TW202043212A - Shp2 inhibitor and application thereof - Google Patents

Abstract

Provided is a compound for use as an inhibitor of Src homology region 2-containing protein tyrosine phosphatase 2 (SHP2) (as represented by formula I), and pharmaceutical composition and preparation method thereof, and use thereof in the treatment of SHP2-mediated diseases. The compound works by means of participating in various processes, such as regulating cell proliferation, apoptosis, migration, and neovascularization.

Description

SHP2 inhibitor and its application

The present invention relates to a series of compounds as inhibitors of Src homologyregion 2-containing protein tyrosine phosphatase 2, SHP2, and a preparation method and pharmaceutical composition thereof. The present invention also relates to the use of the above-mentioned compound or its pharmaceutical composition in the treatment of SHP2-mediated diseases.

Src homologyregion 2-containing protein tyrosine phosphatase 2 (Src homologyregion 2-containing protein tyrosine phosphatase 2, SHP2) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene. PTPN11 is the first The discovered proto-oncogene encoding tyrosine kinase (Chan RJ et al. PTPN11 is the first identified proto-oncogene that encodes a tyrosine phosphatase. Blood, 2007, 109:862-867), and the encoded SHP2 protein contains N-terminal SHP2 domain (N-SHP2), C-terminal SHP2 domain (C-SHP2), protein phosphatase catalytic domain (PTP), two C-terminal tyrosine residues (Y542 and Y580) and one proline-rich Acid (Pro) motif.

In recent years, research mainly believes that the Ras/ERK pathway is the most important signal transduction pathway for SHP2, and its mechanism (Dance M et al. The molecular functions of Shp2 in the RAS/mitogen-activated protein kinase (ERK1/2) pathway. Cell Signal, 2008, 20:453-459) roughly: After the activation of growth factor receptors, its tyrosine residues undergo autophosphorylation, which are Grb2 and SHP2 (adapter proteins containing SH2 domains) phosphotyrosine The binding area SH2 provides a stopping point. The binding of Grb2 to phosphorylated growth factor receptors results in the accumulation of SOS protein in the cell membrane. As a kind of guanine nucleotide exchange factor (GEF), SOS can catalyze the conversion of membrane-bound protein Ras from inactive Ras-GDP to active Ras-GTP. Ras-GTP further connects with the downstream signaling system, activates Ser/Thr kinase Raf1, etc., and then activates ERK under the action of the regulatory kinase MEK. After activation, ERK directly acts on target molecules in the cytoplasm or transfers to the nucleus to regulate genes Transcription to make cells proliferate or differentiate This process may also be affected by SHP2 binding protein and substrate (SHP substrate-1, SHPS-1), Ras-GTPase activating protein (Ras-GAP) and other members of Src.

SHP2 protein not only regulates the Ras/ERK signaling pathway, it is also reported to regulate multiple signaling pathways such as JAK-STAT3, NF-κB, PI3K/Akt, RHO, and NFAT, thereby regulating cell proliferation, differentiation, migration, and apoptosis. Learn function.

SHP2 has been proved to be related to many diseases. Tartaglia et al. (Tartaglia M et al. Mutations in PTPN11, encoding the protein tyrosine phosphatase SHP-2, cause Noonan syndrome. Nat Genet, 2001, 29:465-468) found that about 50% Patients with Southern syndrome have missense mutations of PTPN11. In addition, studies have found that PTPN11 mutation is an important cause of JMML and a variety of leukemias (Tartaglia M et al. Nat Genet, 2003, 34: 148-150; Loh ML et al. Blood, 2004, 103: 2325-2331; Tartaglia M et al. al. Br J Haematol, 2005, 129:333-339; Xu R et al. Blood, 2005, 106:3142-3149.). With the deepening of research on PTPN11/SHP2, it has been found that it is related to the occurrence of lung cancer, gastric cancer, colon cancer, melanoma, thyroid cancer and other cancers (Tang Chunlan et al. Chinese Journal of Lung Cancer, 2010, 13: 98-101 ; Higuchi M et al. Cancer Sci, 2004, 95:442-447; Bentires-Al j M et al. Cancer Res, 2004, 64:8816-8820; Martinelli S et al. Cancer Genet Cytogenet, 2006, 166:124 -129.).

Therefore, SHP2 inhibitors have received more and more attention as potential treatments. There are a variety of SHP2 inhibitors currently under development. Novartis's TNO155 entered phase I clinical trials for the treatment of solid tumors in 2017. JAB-3068, designed and developed by Jacos, was officially approved by the US FDA for clinical trials of new drugs in January 2018. The RMC-4630 developed by Revolution undergoes its first human clinical trial in the second half of 2018. At present, this target has not yet been marketed at home and abroad. Therefore, the development of small molecule drugs that can target and inhibit SHP2 activity and provide patients with safer and more effective SHP2 inhibitors have important research significance.

式I 其中， R₁ 任意地選自氨基、-C(O)-R_a 、-C≡N 、羥基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基； R_a 任意地選自氨基、-NH-OH、C_1-3 烷基； R₂ 任意地選自氫、C_1-4 烷基或含取代基的C_1-4 烷基； R₃ 任意地選自氫、鹵素、氨基、-C(O)NH₂ 、-C≡N、羥基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基或含取代基的C_1-8 烷氧基； R₄ 任意地選自氫、鹵素、氨基、醯胺基、-C≡N、羧基、羥基、羥甲基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_2-8 烯基、含取代基的C_2-8 烯基、C_2-8 炔基或含取代基的C_2-8 炔基； A₁ 任意地選自CR₅ 或N； A₂ 任意地選自CR₆ 或N； A₃ 任意地選自CR₇ 或N； A₄ 任意地選自CR₈ 或N； U任意地選自C(R₉ )₂ 、O或NR₁₀ ； 其中，R₅ 、R₆ 、R₇ 、R₈ 、R₉ 或R₁₀ 獨立地選自氫、羥基、鹵素、氨基、含取代基的氨基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_2-8 烯基、含取代基的C_2-8 烯基、C_2-8 炔基、含取代基的C_2-8 炔基或C_5-6 雜環基；或者， R₅ 和R₆ 與他們連接的碳原子共同形成5到6員芳基或5到6員雜環基； L選自S； 環A任意地選自C_6-10 芳基或C_5-10 雜芳基，所述C_5-10 雜芳基含有一個或兩個N或S雜原子； Rx任意地選自氫、羥基、鹵素、氰基、氨基、含取代基的氨基、磺醯基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_3-8 環烷基或含取代基的C_3-8 環烷基； n為0、1、2、3或4。The present invention relates to a compound used as an inhibitor of protein tyrosine phosphatase 2 (SHP2) containing Src homology region 2. The compound of the present invention has the general structure shown in formula I or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,

Formula I wherein, R ₁ optionally selected from _{amino, -C (O) -R a,} -C≡N, hydroxy, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 Alkoxy, C _1-8 alkoxy containing substituents; R _{a is} optionally selected from amino, -NH-OH, C _1-3 alkyl; R _{2 is} optionally selected from hydrogen, C _1-4 alkyl Or C _1-4 alkyl containing substituents; R _{3 is} arbitrarily selected from hydrogen, halogen, amino, -C(O)NH ₂ , -C≡N, hydroxy, C _1-8 alkyl, substituents containing C _1-8 alkyl, C _1-8 alkoxy or substituted C _1-8 alkoxy; R _{4 is} optionally selected from hydrogen, halogen, amino, amide, -C≡N, carboxy, Hydroxy, hydroxymethyl, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _2-8 alkenyl , C _2-8 alkenyl group having a substituent, C _2-8 alkynyl or C _2-8 alkynyl group-containing substituent group; a ₁ is selected from optionally CR ₅ or N; a ₂ is selected from CR ₆ or optionally N; A _{3 is} arbitrarily selected from CR ₇ or N; A _{4 is} arbitrarily selected from CR ₈ or N; U is arbitrarily selected from C(R ₉ ) ₂ , O or NR ₁₀ ; wherein, R ₅ , R ₆ , R ₇ , R ₈ , R ₉ or R _{10 are} independently selected from hydrogen, hydroxy, halogen, amino, substituted amino, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy group, a substituted C _1-8 alkoxy group, C _2-8 alkenyl group, a substituted group containing C _2-8 alkenyl, C _2-8 alkynyl group, a substituted group containing C _2-8 alkynyl Or C _5-6 heterocyclic group; or, R ₅ and R ₆ and the carbon atom to which they are attached together form a 5- to 6-membered aryl group or a 5- to 6-membered heterocyclic group; L is selected from S; ring A is optionally selected From C _6-10 aryl or C _5-10 heteroaryl, the C _5-10 heteroaryl contains one or two N or S heteroatoms; Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, Amino, substituted amino, sulfonyl, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _3-8 cycloalkyl or substituted C _3-8 cycloalkyl; n is 0, 1, 2, 3 or 4.

In some embodiments, R ₁ in Formula I optionally selected from _{amino, -C (O) NH 2,} -C≡N, C 1-3 alkyl group, a substituted C _1-3 alkyl group or a C _{1 -3} Alkoxy.

、

、

、

、

或

。In some embodiments, R ₁ in formula I is arbitrarily selected from amino, -C(O)NH ₂ , -C≡N, methyl substituted by hydroxy,

,

,

,

,

or

.

In some embodiments, R ₂ in Formula I is hydrogen.

In some embodiments, R ₃ of Formula I is optionally selected from hydrogen, halo, C _1-3 alkyl or an optionally substituted C _1-3 alkyl.

In some embodiments, R ₃ in Formula I is arbitrarily selected from hydrogen, chlorine, or methyl.

In some embodiments, R ₄ in Formula I is arbitrarily selected from hydrogen, halogen, C _1-8 alkyl or C _1-8 alkoxy containing substituents.

In some embodiments, R ₄ in Formula I is arbitrarily selected from hydrogen, F, Cl, -CHF ₂ , CF ₃ or -O-CH ₃ .

In some embodiments, A ₁ in formula I is CR ₅ or N, wherein R _{5 is} selected from hydrogen, halogen, or halogen substituted C _1-3 alkyl.

In some embodiments, A ₁ in formula I is CR ₅ or N, wherein R _{5 is} selected from hydrogen, Cl or trifluoromethyl.

In some embodiments, A ₂ in Formula I is CR ₆ or N, wherein R _{6 is} selected from hydrogen, hydroxyl, halogen, amino, or C _1-8 alkoxy.

In some embodiments, A ₂ in Formula I is CR ₆ or N, wherein R _{6 is} selected from hydrogen, OH, F, Cl, amino, or -O-CH ₃ .

In some embodiments, A ₃ in Formula I is CR ₇ or N, wherein R ₇ is hydrogen or halogen.

In some embodiments, A ₄ in Formula I is CR ₈ or N, wherein R _{8 is} selected from hydrogen, halogen, amino, substituted amino, C _1-3 alkoxy, or C _5-6 heterocyclyl.

、

或甲氧基。In some embodiments, A ₄ in formula I is CR ₈ or N, wherein R _{8 is} selected from hydrogen, F, Cl, amino, -NHCH ₃ , -N(CH ₃ ) ₂ ,

,

Or methoxy.

In some embodiments, U in Formula I is CH ₂ or O.

In some embodiments, ring A in formula I is arbitrarily selected from phenyl or C _5-6 heteroaryl, and the C _5-6 heteroaryl contains one or two N or S heteroatoms.

In some embodiments, Rx in Formula I is arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C _1-3 alkyl, halogen-substituted C _1-3 alkyl, or C _1-3 alkoxy.

In some embodiments, Rx in Formula I is arbitrarily selected from hydrogen, OH, F, Cl, Br, -CN, trifluoromethyl, or methoxy.

選自

、

、

、

、

、

、

、

、

、

、

或

。In some embodiments, the formula I

Selected from

,

,

,

,

,

,

,

,

,

,

or

.

式II 其中， R₃ 任意地選自氫、鹵素、氨基、-C(O)NH₂ 、-C≡N、羥基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基或含取代基的C_1-8 烷氧基； A₁ 任意地選自CR₅ 或N； U任意地選自C(R₉ )₂ 、O或NR₁₀ ； 其中，R₅ 、R₉ 或R₁₀ 獨立地選自氫、羥基、鹵素、氨基、含取代基的氨基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_2-8 烯基、含取代基的C_2-8 烯基、C_2-8 炔基或含取代基的C_2-8 炔基或C_5-6 雜環；或者， L選自S； R_x 任意地選自氫、羥基、鹵素、氰基、氨基、含取代基的氨基、磺醯基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_3-8 環烷基或含取代基的C_3-8 環烷基； n為0、1、2、3或4。The present invention further provides some preferred technical solutions of the compound represented by formula I. For example, the compound of the present invention has the general structure shown in Formula II or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,

Formula II wherein, R ₃ is optionally selected from hydrogen, halogen, _{amino, -C (O) NH 2,} -C≡N, hydroxy, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 alkoxy or C _1-8 alkoxy containing substituents; A _{1 is} optionally selected from CR ₅ or N; U is optionally selected from C(R ₉ ) ₂ , O or NR ₁₀ ; wherein, R _5. R ₉ or R _{10 are} independently selected from hydrogen, hydroxy, halogen, amino, substituted amino, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy containing substituted C _1-8 alkoxy group, C _2-8 alkenyl group, a substituted group containing C _2-8 alkenyl, C _2-8 alkynyl group or a substituted group containing C _2-8 alkynyl or C _5-6 heterocycle; or, L is selected from S; R _{x is} arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _3-8 cycloalkyl or substituted C _3-8 cycloalkyl; n is 0, 1, 2, 3, or 4.

In some embodiments, R ₃ in Formula II is arbitrarily selected from hydrogen or C _1-3 alkyl.

In some embodiments, R ₃ in Formula II is arbitrarily selected from hydrogen or methyl.

In some embodiments, A ₁ in formula II is CR ₅ or N, wherein R _{5 is} selected from halogen.

In some embodiments, A ₁ in Formula II is CR ₅ , wherein R _{5 is} selected from F or Cl.

In some embodiments, A ₁ in Formula II is N.

In some embodiments, U in Formula II is O or CH ₂ .

In some embodiments, R _x in Formula II is arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C _1-3 alkyl, halogen-substituted C _1-3 alkyl, or C _1-3 alkoxy.

In some embodiments, R _x in Formula II is arbitrarily selected from hydrogen, OH, CN, Cl, F, Br, trifluoromethyl, or methoxy.

In some embodiments, n in Formula II is 0, 1, or 2.

式III R₃ 任意地選自氫、鹵素、氨基、-C(O)NH₂ 、-C≡N、羥基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基或含取代基的C_1-8 烷氧基； R₄ 任意地選自氫、鹵素、氨基、醯胺基、-C≡N、羧基、羥基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_2-8 烯基、含取代基的C_2-8 烯基、C_2-8 炔基或含取代基的C_2-8 炔基； A₁ 任意地選自CR₅ 或N； A₂ 任意地選自CR₆ 或N； A₃ 任意地選自CR₇ 或N； U任意地選自C(R₉ )₂ 、O或NR₁₀ ； 其中，R₅ 、R₆ 、R₇ 、R₈ 、R₉ 或R₁₀ 獨立地選自氫、羥基、鹵素、氨基、含取代基的氨基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_2-8 烯基、含取代基的C_2-8 烯基、C_2-8 炔基、含取代基的C_2-8 炔基或C_5-6 雜環基； L選自S； 環A任意地選自C_6-10 芳基或C_5-10 雜芳基，所述C_5-10 雜芳基含有一個或兩個N或S雜原子； Rx任意地選自氫、羥基、鹵素、氰基、氨基、含取代基的氨基、磺醯基、C_1-8 烷基、含取代基的C_1-8 烷基、C_1-8 烷氧基、含取代基的C_1-8 烷氧基、C_3-8 環烷基或含取代基的C_3-8 環烷基； n為0、1、2、3或4。The present invention further provides some preferred technical solutions of the compound represented by formula I. For example, the compound of the present invention has the general structure shown in Formula III or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,

Optionally the formula III R ₃ is selected from hydrogen, halogen, _{amino, -C (O) NH 2,} -C≡N, hydroxy, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _{l- 8} Alkoxy or C _1-8 alkoxy containing substituents; R _{4 is} optionally selected from hydrogen, halogen, amino, amide, -C≡N, carboxyl, hydroxyl, C _1-8 alkyl, containing Substituent C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _2-8 alkenyl, substituted C _2-8 alkenyl, C _{2 -8} alkynyl or C _2-8 alkynyl containing substituents; A _{1 is} optionally selected from CR ₅ or N; A _{2 is} optionally selected from CR ₆ or N; A _{3 is} optionally selected from CR ₇ or N; U Arbitrarily selected from C(R ₉ ) ₂ , O or NR ₁₀ ; wherein R ₅ , R ₆ , R ₇ , R ₈ , R ₉ or R _{10 are} independently selected from hydrogen, hydroxyl, halogen, amino, and substituents amino, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 alkoxy, group containing C _1-8 alkoxy substituent, C _2-8 alkenyl group, a substituted C _2-8 alkenyl, C _2-8 alkynyl, C _2-8 alkynyl or C _5-6 heterocyclic group containing substituents; L is selected from S; ring A is optionally selected from C _6-10 Aryl or C _5-10 heteroaryl, the C _5-10 heteroaryl contains one or two N or S heteroatoms; Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, and substituents amino, sulfo acyl, C _1-8 alkyl, C _1-8 alkyl-containing substituent group, a C _1-8 alkoxy group, a substituted C _1-8 alkoxy group, C _3-8 cycloalkyl Alkyl group or C _3-8 cycloalkyl group containing substituents; n is 0, 1, 2, 3 or 4.

In some embodiments, R ₃ in Formula III is selected from hydrogen.

In some embodiments, R ₄ in Formula III is arbitrarily selected from hydrogen or Cl.

In some embodiments, A ₁ in Formula III is CR ₅ or N, wherein R _{5 is} selected from hydrogen, Cl or trifluoromethyl.

、或-O-CH₃ 。In some embodiments, A ₂ in formula III is CR ₆ or N, wherein R _{6 is} selected from Cl, amino,

, Or -O-CH ₃ .

In some embodiments, A ₃ in Formula III is CR ₇ or N, wherein R ₇ is hydrogen.

In some embodiments, U in Formula III is CH ₂ .

In some embodiments, ring A in formula III is arbitrarily selected from phenyl or C _5-6 heteroaryl, and the C _5-6 heteroaryl contains one or two N or S heteroatoms.

In some embodiments, Rx in Formula III is arbitrarily selected from hydrogen, Cl, Br, trifluoromethyl, or methoxy.

選自

、

、

、

、

、

、

、

。In some embodiments, the formula III

Selected from

,

,

,

,

,

,

,

.

The present invention further provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 2) ( S )-(3-(1-amino-1,3-dihydrospiro[indene-2, 4'-Pepidine] -1'-yl)-6-((2,3-dichlorophenyl)thio)-5-methylpyrazin-2-yl)methanol; 3)( S )-3 -(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)sulfanyl ) Pyrazine-2-carbonitrile; 4) ( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-( (2,3-Dichlorophenyl)thio)pyrazine-2-carboxamide; 5)( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'- Piperidine] -1'-yl)-6-((2-amino-3-chloropyridin-4-yl)sulfanyl)pyrazine-2-carboxamide; 7)( S )-3-(1- Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methoxypyridin-4-yl)thio)pyridine Oxazine-2-carboxamide; 8)( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(( 2-(Trifluoromethyl)pyridin-3-yl)thio)pyrazine-2-carboxamide; 9)( S )-3-(1-amino-6-methoxy-1,3-di Hydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl)pyridin-3-yl)thio)pyrazine-2-methanamide; 10) ( S )-6-((2-amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-bromo-1,3-dihydrospiro(indene-2, 4'-Pepidine]-1'-yl)pyrazine-2-carboxamide; 11)( S )-6-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3- (1-Amino-4-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide; 12)( S )- 6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-呱(Pyridin)-1'-yl)pyrazine-2-carboxamide; 13)( S )-6-((2-amino-3-chloropyridin-4-yl)thio)-3-(1-amino -5,6-Dimethyl-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide; 14)( S )-6- ((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-(trifluoromethyl)-1,3-dihydrospiro(indene-2,4'- Pyridine]-1'-yl)pyrazine-2-carboxamide; 15)( S )-6-((2-amino-3-chloropyridin-4-yl)thio)-3-(1- Amino-6-chloro-1, 3-Dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide; 16)( S )-3-(1-amino-1,3-dihydro Spiro[indene-2,4'-pyridine]-1'-yl)-6-((6-amino-3-chloropyridin-2-yl)thio)pyrazine-2-methanamide; 17) ( S )-3-(5-Amino-2-methoxy-5,7-dihydrospiro[cyclopentadiene[ b ]pyridine-6,4'-piperidine]-1'-yl)-6 -((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide; 18) ( S )-3-(4-amino-2-chloro-4,6-di Hydrospiro[cyclopenta[ d ]thiazole-5,4'-pyridine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazine -2-methylamide; 19) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(( 2-Fluoro-3-methoxyphenyl)thio)pyrazin-2-yl)methanol; 20) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2, 4'-Pepidine]-1'-yl)-6-((2-methoxypyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 21) (S)-(3-( 1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)sulfanyl)pyridine Azin-2-yl)methanol; 22) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-( (2-Aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 23) (S)-1-(3-(1-amino-1,3-dihydrospiro[indene-2, 4'-Pepidine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)ethan-1-one; 24) (S )-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-((1-methyl -1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol; 25) (S)-(3-(1-amino-1,3-di Hydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-(trifluoromethyl)pyridin-4-yl)thio)pyrazin-2-yl)methanol; 26) (S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-(trifluoromethyl)-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol; 27) 1-(3-((S)-1-amino-1,3-dihydrospiro [Indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)ethan-1-ol ; 28) (S)-(3- (1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((5-chloro-2-((1-methyl-1H-pyridine) (Azol-5-yl)amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol; 29) (S)-(3-(1-amino-1,3-dihydrospiro[indene- 2,4'-Pepidine]-1'-yl)-6-((2-(dimethylamino)-3-fluoropyridin-4-yl)thio)pyrazin-2-yl)methanol; 30 ) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-fluoro-2-(methyl Amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol; 31) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-呱Pyridin]-1'-yl)-6-((2-(difluoromethyl)pyridin-3-yl)thio)pyrazin-2-yl)methyl; 32) (S)-(3-( 1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-(dimethylamino)pyridin-4-yl )Thio)pyrazin-2-yl)methyl; 33) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'- Yl)-6-((3-fluoro-2-methoxyphenyl)thio)pyrazin-2-yl)methanol; 34) (S)-(3-(1-amino-1,3-di Hydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-5-fluoro-2-methoxyphenyl)thio)pyrazin-2-yl) Methanol; 35) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(quinolin-4-yl Thio)pyrazin-2-yl)methanol; 36) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl) -6-((2-(Trifluoromethyl)pyridin-3-yl)thio)pyrazin-2-yl)methanol; 37) (S)-(3-(1-amino-1,3-di Hydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorophenyl)thio)pyrazin-2-yl)methanol; 38) (S) -(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl)pyridine-3- Yl)thio)pyrazin-2-yl)methanol; 39) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'- Yl)-6-((2-amino-3-fluoropyridin-4-yl)thio)pyrazin-2-yl)methyl; 40) (S)-(3-(1-amino-1,3 -Dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-chloro-2-methoxypyridin-4-yl)thio)pyrazin-2-yl ) Methanol; 41) (S)-(6-((2-amino-3-chloro Pyridin-4-yl)thio)-3-(1-amino-6-fluoro-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)pyrazine-2- Group) methanol; 42) (S)-(6-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-methoxy-1,3-di Hydrospiro[inden-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol; 43) (S)-(6-((2-amino-3-chloropyridine-4- Yl)thio)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2- Yl)methanol; 44) (R)-(6-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(3-amino-3H-spiro[benzofuran-2,4 '-Pepidine]-1'-yl)pyrazin-2-yl)methyl; 45) (S)-1-amino-1'-(5-((2-amino-3-chloropyridine-4- (Yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-4-carbonitrile; 46) (S)- (6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2,4'-piperidine ]-1'-yl)pyrazin-2-yl)methanol; 47) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1 '-Yl)-6-(pyridin-4-ylthio)pyrazin-2-yl)methanol; 48) (S)-(6-((2-amino-3-chloropyridin-4-yl)sulfur Yl)-3-(5-amino-5,7-dihydrospiro[cyclopenteno[b]pyridine-6,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl ; 49) (S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-(trifluoromethyl)-1,3-bis Hydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol; 50) (S)-1-amino-1'-(5-((2-amino- 3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-6-ol ; 51) (S)-4-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl) Pyrazin-2-yl)thio)pyrimidin-2-ol; 52) (S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio) -3-(hydroxymethyl)pyrazin-2-yl)-7-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-4-ol; 53) (S)-( 3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dihydro-[1,4] dioxy Heterocyclic hexano[2,3-b]pyridine-8- (Amino)thio)pyrazin-2-yl)methanol; 54) (S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3 -(Hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-6-nitrile; 55) (S)-1-amino-1'- (5-((2-Amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-4-fluoro-1,3-dihydrospiro(indene -2,4'-piperidine]-7-nitrile; 56) (S)-(6-((2-amino-3-chloropyridin-4-yl)thio)-3-(1-amino-7 -(Trifluoromethyl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl; 57) (S)-1- Amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[ Indene-2,4'-piperidine]-7-ol; 58) (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'- Yl)-6-((2-amino-3-fluoropyridin-4-yl)thio)pyrazine-2-carboxamide; 59) (S)-3-(1-amino-1,3-di Hydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-chloro-2-((1-methyl-1H-pyrazol-5-yl)amino)pyridine- 4-yl)thio)pyrazine-2-carboxamide; 60) (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-Yl)-6-((2-amino-3-chloropyridin-4-yl)thio)-N-hydroxypyrazine-2-carboxamide; 61) 1-(3-((S)-1 -Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazine -2-yl)-2,2,2-trifluoroethane-1-ol; 62) (S)-1'-(5-((2-amino-3-chloropyridin-4-yl)thio) -3-(Methoxymethyll)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine; 63) (S)-2- (3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl) Sulfuryl)pyrazin-2-yl)propan-2-ol; 64) (S)-(3-(1-amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-Pepidine]-1'-yl)-6-((2-aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 65) (S)-(3-(1-amino- 1,3-Dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(pyrimidin-4-ylthio)pyrazin-2-yl)methanol; 66) (S) -(3-(1-Amino-5-chloro-1,3- Dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 67) (S )-(3-(1-Amino-5-fluoro-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-aminopyrimidine-4- Yl)thio)pyrazin-2-yl)methanol; 68) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'- Yl)-6-((2-(methylamino)pyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 69) (S)-(3-(1-amino-1,3- Dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidin-4-yl)sulfanyl)-5-methylpiperazine-2-yl)methanol ; Or 70) (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-4 -Yl)thio)pyrazine-2-carboxamide.

The present invention also provides a pharmaceutical composition characterized by comprising a therapeutically effective amount of at least one compound represented by formula (I) and at least one pharmaceutically acceptable excipient.

The present invention further provides a pharmaceutical composition, characterized in that the mass percentage of the therapeutically effective amount of at least one compound represented by formula (I) and pharmaceutically acceptable excipients is 0.0001:1-10.

The invention provides the application of the compound or pharmaceutical composition represented by the structural formula (I) in the preparation of medicines.

The present invention further provides a better technical solution for the application.

Preferably, the application is an application for preparing a medicine for treating, preventing, delaying or preventing cancer, cancer metastasis, cardiovascular disease, immune disease, fibrosis or eye disease.

Preferably, the application is an application in preparing a medicine for treating diseases mediated by SHP2. Preferably, the disease is cancer.

Preferably, the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal tumors , Lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma, pancreatic cancer or a combination thereof.

Preferably, the application is an application for preparing an SHP2 inhibitor.

The present invention also provides a method for treating and/or preventing diseases mediated by SHP2, which comprises administering a therapeutically effective amount of at least any one compound or pharmaceutical composition represented by structural formula (I) to the treatment object.

Preferably, in the above method, the SHP2-mediated disease is cancer.

The present invention also provides a method for treating cancer, which comprises administering a therapeutically effective amount of at least any one compound or pharmaceutical composition represented by structural formula (I) to the treatment object.

Preferably, the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal tumors , Lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma, pancreatic cancer or a combination thereof.

Preferably, in the above method, the treatment object is a human being.

Unless otherwise stated, the terms used in the present invention have the following meanings:

The term "alkyl" includes straight, branched or cyclic saturated alkyl groups. For example, alkyl groups include but are not limited to methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, 3 -(2-Methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, cyclopentyl, n-hexyl, 2-hexyl, 2-methylpentyl and cyclohexyl and other similar groups group. Similarly, the " _C1-8 " in "C _1-8 alkyl" refers to a straight, branched or cyclic form containing 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms Aligned groups.

"Alkenyl" and "alkynyl" include linear, branched or cyclic alkenyl and alkynyl groups. Similarly, "C _2-8 alkenyl" and "C _2-8 alkynyl" refer to a linear, branched or cyclic arrangement containing 2, 3, 4, 5, 6, 7 or 8 carbon atoms的alkenyl or alkynyl.

The term "alkoxy" refers to the oxyether form of the aforementioned linear, branched or cyclic alkyl group.

The term "aryl" refers to an unsubstituted or substituted monocyclic or polycyclic aromatic group including carbon atoms. It is preferably a monocyclic or bicyclic aromatic group of 6 to 10 members. Phenyl and naphthyl are preferred. The most preferred is phenyl.

The term "heteroaryl" refers to a monovalent heteroatom group formed by removing a hydrogen atom from a carbon atom of a parent heteroaromatic ring system. Heteroaryl includes: 5- to 7-membered aromatic, monocyclic, including at least one heteroatom selected from N, O or S, for example, 1 to 4 heteroatoms, or preferably 1 to 3 heteroatoms, The other atoms in the ring are carbon; the polyheteroaryl ring includes at least one heteroatom selected from N, O or S, for example, 1 to 4 heteroatoms, or preferably 1 to 3 heteroatoms, on the ring The other atoms are carbon, and at least one of the heteroatoms is on the aromatic ring. Particularly preferred heteroaryl groups are C _3-10 heteroaryl groups, including but not limited to pyrrolyl, furyl, thienyl, pyridyl, pyranyl, pyrazolyl, pyrimidinyl, pyridazinyl , Pyrazinyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, indolyl, benzofuranyl, benzothiazolyl, benzimidazolyl, benzopyrazolyl, Benzotriazole, azazolyl, quinolinyl, isoquinolinyl, purinyl and similar groups.

However, in any case, the heteroaryl group and the aryl group will not cross or contain each other. Therefore, according to the above definition, if at least one all-carbon aromatic ring is fused with a heterocyclic group, a heteroaryl group is obtained instead of an aryl group.

"Heterocyclic group" refers to a saturated or unsaturated but not aromatic cyclic group, and one or more of the carbon atoms (and the attached hydrogen atom) can be respectively the same or different hetero Atom and the corresponding attached hydrogen atom are replaced. Representative heteroatoms that replace carbon atoms include, but are not limited to, N, P, O, S, and Si. When a specific degree of saturation needs to be described, the terms "heterocycloalkyl" or "heterocycloalkenyl" are used respectively. Representative heterocyclic groups include, but are not limited to, epoxy compounds, imidazolidine, morpholine, piperazine, piperidine, pyrazolidine, pyrrolidine, quinuclidine, tetrahydrofuran or tetrahydropyran and similar groups . Substituent-containing heterocyclic groups also include ring systems substituted with at least one oxygen-containing (=O) or oxide (-O-) substituent, such as: piperidine-nitrogen-oxide, morpholinyl-nitrogen- Oxide, 1-oxo-1-thiomorpholinyl and 1-dioxo-1-thiomorpholinyl.

However, in any case, the heterocycloalkyl group and the cycloalkyl group will not cross or contain each other. Therefore, according to the above definition, if at least one all-carbocyclic ring is fused with a heterocycloalkyl group to form a di-, poly- or spiro-ring, it will still be defined as a heterocycloalkyl group.

In addition, if a heteroaryl group is fused with a heterocyclic group to form a di-, poly- or spiro-ring, it will be defined as a heterocyclic group instead of a heteroaryl group.

"Halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). The preferred halogen refers to fluorine, chlorine and bromine.

"Halo" refers to a fluoro, chloro, bromo or iodo group. The preferred halo group refers to fluoro and chloro.

"Substitution" means that one or more hydrogen atoms in a group are replaced by the same or different substituents. Representative substituents include, but are not limited to, halogen, amino, hydroxyl, oxo, carbonyl, cyano, -C(O)NH ₂ , alkyl, alkoxy, aryl, cycloalkyl, heterocyclic group , Heteroaryl. In some embodiments, the substituent includes but is not limited to halogen, amino, hydroxyl, cyano, methyl, -CH ₂ OH, -C(O)NH ₂ , -OCH ₃ , trifluoromethyl.

Whenever the term "alkyl" or "aryl" or its initial root appears in the name of a substituent (such as aralkyl, or dialkylamino), it shall be referred to as the aforementioned "alkyl" and "aryl""The definition defines the substituents. The specified number of carbon atoms (such as C _1-6 ) will independently represent the number of carbon atoms in an alkyl moiety or in an alkyl moiety (where the alkyl group is the initial root) in a larger substituent.

The "compound" of the present invention includes the compound represented by formula (I) and all pharmaceutically acceptable forms thereof. These pharmaceutically acceptable forms include salts, solvates, non-covalent complexes, chelates or their prodrugs, or any mixture of all the above forms.

The "pharmaceutically acceptable" refers to those that are known to be used in animals, especially those that can be used in humans.

The term "composition" in the present invention includes a product containing a specific amount of a specific component, and also includes any product obtained directly or indirectly from a specific amount of a specific component. Therefore, a pharmaceutical composition including the compound of the present invention as an active ingredient and a method for preparing the compound are the content of the present invention.

"Therapeutically effective amount" means that when a compound is administered to a subject to treat and prevent and/or inhibit at least one clinical symptom of a disease, condition, symptom, indication and/or discomfort, it is sufficient for the disease, condition, symptom, A dose that produces a certain effect for the treatment of indications or discomfort. The specific "effective therapeutic dose" can vary depending on the compound, the route of administration, the age of the patient, the weight of the patient, the type of disease or discomfort to be treated, the symptoms and severity, etc. Wherever possible, an appropriate dose may be obvious to those skilled in the art, or it may be determined by conventional experimental methods.

The compounds provided by the present invention may exist in the form of "pharmaceutically acceptable salts". In terms of pharmaceutical applications, the salt of the compound provided by the present invention refers to a non-toxic pharmaceutically acceptable salt. The form of pharmaceutically acceptable salts includes pharmaceutically acceptable acid/anion or base/cation salts. Pharmaceutically acceptable acid/anionic salts generally exist in the form of protonation of basic nitrogen with inorganic or organic acids. Typical organic or inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, perchloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, lactic acid, succinic acid, maleic acid, fumaric acid, malic acid , Tartaric acid, citric acid, α-ketoglutaric acid, hippuric acid, benzoic acid, mandelic acid, methanesulfonic acid, isethionic acid, benzenesulfonic acid, oxalic acid, pamoic acid, 2-naphthalenesulfonic acid, p-toluenesulfonic acid Acid, cyclohexamine sulfonic acid, salicylic acid, saccharinic acid or trifluoroacetic acid. Pharmaceutically acceptable base/cation salts, including, but not limited to, aluminum salts, calcium salts, chloroprocaine salts, choline, diethanolamine salts, ethylenediamine salts, lithium salts, magnesium salts, potassium salts, sodium Salt and zinc salt.

The prodrug of the compound of the present invention is included in the protection scope of the present invention. Generally, the prodrug is a functional derivative that is easily converted into the desired compound in the body. Therefore, the term "administration" involved in the treatment method provided by the present invention includes the administration of the compound disclosed in the present invention, or although it is not clearly disclosed but can be transformed into the compound disclosed in the present invention after administration to the subject to treat various diseases. . The conventional methods for selecting and preparing suitable prodrug derivatives have been described in books such as "Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985).

Obviously, the definition of any substituent or variable at a specific position in one molecule is irrelevant to the definition of any substituent or variable at a specific position in other molecules. It is easy to understand that the compounds of the present invention can be selected according to the prior art of the subject to select suitable substituents or substitution forms to provide chemically stable and easy preparation and synthesis using the prior art of the subject or the method described in the present invention.

When the compound represented by formula (I) and its pharmaceutically acceptable salt are in the form of a solvate or polymorph, the present invention includes any possible solvate and polymorph. The type of solvent that forms the solvate is not particularly limited, as long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone and similar solvents can be used.

The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound provided by the present invention is an acid, its corresponding salt can be prepared from pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper (ic and ous), iron, ferrous, lithium, magnesium, manganese (ic and ous), potassium, sodium, zinc and the like. In particular, ammonium, calcium, magnesium, potassium, and sodium salts are preferred. Non-toxic organic bases that can be derivatized into pharmaceutically acceptable salts include primary, secondary and tertiary amines, as well as cyclic amines and amines containing substituents, such as naturally occurring and synthetic amines containing substituents. Other pharmaceutically acceptable non-toxic organic bases capable of forming salts include ion exchange resins and arginine, betaine, caffeine, choline, N',N'-dibenzylethylene diamine, diethylamine, 2- Diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, reduced glucosamine, glucosamine, histidine, haamine, isopropylamine, Lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

When the compound provided by the present invention is a base, the corresponding salt can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids and organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, isethionic acid, formic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, Hydrochloric acid, lactic acid, maleic acid, malic acid, mandelic acid, α-ketoglutaric acid, hippuric acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonate Acid etc. Preferably, malic acid, citric acid, hydrobromic acid, hydrochloric acid, methanesulfonic acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid. More preferably, phosphoric acid, hydrochloric acid and malic acid. Since the compound represented by formula (I) will be used as a medicine, it is preferable to use a substantially pure form, for example, at least 60% purity, more suitably at least 75% purity, particularly suitably at least 98% purity (% is weight ratio ).

The pharmaceutical composition provided by the present invention includes a compound represented by formula (I) (or a pharmaceutically acceptable salt thereof) as an active component, a pharmaceutically acceptable excipient and other optional therapeutic components or adjuvants . Although in any given case, the most suitable way of administering the active ingredient depends on the particular subject to be administered, the nature of the subject and the severity of the disease, the pharmaceutical composition of the present invention includes oral, rectal, topical and Pharmaceutical compositions for parenteral administration (including subcutaneous administration, intramuscular injection, and intravenous administration). The pharmaceutical composition of the present invention can be conveniently prepared in a unit dosage form known in the art and prepared by any preparation method known in the pharmaceutical field.

In fact, according to conventional drug mixing technology, the compound represented by formula (I) of the present invention, or prodrug, or metabolite, or pharmaceutically acceptable salt, can be used in combination as an active ingredient and mixed with a drug carrier to form Pharmaceutical composition. The pharmaceutical carrier can take various forms, depending on the desired mode of administration, for example, oral or injection (including intravenous injection). Therefore, the pharmaceutical composition of the present invention may take the form of a separate unit suitable for oral administration, such as a capsule, cachet or tablet containing a predetermined dose of the active ingredient. Further, the pharmaceutical composition of the present invention may take the form of powder, granule, solution, aqueous suspension, non-aqueous liquid, oil-in-water emulsion, or water-in-oil emulsion. In addition, in addition to the common dosage forms mentioned above, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof can also be administered by a controlled release method and/or a delivery device. The pharmaceutical composition of the present invention can be prepared by any pharmaceutical method. Generally, this method includes the step of associating the active ingredient with the carrier constituting one or more necessary ingredients. In general, the pharmaceutical composition is prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a finely divided solid carrier or a mixture of both. In addition, the product can be easily prepared into the desired appearance.

Therefore, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier and a compound represented by formula (I), or a pharmaceutically acceptable salt thereof. The compound represented by formula (I), or a pharmaceutically acceptable salt thereof, and one or more other compounds having therapeutic activity in combination are also included in the pharmaceutical composition of the present invention.

The drug carrier used in the present invention can be, for example, a solid carrier, a liquid carrier or a gas carrier. Examples of solid carriers include lactose, gypsum powder, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, mannitol, sorbitol, microcrystalline cellulose, inorganic salts , Starch, pregelatinized starch, powdered sugar, dextrin, etc. Examples of liquid carriers include syrup, peanut oil, olive oil and water. Examples of gas carriers include carbon dioxide and nitrogen. When preparing oral pharmaceutical preparations, any convenient pharmaceutical medium can be used. For example, water, ethylene glycol, oils, alcohols, flavor enhancers, preservatives, coloring agents, etc. can be used for oral liquid preparations such as suspensions, elixirs and solutions; and carriers such as starches, sugars, micro Crystal cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, etc. can be used in oral solid preparations such as powders, capsules and tablets. In view of ease of administration, tablets and capsules are preferred for oral preparations. Alternatively, standard aqueous or non-aqueous formulation techniques can be used for tablet coating.

The tablet containing the compound or pharmaceutical composition of the present invention can be prepared by, optionally, mixing with one or more auxiliary components or adjuvants, compression or molding. The active ingredient is in a free-flowing form such as powder or granules, mixed with lubricants, inert diluents, surface active or dispersing agents, and compressed in a suitable machine to obtain compressed tablets. The powdered compound or pharmaceutical composition is soaked with an inert liquid diluent, and then molded in a suitable machine to form a molded tablet. Preferably, each tablet contains about 0.01 mg to 5 g of active ingredient, and each cachet or capsule contains about 0.1 mg to 0.5 g of active ingredient. For example, a dosage form intended for oral administration to humans contains about 0.1 mg to about 0.5 g of active ingredient, compounded with a suitable and convenient metering auxiliary material, which accounts for about 5% to 99.99% of the total pharmaceutical composition . The unit dosage form generally contains about 0.1mg to about 0.5g of effective ingredients, typically 0.1mg, 0.2mg, 0.5mg, 1mg, 2mg, 2.5mg, 5mg, 10mg, 25mg, 50mg, 100mg, 200mg, 300mg, 400mg Or 500mg.

The pharmaceutical composition suitable for parenteral administration provided by the present invention can be prepared as an aqueous solution or suspension by adding active components into water. Suitable surfactants such as sodium lauryl sulfate, polysorbate-80 (Tween-80), polyoxyethylene hydrogenated castor oil, and poloxamer may be included. In glycerin, liquid polyethylene glycol, and their mixture in oil, dispersion systems can also be prepared. Further, a preservative may also be included in the pharmaceutical composition of the present invention to prevent the growth of harmful microorganisms.

The present invention provides pharmaceutical compositions suitable for injection use, including sterile aqueous solutions or dispersion systems. Further, the above-mentioned pharmaceutical composition can be prepared in the form of a sterile powder that can be used for immediate preparation of sterile injection. In any case, the final injection form must be sterile, and for easy injection, it must be easy to flow. In addition, the pharmaceutical composition must be stable during preparation and storage. Therefore, preservation against contamination by microorganisms such as bacteria and fungi is preferable. The carrier can be a solvent or dispersion medium, for example, water, ethanol, polyol (such as glycerol, propylene glycol, liquid polyethylene glycol), vegetable oil, and suitable mixtures thereof.

The pharmaceutical composition provided by the present invention may be in a form suitable for topical administration, for example, aerosol, emulsion, ointment, lotion, dusting powder, or other similar dosage forms. Further, the pharmaceutical composition provided by the present invention can be in a form suitable for use in a transdermal drug delivery device. Using the compound represented by formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, these preparations can be prepared by conventional processing methods. As an example, the preparation of emulsion or ointment is by adding hydrophilic material and water (the total amount of the two is about 5wt% to 50wt% of the compound) to prepare a cream or ointment with the expected consistency.

The pharmaceutical composition provided by the present invention can be made into a form suitable for rectal administration with a solid as a carrier. A unit-dose suppository with the mixture is the best dosage form. Suitable excipients include cocoa butter and other materials commonly used in the art. Suppositories can be conveniently prepared by mixing the pharmaceutical composition with softened or melted auxiliary materials, then cooling and molding.

In addition to the aforementioned carrier components, the above-mentioned pharmaceutical preparations may also include, as appropriate, one or more additional auxiliary components, such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, Lubricants, preservatives (including antioxidants), etc. Further, other adjuvants may also include penetration enhancers that regulate the isotonic pressure between the drug and the blood. The pharmaceutical composition containing the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, can also be prepared in the form of a powder or a concentrated solution.

In order to make the above content clearer and clearer, the present invention will use the following embodiments to further illustrate the technical solution of the present invention. The following examples are only used to illustrate specific implementations of the present invention, so that those skilled in the art can understand the present invention, but are not used to limit the protection scope of the present invention. In the specific embodiments of the present invention, technical means or methods that are not specifically described are conventional technical means or methods in the art.

Unless otherwise specified, all parts and percentages in the present invention are calculated by weight, and all temperatures refer to degrees Celsius.

The following acronyms are used in the examples: ACE-Cl: 1-chloroethyl chloroformate; (BOC) ₂ O: di-tert-butyl dicarbonate; BOP: benzotriazol-1-yloxytri (Dimethylamino)phosphonium hexafluorophosphate; DBU: 1,8-diazabicycloundec-7-ene; DCE: 1,2-dichloroethane; DCM: methylene chloride; DIPEA Or DIEA: N,N-diisopropylethylamine; DMAc: N,N-dimethylacetamide; DMF: N,N-dimethylformamide; DMSO: dimethyl sulfide; EDCI: 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; EtOAc or EA: ethyl acetate; EtOH: ethanol; EtONa: sodium ethoxide; h, hr or hrs: hours; Hex : N-hexane; HOBT: 1-hydroxybenzotriazole; LC-MS or LCMS: liquid chromatography-mass spectrometry; LDA: lithium diisopropylamide; MeCN: acetonitrile; MeOH: methanol; MeONa: sodium methoxide; min or mins: minutes; MsCl: methanesulfonyl chloride; MTBE: methyl tert-butyl ether; MW: microwave; NEt ₃ : triethylamine; NBS: N-bromosuccinimide; NMP: N-methyl -2-pyrrolidone; PdCl ₂ (dppf) ₂ : 1,1'-bisdiphenylphosphine ferrocene palladium dichloride; Pd ₂ (dba) ₃ : tris(dibenzylideneacetone) dipalladium; Pd( OAc) ₂ : Palladium(II) acetate; PE: petroleum ether; PPA: polyphosphoric acid; rt, rt or RT: room temperature; TEA: triethylamine; TFA: trifluoroacetic acid; THF: tetrahydrofuran; Ti(OEt) ₄ : Tetraethyl titanate; TLC: thin layer chromatography; TMEDA: tetramethylethylenediamine; and xantphos: 4,5-bisdiphenylphosphine-9,9-dimethylxanthene.

Preparation of intermediate compound M1:

Step 1: Preparation of compound M1-3

15.00 g of compound M1-1 and 7.08 g of compound M1-2 were dissolved in 150 mL of dioxane, and 198 mg of Pd(OAc) ₂ , 1.70 g of Xantphos and 15.00 g of DIEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 85 ^o C the reaction was stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (50 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 20.00 g of compound M1-3.

Step 2: Preparation of compound M1

M1-3 The compound was dissolved in 20.00g in 200mL THF, -30 ^o C was added dropwise EtONa (35mL, 20% in EtOH) at, RT reaction was stirred for 3hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 200 mL of DCM and stirred for 30 mins, the reaction solution was filtered, and the filter cake was washed with DCM (50 mL×2) to obtain a solid 15g of compound M1.

Preparation of intermediate compound M2:

Step 1: Preparation of compound M2-2

1.00g M2-1 The compound was dissolved in 10mL of DMSO, was added MeONa in MeOH (15mL, 0.5M), and 70 ^o C the reaction 1hr. TLC detected the completion of the reaction. The reaction solution was poured into 30 mL water, extracted with EtOAc (40 mL×3), and the organic phases were combined, washed with 50 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 1.10 g Light yellow oily compound M2-2.

Step 2: Preparation of compound M2-3

1.10g M2-2 The compound was dissolved in 15mL dioxane, is added 491mg compound M1-2,46mg Xantphos, 35mg Pd (OAc) 2 and 1.05g DIEA, mixture was replaced three times with nitrogen, the reaction was heated to 90 ^o C 5hrs. TLC detected that the reaction was complete, the reaction solution was cooled to room temperature, filtered, the filter cake was washed with EtOAc (5 mL×3), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 1.03 g of light yellow solid M2-3.

Step 3: Preparation of compound M2

Dissolve 1.03g of compound M2-3 in 10mL of anhydrous THF, lower the temperature to -30 ^o C, slowly add EtONa EtOH solution (2 mL, 20%) to the above solution, stir and react at -30 ^o C for 30 mins, and slowly rise to The reaction was stirred at room temperature for 2hrs. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was slurried by adding 20 mL of DCM for 30 mins, filtered, the filter cake was washed with DCM (5 mL×3), and the filter cake was vacuum dried to obtain 990 mg of brown solid M2.

Preparation of intermediate compound M3:

Step 1: Preparation of compound M3-2

The compound 1.60g 3.00g Compound M3-1 and M1-2 was dissolved in 30mL dioxane, was added _{243mg Pd 2 (dba) 3,} 384g Xantphos and 3.40g DIPEA, purged with nitrogen three times, the reaction solution was raised to 110 ^o C The reaction was stirred for 3hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (30 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 4.80 g of compound M3-2.

Step 2: Preparation of compound M3

4.80g M3-2 The compound was dissolved in 50mL THF was added dropwise EtONa (7.4mL, 20% in EtOH) at -30 ^o C, RT reaction was stirred for 2hrs. TLC detected the completion of the reaction, concentrated under reduced pressure, added 50 mL DCM and stirred for 30 mins, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), and drained to obtain 3.60 g of compound M3.

Preparation of intermediate compound M4:

Step 1: Preparation of compound M4-3

300 mg of compound M4-1 and 174 mg of compound M4-2 were dissolved in 10 mL of dioxane, and 4 mg of Pd(OAc) ₂ , 34 mg of Xantphos and 300 mg of DIEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 85 ^o C the reaction was stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 266 mg of compound M4-3.

Step 2: Preparation of compound M4

266mg M4-3 The compound was dissolved in 5mL THF in, -30 ^o C was added dropwise under EtONa (0.47mL, 20% in EtOH), RT reaction was stirred for 3hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 15 mL of DCM and stirred for 30 mins, the reaction solution was filtered, and the filter cake was washed with DCM (20 mL×2) to obtain 182 mg of compound M4 as a solid.

Preparation of intermediate compound M5:

Step 1: Preparation of compound M5-3

Under nitrogen, the compound M5-2 10.60g was dissolved in 26mL of ethanol, the reaction solution was cooled 0 ^o C, is added dropwise M5-1 25.00g was added dropwise to the reaction 2hrs RT ice bath was removed, and the reaction was heated overnight at 90 ^o C . TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 4.94 g of a white solid, namely compound M5-3.

Step 2: Preparation of compound M5

M5-3 The compound was dissolved in 100mL DMF, was cooled to 0 ^o C, was added in one 4.98g NBS, then RT reaction 1hr. The reaction was completed by TLC detection. The reaction was quenched by adding 300 mL of water, extracted with EtOAc (300 mL×2), and the organic phases were combined, washed with saturated NaCl (50 mL×4), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography. 5.58g white solid, compound M5.

中間體化合物M6的製備：

Preparation of intermediate compound M6:

Step 1: Preparation of compound M6-3

Under the protection of nitrogen, 25.00g of compound M6-1 was dissolved in 200mL of DMF, the temperature was reduced to 0oC, 22.70g of NaH was added in batches, incubated at 0oC for 1hr, and then 54.96g of compound M6-2 was slowly added dropwise to the reaction solution. After completion, react at 0oC for 1 hr, then heat to 60oC and continue to react for 1 hr. The reaction solution was cooled to 0°C, quenched with 500 mL ice water, extracted with EtOAc (500 mL×3), combined the organic phases, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography Obtained 29.00 g of M6-3 as a brown oil.

Step 2: Preparation of compound M6-5

29.00g of compound M6-3 was dissolved in 50 mL of Ti(OEt)4, 34.99g of compound M6-4 was added, and then heated to 90°C for 12hrs. TCL detected that the reaction was complete. Pour the reaction solution into 500 mL ice water, add 300 mL EtOAc and stir for 1 hr, extract with EtOAc (300 mL×3), combine the organic phases, and wash the organic phase with saturated brine (100 mL×4), anhydrous sodium sulfate Dry and concentrate under reduced pressure to obtain 39.00 g of crude compound M6-5 as a brown oil.

Step 3: Preparation of compound M6-6

Under nitrogen, the compound M6-5 48.00g was dissolved in 500mL dry THF was cooled to -20 ^o C, was slowly added 6.73g NaHB _4, then naturally warmed to RT for 2hrs. After the reaction, the reaction solution was cooled to 0 ^o C, quenched with 300mL of water, (300mL × 3) extracted with EtOAc, the combined organic phases, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, the residue was purified by column Chromatographic purification yielded 25.40 g of compound M6-6 as a brown oil.

Step 4: Preparation of compound M6

10.00 g of compound M6-6 was dissolved in 100 mL of DCM solution, 28.04 g of TFA solution was added dropwise, and then reacted at RT for 1 hr. The reaction solution was cooled to 0 ^o C, with 100mL quenched with saturated aqueous NaHCO _3, EtOAc: THF = 3: Extraction 1 (100mL × 3), combined organic phases were washed with saturated brine and the organic phase was dried over anhydrous sodium sulfate, and reduced pressure It was concentrated to obtain 7.64 g of a brown solid, the crude compound M6, which was directly used in the next reaction.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 7.26-7.21 (m, 4H), 5.80 (d, J = 10.5 Hz, 1H), 4.43 (d, J = 10.5 Hz, 1H), 3.17-3.15 ( m, 2H), 3.08 (d, J = 15.5 Hz, 1H), 2.98-2.88 (m, 2H), 2.69 (d, J = 15.5 Hz, 1H), 2.04-1.99 (m, 1H), 1.80-1.75 (m, 1H), 1.62-1.59 (m, 1H), 1.35(m, 1H), 1.22(s, 9H).

Preparation of intermediate compound M7:

Step 1: Preparation of compound M7-3

10.00 g of compound M7-1 and 19.50 g of compound M7-2 were dissolved in 100 mL of MeCN, and 26.20 g of K ₂ CO _{3 was added} . The reaction was raised to 90 ^o C the reaction was stirred for 3hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with EtOAc (50 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 5.90 g of compound M7-3.

Step 2: Preparation of compound M7-4

1.50g M7-3 The compound was dissolved in 15mL of toluene was added dropwise 1.1mL PBr _3, the reaction liquid was raised to 105 ^o C the reaction was stirred for 12hrs. TLC detects the completion of the reaction, concentrated under reduced pressure, added 15 mL of water, adjusted to pH=9 with NaOH solution, extracted with EtOAc (30 mL×3), combined the organic phases, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 1.50 g of compound M7-4 .

Step 3: Preparation of compound M7-6

450mg of M7-5 was dissolved in 6mL DMF, the lower 0 ^o C was added 271mg NaH, under nitrogen, 60 ^o C After stirring for 1hr the reaction was added 1.20g Compound M4-4,60 ^o C the reaction was stirred for 1hr. TLC detected the completion of the reaction. The reaction was quenched by adding 30 mL of water, extracted with EtOAc (25 mL×2) and water (30 mL×3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 160 mg Compound M7-6.

Step 4: Preparation of compound M7-7

At 0 ^o C M7-6 The compound 160mg was dissolved in 2mL DCE and added dropwise to 155mg ACE-Cl, RT reaction was stirred for 2hrs. TLC the reaction was complete, concentrated under reduced pressure, 4 mL MeOH was added, the reaction was raised to 80 ^o C the reaction was stirred for 3hrs. The reaction was completed by TLC detection, concentrated under reduced pressure, 4 mL DCM, 242 mg (Boc) ₂ O and 239 mg DIEA were added, and the reaction was stirred at RT for 12 hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 25 mg of compound M7-7.

Step 5: Preparation of compound M7

The procedure for preparing compound M7 from compound M7-7 is similar to the procedure from compound M6-3 to compound M6.

Preparation of intermediate compound M8:

Step 1: Preparation of compound M8-3

4.00g M8-1 The compound was dissolved in 50mL anhydrous THF, purged with nitrogen three times, cooled to -78 ^o C, was slowly added dropwise a solution of LDA in THF (11.70mL, 2.0M), the reaction for 1 hr and then -78 ^o C, the compound M8-2 in THF (10mL) was slowly added dropwise to the reaction solution, -78 ^o C the reaction 30mins, the reaction was slowly warmed to room temperature 2hrs. TLC detected the completion of the reaction, quenched the reaction with 30mL saturated NH ₄ Cl solution, added 50mL water, extracted with EtOAc (60mL×3), combined the organic phases, washed with 50mL saturated NaCl, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and left the residue Purified by column chromatography to obtain 4.30 g of colorless oil M8-3.

Step 2: Preparation of compound M8-4

4.30g M8-3 The compound was dissolved in THF / MeOH (40mL / 40mL) was added aqueous NaOH (20mL, 2.4N), the reaction was heated to 80 ^o C 18hrs. The reaction was completed by TLC detection, the reaction solution was cooled to room temperature, concentrated under reduced pressure and evaporated to remove the organic solvent, the residue was adjusted to pH 3-4 with concentrated hydrochloric acid, filtered, the filter cake was washed with water (10 mL×3), and the filter cake was vacuumed After drying, 3.40 g of compound M8-4 was obtained as a white solid.

Step 3: Preparation of compound M8-5

3.40g M8-4 The compound was dissolved in 40mL PPA, the reaction temperature was raised to 120 ^o C 2hrs. TLC detected that the reaction was complete, and the reaction solution was slowly added dropwise to 200 mL of crushed ice, adjusted to pH 9-10 with 2.4N NaOH aqueous solution, 4.40 g (B _OC ) ₂ O was added, and the reaction was stirred at RT for 18 hrs. TLC detected that the reaction was complete, extracted with EtOAc (100 mL×3), combined the organic phases, washed with 100 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 2.40 g of white solid M8-5.

Step 4: Preparation of compound M8

The procedure for preparing compound M8 from compound M8-5 is similar to the procedure from compound M6-3 to compound M6.

Preparation of intermediates M9-a and M9-b:

Step 1: Preparation of Compound 10.00g Compound M9-1 M9-2 will be dissolved in 100mL MeOH was added 2.0mL of concentrated sulfuric acid, 70 ^o C the reaction was heated 3 hrs. After the reaction is complete, spin off the solvent, add 20 mL of water, adjust to pH=9 with saturated aqueous Na ₂ CO ₃ , extract with EtOAc (100 mL×3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and pass the residue through a column Chromatographic purification yielded 9.23 g of white solid, compound M9-2.

Step 2: Preparation of compound M9-3

9.23g M9-2 The compound was dissolved in 150mL MeOH in an ice bath and cooled to 0 ^o C, was added 6.97g NaBH _4, the reaction was naturally warmed to RT 5hrs. After the reaction is complete, add 20 mL of saturated NH ₄ Cl solution, spin dry the solvent, add EtOAc (100 mL×3) for extraction, combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain 6.40 g of colorless liquid , Namely compound M9-3.

Step 3: Preparation of compound M9-4

Under nitrogen, the compound M9-3 3.00g dissolved in 50mL of dichloromethane, the reaction solution was cooled to -15 ^o C, was added 2.81mL NEt _3, and then 1.04mL of a solution of MsCl dropwise. After the addition was complete, warmed to Reaction at 0 ^o C for 1 hr. After the completion of the reaction, a water layer was added, the organic phase was washed with 20 mL of saturated brine, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 4.05 g of white solid compound M9-4.

Step 4: Preparation of compound M9-6

Under nitrogen, the compound M9-5 3.39g dissolved in 20mL dry THF was cooled to -50 ^o C, was added dropwise 1.71g LDA solution was added dropwise to the -50 ° C reaction incubated 1hr. A solution of 3.00 g of compound M9-4 in anhydrous THF (10 mL) was added dropwise, after the dropping, the temperature was raised to RT and reacted for 1 hr. After the reaction was completed, 50 mL of brine was added, extracted with EtOAc (50 mL×3), the organic phases were combined, the organic phase was dried with sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 4.61 g of white solid compound M9-6.

Step 5: Preparation of compounds M9-7a and M9-7b

4.61 g of compound M9-6 was dissolved in 8 mL of water and 40 mL of MeOH, and 2.07 g of NaOH was added. Warm to 65 ^o C and stir overnight. After the reaction is complete, add 30 mL of water, spin dry the solvent methanol, adjust the concentrate to pH=6 with 2N hydrochloric acid, extract with EtOAc (50 mL×3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and pass the residue through the column Analytical purification yielded 3.98 g of white solids, namely compounds M9-7a and M9-7b.

Step 6: Preparation of compounds M9-8a and M9-8b

Under the protection of nitrogen, dissolve 3.98g of the mixture of M9-7a and M9-7b in 20mL of anhydrous THF, cool the reaction solution to -15 ^o C, add NaH (60%, 0.42g) in batches, and then keep the reaction at -15 ^o C 1hr, then cooled to -60 ^o C, n-butyllithium (1.6M, 7.8mL), the reaction incubated 1hr. After the reaction is complete, add 50 mL of water, extract with EtOAc (50 mL×3), combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain 2.36 g of pale yellow solid, which is a mixture of M9-8a and M9 -8b.

Step 7: Preparation of compounds M9-a and M9-b

The procedure for preparing M9-a and M9-b from M9-8a and M9-8b is similar to the procedure from compound M6-3 to compound M6.

Preparation of intermediate compound M10:

Step 1: Preparation of compound M10-3

Under nitrogen atmosphere, 2.83g M10-2 compound was dissolved in 50mL of anhydrous THF, cooled to -78 ^o C, was added dropwise LDA (2M, 6mL) in THF / Hex solution. Incubated for 1 hr at -78 ^o C, and then 1.69g of compound M10-1 THF (3mL) was slowly added dropwise to the reaction solution was added dropwise to the reaction at -78 ^o C 1hr. The reaction solution was quenched with 50 mL saturated brine, extracted with EtOAc (30 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 1.44 g of a pale yellow oil compound M10- 3.

Step 2: Preparation of compound M10-4

Under nitrogen, the compound M10-3 900mg was dissolved in 50mL anhydrous THF was cooled to -78 ^o C. A THF/Hex solution of LDA (2M, 3 mL) was added dropwise. Incubate the reaction at -78 ^o C for 1 hr. The reaction solution was quenched with 50 mL saturated brine, extracted with EtOAc (30 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 300 mg of light yellow solid compound M10-4.

Step 3: Preparation of compound M10

The procedure for preparing compound M10 from compound M10-4 is similar to the procedure from compound M6-3 to compound M6.

Through different reaction starting materials and suitable reagents, for example, the starting material for the synthesis of M12 is 6-methoxy-1-indanone, and the intermediate compounds in Table 1 are prepared by a method similar to the aforementioned intermediate 6-10 M11-M15.

M12

M13

M14

M15

      Table 1 Intermediate Number Chemical structure Intermediate Number Chemical structure M11

M12

M13

M14

M15

Example 2: Compound (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3- Preparation of dichlorophenyl)thio)-5-methylpyrazin-2-yl)methanol

Step 1: Preparation of compound 2-2

500 mg of compound 2-1 and 729 mg of compound M5 were dissolved in 15 mL of dioxane, and 127 mg of Pd ₂ (dba) ₃ , 161 mg of Xantphos and 1 g of DIEA were added. Nitrogen thrice, with stirring under nitrogen for 12hrs at 80 ^o C. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with EtOAc (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 350 mg of compound 2-2.

Step 2: Preparation of compound 2-3

228mg The compound 2-2 was dissolved in 3mL POCl ₃ was added 38mg compound N, N- dimethylanilinium Amides, the reaction was raised to 110 ^o C with stirring 1.5hrs. TLC detects the completion of the reaction, concentrates under reduced pressure, adds NaHCO ₃ aqueous solution to adjust the pH to 7-8 in an ice bath, and extracts with EtOAc/THF=1/1 (30mL×2). Combine the organic phases, dry with anhydrous sodium sulfate, and reduce pressure. Concentrated to obtain 195 mg of compound 2-3.

Step 3: Preparation of compound 2-4

195 mg of compound 2-3 and 236 mg of compound M6 were dissolved in 3 mL of NMP, and 268 mg of DIEA was added. The reaction was raised to 80 ^o C was stirred for 3hrs. TLC detected that the reaction was complete, extracted with EtOAc/THF=1/1 (25 mL×2), combined the organic phases, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 165 mg of compound 2-4.

Step 4: Preparation of compound 2-5

At 0 ^o C to 165mg Compound 2-4 was dissolved in 2.5mL THF was added portionwise 24mg LiAlH _4, stirred for 10mins. The reaction was complete by TLC, at 0 ^o C were added 50 L of water, NaOH (15%, 50μL) , 150μL water, and 1g of anhydrous sodium sulfate, stirred for 10mins. After filtration, the filter cake was washed with THF/DCM=1/1 (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 94 mg of compound 2-5.

Step 5: Preparation of compound 2

94 mg of compound 2-5 was dissolved in 1.5 mL of dioxane, 0.4 mL of 2N HCl in methanol was added, and reacted at RT for 1 hr. TLC detects the completion of the reaction, concentrates under reduced pressure, adds NaHCO ₃ aqueous solution to adjust the pH to 7-8, extracts with DCM/MeOH=10/1 (15mL×3), combines the organic phases, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue After purification by column chromatography, 47.9 mg of compound 2 was obtained.

[M+H ⁺ ]=501.13.

¹ H NMR (500 MHz, DMSO- d ₆ ): δ 7.48 (d, J = 8.5 Hz, 1H), 7.32-7.30(m, 1H), 7.25 (t, J = 8.5 Hz, 1H), 7.18-7.13 (m, 3H), 6.77 (d, J = 8.5 Hz, 1H), 5.35 (t, J = 5.5 Hz, 1H), 4.46(d, J = 5.5 Hz, 1H), 3.90-3.83(m, 3H) , 3.18-3.09(m, 2H), 3.04(d, J = 16.0 Hz, 1H), 2.62(d, J = 16.0Hz, 1H), 2.41(s, 3H), 1.92-1.86(m, 1H), 1.80-1.75(m, 1H), 1.55-1.53(m, 1H), 1.16-1.13(m, 1H).

Example 3: Compound (S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazine-2-carbonitrile preparation

Step 1: Preparation of compound 3-2

At RT, 174 mg of compound 3-1 and 306 mg of compound M6 were dissolved in 3 mL of anhydrous THF, and 0.28 mL of NEt _{3 was} added. After nitrogen replacement three times, RT reaction for 1 hr. After the reaction was completed, 10 mL of water and 10 mL of EtOAc were added for extraction three times, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 142 mg of yellow oily compound 3-2.

Step 2: Preparation of compound 3-3

The compound 142mg and 55mg KI M1 3-2,62.3mg compound dissolved in dioxane in a microwave reactor 120 ^o C 1hr. LC-MS detected that the reaction was complete, added 10 mL of water and 10 mL of EtOAc for extraction three times, combined the organic layers, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 121 mg of yellow solid compound 3-3.

Step 3: Preparation of compound 3

60 mg of compound 3-3 was dissolved in 2.0 mL of 1,4-dioxane, 0.3 mL of 2M hydrochloric acid in methanol was added dropwise under RT conditions, and the reaction was carried out for 30 mins under RT conditions. After the reaction is complete, spin off the solvent, add 1 mL of water, adjust the pH to 8 with saturated aqueous NaHCO ₃ solution, add 10 mL of water after the solid is precipitated, and extract with EtOAc (10 mL×3). Combine the organic phases, dry with anhydrous sodium sulfate, and reduce pressure. After concentration, the residue was purified by column chromatography to obtain 30.2 mg of compound 3 as a yellow solid.

[M+H ⁺ ]=464.19.

¹ H NMR (500 MHz, CDCl3): δ 8.34 (s, 1H), 7.79 (d, J = 5.5 Hz, 1H), 7.35-7.33 (m, 1H), 7.25 (m, 3H), 6.17 (d, J = 5.5 Hz, 1H), 4.56-4.50 (m, 2H), 4.03 (s, 1H), 3.51-3.42 (m, 2H), 3.12 (d, J = 15.5 Hz, 1H), 2.77 (d, J = 15.5 Hz, 1H), 2.01-1.94 (m, 1H), 1.91- 1.83 (m, 1H), 1.47-1.44(m, 1H), 1.26-1.24(m, 1H).

Example 4: Compound ( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-di Preparation of chlorophenyl)thio)pyrazine-2-formamide

Step 1: Preparation of compound 4-2

200mg and Compound 4-1 to Compound 2-1 of 104.86mg DMA (3mL) was added and stirred at 14.96mg KI ^{_{186.77mg K 2 CO 3, 120 o}} C 2hrs. The reaction solution was diluted with 10 mL of water, extracted with EtOAc (10 mL×3), and the organic phases were combined. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated. The crude product was separated and purified by Pre-TLC (PE:EtOAc=3:1) 233 mg of compound 4-2 was obtained as a yellow solid, and the yield was 88.18%.

[M+H ⁺ ]:586.23.

Step 2: Preparation of compound 4-3

Was added dropwise to a mixed solution of NaOH (1.56 mL) and water (1.56 mL) 200mg of compound 4-2 in dioxane (2.5M, 132.97μL), stirred at 100 ^o C 12hrs. The reaction solution was adjusted to pH 6-7 with 1N HCl, and extracted with EtOAc (10mL×3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated. The crude product was subjected to Pre-TLC (DCM:MeOH=20 1) Separation and purification to obtain 93 mg of compound 4-3 as a yellow solid with a yield of 69.41%.

[M+H ⁺ ]:605.29.

Step 3: Preparation of compound 4

At 0 ^o C, methanol was added dropwise a solution of hydrochloric acid (2M, 0.5mL) to compound 95mg dioxane (3mL) 4-3 was stirred for 1hr at 25 ^o C. The reaction solution was quenched by adjusting the pH to 8-9 with saturated aqueous NaHCO ₃ solution, extracted with EtOAc:THF=5:1 (10 mL×3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate and concentrated The crude product was separated and purified by Pre-TLC (DCM:MeOH=20:1) and then separated and purified by Pre-HPLC (basic) to obtain 36 mg of compound 4 as a yellow solid with a yield of 45.32%.

[M+H ⁺ ]=500.11

¹ H NMR (500 MHz, CDCl ₃ ): δ 8.20 (s, 1H), 7.35-7.33 (dd, J = 8.0, 1.5 Hz, 1H), 7.32 (s, 1H), 7.25-7.19 (m, 4H) , 7.11 (t, J = 8.0 Hz, 1H), 6.99 (dd, J = 8.0, 1.5 Hz, 1H), 5.40 (s, 1H), 4.06-3.93 (m, 3H), 3.39-3.29 (m, 2H) ), 3.11 (d, J = 16.0 Hz, 1H), 2.74 (d, J = 16.0 Hz, 1H), 1.93-1.87 (m, 1H), 1.83-1.78 (m, 1H), 1.63-1.59 (m, 1H), 1.38-1.35 (m, 1H).

Example 5: Compound ( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazine-2-carboxamide preparation

Step 1: Preparation of compound 5-1

121 mg of compound 3-3 was dissolved in 1.2 mL of 1,4-dioxane and 1.2 mL of water, and 0.12 mL of 2.5 M NaOH aqueous solution was added. The reaction was heated to 100 ^o C the reaction 6hrs. TLC detected the completion of the reaction, added 10 mL of water and 10 mL of EtOAc for extraction three times, combined the organic layers, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 32 mg of yellow solid compound 5-1.

Step 2: Preparation of compound 5

Under RT conditions, 32 mg of compound 5-1 was dissolved in 1.0 mL of 1,4-dioxane, 0.2 mL of 2M HCl in methanol was added dropwise, and reacted for 30 mins at RT. After the reaction was complete, the solvent was spin-dried, 1 mL of water was added, and the pH was adjusted to 8 with saturated aqueous NaHCO ₃ solution. After the solid precipitated out, 10 mL of water was added, extracted with EtOAc (10 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 16 mg of compound 5 as a yellow solid.

[M+H ⁺ ]=482.19.

¹ H NMR (500 MHz, DMSO- d ₆ ): δ 8.34 (s, 1H), 8.04 (s, 1H), 7.69-7.67 (m, 2H), 7.32 (d, J = 6.5 Hz, 1H), 7.19 -7.17 (m, 3H), 6.36 (s, 2H), 5.89 (d, J = 5.0 Hz, 1H), 4.01 (t, J = 11.5 Hz, 2H), 3.87 (s, 1H), 3.28-3.23 ( m, 2H), 3.08 (d, J = 15.5 Hz, 1H), 2.66 (d, J = 15.5 Hz, 1H), 1.83-1.70 (m, 2H), 1.53-1.51 (m, 1H),1.16-1.14 (m, 1H).

Using different reaction starting materials and appropriate reagents, the compounds 7-18, 58 and 59 in Table 2 were prepared by a method similar to that of the foregoing Example 5.

(S )-3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((3-氯-2-甲氧基吡啶-4-基)硫基)吡嗪-2-甲醯胺 497.18 8

(S )-3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-甲醯胺 501.20 9

(S )-3-(1-氨基-6-甲氧基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-甲醯胺 531.16 10

(S )-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-溴-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-甲醯胺 562.07 11

(S )-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-甲氧基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-甲醯胺 512.17 12

(S )-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-甲氧基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-甲醯胺 512.18 13

(S )-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-5,6-二甲基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-甲醯胺 542.12 14

(S )-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-(三氟甲基)-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-甲醯胺 550.16 15

(S )-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氯-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-甲醯胺 516.14 16

(S )-3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((6-氨基-3-氯吡啶-2-基)硫基)吡嗪-2-甲醯胺 482.18 17

(S )-3-(5-氨基-2-甲氧基-5,7-二氫螺[環戊二烯[b ]吡啶-6,4'-呱啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲醯胺 513.18 18

(S )-3-(4-氨基-2-氯-4,6-二氫螺[環戊二烯并[d ]噻唑-5,4'-呱啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲醯胺 523.09 58

(S)-3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-氨基-3-氟吡啶-4-基)硫基)吡嗪-2-甲醯胺 466.17 59

(S)-3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((3-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-甲醯胺 562.19 Table 2 Compound number Chemical structure Chemical Name LC-MS [M+H ⁺ ] 7

( S )-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-chloro-2-methoxypyridine -4-yl)thio)pyrazine-2-carboxamide 497.18 8

( S )-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-(trifluoromethyl)pyridine- 3-yl)thio)pyrazine-2-carboxamide 501.20 9

( S )-3-(1-Amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(三(Fluoromethyl)pyridin-3-yl)thio)pyrazine-2-carboxamide 531.16 10

( S )-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-bromo-1,3-dihydrospiro(indene-2,4'-Pyridine]-1'-yl)pyrazine-2-carboxamide 562.07 11

( S )-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-methoxy-1,3-dihydrospiro(indene-2, 4'-piperidine)-1'-yl)pyrazine-2-methamide 512.17 12

( S )-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-methoxy-1,3-dihydrospiro(indene-2, 4'-piperidine)-1'-yl)pyrazine-2-methamide 512.18 13

( S )-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro(indene- 2,4'-piperidine)-1'-yl)pyrazine-2-methamide 542.12 14

( S )-6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-(trifluoromethyl)-1,3-dihydrospiro(indene -2,4'-pyridine)-1'-yl)pyrazine-2-carboxamide 550.16 15

( S )-6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2,4'-Pyridine]-1'-yl)pyrazine-2-carboxamide 516.14 16

( S )-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((6-amino-3-chloropyridine-2 -Yl)thio)pyrazine-2-carboxamide 482.18 17

( S )-3-(5-Amino-2-methoxy-5,7-dihydrospiro[cyclopentadiene[ b ]pyridine-6,4'-piperidine]-1'-yl)-6 -((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide 513.18 18

( S )-3-(4-Amino-2-chloro-4,6-dihydrospiro[cyclopenta[ d ]thiazole-5,4'-piperidine]-1'-yl)-6- ((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide 523.09 58

(S)-3-(1-Amino-1,3-dihydrospiro[inden-2,4'-piperidine]-1'-yl)-6-((2-amino-3-fluoropyridine-4 -Yl)thio)pyrazine-2-carboxamide 466.17 59

(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-chloro-2-((1- Methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)sulfanyl)pyrazine-2-carboxamide 562.19

The NMR data of compounds 7, 8, 9, 10, 11, 12, 14 and 59 are as follows:

¹ H NMR (500 MHz, DMSO- d ₆ ): δ 8.39 (s, 1H), 8.06 (s, 1H), 7.92 (d, J = 5.5 Hz, 1H), 7.69 (s, 1H), 7.31-7.30 (m, 1H), 7.19-7.16 (m, 3H), 6.39 (d, J = 5.5 Hz, 1H), 4.07-3.99 (m, 2H), 3.93 (s, 3H), 3.85 (s, 1H), 3.29-3.24 (m, 2H), 3.07 (d, J = 15.4 Hz, 1H), 2.64 (d, J = 15.4 Hz, 1H), 1.83-1.79 (m, 1H), 1.74-1.70 (m, 1H) , 1.55-1.52 (m, 1H), 1.15-1.13 (m, 1H). (Compound 7)

¹ H NMR (500 MHz, DMSO- d ₆ ): δ 8.55 (d, J = 4.5 Hz, 1H), 8.33 (s, 1H), 8.01 (s, 1H), 7.71-7.67(m, 2H) ,7.63 -7.62(m, 1H) ,7.30( d, J = 6.5 Hz, 1H), 7.17-7.14(m, 3H), 4.01-3.97(m, 2H), 3.84(s, 1H), 3.28-3.23(m , 2H), 3.05(d, J = 15.0 Hz, 1H), 2.63(d, J = 14.5 Hz, 1H), 1.80-1.77(m, 1H), 1.70-1.67(m, 1H), 1.52-1.50( m, 1H), 1.13-1.10(m, 1H). (Compound 8)

¹ H NMR (500 MHz, CDCl ₃ ) :δ 8.55 (dd, J = 4.5, 1.0 Hz, 1H), 8.22 (s, 1H), 7.67 (d, J = 7.5 Hz,1H), 7.39 (dd, J = 8.2, 4.5 Hz, 1H), 7.12 (br, 1H), 7.10 (d, J = 8.3 Hz, 1H), 6.89 (d, J = 2.5 Hz, 1H), 6.76 (dd, J = 8.2, 2.5 Hz , 1H), 5.36 (br, 1H), 4.03-4.98 (m, 2H), 3.95 (s, 1H), 3.81 (s, 3H), 3.38-3.27 (m, 2H), 3.03 (d, J = 15.0 Hz, 1H), 2.67 (d, J = 15.0 Hz, 1H), 1.92-1.86 (m, 1H), 1.80-1.74 (m, 1H), 1.62-1.59 (m, 1H), 1.36-1.33 (m, 1H). (Compound 9)

1H NMR (500 MHz, DMSO- d ₆ ): δ 8.36 (s, 1H), 8.04 (s, 1H), 7.69 (m, 2H), 7.38 -7.13 (m, 3H), 6.38 (s, 2H), 5.89 (d, J = 5.0 Hz, 1H), 4.03-3.97 (m, 2H), 3.94 (s, 1H), 3.32-3.24 (m, 2H), 3.04 (d, J = 17.0 Hz, 1H), 2.65 (d, J = 17.0 Hz, 1H), 1.82-1.72 (m, 2H), 1.57-1.54 (m, 1H), 1.16-1.14 (m, 1H). (Compound 10)

1H NMR (500 MHz, DMSO- d ₆ ): δ 8.35 (s, 1H), 8.05 (s, 1H), 7.69 (m, 2H), 7.19 (t, J = 7.5 Hz, 1H), 6.95 (d, J = 7.5 Hz, 1H), 6.82 (d, J = 7.5 Hz, 1H), 6.38 (s, 2H), 5.88 (d, J = 5.0 Hz, 1H), 4.05-3.97 (m, 2H), 3.91 ( s, 1H), 3.77 (s, 3H), 3.27-3.24 (m, 2H), 2.96(d, J = 16.0 Hz, 1H), 2.58 (d, J = 16.0 Hz, 1H), 1.79-1.75 (m , 2H), 1.49-1.51 (m, 1H), 1.23-1.21 (m, 1H). (Compound 11)

¹ H NMR (500 MHz, CDCl ₃ ) :δ 8.28 (s, 1H), 7.75 (d, J = 5.0 Hz, 1H), 7.12(d, J = 8.5 Hz, 1H), 6.89(d, J = 2.0 Hz, 1H), 6.77(dd, J = 8.5 Hz, J = 2.0 Hz, 1H), 6.07(d, J = 5.5 Hz, 1H), 4.90 (s, 2H), 4.08-4.03(m, 2H), 3.96 (s, 1H), 3.81 (s, 3H), 3.41-3.31(m, 2H), 3.04(d, J = 15.0 Hz, 1H), 2.68(d, J = 15.0 Hz, 1H), 1.94-1.88 (m, 1H), 1.82-1.77(m, 1H), 1.64-1.62(m, 1H), 1.38-1.35(m, 1H). (Compound 12)

¹ H NMR (500 MHz, CDCl ₃ ): δ 8.29 (s, 1H), 7.74 (d, J = 5.4 Hz, 1H), 7.60 (s, 1H), 7.49 (d, J = 7.8 Hz, 1H), 7.32 (d, J = 7.8 Hz, 1H), 6.07 (d, J = 5.4 Hz, 1H), 4.16-4.04 (m, 2H), 4.03 (s, 1H), 3.43-3.29 (m, 2H), 3.18 (d, J = 16.0 Hz, 1H), 2.79 (d, J = 16.0 Hz, 1H), 1.98-1.92 (m, 1H), 1.85-1.80 (m, 1H), 1.65-1.62 (m, 1H), 1.35-1.31(m, 1H). (Compound 14)

[M+H ⁺ ]=562.20.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.51 (s, 1H), 8.38 (s, 1H), 8.03 (s, 1H), 7.78 (d, J = 5.4 Hz, 1H), 7.71-7.66 ( m, 1H), 7.36 (d, J = 1.9 Hz, 1H), 7.33-7.27 (m, 1H), 7.21-7.13 (m, 3H), 6.15 (d, J = 5.4 Hz, 1H), 6.09 (d , J = 1.9 Hz, 1H), 4.07-3.99 (m, 2H), 3.84 (s, 1H), 3.56 (s, 3H), 3.30 -3.22 (m, 2H), 3.07 (d, J = 15.6 Hz, 1H), 2.64 (d, J = 15.6 Hz, 1H), 1.85-1.78 (m, 1H), 1.76-1.68 (m, 1H), 1.55-1.50(m, 1H), 1.17-1.11(m, 1H) . (Compound 59)

Preparation of intermediate compound M15-3:

Step 1: Preparation of compound M15-2

530 mg of compound M15-1 and 190 mg of compound M1-2 were dissolved in 20 mL of dioxane, and 73 mg of Pd ₂ (dba) ₃ , 92 mg of Xantphos and 410 mg of DIEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 85 ^o C the reaction was stirred for 12hrs. The reaction was completed by LCMS, the reaction solution was filtered, 75mL saturated brine was added to the filtrate, extracted with EtOAc (20mL×4), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography. 510 mg of compound M15-2.

Step 2: Preparation of compound M15-3

250 mg of compound M15-2 was dissolved in 3 mL of THF, EtONa (1.4 mL, 20% EtOH solution) was added dropwise at RT, and the reaction was stirred at RT for 45 mins. The reaction was completed by LCMS detection, concentrated under reduced pressure, 5 mL of DCM was added and ultrasonicated for 5 mins, the supernatant was decanted after standing, and 5 mL of MTBE was added for 5 minutes of sonication. After standing, the supernatant was decanted and dried to obtain 116 mg of compound M15-3 as a solid.

Preparation of intermediate compound M16-5:

Step 1: Preparation of compound M16-3

The 1.90g NaH (60% content) was added 25mL THF under nitrogen the reaction was raised to 60 ^o C, and a solution of 5.00g 3.46g Compound M16-1 M16-2 compound of THF (40mL) solution, the reaction was stirred for 2.5hrs . TLC detected the completion of the reaction. The reaction solution was poured into 20 mL of water, extracted with DCM (40 mL×2), and the organic phases were combined, washed with 30 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 4.45 g yellow solid compound M16-3.

Step 2: Preparation of compound M16-4

Dissolve 4.45g compound M16-3 and 2.05g compound M1-2 in 50mL dioxane, add 283mg Pd ₂ (dba) ₃ , 448mg Xantphos and 4.00g DIPEA, replace with nitrogen three times, and raise the reaction solution to 85 ^o C The reaction was stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (20 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 5.38 g of compound M16-4.

Step 3: Preparation of compound M16-5

The M16-4 compound was dissolved in 5.38g in 50mL THF, -30 ^o C was added dropwise under EtONa (9mL, 20% in EtOH), RT reaction was stirred for 2hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 30 mL DCM and stirred for 30 mins, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), and drained to obtain 4.29 g of compound M16-5.

Preparation of intermediate compound M17-3:

Step 1: Preparation of compound M17-1

The 700 mg Compound M17-SM and 2.5 mL NH (CH _{3) 2} was dissolved in an aqueous solution of 2.5 mL dioxane, purged with nitrogen three times, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 2 hr. The reaction was completed by TLC detection, 10 mL water and 10 mL ethyl acetate were added for extraction, the organic phase was separated, washed with 10 mL saturated sodium chloride solution, and dried over anhydrous sodium sulfate. The solvent was spun off to obtain 708 mg of compound M17-1.

Step 2: Preparation of compound M17-2

708 mg of compound M17-1 and 320 mg of compound M1-2 were dissolved in 10 mL dioxane, and 243 mg Pd ₂ (dba) ₃ , 308 mg Xantphos and 688 mg DIEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 5 hr. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 584 mg of compound M17-2.

Step 3: Preparation of compound M17-3

Dissolve 584 mg of compound M17-2 in 6 mL THF, add dropwise an ethanol solution (4 mL) of EtONa (185 mg) and CH ₃ ONa (183 mg) at room temperature, and react with stirring at RT for 2 hr. TLC detects that the reaction is complete, concentrates under reduced pressure to obtain a solid, adds 10 mL of methyl tert-butyl ether and stirs for 5 min, removes the reaction liquid, repeats three times, and drains to obtain a solid 500 mg of crude compound M17-3, which can be used directly for the next One step response.

Preparation of intermediate compound M18-3:

Step 1: Preparation of compound M18-1

The 700 mg Compound M17-SM and 2.5 mL methylamine solution dissolved in 2.5 mL dioxane, purged with nitrogen three times, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 2 hr. The reaction was completed by TLC detection, 10 mL water and 10 mL ethyl acetate were added for extraction, the organic phase was separated, washed with 10 mL saturated sodium chloride solution, and dried over anhydrous sodium sulfate. The solvent was spun off to obtain 728 mg of compound M18-1.

Step 2: Preparation of compound M18-2

728 mg of compound M18-1 and 347 mg of compound M1-2 were dissolved in 7 mL of dioxane, and 264 mg Pd ₂ (dba) ₃ , 334 mg Xantphos and 747 mg DIEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 5 hr. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 646 mg of compound M18-2.

Step 3: Preparation of compound M18-3

Dissolve 646 mg of compound M18-2 in 7 mL of THF, add dropwise EtONa (180 mg) and CH ₃ ONa (243 mg) in ethanol (3 mL) at room temperature, and stir for 2 hr at RT. TLC detects that the reaction is complete, and concentrates under reduced pressure to obtain a solid. Add 10 mL of methyl tert-butyl ether and stir for 5 min. Remove the reaction liquid. Repeat three times and drain to obtain a solid 500 mg of crude compound M18-3, which can be used directly for the next One step response.

Preparation of intermediate compound M19-2:

Step 1: Preparation of compound M19-1

Dissolve 1 g of compound M19-SM and 578 mg of compound M1-2 in 10 mL of dioxane, and add 220 mg of Pd ₂ (dba) ₃ , 278 mg of Xantphos and 1.24 g of DIEA. Nitrogen thrice, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 5 hr. TLC detected the completion of the reaction, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 885 mg of compound M19-1.

Step 2: Preparation of compound M19-2

885 mg of compound M19-1 was dissolved in 8 mL of THF, and EtONa (268 mg) and CH ₃ ONa (251 mg) in ethanol (4 mL) were added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detects that the reaction is complete, and concentrates under reduced pressure to obtain a solid. Add 10 mL of methyl tert-butyl ether and stir for 5 min. Remove the reaction liquid. Repeat three times and drain to obtain a solid 700 mg of crude compound M19-2, which can be used directly for the next step. One step response.

Preparation of intermediate compound M20-3:

Step 1: Preparation of compound M20-1

1 g of the compound M20-SM and ₃ mL NH (CH 3) ₂ was dissolved in 3 mL aqueous dioxane, purged with nitrogen three times, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 2 hr. The reaction was completed by TLC detection, 10 mL water and 10 mL ethyl acetate were added for extraction, the organic phase was separated, washed with 10 mL saturated sodium chloride solution, and dried over anhydrous sodium sulfate. The solvent was spun off to obtain 1.03 g of compound M20-1.

Step 2: Preparation of compound M20-2

1.03 g of compound M20-1 and 438 mg of compound M1-2 were dissolved in 10 mL of dioxane, and 334 mg of Pd ₂ (dba) ₃ , 422 mg of Xantphos and 942 mg of DIEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 4 hr. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 968 mg of compound M20-2.

Step 3: Preparation of compound M20-3

968 mg of compound M20-2 was dissolved in 10 mL of THF, and EtONa (120 mg) and CH ₃ ONa (228 mg) in ethanol (3 mL) were added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detects that the reaction is complete, and concentrates under reduced pressure to obtain a solid. Add 10 mL of methyl tert-butyl ether and stir for 5 min. Remove the reaction liquid. Repeat three times and drain to obtain a solid 900 mg of crude compound M20-3, which can be used directly for the next step. One step response.

Preparation of intermediate compound M21-3:

Step 1: Preparation of compound M21-1

4.0 g of compound M17-SM was dissolved in 20 mL of 25% ammonia solution and 20 mL of dioxane, and the tube was sealed at 70 ^o C and the reaction was stirred for 48 hr. TLC detects that the reaction is complete, the reaction solution is added with water (20ml), extracted with EA (50ml×2), the organic phase is washed with saturated brine (10ml×2), dried with anhydrous sodium sulfate, spin-dried, the solid is slurried with n-hexane and filtered to obtain 3.67g , Namely compound M21-1.

Step 2: Preparation of compound M21-2

Dissolve 2.92 g of compound M21-1 and 1.47 g of compound M1-2 in 45 mL of dioxane, and add 354 mg of Pd ₂ (dba) ₃ , 225 mg of Xantphos and 3.17 g of DIEA. Nitrogen thrice, under nitrogen, the reaction was raised to 80 ^o C and stirred overnight. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 2.68 g of compound M21-2.

Step 3: Preparation of compound M21-3

Dissolve 2.68 g of compound M21-2 in 40 mL THF, add dropwise EtONa (1.19 g) ethanol (12 mL) solution at a temperature of -30 ^o C, and then naturally increase RT to react for 3 hours. TLC detected that the reaction was complete, and concentrated under reduced pressure to obtain a solid, which was beaten with MTBE (20 mL) to obtain 2.12 g of crude compound M21-3 as a solid, which could be directly used in the next reaction.

Preparation of intermediates M22-2 and M23-1:

Step 1: Preparation of compound M22-1

The M22-SM compound 1.29 g and 1.60 g were dissolved in 20 mL of M6 compound of tetrahydrofuran was added dropwise 1.12 g DIEA, 60 ^o C was stirred for 12 hours. TLC detected the completion of the reaction, concentrated under reduced pressure, added 50 mL of water, extracted with EtOAc (50 mL×3), combined the organic layers, dried over sodium sulfate and desolventized, purified by column chromatography to obtain 1.60 g of compound M22-1.

Step 2: Preparation of compounds M22-2 and M23-1

Under nitrogen atmosphere, 1.00 g M22-1 compound was dissolved in 30 mL of anhydrous dichloromethane, the temperature was lowered to -78 ^o C, was added dropwise DIBAL-H (1 M, 9.6 mL) in n-hexane. -78 ^o C reaction was stirred for 1 hour. Then slowly increase the temperature to -40 ^o C and continue the reaction for 2 hours. TLC detected the complete reaction of the raw materials. At 0 ^o C, slowly add 0.4 mL of water, then dropwise add sodium hydroxide aqueous solution (15%, 0.4 mL), and then add 1 mL of water. Warm to room temperature and stir for 15 minutes. Add sodium sulfate to dry, stir for ten minutes and filter. The filtrate was concentrated and purified by column chromatography to obtain 0.40 g of compound M22-2 and 0.11 g of compound M23-1.

Preparation of intermediate compound M24-4:

Step 1: Preparation of compound M24-2

The M24-1 compound 1.00 g was dissolved in 10 mL of dry acetonitrile was added 1.45 g of potassium carbonate, the reaction was stirred for 85 ^o C for 2 hours. The reaction was completed by TLC detection, the solvent was removed under reduced pressure, 50 mL of water was added, and then extracted with EA (50 mL×2), the organic layers were combined, dried with sodium sulfate and then desolventized, and purified by column chromatography to obtain 710 mg of compound M24-2 .

Step 2: Preparation of compound M24-3

Dissolve 710 mg of compound M24-2, 455 mg of compound M1-2, 895 mg of DIEA, 63 mg of Pd ₂ (dba) ₃ and 100 mg of Xantphos in 10 mL of dioxane. After nitrogen replacement for 3 times, 85 ^o C The reaction time is 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 84 mg of compound M24-3.

Step 3: Preparation of compound M24-4

Under nitrogen protection, 84 mg of compound M24-3 was dissolved in 2 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (0.85 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was slurried by adding 5 mL of methyl tert-butyl ether for 30 min, filtered, and the filter cake was vacuum-dried to obtain 60 mg of brown solid M24-4.

Preparation of intermediate compound M25-5:

Step 1: Preparation of compound M25-2

At 0 ^o C, add sodium hydroxide (546 mg) aqueous solution (25 mL) dropwise to the aqueous solution containing 1.00 g compound M25-1, 2.60 g elemental iodine and 1.70 g potassium iodide. The reaction was performed at room temperature for 2 h, and the reaction was complete as detected by TLC. Adjust the pH to neutral with saturated ammonium chloride solution. Then use sodium thiosulfate solution to remove the elemental iodine. It was extracted with methyl tert-butyl ether (50 mL×2), and the organic layers were combined, dried with sodium sulfate and desolvated. 1.45 g of compound M25-2 was obtained.

Step 2: Preparation of compound M25-3

1.45 g of compound M25-2, 1.13 g of methyl iodide and 1.10 potassium carbonate were dissolved in DMF (20 mL) and reacted at room temperature for 3 h. TLC detected that the reaction was complete. It was extracted with methyl tert-butyl ether (50 mL×2), and the organic layers were combined, dried with sodium sulfate and desolvated. After purification by column chromatography, 1.19 g of compound M25-2 was obtained.

Step 3: Preparation of compound M25-4

Dissolve 1.19 g of compound M25-3, 498 mg of compound M1-2, 1.07 g of DIEA, 76 mg of Pd ₂ (dba) ₃ and 120 mg of Xantphos in 10 mL of dioxane. After nitrogen replacement for 3 times, 85 ^o C The reaction time is 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 837 mg of compound M25-4.

Step 4: Preparation of compound M25-5

Under nitrogen protection, 870 mg of compound M25-4 was dissolved in 8 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (0.46 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was slurried by adding 5 mL of methyl tert-butyl ether for 30 min, filtered, and the filter cake was vacuum-dried to obtain 623 mg of brown solid M25-5.

Preparation of intermediate compound M26-3

Step 1: Preparation of compound M26-2

Dissolve 1.00 g of compound M26-1, 578 mg of compound M1-2, 1.24 g of DIEA, 88 mg of Pd ₂ (dba) ₃ and 139 mg of Xantphos in 10 mL of dioxane. After nitrogen replacement for 3 times, 85 ^o C The reaction time is 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 992 mg of compound M26-2.

Step 2: Preparation of compound M26-3

Under the protection of nitrogen, 992 mg of compound M26-2 was dissolved in 10 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (1.5 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was slurried by adding 10 mL of methyl tert-butyl ether for 30 min, filtered, and the filter cake was vacuum-dried to obtain 504 mg of brown solid M26-3.

Preparation of intermediate compound M27-11:

Step 1: Preparation of compound M27-3

10.00 g of compound M27-1 and 7.59 g of compound M27-2 were dissolved in 170 mL of chloroform, 350 mg of iodine was added, and the reaction was stirred at room temperature for 24 hours. TLC detects that the reaction is complete. Pour the compound into an aqueous sodium thiosulfate solution (100 mL, 0.4 mol/L), add an aqueous sodium hydroxide solution (70 mL, 40%), and extract with chloroform (150 mL×2). The organic layers were combined, dried over sodium sulfate, and desolventized, and recrystallized with dichloromethane/n-hexane to obtain 8.77 g of compound M27-3.

Step 2: Preparation of compound M27-5

Dissolve 8.77 g of compound M27-3 in 300 mL of anhydrous tetrahydrofuran, lower the temperature to -30 ^o C, add dropwise n-butyl lithium (2.5 M, 16 mL) in n-hexane, and keep stirring for 1 hour. Then 8.15 g of compound M27-4 was dissolved in 30 mL of tetrahydrofuran and slowly added dropwise to the reaction solution. After the addition, the temperature was raised to -10 ^o C and the reaction was continued for 2.5 hours. The reaction solution was quenched with 200 mL saturated brine, extracted with EtOAc (300 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 11.06 g of compound M27-5.

Step 3: Preparation of compound M27-7

11.06 g of compound M27-5 was dissolved in a mixed solution of water (40 mL) and dichloromethane (200 mL), and then 3.17 g of pyridine, 12.83 g of M27-6 and 0.86 g of tetrabutylammonium bromide were sequentially added. Stir at room temperature for 24 hours. The reaction solution was quenched with 100 mL saturated brine, extracted with DCM (100 mL×3), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 5.46 g of compound M27-7.

Step 4: Preparation of compound M27-8

Dissolve 5.46 g of compound M27-7 and 2.84 g of potassium tert-butoxide in 85 mL of anhydrous tetrahydrofuran, replace with nitrogen three times, and react in a microwave at 70 ^o C for 5 minutes. The solvent was removed under reduced pressure, 100 mL of saturated brine was added, and then extracted with EtOAc (100 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 2.91 g of compound M27-8.

Step 5: Preparation of compound M27-9

Dissolve 1.30 g compound M27-8, 1.56 g (R)-(+)-tert-butylsulfinamide in a mixed solution of tetraethyl titanate (10 mL) and anhydrous tetrahydrofuran (2 mL), and replace with nitrogen 3 times, 90 ^o C for 8 hours. Stop heating, pour the reaction solution into 200 mL ice water, then add 150 mL ethyl acetate, and stir for 1 hour. Filter to remove the filter residue. The filtrate was extracted with ethyl acetate (100 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 1.90 g of compound M27-9.

Step 6: Preparation of compound M27-10

The M27-9 compound 1.30 g was dissolved in 25 mL of anhydrous tetrahydrofuran at -20 ^o C was added portionwise 344 mg of sodium borohydride was slowly raised to room temperature and stirred for 3 hours at room temperature. At 0 ^o C, was added 100 mL of water, extracted with EtOAc (80 mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure, purified by column chromatography to give 0.94 g compound M27-10.

Step 7: Preparation of compound M27-11

0.94 g of compound M27-10 was dissolved in 10 mL of dichloromethane, and then 2.62 g of trifluoroacetic acid was added dropwise, and the mixture was stirred at room temperature for 1 hour. The reaction solution cooled to 0 ^o C, then saturated aqueous sodium bicarbonate was added to adjust the pH alkaline. Extract with a mixed solution of EtOAc:THF=1:1 (60 mL×4), combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. 692 mg of compound M27-11 was obtained.

Example 19 Preparation of compound 19:

Step 1: Preparation of compound 19-2

The compound 19-1 1.00 g was dissolved in 10 mL of dry acetonitrile was added 1.45 g of potassium carbonate, the reaction was stirred for 85 ^o C for 2 hours. The reaction was completed by TLC detection, the solvent was removed under reduced pressure, 50 mL of water was added, and then extracted with EA (50 mL×2), the organic layers were combined, dried with sodium sulfate and then desolventized, purified by column chromatography to obtain 720 mg of compound 19-2 .

Step 2: Preparation of compound 19-3

Dissolve 700 mg of compound 19-2, 450 mg of compound M1-2, 880 mg of DIEA, 63 mg of Pd ₂ (dba) ₃ and 100 mg of Xantphos in 10 mL of dioxane. After nitrogen replacement 3 times, 85 ^o C The reaction time is 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 271 mg of compound 19-3.

Step 3: Preparation of compound 19-4

Under nitrogen protection, 270 mg of compound 19-3 was dissolved in 5 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (2.0 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected the completion of the reaction. The reaction solution was concentrated under reduced pressure, and the residue was added to 5 mL DCM to make a slurry for 30 min. After filtration, the filter cake was washed with DCM (5 mL×3). The filter cake was vacuum-dried to obtain 154 mg of brown solid compound 19-4 .

Step 4: Preparation of compound 19-5

Dissolve 44 mg of compound 19-4, 40 mg of compound M22-2, 4 mg of Pd ₂ (dba) ₃ , 5 mg of Xantphos and 21 mg of DIEA in 2 mL of dioxane, replace with nitrogen 3 times, and heat to Reaction at 100 ^o C for 3 hours, followed by TLC. The solvent was removed under reduced pressure, 30 mL of water was added, and the mixture was extracted with dichloromethane (30 mL×2). The organic layers were combined, dried with sodium sulfate and then desolventized, and purified by column chromatography to obtain 28 mg of compound 19-5.

Step 5: Preparation of compound 19

Dissolve 28 mg of compound 19-5 in 1 mL of dioxane, add hydrochloric acid methanol solution (2 N, 0.4 mL) dropwise, and follow the reaction by TLC. After the reaction was completed, the solvent was removed under reduced pressure, and after solidification with a small amount of n-hexane, the n-hexane was poured out, 0.5 mL of water was added to dissolve the solid, and 2 drops of saturated sodium bicarbonate aqueous solution were added dropwise. The solid was obtained by filtration, and the solid was washed with a small amount of water and dried under vacuum. 15.7 mg of compound 19 was obtained as a pale yellow solid.

[M+H ⁺ ]=467.29

¹ H NMR (500 MHz, DMSO- d ₆ ,): δ 8.01 (s, 1H), 7.31 (d, J = 6.6 Hz, 1 H), 7.22-7.08 (m, 5 H), 6.91 (t, J = 7.0 Hz, 1 H), 5.35 (t, J = 5.0 Hz, 1 H), 4.48 (d, J = 5.2 Hz, 2 H), 3.85 (s, 3 H), 3.68 -3.64(m, 2 H ), 3.33-3.30 (m, 1 H), 3.10-3.00 (m, 3 H), 2.64-2.59 (m, 1 H), 1.91-1.86 (m, 1 H), 1.80-1.76 (m, 1 H) ), 1.51 (d, J = 12.8 Hz, 1 H), 1.13-1.10 (m, 1H).

Preparation of compound 20:

Step 1: Preparation of compound 20-2

318mg and 500mg of the compound M1-2 Compound 20-1 was dissolved in 6mL of dioxane was added _{48mg Pd 2 (dba) 3,} 76mg Xantphos and 684mg DIPEA, purged with nitrogen three times, the reaction solution was raised to 90 ^o C the reaction was stirred for 5hrs . TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 569 mg of compound 20-2.

Step 2: Preparation of compound 20-3

The compound 20-2 was dissolved in 569mg in 7mL THF, -30 ^o C solution of EtONa (1.3 mL, 20% in EtOH) at, RT reaction was stirred for 3hrs. TLC detected the completion of the reaction, concentrated under reduced pressure, added 10 mL DCM and stirred for 30 mins, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), and drained to obtain 380 mg of compound 20-3.

Step 3: Preparation of compound 20-4

35 mg of compound 20-3 and 70 mg of compound M22-2 were dissolved in 1 mL of dioxane, and 7 mg of Pd ₂ (dba) ₃ , 8 mg of Xantphos and 37 mg of DIPEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 55 mg of compound 20-4.

Step 4: Preparation of compound 20

55 mg of compound 20-4 was dissolved in 1 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.25 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (1.5 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (1.5 mL), take the filter cake and vacuum dry to obtain 12 mg of light yellow solid Compound 20.

[M+H ⁺ ]=451.24.

¹ H NMR (500 MHz, DMSO-d6) δ 8.36 (s, 1H), 8.32 (d, J = 5.3 Hz, 1H), 7.38-7.33 (m, 1H), 7.24-7.17 (m, 3H), 6.82 (d, J = 5.3 Hz, 1H), 5.50 (d, J = 6.3 Hz, 1H), 4.53 (d, J = 4.6 Hz, 2H), 3.98 (s, 1H), 3.93-3.86 (m, 2H) , 3.80 (s, 3H), 3.24-3.16 (m, 2H), 3.08 (d, J = 15.7 Hz, 1H), 2.71 (d, J = 15.7 Hz, 1H), 1.92-1.85 (m, 1H), 1.84-1.77 (m, 1H), 1.58-1.52 (m, 1H), 1.31-1.27 (m, 1H).

Example 21 Preparation of compound 21:

Step 1: Preparation of compound 21-1A

200 mg of compound M22-SM and 249 mg of compound M6 were dissolved in 2.5 mL of THF, and 137 mg of triethylamine was added. The reaction was stirred at RT for 4hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 222 mg of compound 21-1A.

Step 2: Preparation of compound 21-2A

222 mg of compound 21-1A and 109 mg of compound M1 were dissolved in 5 mL of dioxane, 19 mg of Pd ₂ (dba) ₃ and 25 mg of Xantphos were added. Nitrogen thrice, under nitrogen, the reaction was raised to 80 ^o C the reaction was stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 145 mg of compound 21-2A.

Step 3: Preparation of compound 21-3A

60 mg of compound 21-2A was dissolved in 1 mL of anhydrous THF. At 0 ^o C, was added portionwise 11mg LiAlH _4, 0 ^o C the reaction was stirred for 0.5hrs. TLC detects the completion of the reaction. Pour the reaction solution into 10 mL ice water, add DCM to extract (15 mL×3), combine the organic phases, wash with 15 mL saturated NaCl, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by TLC plate to obtain 12 mg White solid compound 21-3A.

Step 4: Preparation of compound 21

12 mg of compound 21-3A was dissolved in 0.5 mL of dioxane and 0.1 mL of MeOH, 2N HCl (0.1 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (0.5 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (0.5 mL), take the filter cake and vacuum dry to obtain 7 mg of white solid compound twenty one.

[M+H ⁺ ]=469.26.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.31 (s, 1H), 7.67 (d, J = 5.4 Hz, 1H), 7.31 (d, J = 6.8 Hz, 1H), 7.23-7.11 (m, 3H), 6.36 (s, 2H), 5.91 (d, J = 5.3 Hz, 1H), 5.48 (t, J = 5.9 Hz, 1H), 4.52 (d, J = 5.9 Hz, 2H), 3.94-3.83 ( m, 3H), 3.24-3.11 (m, 2H), 3.06 (d, J = 15.5 Hz, 1H), 2.63 (d, J = 15.3 Hz, 1H), 1.94-1.85 (m, 1H), 1.83-1.75 (m, 1H), 1.59-1.50 (m, 1H), 1.18-1.07 (m, 1H).

Example 22 Preparation of Compound 22:

Step 1: Preparation of compound 22-2

Under a nitrogen atmosphere, 600 mg of compound 22-1 and 414 mg of compound M1-2 were dissolved in 10 mL of dioxane, 105 mg of Pd ₂ (dba) ₃ , 166 mg of Xantphos, and 1.49 g of DIEA were added, and the temperature was raised to 85°C to react for 3 hrs. TLC detects that the reaction is complete, concentrate, add 50mL ethyl acetate and 50mL water to disperse, add 50mL ethyl acetate to the aqueous phase to extract, combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, the residue is purified by column chromatography to get off-white Solid 0.605g compound 22-2.

Step 2: Preparation of compound 22-3

0.605 g of compound 22-2 was dissolved in 5 mL of THF, EtONa (5.75 mL, 20% EtOH solution) was added dropwise at RT, and the reaction was stirred at RT for 3 hrs. TLC detected the completion of the reaction, concentrated under reduced pressure, added 15 mL of n-hexane and stirred for 30 mins, filtered the reaction solution, and washed the filter cake with n-hexane (15 mL×2) to obtain 660 mg of compound 22-3 as a beige solid.

Step 3: Preparation of compound 22-4

Under a nitrogen atmosphere, 50 mg of compound M22-2 and 22 mg of compound 22-3 were dissolved in 10 mL of dioxane, 9.28 mg of Pd ₂ (dba) ₃ , 11.73 mg of Xantphos and 26 mg of DIEA were added, and the temperature was raised to 100°C for 3 hrs. TLC detected that the reaction was complete, concentrated, and the residue was purified by preparative thin layer chromatography to obtain 8.4 mg of white compound 22-4.

Step 3: Preparation of compound 22

Under a nitrogen atmosphere, 8.4 mg of compound 22-5 was dissolved in 1.5 mL of dioxane and 0.5 mL of MeOH, and 2N HCl (0.32 mL, methanol solution) was added. The reaction was stirred at RT for 1 hrs. TLC detected that the reaction was complete and the reaction solution was reduced. Pressure concentration. Add H ₂ O (2 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (2 mL), take the filter cake and vacuum dry to obtain 1.2 mg of beige solid compound twenty two.

[M+H ⁺ ]=436.32.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.33 (s, 1H), 7.96 (d, J = 5.0 Hz, 1H), 7.31 (d, J = 7.0 Hz, 1H), 7.19-7.14 (m, 3H), 6.70 (s, 2H), 6.09 (d, J = 5.0 Hz, 1H), 5.47 (t, J = 5.0 Hz, 1H), 4.52 (d, J = 5.5 Hz, 2H), 3.89-3.82 ( m, 3H), 3.21-3.11 (m, 2H), 3.06 (d, J = 16.0 Hz, 1H), 2.63 (d, J = 15.5 Hz, 1H), 1.93-1.87 (m, 1H), 1.83-1.76 (m, 1H), 1.56-1.53 (m, 1H), 1.16-1.13 (m, 1H).

Example 23 Preparation of compound 23:

Step 1: Preparation of compound 21-A1

1.61 g of compound M22-SM and 2.00 g of compound M6 were dissolved in 20 mL of THF, 1.10 g of triethylamine was added, and the reaction was stirred at RT for 12 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (30 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 2.00 g of compound 21-A1.

Step 2: Preparation of compound 23-1

100 mg of compound 21-1A was dissolved in 1 mL of THF, 24 mg of LiOH·H ₂ O in H ₂ O (1 mL) was added dropwise, and the reaction was stirred at RT for 12 hrs. The reaction was completed by TLC. The reaction solution was adjusted to pH=7 with 1N HCl solution, and extracted with DCM (10mL×3). The organic phases were combined, washed with 15mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was passed through the column Analytical purification yielded 94 mg of compound 23-1.

Step 3: Preparation of compound 23-3

94 mg of compound 23-1 was dissolved in 1.5 mL of anhydrous DMF. At 0 ^o C, was added 46mg EDCI, 33mg HOBT, 24mg TEA and 17mg compound 23-2. The reaction was stirred at RT for 5hrs under nitrogen protection. TLC detected the completion of the reaction. The reaction solution was poured into 5mL water, extracted with EtOAc (15mL×2), combined the organic phases, washed three times with 15mL saturated NaCl, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography. 65 mg of compound 23-3.

Step 4: Preparation of compound 23-4

23-3 The compound of 65mg was dissolved in 1.8mL THF was added dropwise at _{^{-78 o C MgBrCH 3 (3M,}} 0.196mL) the reaction solution was raised to 0 ^o C the reaction was stirred for 1hrs. The reaction was completed by TLC detection. The reaction was quenched with saturated NH ₄ Cl (8 mL), extracted with EtOAc (15 mL×2), and the organic phases were combined, washed with 15 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was subjected to TLC plate Purified to obtain 10 mg of compound 23-4.

Step 5: Preparation of compound 23-5

10 mg of compound 23-4 and 6 mg of compound M1 were dissolved in 0.5 mL of dioxane, and 1 mg Pd ₂ (dba) ₃ , 1 mg Xantphos and 5 mg DIPEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 90 ^o C the reaction was stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (5 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by TLC plate to obtain 8 mg of compound 23-5.

Step 6: Preparation of compound 23

8 mg of compound 23-5 was dissolved in 0.4 mL dioxane and 0.1 mL MeOH, 2N HCl (0.1 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (0.5 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (0.5 mL), take the filter cake and vacuum dry to obtain 2.1 mg of yellow solid Compound 23.

[M+H ⁺ ]=481.23.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.41 (s, 1H), 7.70 (d, J = 5.3 Hz, 1H), 7.30 (d, J = 6.8 Hz, 1H), 7.22-7.13 (m, 3H), 6.37 (s, 2H), 5.97 (d, J = 5.4 Hz, 1H), 3.90-3.78 (m, 3H), 3.24 (q, J = 13.5, 13.0 Hz, 2H), 3.07 (d, J = 15.4 Hz, 1H), 2.65 (d, J = 15.3 Hz, 1H), 2.56 (s, 3H), 1.85-1.78 (m, 1H), 1.76-1.69 (m, 1H), 1.56-1.50 (m, 1H), 1.18-1.11 (m, 1H).

Example 24: Preparation of Compound 24:

Step 1: Preparation of compound 24-6

90 mg of compound M16-5 and 100 mg of compound M22-2 were dissolved in 1.5 mL of dioxane, and 5 mg of Pd ₂ (dba) ₃ , 6 mg of Xantphos and 26 mg of DIPEA were added. Nitrogen thrice, under nitrogen, the reaction was raised to 100 ^o C the reaction was stirred for 5hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 84 mg of compound 24-6.

Step 2: Preparation of compound 24

84 mg of compound 24-6 was dissolved in 1.5 mL of dioxane and 0.5 mL of MeOH, 2N HCl (0.32 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. The residue was dissolved by adding H ₂ O (2 mL), the solution was adjusted to pH=8 with saturated NaHCO ₃ , the solid precipitated out, filtered, the filter cake was washed with H ₂ O (2 mL), and the filter cake was vacuum-dried to obtain 55 mg of white solid 24.

[M+H ⁺ ]=549.22.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.51 (s, 1H), 8.35 (s, 1H), 7.76 (d, J = 5.4 Hz, 1H), 7.36 (d, J = 2.0 Hz, 2H) , 7.25-7.16 (m, 3H), 6.16 (d, J = 5.3 Hz, 1H), 6.09 (d, J = 1.9 Hz, 1H), 5.49 (t, J = 5.9 Hz, 1H), 4.54 (d, J = 5.7 Hz, 2H), 3.97 (s, 1H), 3.95-3.87 (m, 2H), 3.56 (s, 3H), 3.23-3.16 (m, 2H), 3.08 (d, J = 15.7 Hz, 1H ), 2.71 (d, J = 15.7 Hz, 1H), 1.93-1.85 (m, 1H), 1.85 -1.78 (m, 1H), 1.58-1.52 (m, 1H), 1.24-1.23 (m, 1H).

Example 25 Preparation of compound 25:

Step 1: Preparation of compound 25-2

The compound 500mg and 186mg M20-SM 25-1 compound was dissolved in 5 mL of DMSO was purged with nitrogen three times, under nitrogen, the reaction was raised to 70 ^o C the reaction was stirred for 4hrs. The reaction was completed by TLC detection. The reaction solution was poured into 20mL water, extracted with EtOAc (30mL×2), and the organic phases were combined, washed three times with 30mL saturated NaCl, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography. 490 mg of white solid compound 25-2.

Step 2: Preparation of compound 25-3

Dissolve 490 mg of compound 25-2 and 181.43 mg of compound M1-2 in 6 mL of dioxane, add 28 mg of Pd ₂ (dba) ₃ , 44 mg of Xantphos and 390 mg of DIPEA, replace with nitrogen three times, and raise the reaction solution to 90 ^o C and stir to react. 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 478 mg of compound 25-3.

Step 3: Preparation of compound 25-4

The compound 25-3 was dissolved in 478mg of 5mL THF, -30 ^o C was added dropwise EtONa (2mL, 20% in EtOH) at, RT reaction was stirred for 4hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 20 mL DCM and stirred for 30 mins, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), and drained to obtain 449 mg of compound 25-4.

Step 4: Preparation of compound 25-5

Dissolve 32 mg of compound 25-4 and 50 mg of compound M22-2 in 1 mL of dioxane, and add 5 mg of Pd ₂ (dba) ₃ , 6 mg of Xantphos and 26 mg of DIPEA. Nitrogen thrice, under nitrogen, the reaction was raised to 100 ^o C the reaction was stirred for 3hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 25 mg of compound 25-5.

Step 5: Preparation of compound 25

25 mg of compound 25-5 was dissolved in 0.7 mL of dioxane and 0.2 mL of MeOH, 2N HCl (0.1 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. The residue was dissolved by adding H ₂ O (1 mL), the solution was adjusted to pH=8 with saturated NaHCO ₃ , the solid precipitated out, filtered, the filter cake was washed with H ₂ O (1 mL), and the filter cake was vacuum dried to obtain 15 mg of white solid compound 25.

[M+H ⁺ ]=539.34.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.34 (s, 1H), 7.99 (d, J = 5.3 Hz, 1H), 7.31 (d, J = 6.9 Hz, 1H), 7.20-7.12 (m, 3H), 6.40 (d, J = 5.4 Hz, 1H), 5.47 (t, J = 5.9 Hz, 1H), 4.53 (d, J = 5.8 Hz, 2H), 3.93-3.88(m, 3H), 3.75- 3.72 (m, 4H), 3.23-3.12 (m, 6H), 3.06 (d, J = 15.6 Hz, 1H), 2.62 (d, J = 15.3 Hz, 1H), 1.93-1.86 (m, 1H), 1.83 -1.76 (m, 1H), 1.58-1.52 (m, 1H), 1.17-1.11 (m, 1H).

Example 26 Preparation of compound 26:

Step 1: Preparation of compound 26-2

The 0.50 g and 0.39 g compound compound M14 M22-SM was dissolved in 10 mL of tetrahydrofuran was added dropwise 0.26g DIEA, 60 ^o C was stirred for 12 hours. TLC detected the completion of the reaction, concentrated under reduced pressure, added 20 mL of water, extracted with EtOAc (20 mL×3), combined the organic layers, dried over sodium sulfate and desolventized, purified by column chromatography to obtain 0.58 g of compound 26-2.

Step 2: Preparation of compound 26-3

Under nitrogen atmosphere, 0.30 g Compound 26-2 was dissolved in 10 mL of anhydrous dichloromethane, the temperature was lowered to -78 ^o C, was added dropwise DIBAL-H (1 M, 2.6 mL) in n-hexane. -78 ^o C reaction was stirred for 1 hour. Then slowly increase the temperature to -40 ^o C and continue the reaction for 2 hours. TLC detects that the reaction is complete. At 0 ^o C, slowly add 0.1 mL of water, then dropwise sodium hydroxide aqueous solution (15%, 0.1 mL), and then add 0.25 mL of water. Warm to room temperature and stir for 15 minutes. Add anhydrous sodium sulfate to dry, stir for ten minutes and filter. The filtrate was concentrated and purified by column chromatography to obtain 0.21 g of compound 26-3.

Step 3: Preparation of compound 26-4

The compound 80mg 26-3,52mg Compound M-1 was dissolved in 4mL dioxane was added _3, 16mg Xantphos and _{130mg DIEA 13mg Pd 2 (dba)} , and then 100 ^o C the reaction 20 hrs. LCMS detected that the reaction was complete, the reaction solution was filtered, concentrated under reduced pressure, and the residue was purified with a preparation plate to obtain 35 mg of yellow solid compound 26-4.

Step 4: Preparation of compound 26

35 mg of compound 26-4 was dissolved in 2 mL of dioxane, and methanol solution of hydrochloric acid (2 N, 0.2 mL) was added dropwise, and the reaction was followed by TLC. After the reaction was completed, the solvent was removed under reduced pressure, and after solidification with a small amount of n-hexane, the n-hexane was poured out, 1.0 mL of water was added to dissolve the solid, and then saturated aqueous sodium bicarbonate solution was added dropwise to adjust pH=9-10. The solid was obtained by filtration, and the solid was washed with a small amount of water and dried in vacuo to obtain 20.8 mg of compound 26 as a pale yellow solid.

[M+H] ⁺ : 537.16

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.32 (s, 1H), 7.70 – 7.64 (m, 2H), 7.53 (d, J = 7.8 Hz, 1H), 7.42 (d, J = 7.8 Hz, 1H), 6.35 (s, 2H), 5.92 (d, J = 5.4 Hz, 1H), 5.49 (t, J = 5.9 Hz, 1H), 4.53 (d, J = 5.8 Hz, 2H), 4.01-3.84 ( m, 3H), 3.23-3.13 (m, 3H), 2.75-2.72 (m, 1H), 1.97-1.90 (m, 1H), 1.83-1.80 (m, 1H), 1.60-1.57 (m, 1H), 1.13-1.10 (m, 1H).

Example 27 Preparation of compound 27:

Step 1: Preparation of compound 27-1

The compound 400mg M23-1 was dissolved in 20.0mL THF and cooled in an ice bath to 0 ^o C, methylmagnesium bromide (2.0M, 2ml) was slowly added to the solution, the reaction was warmed to room temperature stirred for 0.5hrs. The reaction was completed by TLC detection. Saturated ammonium chloride aqueous solution (5mL) was slowly added to the reaction solution to quench the reaction, EA (10mL*3) was added for extraction, the organic phases were combined, washed with saturated brine (15mL), dried with anhydrous sodium sulfate, and filtered The filtrate was spin-dried, and the residue was purified by column chromatography to obtain 60 mg of white solid compound 27-1.

Step 2: Preparation of compound 27-2

60 mg of compound 27-1 and 44 mg of compound M-1 were dissolved in 5 mL of dioxane, and 11 mg of Pd ₂ (dba) ₃ , 14 mg of Xantphos, and 46 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction is raised to 100 ^o C and the reaction is stirred for 18 hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 21 mg of compound 27-2.

Step 3: Preparation of compound 27

21 mg of compound 27-2 was dissolved in 1.5 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.5 mL, methanol solution) was added, and the reaction was stirred at RT for 0.5 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (0.5 mL) to dissolve the residue, adjust the solution to pH=8-9 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (0.5 mL), dissolve the solid with DCM, and dry it It was concentrated in vacuo and purified by a preparation plate to obtain 16.3 mg of white solid 27.

[M+H ⁺ ]=483.21.

¹ H ^NMR (500 MHz, DMSO- d ₆ ) δ 8.32 (s, 1H), 7.67 (d, J = 5.5 Hz, 1H), 7.31 (d, J = 7.0 Hz, 1H), 7.19-7.13 (m, 3H), 6.37 (s, 2H), 5.95 (d, J = 5.5 Hz, 1H), 5.36 (m, 1H), 4.91-4.88 (m,1H), 3.97-3.84 (m,2H), 3.71-3.63 (m,1H), 3.21-3.02(m,3H), 2.6-2.59(m, 1H), 1.89-1.75 (m,2H), 1.56-1.53 (m, 1H), 1.39 (d, J = 6.0 Hz ,3H), 1.17-1.13 (m,1H).

^Via different starting materials and appropriate reagents, Example 26 using a method similar to Table 3 were prepared in the foregoing embodiments the compound 63 and 28-57.

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((5-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇 549.19 29

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-(二甲基氨基)-3-氟吡啶-4-基)硫基)吡嗪-2-基)甲醇 481.26 30

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-吡啶]-1'-基)-6-((3-氟-2-(甲基氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇 467.25 31

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-(二氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲基 470.25 32

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((3-氯-2-(二甲基氨基)吡啶-4-基)硫基)吡嗪-2-基)甲基 497.24 33

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((3-氟-2-甲氧基苯基)硫基)吡嗪-2-基)甲醇 467.26 34

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((3-氯-5-氟-2-甲氧基苯基)硫基)吡嗪-2-基)甲醇 501.25 35

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-(喹啉-4-基硫基)吡嗪-2-基)甲醇 470.32 36

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲醇 488.30 37

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2,3-二氯苯基)硫基)吡嗪-2-基)甲醇 487.17 38

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲醇 488.25 39

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-氨基-3-氟吡啶-4-基)硫基)吡嗪-2-基)甲基 453.27 40

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((3-氯-2-甲氧基吡啶-4-基)硫基)吡嗪-2-基)甲醇 484.19 41

(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氟-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-基)甲醇 487.21 42

(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-甲氧基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-基)甲醇 499.23 43

(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-5,6-二甲基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-基)甲醇 530.21 44

(R)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(3-氨基-3H-螺[苯并呋喃-2,4'-呱啶]-1'-基)吡嗪-2-基)甲基 471.23 45

(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羥基甲基)吡嗪-2-基)-1,3-二氫螺[茚-2,4'-呱啶]-4-腈 494.21 46

(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氯-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-基)甲醇 503.14 47

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-(吡啶-4-基硫基)吡嗪-2-基)甲醇 420.26 48

(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(5-氨基-5,7-二氫螺[環戊烯并[b]吡啶-6,4'-呱啶]-1'-基)吡嗪-2-基)甲基 470.15 49

(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-(三氟甲基)-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-基)甲醇 537.14 50

(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羥基甲基)吡嗪-2-基)-1,3-二氫螺[茚-2,4'-呱啶]-6-醇 485.14 51

(S)-4-((5-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-(羥基甲基)吡嗪-2-基)硫基)嘧啶-2-醇 437.17 52

(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羥基甲基)吡嗪-2-基)-7-氟-1,3-二氫螺[茚-2,4'-呱啶]-4-醇 503.14 53

(S)-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2,3-二氫-[1,4] 二氧雜環己并[2,3-b]吡啶-8-基)硫基)吡嗪-2-基)甲醇 478.18 54

(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羥基甲基)吡嗪-2-基)-1,3-二氫螺[茚-2,4'-呱啶]-6-腈 494.15 55

(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羥基甲基)吡嗪-2-基)-4-氟-1,3-二氫螺[茚-2,4'-呱啶]-7-腈 512.14 56

(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-7-(三氟甲基)-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)吡嗪-2-基)甲基 537.14 57

(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羥基甲基)吡嗪-2-基)-1,3-二氫螺[茚-2,4'-呱啶]-7-醇 485.14 63

(S)-2-(3-(1-氨基-1,3-二氫螺[茚-2,4'-呱啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)丙-2-醇 497.18 table 3 Numbering Chemical structure Chinese name LC-MS [M+H ⁺ ] 28

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((5-chloro-2-((1 -Methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol 549.19 29

(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-(dimethylamino)- 3-fluoropyridin-4-yl)thio)pyrazin-2-yl)methanol 481.26 30

(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-fluoro-2-(methylamino )Pyridin-4-yl)thio)pyrazin-2-yl)methanol 467.25 31

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-(difluoromethyl)pyridine -3-yl)thio)pyrazin-2-yl)methyl 470.25 32

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-chloro-2-(dimethyl (Amino)pyridin-4-yl)thio)pyrazin-2-yl)methyl 497.24 33

(S)-(3-(1-Amino-1,3-dihydrospiro[inden-2,4'-piperidine]-1'-yl)-6-((3-fluoro-2-methoxy (Phenyl)thio)pyrazin-2-yl)methanol 467.26 34

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-chloro-5-fluoro-2 -Methoxyphenyl)thio)pyrazin-2-yl)methanol 501.25 35

(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(quinolin-4-ylthio)pyridine (Azin-2-yl) methanol 470.32 36

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-(trifluoromethyl)pyridine -3-yl)thio)pyrazin-2-yl)methanol 488.30 37

(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorophenyl) (Thio)pyrazin-2-yl)methanol 487.17 38

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-(trifluoromethyl)pyridine -3-yl)thio)pyrazin-2-yl)methanol 488.25 39

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-amino-3-fluoropyridine- 4-yl)thio)pyrazin-2-yl)methyl 453.27 40

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methoxy (Pyridin-4-yl)thio)pyrazin-2-yl)methanol 484.19 41

(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-fluoro-1,3-dihydrospiro[indene-2,4 '-Pepidine)-1'-yl)pyrazin-2-yl)methanol 487.21 42

(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-methoxy-1,3-dihydrospiro(indene-2 ,4'-Pepidine)-1'-yl)pyrazin-2-yl)methanol 499.23 43

(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro(indene -2,4'-piperidine)-1'-yl)pyrazin-2-yl)methanol 530.21 44

(R)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(3-amino-3H-spiro[benzofuran-2,4'-piperidine]- 1'-yl)pyrazin-2-yl)methyl 471.23 45

(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1, 3-Dihydrospiro[indene-2,4'-piperidine]-4-carbonitrile 494.21 46

(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2,4 '-Pepidine)-1'-yl)pyrazin-2-yl)methanol 503.14 47

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-(pyridin-4-ylthio)pyrazine -2-base) methanol 420.26 48

(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(5-amino-5,7-dihydrospiro[cyclopenteno[b]pyridine- 6,4'-Pepidine)-1'-yl)pyrazin-2-yl)methyl 470.15 49

(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-(trifluoromethyl)-1,3-dihydrospiro[ Indene-2,4'-piperidine)-1'-yl)pyrazin-2-yl)methanol 537.14 50

(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1, 3-Dihydrospiro[indene-2,4'-piperidine]-6-ol 485.14 51

(S)-4-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazine- 2-yl)thio)pyrimidin-2-ol 437.17 52

(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-7- Fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-4-ol 503.14 53

(S)-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2,3-dihydro-[1 ,4] Dioxan[2,3-b]pyridin-8-yl)thio)pyrazin-2-yl)methanol 478.18 54

(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1, 3-Dihydrospiro[indene-2,4'-piperidine]-6-nitrile 494.15 55

(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-4- Fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-7-nitrile 512.14 56

(S)-(6-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-7-(trifluoromethyl)-1,3-dihydrospiro[ Indene-2,4'-pyridine)-1'-yl)pyrazin-2-yl)methyl 537.14 57

(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1, 3-Dihydrospiro[indene-2,4'-piperidine]-7-ol 485.14 63

(S)-2-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloro (Pyridin-4-yl)thio)pyrazin-2-yl)propan-2-ol 497.18

The NMR data of compounds 30, 33, 35, 36, 37, 41, 42, 45, 46, 47 are as follows:

[M+H ⁺ ]=467.25

¹ H NMR (500 MHz, DMSO- d ₆ ): δ 8.25 (s, 1H), 7.67 (d, J = 5.4 Hz, 1H), 7.31 (d, J = 6.9 Hz, 1H), 7.20-7.12 (m , 3H), 6.69 (d, J = 5.1 Hz, 1H), 6.12 (t, J = 4.5 Hz, 1H), 5.46 (s, 1H), 4.50 (s, 2H), 3.85-3.77 (m, 3H) , 3.18-3.11 (m, 2H), 3.03 (d, J = 15.6 Hz, 1H), 2.82 (d, J = 4.5 Hz, 3H), 2.61 (d, J = 15.6 Hz, 1H), 1.88 (m, 1H), 1.83-1.74 (m, 1H), 1.55-1.52 (m, 1H), 1.14-1.11 (m, 1H). (Compound 30)

[M+H ⁺ ]=467.26

¹ H NMR (500 MHz, DMSO- d ₆ ): δ 8.11 (s, 1 H), 7.31 (d, J = 6.5 Hz, 1 H), 7.25-7.09 (m, 4H), 7.08-7.04 (m, 1H), 6.89 (d, J = 7.8 Hz, 1 H), 5.39 (t, J = 5.6 Hz, 1 H), 4.50 (d, J = 5.5 Hz, 2H), 3.85 (s, 3 H), 3.75 -3.70 (m, 2H), 3.14-3.02 (m, 3 H), 2.61 (d, J = 15.6 Hz, 2 H), 1.91-1.81 (m, 1 H), 1.81-1.76 (m, 1 H) , 1.53 (d, J = 12.9 Hz, 1 H), 1.13 (d, J = 13.3 Hz, 1 H). (Compound 33)

[M+H ⁺ ]=470.32

¹ H NMR (500 MHz, DMSO- d ₆ ): δ 8.66 (d, J = 4.7 Hz, 1 H), 8.36 (s, 1 H), 8.20 (d, J = 8.3 Hz, 1 H), 8.05 ( d, J = 8.4 Hz, 1 H), 7.84 (t, J = 7.6 Hz, 1 H), 7.71 (t, J = 7.5 Hz, 1 H), 7.32 (d, J = 6.6 Hz, 1 H), 7.20-7.16 (m, 3 H), 7.04 (d, J = 4.6 Hz, 1 H), 5.49 (t, J = 5.8 Hz, 1 H), 4.54 (d, J = 5.7 Hz, 2 H), 4.01 -3.80 (m, 3H), 3.21-3.18 (m, 2 H), 3.07 (d, J = 15.7 Hz, 1 H), 2.62 (d, J = 15.7 Hz, 1H), 1.93-1.88 (m, 1 H), 1.83-1.78 (m, 1 H), 1.57-1.54 (m, 1 H), 1.17-1.13 (m, 1 H). (Compound 35)

[M+H] ⁺ : 488.3

¹ H NMR (500 MHz, DMSO- d ₆ ) δ : 8.58-8.56(m, 1H), 8.25 (s, 1H), 7.81-7.65 (m, 1H), 7.62-7.60 (m, 1H), 7.32- 7.29 (m, 1H), 7.17-7.13 (m, 3H), 5.41 (t, J = 5.6 Hz, 1H), 4.49 -4.47(m, 2H), 3.85-3.75 (m, 2H), 3.20-3.10 ( m, 2H), 3.05-3.02 (m, 1H), 2.63-2.55 (m, 1H), 1.96-1.67 (m, 1H), 1.55-1.52 (m, 1H), 1.18-1.08 (m, 1H). (Compound 36)

[M+H] ⁺ : 487.17

¹ H NMR (500 MHz, DMSO- d ₆ ) δ : 8.24 (s, 1H), 7.51 (d, J = 8.0 Hz, 1H), 7.34 – 7.25 (m, 2H), 7.17 (q, J = 7.6, 7.2 Hz, 3H), 6.99 (d, J = 8.0 Hz, 1H), 5.43 (t, J = 5.8 Hz, 1H), 4.51 (d, J = 5.8 Hz, 2H), 3.85-3.77 (m, 3H) , 3.20-3.10 (m, 2H), 3.05 (d, J = 15.6 Hz, 1H), 2.62 (d, J = 15.6 Hz, 1H), 2.02-1.68 (m, 4H), 1.55-1.53 (m, 1H) ), 1.15-1.13 (m, 1H). (Compound 37)

[M+H] ⁺ : 487.21

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.31 (s, 1H), 7.67 (d, J = 5.6 Hz, 1H), 7.21-7.18 (m, 1H), 7.09 (d, J = 9.0 Hz, 1H), 6.95 (t, J = 8.8 Hz, 1H), 6.36 (s, 2H), 5.99-5.88 (m, 1H), 5.49 (t, J = 5.9 Hz, 1H), 4.52 (d, J = 5.8 Hz, 2H), 3.92-3.85 (m, 3H), 3.21-3.01 (m, 3H), 2.63-2.57 (m, 1H), 1.99-1.89 (m, 1H), 1.84-1.74 (m, 1H), 1.58-1.55 (m, 1H), 1.13-1.10 (m, 1H). (Compound 41)

[M+H] ⁺ : 499.23

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.31 (s, 1H), 7.67 (d, J = 5.4 Hz, 1H), 7.07 (d, J = 8.2 Hz, 1H), 6.90 (m, 1H) , 6.70 (d, J = 7.9 Hz, 1H), 6.36 (s, 2H), 5.91 (d, J = 5.3 Hz, 1H), 5.48 (m, 1H), 4.52 (d, J = 4.9 Hz, 2H) , 3.94-3.76 (m, 3H), 3.73 (s, 3H), 3.20-3.10 (m, 2H), 2.98-2.95 (m, 1H), 2.64 -2.55 (m, 1H), 1.93-1.88 (m, 1H), 1.80-1.75 (m, 1H), 1.56-1.53 (m, 1H), 1.24-1.11 (m, 1H). (Compound 42)

[M+H] ⁺ : 494.21

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.32 (s, 1H), 7.69-7.61 (m, 3H), 7.41 (t, J = 7.6 Hz, 1H), 6.35 (s, 2H), 5.92 ( d, J = 5.4 Hz, 1H), 5.49 (t, J = 5.8 Hz, 1H), 4.53 (d, J = 5.8 Hz, 2H), 4.06-3.85 (m, 3H), 3.29-3.18 (m, 3H ), 2.86-2.83 (m, 1H), 1.91-1.80 (m, 2H), 1.61-1.59 (m, 1H), 1.22 – 1.15 (m, 1H). (Compound 45)

[M+H] ⁺ : 503.14

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.31 (s, 1H), 7.67 (d, J = 6.0 Hz, 1H), 7.32 (s, 1H), 7.24 – 7.16 (m, 2H), 6.36 ( s, 2H), 5.91 (d, J = 5.3 Hz, 1H), 5.49 (t, J = 5.9 Hz, 1H), 4.52 (d, J = 5.8 Hz, 2H), 3.92-3.86 (m, 3H), 3.21-3.11 (m, 2H), 3.05 (d, J = 15.7 Hz, 1H), 2.60 (d, J = 15.7 Hz, 1H), 1.93-1.88 (m, 1H), 1.80-1.75 (m, 1H) , 1.56 (m, 1H), 1.12-1.10 (m, 1H). (Compound 46)

[M+H] ⁺ : 420.26

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.41-8.36 (m, 2H), 8.32 (s, 1H), 7.32 (d, J = 6.8 Hz, 1H), 7.23-7.09 (m, 5H), 5.48 (t, J = 5.6 Hz, 1H), 4.53 (d, J = 5.6 Hz, 2H), 3.91-3.80 (m, 3H), 3.21-3.13 (m, 2H), 3.06 (d, J = 15.6 Hz) , 1H), 2.65 (d, J = 15.6 Hz, 1H), 1.93-1.77 (m, 2H), 1.55-1.53 (m, 1H), 1.18-1.15 (m, 1H). (Compound 47)

Example 60 Preparation of Compound 60:

Step 1: Preparation of compound 60-1

The compound of 13mg was dissolved in 0.5mL MeOH 21-2A was added dropwise 3mg LiOH · H ₂ O in H ₂ O (0.1mL) added and the reaction was warmed to 40 ^o C the reaction was stirred for 12hrs. TLC detects that the reaction is complete, adjust the reaction solution to pH=7 with 1N HCl solution, add DCM/MeOH=10/1 to extract (5mL×2), combine the organic phases, wash with 5mL saturated NaCl, dry with anhydrous sodium sulfate, and concentrate under reduced pressure The residue was purified by TLC plate to obtain 10 mg of compound 60-1.

Step 2: Preparation of compound 60-3

10mg compound 60-1 was dissolved in 0.3mL THF and 0.3mL DCM, 4mg EDCI, 3mg HOBT and 2mg TEA were added, and the reaction was stirred at RT for 1hrs. Another 3 mg of compound 60-2 was added. Replace with nitrogen for three times, and stir for 12hrs at RT under nitrogen protection. TLC detected that the reaction was complete, concentrated under reduced pressure, and the residue was purified by TLC plate to obtain 6 mg of compound 60-3.

Step 3: Preparation of compound 60

6 mg of compound 60-3 was dissolved in 0.5 mL of MeOH, 2N HCl (0.1 mL, methanol solution) was added, and the reaction was stirred at RT for 8 hrs. TLC detected that the reaction was complete and the reaction solution was concentrated under reduced pressure. Add H ₂ O (0.5 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (0.5 mL), take the filter cake and vacuum dry to obtain 2.2 mg of yellow solid Compound 60.

[M+H ⁺ ]=498.23.

Example 61 Preparation of Compound 61:

Step 1: Preparation of compound 61-3

100 mg of compound M23-1 and 57 mg of compound 61-2 were dissolved in 2 mL of tetrahydrofuran, 0.01 mL of tetrabutylammonium fluoride was added, and then reacted for 2 hours at RT. TLC detected the complete reaction of the raw materials, the reaction solution was spin-dried, and purified by column chromatography to obtain 61 mg of compound 61-3.

Step 2: Preparation of compound 61-4

Dissolve 61 mg of compound 61-3 and 50 mg of compound M1 in 1.5 mL of dioxane, add 10 mg Pd ₂ (dba) ₃ , 12 mg Xantphos and 42 mg DIEA, and then react at 100 ^o C for 4 hrs. LCMS detected that the reaction was complete, the reaction solution was filtered, concentrated under reduced pressure, and the residue was purified with a preparation plate to obtain 35 mg of white solid compound 61-4.

Step 3: Preparation of compound 61

35 mg of compound 61-4 was dissolved in 1.5 mL of dioxane and 0.5 mL of methanol, 0.14 mL of HCl in dioxane was added, and stirred at room temperature for 2 hours. LCMS detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, the residue was dissolved in 1 mL of water, saturated sodium bicarbonate solution was added dropwise to adjust pH=8, a solid was precipitated, filtered and dried to obtain 20.6 mg of white solid compound 61.

[M+H ⁺ ]=537.18.

Example 62 Preparation of Compound 62:

Step 1: Preparation of compound 62-2

100 mg of compound M22-2 and 72 mg of benzyl tributylammonium bromide were dissolved in a mixed solution of 4 mL of sodium hydroxide (50%) and 4 mL of dichloromethane. Add 26 mg of compound 62-1 dropwise at 0 ^o C and slowly warm to room temperature. TLC detected that the reaction was complete. The reaction was quenched by adding 30 mL of water, and then extracted with EA (50 mL). The organic layers were combined, dried with sodium sulfate and desolventized, and purified by column chromatography to obtain 49 mg of compound 62-2.

Step 2: Preparation of compound 62-3

Dissolve 49 mg of compound 62-2, 53 mg of compound M1, 25 mg of DIEA, 9 mg of Pd ₂ (dba) _3, and 10 mg of Xantphos in 2 mL of dioxane. After nitrogen replacement 3 times, 100 ^o C reaction 3 hrs. Stop heating, concentrate under reduced pressure, add 20 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 27 mg of compound 62-3.

Step 3: Preparation of compound 62

Dissolve 27 mg of compound 62-3 in 1 mL of dioxane, add hydrochloric acid methanol solution (2 N, 0.4 mL) dropwise, and follow the reaction by TLC. After the reaction was completed, the solvent was removed under reduced pressure, and after solidification with a small amount of n-hexane, the n-hexane was poured out, 0.5 mL of water was added to dissolve the solid, and 2 drops of saturated sodium bicarbonate aqueous solution were added dropwise. The solid was obtained by filtration, and the solid was washed with a small amount of water and dried under vacuum. 10.5 mg of compound 62 was obtained as a pale yellow solid.

¹ H NMR (500 MHz, DMSO- d ₆ ,): δ 8.35 (s, 1H), 7.67 (d, J = 5.4 Hz, 1 H), 7.31 (d, J = 6.5 Hz, 1 H), 7.18- 7.15 (m, 3 H), 6.36 (s, 2 H), 5.88 (d, J = 5.1 Hz, 1 H), 4.45 (s, 2 H), 3.93-3.86 (m, 3 H), 3.25-3.15 (m, 2 H), 3.06 (d, J = 15.7 Hz, 1 H), 2.63 (d, J = 15.3 Hz, 1 H), 1.92-1.88 (m, 1 H), 1.82-1.78 (m, 1 H), 1.55 (d, J = 12.9 Hz, 1 H), 1.16 (d, J = 13.8 Hz, 1 H).

Example 64 Preparation of Compound 64:

Step 1: Preparation of compound 64-1

The M22-SM compound 400mg and 552mg the compound M12 was dissolved in 8.0mL THF was added 349mg N, N- diisopropylethylamine, 40 ^o C the reaction was stirred for 16hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 420 mg of light yellow solid compound 64-1.

Step 2: Preparation of compound 64-2

420 mg of compound 64-1 was dissolved in 5 mL of anhydrous DCM. Under nitrogen, -78 ^o C was slowly added dropwise 3.8 mL 1.0 M DIBAL-H in hexane solution, -78 ^o C the reaction was stirred for 15 min. LCMS detected the complete reaction of the raw materials. 0.5mL water and 0.5mL NaOH (2.5M) aqueous solution were added dropwise to quench at low temperature. After warming to room temperature, 0.5mL water was added dropwise. After filtration, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. 385 mg of crude compound 64-2 was obtained.

Step 3: Preparation of compound 64-3

385 mg of compound 64-2 was dissolved in 5 mL of anhydrous THF. Under nitrogen, 0 ^o C was added 56mg NaBH _4, reaction was brought to room temperature stirred for 2hrs. LCMS detected the completion of the reaction. Add 5mL saturated ammonium chloride dropwise under ice bath, return to room temperature, concentrate in vacuo and spin dry THF, the aqueous phase was extracted with DCM (5mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure The residue was purified by Pre-TLC to obtain 130 mg of yellow solid compound 64-3.

Step 4: Preparation of compound 64-4

130 mg of compound 64-3 and 63 mg of compound 22-3 were dissolved in 5 mL of 1,4-dioxane, 12 mg of Pd ₂ (dba) ₃ , 15 mg of Xantphos, and 65 mg of N,N-diisopropylethylamine were added. Nitrogen thrice, under nitrogen, the reaction was warmed to 70 ^o C the reaction was stirred for 2hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by Pre-TLC to obtain 70 mg of yellow solid compound 64-4.

Step 4: Preparation of compound 64

70 mg of compound 64-4 was dissolved in 3.0 mL 1,4-dioxane and 0.6 mL MeOH, 2N HCl (0.3 mL, methanol solution) was added, and the reaction was stirred at room temperature for 2.0 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (0.5 mL) to dissolve the residue, adjust the solution to pH=8~9 with saturated NaHCO ₃ , solid precipitate, filter, filter cake solid is dissolved in DCM, add anhydrous sodium sulfate and dry, filter and concentrate in vacuo to obtain 50.2 mg of compound 64 as a pale yellow solid.

[M+H ⁺ ]=466.34.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.33 (s, 1H), 7.96 (d, J = 5.0 Hz, 1H), 7.08 (d, J = 8.0 Hz, 1H), 6.90-6.89 (m, 1H), 6.72-6.69 (m, 3H), 6.09 (d, J = 5.5 Hz, 1H), 5.50 (m, 1H), 4.52 (d, J = 2.5 Hz, 2H), 3.89-3.81 (m, 3H ), 3.73(s, 3H), 3.18-3.08 (m, 2H), 2.96 (d, J = 15.0 Hz, 1H), 2.54(d, J = 15.0 Hz, 1H), 1.93-1.88 (m, 1H) , 1.80-1.75(m, 1H), 1.55-1.53 (m, 1H), 1.13-1.10 (m, 1H).

Example 65 Preparation of compound 65:

Step 1: Preparation of compound 65-1

150 mg of compound M22-2 and 41 mg of compound 4-mercaptopyrimidine were dissolved in 5 mL of 1,4-dioxane, and 14 mg of Pd ₂ (dba) ₃ , 18 mg of Xantphos and 118 mg of N,N-diisopropylethylamine were added. Nitrogen thrice, under nitrogen, the reaction was warmed to 70 ^o C the reaction was stirred for 2hrs. LCMS and TLC detected the completion of the reaction, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by Pre-TLC to obtain 60 mg of yellow solid compound 65-1.

Step 2: Preparation of compound 65

60 mg of compound 65-1 was dissolved in 1.5 mL 1,4-dioxane and 0.5 mL MeOH, 2N HCl (0.3 mL, methanol solution) was added, and the reaction was stirred at room temperature for 1.0 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. The residue was dissolved by adding H ₂ O (0.5 mL), the solution was adjusted to pH=7-8 with saturated NaHCO ₃ , a solid was precipitated, filtered, the filter cake was washed with water, and dried to obtain 19.1 mg of light yellow solid 65.

[M+H ⁺ ]=421.27.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.91 (s, 1H), 8.51 (d, J = 5.5 Hz, 1H), 8.36 (s, 1H), 7.32 (d, J = 6.5 Hz, 1H) , 7.21-7.13 (m, 4H), 5.30 (t, J = 6.0 Hz, 1H), 4.53 (s, 2H), 3.93-3.86 (m, 3H), 3.33-3.16 (m, 2H), 3.07 (d , J = 15.0 Hz, 1H), 2.64(d, J = 15.5 Hz, 1H), 1.93-1.88 (m, 1H), 1.82-1.77 (m, 1H), 1.57-1.54 (m, 1H), 1.16- 1.13 (m, 1H).

Example 66 Preparation of compound 66:

Step 1: Preparation of compound 66-1

500 mg of compound M22-SM and 634 mg of compound M16 were dissolved in 9.0 mL of dioxane, and 655 mg of N,N-diisopropylethylamine was added. 70 ^o C the reaction was stirred for 2hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 444 mg of yellow solid compound 66-1.

Step 2: Preparation of compound 66-2

444 mg of compound 66-1 was dissolved in 10 mL of anhydrous DCM. Under nitrogen, -78 ^o C was slowly added dropwise 4.0 mL 1.0 M DIBAL-H in hexane solution, -78 ^o C the reaction was stirred for 0.5hrs. The reaction was completed by LCMS. 0.5mL water was added dropwise at low temperature, and 0.5mL 2.5 M NaOH aqueous solution was added dropwise to quench. After returning to room temperature, 0.5mL water was added dropwise. After filtration, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 380mg of crude compound 66 -2.

Step 3: Preparation of compound 66-3

380 mg of compound 66-2 was dissolved in 10 mL of anhydrous THF. Under nitrogen, 0 ^o C was added 60mg NaBH _4, reaction was brought to room temperature stirred for 0.5hrs. LCMS detected the completion of the reaction. The reaction was quenched by adding 5 mL of water dropwise under an ice bath. After returning to room temperature, the aqueous phase was extracted with DCM (5 mL×3), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was purified by column chromatography to obtain 300 mg of yellow solid compound 66-3.

Step 4: Preparation of compound 66-4

200 mg of compound 66-3 and 96 mg of compound 22-3 were dissolved in 5 mL of dioxane, and 35 mg of Pd ₂ (dba) ₃ , 44 mg of Xantphos, and 147 mg of N,N-diisopropylethylamine were added. Nitrogen thrice, under nitrogen, the reaction was raised to 70 ^o C the reaction was stirred for 2hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 160 mg of compound 66-4.

Step 5: Preparation of compound 66

50 mg of compound 66-4 was dissolved in 1.5 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.5 mL, methanol solution) was added, and the reaction was stirred at RT for 0.5 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (0.5 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (0.5 mL), dissolve the solid with DCM, dry and concentrate in vacuo , Purified by preparation plate to obtain 5.8 mg of white solid 66.

[M+H ⁺ ]=470.31.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.34 (s, 1H), 7.96 (d, J = 5.0 Hz, 1H), 7.32 (d, J = 8.0 Hz, 1H), 7.24 -7.21 (m, 2H), 6.72 (s, 2H), 6.08 (d, J = 5.5 Hz, 1H), 5.50 (t, J = 6.0 Hz, 1H), 4.52 (d, J = 6.0 Hz, 2H), 3.88-3.82 ( m, 3H), 3.18-3.09 (m, 2H), 3.06 (d, J = 16.0 Hz, 1H), 2.63 (d, J = 15.5 Hz, 1H), 1.92-1.86 (m, 1H), 1.81-1.76 (m, 1H), 1.56-1.53 (m, 1H), 1.14-1.13 (m, 1H).

Example 67 Preparation of compound 67:

Step 1: Preparation of compound 67-1

500 mg of compound M22-SM and 603 mg of compound M17 were dissolved in 9.0 mL of dioxane, and 655 mg of N,N-diisopropylethylamine was added. 70 ^o C the reaction was stirred for 2hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 453 mg of yellow solid compound 67-1.

Step 2: Preparation of compound 67-2

453 mg of compound 67-1 was dissolved in 10 mL of anhydrous DCM. Under nitrogen, -78 ^o C was slowly added dropwise 4.0 mL 1.0 M DIBAL-H in hexane solution, -78 ^o C the reaction was stirred for 0.5hrs. The reaction was completed by LCMS. 0.5mL water was added dropwise at low temperature, and 0.5mL 2.5 M NaOH aqueous solution was added dropwise to quench. After returning to room temperature, 0.5mL water was added dropwise. After filtration, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 330mg of crude compound 67 -2.

Step 3: Preparation of compound 67-3

330 mg of compound 67-2 was dissolved in 10 mL of anhydrous THF. Under nitrogen, 0 ^o C was added 60mg NaBH _4, reaction was brought to room temperature stirred for 0.5hrs. LCMS detected that the reaction was complete, and quenched by adding 5 mL of water dropwise under ice bath, returning to room temperature, concentrated in vacuo and spin-dried THF, the aqueous phase was extracted with DCM (5 mL×3), the combined organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 387 mg of yellow solid compound 67-3.

Step 4: Preparation of compound 67-4

200 mg of compound 67-3 and 99 mg of compound 22-3 were dissolved in 5 mL of dioxane, and 36 mg of Pd ₂ (dba) ₃ , 45 mg of Xantphos and 151 mg of N,N-diisopropylethylamine were added. Nitrogen thrice, under nitrogen, the reaction was raised to 70 ^o C the reaction was stirred for 2hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 160 mg of compound 67-4.

Step 5: Preparation of compound 67

50 mg of compound 67-4 was dissolved in 1.5 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.5 mL, methanol solution) was added, and the reaction was stirred at RT for 0.5 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (0.5 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (0.5 mL), dissolve the solid with DCM, dry and concentrate in vacuo , Purified by preparation plate to obtain 13.3 mg of white solid compound 67.

[M+H ⁺ ]=454.37.

¹ H NMR (500 MHz, DMSO- d ₆ ) δ 8.34 (s, 1H), 7.96 (d, J = 5.5 Hz, 1H), 7.31 (dd, J = 5.5, 8.0 Hz, 1H), 7.04-6.94 ( m, 2H), 6.73 (s, 2H), 6.08 (d, J = 5.5 Hz, 1H), 5.51 (t, J = 6.0 Hz, 1H), 4.52 (d, J = 5.5 Hz, 2H), 3.89- 3.78 (m, 3H), 3.20-3.10 (m, 2H), 3.05 (d, J = 16.0 Hz, 1H), 2.63 (d, J = 16.0 Hz, 1H), 1.90-1.85 (m, 1H), 1.82 -1.76 (m, 1H), 1.56-1.53 (m, 1H), 1.17-1.13 (m, 1H).

Example 68 Preparation of Compound 68:

Step 1: Preparation of compound 68-1

500 mg of compound 68-0 and 418 mg of compound M1-2 were dissolved in 10 mL of dioxane, 95 mg of Pd ₂ (dba) ₃ , 120 mg of Xantphos, and 900 mg of DIEA were added, and nitrogen was replaced three times. Raise the temperature to 100°C and react for 16hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 0.7 g of compound 68-1 as a pale yellow solid.

Step 2: Preparation of compound 68-2

0.7 g of compound 68-1 was dissolved in 3 mL of THF, 0.66 mg of tBuOK was added at room temperature, and the reaction was stirred at room temperature for 0.5 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, and the filter cake was washed with THF (5 mL×2) to obtain 380 mg of compound 68-2 as a gray solid.

Step 3: Preparation of compound 68-3

200 mg of compound M22-2 and 123 mg of compound 68-2 were dissolved in 4 mL of dioxane, 74 mg of Pd ₂ (dba) ₃ , 23 mg of Xantphos, and 104 mg of DIEA were added and replaced with nitrogen three times. The reaction solution was heated to 70°C and reacted for 3hrs. TLC detected that the reaction was complete, concentrated, and the residue was purified by preparative thin layer chromatography to obtain 101 mg of white compound 68-3.

Step 4: Preparation of compound 68

101 mg of compound 68-3 was dissolved in 1.5 mL of dioxane and 0.5 mL of MeOH, 2N HCl (632 μL, methanol solution) was added, the reaction was stirred at RT for 1 hrs, and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (4 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (4 mL), take the filter cake and vacuum dry to obtain 50.3 mg of beige solid compound 68.

[M+H ⁺ ]=450.25.

¹ H NMR (500 MHz, CDCl3) δ 8.36 (s, 1H), 8.01 (br, 1H), 7.34 (d, J = 6.5 Hz, 1H), 7.26-7.21 (m, 3H), 6.24 (d, J = 5.5 Hz, 1H), 5.02-5.00 (m, 1H), 4.70 (s, 2H), 4.01 (s, 1H), 3.64-3.59 (m, 2H), 3.23-3.14 (m, 2H), 3.10 ( d, J = 16.0 Hz, 1H), 2.89 (br, 3H), 2.74 (d, J = 15.5 Hz, 1H), 1.99-1.94 (m, 1H), 1.90-1.84 (m, 1H), 1.67-1.64 (m, 1H), 1.43-1.40 (m, 1H).

Example 69: Preparation of Compound 69:

Step 1: Preparation of compound 69-1

Under the protection of N ₂ , dissolve 2.23g PPh ₃ in 8mL dioxane, add 1.15g NCS, stir at room temperature for 0.5hrs, the reaction solution becomes thick white mud, add 0.7g compound M5, and heat to 100°C, Stir for 1hrs. TLC detected that the reaction was complete, cooled to room temperature, 7 mL of triethylamine, stirred for 15 mins, concentrated the reaction solution, and the residue was purified by column chromatography to obtain 260 mg of compound 69-1 as a brown oil.

Step 2: Preparation of compound 69-2

0.26g of compound 69-1, 342mg of compound M6 and 240mg of DIPEA were dissolved in 3mL of dioxane, and the reaction was stirred at 50°C for 2.5hrs. TLC detected that the reaction was complete, the reaction solution was concentrated, and the residue was purified by column chromatography to obtain 280 mg of compound 69-2 as a white solid.

Step 3: Preparation of compound 69-3

280 mg of compound 69-2 was dissolved in 3 mL of anhydrous DCM. Under nitrogen, -78 ^o C was slowly added dropwise 2.55 mL 1.0 M DIBAL-H in hexane solution, -78 ^o C the reaction was stirred for 0.5hrs. LCMS detected that the reaction was complete. The reaction was quenched by adding 0.1 mL of water and 0.1 mL of 2.5 M NaOH aqueous solution at low temperature. Return to room temperature and then adding 0.25 mL of water. After filtration, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 250 mg of crude compound 69. -3.

Step 3: Preparation of compound 69-4

250 mg of compound 69-3 was dissolved in 5 mL of anhydrous THF. 18mg NaBH _{4 was} added, and the reaction was stirred at room temperature for 0.5 hrs. LCMS detected that the reaction was complete, 5 mL of water was added dropwise to quench, then DCM (5 mL×2) was added for extraction, the combined organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 180 mg of yellow solid compound 69-4.

Step 4: Preparation of compound 69-5

180 mg of compound 69-4 and 90 mg of compound M1 were dissolved in 5 mL of dioxane, and 130 mg of Pd ₂ (dba) ₃ , 41 mg of Xantphos and 92 mg of N,N-diisopropylethylamine were added. Nitrogen thrice, under nitrogen, the reaction was raised to 70 ^o C the reaction was stirred for 3hrs. LCMS and TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 118 mg of compound 69-5.

Step 5: Preparation of compound 69

Under the protection of nitrogen, dissolve 118 mg of compound 69-5 in 1.5 mL of dioxane and 0.5 mL of MeOH, add 2N HCl (632 μL, methanol solution), stir at room temperature and react for 1 hrs. TLC detects that the reaction is complete and the reaction solution is decompressed concentrate. Add H ₂ O (4 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (4 mL), take the filter cake and vacuum dry to obtain 63.2 mg of beige solid compound 69.

[M+H+]=450.29

¹ H NMR (500 MHz, DMSO-d6) δ: 7.94 (d, J = 5.5 Hz, 1H), 7.36-7.30 (m, 1H), 7.20-7.15 (m, 3H), 6.70 (s, 2H), 5.98 (d, J = 5.5 Hz, 1H), 5.44 (t, J = 6.0 Hz, 1H), 4.48 (d, J = 5.5 Hz, 2H), 3.96-3.83 (m, 3H), 3.21-3.02 (m , 2H), 3.07 (d, J = 15.5 Hz, 1H), 2.64 (d, J = 15.5 Hz, 1H), 2.43 (s, 3H), 1.92-1.87 (m, 1H), 1.82-1.75 (m, 1H), 1.56-1.53 (m, 1H), 1.17-1.14 (m, 1H).

Example 70 Preparation of compound 70:

Step 1: Preparation of compound 70-1

Dissolve 5.0 g of compound M22-2 in 50 ml of methanol, add ammonia water (60 ml, 25%), and then react at RT for 2.5 hours. TLC detects that the reaction is complete, concentrates under reduced pressure (revolve most of the methanol), add MeOH (10ml) and stir for 5 minutes, filter, wash the filter cake with MeOH (3ml), dry the filter cake to obtain 4.75g of solid compound 70- 1.

Step 2: Preparation of compound 70-2

3.5 g of compound 70-1, 3.82 g of compound M6 and 5.04 g of DIPEA were dissolved in 40 mL of THF, and the reaction was stirred at 55°C for 12 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (40 ml), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 5.66 g of compound 70-2 as a white solid.

Step 3: Preparation of compound 70-3

100 mg of compound 70-2 and 45 mg of compound 22-3 were dissolved in 2 mL of dioxane, and 18 mg of Pd ₂ (dba) ₃ , 23 mg of Xantphos and 51 mg of N,N-diisopropylethylamine were added. Nitrogen thrice, under nitrogen, the reaction was raised to 100 ^o C the reaction was stirred for 3hrs. LCMS and TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 42 mg of compound 70-3.

Step 4: Preparation of compound 70

Under the protection of nitrogen, dissolve 42 mg of compound 70-3 in 0.5 mL of dioxane and 0.5 mL of MeOH, add 2N HCl (0.19 mL, methanol solution), stir and react at RT for 1 hrs, TLC detects that the reaction is complete, and the reaction solution is decompressed concentrate. Add H ₂ O (2 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO ₃ , solid precipitate, filter, wash the filter cake with H ₂ O (2 mL), take the filter cake and vacuum dry to obtain 17.8 mg of light yellow solid compound 70.

[M+H+]=449.32

¹ H NMR (500 MHz, DMSO- d ₆ ) δ: 8.34 (s, 1H), 8.00 (s, 1H), 7.96 (d, J = 4.5 Hz, 1H), 7.64 (s, 1H), 7.31 (d , J = 6.5 Hz, 1H), 7.20-7.13 (m, 3H), 6.70 (s, 2H), 6.08 (d, J = 4.5 Hz, 1H), 4.02-3.97 (m, 2H), 3.85 (s, 1H), 3.29-3.21(m, 2H), 3.07 (d, J = 15.5 Hz, 1H), 2.65 (d, J = 15.5 Hz, 1H), 1.83-1.69 (m, 2H), 1.53-1.50 (m , 1H), 1.15-1.12 (m, 1H).

[Comparative example]

Comparative example 1

According to the method described in EXAMPLE 48 in WO2018172984, the following Comparative Example 1 was prepared.

Comparative example 2

According to the method described in Example 243 in WO2019183367, the following Comparative Example 2 was prepared.

[Pharmacological test]

Example A: SHP2 allosteric inhibition enzyme activity assay

SHP2 is allosterically activated by the binding of a bis-tyrosinyl-phosphorylated peptide to its Src homology 2 (SH2) domain. This subsequent activation step results in the release of the auto-inhibition interface of SHP2, which in turn activates the SHP2 protein tyrosine phosphatase (PTP) and can be used for substrate recognition and reaction catalysis. In the rapid fluorometric format, the surrogate DiFMUP was used to monitor the catalytic activity of SHP2.

experiment procedure:

(1) Compound preparation:

The compound of the present invention (10mM stock solution) was diluted to an appropriate multiple with 100% DMSO, and the final test concentration of the compound of the present invention was 10μM, 3.3333μM, 1.1111μM, 0.3704μM, 0.1235μM, 0.0412μM, 0.0137μM, 0.0046μM, 0.0015μM , 0.00μM;

(2) Prepare working solution for enzyme reaction:

In a 96-well black polystyrene plate (flat bottom, low flange, non-binding surface) (Perki Elmer, Cat#6005270) at room temperature, use a final reaction volume of 50 μL and the following assay buffer conditions for SHP2 enzyme activity detection: 60mM HEPES, 75mM NaCl, 75mM KCl, 0.05% BRIJ-35, 1mM EDTA, 5mMDTT.

(3) Enzyme catalytic reaction and data monitoring:

Take the compound of the present invention and add it to the corresponding 96-well plate, and set the blank test well without adding the compound and enzyme and only adding buffer. Put SHP2 Activating Peptide (IRS1_pY1172(dPEG8)pY1222) on ice to melt, add 25μM to each well, then add 0.2ng SHP2 protein sample to the corresponding well plate, and incubate at room temperature for 1 hour. The replacement substrate DiFMUP (Invitrogen, Cat#D6567) was added to the reaction and reacted at room temperature for 2 hours. The fluorescence signal was monitored by a microplate reader (Envision, Perki Elmer) with excitation wavelength and emission wavelength of 340nm and 450nm, respectively.

(4) Data analysis:

Calculation formula: Inhibition rate%=[1-( Conversion_ _sample -Conversion_ _min )/( Conversion_ _max -Conversion_ _min )]×100% where: Conversion_sample is the conversion rate reading of the sample; Conversion_min is the average value of the blank control well, which means no enzyme activity The conversion rate reading of the well; Conversion_max is the mean value of the ratio of the positive control well, representing the conversion rate reading of the well without compound inhibition. Analysis using the GraphPad Prism software log (inhibitor) vs. response-Variable slope fitting dose-response curve, and calculating the activity of the compounds IC ₅₀ value pairs.

The IC ₅₀ data of some examples are shown in Table 4.

Table 4 Compound number Compound to SHP2 enzyme activity IC ₅₀ (nM) Compound 2 8.56 Compound 4 3.19 Compound 5 12.64 Compound 7 6.12 Compound 8 4.95 Compound 9 5.43 Compound 10 13.64 Compound 11 13.82 Compound 12 7.19 Compound 13 20.37 Compound 14 20.66 Compound 15 4.97 Compound 16 6.10 Compound 17 10.9 Compound 18 10.3 Compound 19 2.17 Compound 20 3.28 Compound 21 0.62 Compound 22 0.37 Compound 23 2.1 Compound 24 2.62 Compound 25 1.3 Compound 26 2.41 Compound 27 1.65 Compound 29 2.0 Compound 30 0.78 Compound 31 1.0 Compound 32 1.47 Compound 33 0.9 Compound 34 1.7 Compound 35 0.7 Compound 36 1.19 Compound 37 1.2 Compound 38 0.6 Compound 39 0.71 Compound 40 1.80 Compound 41 0.81 Compound 42 0.85 Compound 43 2.41 Compound 44 0.83 Compound 45 1.60 Compound 46 1.10 Compound 47 0.60 Compound 48 1.7 Compound 49 4.1 Compound 50 0.7 Compound 51 / Compound 52 / Compound 53 2 Compound 54 1 Compound 55 9.3 Compound 56 6.8 Compound 57 / Compound 58 0.72 Compound 59 2.15 Compound 60 7.19 Compound 61 19.75 Compound 62 1.4 Compound 63 4.3 Compound 64 2.0 Compound 65 2.1 Compound 66 3.1 Compound 67 2.0 Compound 68 1.8 Compound 69 4.7 Compound 70 2.3

Note: "/" means not tested.

The compound of the present invention has an allosteric inhibitory effect on SHP2 protein.

Example B: Cell Proliferation Test

In vitro cell assays were used to evaluate the effects of the compounds of the present invention on the proliferation of lung squamous cell carcinoma KYSE-520 cells and leukemia cells MV-4-11 cells. The detection method used in the experiment is the CELL TITER-GLO (CTG) luminescence method, which can detect the number of living cells by quantitatively measuring ATP. Because ATP participates in a variety of enzymatic reactions in organisms and is an indicator of living cell metabolism. Its content directly reflects the number and cell state of cells. During the experiment, CellTiter-Glo™ reagent was added to the cell culture medium to measure the luminescence value, luminescence value and The amount of ATP is directly proportional, and ATP is positively related to the number of live cells, so cell viability can be inspected by detecting ATP content.

experiment procedure:

(1) Cell plating:

Take the logarithmic phase of a bottle of KYSE-520 cells were counted after digestion of resuspended cells, seeded in a 96 well plate to adjust the cell density in seeded 1000 cells per well, plates disposed 37 ^o C, 5% CO ₂ in After culturing in an incubator for 24hrs, adding the compound of the present invention for treatment;

Take a bottle of MV-4-11 cells in the logarithmic growth phase, digest and resuspend the cells, count them, adjust the cell density and inoculate them into a 96-well plate, inoculate 4000 cells per well, and place the plate at 37 ^o C, 5% CO _2. After culturing in the incubator for 24 hours, add the compound of the present invention for treatment;

(2) Cell compound treatment:

Prepare appropriate amount of the compound of the present invention for cell treatment, and the final concentration of the compound from high to low is 1000nM, 333.3nM, 111.1nM, 37.04nM, 12.35nM, 4.115nM, 1.372nM, 0.4572nM, 0.1524nM, 0nM. Put it in a 37 ^o C, 5% CO ₂ incubator for 120 hrs. Only adding culture medium without adding cell wells was set as the zero adjustment group; the compound concentration of 0 nM group was the blank group.

(3) CTG test:

After cells are cultured for 120hrs, add 50μL of CellTiter-Glo® Luminescent Cell Viability Assay solution to each well, gently shake for 2mins, continue incubating for 10mins at room temperature, and read the detection value of each well on the multifunctional microplate reader.

(4) Data analysis: Calculate the inhibition rate based on the luminous value reading, Inhibition rate%=(1-(administration group value-zero adjustment group value)/(blank group value-zero adjustment group value)*100

GraphPad Prism the log (inhibitor) vs. response-Variable slope fitting dose-response curve and calculate IC ₅₀ of compounds to inhibit cell proliferation.

The experimental data is shown in Table 5.

table 5 Compound number Compound IC ₅₀ (nM) against KYSE-520 (EGFR amplification) cells Compound against MV-4-11 cells IC ₅₀ (nM) Compound 2 / 13.86 Compound 3 72.6 10.2 Compound 4 14.39 2.47 Compound 5 3.89 0.36 Compound 7 4.51 1.36 Compound 8 9.36 2.43 Compound 9 6.52 5.16 Compound 10 13.24 3.59 Compound 11 7.50 1.77 Compound 12 4.00 1.05 Compound 13 12.22 2.07 Compound 14 6.37 1.32 Compound 15 10.00 0.95 Compound 16 8.06 1.97 Compound 17 1.53 3.98 Compound 18 40.0 8.5 Compound 19 1.37 0.25 Compound 20 3.88 0.10 Compound 21 0.10 0.05 Compound 22 1.43 0.13 Compound 23 3.73 0.75 Compound 24 1.00 0.20 Compound 25 3.79 0.83 Compound 26 1.39 0.27 Compound 27 6.2 0.4 Compound 29 2.41 0.16 Compound 30 0.25 0.08 Compound 31 2.97 1.13 Compound 32 1.49 0.07 Compound 33 4.02 0.27 Compound 34 26.29 4.57 Compound 35 11.1 0.6 Compound 36 / / Compound 37 5.10 0.61 Compound 38 5.70 0.63 Compound 39 0.21 0.09 Compound 40 2.73 0.31 Compound 41 0.34 0.12 Compound 42 0.41 0.11 Compound 43 1.23 0.11 Compound 44 5.80 0.17 Compound 45 1.77 0.11 Compound 46 0.47 0.08 Compound 47 2.05 0.23 Compound 48 3.6 0.9 Compound 49 30.3 2.0 Compound 50 3.3 0.2 Compound 51 / / Compound 52 / / Compound 53 3.6 0.4 Compound 54 1.5 0.2 Compound 55 / 21.4 Compound 56 18.0 2.0 Compound 57 / / Compound 58 1.27 0.43 Compound 59 12.33 5.43 Compound 60 2.24 0.20 Compound 61 59.79 10.59 Compound 62 4.8 0.6 Compound 63 / 14.6 Compound 64 / 1.0 Compound 65 / 3.6 Compound 66 / 2.2 Compound 67 / 1.7 Compound 68 / 0.5 Compound 69 / 11.9 Compound 70 44.6 1.8

Note: "/" means not tested.

The compound of the present invention has a good inhibitory effect on the proliferation of KYSE-520 cells and the proliferation of MV-4-11 cells.

Example C: Inhibition test of hERG potassium ion channel

The whole cell patch clamp technique was used to detect the blocking effect of the test compound on the hERG channel. Cell culture

The HEK293 cell line stably expressing hERG potassium channels was cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg/mL G418 at a temperature of 37 ^o C and a carbon dioxide concentration of 5%.

Cell passage: Remove the old medium and wash once with PBS, then add 1 mL TrypLE™ Express solution, and incubate at 37°C for 0.5 minutes. When the cells detach from the bottom of the dish, add 5 mL of complete medium pre-warmed at 37°C. The cell suspension was gently pipetted to separate the aggregated cells. Transfer the cell suspension to a sterile centrifuge tube, and centrifuge at 1000 rpm for 5 minutes to collect the cells. To expand or maintain the culture, inoculate the cells in a 6 cm cell culture dish, each cell culture dish, the amount of cells inoculated is 2.5*105 cells (final volume: 5 mL).

In order to maintain the electrophysiological activity of the cells, the cell density must not exceed 80%.

For patch clamp detection, the cells were separated with TrypLE™ Express before the experiment. 3*103 cells were spread on a cover glass and cultured in a 24-well plate (final volume: 500 µL). After 18 hours, the experiment was performed.

Intracellular and extracellular fluid

Extracellular fluid: 140 mM NaCl, 3.5 mM KCl, 1 mM MgCl ₂ • 6H ₂ O, 2 mM CaCl ₂ , 10 mM D-glucose, 10 mM HEPES, 1.25 mM NaH ₂ PO ₄ , NaOH to adjust pH=7.4.

Intracellular fluid: 20 mM KCl, 115 mM K-Aspartic, 1 mM MgCl ₂ • 6H ₂ O, 5 mM EGTA, 10 mM HEPES, 2 mM Na ₂ -ATP, KOH to adjust pH=7.2. Compound formulation

Dilute the stock solution of the test compound with extracellular fluid to prepare a 10 µM working solution, or serially dilute to 0.3 µM, 1 µM, 3 µM, 10 µM, and 30 µM solutions.

The solubility of the test compound was visually inspected, and the test compound was completely dissolved without visible precipitation.

Cisapride (positive control)

The weighed 1.2 mg cisap must be made into a 10 mM stock solution using 243 µL DMSO.

Dilute the cisapride stock solution with DMSO successively in a 10-fold dilution from high to low to 10 µM dilution.

Dilute the 10 µM cisapride diluent with extracellular fluid to prepare a 10 nM working solution. Visually inspecting the solubility of cisapride, cisapride was completely dissolved without visible precipitation.

Experimental method reference:

The whole cell patch clamp voltage stimulation scheme for recording the whole cell hERG potassium current is as follows: when the whole cell seal is formed, the cell membrane voltage is clamped to -80 mV. The clamping voltage is depolarized from -80 mV to -50 mV for 0.5 seconds, then stepped to 30 mV for 2.5 seconds, and then quickly restored to -50 mV for 4 seconds to stimulate the tail current of the hERG channel. Collect data repeatedly every 10 seconds to observe the effect of drugs on hERG tail current. The leakage current was detected with a -50 mV stimulation for 0.5 seconds. The experimental data was collected by EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

The capillary glass tube is drawn into a recording electrode with a microelectrode drawing instrument. Operate the microelectrode manipulator under the inverted microscope to touch the recording electrode to the cell, and apply negative pressure suction to form a GΩ seal. After forming the GΩ seal, perform fast capacitance compensation, and then continue to give negative pressure to suck and break the cell membrane to form a whole-cell recording mode. Then perform slow capacitance compensation and record the film capacitance and series resistance. No leakage compensation is given.

When the hERG current recorded by the whole cell stabilizes, the drug is administered, and each drug concentration acts for 5 minutes (or the current stabilizes). Place the cell-covered cover glass in the recording bath in the inverted microscope. The test compound and the compound-free external fluid flow through the recording bath sequentially by gravity perfusion to act on the cells, and the vacuum pump is used to carry the liquid during recording. exchange. The current detected by each cell in the compound-free external fluid serves as its own control group. Test multiple cells independently and repeatedly. All electrophysiological experiments were performed at room temperature.

Data quality standards

The following criteria are used to determine whether the data is acceptable: (1) Series resistance ≤ 20 MΩ (2) Sealing resistance ≥ 1 GΩ (3) Peak tail current peak ≥ 400 pA (4) The starting tail current peak value is greater than the starting current peak value (5) There is no obvious spontaneous attenuation of tail current (the spontaneous attenuation is less than 5% within 5 minutes) (6) There is no obvious leakage current under the membrane potential of -80 mV (leakage current ≤ 100 pA)

)，然後計算每一個藥物濃度對應的抑制率

。Data analysis first standardizes the current after each drug concentration and the blank control current (

), and then calculate the inhibition rate corresponding to each drug concentration

.

See Table 6 for the hERG test results of the example compounds.

Table 6 Numbering Inhibition rate @10µM Comparative example 1 95.12% Comparative example 2 ＞95.6% Compound 3 86.7% Compound 5 60.8% Compound 8 97.36% Compound 9 93.35% Compound 16 87.99% Compound 18 42.63% Compound 21 82.3% Compound 22 43.5% Compound 24 79.9% Compound 26 ＞90% Compound 30 92.2% Compound 33 ＞90% Compound 39 83.7% Compound 40 98.1% Compound 41 85.6% Compound 42 87.6% Compound 43 56.6% Compound 44 80.2% Compound 45 78.0% Compound 46 87.8% Compound 58 69.4% Compound 59 57.0% Compound 62 ＞90% Compound 64 5.02% Compound 65 62.1% Compound 66 37.1% Compound 67 29.5% Compound 68 54.4% Compound 69 26.3% Compound 70 9.12%

We unexpectedly found that while the compound of the present invention generally has good inhibitory activity on SHP2, when one of the following two conditions is met, namely 1), the position of R ₁ on the pyrazine ring in the general formula I is a hydroxyl group. A methyl group and the positions of A ₁ and A ₃ are both N, such as compound 22, compounds 64 to 69; 2). In the general formula I, the R ₁ position on the pyrazine ring is an amide group and the A ₃ position is N, such as compound 5 Compared with other technical solutions, compound 18, compound 58 or compound 59 can significantly improve the hERG inhibition problem and is expected to reduce cardiotoxicity.

Example D: Plasma protein binding assay

Follow the steps below to determine plasma protein binding.

1) Preparation of 100 mM sodium phosphate and 150 mM NaCl buffer (PBS)

An alkaline solution was prepared by dissolving 14.2 g/L Na ₂ HPO ₄ and 8.77 g/L NaCl in deionized water, which can be stored at 4°C for up to 7 days. An acidic solution can be prepared by dissolving 12.0 g/L NaH ₂ PO ₄ and 8.77 g/L NaCl in an acidic solution, which can be stored at 4°C for 7 days. The alkaline solution was titrated with an acidic solution to pH 7.4 and stored at 4°C for 7 days. Check on the day of the experiment and adjust if the pH exceeds the specification 7.4±0.1.

2) Preparation of plasma

The frozen plasma is immediately thawed at room temperature.

The plasma was centrifuged at 3,220 g for 10 minutes to remove clots, and the supernatant was collected in a new test tube. Check and record the pH of the plasma.

Note: a). Only use plasma that has been frozen and thawed not more than twice. b). Use only plasma in the range of pH 7 to pH 8.

3) Preparation of working solution

Working solutions of the test compound and the reference compound ketoconazole were prepared with DMSO at a concentration of 400 μM. Then remove 4 μL of the working solution and mix it with 796 μL of human, dog, monkey, rat or mouse plasma to obtain a mixed solution with a concentration of 2 μM (0.5% DMSO). Vortex the plasma sample thoroughly.

4) Quickly balance the dialysis steps

Add 200 μL of plasma sample on the red circle side and 400 μL of dialysis buffer (PBS) on the opposite side for dialysis. The determination is made in triplicate. The dialysis plate was sealed and incubated in a 37°C incubator at 150 rpm for 4 hours. At the end of the incubation, remove the seal and transfer 50 μL of samples from the buffer chamber and the plasma chamber to the wells of the 96-well plate.

5) Sample analysis steps

Add 50 μL of blank plasma to each buffer sample, and add an equal volume of PBS to the collected plasma sample. 300 μL room temperature quenching solution (acetonitrile containing internal standard (IS, 10 ng/mL labetalol and 10 ng/mL glibenclamide)) was added to precipitate the protein. The sample in the plate was vortexed for 5 minutes and centrifuged at 3220 g for 30 minutes at 4°C. Then use 100 μL or 200 μL of water to transfer 100 μL of the supernatant to a new 96-well plate for LC-MS/MS analysis (depending on the LC-MS signal response and peak shape).

Calculate the percentage of test compound and control compound binding as follows:% free = (peak area ratio _{buffer chamber} /peak area ratio _{plasma chamber} ) *100% binding = 100-% free% recovery = (peak area ratio _{buffer chamber} + Peak area ratio ( _{plasma chamber} ) / Peak area ratio _{total sample} *100 Peak area ratio _{buffer chamber} indicates the concentration of the free part Peak area ratio _{plasma chamber} indicates the concentration of the free and bound parts Peak area ratio _{Total sample} indicates the concentration of the sample started before incubation

Table 7 shows the plasma protein binding results of the control compound and the test compound in different species.

Table 7 Compound Average% binding rate people Rat Mouse (naked) dog monkey Comparative example 1 98.07 98.95 98.87 98.63 96.77 Comparative example 2 99.7 99.5 99.4 99.4 99.4 5 91.7 96 97.4 90.6 91.4 12 95.5 97.7 97.6 95.5 97.3 18 93 98 98 87.5 95.9 20 93.6 98.2 92.5 85 91.8 twenty one 97.1 97.4 97.5 96 97.1 twenty two 76.1 89.4 85.2 73.7 84.2 twenty four 96.9 98.3 97.2 96.3 95.3 30 97.5 97.8 95.2 96.3 93.5 31 93.1 95.4 93.9 91.5 91.3 39 91.9 92.1 91.6 91.3 90 43 99 96.3 95.8 94.1 98.8 44 96.6 98.1 96.8 96.7 96 47 85.8 90.2 89 82.6 79.7 48 84.7 87.4 79.3 71.7 90.9 50 95.5 96.2 94.6 96.1 95.1 53 93.9 93.4 94 92.1 89.8 58 87.1 92.3 91.9 93.2 90 59 95.2 97.1 96.8 95.5 92.4 60 96.8 96.5 97.4 93.2 96.9 62 96.5 97.8 97.8 97.3 97.7 63 97.6 98.5 98.8 98.8 97.9 64 92.2 93.7 91.7 81 91.3 65 83.7 92.6 77 63.2 71.2 66 97.2 97.7 96.4 94.3 95.4 67 93.4 93 91.5 82.2 90 68 96.2 94.3 88 85.7 93 69 91.4 90.4 89.7 83.4 87.9 70 69.4 93.3 88.3 67.1 75.5

Usually, only the unbound part has a biological effect or is metabolized. Therefore, the degree of binding to plasma proteins will significantly affect the pharmacokinetics and pharmacodynamic properties of the drug.

As shown in Table 7, the comparative example reflects a high degree of binding to plasma proteins, so the efficacy of the drug may be reduced. Unexpectedly, the exemplary compound of the present invention has a lower degree of plasma protein binding compared to the comparative example. It is predicted that the present invention has excellent pharmacokinetics and pharmacodynamic properties on the human body.

Although the present invention has been fully described through its embodiments, it is worth noting that various changes and modifications are obvious to those skilled in the art. Such changes and modifications should be included in the scope of the attached patent application of the present invention.

no.

no.

no.

Claims (47)

A compound represented by formula I, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,

Formula I wherein, R ₁ optionally selected from _{amino, -C (O) -R a,} -C≡N, hydroxy, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 Alkoxy, C _1-8 alkoxy containing substituents; R _{a is} optionally selected from amino, -NH-OH, C _1-3 alkyl; R _{2 is} optionally selected from hydrogen, C _1-4 alkyl Or C _1-4 alkyl containing substituents; R _{3 is} arbitrarily selected from hydrogen, halogen, amino, -C(O)NH ₂ , -C≡N, hydroxy, C _1-8 alkyl, substituents containing C _1-8 alkyl, C _1-8 alkoxy or substituted C _1-8 alkoxy; R _{4 is} optionally selected from hydrogen, halogen, amino, amide, -C≡N, carboxy, Hydroxy, hydroxymethyl, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _2-8 alkenyl , C _2-8 alkenyl group having a substituent, C _2-8 alkynyl or C _2-8 alkynyl group-containing substituent group; a ₁ is selected from optionally CR ₅ or N; a ₂ is selected from CR ₆ or optionally N; A _{3 is} arbitrarily selected from CR ₇ or N; A _{4 is} arbitrarily selected from CR ₈ or N; U is arbitrarily selected from C(R ₉ ) ₂ , O or NR ₁₀ ; wherein, R ₅ , R ₆ , R ₇ , R ₈ , R ₉ or R _{10 are} independently selected from hydrogen, hydroxy, halogen, amino, substituted amino, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy group, a substituted C _1-8 alkoxy group, C _2-8 alkenyl group, a substituted group containing C _2-8 alkenyl, C _2-8 alkynyl group, a substituted group containing C _2-8 alkynyl Or C _5-6 heterocyclic group; or, R ₅ and R ₆ and the carbon atom to which they are attached together form a 5- to 6-membered aryl group or a 5- to 6-membered heterocyclic group; L is selected from S; ring A is optionally selected From C _6-10 aryl or C _5-10 heteroaryl, the C _5-10 heteroaryl contains one or two N or S heteroatoms; Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, Amino, substituted amino, sulfonyl, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _3-8 cycloalkyl or substituted C _3-8 cycloalkyl; n is 0, 1, 2, 3 or 4. The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R _{1 is} optionally selected from amino, -C(O)NH ₂ , -C≡N, C _1-3 alkyl, substituted C _1-3 alkyl or C _1-3 alkoxy. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R _{1 is} arbitrarily selected from Amino, -C(O)NH ₂ , -C≡N, hydroxyl substituted methyl,

,

,

,

,

or

. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R ₂ Is hydrogen. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R ₃ optionally selected from hydrogen, halo, C _1-3 alkyl or a C _1-3 alkyl group containing substituent. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R ₃ Arbitrarily selected from hydrogen, chlorine or methyl. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R ₄ Arbitrarily selected from hydrogen, halogen, C _1-8 alkyl group containing substituents, and C _1-8 alkoxy group. The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R ₄ Arbitrarily selected from hydrogen, F, Cl, -CHF ₂ , CF ₃ or -O-CH ₃ . The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₁ It is CR ₅ or N, wherein R _{5 is} selected from hydrogen, halogen or halogen substituted C _1-3 alkyl. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₁ Is CR ₅ or N, wherein R _{5 is} selected from hydrogen, Cl or trifluoromethyl. The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₂ It is CR ₆ or N, wherein R _{6 is} selected from hydrogen, hydroxyl, halogen, amino or C _1-8 alkoxy. The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₂ It is CR ₆ or N, wherein R _{6 is} selected from hydrogen, OH, F, Cl, amino or -O-CH ₃ . The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₃ Is CR ₇ or N, where R ₇ is hydrogen or halogen. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₄ Is CR ₈ or N, wherein R _{8 is} selected from hydrogen, halogen, amino, substituted amino, C _1-3 alkoxy or C _5-6 heterocyclic group. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₄ Is CR ₈ or N, wherein R _{8 is} selected from hydrogen, F, Cl, amino, -NHCH ₃ , -N(CH ₃ ) ₂ ,

,

Or methoxy. The compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein U is CH ₂ or O. The compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein ring A Arbitrarily selected from phenyl or C _5-6 heteroaryl, the C _5-6 heteroaryl contains one or two N or S heteroatoms. The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein Rx is any Ground is selected from hydrogen, hydroxy, halogen, cyano, C _1-3 alkyl, halogen-substituted C _1-3 alkyl or C _1-3 alkoxy. The compound according to any one of claims 1 to 18, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein Rx is any Ground is selected from hydrogen, OH, F, Cl, Br, -CN, trifluoromethyl or methoxy. The compound according to any one of claims 1 to 19, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein

Selected from

,

,

,

,

,

,

,

,

,

,

or

. The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is represented by formula II :

Formula II wherein, R ₃ is optionally selected from hydrogen, halogen, _{amino, -C (O) NH 2,} -C≡N, hydroxy, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 alkoxy or C _1-8 alkoxy containing substituents; A _{1 is} optionally selected from CR ₅ or N; U is optionally selected from C(R ₉ ) ₂ , O or NR ₁₀ ; wherein, R _5. R ₉ or R _{10 are} independently selected from hydrogen, hydroxy, halogen, amino, substituted amino, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy containing substituted C _1-8 alkoxy group, C _2-8 alkenyl group, a substituted group containing C _2-8 alkenyl, C _2-8 alkynyl group or a substituted group containing C _2-8 alkynyl or C _5-6 heterocycle; or, L is selected from S; R _{x is} arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C _1-8 alkyl, substituted C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _3-8 cycloalkyl or substituted C _3-8 cycloalkyl; n is 0, 1, 2, 3, or 4. The compound according to claim 21, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R _{3 is} optionally selected from hydrogen or methyl. The compound according to claim 21 or 22, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₁ is CR ₅ , Where R _{5 is} selected from F or Cl. The compound according to claim 21 or 22, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₁ is N. The compound according to any one of claims 21 to 24, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein U is O or CH ₂ . The compound according to any one of claims 21 to 25, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R _x Arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C _1-3 alkyl, halogen-substituted C _1-3 alkyl, or C _1-3 alkoxy. The compound according to any one of claims 21 to 26, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R _x Arbitrarily selected from hydrogen, OH, CN, Cl, F, trifluoromethyl or methoxy. The compound according to any one of claims 21 to 27, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein n is 0, 1, or 2. The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is represented by formula III :

Optionally the formula III R ₃ is selected from hydrogen, halogen, _{amino, -C (O) NH 2,} -C≡N, hydroxy, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _{l- 8} Alkoxy or C _1-8 alkoxy containing substituents; R _{4 is} optionally selected from hydrogen, halogen, amino, amide, -C≡N, carboxyl, hydroxyl, C _1-8 alkyl, containing Substituent C _1-8 alkyl, C _1-8 alkoxy, substituted C _1-8 alkoxy, C _2-8 alkenyl, substituted C _2-8 alkenyl, C _{2 -8} alkynyl or C _2-8 alkynyl containing substituents; A _{1 is} optionally selected from CR ₅ or N; A _{2 is} optionally selected from CR ₆ or N; A _{3 is} optionally selected from CR ₇ or N; U Arbitrarily selected from C(R ₉ ) ₂ , O or NR ₁₀ ; wherein R ₅ , R ₆ , R ₇ , R ₈ , R ₉ or R _{10 are} independently selected from hydrogen, hydroxyl, halogen, amino, and substituents amino, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 alkoxy, group containing C _1-8 alkoxy substituent, C _2-8 alkenyl group, a substituted C _2-8 alkenyl, C _2-8 alkynyl, C _2-8 alkynyl or C _5-6 heterocyclic group containing substituents; L is selected from S; ring A is optionally selected from C _6-10 Aryl or C _5-10 heteroaryl, the C _5-10 heteroaryl contains one or two N or S heteroatoms; Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, and substituents amino, sulfo acyl, C _1-8 alkyl, C _1-8 alkyl-containing substituent group, a C _1-8 alkoxy group, a substituted C _1-8 alkoxy group, C _3-8 cycloalkyl Alkyl group or C _3-8 cycloalkyl group containing substituents; n is 0, 1, 2, 3 or 4. The compound according to claim 29, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R _{3 is} selected from hydrogen. The compound according to claim 29 or 30, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein R _{4 is} arbitrarily selected from Hydrogen or Cl. The compound according to any one of claims 29 to 31, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₁ Is CR ₅ or N, wherein R _{5 is} selected from hydrogen, Cl or trifluoromethyl. The compound according to any one of claims 29 to 32, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₂ Is CR ₆ or N, where R _{6 is} selected from Cl, amino,

, Or -O-CH ₃ . The compound according to any one of claims 29 to 33, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein A ₃ Is CR ₇ or N, where R ₇ is hydrogen. The compound according to any one of claims 29 to 34, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein U is CH ₂ . The compound according to any one of claims 29 to 35, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein ring A Arbitrarily selected from phenyl or C _5-6 heteroaryl, the C _5-6 heteroaryl contains one or two N or S heteroatoms. The compound according to any one of claims 29 to 36, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein Rx is any Ground is selected from hydrogen, Cl, Br, trifluoromethyl or methoxy. The compound according to any one of claims 29 to 37, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein

Selected from

,

,

,

,

,

,

,

. The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is: 2)( S )-(3-(1-amino-1,3-dihydrospiro[inden-2,4'-piperidine] -1'-yl)-6-((2,3-dichlorophenyl)sulfur (Substituted)-5-methylpyrazin-2-yl)methanol; 3)( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-Yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazine-2-carbonitrile; 4)( S )-3-(1-amino-1,3-di Hydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2,3-dichlorophenyl)sulfanyl)pyrazine-2-methanamide; 5)( S ) -3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine] -1'-yl)-6-((2-amino-3-chloropyridin-4-yl) Sulfuryl)pyrazine-2-carboxamide; 7)( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)- 6-((3-Chloro-2-methoxypyridin-4-yl)thio)pyrazine-2-carboxamide; 8)( S )-3-(1-amino-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl)pyridin-3-yl)thio)pyrazine-2-carboxamide; 9 )( S )-3-(1-amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-( (Trifluoromethyl)pyridin-3-yl)thio)pyrazine-2-carboxamide; 10)( S )-6-((2-amino-3-chloropyridin-4-yl)thio)- 3-(1-Amino-4-bromo-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide; 11)( S )- 6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-methoxy-1,3-dihydrospiro[indene-2,4'-呱(Pyridin)-1'-yl)pyrazine-2-carboxamide; 12)( S )-6-((2-amino-3-chloropyridin-4-yl)thio)-3-(1-amino -6-Methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide; 13)( S )-6-(( 2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro[indene-2,4'-piperidine] -1'-yl)pyrazine-2-carboxamide; 14)( S )-6-((2-amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6 -(Trifluoromethyl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide; 15)( S )-6-( (2-Amino-3-chloropyridine (Pyridin-4-yl)thio)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)pyrazine-2- Formamide; 16) ( S )-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((6-amino- 3-chloropyridin-2-yl)thio)pyrazine-2-carboxamide; 17) ( S )-3-(5-amino-2-methoxy-5,7-dihydrospiro[cyclopentyl Diene [ b ]pyridine-6,4'-pyridine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazine-2-methanamide ; 18) ( S )-3-(4-Amino-2-chloro-4,6-dihydrospiro[cyclopenta[ d ]thiazole-5,4'-piperidine]-1'-yl) -6-((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide; 19) (S)-(3-(1-amino-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)-6-((2-fluoro-3-methoxyphenyl)thio)pyrazin-2-yl)methanol; 20) ( S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-methoxypyrimidin-4-yl )Thio)pyrazin-2-yl)methanol; 21) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl )-6-((2-Amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)methanol; 22) (S)-(3-(1-amino-1,3-di Hydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 23) (S) -1-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine-4 -Yl)thio)pyrazin-2-yl)ethan-1-one; 24) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine ]-1'-yl)-6-((3-chloro-2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazine-2- Yl)methanol; 25) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-( (Trifluoromethyl)pyridin-4-yl)thio)pyrazin-2-yl)methanol; 26) (S)-(6-((2-amino-3-chloropyridin-4-yl)thio) -3-(1-amino-6-(trifluoromethyl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol; 27) 1-(3-((S)-1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -chlorine Pyridin-4-yl)thio)pyrazin-2-yl)ethan-1-ol; 28) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-Pepidine)-1'-yl)-6-((5-chloro-2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazine-2-yl)methanol; 29) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(( 2-(Dimethylamino)-3-fluoropyridin-4-yl)thio)pyrazin-2-yl)methanol; 30) (S)-(3-(1-amino-1,3-dihydro) Spiro[indene-2,4'-pyridine]-1'-yl)-6-((3-fluoro-2-(methylamino)pyridin-4-yl)thio)pyrazin-2-yl)methanol ; 31) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(difluoromethyl Yl)pyridin-3-yl)thio)pyrazin-2-yl)methyl; 32) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'- Pyridin]-1'-yl)-6-((3-chloro-2-(dimethylamino)pyridin-4-yl)sulfanyl)pyrazin-2-yl)methyl; 33) (S) -(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-fluoro-2-methoxyphenyl) Sulfuryl)pyrazin-2-yl)methanol; 34) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl) -6-((3-chloro-5-fluoro-2-methoxyphenyl)thio)pyrazin-2-yl)methanol; 35) (S)-(3-(1-amino-1,3 -Dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(quinolin-4-ylthio)pyrazin-2-yl)methanol; 36) (S)-( 3-(1-Amino-1,3-dihydrospiro[inden-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl)pyridin-3-yl) Thio)pyrazin-2-yl)methanol; 37) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl) -6-((2,3-Dichlorophenyl)thio)pyrazin-2-yl)methanol; 38) (S)-(3-(1-amino-1,3-dihydrospiro[indene- 2,4'-Pepidine]-1'-yl)-6-((2-(trifluoromethyl)pyridin-3-yl)thio)pyrazin-2-yl)methanol; 39) (S) -(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-fluoropyridin-4-yl )Thio)pyrazin-2-yl)methyl; 40) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'- Yl)-6-((3-chloro-2-methoxypyridin-4-yl) Thio)pyrazin-2-yl)methanol; 41) (S)-(6-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-fluoro -1,3-Dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol; 42) (S)-(6-((2-amino-3 -Chloropyridin-4-yl)thio)-3-(1-amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyridine Azin-2-yl)methanol; 43) (S)-(6-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-5,6-dimethyl -1,3-Dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol; 44) (R)-(6-((2-amino-3 -Chloropyridin-4-yl)thio)-3-(3-amino-3H-spiro[benzofuran-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl ; 45) (S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl) -1,3-Dihydrospiro[indene-2,4'-piperidine]-4-carbonitrile; 46) (S)-(6-((2-amino-3-chloropyridin-4-yl)sulfanyl )-3-(1-Amino-6-chloro-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)pyrazin-2-yl)methanol; 47) (S )-(3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-(pyridin-4-ylthio)pyrazine-2 -Base) methanol; 48) (S)-(6-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(5-amino-5,7-dihydrospiro[cyclopentyl Eno[b]pyridine-6,4'-pyridine]-1'-yl)pyrazin-2-yl)methyl; 49) (S)-(6-((2-amino-3-chloropyridine) -4-yl)thio)-3-(1-amino-4-(trifluoromethyl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyridine (Azin-2-yl)methanol; 50) (S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl) Pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-pyridine]-6-ol; 51) (S)-4-((5-(1-amino-1, 3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyrazin-2-yl)thio)pyrimidin-2-ol; 52) (S )-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)-7-fluoro- 1,3-Dihydrospiro[indene-2,4'-piperidine]-4-ol; 53) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4 '-Pyridine]-1'-yl)- 6-((2,3-Dihydro-[1,4]dioxa[2,3-b]pyridin-8-yl)thio)pyrazin-2-yl)methanol; 54) ( S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3 -Dihydrospiro[indene-2,4'-piperidine]-6-nitrile; 55) (S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl )Thio)-3-(hydroxymethyl)pyrazin-2-yl)-4-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-7-nitrile; 56) ( S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-7-(trifluoromethyl)-1,3-dihydrospiro(indene -2,4'-Pepidine]-1'-yl)pyrazin-2-yl)methyl; 57) (S)-1-amino-1'-(5-((2-amino-3-chloro Pyridin-4-yl)thio)-3-(hydroxymethyl)pyrazin-2-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-7-ol; 58) (S)-3-(1-Amino-1,3-dihydrospiro[inden-2,4'-piperidine]-1'-yl)-6-((2-amino-3-fluoropyridine-4 -Yl)thio)pyrazine-2-carboxamide; 59) (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'- Yl)-6-((3-chloro-2-((1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazine-2-carboxamide; 60 ) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((2-amino-3-chloropyridine- 4-yl)thio)-N-hydroxypyrazine-2-carboxamide; 61) 1-(3-((S)-1-amino-1,3-dihydrospiro[indene-2,4'-Piperidine)-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)-2,2,2-trifluoroethane-1-Alcohol; 62) (S)-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(methoxymethyll)pyrazin-2-yl )-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine; 63) (S)-2-(3-(1-amino-1,3-dihydrospiro[indene -2,4'-Pepidine]-1'-yl)-6-((2-amino-3-chloropyridin-4-yl)thio)pyrazin-2-yl)propan-2-ol; 64 ) (S)-(3-(1-Amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2- Aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 65) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine ]-1'-yl)-6-(pyrimidin-4-ylthio)pyridine Azin-2-yl)methanol; 66) (S)-(3-(1-amino-5-chloro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl) -6-((2-Aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 67) (S)-(3-(1-amino-5-fluoro-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol; 68) (S)- (3-(1-amino-1,3-dihydrospiro[inden-2,4'-piperidine]-1'-yl)-6-((2-(methylamino)pyrimidin-4-yl) Thio)pyrazin-2-yl)methanol; 69) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl) -6-((2-Aminopyrimidin-4-yl)thio)-5-methylpiperazine-2-yl)methanol; or 70) (S)-3-(1-amino-1,3-di Hydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidin-4-yl)thio)pyrazine-2-carboxamide. A pharmaceutical composition comprising a therapeutically effective amount of at least one compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, or non-covalent Complex or prodrug; and at least one pharmaceutically acceptable excipient. The pharmaceutical composition according to claim 40, wherein the compound, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, and a pharmaceutically acceptable salt The mass percentage of acceptable excipients is 0.0001:1-10. A compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, or claim 40 Or the use of the pharmaceutical composition described in 41 in the preparation of medicines. The application according to claim 42, wherein the medicament is used to treat, prevent, delay or prevent cancer, cancer metastasis, cardiovascular disease, immune disease, fibrosis or eye disease. The use according to claim 42, wherein the drug is used as an SHP2 inhibitor. A compound according to any one of claims 1 to 39, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, or claim 40 Or the use of the pharmaceutical composition described in 41 in the preparation of a medicine for treating diseases mediated by SHP2. The application according to claim 45, wherein the disease is cancer. The use according to claim 43 or 46, wherein the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute Myeloid leukemia, breast cancer, esophageal tumor, lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma, pancreatic cancer, or a combination thereof.