WO2022089389A1

WO2022089389A1 - Heterocyclic compound, preparation method therefor, pharmaceutical composition thereof and application thereof

Info

Publication number: WO2022089389A1
Application number: PCT/CN2021/126252
Authority: WO
Inventors: 袁建栋; 方华祥; 黄仰青; 顾家宁; 王晨英; 李宸杰; 吴敬浩
Original assignee: 赣江新区博瑞创新医药有限公司
Priority date: 2020-10-30
Filing date: 2021-10-26
Publication date: 2022-05-05
Also published as: CN114437116A

Abstract

The present invention relates to the field of pharmaceutical chemistry, and relates to a heterocyclic compound, a preparation method therefor, a pharmaceutical composition thereof and an application thereof. In particular, the present invention relates to a heterocyclic compound represented by formula I, a pharmaceutical composition comprising same, and an application thereof as an SHP2 inhibitor in the field of medicine. The heterocyclic compound in the present invention exhibits excellent biological activity and druggability, and has great drug development prospects.

Description

杂环化合物及其制备方法、药物组合物和应用Heterocyclic compound and its preparation method, pharmaceutical composition and application

相关申请的交叉引用CROSS-REFERENCE TO RELATED APPLICATIONS

本发明要求2020年10月30日在中国提交的，名称为“杂环化合物及其制备方法、药物组合物和应用”、申请号为202011196830.8的发明专利申请的优先权，通过引用的方式将该专利申请的全部内容并入本文。The present invention claims the priority of the invention patent application entitled "Heterocyclic compounds and their preparation methods, pharmaceutical compositions and applications" and the application number is 202011196830.8, which was filed in China on October 30, 2020. The entire contents of the patent application are incorporated herein.

技术领域technical field

本发明属于药物化学领域，具体涉及一类杂环化合物、该类化合物及其中间体的制备方法、包含该类化合物的药物组合物及其在医药领域中的应用。The invention belongs to the field of medicinal chemistry, and specifically relates to a class of heterocyclic compounds, a preparation method of the compound and an intermediate thereof, a pharmaceutical composition comprising the compound and its application in the field of medicine.

背景技术Background technique

酪氨酸磷酸酶SHP2由两个N-末端Src同源2结构域(N-SH ₂和C-SH ₂)和一个蛋白酪氨酸磷酸酶催化结构域(PTP)构成。在基础状态下，N-SH ₂能够与PTP结合形成一个环状结构，从而阻碍PTP与底物的结合，使得酶催化活性被抑制；当上游受体蛋白的酪氨酸被磷酸化后，N-SH ₂与之相结合，PTP催化域得到释放，从而发挥出磷酸酶活性。 The tyrosine phosphatase SHP2 consists of two N-terminal Src homology 2 domains (N-SH ₂ and C-SH ₂ ) and a protein tyrosine phosphatase catalytic domain (PTP). In the basal state, N-SH ₂ can combine with PTP to form a ring structure, thereby hindering the binding of PTP and the substrate, so that the catalytic activity of the enzyme is inhibited; when the tyrosine of the upstream receptor protein is phosphorylated, N -SH ₂ binds to it, the PTP catalytic domain is released, and the phosphatase activity is exerted.

在细胞水平上，SHP2通过在诸多受体酪氨酸激酶的细胞质下游的功能作用，参与多个肿瘤细胞信号传导通路，如RTK/Ras/MAPK、JAK/STAT和PB3K/Akt等。通过对这些激酶以及信号通路的调控作用，SHP2与许多重要的细胞生命活动密切相关，如细胞增殖、迁移、分化、死亡、细胞因子的调控及肿瘤发生等等。At the cellular level, SHP2 participates in multiple tumor cell signaling pathways, such as RTK/Ras/MAPK, JAK/STAT, and PB3K/Akt, by functioning downstream of many receptor tyrosine kinases in the cytoplasm. Through the regulation of these kinases and signaling pathways, SHP2 is closely related to many important cell life activities, such as cell proliferation, migration, differentiation, death, regulation of cytokines, and tumorigenesis.

同时，SHP2也参与程序性死亡受体1(PD1)介导的免疫***抑制。T细胞的PD-1与PD-L1结合后，在细胞内能够招募大量的SHP2。SHP2能够将T细胞内抗原受体通路蛋白去磷酸化，从而抑制T细胞的激活。因此，抑制SHP2的活性能够在肿瘤微环境中逆转免疫抑制。Meanwhile, SHP2 is also involved in programmed death receptor 1 (PD1)-mediated suppression of the immune system. After the PD-1 of T cells binds to PD-L1, a large amount of SHP2 can be recruited in the cell. SHP2 can dephosphorylate antigen receptor pathway proteins in T cells, thereby inhibiting T cell activation. Therefore, inhibition of SHP2 activity could reverse immunosuppression in the tumor microenvironment.

作为一类重要的细胞信号因子，SHP2突变与多种疾病密切相关。研究发现，在神经母细胞瘤、AML(4％)、乳腺癌、NSCLC(10％)、肺腺癌(30％)、食道癌、头颈部肿瘤、黑色素瘤和胃癌中存在SHP2突变。As an important cell signaling factor, SHP2 mutation is closely related to a variety of diseases. The study found SHP2 mutations in neuroblastoma, AML (4%), breast cancer, NSCLC (10%), lung adenocarcinoma (30%), esophageal cancer, head and neck tumors, melanoma and gastric cancer.

目前已有多个SHP2变构抑制剂进入临床研究阶段，如Novartis公司开发的TNO-155，Revolution Medicine公司开发的RMC-4630，以及北京加科思的JAB-3068等化合物都已经进入临床研究阶段，但还没有一款开发上市的SHP2抑制剂，用于制备治疗努南综合征、豹皮综合征、白血病、成神经细胞瘤、黑色素瘤、乳腺癌、食道癌、头颈部肿瘤、肺癌及结肠癌。因此，迫切需要开发一类成药性好的SHP2抑制剂药物。At present, a number of SHP2 allosteric inhibitors have entered the clinical research stage, such as TNO-155 developed by Novartis, RMC-4630 developed by Revolution Medicine, and JAB-3068 developed by Beijing Jiakesi and other compounds have entered the clinical research stage , but there is no SHP2 inhibitor that has been developed and marketed for the preparation and treatment of Noonan syndrome, leopard skin syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, head and neck cancer, lung cancer and Colon cancer. Therefore, there is an urgent need to develop a class of SHP2 inhibitors with good druggability.

发明内容SUMMARY OF THE INVENTION

发明要解决的问题Invention to solve problem

本发明旨在提供一类全新的用作SHP2抑制剂的杂环化合物，其表现出对肿瘤细胞很好的抑制活性，且成药性好，具有广阔的药物开发前景。而且，该类化合物的制备方法简单，有利于工业化生产。The present invention aims to provide a new class of heterocyclic compounds used as SHP2 inhibitors, which exhibits good inhibitory activity on tumor cells, has good druggability, and has broad prospects for drug development. Moreover, the preparation method of the compound is simple, which is favorable for industrial production.

用于解决问题的方案solution to the problem

第一方面，本发明提供了一种如式I所示的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药，其中In a first aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, wherein

X ¹、X ²和X ³各自独立地选自CR ₅和N，或不存在； X ¹ , X ² and X ³ are each independently selected from CR ₅ and N, or are absent;

当X ¹、X ²和X ³各自独立地选自CR ₅和N时，X ⁵和X ⁶各自独立地选自C和N，X ⁷为CR ₅或N； When X ¹ , X ² and X ³ are each independently selected from CR ₅ and N, X ⁵ and X ⁶ are each independently selected from C and N, and X ⁷ is CR ₅ or N;

当X ¹、X ²和X ³不存在时，X ⁵、X ⁶和X ⁷各自独立地选自CR ₅和N； When X ¹ , X ² and X ³ are absent, X ⁵ , X ⁶ and X ⁷ are each independently selected from CR ₅ and N;

X ⁴为C或N； X ⁴ is C or N;

X ⁸为N或NR ₅； X ⁸ is N or NR ₅ ;

R ₁、R ₂、R ₃、R ₄、R ₆和R ₇各自独立地选自氢、卤素、羟基、氨基、氧代基、氰基、C ₂-C ₈烯基、C ₂-C ₈炔基、醛基、氨基甲酰基、C ₁-C ₈烷基、C ₁-C ₈杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₈烷氧基和C ₁-C ₃卤代烷氧基，其中所述C ₁-C ₈烷基、C ₁-C ₈杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₈烷氧基和C ₁-C ₃卤代烷氧基各自任选地被一个或多个R ₅取代； R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R ₇ are each independently selected from hydrogen, halogen, hydroxy, amino, oxo, cyano, C ₂ -C ₈ alkenyl, C ₂ -C ₈ Alkynyl, aldehyde, carbamoyl, C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₈ alkoxy and C ₁ -C ₃ haloalkoxy, wherein said C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycle alkyl, _C1 - _C8alkoxy and _C1 _- C3 haloalkoxy are each optionally substituted with one or more R5 _;

若存在，每一个R ₅各自独立地选自氢、卤素、羟基、氨基、氰基、氨基甲酰基、C ₁-C ₃烷基、C ₁-C ₃杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₃烷氧基和C ₁-C ₃卤代烷氧基，其中所述C ₁-C ₃烷基、C ₁-C ₃杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₃烷氧基和C ₁-C ₃卤代烷氧基各自任选地被一个或多个R ₈取代； If present, each R ₅ is independently selected from hydrogen, halogen, hydroxy, amino, cyano, carbamoyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ heteroalkyl, C ₃ -C ₈ ring Alkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₃ alkoxy and C ₁ -C ₃ haloalkoxy, wherein said C ₁ -C ₃ alkyl, C ₁ -C ₃ heteroalkyl , C3 _- _C8cycloalkyl , C3- _{C8heterocycloalkyl} , _C1 _- _C3alkoxy , and _C1 _- C3haloalkoxy are each optionally substituted with one or more _R8 ;

若存在，每一个R ₈各自独立地选自氢、卤素、羟基、氨基和氰基； If present, each R is _{independently} selected from hydrogen, halogen, hydroxy, amino and cyano;

A和B各自独立地选自C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₆-C ₁₀芳基和C ₅-C ₁₂杂芳基； A and B are each independently selected from C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl, and C ₅ -C ₁₂ heteroaryl;

其中所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3； wherein the heteroatoms or heteroatomic groups in the heteroalkyl, heterocycloalkyl and heteroaryl groups are each independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S -, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O) NH-, the number of heteroatoms or heteroatoms in the heteroalkyl, heterocycloalkyl and heteroaryl groups is each independently selected from 1, 2 and 3;

n为1、2或3。n is 1, 2 or 3.

具体地，如式I所示的化合物为如式I-1或式I-2所示的化合物，其中Specifically, the compound represented by formula I is the compound represented by formula I-1 or formula I-2, wherein

X ⁵、X ⁶和X ⁷各自独立地选自CR ₅和N； X ⁵ , X ⁶ and X ⁷ are each independently selected from CR ₅ and N;

X ⁸、R ₁、R ₂、R ₃、R ₄、R ₅、R ₆、B和n如式I所定义。 X ⁸ , R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , B and n are as defined in formula I.

优选地，在如式I-1或式I-2所示的化合物中，Preferably, in the compound shown in formula I-1 or formula I-2,

X ⁸为N或NR ₅； X ⁸ is N or NR ₅ ;

R ₁为氢、氨基、氧代基或C ₁-C ₈烷基，其中所述C ₁-C ₈烷基任选地被一个或多个R ₅取代； R ₁ is hydrogen, amino, oxo or C ₁ -C ₈ alkyl, wherein said C ₁ -C ₈ alkyl is optionally substituted with one or more R ₅ ;

R ₂为氢； R ₂ is hydrogen;

R ₃为氢或羟基； R ₃ is hydrogen or hydroxyl;

每一个R ₄各自独立地选自氢、卤素和C ₁-C ₈烷基，其中所述C ₁-C ₈烷基任选地被一个或多个R ₅取代； Each R ₄ is independently selected from hydrogen, halogen and C ₁ -C ₈ alkyl, wherein said C ₁ -C ₈ alkyl is optionally substituted with one or more R ₅ ;

R ₆为氢； R ₆ is hydrogen;

若存在，每一个R ₅各自独立地选自氢、氨基、氨基甲酰基、C ₁-C ₃烷基和C ₁-C ₃杂烷基，其中所述C ₁-C ₃烷基、C ₁-C ₃杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₃烷氧基和C ₁-C ₃卤代烷氧基各自任选地被一个或多个R ₈取代；优选地，若存在，每一个R ₅各自独立地选自氢、氨基、氨基甲酰基和C ₁-C ₃烷基，其中所述C ₁-C ₃烷基任选地被一个或多个R ₈取代； If present, each R ₅ is independently selected from hydrogen, amino, carbamoyl, C ₁ -C ₃ alkyl and C ₁ -C ₃ heteroalkyl, wherein said C ₁ -C ₃ alkyl, C ₁ _-C3 -heteroalkyl, C3 _- _C8cycloalkyl , C3- _{C8heterocycloalkyl} , _C1 _- _C3alkoxy , and _C1 _- C3haloalkoxy are each optionally replaced by one or Multiple R ₈ substitutions; preferably, if present, each R ₅ is independently selected from hydrogen, amino, carbamoyl, and C ₁ -C ₃ alkyl, wherein said C ₁ -C ₃ alkyl is optionally is substituted by one or more R ₈ ;

若存在，每一个R ₈各自独立地选自氢和羟基； If present, each R is _{independently} selected from hydrogen and hydroxyl;

B为C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₆-C ₁₀芳基和C ₅-C ₁₂杂芳基； B is C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl and C ₅ -C ₁₂ heteroaryl;

其中所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3；优选地，所述杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3； wherein the heteroatoms or heteroatomic groups in the heteroalkyl, heterocycloalkyl and heteroaryl groups are each independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S -, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O) NH-, the number of heteroatoms or heteroatom groups in the heteroalkyl, heterocycloalkyl and heteroaryl groups is each independently selected from 1, 2 and 3; preferably, the heterocycloalkyl and heteroaryl groups The heteroatoms or heteroatomic groups in are independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S-, -C(=O)O-, -C(= O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O)NH-, among the heterocycloalkyl and heteroaryl the number of heteroatoms or heteroatomic groups is each independently selected from 1, 2 and 3;

n为1、2或3。n is 1, 2 or 3.

更优选地，在如式I-1或式I-2所示的化合物中，More preferably, in the compound shown in formula I-1 or formula I-2,

结构中的

片段选自下列片段中的任意一种： in structure

Fragments are selected from any of the following fragments:

甚至更优选地，结构中的

片段选自下列片段中的任意一种： Even more preferably, in the structure

Fragments are selected from any of the following fragments:

或者甚至更优选地，结构中的

片段选自下列片段中的任意一种： Or even more preferably, in the structure

Fragments are selected from any of the following fragments:

具体地，如式I所示的化合物为如式I-3或式I-4所示的化合物，其中Specifically, the compound shown in formula I is the compound shown in formula I-3 or formula I-4, wherein

X ¹、X ²和X ³各自独立地选自CR ₅或N，X ⁵和X ⁶各自独立地选自C和N，X ⁷为CR ₅或N； X ¹ , X ² and X ³ are each independently selected from CR ₅ or N, X ⁵ and X ⁶ are each independently selected from C and N, and X ⁷ is CR ₅ or N;

优选地，在如式I-3或式I-4所示的化合物中，Preferably, in the compound shown in formula I-3 or formula I-4,

X ⁸为N或NR ₅； X ⁸ is N or NR ₅ ;

R ₁为氢或C ₁-C ₈烷基，其中所述C ₁-C ₈烷基任选地被一个或多个R ₅取代； R ₁ is hydrogen or C ₁ -C ₈ alkyl, wherein said C ₁ -C ₈ alkyl is optionally substituted with one or more R ₅ ;

R ₂为氢； R ₂ is hydrogen;

R ₃氢或羟基； R ₃ hydrogen or hydroxyl;

R ₆为氢； R ₆ is hydrogen;

若存在，每一个R ₅各自独立地选自氢和C ₁-C ₃烷基； If present, each R ₅ is independently selected from hydrogen and C ₁ -C ₃ alkyl;

其中所述杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3； wherein the heteroatoms or heteroatomic groups in the heterocycloalkyl and heteroaryl groups are each independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S-, -C (=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O)NH-, so The number of heteroatoms or heteroatomic groups in the heterocycloalkyl and heteroaryl groups is each independently selected from 1, 2 and 3;

n为1、2或3。n is 1, 2 or 3.

更优选地，在如式I-3或式I-4所示的化合物中，More preferably, in the compound shown in formula I-3 or formula I-4,

结构中的

片段选自下列片段中的任意一种： in structure

Fragments are selected from any of the following fragments:

甚至更优选地，结构中的

Fragments are selected from any of the following fragments:

进一步优选地，在如式I-1、式I-2、式I-3或式I-4所示的化合物中，Further preferably, in the compound shown in formula I-1, formula I-2, formula I-3 or formula I-4,

结构中的

片段选自下列片段中的任意一种： in structure

Fragments are selected from any of the following fragments:

甚至进一步优选地，结构中的

片段选自下列片段中的任意一种： Even further preferably, in the structure

Fragments are selected from any of the following fragments:

第二方面，本发明提供了如式I、式I-1、式I-2、式I-3或式I-4所示的具体化合物，其选自：In the second aspect, the present invention provides a specific compound as shown in formula I, formula I-1, formula I-2, formula I-3 or formula I-4, which is selected from:

(1)(S)-N-(3-(3-氨基-5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)吡嗪-2-基硫基)-2-氯苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(1) (S)-N-(3-(3-Amino-5-(5-amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]) -1'-yl)pyrazin-2-ylsulfanyl)-2-chlorophenyl)-2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1, 2-a]pyrimidine-3-carboxamide;

(2)(S)-N-(3-(5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)咪唑并[1,2-c]嘧啶-8-基硫基)-2-氯苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(2) (S)-N-(3-(5-(5-Amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]-1'- yl)imidazo[1,2-c]pyrimidin-8-ylsulfanyl)-2-chlorophenyl)-2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H-pyridine and [1,2-a]pyrimidine-3-carboxamide;

(3)(S)-N-(3-(5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)-7-甲基咪唑并[1,2-c]嘧啶-8-基硫基)-2-氯苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(3) (S)-N-(3-(5-(5-Amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]-1'- yl)-7-methylimidazo[1,2-c]pyrimidin-8-ylsulfanyl)-2-chlorophenyl)-2-hydroxy-4-oxo-6,7,8,9-tetra Hydrogen-4H-pyrido[1,2-a]pyrimidine-3-carboxamide;

(4)(S)-N-(3-(4-氨基-2-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)-1-甲基-6-氧代-1,6-二氢嘧啶-5-基硫基)-2-氯苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(4) (S)-N-(3-(4-Amino-2-(5-amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]) -1'-yl)-1-methyl-6-oxo-1,6-dihydropyrimidin-5-ylsulfanyl)-2-chlorophenyl)-2-hydroxy-4-oxo-6, 7,8,9-Tetrahydro-4H-pyrido[1,2-a]pyrimidine-3-carboxamide;

(5)(S)-N-(3-(3-氨基-5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)吡嗪-2-基硫基)-2-氯苯基)-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(5) (S)-N-(3-(3-Amino-5-(5-amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]) -1'-yl)pyrazin-2-ylsulfanyl)-2-chlorophenyl)-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-a] Pyrimidine-3-carboxamide;

(6)(S)-N-(3-(3-氨基-5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)吡嗪-2-基硫基)-2-甲基苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(6) (S)-N-(3-(3-Amino-5-(5-amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]) -1'-yl)pyrazin-2-ylsulfanyl)-2-methylphenyl)-2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1 ,2-a]pyrimidine-3-carboxamide;

(7)(S)-N-(3-(3-氨基-5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)吡嗪-2-基硫基)-5-氟-2-甲基苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(7) (S)-N-(3-(3-Amino-5-(5-amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]) -1'-yl)pyrazin-2-ylthio)-5-fluoro-2-methylphenyl)-2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H- pyrido[1,2-a]pyrimidine-3-carboxamide;

(8)(S)-3-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)-6-(2-氯-3-(2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺基)苯基硫基)吡嗪-2-甲酸乙酯；(8) (S)-3-(5-Amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidin]-1'-yl)-6-( 2-Chloro-3-(2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]pyrimidine-3-carboxamido)phenylthio ) ethyl pyrazine-2-carboxylate;

(9)(S)-3-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)-6-(2-氯-3-(2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺基)苯基硫基)吡嗪-2-甲酸；(9) (S)-3-(5-Amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidin]-1'-yl)-6-( 2-Chloro-3-(2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]pyrimidine-3-carboxamido)phenylthio ) pyrazine-2-carboxylic acid;

(10)(S)-N-(3-(5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)-6-氨基甲酰基吡嗪-2-基硫基)-2-氯苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺；(10) (S)-N-(3-(5-(5-Amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]-1'- yl)-6-carbamoylpyrazin-2-ylthio)-2-chlorophenyl)-2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[ 1,2-a]pyrimidine-3-carboxamide;

(11)(S)-3-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)-6-(2-氯-3-(2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺基)苯基硫基)-5-甲基吡嗪-2-甲酸乙酯；和(11) (S)-3-(5-Amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidin]-1'-yl)-6-( 2-Chloro-3-(2-hydroxy-4-oxo-6,7,8,9-tetrahydro-4H-pyrido[1,2-a]pyrimidine-3-carboxamido)phenylthio )-ethyl 5-methylpyrazine-2-carboxylate; and

(12)(S)-N-(3-(5-(5-氨基-5,7-二氢螺[环戊二烯并[b]吡啶-6,4'-哌啶]-1'-基)-6-氨基甲酰基-3-甲基吡嗪-2-基硫基)-2-氯苯基)-2-羟基-4-氧代-6,7,8,9-四氢-4H-吡啶并[1,2-a]嘧啶-3-甲酰胺。(12) (S)-N-(3-(5-(5-Amino-5,7-dihydrospiro[cyclopentadieno[b]pyridine-6,4'-piperidine]-1'- yl)-6-carbamoyl-3-methylpyrazin-2-ylthio)-2-chlorophenyl)-2-hydroxy-4-oxo-6,7,8,9-tetrahydro- 4H-pyrido[1,2-a]pyrimidine-3-carboxamide.

第三方面，本发明提供了如式I所述的化合物的制备方法，其包括下列步骤：In a third aspect, the present invention provides the preparation method of the compound as described in formula I, which comprises the following steps:

1)化合物A与化合物B反应，得到化合物C；1) compound A reacts with compound B to obtain compound C;

2)化合物C与化合物D反应，得到化合物E；和2) compound C reacts with compound D to obtain compound E; and

3)化合物E与化合物F反应，得到目标产物；3) compound E reacts with compound F to obtain the target product;

其中in

LG ₁和LG ₂各自独立地选自氯和溴，LG ₃为氯、溴或羟基； LG ₁ and LG ₂ are each independently selected from chlorine and bromine, and LG ₃ is chlorine, bromine or hydroxyl;

X ¹、X ²、X ³、X ⁴、X ⁵、X ⁶、X ⁷、X ⁸、R ₁、R ₂、R ₃、R ₄、R ₆、R ₇、A、B和n如式I所定义。 X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ , A, B and n are as in formula I defined.

在上述制备方法中，化合物A和化合物B可以在催化剂(例如钯催化剂，如Pd ₂(dba) ₃)、配体(例如Xantphos)和碱(例如有机碱，如DIEA；又如无机碱，如碳酸铯)存在的条件下反应，得到化合物C；化合物C和化合物D可以在碱和缩合剂(例如HATU、HBTU、EDCT/HOBT等)存在的条件下反应(例如缩合反应)，得到化合物E；化合物E和化合物F在碱(例如有机碱，如DIEA；又如无机碱，如碳酸钾、碳酸铯、叔丁醇钠等)存在的条件下反应，得到式I化合物。 In the above preparation methods, compound A and compound B can be prepared in the presence of catalysts (eg, palladium catalysts, such as Pd ₂ (dba) ₃ ), ligands (eg, Xantphos) and bases (eg, organic bases, such as DIEA; and inorganic bases, such as cesium carbonate) to obtain compound C; compound C and compound D can react (such as condensation reaction) in the presence of a base and a condensing agent (such as HATU, HBTU, EDCT/HOBT, etc.) to obtain compound E; Compound E and compound F are reacted in the presence of a base (eg, an organic base, such as DIEA; another example, an inorganic base, such as potassium carbonate, cesium carbonate, sodium tert-butoxide, etc.) to obtain the compound of formula I.

本发明对于上述制备方法中各步骤所使用的反应溶剂没有特别限制，任何在一定程度上能够溶解起始原料并且不抑制反应的溶剂(例如DMF、DMSO、NMP等)均包含在本发明的范围之中。另外，本领域的许多类似改动或等同替换或者相应的溶剂组合，均视为包含在本发明的范围之中。The present invention has no particular limitation on the reaction solvent used in each step of the above preparation method, and any solvent (such as DMF, DMSO, NMP, etc.) that can dissolve the starting materials to a certain extent and does not inhibit the reaction is included in the scope of the present invention among. In addition, many similar modifications or equivalent substitutions in the art or corresponding solvent combinations are deemed to be included within the scope of the present invention.

第四方面，本发明提供了一种药物组合物，其包含如式I、式I-1、式I-2、式I-3或式I-4所示的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药，和至少一种药学上可接受的辅料。In a fourth aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I, formula I-1, formula I-2, formula I-3 or formula I-4 or a pharmaceutically acceptable compound thereof A salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug, and at least one pharmaceutically acceptable excipient.

第五方面，本发明提供了如式I、式I-1、式I-2、式I-3或式I-4所示的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药或者包含其的药物组合物，其用作SHP2抑制剂或者用于预防和/或治疗与SHP2活性异常相关的疾病或病症。In the fifth aspect, the present invention provides a compound represented by formula I, formula I-1, formula I-2, formula I-3 or formula I-4 or a pharmaceutically acceptable salt, hydrate, solvate, Stereoisomers, tautomers, metabolites or prodrugs or pharmaceutical compositions comprising the same for use as SHP2 inhibitors or for the prevention and/or treatment of diseases or conditions associated with abnormal SHP2 activity.

第六方面，本发明提供了如式I、式I-1、式I-2、式I-3或式I-4所示的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药或者包含其的药物组合物在制备用于预防和/或治疗与SHP2活性异常相关的疾病或病症的药物中的应用。In the sixth aspect, the present invention provides a compound represented by formula I, formula I-1, formula I-2, formula I-3 or formula I-4 or a pharmaceutically acceptable salt, hydrate, solvate, Use of a stereoisomer, tautomer, metabolite or prodrug or a pharmaceutical composition comprising the same in the preparation of a medicament for preventing and/or treating a disease or condition associated with abnormal SHP2 activity.

第七方面，本发明提供了一种用于预防和/或治疗与SHP2活性异常相关的疾病或病症的方法，其包括将预防和/或治疗有效量的如式I、式I-1、式I-2、式I-3或式I-4所示的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药或者包含其的药物组合物施用于对其有需要的个体。In the seventh aspect, the present invention provides a method for preventing and/or treating a disease or condition related to abnormal SHP2 activity, which comprises preventing and/or treating an effective amount of formula I, formula I-1, formula I-2, a compound represented by formula I-3 or formula I-4 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof Pharmaceutical compositions thereof are administered to an individual in need thereof.

优选地，在上述医药用途中，所述与SHP2活性异常相关的疾病或病症选自努南综合征、豹皮综合征、白血病、成神经细胞瘤、黑色素瘤、乳腺癌、食道癌、肺癌、结肠癌、头颈部肿瘤、胃癌、间变性大细胞淋巴瘤和成胶质细胞瘤，优选非小细胞肺癌、食管癌和头颈部肿瘤。Preferably, in the above medical use, the disease or condition associated with abnormal SHP2 activity is selected from Noonan syndrome, leopard skin syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, lung cancer, Colon cancer, head and neck tumors, gastric cancer, anaplastic large cell lymphoma and glioblastoma, preferably non-small cell lung cancer, esophageal cancer and head and neck tumors.

发明的效果effect of invention

本发明中的杂环化合物是一类新颖的变构抑制剂，能够通过与SHP2非催化区域的结合并“锁”住SHP2活性很弱的基础状态，从而达到抑制其活性的目的。本发明中的杂环化合物克服了PTP催化区域抑制剂普遍存在的选择性和成药性差等缺点，表现出优良的生物活性及可成药性，极具药物开发前景。The heterocyclic compound in the present invention is a novel allosteric inhibitor, which can inhibit the activity of SHP2 by combining with the non-catalytic region of SHP2 and "locking" the weak basic state of SHP2 activity. The heterocyclic compound in the present invention overcomes the common shortcomings of PTP catalytic region inhibitors such as selectivity and poor druggability, exhibits excellent biological activity and druggability, and has great prospects for drug development.

另外，在相同条件的SHP2酶活性抑制实验、磷酸化蛋白激酶(p-ERK)细胞实验、NCI-H358细胞抗增殖实验、MV-4-11细胞抗增殖实验等评价体系中，与已知化合物RMC4550及TNO155相比，本发明的化合物表现出更优越的药理活性及药代性质。In addition, in the same conditions of the SHP2 enzyme activity inhibition test, phosphorylated protein kinase (p-ERK) cell test, NCI-H358 cell anti-proliferation test, MV-4-11 cell anti-proliferation test and other evaluation systems, and known compounds. Compared with RMC4550 and TNO155, the compounds of the present invention show more superior pharmacological activity and pharmacokinetic properties.

具体实施方式Detailed ways

一般术语和定义General terms and definitions

除非有相反陈述，否则在本发明中所使用的术语具有下述含义。Unless stated to the contrary, terms used in the present invention have the following meanings.

“烷基”是指饱和的脂族烃基团，包括1至20个碳原子的直链和支链基团，例如可以是1至18个碳原子、1至12个碳原子、1至8个碳原子、1至6个碳原子或1至4个碳原子的直链和支链基团。在本发明中，“烷基”可以是一价的、二价的或三价基团。非限制性实例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基及其各种支链异构体等。非限制性实例还包括但不限于亚甲基、次甲基、亚乙基、次乙基、亚丙基、次丙基、亚丁基、次丁基及其各种支链异构体。另外，在本发明中，“烷基”可以是任选取代的或未取代的。"Alkyl" refers to saturated aliphatic hydrocarbon groups, including straight and branched chain groups of 1 to 20 carbon atoms, for example, may be 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms carbon atoms, straight and branched chain groups of 1 to 6 carbon atoms or 1 to 4 carbon atoms. In the present invention, "alkyl" may be a monovalent, divalent or trivalent group. Non-limiting examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl -2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl and various branched chain isomers, etc. Non-limiting examples also include, but are not limited to, methylene, methine, ethylene, ethylene, propylene, propylene, butylene, butylene, and various branched chain isomers thereof. In addition, in the present invention, "alkyl" may be optionally substituted or unsubstituted.

“杂烷基”是指饱和的脂族烃基团，包括1至20个碳原子的直链和支链基团，例如可以是1至18个碳原子、1至12个碳原子、1至8个碳原子、1至6个碳原子或1至4个碳原子的直链和支链基团，上述直链或支链碳链被一个或多个(例如，1个、2个、3个等)杂原子或杂原子团间隔，并通过杂原子或杂原子团连接，上述杂原子或杂原子团选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-。在本发明中，“杂烷基”可以是一价的、二价的或三价基团。非限制性实例包括但不限于被-C(＝O)O-间隔的C ₁-C ₃杂烷基，例如羧基(C ₁杂烷基)、甲氧羰基(C ₂杂烷基)、乙氧羰基(C ₃杂烷基)等，或者被-C(＝O)NH-间隔的C ₁-C ₃杂烷基，例如氨基甲酰基(C ₁杂烷基)、甲氨基甲酰基(C ₂杂烷基)、乙氨基甲酰基(C ₃杂烷基)等。另外，在本发明中，“杂烷基”可以是任选取代的或未取代的。 "Heteroalkyl" means a saturated aliphatic hydrocarbon group, including straight and branched chain groups of 1 to 20 carbon atoms, for example, may be 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms linear and branched groups of 1 to 6 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms, wherein the linear or branched carbon chain is separated by one or more (eg, 1, 2, 3 etc.) interspersed by heteroatoms or heteroatomic groups and connected by heteroatoms or heteroatomic groups selected from -C(=O)NH-, -NH-, -N=, -O-, -S- , -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O)NH -. In the present invention, "heteroalkyl" may be a monovalent, divalent or trivalent group. Non-limiting examples include, but are not limited to, _C1 _- C3 heteroalkyl interrupted by -C(=O)O-, such as carboxy ( _C1 heteroalkyl), methoxycarbonyl ( _C2 heteroalkyl), ethyl Oxycarbonyl (C ₃ heteroalkyl), etc., or C ₁ -C ₃ heteroalkyl interrupted by -C(=O)NH-, such as carbamoyl (C ₁ heteroalkyl), methylcarbamoyl (C ₂ heteroalkyl), ethylcarbamoyl (C ₃ heteroalkyl) and the like. In addition, in the present invention, "heteroalkyl" may be optionally substituted or unsubstituted.

“环烷基”是指饱和或部分不饱和的、单环或多环的脂族烃基团，包括3至12个环原子，例如可以是3至12个、3至10个或3至6个环原子(即3至6元环)。单环环烷基的非限制性实例包括但不限于环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基、环庚三烯基、环辛基等。在本发明中，“环烷基”可以是任选取代的或未取代的。"Cycloalkyl" means a saturated or partially unsaturated, monocyclic or polycyclic, aliphatic hydrocarbon group comprising 3 to 12 ring atoms, eg, 3 to 12, 3 to 10, or 3 to 6 Ring atoms (ie, 3 to 6 membered rings). Non-limiting examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclopentyl Heptatrienyl, cyclooctyl, etc. In the present invention, "cycloalkyl" may be optionally substituted or unsubstituted.

“杂环烷基”是指饱和或部分不饱和的、单环或多环的脂族烃基团，包括3至20个环原子，例如可以是3至16个、3至12个、3至10个或3至6个环原子，其中一个或多个环原子为选自氮、氧或S(O) _m(其中m为0、1或2)的杂原子，其余环原子为碳。优选杂环烷基包括3至12个环原子，其中1至4个环原子是杂原子，更优选包括3至10个环原子，最优选包括5或6个环原子，其中1至4个，优选1至3个，更优选1至2个是杂原子。单环杂环烷基的非限制性实例包括但不限于吡咯烷基、哌啶基、哌嗪基、吗啉基、硫代吗啉基、高哌嗪基等。多环杂环烷基的非限制性实例包括但不限于螺环或桥环的杂环烷基。 "Heterocycloalkyl" refers to a saturated or partially unsaturated, monocyclic or polycyclic aliphatic hydrocarbon group comprising 3 to 20 ring atoms, for example, may be 3 to 16, 3 to 12, 3 to 10 or 3 to 6 ring atoms, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (where m is 0, 1 or 2) and the remaining ring atoms are carbon. Preferred heterocycloalkyl groups comprise 3 to 12 ring atoms, of which 1 to 4 are heteroatoms, more preferably 3 to 10 ring atoms, and most preferably 5 or 6 ring atoms, of which 1 to 4, Preferably 1 to 3, more preferably 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocycloalkyl include, but are not limited to, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and the like. Non-limiting examples of polycyclic heterocycloalkyl groups include, but are not limited to, spirocyclic or bridged ring heterocycloalkyl groups.

“卤素”是指氟、氯、溴和碘，优选氟、氯和溴。"Halogen" means fluorine, chlorine, bromine and iodine, preferably fluorine, chlorine and bromine.

“任选”或“任选地”意味着随后所描述地事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生的情形。例如，“任选被烷基取代的杂环基团”意味着烷基可以但并非必须存在，该说明包括杂环基团被烷基取代的情形和杂环基团不被烷基取代的情形。"Optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance does or does not occur. For example, "heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may, but need not be, present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group .

“取代的”是指基团中的一个或多个氢原子，优选最多5个，更优选1至3个氢原子彼此独立地被相应数目的取代基取代。"Substituted" means that one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3 hydrogen atoms, independently of one another, are substituted by the corresponding number of substituents.

“药学上可接受的盐”是指由本发明中的化合物与相对无毒的酸或碱制备得到的盐。"Pharmaceutically acceptable salts" refers to salts prepared from compounds of the present invention with relatively non-toxic acids or bases.

“药物组合物”是指可供药用的组合物，其包含一种或多种如式I所示的化合物或其药学上可接受的形式(例如盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物、前药等)，以及其他组分(例如药学上可接受的辅料)。"Pharmaceutical composition" refers to a pharmaceutically acceptable composition comprising one or more compounds of Formula I or a pharmaceutically acceptable form thereof (eg, a salt, hydrate, solvate, stereoisomer isomers, tautomers, metabolites, prodrugs, etc.), and other components (eg, pharmaceutically acceptable excipients).

在本发明中，“药学上可接受的辅料”是指在药物生产领域中广泛采用的辅助物料。使用辅料的主要目的在于提供一种使用安全、性质稳定和/或具有特定功能性的药物组合物，还在于提供一种方法，以便在为受试者施用药物之后，活性成分能够以所期望的速率溶出，或者促进活性成分在接受给药的受试者体内得到有效吸收。药学上可接受的辅料可以是具有惰性的填充剂，也可以是为药用组合物提供某种功能(例如稳定组合物的整体pH值或防止组合物中活性成分的降解)的功效成分。药学上可接受的辅料的非限制性实例包括但不限于粘合剂、助悬剂、乳化剂、稀释剂(或填充剂)、成粒剂、胶粘剂、崩解剂、润滑剂、抗粘着剂、助流剂、润湿剂、胶凝剂、吸收延迟剂、溶解抑制剂、增强剂、吸附剂、缓冲剂、螯合剂、防腐剂、着色剂、矫味剂、甜味剂等。In the present invention, "pharmaceutically acceptable auxiliary materials" refer to auxiliary materials widely used in the field of pharmaceutical production. The main purpose of using excipients is to provide a pharmaceutical composition that is safe to use, stable in properties and/or has specific functionality, and also to provide a method so that after the drug is administered to a subject, the active ingredient can be The rate of dissolution, or the promotion of effective absorption of the active ingredient in the subject to which it is administered. Pharmaceutically acceptable excipients can be inert fillers or functional ingredients that provide a certain function for the pharmaceutical composition (eg, stabilizing the overall pH of the composition or preventing the degradation of active ingredients in the composition). Non-limiting examples of pharmaceutically acceptable adjuvants include, but are not limited to, binders, suspending agents, emulsifiers, diluents (or fillers), granulating agents, sizing agents, disintegrating agents, lubricants, anti-adhering agents , glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavors, sweeteners, etc.

本发明中的药物组合物可以使用本领域技术人员已知的任何方法来制备。例如，常规混合、溶解、造粒、乳化、磨细、包封、包埋和/或冻干工艺。The pharmaceutical compositions of the present invention can be prepared using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, attenuating, encapsulating, entrapping and/or lyophilizing processes.

在本发明中，使用药物组合物的目的在于促进针对生物体的给药，有利于活性成分的吸收，进而发挥生物活性。本发明的药物组合物可以通过任何形式给药，包括注射(动脉内、静脉内、肌肉内、腹膜内、皮下)、粘膜、口服(口服固体制剂、口服液体制剂)、直肠、吸入、植入、局部(例如眼部)给药等。口服固体制剂的非限制性实例包括但不限于散剂、胶囊剂、锭剂、颗粒剂、片剂等。口服或粘膜给药的液体制剂的非限制性实例包括但不限于混悬剂、酊剂、酏剂、溶液剂等。局部给药制剂的非限制性实例包括但不限于乳剂、凝胶剂、软膏剂、乳膏剂、贴剂、糊剂、泡沫剂、洗剂、滴剂或血清制剂。胃肠外给药制剂的非限制性实例包括但不限于注射用溶液剂、注射用干粉剂、注射用悬浮液、注射用乳剂等。本发明的药物组合物还可以制成控制释放或延迟释放剂型(例如脂质体或微球)。In the present invention, the purpose of using the pharmaceutical composition is to promote the administration to the living body, facilitate the absorption of the active ingredient, and then exert biological activity. The pharmaceutical compositions of the present invention can be administered in any form, including injection (intraarterial, intravenous, intramuscular, intraperitoneal, subcutaneous), mucosal, oral (oral solid, oral liquid), rectal, inhalation, implant , topical (eg ocular) administration, etc. Non-limiting examples of oral solid formulations include, but are not limited to, powders, capsules, lozenges, granules, tablets, and the like. Non-limiting examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, tinctures, elixirs, solutions, and the like. Non-limiting examples of formulations for topical administration include, but are not limited to, creams, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum formulations. Non-limiting examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry powders for injection, suspensions for injection, emulsions for injection, and the like. The pharmaceutical compositions of the present invention can also be formulated in controlled release or delayed release dosage forms (eg, liposomes or microspheres).

优选地，本发明中的化合物或包含其的药物组合物以口服或静脉内给药的方式施用于对其有需要的个体。取决于给药对象的具体情况，也可以应用甚至优选其它施用途经。例如，对于健忘或对口服药物易发怒的患者，经皮施用将是非常重要的给药方式。在本发明中，施用途经能够以任何适用的方式进行变化或调整，以满足药物的性质、患者和医务人员的便利以及其它相关因素的需求。Preferably, a compound of the present invention or a pharmaceutical composition comprising the same is administered orally or intravenously to an individual in need thereof. Depending on the particular circumstances of the subject being administered, other modes of administration may also be employed or even preferred. For example, for patients who are forgetful or irritable with oral medications, transdermal administration would be a very important mode of administration. In the present invention, the administration channel can be varied or adjusted in any suitable manner to meet the needs of the nature of the drug, the convenience of the patient and medical staff, and other relevant factors.

本发明的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药或者包含其的药物组合物具有优良的SHP2酶活性及细胞增殖抑制活性，能够作为SHP2抑制剂，用于预防和/或治疗与SHP2活性异常相关的(或者由SHP2异常突变引起的)疾病或病症，具有良好的临床应用和医药用途。优选地，与SHP2活性异常相关的(或者由SHP2异常突变引起的)疾病或病症的非限制性实例包括但不限于努南综合征、豹皮综合征、白血病(例如青少年髓单核细胞白血病、急性髓性白血病)、成神经细胞瘤、黑色素瘤、乳腺癌、食道癌、肺癌、结肠癌、头颈部肿瘤、胃癌、间变性大细胞淋巴瘤、成胶质细胞瘤等，优选非小细胞肺癌、食管癌和头颈部肿瘤。The compounds of the present invention or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, metabolites or prodrugs or pharmaceutical compositions containing them have excellent SHP2 enzyme activity and cellular The proliferation inhibitory activity can be used as a SHP2 inhibitor to prevent and/or treat diseases or conditions associated with abnormal SHP2 activity (or caused by abnormal SHP2 mutation), and has good clinical and medical applications. Preferably, non-limiting examples of diseases or conditions associated with abnormal SHP2 activity (or caused by abnormal SHP2 mutations) include, but are not limited to, Noonan syndrome, Leopard skin syndrome, leukemias (eg juvenile myelomonocytic leukemia, acute myeloid leukemia), neuroblastoma, melanoma, breast cancer, esophageal cancer, lung cancer, colon cancer, head and neck tumor, gastric cancer, anaplastic large cell lymphoma, glioblastoma, etc., preferably non-small cell Lung, esophageal and head and neck tumors.

以下将结合具体实施例来阐述本发明的技术方案，下列实施例的提供旨在进一步说明本发明，而非用于限制本发明的范围。对本领域技术人员而言，在不脱离本发明的精神和范围的情况下，针对本发明的具体实施方式进行各种变化和改进将是显而易见的。The technical solutions of the present invention will be described below with reference to specific embodiments. The following embodiments are provided to further illustrate the present invention, but not to limit the scope of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made in the specific embodiments of the present invention without departing from the spirit and scope of the invention.

本发明的化合物的制备可以通过本领域技术人员所熟知的合成方法来实现，包括但不限于下面列举的具体实施方式、其与其他化学合成方法相结合而形成的实施方式以及本领域技术人员所熟知的等同替换方式，优选的实施方式包括但不限于本发明的实施例。本发明中所使用的已知的起始原料可以提供本领域已知的方法来合成，或者通过常规的商业手段来购买(例如购自韶远化学科技、北京偶合科技等公司)。如无特殊说明，反应均在氩气氛或氮气氛下进行。氢化反应通常抽真空，充入氢气，反复操作3次。反应的温度为室温，温度范围是20℃-30℃。反应进程的监测可以通过本领域技术人员所熟知的合成方法来实现，包括但不限于薄层色谱法(TLC)。薄层层析硅胶板使用青岛海洋GF254硅胶板，展开剂体系包括但不限于A：二氯甲烷和甲醇体系；B：石油醚和乙酸乙酯体系，溶剂的体积比可以根据化合物的极性进行调节。The preparation of the compounds of the present invention can be achieved by synthetic methods well known to those skilled in the art, including but not limited to the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those skilled in the art. Well-known equivalents, preferred embodiments include, but are not limited to, the examples of the present invention. The known starting materials used in the present invention can be synthesized by methods known in the art, or purchased by conventional commercial means (for example, purchased from Shaoyuan Chemical Technology, Beijing Coupling Technology, etc.). Unless otherwise specified, the reactions were carried out in an argon atmosphere or a nitrogen atmosphere. The hydrogenation reaction is usually evacuated and filled with hydrogen, and the operation is repeated 3 times. The reaction temperature is room temperature, and the temperature range is 20°C-30°C. Monitoring the progress of the reaction can be accomplished by synthetic methods well known to those skilled in the art, including but not limited to thin layer chromatography (TLC). The thin layer chromatography silica gel plate uses Qingdao Ocean GF254 silica gel plate. The developing solvent system includes but is not limited to A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent can be determined according to the polarity of the compound. adjust.

本发明的化合物的分离纯化可以通过本领域技术人员所熟知的合成方法来实现，包括但不限于柱色谱法(CC)、高效液相色谱法(HPLC)、超高效液相色谱法(UPLC)等。柱色谱法一般使用青岛海洋200-300目硅胶作为载体，洗脱剂体系包括但不限于A：二氯甲烷和甲醇体系；B：石油醚和乙酸乙酯体系，溶剂的体积比可以根据化合物的极性进行调节，也可以加入少量的酸性或碱性防拖尾试剂进行调节。HPLC图谱采用Agilent 1200 DAD HPLC色谱仪(色谱柱：Sunfire C18,150×4.6mm,5μm)或Waters 2695-2996 HPLC色谱仪(色谱柱：Gimini C18,150×4.6mm,5μm)测定。The separation and purification of the compounds of the present invention can be achieved by synthetic methods well known to those skilled in the art, including but not limited to column chromatography (CC), high performance liquid chromatography (HPLC), ultra-high performance liquid chromatography (UPLC) Wait. Column chromatography generally uses Qingdao Ocean 200-300 mesh silica gel as the carrier, and the eluent system includes but is not limited to A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system, and the volume ratio of the solvent can be based on the compound. The polarity can be adjusted, and a small amount of acidic or basic anti-tailing reagents can also be added for adjustment. The HPLC chromatogram was determined using an Agilent 1200 DAD HPLC chromatograph (chromatographic column: Sunfire C18, 150×4.6mm, 5μm) or a Waters 2695-2996 HPLC chromatograph (chromatographic column: Gimini C18, 150×4.6mm, 5μm).

本发明的化合物的结构鉴定可以通过本领域技术人员所熟知的方法来实现，包括但不限于核磁共振(NMR)、质谱(MS)等。NMR图谱采用Bruker AVANCE-400或Varian Oxford-300核磁仪测定，测定溶剂为氘代二甲基亚砜(DMSO-d ₆)、氘代氯仿(CDC1 ₃)或氘代甲醇(CD ₃OD)，内标为四甲基硅烷(TMS)，化学位移以10 ^-6(ppm)计。MS 图谱采用Agilent SQD(ESI)质谱仪(型号：6110)或Shimadzu SQD(ESI)质谱仪(型号：2020)测定。 Structural identification of the compounds of the present invention can be accomplished by methods well known to those skilled in the art, including but not limited to nuclear magnetic resonance (NMR), mass spectrometry (MS), and the like. The NMR spectrum was determined by Bruker AVANCE-400 or Varian Oxford-300 nuclear magnetic instrument, and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDC1 ₃ ) or deuterated methanol (CD ₃ OD), The internal standard is tetramethylsilane (TMS), and the chemical shifts are in ^10-6 (ppm). MS spectra were determined using an Agilent SQD (ESI) mass spectrometer (model: 6110) or a Shimadzu SQD (ESI) mass spectrometer (model: 2020).

中间体的合成Synthesis of Intermediates

中间体Q1的合成Synthesis of Intermediate Q1

第一步：合成Q1-2Step 1: Synthesize Q1-2

将化合物Q1-1(100g,495mmol)加入到DME(1000ml)中，降温到-10℃后，加入氯甲酸异丁酯(67.6g,495mmol)和4-甲基吗啉(50g,495mmol)，室温反应5h，TLC监测反应结束后，过滤，用DME(250ml)过滤洗涤固体，然后将滤液加入到2L三口瓶中，加入硼氢化钠(37.6g,990mmol)处理滤液，室温下搅拌30min，再缓慢滴加甲醇(250ml)，继续室温反应3h，TLC监测反应完成，将反应液旋干后，加入水(1500ml)，水相用DCM萃取(300ml×3)，合并有机相后，有机相依次用水和饱和氯化钠溶液洗涤，再将有机相干燥，旋干后得到化合物Q1-2(86.5g,白色固体)，产率93％。粗品未经纯化，直接用于下一步反应。Compound Q1-1 (100g, 495mmol) was added to DME (1000ml), after cooling to -10°C, isobutyl chloroformate (67.6g, 495mmol) and 4-methylmorpholine (50g, 495mmol) were added, The reaction was carried out at room temperature for 5h, after the reaction was monitored by TLC, filtered, and the solid was filtered and washed with DME (250ml). Methanol (250ml) was slowly added dropwise, and the reaction was continued at room temperature for 3h. TLC monitored the completion of the reaction. After the reaction solution was spin-dried, water (1500ml) was added, and the aqueous phase was extracted with DCM (300ml×3). After combining the organic phases, the organic phases were sequentially Wash with water and saturated sodium chloride solution, then dry the organic phase, and spin dry to obtain compound Q1-2 (86.5 g, white solid) with a yield of 93%. The crude product was used directly in the next reaction without purification.

第二步：合成Q1-3Step 2: Synthesize Q1-3

将化合物Q1-2(25g,133mmol)加入到DCM(250ml)中，再加入TEA(26.9g,266mmol)，冰浴下滴加MsCl(18.3g,156mmol)，然后室温下反应1h。TLC显示反应结束后，将反应液用DCM稀释，依次用水和饱和氯化钠溶液洗涤，再将有机相干燥，浓缩旋干，残余物过硅胶层析柱(洗脱剂：V _石油醚:V _乙酸乙酯＝3:1～1:1)纯化，得到化合物Q1-3(33.6g,无色透明液体)，产率95％。 Compound Q1-2 (25 g, 133 mmol) was added to DCM (250 ml), then TEA (26.9 g, 266 mmol) was added, and MsCl (18.3 g, 156 mmol) was added dropwise under ice bath, and then reacted at room temperature for 1 h. After TLC showed that the reaction was completed, the reaction solution was diluted with DCM, washed with water and saturated sodium chloride solution in turn, then the organic phase was dried, concentrated and spun dry, and the residue was passed through a silica gel chromatography column (eluent: V _{petroleum ether} : V _{Ethyl acetate} =3:1～1:1) and purified to obtain compound Q1-3 (33.6 g, colorless transparent liquid) with a yield of 95%.

MS(ESI):m/z 266[M+H] ⁺。 MS(ESI): m/z 266[M+H] ⁺ .

第三步：合成Q1-4Step 3: Synthesize Q1-4

将N-叔丁氧羰基-4-氰基哌啶(28g,130mmol)加入到THF(300ml)中，冷却到-78℃后，缓慢滴加2.0M的LDA(75ml,150mmol)，滴加完成后，继续保持-78℃反应1.5h，然后滴加化合物Q1-3(26.6,100mmol)的THF溶液(150ml)，滴加完成后，继续保持 -78℃反应3h，TLC显示反应结束后，滴加饱和氯化铵溶液(50ml)淬灭反应，并加入饱和氯化钠溶液(500ml)，分出有机相，水相再用乙酸乙酯萃取(150ml×3)，合并有机相，再用无水硫酸钠干燥，过滤，旋干浓缩，残余物过硅胶层析柱(洗脱剂：V _石油醚:V _乙 _酸乙酯＝10:1～1:1)纯化，得到化合物Q1-4(25.7g,白色固体)，产率67.6％。 N-tert-butoxycarbonyl-4-cyanopiperidine (28g, 130mmol) was added to THF (300ml), cooled to -78°C, 2.0M LDA (75ml, 150mmol) was slowly added dropwise, and the dropwise addition was completed After the reaction was continued at -78°C for 1.5h, the THF solution (150ml) of compound Q1-3 (26.6, 100mmol) was added dropwise. After the addition was completed, the reaction was kept at -78°C for 3h. Saturated ammonium chloride solution (50ml) was added to quench the reaction, and saturated sodium chloride solution (500ml) was added, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (150ml×3), the organic phases were combined, Dry with water and sodium sulfate, filter, spin dry and concentrate, and the residue is purified by silica gel column chromatography (eluent: V _{petroleum ether} : V _{_{ethyl acetate}} =10:1～1:1) to obtain compound Q1-4 (25.7 g, white solid), 67.6% yield.

MS(ESI):m/z 380[M+H] ⁺。 MS (ESI): m/z 380 [M+H] ⁺ .

第四步：合成Q1-5Step 4: Synthesize Q1-5

将化合物Q1-4(25g,65.8mmol)加入到DMA(200ml)和水(20ml)的混合溶剂中，再加入三乙胺(33.2g,329mmol)和催化剂Pd(amphos)Cl ₂(4.6g,6.6mmol,CAS:887919-35-9)，氮气保护下于130℃反应4h。TLC显示反应结束后，将反应液冷却，并加入水稀释(800ml)，水相用乙酸乙酯萃取(200mL×3)，合并有机相，用饱和食盐水洗涤(200mL×2)，无水硫酸钠干燥，过滤，旋干浓缩，残余物过硅胶层析柱(洗脱剂：V _石油醚:V _乙酸乙酯＝3:1～1:1)纯化，得到化合物Q1-5(14.6g,白色固体)，产率73％。 Compound Q1-4 (25g, 65.8mmol) was added to a mixed solvent of DMA (200ml) and water (20ml), then triethylamine (33.2g, 329mmol) and catalyst Pd(amphos)Cl ₂ (4.6g, 6.6 mmol, CAS: 887919-35-9), reacted at 130° C. for 4 h under nitrogen protection. After TLC showed that the reaction was over, the reaction solution was cooled and diluted with water (800ml), the aqueous phase was extracted with ethyl acetate (200mL×3), the organic phases were combined, washed with saturated brine (200mL×2), anhydrous sulfuric acid Dry over sodium, filter, spin dry and concentrate, the residue is purified by silica gel column chromatography (eluent: V _{petroleum ether} : V _{ethyl acetate} =3:1～1:1) to obtain compound Q1-5 (14.6 g, white solid) in 73% yield.

MS(ESI):m/z 303[M+H] ⁺。 MS (ESI): m/z 303 [M+H] ⁺ .

第五步：合成Q1-6Step 5: Synthesis of Q1-6

将化合物Q1-5(10g,33mol)加入到钛酸四乙酯(100ml)中，然后再加入(R)-(+)-叔丁基亚磺酰胺(4.8g,40ml)，升温到90℃反应3h，TLC显示反应完成后，冷却到室温，将反应液缓慢加入到冰水(500ml)中，水相用二氯甲烷萃取(150mL×3)，合并有机相，用饱和食盐水洗涤(100mL×2)，无水硫酸钠干燥，过滤，旋干浓缩，残余物过硅胶层析柱(洗脱剂：V _石油醚:V _乙酸乙酯＝3:1～1:1)纯化，得到化合物Q1-6(12g,黄色固体)，产率89.6％。 Compound Q1-5 (10g, 33mol) was added to tetraethyl titanate (100ml), then (R)-(+)-tert-butylsulfinamide (4.8g, 40ml) was added, and the temperature was raised to 90°C After the reaction was completed for 3 h, TLC showed that the reaction was completed, cooled to room temperature, the reaction solution was slowly added to ice water (500 mL), the aqueous phase was extracted with dichloromethane (150 mL×3), the organic phases were combined, and washed with saturated brine (100 mL). ×2), dried over anhydrous sodium sulfate, filtered, concentrated by spin drying, and the residue was purified by silica gel column chromatography (eluent: V _{petroleum ether} : V _{ethyl acetate} =3:1～1:1) to obtain compound Q1 -6 (12 g, yellow solid) in 89.6% yield.

MS(ESI):m/z 406[M+H] ⁺。 MS(ESI): m/z 406[M+H] ⁺ .

第六步：合成Q1-7Step 6: Synthesis of Q1-7

将化合物Q1-6(10g,24.6mmol)加入到四氢呋喃(100ml)中，降温到-78℃后缓慢滴加DIBAL-H(30ml,30mmol,1M的甲苯溶液)，然后继续在-78℃反应0.5h，TLC显示反应完成后，在-50℃下加入饱和的罗谢尔盐溶液(300ml)淬灭反应，室温搅拌30min，水相用乙酸乙酯萃取(150mL×3)，合并有机相，用饱和食盐水洗涤(100mL×2)，无水硫酸钠干燥，过滤，旋干浓缩，残余物过硅胶层析柱(洗脱剂：V _石油醚:V _乙酸乙酯＝3:1～1:1)纯化，得到化合物Q1-7(8.6g,白色固体)，产率85.3％。 Compound Q1-6 (10g, 24.6mmol) was added to tetrahydrofuran (100ml), cooled to -78°C and slowly added dropwise DIBAL-H (30ml, 30mmol, 1M solution in toluene), and then continued to react at -78°C for 0.5 h, after TLC showed that the reaction was completed, saturated Rochelle salt solution (300ml) was added at -50°C to quench the reaction, stirred at room temperature for 30min, the aqueous phase was extracted with ethyl acetate (150mL×3), the organic phases were combined, and Washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, filtered, concentrated by spin drying, and the residue was passed through a silica gel chromatography column (eluent: V _{petroleum ether} : V _{ethyl acetate} =3:1～1:1 ) was purified to give compound Q1-7 (8.6 g, white solid) in 85.3% yield.

MS(ESI):m/z 408[M+H] ⁺。 MS(ESI): m/z 408 [M+H] ⁺ .

第七步：合成中间体Q1Step 7: Synthesis of Intermediate Q1

将化合物Q1-7(5g,12.2mmol)加入到乙酸乙酯(50ml)中，再加入4M的HCl/乙酸乙酯溶液(25ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到中间体Q1的盐酸盐(3.2g,白色固体)，产率94％。Compound Q1-7 (5 g, 12.2 mmol) was added to ethyl acetate (50 ml), then 4M HCl/ethyl acetate solution (25 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, it was filtered, and the solid was washed with acetic acid. Ethyl ester was washed and dried to obtain the hydrochloride salt of intermediate Q1 (3.2 g, white solid) in 94% yield.

MS(ESI):m/z 204[M+H] ⁺。 MS(ESI): m/z 204[M+H] ⁺ .

中间体Q2的合成Synthesis of Intermediate Q2

第一步：合成Q2-2Step 1: Synthesize Q2-2

将化合物Q2-1(205mg,1mmol)溶于二氧六环(5ml)中，然后加入3-巯基丙酸甲酯(180mg,1.5mmol)、Pd ₂(dba) ₃(22.73mg,0.025mmol,0.05当量)、Xantphos(14.36mg,0.025mmol)和DIEA(387mg,3mmol)。氮气保护下升温到90℃并搅拌1小时后，将反应混合物旋干浓缩，残余物过硅胶层析柱(洗脱剂：V _石油醚:V _乙酸乙酯＝3:1～1:1)纯化，得到化合物Q2-2(201mg,淡黄色液体)，83％产率。 Compound Q2-1 (205mg, 1mmol) was dissolved in dioxane (5ml), and then added methyl 3-mercaptopropionate (180mg, 1.5mmol), Pd ₂ (dba) ₃ (22.73mg, 0.025mmol, 0.05 equiv), Xantphos (14.36 mg, 0.025 mmol) and DIEA (387 mg, 3 mmol). Under nitrogen protection, the temperature was raised to 90°C and stirred for 1 hour, then the reaction mixture was spun dry and concentrated, and the residue was purified by silica gel column chromatography (eluent: V _{petroleum ether} : V _{ethyl acetate} =3:1～1:1) , to obtain compound Q2-2 (201 mg, pale yellow liquid) in 83% yield.

MS(ESI):m/z 246[M+H] ⁺。 MS(ESI): m/z 246[M+H] ⁺ .

第二步：合成中间体Q2Step 2: Synthesis of Intermediate Q2

将化合物Q2-2(200mg,0.81mmol)溶于THF(2ml)中，然后冷却到0℃，再加入叔丁醇钠(96mg,1mmol)，在0℃下搅拌0.5h，TLC显示反应结束后，将反应混合物用PE稀释，析出大量固体，通过过滤收集沉淀的固体，并用乙酸乙酯洗涤，得到中间体Q2(130mg,浅黄色固体)，产率88％。Compound Q2-2 (200 mg, 0.81 mmol) was dissolved in THF (2 ml), then cooled to 0 °C, then sodium tert-butoxide (96 mg, 1 mmol) was added, and stirred at 0 °C for 0.5 h, TLC showed that the reaction was over , the reaction mixture was diluted with PE, a large amount of solid was precipitated, the precipitated solid was collected by filtration, and washed with ethyl acetate to obtain intermediate Q2 (130 mg, pale yellow solid) in 88% yield.

MS(ESI):m/z 182[M+H] ⁺。 MS(ESI): m/z 182[M+H] ⁺ .

中间体Q3的合成Synthesis of Intermediate Q3

第一步：合成Q3-2Step 1: Synthesis of Q3-2

将化合物Q3-1(10g,106mmol)加入到二甲苯(100ml)中，然后加入甲烷三羧酸三乙酯(48g,222mmol,CAS:6279-86-3)，加完后升温到140℃反应3h，TLC显示反应结束后，冷却到室温，有固体析出，过滤，滤饼用***洗涤后，干燥，得到化合物Q3-2(16.2g,淡黄色固体)，65％产率。粗品未经纯化，直接用于下一步反应。Compound Q3-1 (10g, 106mmol) was added to xylene (100ml), then triethyl methanetricarboxylate (48g, 222mmol, CAS: 6279-86-3) was added, and the temperature was raised to 140°C after the addition was completed. After 3h, TLC showed that the reaction was completed, cooled to room temperature, a solid was precipitated, filtered, and the filter cake was washed with ether and dried to obtain compound Q3-2 (16.2 g, pale yellow solid) in 65% yield. The crude product was used directly in the next reaction without purification.

MS(ESI):m/z 235[M+H] ⁺。 MS (ESI): m/z 235 [M+H] ⁺ .

第二步：合成中间体Q3-3The second step: synthesis of intermediate Q3-3

将化合物Q3-2(5g,21mmol)加入到甲醇(40ml)和水(10ml)的混合溶剂中，套上氢气球，置换掉空气后，通氢气后反应3h，TLC显示反应结束后，过滤，滤饼用甲醇(50ml)洗涤，滤液旋干，得到化合物Q3-3(4.3g,淡黄色固体)，产率92％。粗品未经纯化，直接用于下一步反应。Compound Q3-2 (5g, 21mmol) was added to a mixed solvent of methanol (40ml) and water (10ml), covered with a hydrogen balloon, and after replacing the air, the reaction was carried out for 3h after passing through hydrogen. The filter cake was washed with methanol (50 ml), and the filtrate was spin-dried to obtain compound Q3-3 (4.3 g, pale yellow solid) with a yield of 92%. The crude product was used directly in the next reaction without purification.

MS(ESI):m/z 239[M+H] ⁺。 MS (ESI): m/z 239 [M+H] ⁺ .

第三步：合成中间体Q3The third step: synthesis of intermediate Q3

将化合物Q3-3(5g,21mmol)加入到甲醇(50ml)和水(10ml)的混合溶剂中，然后加入一水合氢氧化锂(1.7g,42mmol)，室温搅拌2h，TLC显示反应结束后，用1M盐酸调PH值到3-4，有大量固体析出，过滤，滤饼干燥，得到中间体Q3(3.98g,白色固体)，产率90％。粗品未经纯化，直接用于下一步反应。Compound Q3-3 (5g, 21mmol) was added to a mixed solvent of methanol (50ml) and water (10ml), then lithium hydroxide monohydrate (1.7g, 42mmol) was added, and stirred at room temperature for 2h. After TLC showed that the reaction was over, The pH value was adjusted to 3-4 with 1M hydrochloric acid, a large amount of solid was precipitated, filtered, and the filter cake was dried to obtain intermediate Q3 (3.98 g, white solid) with a yield of 90%. The crude product was used directly in the next reaction without purification.

MS(ESI):m/z 211[M+H] ⁺。 MS(ESI): m/z 211 [M+H] ⁺ .

中间体Q4的合成Synthesis of Intermediate Q4

第一步：合成Q4-1Step 1: Synthesis of Q4-1

将化合物Q2-2(1.5g，6.1mmol)和化合物Q3-3(1.2克，5.0mmol)在DMF(15毫升)中的溶液在160℃加热2.5小时。冷却后，将反应混合物加入到75ml饱和氯化钠溶液中，然用EtOAc萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，过滤并减压浓缩。用MeOH研磨浓缩物，得到化合物Q4-1(196g，黄色固体)，产率92％。A solution of compound Q2-2 (1.5 g, 6.1 mmol) and compound Q3-3 (1.2 g, 5.0 mmol) in DMF (15 mL) was heated at 160°C for 2.5 hours. After cooling, the reaction mixture was added to 75 ml of saturated sodium chloride solution, then extracted with EtOAc (25 ml×3), the organic phases were combined, washed with saturated sodium chloride solution (100 ml), dried over anhydrous sodium sulfate, filtered and reduced pressure concentrate. The concentrate was triturated with MeOH to give compound Q4-1 (196 g, yellow solid) in 92% yield.

MS(ESI):m/z 438.2[M+H] ⁺。 MS (ESI): m/z 438.2 [M+H] ⁺ .

第二步：合成Q4Step 2: Synthesize Q4

将化合物Q4-1(500mg,1.14mmol)溶于THF(2ml)中，然后冷却到0℃，再加入叔丁醇钠(131mg,1.37mmol)，在0℃下搅拌0.5h，TLC显示反应结束后，将反应混合物用PE稀释，析出大量固体，通过过滤收集沉淀的固体，并用乙酸乙酯洗涤，得到中间体Q4(370mg,浅黄色固体)，产率87％。Compound Q4-1 (500 mg, 1.14 mmol) was dissolved in THF (2 ml), then cooled to 0 °C, then sodium tert-butoxide (131 mg, 1.37 mmol) was added, stirred at 0 °C for 0.5 h, TLC showed that the reaction was complete After that, the reaction mixture was diluted with PE, a large amount of solid was precipitated, the precipitated solid was collected by filtration, and washed with ethyl acetate to obtain Intermediate Q4 (370 mg, pale yellow solid) in 87% yield.

中间体Q5的合成Synthesis of Intermediate Q5

第一步：合成Q5-2Step 1: Synthesis of Q5-2

将化合物2-氨基吡啶(4.35g，46.3mmol)和乙氧基亚甲基丙二酸二乙酯(10.00g，46.3mmol，cas:87-13-8)的混合物在120℃加热1小时。冷却到室温，再进一步冷却到5℃后，混合物用冰的乙醇进行打浆，过滤，固体用冰乙醇洗涤后干燥，得到化合物Q5-2(9.90g，黄色固体)，产率81％。A mixture of compound 2-aminopyridine (4.35 g, 46.3 mmol) and diethyl ethoxymethylenemalonate (10.00 g, 46.3 mmol, cas: 87-13-8) was heated at 120°C for 1 hour. After cooling to room temperature and further cooling to 5°C, the mixture was slurried with iced ethanol, filtered, and the solid was washed with iced ethanol and dried to obtain compound Q5-2 (9.90 g, yellow solid) in 81% yield.

MS(ESI):m/z 265.2[M+H] ⁺。 MS(ESI): m/z 265.2 [M+H] ⁺ .

第二步：合成Q5-3Step 2: Synthesis of Q5-3

将化合物Q5-2(13g，5.9mmol)加入到二苯醚(100ml)中，然后加热到280℃，回流2小时，TLC反应结束后，将反应混合物冷却至室温，并向其中加入正己烷(500ml)，以获得大量固体沉淀物。过滤沉淀的固体，用正己烷洗涤并干燥，得到化合物Q5-3(10g，黄色固体)，产率93.1％。Compound Q5-2 (13 g, 5.9 mmol) was added to diphenyl ether (100 ml), then heated to 280° C. and refluxed for 2 hours. After the TLC reaction was completed, the reaction mixture was cooled to room temperature, and n-hexane ( 500ml) to obtain a large amount of solid precipitate. The precipitated solid was filtered, washed with n-hexane and dried to obtain compound Q5-3 (10 g, yellow solid) in 93.1% yield.

MS(ESI):m/z 219.2[M+H] ⁺。 MS(ESI): m/z 219.2 [M+H] ⁺ .

第三步：合成Q5Step 3: Synthesize Q5

将化合物Q5-3(5g,23mmol)加入到甲醇(40ml)和水(10ml)的混合溶剂中，套上氢气球，置换掉空气后，通氢气后反应3h，TLC显示反应结束后，过滤，滤饼用甲醇(50ml)洗涤，滤液旋干，得到化合物Q5(4.6g,淡黄色固体)，产率92％。粗品无需纯化，直接用于下一步反应。Compound Q5-3 (5g, 23mmol) was added to a mixed solvent of methanol (40ml) and water (10ml), covered with a hydrogen balloon, and after replacing the air, the reaction was carried out for 3h after passing hydrogen. The filter cake was washed with methanol (50 ml), and the filtrate was spin-dried to obtain compound Q5 (4.6 g, pale yellow solid) in 92% yield. The crude product was directly used in the next reaction without purification.

中间体Q6的合成Synthesis of Intermediate Q6

第一步：合成Q6-2Step 1: Synthesis of Q6-2

MS(ESI):m/z 246[M+H] ⁺。 MS(ESI): m/z 246[M+H] ⁺ .

第二步：合成中间体Q6Step 2: Synthesis of Intermediate Q6

将化合物Q2-2(200mg,0.81mmol)溶于THF(2ml)中，然后冷却到0℃，再加入叔丁醇钠(96mg,1mmol)，在0℃下搅拌0.5h，TLC显示反应结束后，将反应混合物用PE稀释，析出大量固体，通过过滤收集沉淀的固体，并用乙酸乙酯洗涤，得到中间体Q2(130mg,浅黄色固体)，产率88％。Compound Q2-2 (200 mg, 0.81 mmol) was dissolved in THF (2 ml), then cooled to 0 °C, then sodium tert-butoxide (96 mg, 1 mmol) was added, and stirred at 0 °C for 0.5 h, TLC showed that the reaction was over. , the reaction mixture was diluted with PE, a large amount of solid was precipitated, the precipitated solid was collected by filtration, and washed with ethyl acetate to obtain Intermediate Q2 (130 mg, pale yellow solid) in 88% yield.

MS(ESI):m/z 182[M+H] ⁺。 MS(ESI): m/z 182[M+H] ⁺ .

中间体Q7的合成Synthesis of Intermediate Q7

第一步：合成Q7-2Step 1: Synthesis of Q7-2

MS(ESI):m/z 246[M+H] ⁺。 MS(ESI): m/z 246[M+H] ⁺ .

第二步：合成中间体Q7Step 2: Synthesis of Intermediate Q7

将化合物Q7-2(200mg,0.81mmol)溶于THF(2ml)中，然后冷却到0℃，再加入叔丁醇钠(96mg,1mmol)，在0℃下搅拌0.5h，TLC显示反应结束后，将反应混合物用PE稀释，析出大量固体，通过过滤收集沉淀的固体，并用乙酸乙酯洗涤，得到中间体Q2(130mg,浅黄色固体)，产率88％。Compound Q7-2 (200 mg, 0.81 mmol) was dissolved in THF (2 ml), then cooled to 0 °C, then sodium tert-butoxide (96 mg, 1 mmol) was added, and stirred at 0 °C for 0.5 h, TLC showed that the reaction was over , the reaction mixture was diluted with PE, a large amount of solid was precipitated, the precipitated solid was collected by filtration, and washed with ethyl acetate to obtain intermediate Q2 (130 mg, pale yellow solid) in 88% yield.

MS(ESI):m/z 182[M+H] ⁺。 MS(ESI): m/z 182[M+H] ⁺ .

实施例1：化合物1的制备Example 1: Preparation of Compound 1

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物1BStep 1: Synthesis of Compound 1B

将化合物1A(104mg,0.5mmol)溶于二氧六环(5ml)中，加入中间体Q2(182mg,1mmol)、Pd ₂(dba) ₃(22.73mg,0.025mmol)、Xantphos(14.36mg,0.025mmol)和DIEA(190mg,1.5mmol)。在氮气保护下升温到90℃并搅拌1h后，将反应混合物旋干浓缩，残余物过硅胶层析柱(洗脱剂：V _二氯甲烷:V _甲醇＝50:1～10:1)纯化，得到化合物1B(107mg,淡黄色固体)，75％产率。 Compound 1A (104 mg, 0.5 mmol) was dissolved in dioxane (5 ml), intermediate Q2 (182 mg, 1 mmol), Pd ₂ (dba) ₃ (22.73 mg, 0.025 mmol), Xantphos (14.36 mg, 0.025 were added) mmol) and DIEA (190 mg, 1.5 mmol). Under nitrogen protection, the temperature was raised to 90° C. and _stirred for 1 h, then the _reaction mixture was spun dry and concentrated. Compound 1B was obtained (107 mg, pale yellow solid) in 75% yield.

MS(ESI):m/z 287[M+H] ⁺。 MS(ESI): m/z 287[M+H] ⁺ .

第二步：合成化合物1CStep 2: Synthesis of Compound 1C

将化合物1B(100mg,0.35mmol)、DIEA(154mg,1.2mmol)、中间体Q3(147mg,0.7mmol)和HATU(456mg,1.2mmol)加入到DMF中，室温下搅拌12h，TLC显示反应结束后，反应液用二氯甲烷(10ml)稀释，有机相用饱和氯化钠水溶液洗涤，无水硫酸钠干燥，过滤，收集滤液，减压浓缩，残余物过硅胶层析柱(洗脱剂：V _二氯甲烷:V _甲醇＝50:1～10:1)纯化，得到化合物1B(137mg,淡黄色固体)，82％产率。 Compound 1B (100 mg, 0.35 mmol), DIEA (154 mg, 1.2 mmol), intermediate Q3 (147 mg, 0.7 mmol) and HATU (456 mg, 1.2 mmol) were added to DMF, and stirred at room temperature for 12 h. TLC showed that the reaction was over. , the reaction solution was diluted with dichloromethane (10 ml), the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, the filtrate was collected, concentrated under reduced pressure, and the residue was passed through a silica gel chromatography column (eluent: V _{Dichloromethane} :V _methanol =50:1～10:1) was purified to obtain compound 1B (137 mg, pale yellow solid) in 82% yield.

MS(ESI):m/z 479[M+H] ⁺。 MS (ESI): m/z 479 [M+H] ⁺ .

第三步：合成化合物1Step 3: Synthesis of Compound 1

将化合物1C(100mg,0.21mmol)、中间体Q1(71mg,0.35mmol)和DIEA(82mg,0.63mmol)溶于二甲基亚砜(5ml)中，90℃下反应1.5h。反应结束后，加入乙酸乙酯(20ml)和水(40ml)，水相用乙酸乙酯萃取(20ml×3)，合并有机相，用饱和氧化钠溶液洗涤(100ml)，无水硫酸钠干燥，过滤，收集滤液，减压浓缩，残余物过硅胶层析柱(洗脱剂：V _二氯甲烷:V _甲醇＝50:1～10:1)纯化，得到化合物1(32mg,淡黄色固体)。 Compound 1C (100 mg, 0.21 mmol), intermediate Q1 (71 mg, 0.35 mmol) and DIEA (82 mg, 0.63 mmol) were dissolved in dimethyl sulfoxide (5 ml) and reacted at 90° C. for 1.5 h. After the reaction, ethyl acetate (20ml) and water (40ml) were added, the aqueous phase was extracted with ethyl acetate (20ml×3), the organic phases were combined, washed with saturated sodium oxide solution (100ml), dried over anhydrous sodium sulfate, Filtration, the filtrate was collected, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: V _{dichloromethane} : V _methanol =50:1-10:1) to obtain compound 1 (32 mg, pale yellow solid).

MS(ESI):m/z 646[M+H] ⁺。 MS(ESI): m/z 646[M+H] ⁺ .

¹H-NMR(400MHz,DMSO-d ₆):δ12.26(brs,1H),8.54-8.53(m,1H),8.31(m,3H),8.15(m,1H),7.90-7.88(m,1H),7.70(s,1H),7.36-7.21(m,1H),6.47-6.45(m,1H),6.20-6.18(m,2H),4.48-4.67(m,1H),4.35-4.23(m,2H),3.87-3.67(m,2H),3.26-3.13(m,4H),2.90-2.87(m,2H),2.02-1.72(m,5H),1.55-1.45(m,3H)。 ¹ H-NMR (400MHz, DMSO-d ₆ ): δ12.26(brs, 1H), 8.54-8.53(m, 1H), 8.31(m, 3H), 8.15(m, 1H), 7.90-7.88(m ,1H),7.70(s,1H),7.36-7.21(m,1H),6.47-6.45(m,1H),6.20-6.18(m,2H),4.48-4.67(m,1H),4.35-4.23 (m,2H),3.87-3.67(m,2H),3.26-3.13(m,4H),2.90-2.87(m,2H),2.02-1.72(m,5H),1.55-1.45(m,3H) .

实施例2：化合物2的制备Example 2: Preparation of Compound 2

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物2BStep 1: Synthesis of Compound 2B

将化合物2A(1.0g,3.6mmol)、化合物Q1(808mg,4.0mmol)和DIEA(4.6g，36mmol)加入到MeCN(2240mL)中，然后将得到的溶液在80℃下搅拌2小时。TLC显示反应完成。将混合物冷却至25℃，并将Boc ₂O(1.6g，7.2mmol)加入溶液中。将反应在50℃下搅拌2小时。TLC显示反应完成。在减压下浓缩混合物。残余物过硅胶层析柱(洗脱剂:石油醚/乙酸乙酯＝1/2,V/V)纯化，得到化合物2B(953mg,淡黄色固体)，48.4％产率。 Compound 2A (1.0 g, 3.6 mmol), compound Q1 (808 mg, 4.0 mmol) and DIEA (4.6 g, 36 mmol) were added to MeCN (2240 mL), and the resulting solution was stirred at 80° C. for 2 hours. TLC showed the reaction was complete. The mixture was cooled to 25°C and Boc2O (1.6 _g , 7.2 mmol) was added to the solution. The reaction was stirred at 50°C for 2 hours. TLC showed the reaction was complete. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/2, V/V) to obtain compound 2B (953 mg, pale yellow solid) in 48.4% yield.

MS(ESI):m/z 547.1[M+H] ⁺。 MS (ESI): m/z 547.1 [M+H] ⁺ .

第二步：合成化合物2CStep 2: Synthesis of Compound 2C

将化合物2C(900mg,1.6mmol)溶于NMP(10ml)中，加入中间体Q4(895mg,2.4mmol)、Pd ₂(dba) ₃(145mg,0.16mmol)、Xantphos(92mg,0.16mmol)和DIEA(619mg,4.8mmol)。在氮气保护下升温到120℃并搅拌1h后，TLC反应结束后，将反应混合物加入到75ml饱和氯化钠溶液中，然用EtOAc萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水)，B:乙腈)；流速：25ml/min)，得到化合物2C(553mg,淡黄色固体)，45％产率。 Compound 2C (900 mg, 1.6 mmol) was dissolved in NMP (10 ml), intermediate Q4 (895 mg, 2.4 mmol), Pd2(dba ₎₃ ₍ 145 mg, 0.16 mmol), Xantphos (92 mg, 0.16 mmol) and DIEA were added (619 mg, 4.8 mmol). Under nitrogen protection, the temperature was raised to 120 °C and stirred for 1 h. After the TLC reaction was completed, the reaction mixture was added to 75 ml of saturated sodium chloride solution, then extracted with EtOAc (25 ml × 3), the organic phases were combined, and the saturated sodium chloride solution was used. The solution was washed (100ml), dried over anhydrous sodium sulfate, concentrated by spin drying, and the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05 % ammonia water), B: acetonitrile); flow rate: 25 ml/min) to give compound 2C (553 mg, pale yellow solid) in 45% yield.

MS(ESI):m/z 770.2[M+H] ⁺。 MS (ESI): m/z 770.2 [M+H] ⁺ .

第三步：合成化合物2Step 3: Synthesis of Compound 2

将化合物2C(500mg,0.65mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到化合物2的盐酸盐(400mg,白色固体)，产率94％。Compound 2C (500 mg, 0.65 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, it was filtered, and the solid was washed with ethyl acetate. Washed and dried to obtain the hydrochloride salt of compound 2 (400 mg, white solid) in 94% yield.

MS(ESI):m/z 670.2[M+H] ⁺。 MS (ESI): m/z 670.2 [M+H] ⁺ .

¹H NMR(400MHz,DMSO)δ12.28(s,1H),9.13-8.86(m,3H),8.67(d,J＝5.6Hz,1H),8.43-8.27(m,2H),8.22-8.16(m,1H),8.07(d,J＝25.2Hz,2H),7.66-7.57(m,1H),7.21(t,J＝8.0Hz,1H),6.71(d,J＝8.0Hz,1H),4.62-4.58(m,1H),4.16-4.13(m,1H),4.07-4.02(m,1H),3.88(t,J＝6.0Hz,2H),3.55-3.48(m,3H),3.30-3.24(m,1H),2.90(t,J＝6.4Hz,2H),2.18-2.09(m,1H),2.08-1.99(m,1H),1.95-1.88(m,2H),1.87-1.77(m,3H),1.70(d,J＝13.6Hz,1H)。 ¹ H NMR (400MHz, DMSO) δ 12.28(s, 1H), 9.13-8.86(m, 3H), 8.67(d, J=5.6Hz, 1H), 8.43-8.27(m, 2H), 8.22-8.16 (m,1H),8.07(d,J=25.2Hz,2H),7.66-7.57(m,1H),7.21(t,J=8.0Hz,1H),6.71(d,J=8.0Hz,1H) ,4.62-4.58(m,1H),4.16-4.13(m,1H),4.07-4.02(m,1H),3.88(t,J＝6.0Hz,2H),3.55-3.48(m,3H),3.30 -3.24(m, 1H), 2.90(t, J=6.4Hz, 2H), 2.18-2.09(m, 1H), 2.08-1.99(m, 1H), 1.95-1.88(m, 2H), 1.87-1.77 (m, 3H), 1.70 (d, J=13.6Hz, 1H).

实施例3：化合物3的制备Example 3: Preparation of Compound 3

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物3BStep 1: Synthesis of Compound 3B

将化合物3A(6g,24.80mmol)溶于DMF(50mL)中，在0℃下滴加DIEA(8.66mL,49.61mmol)和氨基乙醛缩二甲醇(2.90g，27.28mmol)的DMF溶液，升至室温反应2h。TLC显示反应结束后，加入水中，并用乙酸乙酯萃取，盐水洗有机相，用无水硫酸钠干燥有机相，残余物用硅胶柱层析纯化(洗脱剂：PE:EA＝5:1(体积比))，得到化合物3B(7.4g，淡黄色固体)，产率91％。Compound 3A (6 g, 24.80 mmol) was dissolved in DMF (50 mL), and a solution of DIEA (8.66 mL, 49.61 mmol) and aminoacetaldehyde dimethyl acetal (2.90 g, 27.28 mmol) in DMF was added dropwise at 0 °C, liters to room temperature for 2h. After TLC showed that the reaction was over, water was added, and extracted with ethyl acetate, the organic phase was washed with brine, dried with anhydrous sodium sulfate, and the residue was purified by silica gel column chromatography (eluent: PE:EA=5:1 ( volume ratio)) to obtain compound 3B (7.4 g, pale yellow solid) in 91% yield.

MS(ESI):m/z310[M+H] ⁺。 MS(ESI): m/z310[M+H] ⁺ .

第二步：合成化合物3CStep 2: Synthesis of compound 3C

将化合物3B(5g,16.10mmol)在冰浴下分批缓慢加入浓硫酸(15mL)中，升温至75度反应2h。TLC显示反应结束，将反应液冷却至0℃，滴加5N氢氧化钠水溶液调pH为6，有黄色固体析出，过滤固体并烘干，得到化合物3C(2.77g，黄色固体)，产率71％。Compound 3B (5 g, 16.10 mmol) was slowly added to concentrated sulfuric acid (15 mL) in batches under an ice bath, and the temperature was raised to 75 degrees to react for 2 h. TLC showed that the reaction was completed, the reaction solution was cooled to 0°C, and 5N aqueous sodium hydroxide solution was added dropwise to adjust the pH to 6, a yellow solid was precipitated, the solid was filtered and dried to obtain compound 3C (2.77g, yellow solid) in a yield of 71 %.

MS(ESI):m/z 228[M+H] ⁺。 MS(ESI): m/z 228[M+H] ⁺ .

第三步：合成化合物3DStep 3: Synthesize Compounds 3D

将化合物3C(2.77g,12.15mmol)加入到POCl ₃(30mL)中，在0℃下滴加DIEA(4.24mL,24.29mmol)，升温至115度反应15min。TLC显示反应结束后，旋干反应液，加入乙酸乙酯并缓慢倒入水中，用饱和碳酸氢钠水溶液调pH为中性，萃取，盐水洗有机相，用无水硫酸钠干燥，残余物用硅胶柱层析纯化(洗脱剂，PE:EA＝3:1(体积比))，得到化合物3D(1.17g，淡黄色固体)，产率38％。 Compound 3C (2.77 g, 12.15 mmol) was added to POCl ₃ (30 mL), DIEA (4.24 mL, 24.29 mmol) was added dropwise at 0° C., the temperature was raised to 115 degrees and the reaction was performed for 15 min. After TLC showed that the reaction was completed, the reaction solution was spin-dried, ethyl acetate was added and slowly poured into water, the pH was adjusted to neutral with saturated aqueous sodium bicarbonate solution, extracted, and the organic phase was washed with brine, dried over anhydrous sodium sulfate, and the residue was treated with Purification by silica gel column chromatography (eluent, PE:EA=3:1 (volume ratio)) gave compound 3D (1.17 g, pale yellow solid) in 38% yield.

MS(ESI):m/z 246[M+H] ⁺。 MS(ESI): m/z 246[M+H] ⁺ .

第四步：合成化合物3EStep 4: Synthesis of Compound 3E

将化合物3D(886mg,3.6mmol)、化合物Q1(808mg,4.0mmol)和DIEA(4.6g，36mmol)加入到MeCN(40mL)中，然后将得到的溶液在80℃下搅拌2小时。TLC显示反应完成。将混合物冷却至25℃，并将Boc ₂O(1.6g，7.2mmol)加入溶液中。将反应在50℃下搅拌2小时。TLC显示反应完成。在减压下浓缩混合物。残余物过硅胶层析柱(洗脱剂:石油醚/乙酸乙酯＝1/2,V/V)纯化，得到化合物3E(786mg,淡黄色固体)，38.9％产率。 Compound 3D (886 mg, 3.6 mmol), compound Q1 (808 mg, 4.0 mmol) and DIEA (4.6 g, 36 mmol) were added to MeCN (40 mL), and the resulting solution was stirred at 80°C for 2 hours. TLC showed the reaction was complete. The mixture was cooled to 25°C and Boc2O (1.6 _g , 7.2 mmol) was added to the solution. The reaction was stirred at 50°C for 2 hours. TLC showed the reaction was complete. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/2, V/V) to obtain compound 3E (786 mg, pale yellow solid) in 38.9% yield.

MS(ESI):m/z 561.1[M+H] ⁺。 MS(ESI): m/z 561.1 [M+H] ⁺ .

第五步：合成化合物3FStep 5: Synthesis of Compound 3F

将化合物3E(700mg,1.24mmol)溶于NMP(10ml)中，加入中间体Q4(895mg,2.4mmol)、Pd ₂(dba) ₃(145mg,0.16mmol)、Xantphos(92mg,0.16mmol)和DIEA(619mg,4.8mmol)。在氮气保护下升温到120℃并搅拌1h后，TLC反应结束后，将反应混合物加入到75ml饱和氯化钠溶液中，然用EtOAc萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物3F(340mg,淡黄色固体)，35％产率。 Compound 3E (700 mg, 1.24 mmol) was dissolved in NMP (10 ml), intermediate Q4 (895 mg, 2.4 mmol), Pd2(dba ₎₃ ₍ 145 mg, 0.16 mmol), Xantphos (92 mg, 0.16 mmol) and DIEA were added (619 mg, 4.8 mmol). Under nitrogen protection, the temperature was raised to 120 °C and stirred for 1 h. After the TLC reaction was completed, the reaction mixture was added to 75 ml of saturated sodium chloride solution, then extracted with EtOAc (25 ml × 3), the organic phases were combined, and the saturated sodium chloride solution was used. The solution was washed (100ml), dried over anhydrous sodium sulfate, concentrated by spin drying, and the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05 % ammonia water), B: acetonitrile); flow rate: 25 ml/min) to give compound 3F (340 mg, pale yellow solid) in 35% yield.

MS(ESI):m/z 784.2[M+H] ⁺。 MS (ESI): m/z 784.2 [M+H] ⁺ .

第六步：合成化合物3Step 6: Synthesis of Compound 3

将化合物3F(300mg,0.38mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到化合物3的盐酸盐(239mg,白色固体)，产率92％。Compound 3F (300 mg, 0.38 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, it was filtered, and the solid was washed with ethyl acetate. Washed and dried to obtain the hydrochloride salt of compound 3 (239 mg, white solid) in 92% yield.

MS(ESI):m/z 684.2[M+H] ⁺。 MS(ESI): m/z 684.2 [M+H] ⁺ .

¹H NMR(400MHz,DMSO)δ12.28(s,1H),9.31(s,1H),8.55(d,J＝4.8Hz,1H),8.49-8.33(m,2H),8.14(d,J＝8.4Hz,1H),7.93(d,J＝7.2Hz,1H),7.35(dd,J＝7.6,4.8Hz,1H),7.28-6.96(m,3H),6.54(d,J＝8.4Hz,1H),4.53(d,J＝4.8Hz,1H),4.23(d,J＝13.6Hz,1H),4.14(d,J＝14.4Hz,1H),3.89(t,J＝5.6Hz,2H),3.60-3.54(m,2H),3.28(d,J＝16.8Hz,1H),3.14(d,J＝16.8Hz,1H),2.89(t,J＝6.5Hz,2H),2.52(s,3H),2.05-1.87(m,3H),1.86-1.69(m,3H),1.62(d,J＝12.8Hz,1H),1.28-1.21(m,1H)。 ¹ H NMR (400MHz, DMSO) δ 12.28(s, 1H), 9.31(s, 1H), 8.55(d, J=4.8Hz, 1H), 8.49-8.33(m, 2H), 8.14(d, J ＝8.4Hz,1H),7.93(d,J＝7.2Hz,1H),7.35(dd,J＝7.6,4.8Hz,1H),7.28-6.96(m,3H),6.54(d,J＝8.4Hz ,1H),4.53(d,J＝4.8Hz,1H),4.23(d,J＝13.6Hz,1H),4.14(d,J＝14.4Hz,1H),3.89(t,J＝5.6Hz,2H ),3.60-3.54(m,2H),3.28(d,J＝16.8Hz,1H),3.14(d,J＝16.8Hz,1H),2.89(t,J＝6.5Hz,2H),2.52(s , 3H), 2.05-1.87 (m, 3H), 1.86-1.69 (m, 3H), 1.62 (d, J=12.8Hz, 1H), 1.28-1.21 (m, 1H).

实施例4：化合物4的制备Example 4: Preparation of Compound 4

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物4BStep 1: Synthesis of Compound 4B

将化合物4A(10g,71mmol)溶于THF(100ml)中，室温下分批缓慢加入NIS(17.6g，78mmol)，室温反应过夜。LCMS检测反应完全，将反应液加入到500ml水中，析出大量固体，过滤得到的固体再用二氯甲烷洗涤，干燥得到化合物4B(16.5g,淡黄色固体)，87％收率。Compound 4A (10 g, 71 mmol) was dissolved in THF (100 ml), NIS (17.6 g, 78 mmol) was slowly added in portions at room temperature, and the reaction was carried out at room temperature overnight. LCMS detected that the reaction was complete, the reaction solution was added to 500 ml of water, a large amount of solid was precipitated, the solid obtained by filtration was washed with dichloromethane, and dried to obtain compound 4B (16.5 g, pale yellow solid) in 87% yield.

MS(ESI):m/z 268.1[M+H] ⁺。 MS(ESI): m/z 268.1 [M+H] ⁺ .

第二步：合成化合物4CStep 2: Synthesis of compound 4C

将化合物Q1(1.0g,4.9mmol)加入到DMF(10ml)中，然后加入化合物4B(2.62g,9.8mmol)和BOP(4.3g,9.8mmol)，加完后室温下缓慢加入DBU(2.2g,14.7mmol)，然后室温反应过夜，反应结束后，将Boc ₂O(1.6g，7.2mmol)加入溶液中。将反应在50℃下搅拌2小时。TLC显示反应完成。在减压下浓缩混合物。残余物过硅胶层析柱(洗脱剂:石油醚/乙酸乙酯＝1/2,V/V)纯化，得到化合物4C(1.1g,淡黄色固体)，42.3％产率。 Compound Q1 (1.0 g, 4.9 mmol) was added to DMF (10 ml), then compound 4B (2.62 g, 9.8 mmol) and BOP (4.3 g, 9.8 mmol) were added, and DBU (2.2 g) was slowly added at room temperature after the addition. , 14.7 mmol), and then reacted at room temperature overnight. After the reaction was completed, Boc ₂ O (1.6 g, 7.2 mmol) was added to the solution. The reaction was stirred at 50°C for 2 hours. TLC showed the reaction was complete. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/2, V/V) to obtain compound 4C (1.1 g, pale yellow solid) in 42.3% yield.

MS(ESI):m/z 553.1[M+H] ⁺。 MS (ESI): m/z 553.1 [M+H] ⁺ .

第三步：合成化合物4DStep 3: Synthesis of Compound 4D

将化合物4C(830mg,1.5mmol)溶于NMP(10ml)中，加入中间体Q4(895mg,2.4mmol)、Pd ₂(dba) ₃(145mg,0.16mmol)、Xantphos(92mg,0.16mmol)和DIEA(619mg,4.8mmol)。在氮气保护下升温到120℃并搅拌1h后，TLC反应结束后，将反应混合物加入到75ml饱和氯化钠溶液中，然用EtOAc萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物4D(384mg,淡黄色固体)，33％产率。 Compound 4C (830 mg, 1.5 mmol) was dissolved in NMP (10 ml), intermediate Q4 (895 mg, 2.4 mmol), Pd2(dba ₎₃ ₍ 145 mg, 0.16 mmol), Xantphos (92 mg, 0.16 mmol) and DIEA were added (619 mg, 4.8 mmol). Under nitrogen protection, the temperature was raised to 120 °C and stirred for 1 h. After the TLC reaction was completed, the reaction mixture was added to 75 ml of saturated sodium chloride solution, then extracted with EtOAc (25 ml × 3), the organic phases were combined, and the saturated sodium chloride solution was used. The solution was washed (100ml), dried over anhydrous sodium sulfate, concentrated by spin drying, and the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05 % ammonia water), B: acetonitrile); flow rate: 25 ml/min) to give compound 4D (384 mg, pale yellow solid) in 33% yield.

MS(ESI):m/z 776.2[M+H] ⁺。 MS (ESI): m/z 776.2 [M+H] ⁺ .

第四步：合成化合物4Step 4: Synthesis of Compound 4

将化合物4D(350mg,0.45mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到化合物4的盐酸盐(276mg,白色固体)，产率91％。Compound 4D (350 mg, 0.45 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was complete, it was filtered, and the solid was washed with ethyl acetate. Washed and dried to obtain the hydrochloride salt of compound 4 (276 mg, white solid) in 91% yield.

MS(ESI):m/z 676.2[M+H] ⁺。 MS(ESI): m/z 676.2 [M+H] ⁺ .

实施例5：化合物5的制备Example 5: Preparation of Compound 5

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物5AStep 1: Synthesis of Compound 5A

将化合物1B(500mg,1.75mmol)和化合物Q5(388mg,1.75mmol)加入到NMP(10ml)中，然后在氮气保护下升温到120℃并搅拌1h后，TLC反应结束后，将反应混合物加入到75ml饱和氯化钠溶液中，然用EtOAc萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用硅胶柱层析纯化(洗脱剂，PE:EA＝3:1(体积比))，得到化合物5A(339mg，淡黄色固体)，产率42％。Compound 1B (500 mg, 1.75 mmol) and compound Q5 (388 mg, 1.75 mmol) were added to NMP (10 ml), and then heated to 120° C. under nitrogen protection and stirred for 1 h. After the TLC reaction was completed, the reaction mixture was added to 75ml of saturated sodium chloride solution, then extracted with EtOAc (25ml × 3), the organic phases were combined, washed with saturated sodium chloride solution (100ml), dried over anhydrous sodium sulfate, spin-dried and concentrated, and the residue was subjected to silica gel column chromatography Purification (eluent, PE:EA=3:1 (v/v)) afforded compound 5A (339 mg, pale yellow solid) in 42% yield.

MS(ESI):m/z 463.2[M+H] ⁺。 MS(ESI): m/z 463.2 [M+H] ⁺ .

第三步：合成化合物5Step 3: Synthesis of Compound 5

将化合物5A(300mg,0.64mmol)、化合物Q1(156mg,0.77mmol)和DIEA(826mg，6.4mmol)加入到MeCN(10mL)中，然后将得到的溶液在80℃下搅拌2小时。TLC显示反应完成。在减压下浓缩混合物。残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物5(106mg,淡黄色固体)，26.2％产率。 Compound 5A (300 mg, 0.64 mmol), compound Q1 (156 mg, 0.77 mmol) and DIEA (826 mg, 6.4 mmol) were added to MeCN (10 mL), and the resulting solution was stirred at 80°C for 2 hours. TLC showed the reaction was complete. The mixture was concentrated under reduced pressure. The residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05% ammonia), B: acetonitrile); flow rate: 25ml/min), Compound 5 (106 mg, pale yellow solid) was obtained in 26.2% yield.

MS(ESI):m/z 630.2[M+H] ⁺。 MS(ESI): m/z 630.2 [M+H] ⁺ .

¹H NMR(400MHz,DMSO)δ11.28(s,1H),8.53(d,J＝4.2Hz,1H),8.43-8.30(m,2H),7.94-7.85(m,1H),7.72-7.63(m,1H),7.54-7.45(m,1H),7.37-7.29(m,1H),7.26-7.10(m,2H),6.55-6.43(m,1H),6.34-6.25(m,1H),6.13(s,1H),4.47(s,1H),4.28(dd,J＝34.4,12.4Hz,2H),3.46(s,1H),3.27-3.15(m,3H),3.15-3.00(m,2H),2.93-2.77(m,2H),2.05-1.83(m,2H),1.83-1.64(m,3H),1.62-1.46(m,2H),1.42-1.32(m,1H)。 ¹ H NMR (400MHz, DMSO) δ 11.28(s, 1H), 8.53(d, J=4.2Hz, 1H), 8.43-8.30(m, 2H), 7.94-7.85(m, 1H), 7.72-7.63 (m,1H),7.54-7.45(m,1H),7.37-7.29(m,1H),7.26-7.10(m,2H),6.55-6.43(m,1H),6.34-6.25(m,1H) ,6.13(s,1H),4.47(s,1H),4.28(dd,J=34.4,12.4Hz,2H),3.46(s,1H),3.27-3.15(m,3H),3.15-3.00(m , 2H), 2.93-2.77(m, 2H), 2.05-1.83(m, 2H), 1.83-1.64(m, 3H), 1.62-1.46(m, 2H), 1.42-1.32(m, 1H).

实施例6：化合物6的制备Example 6: Preparation of Compound 6

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物6AStep 1: Synthesis of Compound 6A

将化合物1A(207mg,1.0mmol)溶于二氧六环(5ml)中，加入中间体Q6(322mg,2mmol)、Pd ₂(dba) ₃(45mg,0.05mmol)、Xantphos(28mg,0.05mmol)和DIEA(380mg,3.0mmol)。在氮气保护下升温到90℃并搅拌1h后，将反应混合物旋干浓缩，残余物过硅胶层析柱(洗脱剂：V _二氯甲烷:V _甲醇＝50:1～10:1)纯化，得到化合物6A(173mg,淡黄色固体)，65％产率。 Compound 1A (207 mg, 1.0 mmol) was dissolved in dioxane (5 ml), intermediate Q6 (322 mg, 2 mmol), Pd ₂ (dba) ₃ (45 mg, 0.05 mmol), Xantphos (28 mg, 0.05 mmol) were added and DIEA (380 mg, 3.0 mmol). Under nitrogen protection, the temperature was raised to 90°C and stirred for 1 h. The reaction mixture was concentrated by spin drying, and the residue was purified by silica gel column chromatography (eluent: V _{dichloromethane} : V _methanol =50:1～10:1). Compound 6A (173 mg, pale yellow solid) was obtained in 65% yield.

MS(ESI):m/z 267.2[M+H] ⁺。 MS(ESI): m/z 267.2 [M+H] ⁺ .

第二步：合成化合物6BStep 2: Synthesis of Compound 6B

将化合物6A(100mg,0.37mmol)、DIEA(154mg,1.2mmol)、中间体Q3(147mg,0.7mmol)和HATU(456mg,1.2mmol)加入到DMF中，室温下搅拌12h，TLC显示反应结束后，反应液用二氯甲烷(10ml)稀释，有机相用饱和氯化钠水溶液洗涤，无水硫酸钠干燥，过滤，收集滤液，减压浓缩，残余物过硅胶层析柱(洗脱剂：V _二氯甲烷:V _甲醇＝50:1～10:1)纯化，得到化合物6B(138mg,淡黄色固体)，81.5％产率。 Compound 6A (100 mg, 0.37 mmol), DIEA (154 mg, 1.2 mmol), intermediate Q3 (147 mg, 0.7 mmol) and HATU (456 mg, 1.2 mmol) were added to DMF, and stirred at room temperature for 12 h. TLC showed that the reaction was over. , the reaction solution was diluted with dichloromethane (10 ml), the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, the filtrate was collected, concentrated under reduced pressure, and the residue was passed through a silica gel chromatography column (eluent: V _{Dichloromethane} :V _methanol =50:1～10:1) was purified to obtain compound 6B (138 mg, pale yellow solid) in 81.5% yield.

MS(ESI):m/z 459.2[M+H]MS(ESI): m/z 459.2[M+H]

第三步：合成化合物6Step 3: Synthesis of Compound 6

将化合物6B(100mg,0.22mmol)、中间体Q1(71mg,0.35mmol)和DIEA(82mg,0.63mmol)溶于二甲基亚砜(5ml)中，90℃下反应1.5h。反应结束后，加入乙酸乙酯(20ml)和水(40ml)，水相用乙酸乙酯萃取(20ml×3)，合并有机相，用饱和氧化钠溶液洗涤(100ml)，无水硫酸钠干燥，过滤，收集滤液，减压浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物6(30.7mg,淡黄色固体)，22.3％产率。 Compound 6B (100 mg, 0.22 mmol), intermediate Q1 (71 mg, 0.35 mmol) and DIEA (82 mg, 0.63 mmol) were dissolved in dimethyl sulfoxide (5 ml) and reacted at 90° C. for 1.5 h. After the reaction, ethyl acetate (20ml) and water (40ml) were added, the aqueous phase was extracted with ethyl acetate (20ml×3), the organic phases were combined, washed with saturated sodium oxide solution (100ml), dried over anhydrous sodium sulfate, Filtration, collecting the filtrate, and concentrating under reduced pressure, the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05% ammonia water), B: acetonitrile) ); flow rate: 25 ml/min) to give compound 6 (30.7 mg, pale yellow solid) in 22.3% yield.

MS(ESI):m/z 626.2[M+H] ⁺。 MS(ESI): m/z 626.2 [M+H] ⁺ .

实施例7：化合物7的制备Example 7: Preparation of Compound 7

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物7AStep 1: Synthesis of Compound 7A

将化合物1A(207mg,1.0mmol)溶于二氧六环(5ml)中，加入中间体Q7(358mg,2mmol)、Pd ₂(dba) ₃(45mg,0.05mmol)、Xantphos(28mg,0.05mmol)和DIEA(380mg,3.0mmol)。在氮气保护下升温到90℃并搅拌1h后，将反应混合物旋干浓缩，残余物过硅胶层析柱(洗脱剂,二氯甲烷:甲醇＝50:1～10:1，V：V)纯化，得到化合物7A(194mg,淡黄色固体)，63.5％产率。 Compound 1A (207 mg, 1.0 mmol) was dissolved in dioxane (5 ml), intermediate Q7 (358 mg, 2 mmol), Pd ₂ (dba) ₃ (45 mg, 0.05 mmol), Xantphos (28 mg, 0.05 mmol) were added and DIEA (380 mg, 3.0 mmol). Under nitrogen protection, the temperature was raised to 90°C and stirred for 1 h. The reaction mixture was concentrated by spin drying, and the residue was passed through a silica gel chromatography column (eluent, dichloromethane:methanol=50:1～10:1, V:V) Purification gave compound 7A (194 mg, pale yellow solid) in 63.5% yield.

MS(ESI):m/z 305.1[M+H] ⁺。 MS (ESI): m/z 305.1 [M+H] ⁺ .

第二步：合成化合物7BStep 2: Synthesis of Compound 7B

将化合物7A(106mg,0.35mmol)、DIEA(154mg,1.2mmol)、中间体Q3(147mg,0.7mmol)和HATU(456mg,1.2mmol)加入到DMF中，室温下搅拌12h，TLC显示反应结束后，反应液用二氯甲烷(10ml)稀释，有机相用饱和氯化钠水溶液洗涤，无水硫酸钠干燥，过滤，收集滤液，减压浓缩，残余物过硅胶层析柱(洗脱剂,二氯甲烷:甲醇＝50:1～10:1，V：V)纯化，得到化合物7B(145mg,淡黄色固体)，83.6％产率。Compound 7A (106 mg, 0.35 mmol), DIEA (154 mg, 1.2 mmol), intermediate Q3 (147 mg, 0.7 mmol) and HATU (456 mg, 1.2 mmol) were added to DMF, and stirred at room temperature for 12 h. TLC showed that the reaction was over. , the reaction solution was diluted with dichloromethane (10 ml), the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, the filtrate was collected, concentrated under reduced pressure, and the residue was subjected to silica gel chromatography (eluent, di Chloromethane:methanol=50:1～10:1, V:V) was purified to obtain compound 7B (145 mg, pale yellow solid) in 83.6% yield.

MS(ESI):m/z 497.2[M+H]MS(ESI): m/z 497.2[M+H]

第三步：合成化合物7Step 3: Synthesis of Compound 7

将化合物7B(110mg,0.22mmol)、中间体Q1(71mg,0.35mmol)和DIEA(82mg, 0.63mmol)溶于二甲基亚砜(5ml)中，90℃下反应1.5h。反应结束后，加入乙酸乙酯(20ml)和水(40ml)，水相用乙酸乙酯萃取(20ml×3)，合并有机相，用饱和氧化钠溶液洗涤(100ml)，无水硫酸钠干燥，过滤，收集滤液，减压浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物7(37.2mg,淡黄色固体)，26.2％产率。 Compound 7B (110 mg, 0.22 mmol), intermediate Q1 (71 mg, 0.35 mmol) and DIEA (82 mg, 0.63 mmol) were dissolved in dimethyl sulfoxide (5 ml) and reacted at 90° C. for 1.5 h. After the reaction, ethyl acetate (20ml) and water (40ml) were added, the aqueous phase was extracted with ethyl acetate (20ml×3), the organic phases were combined, washed with saturated sodium oxide solution (100ml), dried over anhydrous sodium sulfate, Filtration, collecting the filtrate, and concentrating under reduced pressure, the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05% ammonia water), B: acetonitrile) ); flow rate: 25 ml/min) to give compound 7 (37.2 mg, pale yellow solid) in 26.2% yield.

MS(ESI):m/z 644.2[M+H] ⁺。 MS(ESI): m/z 644.2 [M+H] ⁺ .

¹H NMR(400MHz,DMSO)δ12.87-12.69(m,1H),8.44(d,J＝4.4Hz,1H),8.03-7.93(m,1H),7.89-7.76(m,2H),7.69-7.62(m,2H),7.26(dd,J＝7.6,5.2Hz,1H),6.30-6.03(m,3H),4.24(t,J＝17.6Hz,2H),4.12(s,2H),3.82-3.68(m,2H),2.96(d,J＝16.8Hz,1H),2.72-2.62(m,2H),2.56-2.52(m,2H),2.36-2.22(m,3H),1.89-1.66(m,6H),1.54(t,J＝14.0Hz,1H),1.41(d,J＝12.4Hz,1H)。 ¹ H NMR (400MHz, DMSO) δ 12.87-12.69 (m, 1H), 8.44 (d, J=4.4Hz, 1H), 8.03-7.93 (m, 1H), 7.89-7.76 (m, 2H), 7.69 -7.62(m, 2H), 7.26(dd, J=7.6, 5.2Hz, 1H), 6.30-6.03(m, 3H), 4.24(t, J=17.6Hz, 2H), 4.12(s, 2H), 3.82-3.68(m, 2H), 2.96(d, J=16.8Hz, 1H), 2.72-2.62(m, 2H), 2.56-2.52(m, 2H), 2.36-2.22(m, 3H), 1.89- 1.66 (m, 6H), 1.54 (t, J=14.0 Hz, 1H), 1.41 (d, J=12.4 Hz, 1H).

实施例8：化合物8的制备Example 8: Preparation of Compound 8

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物8BStep 1: Synthesis of Compound 8B

将化合物Q1(1.0g,4.9mmol)加入到DMF(10ml)中，然后加入化合物8A(2.45g,9.8mmol)和BOP(4.3g,9.8mmol)，加完后室温下缓慢加入DBU(2.2g,14.7mmol)，然后室温反应过夜，反应结束后，将Boc ₂O(1.6g，7.2mmol)加入溶液中。将反应在50℃下搅拌2小时。TLC显示反应完成。在减压下浓缩混合物。残余物过硅胶层析柱(洗脱剂:石油醚/乙酸乙酯＝1/2,V/V)纯化，得到化合物8B(1.15g,淡黄色固体)，45.2％产率。 Compound Q1 (1.0 g, 4.9 mmol) was added to DMF (10 ml), then compound 8A (2.45 g, 9.8 mmol) and BOP (4.3 g, 9.8 mmol) were added, and DBU (2.2 g) was slowly added at room temperature after the addition. , 14.7 mmol), and then reacted at room temperature overnight. After the reaction was completed, Boc ₂ O (1.6 g, 7.2 mmol) was added to the solution. The reaction was stirred at 50°C for 2 hours. TLC showed the reaction was complete. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/2, V/V) to obtain compound 8B (1.15 g, pale yellow solid) in 45.2% yield.

MS(ESI):m/z 518.1[M+H] ⁺。 MS (ESI): m/z 518.1 [M+H] ⁺ .

第二步：合成化合物8CStep 2: Synthesis of Compound 8C

将化合物8B(829mg,1.6mmol)溶于NMP(10ml)中，加入中间体Q4(895mg,2.4mmol)、Pd ₂(dba) ₃(145mg,0.16mmol)、Xantphos(92mg,0.16mmol)和DIEA(619mg,4.8mmol)。在氮气保护下升温到120℃并搅拌1h后，TLC反应结束后，将反应混合物加入到75ml饱和氯化钠溶液中，然用EtOAc萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物8C(412mg,淡黄色固体)，32.6％产率。 Compound 8B (829 mg, 1.6 mmol) was dissolved in NMP (10 ml), intermediate Q4 (895 mg, 2.4 mmol), Pd2(dba ₎₃ ₍ 145 mg, 0.16 mmol), Xantphos (92 mg, 0.16 mmol) and DIEA were added (619 mg, 4.8 mmol). Under nitrogen protection, the temperature was raised to 120 °C and stirred for 1 h. After the TLC reaction was completed, the reaction mixture was added to 75 ml of saturated sodium chloride solution, then extracted with EtOAc (25 ml × 3), the organic phases were combined, and the saturated sodium chloride solution was used. The solution was washed (100ml), dried over anhydrous sodium sulfate, concentrated by spin drying, and the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05 % ammonia water), B: acetonitrile); flow rate: 25 ml/min) to give compound 8C (412 mg, pale yellow solid) in 32.6% yield.

MS(ESI):m/z 789.2[M+H] ⁺。 MS (ESI): m/z 789.2 [M+H] ⁺ .

第三步：合成化合物8Step 3: Synthesis of Compound 8

将化合物8C(354mg,0.45mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到化合物8的盐酸盐(283mg,白色固体)，产率91.5％。Compound 8C (354 mg, 0.45 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, it was filtered, and the solid was washed with ethyl acetate. Washed and dried to obtain the hydrochloride salt of compound 8 (283 mg, white solid) in 91.5% yield.

MS(ESI):m/z 689.2[M+H] ⁺。 MS(ESI): m/z 689.2 [M+H] ⁺ .

实施例9：化合物9的制备Example 9: Preparation of Compound 9

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物9AStep 1: Synthesis of Compound 9A

将化合物8C(1g,1.3mmol)加入到甲醇(5.0ml)和水(10.0ml)的混合溶剂中，然后加入一水合氢氧化锂(170mg,4.2mmol)，室温搅拌2h，TLC显示反应结束后，用1M盐酸调PH值到3-4，有大量固体析出，过滤，滤饼干燥，得到中间体9A(907mg,灰白色固体)，产率90％。粗品未经纯化，直接用于下一步反应。Compound 8C (1 g, 1.3 mmol) was added to a mixed solvent of methanol (5.0 ml) and water (10.0 ml), then lithium hydroxide monohydrate (170 mg, 4.2 mmol) was added, and the mixture was stirred at room temperature for 2 h. TLC showed that the reaction was over. , adjusted the pH to 3-4 with 1M hydrochloric acid, a large amount of solid was precipitated, filtered, and the filter cake was dried to obtain Intermediate 9A (907 mg, off-white solid) in a yield of 90%. The crude product was used directly in the next reaction without purification.

第二步：合成化合物9Step 2: Synthesis of Compound 9

将化合物9A(900mg,1.16mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，将反应混合物加入到75ml饱和碳酸氢钠溶液中，然用二氯甲烷：甲醇(5：1，V：V)萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物9(123mg,白色固体)，15.7％产率。 Compound 9A (900 mg, 1.16 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, the reaction mixture was added to 75 ml of saturated solution. sodium bicarbonate solution, then extracted with dichloromethane:methanol (5:1, V:V) (25ml×3), combined organic phases, washed with saturated sodium chloride solution (100ml), dried over anhydrous sodium sulfate, Spin dry and concentrate, and the residue is separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05% ammonia water), B: acetonitrile); flow rate: 25ml /min) to give compound 9 (123 mg, white solid) in 15.7% yield.

MS(ESI):m/z 675.1[M+H] ⁺。 MS(ESI): m/z 675.1 [M+H] ⁺ .

¹H NMR(400MHz,DMSO)δ12.30(s,1H),8.39-8.32(m,2H),8.26-8.19(m,1H),7.56(d,J＝7.2Hz,1H),7.39-7.28(m,2H),7.27-7.16(m,2H),6.86-6.73(m,2H),6.53(s,1H),4.86(d,J＝9.2Hz,1H),3.95-3.73(m,5H),3.50(s,1H),3.12(d,J＝16.8Hz,1H),2.92-2.84(m,2H),2.76-2.72(m,1H),2.04-1.85(m,3H),1.85-1.74(m,2H),1.74-1.57(m,3H)。 ¹ H NMR (400MHz, DMSO) δ 12.30(s, 1H), 8.39-8.32(m, 2H), 8.26-8.19(m, 1H), 7.56(d, J=7.2Hz, 1H), 7.39-7.28 (m,2H),7.27-7.16(m,2H),6.86-6.73(m,2H),6.53(s,1H),4.86(d,J=9.2Hz,1H),3.95-3.73(m,5H) ), 3.50(s, 1H), 3.12(d, J=16.8Hz, 1H), 2.92-2.84(m, 2H), 2.76-2.72(m, 1H), 2.04-1.85(m, 3H), 1.85- 1.74 (m, 2H), 1.74-1.57 (m, 3H).

实施例10：化合物10的制备Example 10: Preparation of Compound 10

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物10AStep 1: Synthesis of Compound 10A

将化合物9A(900mg,1.16mmol)加入到DMF(10ml)溶液中，然后加入HATU(661mg,1.74mmol)和DIEA(300mg,2.32mmol)，室温搅拌半小时后，再加入氯化铵(320mg,5.8mmol)，然后室温反应过夜，TLC显示反应结束后，将反应液加入到75ml饱和碳酸氢钠溶液中，然用二氯甲烷：甲醇(5：1，V：V)萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物10A(303mg,白色固体)，33.8％产率。 Compound 9A (900 mg, 1.16 mmol) was added to DMF (10 ml) solution, then HATU (661 mg, 1.74 mmol) and DIEA (300 mg, 2.32 mmol) were added, and after stirring at room temperature for half an hour, ammonium chloride (320 mg, 2.32 mmol) was added. 5.8mmol), and then reacted at room temperature overnight. After TLC showed that the reaction was over, the reaction solution was added to 75ml of saturated sodium bicarbonate solution, and then extracted with dichloromethane:methanol (5:1, V:V) (25ml×3) , the organic phases were combined, washed with saturated sodium chloride solution (100ml), dried over anhydrous sodium sulfate, spin-dried and concentrated, and the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36 -67% B (A: water (0.05% ammonia water), B: acetonitrile); flow rate: 25 ml/min) to give compound 10A (303 mg, white solid) in 33.8% yield.

MS(ESI):m/z 774.1[M+H] ⁺。 MS (ESI): m/z 774.1 [M+H] ⁺ .

第二步：合成化合物10Step 2: Synthesis of Compound 10

将化合物10A(300mg,0.38mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到化合物10的盐酸盐(232mg,白色固体)，产率90.5％。Compound 10A (300 mg, 0.38 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, it was filtered, and the solid was washed with ethyl acetate. After washing and drying, the hydrochloride salt of compound 10 (232 mg, white solid) was obtained in a yield of 90.5%.

MS(ESI):m/z 674.2[M+H] ⁺。 MS(ESI): m/z 674.2 [M+H] ⁺ .

¹H NMR(400MHz,MeOD)δ8.74-8.62(m,1H),8.44-8.29(m,1H),8.23-8.13(m,1H),7.78(s,1H),7.71-7.57(m,1H),7.36-7.19(m,1H),7.14-7.03(m,1H),4.64(s,1H),4.15-3.95 (m,3H),3.77-3.72(m,2H),3.66-3.59(m,3H),3.52-3.40(m,1H),3.12-3.012(m,1H),2.13-1.90(m,5H),1.81(d,J＝11.6Hz,1H),1.69(d,J＝11.65Hz,1H),1.64-1.54(m,1H)。 ¹ H NMR(400MHz,MeOD)δ8.74-8.62(m,1H),8.44-8.29(m,1H),8.23-8.13(m,1H),7.78(s,1H),7.71-7.57(m, 1H),7.36-7.19(m,1H),7.14-7.03(m,1H),4.64(s,1H),4.15-3.95(m,3H),3.77-3.72(m,2H),3.66-3.59( m,3H),3.52-3.40(m,1H),3.12-3.012(m,1H),2.13-1.90(m,5H),1.81(d,J=11.6Hz,1H),1.69(d,J= 11.65Hz, 1H), 1.64-1.54 (m, 1H).

实施例11：化合物11的制备Example 11: Preparation of Compound 11

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物11BStep 1: Synthesis of Compound 11B

将化合物Q1(1.0g,4.9mmol)加入到DMF(10ml)中，然后加入化合物11A(2.74g,9.8mmol)和BOP(4.3g,9.8mmol)，加完后室温下缓慢加入DBU(2.2g,14.7mmol)，然后室温反应过夜，反应结束后，将Boc ₂O(1.6g，7.2mmol)加入溶液中。将反应在50℃下搅拌2小时。TLC显示反应完成。在减压下浓缩混合物。残余物过硅胶层析柱(洗脱剂:石油醚/乙酸乙酯＝1/2,V/V)纯化，得到化合物11B(1.2g,淡黄色固体)，45.2％产率。 Compound Q1 (1.0 g, 4.9 mmol) was added to DMF (10 ml), then compound 11A (2.74 g, 9.8 mmol) and BOP (4.3 g, 9.8 mmol) were added, and DBU (2.2 g) was slowly added at room temperature after the addition was complete. , 14.7 mmol), and then reacted at room temperature overnight. After the reaction was completed, Boc ₂ O (1.6 g, 7.2 mmol) was added to the solution. The reaction was stirred at 50°C for 2 hours. TLC showed the reaction was complete. The mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=1/2, V/V) to obtain compound 11B (1.2 g, pale yellow solid) in 45.2% yield.

MS(ESI):m/z 546.2[M+H] ⁺。 MS (ESI): m/z 546.2 [M+H] ⁺ .

第二步：合成化合物11CStep 2: Synthesis of compound 11C

将化合物11B(872mg,1.6mmol)溶于NMP(10ml)中，加入中间体Q4(895mg,2.4mmol)、Pd ₂(dba) ₃(145mg,0.16mmol)、Xantphos(92mg,0.16mmol)和DIEA(619mg,4.8mmol)。在氮气保护下升温到120℃并搅拌1h后，TLC反应结束后，将反应混合物加入到75ml饱和氯化钠溶液中，然用EtOAc萃取(25ml×3)，合并有机相，用饱和氯化钠溶液洗涤(100ml)，无水硫酸钠干燥，旋干浓缩，残余物用制备高效液相色谱分离纯化(Welch Xtrimate C ₁₈ 150*30mm*5μM；条件：36-67％B(A:水(0.05％氨水),B:乙腈)；流速：25ml/min)，得到化合物11C(398mg,淡黄色固体)，30.5％产率。 Compound 11B (872 mg, 1.6 mmol) was dissolved in NMP (10 ml), intermediate Q4 (895 mg, 2.4 mmol), Pd2(dba ₎₃ ₍ 145 mg, 0.16 mmol), Xantphos (92 mg, 0.16 mmol) and DIEA were added (619 mg, 4.8 mmol). Under nitrogen protection, the temperature was raised to 120 °C and stirred for 1 h. After the TLC reaction was completed, the reaction mixture was added to 75 ml of saturated sodium chloride solution, then extracted with EtOAc (25 ml × 3), the organic phases were combined, and the saturated sodium chloride solution was used. The solution was washed (100ml), dried over anhydrous sodium sulfate, concentrated by spin drying, and the residue was separated and purified by preparative high performance liquid chromatography (Welch Xtrimate C ₁₈ 150*30mm*5μM; conditions: 36-67% B (A: water (0.05 % ammonia water), B: acetonitrile); flow rate: 25 ml/min) to give compound 11C (398 mg, pale yellow solid) in 30.5% yield.

MS(ESI):m/z 817.2[M+H] ⁺。 MS(ESI): m/z 817.2 [M+H] ⁺ .

第三步：合成化合物11Step 3: Synthesis of Compound 11

将化合物11C(367mg,0.45mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到化合物11的盐酸盐(295mg,白色固体)，91.5％产率。Compound 11C (367 mg, 0.45 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, it was filtered, and the solid was washed with ethyl acetate. Washing and drying gave the hydrochloride salt of compound 11 (295 mg, white solid) in 91.5% yield.

MS(ESI):m/z 717.2[M+H] ⁺。 MS (ESI): m/z 717.2 [M+H] ⁺ .

¹H NMR(400MHz,DMSO)δ12.26(s,1H),8.76(s,2H),8.60(d,J＝4.4Hz,1H),8.21(dd,J＝24.4,7.6Hz,2H),7.47(dd,J＝7.2,5.6Hz,1H),7.43-7.34(m,1H),7.30(t,J＝8.0Hz,1H),6.69(d,J＝8.0Hz,1H),4.56-4.46(m,1H),4.30(q,J＝7.2Hz,2H),3.97(d,J＝13.2Hz,1H),3.89-3.84(m,2H),3.80(s,1H),3.36-3.23(m,3H),3.15(d,J＝17.2Hz,1H),2.89(t,J＝6.4Hz,2H),2.45(s,3H),1.96-1.87(m,2H),1.87-1.76(m,3H),1.75-1.64(m,2H),1.56(d,J＝13.2Hz,1H),1.26(t,J＝7.2Hz,3H)。 ¹ H NMR (400MHz, DMSO) δ 12.26(s, 1H), 8.76(s, 2H), 8.60(d, J=4.4Hz, 1H), 8.21(dd, J=24.4, 7.6Hz, 2H), 7.47(dd,J=7.2,5.6Hz,1H),7.43-7.34(m,1H),7.30(t,J=8.0Hz,1H),6.69(d,J=8.0Hz,1H),4.56-4.46 (m,1H),4.30(q,J=7.2Hz,2H),3.97(d,J=13.2Hz,1H),3.89-3.84(m,2H),3.80(s,1H),3.36-3.23( m, 3H), 3.15(d, J=17.2Hz, 1H), 2.89(t, J=6.4Hz, 2H), 2.45(s, 3H), 1.96-1.87(m, 2H), 1.87-1.76(m , 3H), 1.75-1.64 (m, 2H), 1.56 (d, J=13.2Hz, 1H), 1.26 (t, J=7.2Hz, 3H).

实施例12：化合物12的制备Example 12: Preparation of Compound 12

具体合成路线如下：The specific synthetic route is as follows:

第一步：合成化合物12AStep 1: Synthesis of Compound 12A

第二步：合成化合物12BStep 2: Synthesis of Compound 12B

MS(ESI):m/z 774.1[M+H] ⁺。 MS (ESI): m/z 774.1 [M+H] ⁺ .

第三步：合成化合物12Step 3: Synthesis of Compound 12

将化合物11C(367mg,0.45mmol)加入到乙酸乙酯(5ml)中，再加入4M的HCl/乙酸乙酯溶液(10ml)，室温反应1h，TLC显示反应完成后，过滤，固体用乙酸乙酯洗涤，烘干，得到化合物11的盐酸盐(295mg,白色固体)，产率91.5％。Compound 11C (367 mg, 0.45 mmol) was added to ethyl acetate (5 ml), then 4M HCl/ethyl acetate solution (10 ml) was added, and the reaction was carried out at room temperature for 1 h. After TLC showed that the reaction was completed, it was filtered, and the solid was washed with ethyl acetate. Washed and dried to obtain the hydrochloride salt of compound 11 (295 mg, white solid) in 91.5% yield.

MS(ESI):m/z 717.2[M+H] ⁺。 MS (ESI): m/z 717.2 [M+H] ⁺ .

¹H NMR(400MHz,DMSO)δ12.25(s,1H),8.63-8.50(m,3H),8.21(d,J＝8.4Hz,1H),8.05(d,J＝7.6Hz,1H),7.84(s,1H),7.56(s,1H),7.40(dd,J＝7.6,5.2Hz,1H),7.29(t,J＝8.0Hz,1H),6.66(d,J＝8.0Hz,1H),4.52-4.48(m,1H),4.07(d,J＝13.2Hz,1H),3.96(d,J＝13.2 Hz,1H),3.86(t,J＝6.0Hz,2H),3.74-3.64(m,2H),3.28-3.25(m,1H),3.11(d,J＝17.2Hz,1H),2.88(t,J＝6.4Hz,2H),2.43(s,3H),2.02-1.96(m,1H),1.94-1.85(m,2H),1.85-1.75(m,3H),1.63(d,J＝11.6Hz,1H),1.53(d,J＝13.2Hz,1H). ¹ H NMR (400MHz, DMSO) δ12.25(s, 1H), 8.63-8.50(m, 3H), 8.21(d, J=8.4Hz, 1H), 8.05(d, J=7.6Hz, 1H), 7.84(s, 1H), 7.56(s, 1H), 7.40(dd, J=7.6, 5.2Hz, 1H), 7.29(t, J=8.0Hz, 1H), 6.66(d, J=8.0Hz, 1H) ),4.52-4.48(m,1H),4.07(d,J＝13.2Hz,1H),3.96(d,J＝13.2 Hz,1H),3.86(t,J＝6.0Hz,2H),3.74-3.64 (m,2H),3.28-3.25(m,1H),3.11(d,J=17.2Hz,1H),2.88(t,J=6.4Hz,2H),2.43(s,3H),2.02-1.96( m,1H),1.94-1.85(m,2H),1.85-1.75(m,3H),1.63(d,J=11.6Hz,1H),1.53(d,J=13.2Hz,1H).

实验例1：本发明化合物针对NCI-H358细胞的抗增殖活性测试Experimental Example 1: Anti-proliferative activity test of the compounds of the present invention against NCI-H358 cells

实验材料：Experimental Materials:

DMEM培养基，盘尼西林/链霉素抗生素购自维森特。胎牛血清购自Biosera。3D CellTiter-Glo(细胞活率化学发光检测试剂)试剂购自Promega。H358细胞系购自南京科佰生物科技有限公司。Envision多标记分析仪(PerkinElmer)。DMEM medium, penicillin/streptomycin antibiotics were purchased from Vicente. Fetal bovine serum was purchased from Biosera. 3D CellTiter-Glo (Cell Viability Chemiluminescence Detection Reagent) reagent was purchased from Promega. The H358 cell line was purchased from Nanjing Kebai Biotechnology Co., Ltd. Envision Multilabel Analyzer (PerkinElmer).

实验方法：experimental method:

将H358细胞种于超低吸附96孔U型板中，80μL细胞悬液每孔，其中包含3000个H358细胞。细胞板置于二氧化碳培养箱中过夜培养。H358 cells were seeded in ultra-low adsorption 96-well U-shaped plates, 80 μL of cell suspension per well, which contained 3000 H358 cells. Cell plates were incubated overnight in a carbon dioxide incubator.

将待测化合物用排枪进3倍稀释至第8个浓度，即从0.2mM稀释至91.44nM，设置双复孔实验。向中间板中加入78μL培养基，再按照对应位置，转移2μL每孔的梯度稀释化合物至中间板，混匀后转移20μL每孔到细胞板中。转移到细胞板中的化合物浓度范围是1μM至0.457nM。细胞板置于二氧化碳培养箱中培养5天。The compound to be tested was diluted 3-fold to the 8th concentration with a row gun, that is, from 0.2 mM to 91.44 nM, and a double-well experiment was set up. Add 78 μL of medium to the middle plate, and then transfer 2 μL of each well of the compound to the middle plate according to the corresponding position. After mixing, transfer 20 μL of each well to the cell plate. Compound concentrations transferred to the cell plate ranged from 1 μM to 0.457 nM. The cell plates were placed in a carbon dioxide incubator for 5 days.

向细胞板中加入每100μL的细胞活率化学发光检测试剂，室温孵育10分钟使发光信号稳定。采用多标记分析仪读数。Add every 100 μL of cell viability chemiluminescence detection reagent to the cell plate, and incubate at room temperature for 10 minutes to stabilize the luminescence signal. Read using a multi-label analyzer.

数据分析：data analysis:

利用方程式(Sample-Min)/(Max-Min)*100％将原始数据换算成抑制率，IC ₅₀的值即可通过四参数进行曲线拟合得出(GraphPad Prism中"log(inhibitor)vs.response--Variable slope"模式得出)。表1提供了本发明的化合物对H358细胞增殖的抑制活性。 Using the equation (Sample-Min)/(Max-Min)*100% to convert the raw data into inhibition rate, the IC ₅₀ value can be obtained by curve fitting with four parameters ("log(inhibitor) vs. response--Variable slope" mode). Table 1 provides the inhibitory activity of the compounds of the present invention on H358 cell proliferation.

表1.本发明中的化合物的抗细胞增殖活性数据(IC ₅₀) Table 1. Anti-cell proliferative activity data ( _IC50 ) of compounds of the present invention

化合物编号Compound number	NCI-H358抗细胞增殖活性，IC ₅₀(单位：nM) Anti-cell proliferation activity of NCI-H358, IC ₅₀ (unit: nM)
RMC-4550RMC-4550	6464
TNO-155TNO-155	6565
化合物1Compound 1	1.511.51
化合物2Compound 2	2.42.4
化合物3Compound 3	29.829.8
化合物4Compound 4	56.656.6
化合物5Compound 5	48.948.9
化合物6Compound 6	107107
化合物7Compound 7	111111
化合物8Compound 8	3.693.69
化合物9Compound 9	167167
化合物10Compound 10	2.842.84
化合物11Compound 11	128128
化合物12Compound 12	2.342.34

从表1中的实验结果可以看出，本发明中的化合物在抑制肺癌细胞株NCI-H358增殖方面具有良好的活性。部分化合物的活性超过TNO-155活性40倍。显示出极其重要的抗肿瘤潜力。It can be seen from the experimental results in Table 1 that the compounds of the present invention have good activity in inhibiting the proliferation of lung cancer cell line NCI-H358. The activity of some compounds exceeds that of TNO-155 by 40 times. Shows extremely important antitumor potential.

实验例2：本发明化合物针对MV-4-11细胞的抗增殖活性测试Experimental Example 2: Test of the antiproliferative activity of the compounds of the present invention against MV-4-11 cells

实验材料：Experimental Materials:

IMDM培养基，胎牛血清，盘尼西林/链霉素抗生素购自Promega(Madison,WI)。MV-4-11细胞系购自中国科学院细胞库。Envision多标记分析仪(PerkinElmer)。IMDM medium, fetal bovine serum, penicillin/streptomycin antibiotics were purchased from Promega (Madison, WI). The MV-4-11 cell line was purchased from the Cell Bank of the Chinese Academy of Sciences. Envision Multilabel Analyzer (PerkinElmer).

实验方法：experimental method:

将MV-4-11细胞种于白色96孔板中，80μL细胞悬液每孔，其中包含6000个MV-4-11细胞。细胞板置于二氧化碳培养箱中过夜培养。MV-4-11 cells were seeded in a white 96-well plate, 80 μL of cell suspension per well, which contained 6000 MV-4-11 cells. Cell plates were incubated overnight in a carbon dioxide incubator.

将待测化合物用排枪进行3倍稀释至第9个浓度，即从2mM稀释至304nM，设置双复孔实验。向中间板中加入78μL培养基，再按照对应位置，转移2μL每孔的梯度稀释化合物至中间板，混匀后转移20μL每孔到细胞板中。转移到细胞板中的化合物浓度范围是10μM至1.52nM。细胞板置于二氧化碳培养箱中培养3天。The compound to be tested was diluted 3-fold to the ninth concentration, that is, from 2 mM to 304 nM, and a double-well experiment was set up. Add 78 μL of medium to the middle plate, and then transfer 2 μL of each well of the compound to the middle plate according to the corresponding position. After mixing, transfer 20 μL of each well to the cell plate. Compound concentrations transferred to cell plates ranged from 10 [mu]M to 1.52 nM. The cell plates were placed in a carbon dioxide incubator for 3 days.

向细胞板中加入每孔25uL的Promega CellTiter-Glo试剂，室温孵育10分钟使发光信号稳定。采用PerkinElmer Envision多标记分析仪读数。Add 25uL of Promega CellTiter-Glo reagent per well to the cell plate and incubate at room temperature for 10 minutes to stabilize the luminescence signal. Readings were performed on a PerkinElmer Envision multi-label analyzer.

数据分析：data analysis:

利用方程式(Sample-Min)/(Max-Min)*100％将原始数据换算成抑制率，IC ₅₀的值即可通过四参数进行曲线拟合得出(GraphPad Prism中"log(inhibitor)vs.response--Variable slope"模式得出)。表2提供了本发明的化合物对MV-4-11细胞增殖的抑制活性。 Using the equation (Sample-Min)/(Max-Min)*100% to convert the raw data into inhibition rate, the IC ₅₀ value can be obtained by curve fitting with four parameters ("log(inhibitor) vs. response--Variable slope" mode). Table 2 provides the inhibitory activity of the compounds of the present invention on the proliferation of MV-4-11 cells.

表2.本发明中的化合物的抗细胞增殖活性数据(IC ₅₀) Table 2. Anti-cell proliferative activity data ( _IC50 ) of compounds of the present invention

从表2中的实验结果可以看出，本发明中的化合物在抑制MV-4-11细胞增殖方面具有良好的活性。化合物1的活性超过TNO-155活性10倍。显示出极其重要的抗肿瘤潜力。It can be seen from the experimental results in Table 2 that the compounds of the present invention have good activity in inhibiting the proliferation of MV-4-11 cells. The activity of compound 1 exceeded that of TNO-155 by a factor of 10. Shows extremely important antitumor potential.

实验例3：p-ERK活性抑制实验Experimental example 3: p-ERK activity inhibition experiment

1.实验材料1. Experimental materials

H358细胞购自南京科佰生物科技；H358 cells were purchased from Nanjing Kebai Biotechnology;

1640培养基购自Biological industries；1640 medium was purchased from Biological industries;

胎牛血清购自Biosera；Fetal bovine serum was purchased from Biosera;

磷酸化pERK(磷酸化位点202/204)检测试剂盒购自Cisbio。Phosphorylated pERK (phosphorylation sites 202/204) detection kit was purchased from Cisbio.

2.实验方法2. Experimental method

将H358细胞种于透明96孔细胞培养板中，80μL细胞悬液/孔，每孔包含10000个H358细胞。将细胞板放入二氧化碳培养箱中，于37℃过夜孵育。弃掉细胞上清液，加入80μL/孔的饥饿培养基(1640+0.02％胎牛血清+1％双抗)，将细胞板放入二氧化碳培养箱中，细胞饥饿处理过夜。H358 cells were seeded in a transparent 96-well cell culture plate, 80 μL of cell suspension/well, and each well contained 10,000 H358 cells. Place the cell plate in a carbon dioxide incubator and incubate overnight at 37°C. The cell supernatant was discarded, 80 μL/well of starvation medium (1640+0.02% fetal bovine serum+1% double antibody) was added, the cell plate was placed in a carbon dioxide incubator, and the cells were starved overnight.

将待测化合物用100％DMSO稀释到4mM，作为第一个浓度，然后再用排枪移液器进行5倍稀释至第8个浓度，即从4mM稀释至10.24μM。取1μL化合物加入79μL细胞饥饿培养基，混匀后转移20μL/孔的化合物溶液到对应细胞板孔中，细胞板放回二氧化碳培养箱继续孵育1h，此时化合物浓度为10μM至0.0256nM，DMSO浓度为0.25％；The compounds to be tested were diluted with 100% DMSO to 4 mM as the first concentration, and then 5-fold diluted to the eighth concentration, ie, from 4 mM to 10.24 [mu]M, using a guillotine pipette. Add 1 μL of compound to 79 μL of cell starvation medium, mix well and transfer 20 μL/well of compound solution to the corresponding cell plate wells, put the cell plate back into the carbon dioxide incubator and continue to incubate for 1 h. At this time, the compound concentration is 10 μM to 0.0256nM, and the DMSO concentration is 0.25%;

结束孵育后，弃掉细胞上清液，加入50μL细胞裂解液/孔，室温摇晃孵育30min；使用检测缓冲液将铕穴状化合物标记的磷酸化细胞外调节蛋白激酶抗体和d2标记的磷酸化细胞外调节蛋白激酶抗体稀释20倍；取16μL细胞裂解物上清液，加入到新的384白色微孔板中，再加入2μL铕穴状化合物标记的磷酸化细胞外调节蛋白激酶抗体稀释液和2μL d2标记的磷酸化细胞外调节蛋白激酶抗体稀释液，常温孵育4h；孵育结束后，使用多标记分析仪读取均相时间分辨荧光HTRF(激发波长为320nm，发射波长为615nm和665nm)。After the incubation, discard the cell supernatant, add 50 μL of cell lysate/well, and incubate with shaking at room temperature for 30 min; use detection buffer to detect the phosphorylated extracellular regulatory protein kinase antibody labeled with europium cryptate and the phosphorylated cells labeled with d2. 20-fold dilution of the extracellular regulatory protein kinase antibody; take 16 μL of cell lysate supernatant and add it to a new 384 white microplate, then add 2 μL of europium cryptate-labeled phosphorylated extracellular regulatory protein kinase antibody dilution and 2 μL D2-labeled phosphorylated extracellular regulated protein kinase antibody dilution was incubated at room temperature for 4 h; after incubation, homogeneous time-resolved fluorescence HTRF was read using a multi-label analyzer (excitation wavelength 320 nm, emission wavelengths 615 nm and 665 nm).

3.数据分析3. Data Analysis

利用方程式(样品-Min)/(Max-Min)成100％，将原始数据换算成抑制率，IC ₅₀值即可通过四参数进行曲线拟合而得出(例如，通过GraphPad Prism中的“log(inhibitor)vs.response--Variable slope”模式得出)。其中，Max孔：阳性对照孔读值为1X细胞裂解液；Min孔：阴性对照孔读值为0.25％DMSO细胞孔细胞裂解液。 Using the equation (Sample-Min)/(Max-Min) to 100%, converting raw data to inhibition, _IC50 values can be obtained by curve fitting with four parameters (eg, by "log" in GraphPad Prism (inhibitor)vs.response--Variable slope" mode). Among them, Max well: the reading value of the positive control well is 1X cell lysate; Min well: the reading value of the negative control well is 0.25% DMSO cell well cell lysate.

表3.本发明中的化合物的p-ERK活性抑制数据(IC ₅₀) Table 3. p-ERK activity inhibition data ( _IC50 ) for compounds of the present invention

化合物编号Compound number	p-ERK抑制活性，IC ₅₀(单位：nM) p-ERK inhibitory activity, IC ₅₀ (unit: nM)
TNO-155TNO-155	85.7685.76
化合物1Compound 1	9.539.53

从表3中的实验结果可以看出，本发明中的化合物对NCI-H358细胞中SHP2下游Erk磷酸化具有优异的抑制效果，进一步从机制上阐述了本发明中的化合物具备较好的抗肿瘤前景。From the experimental results in Table 3, it can be seen that the compounds of the present invention have excellent inhibitory effect on the phosphorylation of Erk downstream of SHP2 in NCI-H358 cells, and it is further explained from the mechanism that the compounds of the present invention have better anti-tumor effects prospect.

实验例4：SHP2酶学实验Experimental Example 4: SHP2 Enzymatic Experiment

1.实验材料和仪器1. Experimental Materials and Instruments

均相全长SHP2酶学实验试剂盒购自BPS Bioscience；Homogeneous full-length SHP2 enzymatic assay kit was purchased from BPS Bioscience;

多标记分析仪购自Perkin Elmer。A multi-label analyzer was purchased from Perkin Elmer.

2.实验方法：2. Experimental method:

将5X检测缓冲液用去离子水稀释成1X检测缓冲液，现配现用，配好后放置在冰上，备用。Dilute the 5X detection buffer with deionized water to 1X detection buffer, prepare it for immediate use, and place it on ice for later use.

将待测化合物用100％DMSO稀释到100μM，作为第一个浓度，然后再用排枪移液器进行4倍稀释至第8个浓度，即从100μM稀释至6.1nM。用1X缓冲液将待测化合物各梯度稀释成含10％DMSO的工作液，5μL/孔加到对应孔中，设置双复孔实验。1000rpm离心1min。Compounds to be tested were diluted with 100% DMSO to 100 μM as the first concentration, and then 4-fold diluted to the 8th concentration with a guillotine pipette, ie, from 100 μM to 6.1 nM. Each compound to be tested was diluted with 1X buffer into a working solution containing 10% DMSO, and 5 μL/well was added to the corresponding well to set up a double-well experiment. Centrifuge at 1000 rpm for 1 min.

每孔加入18μL配制的反应混合液，其中含12.25μL去离子水；5μL 5X检测缓冲液；0.25μL蛋白酪氨酸磷酸酶激活多肽(100μM)；0.5μL DTT(250mM)。1000rpm离心1min。Add 18 μL of the prepared reaction mixture to each well, which contains 12.25 μL deionized water; 5 μL 5X detection buffer; 0.25 μL protein tyrosine phosphatase activating polypeptide (100 μM); 0.5 μL DTT (250 mM). Centrifuge at 1000 rpm for 1 min.

用1X检测缓冲液将SHP2酶稀释到0.1ng/μL，取2μL/孔加入到对应孔中，阴性对照孔中加入2μL 1X检测缓冲液，SHP2(0.2ng)，该步在冰上操作，反应体系置于25℃孵育60min，做化合物预孵育。Dilute the SHP2 enzyme to 0.1ng/μL with 1X detection buffer, add 2μL/well to the corresponding well, add 2μL 1X detection buffer, SHP2 (0.2ng) to the negative control well, this step is operated on ice, the reaction The system was incubated at 25°C for 60 min as compound pre-incubation.

化合物预孵育结束后，每孔加入25μL底物工作液，其中含19.45μL去离子水；5μL5X检测缓冲液；0.5μL DTT(250mM)和0.05μL SHP2底物(DiFMUP)(10mM)，反应体系置于25℃反应30min。此时化合物终浓度梯度为1μM至0.061nM。反应结束后，采用多标记分析仪读取荧光值(激发波长为360nm，发射波长为460nm)。After compound pre-incubation, 25 μL of substrate working solution was added to each well, which contained 19.45 μL of deionized water; 5 μL of 5X detection buffer; 0.5 μL of DTT (250 mM) and 0.05 μL of SHP2 substrate (DiFMUP) (10 mM). React at 25°C for 30min. The final compound concentration gradient at this point was 1 μM to 0.061 nM. After the reaction, the fluorescence value was read by a multi-label analyzer (excitation wavelength was 360 nm, emission wavelength was 460 nm).

化合物本底读值的检测方法如下：取100％DMSO稀释好的待测化合物各梯度5μL到新的化合物板中，加入45μL 1X检测缓冲液进行10倍稀释，配成10％DMSO的工作液，再取该化合物工作液5μL/孔到检测板中，然后加入45μL 1X检测缓冲液进行10倍稀释，此时DMSO终浓度为1％，1000rpm离心1分钟，采用多标记分析仪读取荧光值(激发波长为360nm，发射波长为460nm)。The detection method of compound background reading value is as follows: take 5 μL of each compound to be tested diluted in 100% DMSO into a new compound plate, add 45 μL of 1X detection buffer for 10-fold dilution, and prepare a working solution of 10% DMSO, Then take 5 μL/well of the compound working solution into the detection plate, then add 45 μL 1X detection buffer for 10-fold dilution, at this time the final concentration of DMSO is 1%, centrifuge at 1000 rpm for 1 minute, and use a multi-label analyzer to read the fluorescence value ( Excitation wavelength is 360 nm, emission wavelength is 460 nm).

3.数据分析3. Data Analysis

以酶反应原始读值减去化合物本底读值后得到的数据作为抑制率计算的初始值，利用方程式(样品-Min)/(Max-Min)×100％，将初始值换算成抑制率，IC ₅₀值即可通过四参数进行曲线拟合而得出(例如，通过GraphPad Prism中的“log(inhibitor)vs.response--Variable slope”模式得出)。其中，Max孔：阳性对照孔初始值；Min孔：阴性对照孔初始值。 The data obtained by subtracting the background reading of the compound from the original reading of the enzyme reaction was used as the initial value for the calculation of the inhibition rate, and the initial value was converted into the inhibition rate using the equation (sample-Min)/(Max-Min)×100%, _IC50 values can then be obtained by curve fitting with four parameters (eg, by the "log(inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Among them, Max well: initial value of positive control well; Min well: initial value of negative control well.

表4.本发明中的化合物的SHP2酶活性抑制数据(IC ₅₀) Table 4. SHP2 enzymatic activity inhibition data ( _IC50 ) for compounds of the present invention

化合物编号Compound number	SHP2酶抑制活性IC ₅₀(单位：nM) SHP2 enzyme inhibitory activity IC ₅₀ (unit: nM)
TNO-155TNO-155	2.62.6
化合物1Compound 1	0.980.98

从表4中的实验结果可以看出，本发明中的化合物在酶学层面上显示出非常优秀的SHP2酶抑制活性，IC ₅₀达到1nM以下，极具开发及应用前景。 It can be seen from the experimental results in Table 4 that the compounds in the present invention show very good SHP2 enzyme inhibitory activity at the enzymatic level, with IC ₅₀ reaching below 1 nM, which is very promising for development and application.

实验例5：hERG钾离子通道抑制剂活性测试Experimental example 5: hERG potassium channel inhibitor activity test

1细胞准备1 cell preparation

CHO-hERG细胞培养于175cm ²培养瓶中，待细胞密度生长到60-80％，移走培养液，用7mL PBS洗一遍，然后加入3mL Detachin消化。 CHO-hERG cells were cultured in a 175cm ² culture flask. When the cell density reached 60-80%, the culture medium was removed, washed with 7mL of PBS, and then digested with 3mL of Detachin.

待消化完全后加入7mL培养液中和，然后离心，吸走上清液，加再加入5mL培养液重悬，以确保细胞密度为2-5×10 ⁶/mL。 After the digestion is complete, add 7 mL of culture medium to neutralize, then centrifuge, remove the supernatant, and add 5 mL of culture medium to resuspend to ensure that the cell density is 2-5×10 ⁶ /mL.

2溶液配制见表5：2 The solution preparation is shown in Table 5:

表5.细胞内液和外液的组成成分Table 5. Composition of intracellular and extracellular fluids

3电生理记录过程3 Electrophysiological recording process

单细胞高阴抗封接和全细胞模式形成过程全部由Qpatch仪器自动完成，在获得全细胞记录模式后，细胞钳制备在-80毫伏，在给予一个5秒的+40毫伏去极化刺激前，先给予一个50毫秒的-50毫伏前置电压，然后复极化到-50毫伏维持5秒，再回到-80毫伏。每15秒施加此电压刺激，记录2分钟后给予细胞外液记录5分钟，然后开始给药过程，化合物浓度从最低测试浓度开始，每个测试浓度给予2.5分钟，连续给完所有浓度后，给予阳性对照化合物0.1μM Cisapride。每个浓度至少测试3个细胞(n≥3)。The single-cell high-negative anti-seal and whole-cell pattern formation process are all completed automatically by the Qpatch instrument. After obtaining the whole-cell recording pattern, the cell clamp is prepared at -80 mV, and a 5-second depolarization of +40 mV is given. Before stimulation, a pre-voltage of -50 mV was given for 50 ms, followed by repolarization to -50 mV for 5 s and back to -80 mV. This voltage stimulus was applied every 15 seconds, and the extracellular fluid was recorded for 2 minutes and then recorded for 5 minutes. Then the dosing process was started. The compound concentration started from the lowest test concentration, and each test concentration was administered for 2.5 minutes. After all concentrations were continuously administered, the administration Positive control compound 0.1 μM Cisapride. At least 3 cells were tested at each concentration (n≥3).

4化合物准备4 Compound Preparation

将20mM的化合物母液用细胞外液进行稀释，取5μL 20mM的化合物母液加入2495μL细胞外液，500倍稀释至40μM，然后在含0.2％DMSO的细胞外液中依次进行3倍连续稀释得到需要测试的最终浓度。Dilute the 20 mM compound stock solution with extracellular fluid, add 5 μL of 20 mM compound stock solution to 2495 μL of extracellular fluid, dilute 500 times to 40 μM, and then perform 3-fold serial dilutions in the extracellular fluid containing 0.2% DMSO to obtain the required test. the final concentration.

最高测试浓度为40μM，依次分别为40，13.33，4.44，1.48，0.49，0.16μM共6个浓度。The highest tested concentration was 40 μM, followed by 40, 13.33, 4.44, 1.48, 0.49, and 0.16 μM in a total of 6 concentrations.

最终测试浓度中的DMSO含量不超过0.2％，此浓度的DMSO对hERG钾通道没有影响。The DMSO content in the final test concentration did not exceed 0.2%, and this concentration of DMSO had no effect on the hERG potassium channel.

5数据分析5 Data Analysis

实验数据由XLFit软件进行分析。The experimental data were analyzed by XLFit software.

6质量控制6Quality Control

环境：湿度20～50％，温度22～25℃Environment: Humidity 20～50%, Temperature 22～25℃

试剂：所用实验试剂购买于Sigma公司，纯度>98％Reagents: The experimental reagents used were purchased from Sigma, the purity was >98%

报告中的实验数据必须满足以下标准：Experimental data in the report must meet the following criteria:

全细胞封接阻抗>100MΩWhole cell sealing impedance >100MΩ

尾电流幅度>300pATail current amplitude>300pA

药理学参数：Pharmacological parameters:

多浓度Cisapride对hERG通道的抑制效应设为阳性对照。The inhibitory effect of multiple concentrations of Cisapride on hERG channel was set as a positive control.

7实验结果7 Experimental results

表6.本发明中的化合物在多浓度对hERG电流的抑制结果Table 6. Inhibitory results of the compounds of the present invention on hERG current at multiple concentrations

化合物编号Compound number	hERG(μM)hERG(μM)
化合物1Compound 1	>40>40
化合物2Compound 2	>40>40
CisaprideCisapride	0.0210.021

8实验结论∶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。从实验结果可以看出，本发明实施例化合物对于心脏hERG钾离子通道没有明显抑制作用，心脏毒副作用风险低。8. Conclusions: The inhibition of cardiac hERG potassium ion channels by drugs is the main reason for drugs to cause QT prolongation syndrome. It can be seen from the experimental results that the compounds of the examples of the present invention have no obvious inhibitory effect on the cardiac hERG potassium ion channel, and the risk of cardiotoxicity and side effects is low.

实验例6：药物代谢动力学实验Experimental Example 6: Pharmacokinetic Experiment

1.实验材料1. Experimental materials

使用以上实施例中制备的化合物，口服药物配制成0.3mg/mL澄清溶液(2％DMSO+30％PEG300+2％Tween80+66％H ₂O)，静脉药物配制成0.2mg/mL澄清溶液(2％DMSO+30％PEG300+2％Tween80+66％H ₂O)。 Using the compounds prepared in the above examples, the oral drug was formulated into a 0.3 mg/mL clear solution (2% DMSO+30% PEG300+2% Tween80+66% H ₂ O), and the intravenous drug was formulated into a 0.2 mg/mL clear solution ( 2% DMSO + 30% PEG300 + 2% Tween 80 + 66% _H2O ).

2.实验动物2. Experimental animals

雄性CD-1小鼠或大鼠，每组各3只，体重27-28g，由上海斯莱克实验动物责任有限公司提供。受试小鼠实验前给予2-4天的环境适应期，给药前禁食8-12h，给药2h后给水，4h后给食。Male CD-1 mice or rats, three in each group, weighing 27-28 g, were provided by Shanghai Slack Laboratory Animal Co., Ltd. The tested mice were given a 2-4 day environmental adaptation period before the experiment, fasted for 8-12 hours before administration, given water 2 hours after administration, and given food after 4 hours.

3.实验方法3. Experimental method

1)小鼠或大鼠禁食但可自由饮水12h后，采取0时刻空白血浆；1) After the mice or rats were fasted but had free access to water for 12 hours, blank plasma was taken at time 0;

2)取步骤1)中的小鼠，口服(PO)给予待测化合物3mg/kg；静脉(IV)给予待测化合物1mg/kg；2) Take the mice in step 1), and administer 3 mg/kg of the compound to be tested orally (PO); 1 mg/kg of the compound to be tested is administered intravenously (IV);

3)于口服后5min、15min、30min、1h、2h、4h、8h、10h和24h，从眼底静脉丛连续取血，置于分布有肝素的EP管中，8000rpm离心5min后，取上层血浆，-20℃冻存，待LC-MS/MS分析；3) At 5min, 15min, 30min, 1h, 2h, 4h, 8h, 10h and 24h after oral administration, blood was continuously collected from the fundus venous plexus, placed in an EP tube distributed with heparin, centrifuged at 8000 rpm for 5min, and the upper plasma was collected. Freeze at -20°C until LC-MS/MS analysis;

4)根据步骤3)所得的血药浓度-时间数据，采用WinNonlin软件求算药代动力学参数，具体数据见表7。4) According to the blood drug concentration-time data obtained in step 3), use WinNonlin software to calculate the pharmacokinetic parameters, and the specific data are shown in Table 7.

表7.本发明中的化合物的药代动力学数据Table 7. Pharmacokinetic data for compounds of the present invention

如表7中所示，口服或静脉给予小鼠或大鼠本发明中的化合物后，在动物血浆中有较高的暴露量，清除率低，可以通过口服给药。As shown in Table 7, after oral or intravenous administration of the compounds of the present invention to mice or rats, there are higher exposures in animal plasma, low clearance, and can be administered orally.

实验例7：体内药效学实验一Experimental example 7: In vivo pharmacodynamic experiment 1

1.实验目的1. Experimental purpose

评价受试化合物在人非小细胞肺癌NCI-H358皮下异体移植肿瘤模型上的体内药效。The in vivo efficacy of the test compounds on the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model was evaluated.

2.实验动物2. Experimental animals

BALB/裸小鼠，雌性，6-8周龄，体重18-20克，共需18只，由上海灵畅实验动物有限公司提供。BALB/nude mice, female, 6-8 weeks old, weighing 18-20 grams, a total of 18 mice were required, provided by Shanghai Lingchang Laboratory Animal Co., Ltd.

3.实验方法3. Experimental method

将NCI-H358肿瘤细胞重悬于PBS中，制备成密度为5×10 ⁷个/mL的细胞悬液，皮下接种于每只小鼠的右后背(0.1mL，5×10 ⁶/只)，等待肿瘤生长。在肿瘤平均体积达到约151mm ³时，开始进行随机分组给药。给药后，每周两次用游标卡尺测量肿瘤直径，肿瘤体积的计算公式如下： The NCI-H358 tumor cells were resuspended in PBS to prepare a cell suspension with a density of 5×10 ⁷ cells/mL, and subcutaneously inoculated into the right back of each mouse (0.1 mL, 5×10 ⁶ /mice) , waiting for the tumor to grow. Randomization was initiated when the mean tumor volume reached approximately 151 ^mm3 . After administration, tumor diameters were measured with a vernier caliper twice a week, and the tumor volume was calculated as follows:

V＝0.5a×b2，其中a和b分别表示肿瘤的长径和短径。V=0.5a×b2, where a and b represent the long and short diameters of the tumor, respectively.

化合物的抑瘤疗效用TGI(％)评价，TGI(％)反映了肿瘤生长抑制率，其计算如下：The tumor-inhibitory efficacy of a compound was evaluated by TGI (%), which reflects the tumor growth inhibition rate and was calculated as follows:

TGI(％)＝[(1-(某处理组给药结束时平均瘤体积-该处理组开始给药时平均瘤体积)/(溶剂对照组治疗结束时平均瘤体积-溶剂对照组开始治疗时平均瘤体积)]×100％。TGI(%)=[(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group)/(average tumor volume at the end of treatment in the solvent control group-at the beginning of treatment in the solvent control group Mean tumor volume)] × 100%.

4.实验结果4. Experimental results

相关结果如表8所示。The related results are shown in Table 8.

表8.体内药效实验结果Table 8. In vivo efficacy test results

组别group	肿瘤体积(mm ³)(第35天) Tumor volume (mm ³ ) (day 35)	TGI(％)TGI(%)
溶剂对照组solvent control	784.14±40.83784.14±40.83	//
TNO-155(30mg/kg)TNO-155(30mg/kg)	355.37±68.17355.37±68.17	67.7267.72
化合物1(30mg/kg)Compound 1 (30mg/kg)	185.34±20.85185.34±20.85	94.5794.57

5.实验结论5. Experimental conclusion

开始给药35天后，在同等剂量(30mg/kg)下，本发明中的化合物与阳性对照物TNO-155相比，具有更显著的肿瘤抑制效果，TGI(％)达到94.57％，明显优于阳性对照物TNO155的67.72％，表明本发明中的化合物在人非小细胞肺癌NCI-H358皮下异体移植肿瘤模型上展示出良好的体内药效。作为SHP2抑制剂，在临床上有着非常好的抗肿瘤应用前景。35 days after the start of administration, under the same dose (30mg/kg), the compound of the present invention has a more significant tumor inhibitory effect than the positive control TNO-155, and the TGI (%) reaches 94.57%, which is significantly better than 67.72% of the positive control TNO155, indicating that the compounds of the present invention show good in vivo efficacy in the human non-small cell lung cancer NCI-H358 subcutaneous xenograft tumor model. As a SHP2 inhibitor, it has a very promising anti-tumor application prospect in clinical.

实验例8：体内药效学实验二Experimental example 8: In vivo pharmacodynamic experiment 2

1.实验目的1. Experimental purpose

评价受试化合物在人胰腺癌MIA-PaCa2细胞皮下异体移植肿瘤模型上的体内药效。To evaluate the in vivo efficacy of test compounds in human pancreatic cancer MIA-PaCa2 cell subcutaneous xenograft tumor model.

2.实验动物2. Experimental animals

BALB/裸小鼠，雌性，6-8周龄，体重18-22克，共需24只，由由北京维通利华科技股份有限公司提供。BALB/nude mice, female, 6-8 weeks old, weighing 18-22 grams, a total of 24 mice were required, provided by Beijing Weitong Lihua Technology Co., Ltd.

3.实验方法3. Experimental method

将MIA-PaCa2肿瘤细胞重悬于PBS中，制备成密度为1×10 ⁷个/mL的细胞悬液，皮下接种0.2mL细胞悬液于每只小鼠的右后背(加入基质胶，体积比为1：1)，等待肿瘤生长。在肿瘤平均体积达到约142mm ³时，开始进行随机分组给药。给药后，每周两次用游标卡尺测量肿瘤直径，肿瘤体积的计算公式如下： MIA-PaCa2 tumor cells were resuspended in PBS to prepare a cell suspension with a density of 1×10 ⁷ cells/mL, and 0.2 mL of the cell suspension was subcutaneously inoculated on the right back of each mouse (Add Matrigel, volume ratio of 1:1), waiting for tumor growth. Randomization was initiated when the mean tumor volume reached approximately 142 ^mm3 . After administration, tumor diameters were measured with a vernier caliper twice a week, and the tumor volume was calculated as follows:

4.实验结果4. Experimental results

相关结果如表9所示。The related results are shown in Table 9.

表9.体内药效实验结果Table 9. In vivo efficacy test results

组别group	肿瘤体积(mm ³)(第22天) Tumor volume (mm ³ ) (day 22)	TGI(％)TGI(%)
溶剂对照组solvent control	1560±1371560±137	//
TNO-155(30mg/kg)TNO-155(30mg/kg)	868±72868±72	48.848.8
化合物1(3.0mg/kg)Compound 1 (3.0mg/kg)	696±112696±112	60.960.9
化合物1(30mg/kg)Compound 1 (30mg/kg)	560±93560±93	70.470.4

5.实验结论5. Experimental conclusion

开始给药22天后，在同等剂量(30mg/kg)下，本发明中的化合物与阳性对照物TNO-155相比，具有更加显著的肿瘤抑制效果，并且有明显的量效关系，表明本发明中的化合物在人胰腺癌MIA-PaCa2皮下异体移植肿瘤模型上展示出良好的体内药效。22 days after the start of administration, under the same dose (30mg/kg), the compound of the present invention has a more significant tumor inhibitory effect compared with the positive control TNO-155, and has an obvious dose-effect relationship, indicating that the present invention The compounds in human pancreatic cancer MIA-PaCa2 subcutaneous xenograft tumor model showed good in vivo efficacy.

实验例9：体内药效学实验三Experimental Example 9: In vivo pharmacodynamic experiment 3

1.实验目的1. Experimental purpose

评价受试化合物在鼠源结肠癌MC38皮下异种移植肿瘤模型上的体内药效。The in vivo efficacy of the test compounds on the murine colon cancer MC38 subcutaneous xenograft tumor model was evaluated.

2.实验动物2. Experimental animals

C57BL/6小鼠，雌性，6～8周龄，体重18～20克。共需32只，由上海灵畅实验动物有限公司提供。C57BL/6 mice, female, 6-8 weeks old, weighing 18-20 grams. A total of 32 animals were required, provided by Shanghai Lingchang Laboratory Animal Co., Ltd.

3.实验方法3. Experimental method

将MC38肿瘤细胞重悬于PBS中，制备成密度为0.3×10 ⁶个/mL的细胞悬液，皮下接种0.1mL细胞悬液于每只小鼠的右后背(加入基质胶，体积比为1：1)，等待肿瘤生长。在肿瘤平均体积达到约65mm ³时，开始进行随机分组给药。给药后，每周两次用游标卡尺测量肿瘤直径，肿瘤体积的计算公式如下： The MC38 tumor cells were resuspended in PBS to prepare a cell suspension with a density of 0.3×10 ⁶ cells/mL, and 0.1 mL of the cell suspension was subcutaneously inoculated on the right back of each mouse (by adding Matrigel, the volume ratio was 1:1), waiting for tumor growth. Randomization was initiated when the mean tumor volume reached approximately 65 ^mm3 . After administration, tumor diameters were measured with a vernier caliper twice a week, and the tumor volume was calculated as follows:

V＝0.5a×b ²，其中a和b分别表示肿瘤的长径和短径。 V=0.5a×b ² , where a and b represent the long and short diameters of the tumor, respectively.

化合物的抑瘤疗效用TGI(％)评价，TGI(％)反映了肿瘤生长抑制率，其计算如下：The tumor-inhibitory efficacy of the compound was evaluated by TGI (%), which reflects the tumor growth inhibition rate and was calculated as follows:

4.实验结果4. Experimental results

相关结果如表10所示。The related results are shown in Table 10.

表10.体内药效实验结果Table 10. In vivo efficacy test results

5.实验结论5. Experimental conclusion

开始给药18天后，本发明中的化合物与PD-1抗体联合用药，与单药相比，明显能增加肿瘤抑制效果，在鼠源结肠癌MC38皮下异种移植肿瘤模型上的展示出良好的体内药效。表明本发明中的化合物与PD1单抗联用显示协同增效抗肿瘤作用。18 days after the start of administration, the compound of the present invention combined with PD-1 antibody can significantly increase the tumor inhibitory effect compared with the single drug, and shows good in vivo effect on the murine colon cancer MC38 subcutaneous xenograft tumor model. Efficacy. It shows that the compound of the present invention and PD1 monoclonal antibody show synergistic anti-tumor effect.

实验例10：体内药效学实验四Experimental Example 10: In Vivo Pharmacodynamic Experiment 4

1.实验目的1. Experimental purpose

评价受试化合物在人食管癌KYSE-520皮下异体移植肿瘤模型上的体内药效。The in vivo efficacy of the test compounds on the human esophageal cancer KYSE-520 subcutaneous xenograft tumor model was evaluated.

2.实验动物2. Experimental animals

BALB/裸小鼠，雌性，6-8周龄，体重18-24克，共需40只，由北京维通利华科技股份有限公司提供。BALB/nude mice, female, 6-8 weeks old, weighing 18-24 grams, a total of 40 mice were required, provided by Beijing Weitong Lihua Technology Co., Ltd.

3.实验方法3. Experimental method

将KYSE-520肿瘤细胞重悬于PBS中，制备成密度为10×10 ⁶个/mL的细胞悬液，皮下接种0.2mL细胞悬液于每只小鼠的右后背(加基质胶，体积比1：1)，等待肿瘤生长。在肿瘤平均体积达到约141mm ³时，开始进行随机分组给药。给药后，每周两次用游标卡尺测量肿瘤直径，肿瘤体积的计算公式如下： The KYSE-520 tumor cells were resuspended in PBS to prepare a cell suspension with a density of 10×10 ⁶ cells/mL, and 0.2 mL of the cell suspension was subcutaneously inoculated on the right back of each mouse (plus Matrigel, volume ratio 1:1), waiting for the tumor to grow. Randomization was initiated when the mean tumor volume reached approximately 141 ^mm3 . After administration, tumor diameters were measured with a vernier caliper twice a week, and the tumor volume was calculated as follows:

4.实验结果4. Experimental results

相关结果如表11所示。The relevant results are shown in Table 11.

表11.体内药效实验结果Table 11. In vivo efficacy test results

组别group	肿瘤体积(mm ³)(第21天) Tumor volume (mm ³ ) (day 21)	TGI(％)TGI(%)
溶剂对照组solvent control	1,059±1761,059±176	//
TNO-155(30mg/kg)TNO-155(30mg/kg)	304±36304±36	82.382.3
化合物1(0.3mg/kg)Compound 1 (0.3mg/kg)	439±69439±69	67.567.5
化合物1(1.0mg/kg)Compound 1 (1.0mg/kg)	321±45321±45	80.480.4
化合物1(3.0mg/kg)Compound 1 (3.0mg/kg)	216±42216±42	91.991.9

5.实验结论5. Experimental conclusion

开始给药21天后，化合物1在阳性对照十分之一的剂量(3.0mg/kg)条件下，本发明中的化合物1与阳性对照物TNO-155(30mg/kg)相比，具有显著的更好的肿瘤抑制效果，TGI(％)达到91.9％，明显优于阳性对照物TNO155的82.3％。另外，本发明的化合物1在更低的剂量(0.3mg/kg)时，仍展示出显著的抗肿瘤药效，这表明本发明中的化合物在人食管癌KYSE-520皮下异体移植模型中展示良好的体内药效，且抗肿瘤作用具有剂量依赖性的趋势。21 days after the start of administration, under the condition of one tenth of the dose of the positive control (3.0 mg/kg), the compound 1 of the present invention has a significant effect compared with the positive control TNO-155 (30 mg/kg). Better tumor inhibition effect, TGI (%) reached 91.9%, which was obviously better than 82.3% of the positive control TNO155. In addition, the compound 1 of the present invention still exhibits significant antitumor efficacy at a lower dose (0.3 mg/kg), which indicates that the compound of the present invention exhibits in the human esophageal cancer KYSE-520 subcutaneous xenograft model Good in vivo efficacy, and the anti-tumor effect has a dose-dependent trend.

尽管上面已经示出和描述了本发明的实施例，可以理解的是，上述实施例是示例性的，不能理解为对本发明的限制。在不脱离本发明的原理和宗旨的情况下，本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型，这些变化、修改、替换和变型均涵盖在本发明的范围之中。Although the embodiments of the present invention have been shown and described above, it is to be understood that the above-described embodiments are exemplary and should not be construed as limiting the present invention. Without departing from the principles and spirit of the present invention, those of ordinary skill in the art can make changes, modifications, substitutions and alterations to the above-mentioned embodiments within the scope of the present invention, and these changes, modifications, substitutions and modifications are all included in the within the scope of the present invention.

Claims

如式I所示的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药，其中A compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof, wherein

X ¹、X ²和X ³各自独立地选自CR ₅和N，或不存在； X ¹ , X ² and X ³ are each independently selected from CR ₅ and N, or are absent;

当X ¹、X ²和X ³各自独立地选自CR ₅和N时，X ⁵和X ⁶各自独立地选自C和N，X ⁷为CR ₅或N； When X ¹ , X ² and X ³ are each independently selected from CR ₅ and N, X ⁵ and X ⁶ are each independently selected from C and N, and X ⁷ is CR ₅ or N;

当X ¹、X ²和X ³不存在时，X ⁵、X ⁶和X ⁷各自独立地选自CR ₅和N； When X ¹ , X ² and X ³ are absent, X ⁵ , X ⁶ and X ⁷ are each independently selected from CR ₅ and N;

X ⁴为C或N； X ⁴ is C or N;

X ⁸为N或NR ₅； X ⁸ is N or NR ₅ ;

R ₁、R ₂、R ₃、R ₄、R ₆和R ₇各自独立地选自氢、卤素、羟基、氨基、氧代基、氰基、C ₂-C ₈烯基、C ₂-C ₈炔基、醛基、氨基甲酰基、C ₁-C ₈烷基、C ₁-C ₈杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₈烷氧基和C ₁-C ₃卤代烷氧基，其中所述C ₁-C ₈烷基、C ₁-C ₈杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₈烷氧基和C ₁-C ₃卤代烷氧基各自任选地被一个或多个R ₅取代； R ₁ , R ₂ , R ₃ , R ₄ , R ₆ and R ₇ are each independently selected from hydrogen, halogen, hydroxy, amino, oxo, cyano, C ₂ -C ₈ alkenyl, C ₂ -C ₈ Alkynyl, aldehyde, carbamoyl, C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₈ alkoxy and C ₁ -C ₃ haloalkoxy, wherein said C ₁ -C ₈ alkyl, C ₁ -C ₈ heteroalkyl, C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycle alkyl, _C1 - _C8alkoxy and _C1 _- C3 haloalkoxy are each optionally substituted with one or more R5 _;

若存在，每一个R ₅各自独立地选自氢、卤素、羟基、氨基、氰基、氨基甲酰基、C ₁-C ₃烷基、C ₁-C ₃杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₃烷氧基和C ₁-C ₃卤代烷氧基，其中所述C ₁-C ₃烷基、C ₁-C ₃杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₃烷氧基和C ₁-C ₃卤代烷氧基各自任选地被一个或多个R ₈取代； If present, each R ₅ is independently selected from hydrogen, halogen, hydroxy, amino, cyano, carbamoyl, C ₁ -C ₃ alkyl, C ₁ -C ₃ heteroalkyl, C ₃ -C ₈ ring Alkyl, C ₃ -C ₈ heterocycloalkyl, C ₁ -C ₃ alkoxy and C ₁ -C ₃ haloalkoxy, wherein said C ₁ -C ₃ alkyl, C ₁ -C ₃ heteroalkyl , C3 _- _C8cycloalkyl , C3- _{C8heterocycloalkyl} , _C1 _- _C3alkoxy , and _C1 _- C3haloalkoxy are each optionally substituted with one or more _R8 ;

若存在，每一个R ₈各自独立地选自氢、卤素、羟基、氨基和氰基； If present, each R is _{independently} selected from hydrogen, halogen, hydroxy, amino and cyano;

A和B各自独立地选自C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₆-C ₁₀芳基和C ₅-C ₁₂杂芳基； A and B are each independently selected from C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl, and C ₅ -C ₁₂ heteroaryl;

其中所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3； wherein the heteroatoms or heteroatomic groups in the heteroalkyl, heterocycloalkyl and heteroaryl groups are each independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S -, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O) NH-, the number of heteroatoms or heteroatoms in the heteroalkyl, heterocycloalkyl and heteroaryl groups is each independently selected from 1, 2 and 3;

n为1、2或3。n is 1, 2 or 3.
根据权利要求1所述的化合物，其特征在于，其为如式I-1或式I-2所示的化合物，其中The compound according to claim 1, characterized in that it is a compound represented by formula I-1 or formula I-2, wherein

X ⁵、X ⁶和X ⁷各自独立地选自CR ₅和N； X ⁵ , X ⁶ and X ⁷ are each independently selected from CR ₅ and N;

X ⁸、R ₁、R ₂、R ₃、R ₄、R ₅、R ₆、B和n如权利要求1中所定义。 X ⁸ , R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , B and n are as defined in claim 1 .
根据权利要求2所述的化合物，其特征在于，The compound of claim 2, wherein

X ⁵、X ⁶和X ⁷各自独立地选自CR ₅和N； X ⁵ , X ⁶ and X ⁷ are each independently selected from CR ₅ and N;

X ⁸为N或NR ₅； X ⁸ is N or NR ₅ ;

R ₁为氢、氨基、氧代基或C ₁-C ₈烷基，其中所述C ₁-C ₈烷基任选地被一个或多个R ₅取代； R ₁ is hydrogen, amino, oxo or C ₁ -C ₈ alkyl, wherein said C ₁ -C ₈ alkyl is optionally substituted with one or more R ₅ ;

R ₂为氢； R ₂ is hydrogen;

R ₃为氢或羟基； R ₃ is hydrogen or hydroxyl;

每一个R ₄各自独立地选自氢、卤素和C ₁-C ₈烷基，其中所述C ₁-C ₈烷基任选地被一个或多个R ₅取代； Each R ₄ is independently selected from hydrogen, halogen and C ₁ -C ₈ alkyl, wherein said C ₁ -C ₈ alkyl is optionally substituted with one or more R ₅ ;

R ₆为氢； R ₆ is hydrogen;

若存在，每一个R ₅各自独立地选自氢、氨基、氨基甲酰基、C ₁-C ₃烷基和C ₁-C ₃杂烷基，其中所述C ₁-C ₃烷基、C ₁-C ₃杂烷基、C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₁-C ₃烷氧基和C ₁-C ₃卤代烷氧基各自任选地被一个或多个R ₈取代；优选地，若存在，每一个R ₅各自独立地选自氢、氨基、氨基甲酰基和C ₁-C ₃烷基，其中所述C ₁-C ₃烷基任选地被一个或多个R ₈取代； If present, each R ₅ is independently selected from hydrogen, amino, carbamoyl, C ₁ -C ₃ alkyl and C ₁ -C ₃ heteroalkyl, wherein said C ₁ -C ₃ alkyl, C ₁ _-C3 -heteroalkyl, C3 _- _C8cycloalkyl , C3- _{C8heterocycloalkyl} , _C1 _- _C3alkoxy , and _C1 _- C3haloalkoxy are each optionally replaced by one or Multiple R ₈ substitutions; preferably, if present, each R ₅ is independently selected from hydrogen, amino, carbamoyl, and C ₁ -C ₃ alkyl, wherein said C ₁ -C ₃ alkyl is optionally is substituted by one or more R ₈ ;

若存在，每一个R ₈各自独立地选自氢和羟基； If present, each R is _{independently} selected from hydrogen and hydroxyl;

B为C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₆-C ₁₀芳基和C ₅-C ₁₂杂芳基； B is C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl and C ₅ -C ₁₂ heteroaryl;

其中所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂烷基、杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3；优选地，所述杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3； wherein the heteroatoms or heteroatomic groups in the heteroalkyl, heterocycloalkyl and heteroaryl groups are each independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S -, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O) NH-, the number of heteroatoms or heteroatom groups in the heteroalkyl, heterocycloalkyl and heteroaryl groups is each independently selected from 1, 2 and 3; preferably, the heterocycloalkyl and heteroaryl groups The heteroatoms or heteroatomic groups in are independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S-, -C(=O)O-, -C(= O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O)NH-, among the heterocycloalkyl and heteroaryl the number of heteroatoms or heteroatomic groups is each independently selected from 1, 2 and 3;

n为1、2或3。n is 1, 2 or 3.
根据权利要求2或3所述的化合物，其特征在于，The compound according to claim 2 or 3, characterized in that,

结构中的
片段选自下列片段中的任意一种： in structure
Fragments are selected from any of the following fragments:

优选地，结构中的
片段选自下列片段中的任意一种： Preferably, in the structure
Fragments are selected from any of the following fragments:

或者优选地，结构中的
片段选自下列片段中的任意一种： Or preferably, in the structure
Fragments are selected from any of the following fragments:
根据权利要求1所述的化合物，其特征在于，其为如式I-3或式I-4所示的化合物，其中The compound according to claim 1, characterized in that it is a compound represented by formula I-3 or formula I-4, wherein

X ¹、X ²和X ³各自独立地选自CR ₅或N，X ⁵和X ⁶各自独立地选自C和N，X ⁷为CR ₅或N； X ¹ , X ² and X ³ are each independently selected from CR ₅ or N, X ⁵ and X ⁶ are each independently selected from C and N, and X ⁷ is CR ₅ or N;

X ⁸、R ₁、R ₂、R ₃、R ₄、R ₅、R ₆、B和n如权利要求1中所定义。 X ⁸ , R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , B and n are as defined in claim 1 .
根据权利要求5所述的化合物，其特征在于，The compound of claim 5, wherein

X ¹、X ²和X ³各自独立地选自CR ₅或N，X ⁵和X ⁶各自独立地选自C和N，X ⁷为CR ₅或N； X ¹ , X ² and X ³ are each independently selected from CR ₅ or N, X ⁵ and X ⁶ are each independently selected from C and N, and X ⁷ is CR ₅ or N;

X ⁸为N或NR ₅； X ⁸ is N or NR ₅ ;

R ₁为氢或C ₁-C ₈烷基，其中所述C ₁-C ₈烷基任选地被一个或多个R ₅取代； R ₁ is hydrogen or C ₁ -C ₈ alkyl, wherein said C ₁ -C ₈ alkyl is optionally substituted with one or more R ₅ ;

R ₂为氢； R ₂ is hydrogen;

R ₃氢或羟基； R ₃ hydrogen or hydroxyl;

每一个R ₄各自独立地选自氢、卤素和C ₁-C ₈烷基，其中所述C ₁-C ₈烷基任选地被一个或多个R ₅取代； Each R ₄ is independently selected from hydrogen, halogen and C ₁ -C ₈ alkyl, wherein said C ₁ -C ₈ alkyl is optionally substituted with one or more R ₅ ;

R ₆为氢； R ₆ is hydrogen;

若存在，每一个R ₅各自独立地选自氢和C ₁-C ₃烷基； If present, each R ₅ is independently selected from hydrogen and C ₁ -C ₃ alkyl;

B为C ₃-C ₈环烷基、C ₃-C ₈杂环烷基、C ₆-C ₁₀芳基和C ₅-C ₁₂杂芳基； B is C ₃ -C ₈ cycloalkyl, C ₃ -C ₈ heterocycloalkyl, C ₆ -C ₁₀ aryl and C ₅ -C ₁₂ heteroaryl;

其中所述杂环烷基和杂芳基中的杂原子或杂原子团各自独立地选自-C(＝O)NH-、-NH-、-N＝、-O-、-S-、-C(＝O)O-、-C(＝O)-、-C(＝S)-、-S(＝O)-、-S(＝O) ₂-和-NHC(＝O)NH-，所述杂环烷基和杂芳基中的杂原子或杂原子团的数目各自独立地选自1、2和3； wherein the heteroatoms or heteroatomic groups in the heterocycloalkyl and heteroaryl groups are each independently selected from -C(=O)NH-, -NH-, -N=, -O-, -S-, -C (=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O) ₂ - and -NHC(=O)NH-, so The number of heteroatoms or heteroatomic groups in the heterocycloalkyl and heteroaryl groups is each independently selected from 1, 2 and 3;

n为1、2或3。n is 1, 2 or 3.
根据权利要求5或6所述的化合物，其特征在于，The compound according to claim 5 or 6, characterized in that,

结构中的
片段选自下列片段中的任意一种： in structure
Fragments are selected from any of the following fragments:

优选地，结构中的
片段选自下列片段中的任意一种： Preferably, in the structure
Fragments are selected from any of the following fragments:
根据权利要求2至7中任一项所述的化合物，其特征在于，The compound of any one of claims 2 to 7, wherein

结构中的
片段选自下列片段中的任意一种： in structure
Fragments are selected from any of the following fragments:

优选地，结构中的
片段选自下列片段中的任意一种： Preferably, in the structure
Fragments are selected from any of the following fragments:
下列化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药：The following compounds or their pharmaceutically acceptable salts, hydrates, solvates, stereoisomers, tautomers, metabolites or prodrugs:
根据权利要求1所述的化合物的制备方法，其包括下列步骤：The preparation method of compound according to claim 1, it comprises the following steps:

1)化合物A与化合物B反应，得到化合物C；1) compound A reacts with compound B to obtain compound C;

2)化合物C与化合物D反应，得到化合物E；和2) compound C reacts with compound D to obtain compound E; and

3)化合物E与化合物F反应，得到目标产物；3) compound E reacts with compound F to obtain the target product;

其中in

LG ₁和LG ₂各自独立地选自氯和溴，LG ₃为氯、溴或羟基； LG ₁ and LG ₂ are each independently selected from chlorine and bromine, and LG ₃ is chlorine, bromine or hydroxyl;

X ¹、X ²、X ³、X ⁴、X ⁵、X ⁶、X ⁷、X ⁸、R ₁、R ₂、R ₃、R ₄、R ₆、R ₇、A、B和n如权利要求1中所定义。 X ¹ , X ² , X ³ , X ⁴ , X ⁵ , X ⁶ , X ⁷ , X ⁸ , R ₁ , R ₂ , R ₃ , R ₄ , R ₆ , R ₇ , A, B and n as claimed as defined in 1.
一种药物组合物，其包含根据权利要求1至9中任一项所述的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药，和至少一种药学上可接受的辅料。A pharmaceutical composition comprising a compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite thereof or a prodrug, and at least one pharmaceutically acceptable excipient.
根据权利要求1至9中任一项所述的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药或者根据权利要求11所述的药物组合物，其用作SHP2抑制剂或者用于预防和/或治疗与SHP2活性异常相关的疾病或病症；A compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug thereof or according to claim 11 Described pharmaceutical composition, it is used as SHP2 inhibitor or is used for preventing and/or treating the disease or condition associated with abnormal SHP2 activity;

优选地，所述与SHP2活性异常相关的疾病或病症选自努南综合征、豹皮综合征、白血病、成神经细胞瘤、黑色素瘤、乳腺癌、食道癌、肺癌、结肠癌、头颈部肿瘤、胃癌、间变性大细胞淋巴瘤和成胶质细胞瘤，优选非小细胞肺癌、食管癌和头颈部肿瘤。Preferably, the disease or condition associated with abnormal SHP2 activity is selected from Noonan syndrome, leopard skin syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, lung cancer, colon cancer, head and neck cancer Tumors, gastric cancer, anaplastic large cell lymphoma and glioblastoma, preferably non-small cell lung cancer, esophageal cancer and head and neck tumors.
一种用于预防和/或治疗与SHP2活性异常相关的疾病或病症的方法，其包括将预防和/或治疗有效量的根据权利要求1至9中任一项所述的化合物或其药学上可接受的盐、水合物、溶剂化物、立体异构体、互变异构体、代谢产物或前药或者根据权利要求11所述的药物组合物施用于对其有需要的个体；A method for preventing and/or treating a disease or condition related to abnormal SHP2 activity, comprising the compound of any one of claims 1 to 9 or a pharmaceutically effective amount thereof for preventing and/or treating an acceptable salt, hydrate, solvate, stereoisomer, tautomer, metabolite or prodrug or the pharmaceutical composition of claim 11 for administration to an individual in need thereof;

优选地，所述与SHP2活性异常相关的疾病或病症选自努南综合征、豹皮综合征、白血病、成神经细胞瘤、黑色素瘤、乳腺癌、食道癌、肺癌、结肠癌、头颈部肿瘤、胃癌、间变性大细胞淋巴瘤和成胶质细胞瘤，优选非小细胞肺癌、食管癌和头颈部肿瘤。Preferably, the disease or condition associated with abnormal SHP2 activity is selected from Noonan syndrome, leopard skin syndrome, leukemia, neuroblastoma, melanoma, breast cancer, esophageal cancer, lung cancer, colon cancer, head and neck cancer Tumors, gastric cancer, anaplastic large cell lymphoma and glioblastoma, preferably non-small cell lung cancer, esophageal cancer and head and neck tumors.