for 1 h. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1) to obtain the desired product 16-2 (329 mg, 66% yield) as a yellow solid. ESI-MS m/z: 600.6 [M+H]+.

步骤3：化合物16-3的合成Step 3: Synthesis of compound 16-3

在氮气保护下，将Pd(PPh3)4(116.16mg)加入到16-3(329mg)，5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(344.04mg)，K3PO4(213.38mg)的dioxane(3mL),H2O(0.75mL)溶液中，移至85℃反应约3小时。将反应混合物冷却至室温，向反应混合物中加入水，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物16-3(244mg，65％产率)。ESI-MS m/z:736.60[M+H]+。Under nitrogen protection, Pd(PPh3 )4 (116.16 mg) was added to 16-3 (329 mg), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (344.04 mg), K3 PO4 (213.38 mg) in dioxane (3 mL), H2 O (0.75 mL) solution, moved to 85 ° C for about 3 hours. The reaction mixture was cooled to room temperature, water was added to the reaction mixture, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed through pre-TLC (DCM /MeOH=10:1) to obtain the desired product 16-3 (244 mg, 65% yield) as a yellow solid. ESI-MS m/z: 736.60 [M+H]+.

步骤4：化合物16-4的合成Step 4: Synthesis of Compound 16-4

在室温下，将16-3(100mg)溶于甲醇，加入钯碳300mg，置换氢气，移至40℃反应过夜。待反应完全后，硅藻土助滤，滤液浓缩除去溶剂，得到黄色固体状粗品目标产物16-4(150mg，crude)。ESI-MS m/z:738.60[M+H]+。At room temperature, 16-3 (100 mg) was dissolved in methanol, 300 mg of palladium carbon was added to replace the hydrogen, and the reaction was carried out at 40° C. overnight. After the reaction was complete, Celite was used to filter, and the filtrate was concentrated to remove the solvent to obtain the crude target product 16-4 (150 mg, crude) as a yellow solid. ESI-MS m/z: 738.60 [M+H]+.

步骤5：化合物16-5的合成Step 5: Synthesis of compound 16-5

在室温下，将16-4(150mg,crude)溶于DCM(4mL)和TFA(2mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得到黄色固体状粗品目标产物16-5(100mg，crude)。ESI-MS m/z:554.60[M+H]+。16-4 (150 mg, crude) was dissolved in a mixed solvent of DCM (4 mL) and TFA (2 mL) at room temperature, and moved to 40 °C for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product 16-5 (100 mg, crude) as a yellow solid. ESI-MS m/z: 554.60 [M+H]+.

步骤6：化合物16的合成Step 6: Synthesis of compound 16

在氮气保护下，冰水浴中，将丙烯酰氯(17.03mg)的THF溶液加入到16-5(100mg)，THF(4mL)，饱和Na2CO3(1mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯:甲醇＝10:1萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到呈黄色固体状的所要产物16(21mg，20％产率)。ESI-MS m/z:608.40[M+H]+。Under nitrogen protection, in an ice-water bath, a THF solution of acryloyl chloride (17.03 mg) was added to 16-5 (100 mg), THF (4 mL), saturated Na2CO3 (1 mL) solution, and the reaction was complete upon addition. The reaction mixture was poured into water, extracted with ethyl acetate:methanol=10:1, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was passed by pre-TLC (DCM/MeOH=10:1) Purification afforded the desired product 16 (21 mg, 20% yield) as a yellow solid. ESI-MS m/z: 608.40 [M+H]+.

实施例17-22分别采用环丁醇，异丙醇，二氟乙醇，2-氟乙醇，乙二醇甲醚和环丙甲醇与化合物1-5反应得到相应的产物，然后按照16的合成步骤合成相应的最终产物化合物17-22。Examples 17-22 respectively adopt cyclobutanol, isopropanol, difluoroethanol, 2-fluoroethanol, ethylene glycol methyl ether and cyclopropylmethanol to react with compound 1-5 to obtain corresponding products, and then follow the synthesis steps of 16 The corresponding final products compounds 17-22 were synthesized.

Figure PCTCN2021124884-APPB-000063
Figure PCTCN2021124884-APPB-000063

Figure PCTCN2021124884-APPB-000064
Figure PCTCN2021124884-APPB-000064

实施例23：化合物23(1-(7-(6-氯-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-8-(2,2,2-三氟乙氧基)喹唑啉-4-yl)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 23: Compound 23 (1-(7-(6-chloro-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl)- 8-(2,2,2-trifluoroethoxy)quinazoline-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1- ketone) synthesis

Figure PCTCN2021124884-APPB-000065
Figure PCTCN2021124884-APPB-000065

步骤1：化合物23-1的合成Step 1: Synthesis of compound 23-1

在室温下,向2-氨基-4-溴-3-氟苯甲酸(20g)于DMF(200mL)中的溶液中添加NCS(13.8g)且所得混合物在70℃搅拌过夜。反应完全，将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩得褐色固体状的所要粗品产物(22g,98％产率)。ESI-MS m/z:267.10[M+H]+。To a solution of 2-amino-4-bromo-3-fluorobenzoic acid (20 g) in DMF (200 mL) was added NCS (13.8 g) at room temperature and the resulting mixture was stirred at 70 °C overnight. After the reaction was complete, the mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo to give the desired crude product (22 g, 98% yield) as a brown solid. ESI-MS m/z: 267.10 [M+H]+.

步骤2：化合物23-2的合成Step 2: Synthesis of Compound 23-2

在室温下，将1-1(2.42g)加入到M1-5(2.33g)的THF(20mL)中，移至40℃搅拌过夜。待反应完全后，加入氨水30mL，调节溶液PH至11，35度搅拌，检测直至反应完全，有大量不溶物析出，过滤得不溶物，即粗品23-2(3.5g，crude)。ESI-MS m/z:374.2[M+H]+。1-1 (2.42 g) was added to THF (20 mL) of M1-5 (2.33 g) at room temperature, and the mixture was moved to 40° C. and stirred overnight. After the reaction is complete, add 30 mL of ammonia water, adjust the pH of the solution to 11, stir at 35°C, and check until the reaction is complete. A large amount of insoluble matter precipitates out, and the insoluble matter is obtained by filtration, namely crude product 23-2 (3.5 g, crude). ESI-MS m/z: 374.2 [M+H]+.

步骤3：化合物23-3的合成Step 3: Synthesis of Compound 23-3

室温氮气保护下，在反应瓶中加入化合物23-2(1g)和2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(725mg)，加入DMF(10mL)和DBU(2.44g)，冰水浴降温，加入BOP(2.95g)，升温至45度反应2小时,检测反应完毕，冷却至室温，乙酸乙酯稀释，水稀释，水相乙酸乙酯萃取一次，有机相合并，0.5N盐酸和Brine(1:1)洗涤3次，aq.NaHCO3洗涤一次，食盐水洗涤一次，硫酸钠干燥，浓缩得目标产物23-3(1.2g，粗产物)。Under the protection of nitrogen at room temperature, compound 23-2 (1g) and tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (725mg) were added to the reaction flask, DMF (10mL) and DBU (2.44g), cool down in an ice-water bath, add BOP (2.95g), heat up to 45 degrees and react for 2 hours, check that the reaction is complete, cool to room temperature, dilute with ethyl acetate, dilute with water, extract the aqueous phase with ethyl acetate once, and the organic phase Combined, washed 3 times with 0.5N hydrochloric acid and Brine (1:1), once with aq.NaHCO3 , once with brine, dried over sodium sulfate, and concentrated to obtain the target product 23-3 (1.2 g, crude product).

参照化合物13的合成步骤，用三氟乙醇碱性条件下取代氟原子合成23-4,23-4用Suzuki偶联的方法合成23-5,23-5在三氟乙酸作用下脱掉保护基得到化合物23-6,23-6在碱性条件下与丙烯酰氯反应合成目标产物23.Referring to the synthesis steps of compound 13, use trifluoroethanol to replace the fluorine atom under basic conditions to synthesize 23-4, 23-4, synthesize 23-5, 23-5 by Suzuki coupling method, and remove the protecting group under the action of trifluoroacetic acid The compound 23-6 was obtained, and the target product 23 was synthesized by reacting 23-6 with acryloyl chloride under basic conditions.

参照化合物23的合成方法完成化合物24和25的合成，24和25分别用环丁醇和二氟乙醇代替三氟乙醇取代氟原子得到相应的产物，之后按照23的步骤合成目标产物24和25。The synthesis ofcompounds 24 and 25 was completed by referring to the synthesis method of compound 23. Cyclobutanol and difluoroethanol were used to replace trifluoroethanol in 24 and 25, respectively, to obtain the corresponding products, and then thetarget products 24 and 25 were synthesized according to the steps of 23.

Figure PCTCN2021124884-APPB-000066
Figure PCTCN2021124884-APPB-000066

Figure PCTCN2021124884-APPB-000067
Figure PCTCN2021124884-APPB-000067

实施例26：化合物26(1-(7-(6-乙基-2-(1-(2-甲氧基乙基)哌啶-4-基)-7-(5-甲基-1H-吲唑-4-基)-8-(2,2，2-三氟乙氧基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 26: Compound 26 (1-(7-(6-ethyl-2-(1-(2-methoxyethyl)piperidin-4-yl)-7-(5-methyl-1H- Indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl) Synthesis of prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000068
Figure PCTCN2021124884-APPB-000068

步骤1：化合物26-1的合成Step 1: Synthesis of compound 26-1

室温下，将哌啶-4-羧酸(7.000g)加入THF(200mL)和水(40mL)中，加入三乙胺(16.45g)，加入(2,5-二氧杂吡咯烷-1-基)2-三甲基甲硅烷基乙基碳酸酯(15.46g)，室温搅拌至反应完毕，浓缩掉THF，降温，1N HCl酸化至PH2-3左右，DCM萃取三次，有机相盐水洗两次，干燥，浓缩得目标产物26-1(16g,粗品)。At room temperature, add piperidine-4-carboxylic acid (7.000g) into THF (200mL) and water (40mL), add triethylamine (16.45g), add (2,5-dioxapyrrolidine-1- Base) 2-trimethylsilyl ethyl carbonate (15.46g), stirred at room temperature until the reaction was completed, concentrated THF, cooled down, acidified to about PH2-3 with 1N HCl, extracted three times with DCM, washed the organic phase twice with brine , dried, and concentrated to obtain the target product 26-1 (16 g, crude product).

步骤2：化合物26-2的合成Step 2: Synthesis of compound 26-2

室温下，将化合物26-1(2.73g)加入DCM(30mL)中，加入2滴DMF,滴入草酰氯(3.81g)，有气泡冒出，反应完毕，浓缩得26-2(3g,粗产物)，直接用于下一步。At room temperature, compound 26-1 (2.73g) was added to DCM (30mL), 2 drops of DMF was added, oxalyl chloride (3.81g) was added dropwise, bubbles appeared, the reaction was completed, and concentrated to give 26-2 (3g, crude product), which was used directly in the next step.

步骤3：化合物26-3和26-4的合成Step 3: Synthesis of Compounds 26-3 and 26-4

在室温下，将26-2(3g)加入到M1(1.8g)的THF(40mL)中，移至40℃搅拌过夜，检测直至反应完全后得26-3，加入氨水30mL，调节溶液PH至11，40度搅拌，检测直至反应完全，有大量不溶物析出，过滤得不溶物，即26-4(1.9g，63％收率)。ESI-MS m/z:374.2[M+H]+。At room temperature, add 26-2 (3g) into THF (40mL) of M1 (1.8g), move to 40°C and stir overnight, check until the reaction is complete to obtain 26-3, add ammonia water 30mL, adjust the pH of the solution to 11. Stir at 40°C and check until the reaction is complete. A large amount of insoluble matter is precipitated, and the insoluble matter is obtained by filtration, namely 26-4 (1.9 g, 63% yield). ESI-MS m/z: 374.2 [M+H]+.

步骤4：化合物26-5的合成Step 4: Synthesis of Compound 26-5

室温氮气保护下，在反应瓶中加入化合物26-4(1.22g)和2,7-二氮杂螺[3.5]壬烷-2-羧酸叔丁酯(695mg)，加入DMF(12mL)和DBU(1.87g)，冰水浴降温，加入BOP(2.2g)，升温 至45度反应2小时,检测反应完毕，冷却至室温，乙酸乙酯稀释，水稀释，水相乙酸乙酯萃取一次，有机相合并，0.5N盐酸和Brine(1:1)洗涤3次，aq.NaHCO3洗涤一次，Brine洗涤一次，硫酸钠干燥，浓缩得目标产物26-5(1g，粗产物)。Under the protection of nitrogen at room temperature, compound 26-4 (1.22g) and tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylate (695mg) were added to the reaction flask, DMF (12mL) and DBU (1.87g), cool down in an ice-water bath, add BOP (2.2g), heat up to 45 degrees and react for 2 hours, check that the reaction is complete, cool to room temperature, dilute with ethyl acetate, dilute with water, extract the aqueous phase with ethyl acetate once, organic The phases were combined, washed 3 times with 0.5N hydrochloric acid and Brine (1:1), washed once with aq.NaHCO3 , washed once with Brine, dried over sodium sulfate, and concentrated to obtain the target product 26-5 (1 g, crude product).

步骤5：化合物26-6的合成Step 5: Synthesis of Compound 26-6

在氮气保护下，将三氟乙醇(2.77g)加入到26-5(2.23g),Cs2CO3(2.7g)的Dioxane(25mL)中，移至100℃反应30min。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，得到固体状的所要目标产物26-6(4.6g，crude)。ESI-MS m/z:784.30[M+H]+Under nitrogen protection, trifluoroethanol (2.77g) was added to 26-5 (2.23g), Cs2 CO3 (2.7g) in Dioxane (25mL), moved to 100°C for 30min. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give the desired target product 26-6 as a solid (4.6 g, crude). ESI-MS m/z:784.30[M+H]+

步骤6：化合物26-7的合成Step 6: Synthesis of compound 26-7

在氮气保护下，将Pd(dppf)2Cl2DCM(1.82g)加入到碳酸钾(6.16g),化合物26-6(19.7g)和乙烯基硼酸酯(3.43g)的二氧六环(200mL)和水(20mL)的溶液中,100度搅拌3小时，TLC和LCMS显示反应完毕，冷却至室温，乙酸乙酯稀释，饱和食盐水洗涤两次，干燥，过滤，浓缩得粗产物，柱层析得(DCM/EA＝20:1)目标产物(9.6g,55％产率)。ESI-MS m/z:895.33[M+H]+Under nitrogen, Pd(dppf)2 Cl2 DCM (1.82 g) was added to potassium carbonate (6.16 g), compound 26-6 (19.7 g) and vinyl borate (3.43 g) in dioxane (200mL) and water (20mL), stirred at 100°C for 3 hours, TLC and LCMS showed that the reaction was complete, cooled to room temperature, diluted with ethyl acetate, washed twice with saturated brine, dried, filtered, and concentrated to give the crude product, The target product (9.6 g, 55% yield) was obtained by column chromatography (DCM/EA=20:1). ESI-MS m/z:895.33[M+H]+

步骤7：化合物26-8的合成Step 7: Synthesis of Compound 26-8

在氮气保护下，将Pd(PPh3)4(2.94g)加入到D1-7(10g),5-甲基-1-四氢吡喃-2-基-4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吲唑(7.85g)和磷酸钾(5.41g)的二氧六环(100mL)和水(20mL)的溶液中，加热至85度反应4小时，TLC和LCMS显示反应完毕，冷却至室温，乙酸乙酯稀释，水洗，饱和食盐水洗涤，干燥浓缩得粗品，柱层析得黄色泡沫状固体(10g，85％收率)。ESI-MS m/z:920.40[M+H]+Under nitrogen protection, Pd(PPh3 )4 (2.94g) was added to D1-7 (10g), 5-methyl-1-tetrahydropyran-2-yl-4-(4,4,5, A solution of 5-tetramethyl-1,3,2-dioxaborolan-2-yl)indazole (7.85 g) and potassium phosphate (5.41 g) in dioxane (100 mL) and water (20 mL) , heated to 85 degrees to react for 4 hours, TLC and LCMS showed that the reaction was complete, cooled to room temperature, diluted with ethyl acetate, washed with water, washed with saturated brine, dried and concentrated to obtain a crude product, and column chromatography gave a yellow foamy solid (10g, 85 % yield). ESI-MS m/z:920.40[M+H]+

步骤8：化合物26-9的合成Step 8: Synthesis of compound 26-9

在室温下，将26-8(1g)溶于甲醇，加入钯碳(1g)，置换氢气，移至40℃反应过夜。待反应完全后，硅藻土助滤，滤液浓缩除去溶剂，得到黄色固体状粗品目标产物26-9(1g，crude)。ESI-MS m/z:922.50[M+H]+。At room temperature, 26-8 (1 g) was dissolved in methanol, palladium carbon (1 g) was added to replace the hydrogen, and the mixture was moved to 40° C. to react overnight. After the reaction was complete, Celite was used to filter, and the filtrate was concentrated to remove the solvent to obtain the crude target product 26-9 (1 g, crude) as a yellow solid. ESI-MS m/z: 922.50 [M+H]+.

步骤9：化合物26-10的合成Step 9: Synthesis of compound 26-10

室温下，将化合物26-9(1.6g)溶于THF(10mL)，加入TBAF(4mL，1M in THF)，35度搅拌4小时，浓缩，乙酸乙酯稀释，水洗2次，饱和食盐水洗涤两次，硫酸钠干燥，浓缩得产品(912mg，67％收率)。Dissolve compound 26-9 (1.6g) in THF (10mL) at room temperature, add TBAF (4mL, 1M in THF), stir at 35°C for 4 hours, concentrate, dilute with ethyl acetate, wash twice with water, and wash with saturated brine Twice, dried over sodium sulfate, concentrated to give the product (912 mg, 67% yield).

步骤10：化合物26-11的合成Step 10: Synthesis of compound 26-11

室温下将化合物26-10(100mg)溶于DMF(5mL)，加入碳酸铯(126mg)，加入 2-溴乙基甲基醚(54mg)室温搅拌过夜，LCMS监控反应完毕，乙酸乙酯稀释，水洗一次，饱和食盐水洗涤两次，干燥，浓缩得粗品，制备板(DCM/MeOH＝10:1)制备得26-11(37mg，34％收率)ESI-MS m/z:836.46[M+H]+Compound 26-10 (100 mg) was dissolved in DMF (5 mL) at room temperature, cesium carbonate (126 mg) was added, 2-bromoethyl methyl ether (54 mg) was added and stirred at room temperature overnight, LCMS monitored the completion of the reaction, diluted with ethyl acetate, Washed once with water, washed twice with saturated brine, dried, and concentrated to obtain the crude product, the preparation plate (DCM/MeOH=10:1) prepared 26-11 (37 mg, 34% yield) ESI-MS m/z: 836.46 [M +H]+

步骤11：化合物26-12的合成Step 11: Synthesis of compound 26-12

在室温下，将26-11(37mg)溶于DCM(2mL)和TFA(1mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得粗品目标产物26-12。ESI-MS m/z:652.40[M+H]+。26-11 (37 mg) was dissolved in a mixed solvent of DCM (2 mL) and TFA (1 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product 26-12. ESI-MS m/z: 652.40 [M+H]+.

步骤12：化合物26的合成Step 12: Synthesis of compound 26

在氮气保护下，冰水浴中，将丙烯酰氯(4mg)的THF溶液加入到26-12(29mg，粗品)的THF(2mL)和饱和Na2CO3(0.5mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到所要产物26(3.1mg，6％产率)。ESI-MS m/z:706.45[M+H]+。Under the protection of nitrogen, add the THF solution of acryloyl chloride (4 mg) to the THF (2 mL) and saturated Na2 CO3 (0.5 mL) solution of 26-12 (29 mg, crude product) in an ice-water bath, and the reaction is complete upon addition . The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was purified by pre-TLC (DCM/MeOH=10:1) to give the desired product 26 (3.1 mg, 6% yield). ESI-MS m/z: 706.45 [M+H]+.

实施例27：化合物27(1-(7-(6-乙基-2-(1-(2-羟基-2-甲基丙基)哌啶-4-基)-7-(5-甲基-1H-吲唑-4-基)-8-(2,2,2-三氟乙氧基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 27: Compound 27 (1-(7-(6-ethyl-2-(1-(2-hydroxyl-2-methylpropyl)piperidin-4-yl)-7-(5-methyl -1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2 Synthesis of -yl)prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000069
Figure PCTCN2021124884-APPB-000069

步骤1：化合物27-11的合成Step 1: Synthesis of compound 27-11

将化合物27-10(100mg，0.13mmol)溶于乙醇(2mL)，加入甲基环氧丙烷(93mg，)和三乙胺(40mg)，100度微波反应0.5小时，LCMS检测反应完毕，直接浓缩得粗品27-11。Dissolve compound 27-10 (100mg, 0.13mmol) in ethanol (2mL), add propylene oxide (93mg,) and triethylamine (40mg), react in microwave at 100 degrees for 0.5 hours, LCMS detects that the reaction is complete, and concentrate directly The crude product 27-11 was obtained.

参照实施例26的合成方法在三氟乙酸作用下脱去保护基，然后碱性条件下与丙烯酰氯反应得目标产物27(8.8mg)。ESI-MS m/z:720.35[M+H]+Referring to the synthesis method of Example 26, the protective group was removed under the action of trifluoroacetic acid, and then reacted with acryloyl chloride under basic conditions to obtain the target product 27 (8.8 mg). ESI-MS m/z:720.35[M+H]+

实施例28：化合物28(1-(7-(6-乙基-7-(5-甲基-1H-吲唑-4-基)-2-(1-(氧杂环丁-3-基)哌啶-4-基)-8-(2，2,2-三氟乙氧基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 28: Compound 28 (1-(7-(6-ethyl-7-(5-methyl-1H-indazol-4-yl)-2-(1-(oxetan-3-yl) )piperidin-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl ) Synthesis of prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000070
Figure PCTCN2021124884-APPB-000070

步骤1：化合物28-11的合成Step 1: Synthesis of Compound 28-11

将化合物26-10(165mg)溶于1，2-二氯乙烷(10mL)，加入3-氧杂环丁酮(76.4mg，)和醋酸硼氢化钠(225mg)，室温35度搅拌过夜，反应完毕，乙酸乙酯稀释，水洗，饱和食盐水洗涤，干燥浓缩得粗品。Compound 26-10 (165 mg) was dissolved in 1,2-dichloroethane (10 mL), 3-oxetanone (76.4 mg,) and sodium acetate borohydride (225 mg) were added, and stirred at room temperature at 35 degrees overnight, After the reaction was completed, it was diluted with ethyl acetate, washed with water, washed with saturated brine, dried and concentrated to obtain a crude product.

参照实施例26的合成步骤在三氟乙酸作用下脱去保护基，然后碱性条件下与丙烯酰氯反应得目标产物28(13.5mg)。ESI-MS m/z:704.50[M+H]+Referring to the synthesis steps of Example 26, the protective group was removed under the action of trifluoroacetic acid, and then reacted with acryloyl chloride under basic conditions to obtain the target product 28 (13.5 mg). ESI-MS m/z:704.50[M+H]+

Figure PCTCN2021124884-APPB-000071
Figure PCTCN2021124884-APPB-000071

Figure PCTCN2021124884-APPB-000072
Figure PCTCN2021124884-APPB-000072

实施例29：化合物29(1-(7-(6-乙基-2-(1-(2-羟基-2-甲基丙基)氮杂环丁烷-3-基)-7-(5-甲基-1H-吲唑-4-基)-8-(2,2,2-三氟乙氧基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成。Example 29: Compound 29 (1-(7-(6-ethyl-2-(1-(2-hydroxyl-2-methylpropyl)azetidin-3-yl)-7-(5 -Methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonane Synthesis of -2-yl)prop-2-en-1-one).

Figure PCTCN2021124884-APPB-000073
Figure PCTCN2021124884-APPB-000073

参照实施例26和27的合成步骤。Refer to the synthetic steps of Examples 26 and 27.

化合物29-1操作参考化合物26-1的合成操作，将3-吖丁啶羧酸的氮原子进行保护。化合物29-2操作参考化合物26-2的合成操作，将化合物26-2的羧酸转变成酰氯，然后与化合物M1反应得到化合物29-3，化合物29-3不经过处理直接加入氨水调节pH至11关 环合成化合物29-4。化合物29-4在DBU和BOP作用下引入螺环合成化合物29-5，然后用三氟乙醇取代其氟原子得到化合物29-6，化合物29-6进行两次Suzuki偶联反应合成化合物29-8，之后双键还原，脱掉氮原子的保护基得到共用中间体29-10。化合物29-10参考27-11的合成步骤，在三乙胺和乙醇体系中与甲基环氧丙烷微波反应即可得到化合物29-11,之后三氟乙酸脱保护得到化合物29-12，在碱性条件下与丙烯酰氯反应得到化合物29。The operation of compound 29-1 refers to the synthesis operation of compound 26-1, and the nitrogen atom of 3-azetidinecarboxylic acid is protected. The operation of compound 29-2 refers to the synthesis operation of compound 26-2. The carboxylic acid of compound 26-2 is converted into an acid chloride, and then reacted with compound M1 to obtain compound 29-3. Compound 29-3 is directly added ammonia water to adjust the pH to 11 ring closure to synthesize compound 29-4. Compound 29-4 was introduced into a spiro ring under the action of DBU and BOP to synthesize compound 29-5, and then its fluorine atom was replaced by trifluoroethanol to obtain compound 29-6. Compound 29-6 was subjected to two Suzuki coupling reactions to synthesize compound 29-8 , followed by reduction of the double bond and deprotection of the nitrogen atom afforded the common intermediate 29-10. For compound 29-10, refer to the synthesis steps of 27-11. Compound 29-11 can be obtained by microwave reaction with methyl propylene oxide in triethylamine and ethanol system, and then compound 29-12 can be obtained by deprotection of trifluoroacetic acid. Compound 29 was obtained by reacting with acryloyl chloride under neutral conditions.

实施例30：化合物30(3-(3-(4-(2-丙烯酰基-2,7-二氮杂螺并[3.5]壬南-7-基)-6-乙基-7-(5-甲基-1H-吲唑-4-基)-8-(2,2,2-三氟乙氧基)喹唑啉-2-基)氮杂环丁烷-1-基)丙腈)的合成Example 30: Compound 30 (3-(3-(4-(2-acryloyl-2,7-diazaspiro[3.5]nonan-7-yl)-6-ethyl-7-(5 -Methyl-1H-indazol-4-yl)-8-(2,2,2-trifluoroethoxy)quinazolin-2-yl)azetidin-1-yl)propionitrile) Synthesis

Figure PCTCN2021124884-APPB-000074
Figure PCTCN2021124884-APPB-000074

步骤1；化合物30-11合成Step 1; Synthesis of compound 30-11

将化合物29-10(50mg)溶于乙醇(3mL)，加入丙烯腈(34mg)和三乙胺(7mg)，室温35度反应4小时，浓缩，乙酸乙酯稀释，水洗，饱和3食盐水洗涤，干燥浓缩，制备板(DCM/MeOH＝20:1)纯化得产物(26mg)。ESI-MS m/z:830.20[M+H]+Dissolve compound 29-10 (50 mg) in ethanol (3 mL), add acrylonitrile (34 mg) and triethylamine (7 mg), react atroom temperature 35 degrees for 4 hours, concentrate, dilute with ethyl acetate, wash with water, and wash with saturated brine , dried and concentrated, and purified on a preparative plate (DCM/MeOH=20:1) to obtain the product (26 mg). ESI-MS m/z:830.20[M+H]+

之后参照实施例26的合成方法在三氟乙酸条件下脱掉保护基，碱性条件下引入丙烯酰基，得到化合物30。Then referring to the synthesis method of Example 26, the protecting group was removed under the condition of trifluoroacetic acid, and the acryloyl group was introduced under basic condition to obtaincompound 30.

实施例31，33，34,35和37参考实施例28的方法合成，共用中间体29-10分别与氧杂环丁酮，四氢吡喃酮，环丁酮，环戊酮和乙醛在还原胺化的条件下反应制备相应的目标化合物，然后分别进行脱保护和引入丙烯酰基最终得到目标化合物31，33，34,35和37。Embodiments 31, 33, 34, 35 and 37 were synthesized with reference to the method of Example 28, and the common intermediate 29-10 was mixed with oxetanone, tetrahydropyrone, cyclobutanone, cyclopentanone and acetaldehyde respectively in Reductive amination conditions were used to prepare the corresponding target compounds, and then deprotection and introduction of acryloyl groups were carried out respectively to finally obtain the target compounds 31, 33, 34, 35 and 37.

实施例32的合成操作参照实施例30的合成操作，采用1，4-加成的方法，甲基乙烯砜与共用中间体29-10反应得到相应产物，然后脱保护和引入丙烯酰基最终得到目标化合物实施例32。The synthesis operation of Example 32 refers to the synthesis operation of Example 30, using the 1,4-addition method, methyl vinyl sulfone reacts with the common intermediate 29-10 to obtain the corresponding product, and then deprotects and introduces an acryloyl group to finally obtain the target Compound Example 32.

实施例36的合成步骤是：用共用中间体29-10与环丙基甲酰氯在二氯甲烷和碳酸氢钠水溶液体系中反应得到相应化合物，然后脱保护和引入丙烯酰基最终得到目标化合物实施例36。The synthesis steps of Example 36 are: use the common intermediate 29-10 to react with cyclopropylformyl chloride in dichloromethane and sodium bicarbonate aqueous solution to obtain the corresponding compound, then deprotect and introduce acryloyl group to finally obtain the target compound Example 36.

Figure PCTCN2021124884-APPB-000075
Figure PCTCN2021124884-APPB-000075

Figure PCTCN2021124884-APPB-000076
Figure PCTCN2021124884-APPB-000076

Figure PCTCN2021124884-APPB-000077
Figure PCTCN2021124884-APPB-000077

实施例38：化合物38(1-(7-(8-(2-羟基-2-甲基丙氧基)-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-6-乙烯基喹唑啉-4-yl)-2，7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 38: Compound 38 (1-(7-(8-(2-hydroxy-2-methylpropoxy)-7-(5-methyl-1H-indazol-4-yl)-2-( 1-methylpiperidin-4-yl)-6-vinylquinazoline-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-ene-1 - ketone) synthesis

Figure PCTCN2021124884-APPB-000078
Figure PCTCN2021124884-APPB-000078

实施例38的合成操作参考实施例16的操作，用2-甲基-1,2-丙二醇与共用中间体1-5反应得到化合物38-1，之后按照实施例16进行两次偶联反应，脱保护和上丙烯酰基步骤完成38的合成。The synthetic operation of Example 38 is referred to the operation of Example 16, using 2-methyl-1,2-propanediol to react with common intermediate 1-5 to obtain compound 38-1, and then perform two coupling reactions according to Example 16, The deprotection and acryloyl steps complete the synthesis of 38.

实施例39和40分别用二氟乙醇和1-(羟基甲基)环丙烷甲腈与共用中间体1-5反应得到相应的目标化合物，之后按照实施例16的步骤进行最终产物39和40的合成Examples 39 and 40 respectively use difluoroethanol and 1-(hydroxymethyl) cyclopropanecarbonitrile to react with common intermediate 1-5 to obtain the corresponding target compound, and then carry out thefinal product 39 and 40 according to the steps of Example 16 synthesis

Figure PCTCN2021124884-APPB-000079
Figure PCTCN2021124884-APPB-000079

Figure PCTCN2021124884-APPB-000080
Figure PCTCN2021124884-APPB-000080

实施例41：化合物41(1-(6-(7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,6-二氮杂螺[3.3]庚-2-基)丙-2-烯-1-酮)的合成Example 41: Compound 41 (1-(6-(7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl)-8-(2 ,2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,6-diazaspiro[3.3]hept-2-yl)prop-2-en-1- ketone) synthesis

Figure PCTCN2021124884-APPB-000081
Figure PCTCN2021124884-APPB-000081

步骤1：化合物41-1的合成Step 1: Synthesis of Compound 41-1

在室温下，将DIEA(480mg)加入到1-4(600mg)，2,6-二氮杂螺[3.3]庚烷-2-甲酸叔丁酯(245mg)的二氧六环溶液(10mL)中，室温40度搅拌。将混合物倾注入冰水中，用乙酸乙酯萃取混合物，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，粗产物通过快速硅胶柱色谱(DCM:MeOH＝10:1)纯化，得到黄色固体状的所要目标产物41-1(680mg，85％产率)。DIEA (480 mg) was added to a solution of 1-4 (600 mg), tert-butyl 2,6-diazaspiro[3.3]heptane-2-carboxylate (245 mg) in dioxane (10 mL) at room temperature In, stirring atroom temperature 40 degrees. The mixture was poured into ice water, the mixture was extracted with ethyl acetate, the organic layer was washed with brine, dried overNa2SO4 and concentrated in vacuo, the crudeproduct was purified by flash silica gel column chromatography (DCM:MeOH=10:1) to give yellow The desired target product 41-1 (680 mg, 85% yield) as a solid.

之后参照实施例16的合成步骤完成目标产物41的合成。Then refer to the synthesis steps of Example 16 to complete the synthesis of target product 41.

实施例42、43和44的合成，参考41-1的合成分别用2,7-二氮杂螺[3.5]壬烷-7-甲酸叔丁酯、(S)-4-N-叔丁氧羰基-2-甲基哌嗪和叔丁基(S)-2-(氰基甲基)哌嗪-1-羧酸盐与共用中间体1-4反应得到相应的化合物，之后参考25的合成步骤完成目标产物42、43和44的合成。For the synthesis of Examples 42, 43 and 44, refer to the synthesis of 41-1 with 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester, (S)-4-N-tert-butoxy Carbonyl-2-methylpiperazine and tert-butyl (S)-2-(cyanomethyl)piperazine-1-carboxylate were reacted with common intermediates 1-4 to give the corresponding compounds, followed by the synthesis ofreference 25 The steps complete the synthesis of target products 42, 43 and 44.

Figure PCTCN2021124884-APPB-000082
Figure PCTCN2021124884-APPB-000082

Figure PCTCN2021124884-APPB-000083
Figure PCTCN2021124884-APPB-000083

实施例45：化合物45(1-(7-(7-(5-甲基-1H-吲唑-4-基)-2-(1-(2-(甲基磺酰基)乙基)哌啶-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 45: Compound 45 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(1-(2-(methylsulfonyl)ethyl)piperidine -4-yl)-8-(2,2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2 Synthesis of -yl)prop-2-en-1-one)

Figure PCTCN2021124884-APPB-000084
Figure PCTCN2021124884-APPB-000084

步骤1:化合物45-1的合成Step 1: Synthesis of compound 45-1

室温下，将化合物26-8(340mg)溶于THF(5mL)，加入TBAF(2mL，1M in THF)，35度搅拌过夜，浓缩，乙酸乙酯稀释，水洗2次，饱和食盐水洗涤两次，硫酸钠干燥，浓缩得产品(311mg，粗产品)。Dissolve compound 26-8 (340mg) in THF (5mL) at room temperature, add TBAF (2mL, 1M in THF), stir overnight at 35°C, concentrate, dilute with ethyl acetate, wash twice with water and twice with saturated brine , dried over sodium sulfate, and concentrated to give the product (311 mg, crude product).

步骤2：化合物45-2的合成Step 2: Synthesis of compound 45-2

室温下将化合物45-1(50mg)溶于乙醇(3mL)，加入甲基乙烯砜(68mg)，室温搅拌过夜，LCMS监控反应完毕，浓缩，乙酸乙酯稀释，水洗一次，饱和食盐水洗涤两次，干燥，浓缩得粗品，制备板(DCM/MeOH＝20:1)制备得45-2(32mg，36％收率)。ESI-MS m/z:882.46[M+H]+Dissolve compound 45-1 (50 mg) in ethanol (3 mL) at room temperature, add methyl vinyl sulfone (68 mg), stir overnight at room temperature, monitor the completion of the reaction by LCMS, concentrate, dilute with ethyl acetate, wash once with water, and wash twice with saturated saline. times, dried, and concentrated to obtain a crude product, and a preparative plate (DCM/MeOH=20:1) was prepared to obtain 45-2 (32 mg, 36% yield). ESI-MS m/z:882.46[M+H]+

步骤3：化合物45-3的合成Step 3: Synthesis of compound 45-3

在室温下，将45-2(32mg)溶于DCM(2mL)和TFA(1mL)的混合溶剂中，移至40℃反应约1h。待反应完全后，直接浓缩除去溶剂，得粗品目标产物45-3。ESI-MS m/z:698.40[M+H]+。45-2 (32 mg) was dissolved in a mixed solvent of DCM (2 mL) and TFA (1 mL) at room temperature, and moved to 40° C. for about 1 h. After the reaction was complete, the solvent was directly concentrated and removed to obtain the crude target product 45-3. ESI-MS m/z: 698.40 [M+H]+.

步骤4：化合物45的合成Step 4: Synthesis ofCompound 45

在氮气保护下，冰水浴中，将丙烯酰氯(4mg)的THF溶液加入到45-3(25mg粗品)的 THF(2mL)和饱和Na2CO3(0.5mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩物通过pre-TLC(DCM/MeOH＝10:1)纯化,得到所要产物45(6.6mg，24％产率)。ESI-MS m/z:752.45[M+H]+。Under the protection of nitrogen, the THF solution of acryloyl chloride (4 mg) was added to the THF (2 mL) and saturated Na2 CO3 (0.5 mL) solution of 45-3 (25 mg crude product) in an ice-water bath, and the reaction was complete upon addition. The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo, the concentrate was purified by pre-TLC (DCM/MeOH=10:1) to give the desired product 45 (6.6 mg, 24% yield). ESI-MS m/z: 752.45 [M+H]+.

实施例51：化合物51(1-(7-(7-(5-甲基-1H-吲唑-4-基)-2-(哌啶-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-yl)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 51: Compound 51 (1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(piperidin-4-yl)-8-(2,2,2 -trifluoroethoxy)-6-vinylquinazoline-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)prop-2-en-1-one) synthesis

Figure PCTCN2021124884-APPB-000085
Figure PCTCN2021124884-APPB-000085

步骤1:化合物51-1的合成Step 1: Synthesis of compound 51-1

将化合物26-8(100mg)溶于二氯甲烷(2mL)，加入盐酸二氧六环溶液(0.5mL，4N in dioxane)，反应液浓缩得粗品(50mg)，直接用于下一步。Compound 26-8 (100 mg) was dissolved in dichloromethane (2 mL), and dioxane hydrochloride solution (0.5 mL, 4N in dioxane) was added, and the reaction solution was concentrated to obtain a crude product (50 mg), which was directly used in the next step.

步骤2：化合物51-2的合成Step 2: Synthesis of Compound 51-2

在氮气保护下，冰水浴中，将丙烯酰氯(6mg)的THF溶液加入到51-1(50mg，粗品)的THF(2mL)和饱和Na2CO3(1mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩得粗品(30mg)。Under the protection of nitrogen, the THF solution of acryloyl chloride (6 mg) was added to the THF (2 mL) and saturated Na2 CO3 (1 mL) solution of 51-1 (50 mg, crude product) in an ice-water bath, and the reaction was complete upon addition. The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to afford crude product (30 mg).

步骤3：化合物51的合成Step 3: Synthesis of Compound 51

将化合物51-2(30mg)溶于二氯甲烷(1mL)，加入TFA(1mL)，室温搅拌2小时，反应完毕，浓缩，制备板制备(DCM/MeOH＝10：1)得目标产物(2.6mg，11％收率)。Compound 51-2 (30mg) was dissolved in dichloromethane (1mL), TFA (1mL) was added, stirred at room temperature for 2 hours, the reaction was completed, concentrated, and the preparation plate was prepared (DCM/MeOH=10:1) to obtain the target product (2.6 mg, 11% yield).

实施例47参考实施例1的步骤进行合成，用丙烯腈与共用中间体45-1反应即可得相应的目标产物，然后进行脱保护和引入丙烯酰基最终得到目标化合物47。Example 47 was synthesized with reference to the steps of Example 1, using acrylonitrile to react with common intermediate 45-1 to obtain the corresponding target product, and then deprotected and introduced acryloyl group to finally obtain the target compound 47.

实施例46，53，54，55，56，57，59，60，61，62，65，66，67，68，70和71参考实施例28采用还原胺化的方法，用不同的酮或者醛与共用中间体45-1反应得到相应的产物，之后再脱保护和引入乙酰基合成最终目标产物；实施例48参考实施例27，用共用中间体45-1与甲基环氧丙烷反应生成对应的产物，然后脱保护和引入丙烯酰基最终得到目标化合物48；实施例49，58，63，64，69采用烷基化的策略与共用中间体45-1反应生成对应的产物，然后脱保护和引入丙烯酰基最终得到最终目标产物。对应的合成中间体如下表:Embodiment 46, 53, 54, 55, 56, 57, 59, 60, 61, 62, 65, 66, 67, 68, 70 and 71 Reference Example 28 adopts the method of reductive amination, with different ketones or aldehydes React with common intermediate 45-1 to obtain the corresponding product, and then deprotect and introduce acetyl to synthesize the final target product; Example 48 refers to Example 27, using common intermediate 45-1 to react with methyl propylene oxide to generate the corresponding Then deprotect and introduce acryloyl to finally obtain the target compound 48; Examples 49, 58, 63, 64, and 69 use the strategy of alkylation to react with the common intermediate 45-1 to generate the corresponding product, then deprotect and The introduction of acryloyl group finally obtains the final target product. The corresponding synthetic intermediates are as follows:

Figure PCTCN2021124884-APPB-000086
Figure PCTCN2021124884-APPB-000086

Figure PCTCN2021124884-APPB-000087
Figure PCTCN2021124884-APPB-000087

Figure PCTCN2021124884-APPB-000088
Figure PCTCN2021124884-APPB-000088

Figure PCTCN2021124884-APPB-000089
Figure PCTCN2021124884-APPB-000089

Figure PCTCN2021124884-APPB-000090
Figure PCTCN2021124884-APPB-000090

Figure PCTCN2021124884-APPB-000091
Figure PCTCN2021124884-APPB-000091

Figure PCTCN2021124884-APPB-000092
Figure PCTCN2021124884-APPB-000092

Figure PCTCN2021124884-APPB-000093
Figure PCTCN2021124884-APPB-000093

Figure PCTCN2021124884-APPB-000094
Figure PCTCN2021124884-APPB-000094

Figure PCTCN2021124884-APPB-000095
Figure PCTCN2021124884-APPB-000095

Figure PCTCN2021124884-APPB-000096
Figure PCTCN2021124884-APPB-000096

Figure PCTCN2021124884-APPB-000097
Figure PCTCN2021124884-APPB-000097

实施例72：化合物72(2-氟-1-(7-(7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-8-(2,2,2-三氟乙氧基)-6-乙烯基喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)丙-2-烯-1-酮)的合成Example 72: Compound 72 (2-fluoro-1-(7-(7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidin-4-yl)- 8-(2,2,2-trifluoroethoxy)-6-vinylquinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)propan-2 -en-1-one) synthesis

Figure PCTCN2021124884-APPB-000098
Figure PCTCN2021124884-APPB-000098

将化合物2-8(120mg)溶于DMF(5mL)，加入2-氟丙烯酸(54mg)，HATU(226mmol)，DIEA(77mg)，搅拌2小时，反应完毕，乙酸乙酯稀释，饱和食盐水洗涤三次，干燥浓缩得粗品，制备板制备(DCM/MeOH＝10/1)得目标产物(3mg)。Compound 2-8 (120 mg) was dissolved in DMF (5 mL), 2-fluoroacrylic acid (54 mg), HATU (226 mmol), DIEA (77 mg) was added, stirred for 2 hours, the reaction was completed, diluted with ethyl acetate, washed with saturated brine Three times, dried and concentrated to get the crude product, the preparative plate was prepared (DCM/MeOH=10/1) to get the target product (3 mg).

实施例75:化合物75((E)-1-(7-(6-氯-7-(5-甲基-1H-吲唑-4-基)-2-(1-甲基哌啶-4-基)-8-(2,2,2-三氟乙氧基)喹唑啉-4-基)-2,7-二氮杂螺[3.5]壬基-2-基)-4-(二甲基氨基) 丁-2-烯-1-酮)的合成Example 75: Compound 75 ((E)-1-(7-(6-chloro-7-(5-methyl-1H-indazol-4-yl)-2-(1-methylpiperidine-4 -yl)-8-(2,2,2-trifluoroethoxy)quinazolin-4-yl)-2,7-diazaspiro[3.5]nonyl-2-yl)-4-( Synthesis of dimethylamino)but-2-en-1-one)

Figure PCTCN2021124884-APPB-000099
Figure PCTCN2021124884-APPB-000099

在氮气保护下，冰水浴中，将新鲜制备的反式-4-二甲基胺基巴豆酰氯盐酸盐(12mg)的THF溶液加入到23-6(100mg，粗品)的THF(2mL)和饱和Na2CO3(1mL)溶液中，加入即反应完全。将反应混合物倒入水中，用乙酸乙酯萃取，有机层用盐水洗涤，经Na2SO4干燥且真空浓缩，浓缩得粗品，粗品制备板制备(DCM/MeOH＝10：1)(6mg，5％收率).Under nitrogen protection, in an ice-water bath, a THF solution of freshly prepared trans-4-dimethylaminocrotonyl chloride hydrochloride (12 mg) was added to THF (2 mL) of 23-6 (100 mg, crude product) and Saturated Na2 CO3 (1 mL) solution, the reaction is complete upon addition. The reaction mixture was poured into water, extracted with ethyl acetate, the organic layer was washed with brine, dried over Na2 SO4 and concentrated in vacuo to give a crude product, which was prepared in a plate (DCM/MeOH=10:1) (6 mg, 5 % yield).

实施例73合成参考50的合成步骤，不同点是用二氟乙醇替代三氟乙醇。The synthesis of Example 73 refers to the synthesis steps of 50, the difference is that trifluoroethanol is replaced by difluoroethanol.

实施例74合成参考72的合成步骤。Synthesis of Example 74 Synthesis of Reference 72.

Figure PCTCN2021124884-APPB-000100
Figure PCTCN2021124884-APPB-000100

Figure PCTCN2021124884-APPB-000101
Figure PCTCN2021124884-APPB-000101

参照实施例1A，采用基本相同的方法对实施例2～75化合物进行了拆分，分别获得了对应的手性异构体2a～75a。例如，化合物9a结构式为

Figure PCTCN2021124884-APPB-000102
化合物47a结构式为
Figure PCTCN2021124884-APPB-000103
Referring to Example 1A, the compounds of Examples 2-75 were resolved by basically the same method, and the corresponding chiral isomers 2a-75a were respectively obtained. For example, the structural formula of compound 9a is
Figure PCTCN2021124884-APPB-000102
The structural formula of compound 47a is
Figure PCTCN2021124884-APPB-000103

对比化合物1(D1)的合成Synthesis of comparative compound 1 (D1)

AMG510(D1)合成参考WO2018217651A1。The synthesis of AMG510(D1) refers to WO2018217651A1.

Figure PCTCN2021124884-APPB-000104
Figure PCTCN2021124884-APPB-000104

对比化合物2(D2)的合成Synthesis of comparative compound 2 (D2)

D2合成参考WO2018143315A1。D2 synthesis refers to WO2018143315A1.

Figure PCTCN2021124884-APPB-000105
Figure PCTCN2021124884-APPB-000105

药理实验Pharmacological experiment

实施例A：细胞增殖抑制实验Embodiment A: Cell Proliferation Inhibition Experiment

将CALU-1细胞按1000细胞、190μL/孔铺96孔超低吸附板。孵育隔夜后，配制梯度浓度的化合物溶液，分别向各孔细胞中加入10μL各浓度的待测化合物DMSO溶液，化合物终浓度为30000、10000、3333.3、1111.1、370.4、123.5、41.2、13.7、4.6、0nM(DMSO终浓度均为0.25％)。37℃，5％CO2孵育120小时。向各孔中加入60μL Cell-titer Glo工作液，震荡混匀后室温孵育10分钟，多功能酶标仪读取Luminescence发光值，将发光值读数转换为抑制百分数：Spread 1000 cells of CALU-1 cells, 190 μL/well on a 96-well ultra-low adsorption plate. After incubating overnight, compound solutions with gradient concentrations were prepared, and 10 μL of DMSO solutions of the compounds to be tested were added to the cells in each well. 0 nM (the final concentration of DMSO is 0.25%). Incubate for 120 hours at 37 °C, 5%CO2 . Add 60 μL of Cell-titer Glo working solution to each well, shake and mix well, and incubate at room temperature for 10 minutes. Read the luminescence value of Luminescence with a multi-functional microplate reader, and convert the luminescence value reading into the percentage of inhibition:

抑制百分数＝(最大值-读数)/(最大值-最小值)*100。Percent Inhibition=(Maximum-Reading)/(Maximum-Minimum)*100.

“最大值”为DMSO对照；“最小值”表示无细胞对照组。"Maximum" is the DMSO control; "minimum" is the no-cell control.

用Graphpad Prism软件进行曲线拟合并得到IC50值。Curve fitting was performed with Graphpad Prism software andIC50 values were obtained.

实施例化合物对CALU-1细胞具有良好的抑制活性，特别是Y取代基选自

Figure PCTCN2021124884-APPB-000106
Figure PCTCN2021124884-APPB-000107
时，相对于其他通过环上C原子与喹唑啉环直连相连的环结构分子具有更优活性。 实施例合物对CALU-1细胞抑制的IC50数据参见表2。Example compounds have good inhibitory activity on CALU-1 cells, especially the Y substituent is selected from
Figure PCTCN2021124884-APPB-000106
Figure PCTCN2021124884-APPB-000107
, it has better activity than other ring structure molecules that are directly connected to the quinazoline ring through the C atom on the ring. See Table 2 for the IC50 data of the compound of the example on the inhibition of CALU-1 cells.

表2Table 2

化合物名称Compound nameIC50(nM)IC50 (nM)化合物1Compound 12.72.7化合物2Compound 2776.1776.1化合物3Compound 3277.6277.6化合物4Compound 42.92.9化合物5Compound 545.745.7化合物6Compound 64.64.6

实施例B：细胞增殖抑制实验Example B: Cell Proliferation Inhibition Experiment

将MIA-PACA-2细胞按600细胞、160μL/孔铺96孔超低吸附板。孵育隔夜后，配制梯度浓度的化合物溶液，分别向各孔细胞中加入40μL各浓度的待测化合物DMSO溶液，化合物终浓度为10000、2000、400、80、16、3.2、0.64、0.12、0.025、0nM(DMSO终浓度均为0.25％)。37℃，5％CO2孵育96h。向各孔中加入50μL Cell-titer Glo工作液，震荡混匀后室温孵育10min，多功能酶标仪读取Luminescence发光值，将发光值读数转换为抑制百分数：MIA-PACA-2 cells were plated on 96-well ultra-low adsorption plates at 600 cells, 160 μL/well. After incubating overnight, compound solutions with gradient concentrations were prepared, and 40 μL of each concentration of the compound to be tested in DMSO was added to the cells in each well. 0 nM (the final concentration of DMSO is 0.25%). Incubate at 37°C, 5% CO2 for 96h. Add 50 μL of Cell-titer Glo working solution to each well, shake and mix well, and incubate at room temperature for 10 minutes. Read the luminescence value of Luminescence with a multi-functional microplate reader, and convert the luminescence value reading into the percentage of inhibition:

抑制百分数＝(最大值-读数)/(最大值-最小值)*100。Percent Inhibition=(Maximum-Reading)/(Maximum-Minimum)*100.

“最大值”为DMSO对照；“最小值”表示无细胞对照组。"Maximum" is the DMSO control; "minimum" is the no-cell control.

用Graphpad Prism软件进行曲线拟合并得到IC50值。Curve fitting was performed with Graphpad Prism software andIC50 values were obtained.

实施例部分化合物对MIA-PACA-2细胞抑制的IC50数据参见表3。See Table 3 for the IC50 data of the compounds in the examples for inhibiting MIA-PACA-2 cells.

实施例C：细胞增殖抑制实验Example C: Cell Proliferation Inhibition Experiment

将H358细胞按2000细胞、190μL/孔铺96孔超低吸附板。孵育隔夜后，配制梯度浓度的化合物溶液，分别向各孔细胞中加入10μL各浓度的待测化合物DMSO溶液，化合物终浓度为10000、3333.3、1111.1、370.4、123.5、41.2、13.7、4.6、1.5、0nM(DMSO终浓度均为0.25％)。37℃，5％CO2孵育96h。向各孔中加入50μL Cell-titer Glo工作液，震荡混匀后室温孵育10min，多功能酶标仪读取Luminescence发光值，将发光值读数转换为抑制百分数：The H358 cells were plated in 96-well ultra-low adsorption plates as 2000 cells, 190 μL/well. After incubation overnight, compound solutions with gradient concentrations were prepared, and 10 μL of DMSO solutions of the compounds to be tested were added to the cells in each well. 0 nM (the final concentration of DMSO is 0.25%). Incubate at 37°C, 5% CO2 for 96h. Add 50 μL of Cell-titer Glo working solution to each well, shake and mix well, and incubate at room temperature for 10 minutes. Read the luminescence value of Luminescence with a multi-functional microplate reader, and convert the luminescence value reading into the percentage of inhibition:

抑制百分数＝(最大值-读数)/(最大值-最小值)*100。Percent Inhibition=(Maximum-Reading)/(Maximum-Minimum)*100.

“最大值”为DMSO对照；“最小值”表示无细胞对照组。 用Graphpad Prism软件进行曲线拟合并得到IC50值。"Maximum" is the DMSO control; "minimum" is the no-cell control. Curve fitting was performed with Graphpad Prism software andIC50 values were obtained.

实施例部分化合物对H358细胞抑制的IC50数据参见表3。See Table 3 for the IC50 data of the compounds in the examples for inhibition of H358 cells.

表3table 3

Figure PCTCN2021124884-APPB-000108
Figure PCTCN2021124884-APPB-000108

Figure PCTCN2021124884-APPB-000109
Figure PCTCN2021124884-APPB-000109

实施例D：异构体活性Example D: Isomer Activity

使用色谱条件：CHIRALCEL OD(2.5cm I.D.×25cm L,10μm)，UV 254nm，流动相Hexane/IPA/DEA＝70/30/0.1(V/V/V)，流速20ml/min,将化合物进行拆分，然后采用实施例B和实施例C方法测定异构体活性。Chromatographic conditions used: CHIRALCEL OD (2.5cm I.D.×25cm L, 10μm), UV 254nm, mobile phase Hexane/IPA/DEA=70/30/0.1 (V/V/V), flow rate 20ml/min, the compound was resolved Point, then adopt embodiment B and embodiment C method to measure isomer activity.

结果表明，本发明通式(I)所示手性化合物的代表性异构体活性优于其对应轴手性异构体，活性差异大于100倍。The results show that the activity of the representative isomer of the chiral compound represented by the general formula (I) of the present invention is better than that of the corresponding axial chiral isomer, and the activity difference is greater than 100 times.

表4Table 4

Figure PCTCN2021124884-APPB-000110
Figure PCTCN2021124884-APPB-000110

表5table 5

Figure PCTCN2021124884-APPB-000111
Figure PCTCN2021124884-APPB-000111

实施例E：人胰腺癌MIA PaCa-2 CDX肿瘤模型上的体内药效学研究Example E: In vivo pharmacodynamic study on human pancreatic cancer MIA PaCa-2 CDX tumor model

用经典的小鼠肿瘤模型试验，观察灌胃给药后，目标化合物抑瘤率TGI(％)，以评价其抗肿瘤活性。The classic mouse tumor model test is used to observe the tumor inhibition rate TGI (%) of the target compound after intragastric administration, so as to evaluate its anti-tumor activity.

实验方法：人胰腺癌MIA PaCa-2细胞体外单层培养，培养条件为DMEM/F12培养基中加10％胎牛血清，1％双抗，37℃5％CO2孵箱培养。一周两次进行常规处理传代。当细胞饱和度为80％-90％，数量到达要求时，收取细胞，计数，接种。将0.2mL(5×106个)MIA PaCa-2细胞(加基质胶，体积比为1:1)皮下接种于每只小鼠的右后背，肿瘤平均体积达到约149mm3时开始分组给药。Experimental method: Human pancreatic cancer MIA PaCa-2 cells were cultured in vitro as a monolayer. The culture conditions were DMEM/F12 medium plus 10% fetal bovine serum, 1% double antibody, and cultured in a 5% CO2 incubator at 37°C. Conventional passaging was performed twice a week. When the cell saturation is 80%-90% and the number reaches the requirement, the cells are collected, counted, and inoculated. 0.2mL (5×106 ) MIA PaCa-2 cells (plus Matrigel, volume ratio 1:1) were inoculated subcutaneously on the right back of each mouse, and the tumors were divided into groups when the average tumor volume reached about149mm3 medicine.

实验结果：目标化合物1a和AMG510均可有效降低肿瘤TGI，与溶剂对照组有显著差异(图4)。目标化合物1a处理小鼠体重变化情况更优，显示出更好安全性潜力(图5)。Experimental results: Both thetarget compound 1a and AMG510 can effectively reduce tumor TGI, which is significantly different from the solvent control group (Figure 4). The body weight change of the mice treated with thetarget compound 1a is better, showing better safety potential (Figure 5).

实施例F：人非小细胞肺癌PDX肿瘤模型的体内药效学研究Example F: In vivo pharmacodynamic study of human non-small cell lung cancer PDX tumor model

用经典的小鼠肿瘤模型试验，观察灌胃给药后，目标化合物抑瘤率TGI(％)，以评价其抗肿瘤活性。The classic mouse tumor model test is used to observe the tumor inhibition rate TGI (%) of the target compound after intragastric administration, so as to evaluate its anti-tumor activity.

实验方法：人源肺癌LU-01-0030PDX模型的建立最初来源于外科手术切除的G12C突变阳性患者临床样本，在植入裸鼠体内后被定义为P0代。将P0代的肿瘤组织植入下一代被称为P1代。以此类推持续在裸鼠体内植入。其中FP3的肿瘤是通过P2代重新复苏得到的。由FP3代产生的下一代被定为FP4，以此类推。FP5代的肿瘤组织皮下接种于每只小鼠的右上肢，肿瘤平均体积达到约141mm3时开始分组给药。Experimental method: The establishment of the human-derived lung cancer LU-01-0030PDX model was initially derived from surgically resected clinical samples of G12C mutation-positive patients, which were defined as the P0 generation after implantation into nude mice. Implantation of tumor tissue from generation P0 into the next generation is referred to as generation P1. Continue to implant in nude mice by analogy. Among them, FP3 tumors were resuscitated from the P2 generation. The next generation generated by the FP3 generation is designated as FP4, and so on. The tumor tissue of passage FP5 was inoculated subcutaneously on the right upper limb of each mouse, and the group administration began when the average tumor volume reached about 141mm3.

实验结果：人非小细胞肺癌LU-01-0030PDX模型上，给予10mg/kg AMG 510观察到微弱抑瘤效果，TGI为3.3％；给予3mg/kg、10mg/kg或30mg/kg的目标化合物1a,TGI％分别为32％、28％和52％，效果优于AMG 510(图6)。化合物1a显示出良好耐药性。Experimental results: On the LU-01-0030PDX model of human non-small cell lung cancer, a weak tumor inhibitory effect was observed after administration of 10mg/kg AMG 510, with a TGI of 3.3%; administration of 3mg/kg, 10mg/kg or 30mg/kg of thetarget compound 1a , TGI% were 32%, 28% and 52%, the effect is better than AMG 510 (Figure 6).Compound 1a showed good drug resistance.

实施例G：hERG测试Example G: hERG test

1稳转细胞准备1 Stably transfected cell preparation

细胞株来源于过表达hERG钾离子通道HEK293细胞，细胞在37℃、5％CO2培养箱中培养。当细胞密度达培养皿80％时，先用磷酸盐缓冲液(PBS)预清洗，然后用胰蛋白酶/EDTA消化细胞2-3分钟，加入细胞培养基停止消化，轻轻用移液管吹打并转移到离心管中，1000转/分钟离心3分钟，上清液倒掉，加入细胞培养基，轻轻吹打将细胞混匀，随后转移到培养皿中进行传代培养，或将细胞滴于圆形玻片上在细胞培养液中培养待细胞贴壁用于实验。The cell line is derived from HEK293 cells overexpressing the hERG potassium ion channel, and the cells are cultured in a 37°C, 5% CO2 incubator. When the cell density reaches 80% of the culture dish, pre-wash with phosphate buffered saline (PBS), then digest the cells with trypsin/EDTA for 2-3 minutes, add cell culture medium to stop the digestion, gently pipette and remove Transfer to a centrifuge tube, centrifuge at 1000 rpm for 3 minutes, pour off the supernatant, add cell culture medium, gently pipette to mix the cells, then transfer to a petri dish for subculture, or drop the cells on a circular Cells were cultured in cell culture medium on slides until they adhered to the wall for experiments.

细胞培养基组成：DMEM、15％胎牛血清和1％100x青霉素-链霉素。Cell culture medium composition: DMEM, 15% fetal calf serum and 1% 100x penicillin-streptomycin.

2溶液配制2 solution preparation

细胞内液和外液的组成成分见表4。The composition of intracellular fluid and extracellular fluid is shown in Table 4.

表6Table 6

Figure PCTCN2021124884-APPB-000112
Figure PCTCN2021124884-APPB-000112

3电生理手动膜片钳系统实验方案3 Experimental scheme of electrophysiological manual patch clamp system

将稳转的细胞滴于圆形玻片上并置于培养皿中，细胞密度低于50％，培养过夜。将实验用细胞转移到一个嵌于倒置显微镜平台的细胞浴槽中，灌流细胞外液，灌流速度为2.7ml/ 分钟。稳定5分钟后即可开始实验。采用HEKA EPC-10膜片钳放大器和PATCHMASTER采集系统记录膜电流(HEKA Instruments Inc.,D-67466Lambrecht，Pfalz，Germany)。所有实验均在室温(22-24℃)下完成。实验中使用P-97微电极拉制仪(Sutter Instrument Company，One Digital Drive，Novato，CA 94949)拉直电极(BF150-110-10)。电极内径为1-1.5mm，充满内液后的入水电阻为2-4MΩ。The stably transfected cells were dropped on a round glass slide and placed in a petri dish at a cell density below 50%, and cultured overnight. The experimental cells were transferred to a cell bath embedded in an inverted microscope platform, and the extracellular fluid was perfused at a rate of 2.7ml/min. The experiment can be started after 5 minutes of stabilization. Membrane currents were recorded using a HEKA EPC-10 patch clamp amplifier and PATCHMASTER acquisition system (HEKA Instruments Inc., D-67466 Lambrecht, Pfalz, Germany). All experiments were done at room temperature (22-24°C). The electrode (BF150-110-10) was straightened using a P-97 microelectrode puller (Sutter Instrument Company, One Digital Drive, Novato, CA 94949) in the experiment. The inner diameter of the electrode is 1-1.5mm, and the water resistance after being filled with inner liquid is 2-4MΩ.

hERG钾通道的电生理刺激方案，是首先将膜电压钳制在-80mV，给予细胞持续2s,+20mV电压刺激，激活hERG钾通道，再复极化至-50mV,持续5s，产生外向尾电流，刺激频率每15s一次。电流值为尾电流的峰值。The electrophysiological stimulation scheme of the hERG potassium channel is to first clamp the membrane voltage at -80mV, give the cells a continuous 2s, +20mV voltage stimulation, activate the hERG potassium channel, and then repolarize to -50mV for 5s to generate an outward tail current. The stimulation frequency was once every 15s. The current value is the peak value of the tail current.

实验中采用全细胞记录模式记录通道电流。首先灌流细胞外液(大约每分钟2毫升)并持续记录，并等待电流稳定(5分钟内电流衰减(Run-Down)小于5％)，此时尾电流峰值即为对照电流值。接着灌流含待测药物的细胞外液并持续记录直到药物对hERG电流的抑制作用到达稳定状态，此时尾电流峰值即为加药后电流值。稳定状态的标准以最近的连续3个电流记录线是否重合来判断。达到稳定态势以后如果以细胞外液灌流冲洗后hERG电流回复或接近加药物之前的大小，则可以继续灌流测试其它浓度或药物。30μM Quinidine(奎尼丁)被用于实验中作为阳性对照以保证所使用的细胞反应正常。In the experiment, the channel current was recorded in the whole-cell recording mode. First, perfuse the extracellular fluid (about 2 ml per minute) and continue to record, and wait for the current to stabilize (the current decay (Run-Down) is less than 5% within 5 minutes), at this time the peak value of the tail current is the control current value. Then perfuse the extracellular fluid containing the drug to be tested and keep recording until the inhibitory effect of the drug on the hERG current reaches a steady state. At this time, the peak value of the tail current is the current value after adding the drug. The standard of steady state is judged by whether the latest three continuous current recording lines coincide. After reaching a steady state, if the hERG current returns to or approaches the value before adding drugs after perfusion and flushing with extracellular fluid, you can continue to test other concentrations or drugs by perfusion. 30 μM Quinidine (quinidine) was used in the experiment as a positive control to ensure that the cells used responded normally.

4化合物准备4 Compound preparation

用DMSO稀释母液至下列浓度(次级母液)，分别为：3.3、1.1、0.37、0.12mM，取母液和次级母液各30μl稀释至10ml细胞外液中，用于电生理检测；Dilute the mother solution with DMSO to the following concentrations (secondary mother solution), respectively: 3.3, 1.1, 0.37, and 0.12 mM, take 30 μl of each of the mother solution and the secondary mother solution and dilute it into 10 ml of extracellular fluid for electrophysiological testing;

化合物的终浓度为30,10,3.3,1.1,0.37μM；The final concentration of the compound is 30, 10, 3.3, 1.1, 0.37 μM;

DMSO的终浓度为3:1000。The final concentration of DMSO was 3:1000.

5数据分析5Data analysis

实验数据使用PATCHMASTER V2X60(HEKA Instruments Inc.,D-67466 Lambrecht，Pfalz，Germany)采集，并采用Origin 8.5(OriginLab Corporation，Northampton，MA)软件以及Microsoft Excel进行分析和统计。The experimental data were collected using PATCHMASTER V2X60 (HEKA Instruments Inc., D-67466 Lambrecht, Pfalz, Germany), and were analyzed and counted using Origin 8.5 (OriginLab Corporation, Northampton, MA) software and Microsoft Excel.

通过测量对照组与药物处理组的电流最大值，计算处理组最大电流值所占对照组最大电流值的比率，评估待测化合物在测试浓度下对hERG钾离子通道的作用效果(Mean±SE)。By measuring the maximum current value of the control group and the drug treatment group, calculate the ratio of the maximum current value of the treatment group to the maximum current value of the control group, and evaluate the effect of the test compound on the hERG potassium ion channel at the test concentration (Mean±SE) .

6质量控制6 Quality Control

本研究按照科瑞斯实验室标准操作规程(近似GLP规范)执行，同时所有实验方法均参考同行评议期刊发表的文献，并按照科瑞斯生物的标准实验操作规程执行。This study was carried out in accordance with the standard operating procedures of Keruisi Laboratory (approximately GLP specifications). At the same time, all experimental methods refer to the literature published in peer-reviewed journals, and are carried out in accordance with the standard operating procedures of Keruisi Biology.

报告中的试验数据需要满足以下标准：The test data in the report needs to meet the following criteria:

电生理记录参数Electrophysiological recording parameters

封接电阻>500MΩSeal resistance>500MΩ

直联电阻(Rs)<10MΩDirect resistance (Rs)<10MΩ

初始尾电流幅度>300pAInitial tail current amplitude >300pA

电流rundown(自发性减小)<2％/每分钟Current rundown (spontaneous decrease) <2%/min

漏电流<200pA或者hERG电流峰值的10％(在90％的记录时间之内)Leakage current <200pA or 10% of peak hERG current (during 90% of recording time)

7实验结果：7 Experimental results:

表7Table 7

化合物compoundhERG IC50(uM)hERG IC50 (uM)1a1a>30>30D2D28.128.12

8实验结论：8 Experimental conclusions:

药物对于心脏hERG钾离子通道的抑制是药物导致QT延长综合症的主要原因。化合物1a在测试浓度范围内对hERG钾通道几乎无阻滞作用(IC50>30μM)；化合物D2在测试浓度范围内对hERG钾通道有中度的阻滞作用(3μM<预测IC50<10μM)，化合物存在一定程度的因为对hERG的作用而导致的心脏风险。Inhibition of cardiac hERG potassium channels by drugs is the main cause of long QT syndrome caused by drugs.Compound 1a has almost no blocking effect on hERG potassium channel in the tested concentration range (IC50 >30μM); compound D2 has moderate blocking effect on hERG potassium channel in the tested concentration range (3μM<predicted IC50 <10μM) , the compound presents some degree of cardiac risk due to its effect on hERG.

实施例H：体外CYP酶抑制测试Example H: In vitro CYP enzyme inhibition test

采用人肝微粒体孵化体系，通过代谢物的生成量来反映各种酶的活性。The human liver microsome incubation system is used to reflect the activities of various enzymes through the generation of metabolites.

实验方法：称取适量的待测物标准品，用DMSO稀释至2mM，得到工作液；探针底物用纯乙腈配制，阳性对照抑制剂用DMSO配制，浓度详见表8。取待测化合物/阳性对照抑制剂1μL、探针底物1μL、人肝微粒体(20mg/mL)1μL和PBS缓冲液177μL制备反应混合物(孵化反应待测化合物/阳性对照抑制剂终浓度为10μM)。上述混合物在37℃预孵育5分钟。向反应混合物加入20μL NADPH溶液(10mM，PBS配制)，在37℃条件下CYP2C8、CYP2C9酶孵化10分钟，CYP2C19酶孵化30分钟。所有孵育样品设双样本。加入200μL冰冷含内标的乙腈终止反应。在3200rpm离心15分钟。上清液转移至LC-MS/MS分析。通过代谢物的生产量来反映各种酶的活性。采用单点法计算，公式如下：Experimental method: Weigh an appropriate amount of standard substance to be tested and dilute it to 2mM with DMSO to obtain a working solution; the probe substrate is prepared with pure acetonitrile, and the positive control inhibitor is prepared with DMSO. The concentrations are shown in Table 8. Take 1 μL of the test compound/positive control inhibitor, 1 μL of the probe substrate, 1 μL of human liver microsomes (20 mg/mL) and 177 μL of PBS buffer to prepare a reaction mixture (the final concentration of the test compound/positive control inhibitor in the incubation reaction is 10 μM ). The above mixture was pre-incubated at 37°C for 5 minutes. 20 μL of NADPH solution (10 mM, prepared in PBS) was added to the reaction mixture, and CYP2C8 and CYP2C9 enzymes were incubated at 37° C. for 10 minutes, and CYP2C19 enzymes were incubated for 30 minutes. All incubation samples were set as double samples. The reaction was terminated by adding 200 μL of ice-cold acetonitrile containing internal standard. Centrifuge at 3200 rpm for 15 minutes. The supernatant was transferred to LC-MS/MS analysis. The activity of various enzymes is reflected by the production of metabolites. Calculated using the single-point method, the formula is as follows:

Inhibiton％＝100％*(1-Test compound Aear ratio/Control Aear ratio)，实验结果见表9。Inhibiton%=100%*(1-Test compound Aear ratio/Control Aear ratio), the experimental results are shown in Table 9.

表8Table 8

代谢酶metabolic enzyme探针底物/浓度Probe substrate/concentration阳性对照/浓度Positive Control/ConcentrationCYP2C8CYP2C8Paclitaxel/2mMPaclitaxel/2mMQuercertin/2mMQuercertin/2mMCYP2C9CYP2C9Diclofenac/1mMDiclofenac/1mMSulfaphenazole/2mMSulfaphenazole/2mMCYP2C19CYP2C19S-Mephenytoin/10mMS-Mephenytoin/10mM(+)-N-3-Benzylnirvanol/2mM(+)-N-3-Benzylnirvanol/2mM

表9％Inhibition on major CYP isoforms@10μMTable 9% Inhibition on major CYP isoforms@10μM

Figure PCTCN2021124884-APPB-000113
Figure PCTCN2021124884-APPB-000113

实验结果：本发明的优选化合物对于CYP2C8、CYP2C9和CYP2C19酶具有较弱程度的抑制作用，因此在临床给药时，药物代谢性相互作用可能性较低。Experimental results: The preferred compounds of the present invention have a weaker inhibitory effect on CYP2C8, CYP2C9 and CYP2C19 enzymes, so the possibility of drug metabolic interaction is low during clinical administration.

虽然本发明已通过其实施方式进行了全面的描述，但是值得注意的是，各种变化和修改对于本领域技术人员都是显而易见的。这样的变化和修改都应该包括在本发明所附权利要求的范围内。While the invention has been fully described by way of the embodiments thereof, it is to be noted that various changes and modifications will be apparent to those skilled in the art. Such changes and modifications should all be included within the scope of the appended claims of the present invention.

Claims (15)

  1. A chiral compound shown in a general formula (I), deuterated compound or medicinal salt,
    Figure PCTCN2021124884-APPB-100001
    wherein X is selected from C3-14 Cycloalkyl, 3-14 membered heterocyclyl, 3-14 membered heterocyclylamino; x is optionally further substituted with 1-4 substituents, wherein each substituent is independently selected from C1-6 Alkyl, amino, C1-6 Aminoalkyl, carbamoyl, C1-6 Carbamoyl alkyl, carboxyl, C1-6 Carboxyalkyl, cyano, C1-6 Cyanoalkyl, halogen, C1-6 Haloalkyl, hydroxy, C1-6 A hydroxyalkyl group;
    R1 Is an acryl, substituted acryl or
    Figure PCTCN2021124884-APPB-100002
    R2 Selected from H, C2-6 Alkenyl, C1-6 Alkoxy, C1-6 Haloalkoxy, C1-6 Alkyl, 4-10 membered heterocyclyl substituted C1-6 Alkyl, C1-6 Alkylsulfonyl, C1-6 Alkylsulfinyl, C1-6 Alkylthio, C2-6 Alkynyl, C1-6 Alkylamino, amino, C1-6 Aminoalkyl, carbamoyl, C1-6 Carbamoyl alkyl, C1-6 Carboxyalkyl, cyano, C1-6 Cyanoalkyl, halogen, C1-6 Haloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted 5-10 membered heteroaryl, substituted or unsubstituted C3-7 Cycloalkyl, substituted or unsubstituted 4-10 membered heterocyclyl, hydroxy or oxo;
    R3 selected from H, halogen, C1-6 Alkyl, substituted C1-6 Alkyl, C2-6 Alkenyl, substituted C2-6 Alkenyl, C3-6 Cycloalkyl or substituted C3-6 Cycloalkyl;
    R4 or R is5 Independently selected from H, halogen, C1-6 Alkyl, substituted C1-6 Alkyl, C3-6 Cycloalkyl, substituted C3-6 Cycloalkyl;
    R6 selected from hydroxy, oxo, C1-6 Alkoxy, C3-10 Cycloalkyloxy-and cyano-substituted cyclopropyl C1-6 Alkylene oxide group, the C1-6 The alkoxy groups are optionally further substituted with one or more groups selected from halogen, hydroxy, C1-6 Alkoxy, C3-8 Substituted cycloalkyl;
    m or n is independently selected from 0, 1, 2, 3, 4;
    Y is selected from 3-14 membered heterocyclyl directly attached to the quinazoline ring through a C atom on the ring, said heterocyclyl optionally being further substituted with 1-4 substituents selected from R7 Or R is8 Is substituted by a substituent of (2); r is R7 Or R is8 Independently selected from H, =O, =S, cyano,Halogen, nitro, C1-6 Alkyl, -C0-6 alkylene-ORa 、-C0-6 alkylene-OC (O) N (R)a )2 、-C0-6 alkylene-N (R)a )2 、-C0-6 alkylene-NRa C(O)Ra 、-C0-6 alkylene-NRa C(O)N(Ra )2 、-C0-6 alkylene-NRa S(O)Ra 、-C0-6 alkylene-NRa S(O)2 Ra 、-C0-6 alkylene-S (=o) Ra 、-C0-6 alkylene-S (=o)2 Ra 、-C0-6 alkylene-SRa 、-C0-6 alkylene-S (R)a )5 、-C0-6 alkylene-C (=o) Ra 、-C0-6 alkylene-C (=o) ORa 、-C0-3 alkylene-C (=O) N (R)a )2 、C2-6 Alkenyl, C2-6 Alkynyl, acryl, -C0-6 alkylene-C3-14 Cycloalkyl, -C0-6 Alkylene- (3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 Aryl or-C0-6 Alkylene- (5-14 membered heteroaryl), said C1-6 Alkyl, C2-6 Alkenyl, C2-6 Alkynyl, -C0-6 alkylene-C3-14 Cycloalkyl, -C0-6 Alkylene- (3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 Aryl or-C0-6 Alkylene- (5-14 membered heteroaryl) optionally may also be substituted with 1 or more Ra Substituted; each Ra Each independently selected from H, halogen, hydroxy, amino, oxo, nitro, cyano, carboxyl, C1-6 Alkyl group、C1-6 Hydroxyalkyl, C1-6 Aminoalkyl, C1-6 Haloalkyl, C1-6 Alkoxy, C1-6 Haloalkoxy, C1-6 Heteroalkyl, C3-8 Cycloalkyl, 3-8 membered heterocyclyl, C6-14 Aryl, 5-14 membered heteroaryl, C1-6 Acyl or
    Figure PCTCN2021124884-APPB-100003
  2. Chiral compound, deuterated or pharmaceutically acceptable salt according to claim 1 wherein X is selected from 4-10 membered heterocyclyl, optionally further substituted with 1-4 groups selected from C1-6 Alkyl, cyano, C1-6 Cyanoalkyl, C1-6 Substituted by a substituent of hydroxyalkyl; preferably is
    Figure PCTCN2021124884-APPB-100004
    Figure PCTCN2021124884-APPB-100005
    Figure PCTCN2021124884-APPB-100006
    More preferably
    Figure PCTCN2021124884-APPB-100007
  3. The chiral compound, deuterate, or pharmaceutically acceptable salt according to claim 1 or 2, R1 Selected from the group consisting of
    Figure PCTCN2021124884-APPB-100008
    Figure PCTCN2021124884-APPB-100009
    Figure PCTCN2021124884-APPB-100010
    Preferably is
    Figure PCTCN2021124884-APPB-100011
  4. A chiral compound, deuterate, or a pharmaceutically acceptable salt according to any one of claims 1-3 wherein R2 Selected from H, halogen or C1-3 An alkyl group; preferably H.
  5. The chiral compound, deuterate, or pharmaceutically acceptable salt according to any one of claims 1-4 wherein R3 Selected from H, C1-6 Alkyl, substituted C1-6 Alkyl, C2-6 Alkenyl, substituted C2-6 Alkenyl, C3-6 Cycloalkyl or substituted C3-6 Cycloalkyl; preferably C1-3 Alkyl, C3-6 Cycloalkyl or C2-4 Alkenyl groups; more preferably ethyl, vinyl or cyclopropyl.
  6. The chiral compound, deuterate, or pharmaceutically acceptable salt according to any one of claims 1-5 wherein R4 Or R is5 Selected from H, halogen or C1-6 An alkyl group; preferably R4 Is H and R5 Is methyl.
  7. The chiral compound, deuterate, or pharmaceutically acceptable salt according to any one of claims 1-6 wherein R6 Selected from C1-6 Alkoxy, C3-8 Cycloalkyloxy-and cyano-substituted cyclopropyl C1-6 Alkylene oxide group, the C1-6 The alkoxy groups are optionally further substituted with one or more groups selected from halogen, hydroxy, methoxy, C3-8 Substituted cycloalkyl; preferably R6 Selected from the group consisting of
    Figure PCTCN2021124884-APPB-100012
    Figure PCTCN2021124884-APPB-100013
  8. Chiral compound, deuterated or pharmaceutically acceptable salt according to any one of claims 1-7 wherein Y is selected from
    Figure PCTCN2021124884-APPB-100014
    Preferably is
    Figure PCTCN2021124884-APPB-100015
    Figure PCTCN2021124884-APPB-100016
    More preferably
    Figure PCTCN2021124884-APPB-100017
  9. The chiral compound, deuterate, or pharmaceutically acceptable salt according to any one of claims 1-8 wherein R7 Selected from H, cyano, halogen, C1-6 Alkyl, -C0-6 alkylene-ORa 、-C0-6 alkylene-OC (O) N (R)a )2 、-C0-6 alkylene-N (R)a )2 、-C0-6 alkylene-NRa C(O)Ra 、-C0-6 alkylene-NRa C(O)N(Ra )2 、-C0-6 alkylene-NRa S(O)Ra 、-C0-6 alkylene-NRa S(O)2 Ra 、-C0-6 alkylene-S (=o) Ra 、-C0-6 alkylene-S (=o)2 Ra 、-C0-6 alkylene-SRa 、-C0-6 alkylene-S (R)a )5 、-C0-6 alkylene-C (=o) Ra 、-C0-6 alkylene-C (=o) ORa 、-C0-3 alkylene-C (=O) N (R)a )2 Acryl, -C0-6 alkylene-C3-14 Cycloalkyl, -C0-6 Alkylene- (3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 Aryl or-C0-6 Alkylene- (5-14 membered heteroaryl), said C1-6 Alkyl, -C0-6 alkylene-C3-14 Cycloalkyl, -C0-6 Alkylene- (3-14 membered heterocyclyl), -C0-6 alkylene-C6-14 Aryl or-C0-6 Alkylene- (5-14 membered heteroaryl) optionally may also be substituted with 1 or more Ra Substituted; each Ra Each independently selected from H, halogen, hydroxy, amino, oxo, cyano, carboxyl, C1-6 Alkyl, C3-8 Cycloalkyl, 3-8 membered heterocyclyl, C6-14 Aryl, 5-14 membered heteroaryl, C1-6 Acyl or
    Figure PCTCN2021124884-APPB-100018
    Preferably R7 Selected from H, acryl, C1-6 Alkyl, C1-6 Haloalkyl, C1-6 Acyl, -C0-3 alkylene-C1-6 Alkoxy, C1-6 Hydroxyalkyl, C1-6 Cyanoalkyl, -C0-3 alkylene-S (=o)2 -C1-6 Alkyl, C3-6 A ketone group,
    Figure PCTCN2021124884-APPB-100019
    Figure PCTCN2021124884-APPB-100020
    -C0-3 alkylene-C3-6 Cycloalkyl, -C0-3 alkylene-C3-8 Heterocyclyl, -C0-3 Alkylene-phenyl, -C0-3 Alkylene-pyrazolyl, said-C0-3 alkylene-C3-6 Cycloalkyl, -C0-3 alkylene-C3-8 Heterocyclyl, -C0-3 Alkylene-phenyl, -C0-3 The alkylene-pyrazolyl is optionally further substituted with one or more substituents selected from C1-6 Acyl, hydroxy, C1-6 Alkyl, C3-6 Cycloalkyl, halogen,
    Figure PCTCN2021124884-APPB-100021
    Or cyano group, said C3-8 The heterocyclic group contains a heteroatom selected from O or N; more preferably R7 Selected from H, C1-6 Alkyl, acetyl,
    Figure PCTCN2021124884-APPB-100022
    Figure PCTCN2021124884-APPB-100023
  10. The chiral compound, deuterate, or pharmaceutically acceptable salt according to any one of claims 1-9 wherein R8 Selected from H or C1-6 An alkyl group; preferably H.
  11. The chiral compound, deuterated compound, or a pharmaceutically acceptable salt according to any one of claim 1-10, wherein the chiral compound of formula (I) is selected from the group consisting of compounds of formulas (IA) - (ID),
    Figure PCTCN2021124884-APPB-100024
    Figure PCTCN2021124884-APPB-100025
    The method comprises the steps of carrying out a first treatment on the surface of the Preferably is
    Figure PCTCN2021124884-APPB-100026
    Wherein the substituents are as defined in claims 1 to 10.
  12. A chiral compound, deuterate, or a pharmaceutically acceptable salt thereof selected from the group consisting of:
    Figure PCTCN2021124884-APPB-100027
    Figure PCTCN2021124884-APPB-100028
    Figure PCTCN2021124884-APPB-100029
    preferably is
    Figure PCTCN2021124884-APPB-100030
    Figure PCTCN2021124884-APPB-100031
  13. A pharmaceutical composition comprising a chiral compound, deuterated or pharmaceutically acceptable salt according to any one of claims 1-12 and at least one pharmaceutically acceptable excipient, wherein the chiral compound is present in the pharmaceutical composition in an amount greater than the amount of its corresponding axial chiral isomer; preferably, the chiral compound represented by the general formula (I) is contained in the pharmaceutical composition in an amount of more than 51%, 61%, 71%, 81%, 91%, 99% or more in the active ingredient.
  14. Use of a chiral compound, a deuteride, a pharmaceutically acceptable salt, or a pharmaceutical composition according to any one of claims 1-12 in the manufacture of a medicament; preferably, the drug is an anticancer drug; more preferably the cancer is mediated by KRAS G12C, HRAS G12C or NRAS G12C mutations.
  15. A method of treating and/or preventing cancer comprising administering to a subject a therapeutically effective amount of a chiral compound, deuterated, pharmaceutically acceptable salt of any one of claims 1-12 or a pharmaceutical composition of any one of claim 13; preferably the cancer is mediated by KRAS G12C, HRAS G12C or NRAS G12C mutations.
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* Cited by examiner, † Cited by third party
Publication numberPriority datePublication dateAssigneeTitle
US12162893B2 (en)2020-09-232024-12-10Erasca, Inc.Tricyclic pyridones and pyrimidones
AR125782A1 (en)2021-05-052023-08-16Revolution Medicines Inc RAS INHIBITORS
WO2022235870A1 (en)2021-05-052022-11-10Revolution Medicines, Inc.Ras inhibitors for the treatment of cancer
WO2024158778A1 (en)*2023-01-242024-08-02Theras, Inc.Compositions and methods for inhibition of ras

Citations (6)

* Cited by examiner, † Cited by third party
Publication numberPriority datePublication dateAssigneeTitle
CN106488910A (en)*2013-10-102017-03-08亚瑞克西斯制药公司Inhibitors of kras g12c
CN108779097A (en)*2015-11-162018-11-09亚瑞克西斯制药公司Include the quinazoline compound and its application method of the 2- substitutions of substituted heterocycle
CN110267957A (en)*2017-02-022019-09-20安斯泰来制药株式会社 Quinazoline compounds
WO2020113071A1 (en)*2018-11-292020-06-04Araxes Pharma LlcCompounds and methods of use thereof for treatment of cancer
WO2020177629A1 (en)*2019-03-012020-09-10劲方医药科技(上海)有限公司Spiro-substituted pyrimidine-fused cyclic compound, preparation method therefor and medical use thereof
WO2020216190A1 (en)*2019-04-222020-10-29贝达药业股份有限公司Quinazoline compound and pharmaceutical application thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication numberPriority datePublication dateAssigneeTitle
CN112110918B (en)*2019-06-212023-08-22劲方医药科技(上海)有限公司Spiro substituted pyrimido cyclic compounds, process for their preparation and their use in medicine
CN113045570A (en)*2019-12-272021-06-29微境生物医药科技(上海)有限公司Spiro-containing quinazoline compounds

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication numberPriority datePublication dateAssigneeTitle
CN106488910A (en)*2013-10-102017-03-08亚瑞克西斯制药公司Inhibitors of kras g12c
CN108779097A (en)*2015-11-162018-11-09亚瑞克西斯制药公司Include the quinazoline compound and its application method of the 2- substitutions of substituted heterocycle
CN110267957A (en)*2017-02-022019-09-20安斯泰来制药株式会社 Quinazoline compounds
WO2020113071A1 (en)*2018-11-292020-06-04Araxes Pharma LlcCompounds and methods of use thereof for treatment of cancer
WO2020177629A1 (en)*2019-03-012020-09-10劲方医药科技(上海)有限公司Spiro-substituted pyrimidine-fused cyclic compound, preparation method therefor and medical use thereof
WO2020216190A1 (en)*2019-04-222020-10-29贝达药业股份有限公司Quinazoline compound and pharmaceutical application thereof
CN113748114A (en)*2019-04-222021-12-03贝达药业股份有限公司Quinazoline compound and application thereof in medicine

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