WO2020156243A1

WO2020156243A1 - Shp2 inhibitor and application thereof

Info

Publication number: WO2020156243A1
Application number: PCT/CN2020/072774
Authority: WO
Inventors: 吴颢; 吴文茂; 陈忠研; 李玲; 林远望; 吴云飞; 朱林强; 袁丁; 王丰; 张展; 冯东杰; 赵新涛; 余军; 束庆玉; 程见洪; 韩晗; 郭晶; 兰宏; 王家炳; 丁列明
Original assignee: 贝达药业股份有限公司
Priority date: 2019-01-31
Filing date: 2020-01-17
Publication date: 2020-08-06
Also published as: TW202043212A; CN113365988A; CN113365988B

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

Technical field

The present invention relates to a series of compounds as inhibitors of Src homology region 2 (Src homology region 2-containing protein tyrosine phosphatase 2, SHP2), and preparation methods and pharmaceutical compositions 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.

Background technique

Src homology region 2 protein tyrosine phosphatase 2 (Src homology region 2-containing protein tyrosine phosphatase 2, SHP2) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene, and PTPN11 is the first The discovered proto-oncogene encoding tyrosine kinase (Chan R J 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 rich Proline (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 growth factor receptor is activated, its tyrosine residues undergo autophosphorylation, which is the combination of Grb2 and SHP2 (adapter protein containing SH2 domain) phosphotyrosine Area SH2 provides stop points. The binding of Grb2 to phosphorylated growth factor receptors results in the accumulation of SOS protein in the cell membrane. SOS, as a guanine nucleotide exchange factor (GEF), can catalyze the conversion of membrane-bound protein Ras from inactive Ras-GDP to active Ras-GTP. Ras-GTP further connects with downstream signaling systems to activate Ser/Thr kinase Raf1, etc., and then activate ERK under the action of the regulatory kinase MEK. After activation of ERK, it 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 Features.

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 Noonan 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. 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; Bentiers-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, there is no marketed product for this target at home and abroad. Therefore, the development of small molecule drugs that can target and inhibit the activity of SHP2 and provide patients with safer and more effective SHP2 inhibitors have important research significance.

Summary of the invention

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 a general structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,

among them,

Optionally R ₁ is 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;

Optionally R ₂ is selected from hydrogen, C _1-4 alkyl group or a substituted C _1-4 alkyl;

Optionally 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 _1-8 alkoxy Oxy or C _1-8 alkoxy containing substituents;

Optionally R ₄ is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, hydroxymethyl, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 alkoxy, C substituent group containing _1-8 alkoxy, C _2-8 alkenyl, C _2-8 alkenyl-containing substituent, C _2-8 alkynyl group or a substituted group containing C _2-8 alkynyl base;

A _{1 is} arbitrarily selected from CR ₅ or N;

A _{2 is} arbitrarily selected from CR ₆ or 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, hydroxyl, halogen, amino, substituted amino, C _1-8 alkyl, and substituted 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, C _2-8 alkynyl or C _5-6 heterocyclic group containing substituents; 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 a C _6-10 aryl group or a C _5-10 heteroaryl group, the C _5-10 heteroaryl group 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 containing substituted C _1-8 alkoxy group, C _3-8 cycloalkyl or an unsubstituted C _3-8 cycloalkyl group;

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.

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

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.

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.

In some embodiments, the formula I

Selected from

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,

among them,

A _{1 is} arbitrarily selected from CR ₅ or N;

U is arbitrarily 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, C substituent group containing _1-8 alkoxy, C _2-8 alkenyl, C _2-8 alkenyl-containing substituent, C _2-8 alkynyl group or a substituted group containing C _2-8 alkynyl Group or C _5-6 heterocycle; or,

L is selected from S;

R _{x 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 group, a substituted group containing C _1-8 alkoxy, C _3-8 cycloalkyl or C _3-8 cycloalkyl group having substituent;

n is 0, 1, 2, 3, or 4.

In some embodiments, R ₃ in Formula II is optionally 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.

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 R ₄ is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, C _1-8 alkyl, substituted group containing C _1-8 alkyl, C _1-8 alkoxy, A substituted C _1-8 alkoxy group, a C _2-8 alkenyl group, a substituted C _2-8 alkenyl group, a C _2-8 alkynyl group or a substituted C _2-8 alkynyl group;

A _{1 is} arbitrarily selected from CR ₅ or N;

A _{2 is} arbitrarily selected from CR ₆ or N;

A _{3 is} arbitrarily selected from CR ₇ 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, hydroxyl, halogen, amino, substituted amino, C _1-8 alkyl, and substituted 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, C _2-8 alkynyl or C _5-6 heterocyclic group containing substituents;

L is selected from S;

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.

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.

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'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)-5-methylpyrazin-2-yl)methanol;

3) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine) -4-yl)thio)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-chloropyridine) -4-yl)thio)pyrazine-2-carboxamide;

7) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methoxy (Pyridin-4-yl)thio)pyrazine-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-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'-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'-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-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2, 4'-Piperidine]-1'-yl)pyrazine-2-carboxamide;

16)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((6-amino-3-chloropyridine -2-yl)thio)pyrazine-2-carboxamide;

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-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-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-fluoro-3-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;

20)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-methoxypyrimidine- 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-chloro Pyridin-4-yl)thio)pyrazin-2-yl)methanol;

22)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-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 -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-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-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-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-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-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'-piperidine]-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-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;

34)(S)-(3-(1-amino-1,3-dihydrospiro[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-ylsulfanyl )Pyrazin-2-yl)methanol;

36)(S)-(3-(1-amino-1,3-dihydrospiro[indene-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'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)pyrazin-2-yl)methanol;

38)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-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-fluoro Pyridin-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-methyl Oxypyridin-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)pyrazin-2-yl)methanol;

43)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-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'-Piperidine]-1'-yl)pyrazin-2-yl)methanol;

47)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(pyridin-4-ylsulfanyl) Pyrazin-2-yl)methanol;

48)(S)-(6-((2-amino-3-chloropyridin-4-yl)thio)-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-dihydro Spiro[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)pyridine (Azin-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'-piperidine]-1'-yl)-6-((2,3-dihydro- [1,4]Dioxo[2,3-b]pyridin-8-yl)thio)pyrazin-2-yl)methanol;

54)(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-nitrile;

55)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-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-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl;

57)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-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-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'-piperidine]-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-ol;

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'-piperidine]-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) Pyrazin-2-yl)methanol;

66)(S)-(3-(1-Amino-5-chloro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;

67)(S)-(3-(1-amino-5-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;

68)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(methylamino) Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;

69)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-4- Yl)thio)-5-methylpiperazin-2-yl)methanol; or

70)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((2-aminopyrimidin-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 preferred 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 tumor , 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 a subject.

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 a subject.

Preferably, in the above method, the treatment target is a human.

Unless otherwise specified, 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 includes but is 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. Similar, "C _1-8 alkyl""C_1-8" means comprising 7 or 8 carbon atoms, a straight chain, branched chain or cyclic Formally arranged 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 6 to 10 membered monocyclic or bicyclic aromatic group. Preferably it is phenyl and naphthyl. 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 groups include: 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, ring The other atoms on the above 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, other atoms in the ring Is 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, triazole, indolyl, benzofuranyl, benzothiazolyl, benzimidazolyl, benzopyrazolyl, benzene And similar groups such as triazolyl, carbazolyl, quinolinyl, isoquinolinyl, purinyl and the like.

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, the result is a heteroaryl group instead of an aryl group.

"Cycloalkyl" refers to a cyclic group that is saturated or unsaturated but not aromatic. Depending on the specific level of saturation, the terms "cycloalkyl", "cycloalkenyl" or "cycloalkynyl" are used respectively. Representative cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclohexane, or cyclohexene and similar groups. Specifically, the cycloalkyl group may be a C _3-10 cycloalkyl group, such as a C _3-6 cycloalkyl group.

"Heterocyclic group" refers to a saturated or unsaturated but not aromatic cyclic group, and one or more of the carbon atoms (and the connected hydrogen atoms) 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 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 to form a di-, poly- or spiro-ring, it will still be defined as a heterocycloalkyl.

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). Preferred halogen refers to fluorine, chlorine and bromine.

"Halo" refers to a fluoro, chloro, bromo or iodo group. The preferred halo groups refer 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, hydroxy, oxo, carbonyl, cyano, -C (O) NH _2, alkyl, alkoxy, aryl, cycloalkyl, heterocyclyl , Heteroaryl. In some embodiments, the substituent includes, but is not limited to, halogen, amino, hydroxy, cyano, methyl, -CH ₂ OH, -C(O)NH ₂ , -OCH ₃ , trifluoromethyl.

Whenever the term "alkyl" or "aryl" or its prefixes appear in the name of a substituent (such as aralkyl, or dialkylamino), it shall be the same 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 prefix stem) 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 well known for use in animals, especially those that can be used in humans.

The term "composition" in the present invention includes a product containing a specific quantity of a specific component, and also includes any product directly or indirectly obtained from a specific quantity 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, A dose that produces a certain effect in the treatment of symptoms, indications or discomfort. The specific "effective therapeutic dose" may vary according to 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. Whenever 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 the pharmaceutically acceptable salt includes a pharmaceutically acceptable acid/anion or base/cation salt. 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, cyclohexylamine sulfonic acid, salicylic acid, saccharic acid or trifluoroacetic acid. Pharmaceutically acceptable base/cation salts, including, but not limited to, aluminum salt, calcium salt, chloroprocaine salt, choline, diethanolamine salt, ethylenediamine salt, lithium salt, magnesium salt, potassium salt, 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 a desired compound in vivo. 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 in vivo after administration to the subject. disease. The conventional methods for selecting and preparing suitable prodrug derivatives have been recorded in books such as "Design of Prodrugs" (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, salts of ammonium, calcium, magnesium, potassium, and sodium 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, including 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 Accessories. 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 composition for parenteral (including subcutaneous administration, intramuscular injection, intravenous administration) 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 through 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 with 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, Microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, 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.

Tablets containing the compound or pharmaceutical composition of the present invention can be prepared by, optionally, mixing, compression or molding with one or more auxiliary components or adjuvants. 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 wetted 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 the 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 glycerol, liquid polyethylene glycol, and their mixtures in oil, dispersion systems can also be prepared. Furthermore, 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 preferred. 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, 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. These preparations can be prepared by using the compound represented by the formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, through conventional processing methods. As an example, an emulsion or ointment is prepared by adding a hydrophilic material and water (the total amount of the two is about 5 wt% to 50 wt% of the compound) to prepare a cream or ointment with the desired 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. Suppositories in which the mixture forms a unit dose are the most preferred dosage form. Suitable excipients include cocoa butter and other materials commonly used in the art. Suppositories can be conveniently prepared. First, the pharmaceutical composition is mixed with softened or melted excipients, then cooled and molded.

In addition to the above-mentioned carrier components, the above-mentioned pharmaceutical preparations may also include, as appropriate, one or more additional adjuvant components, such as diluents, buffers, flavoring agents, binders, surfactants, and additives. Thickeners, lubricants, preservatives (including antioxidants), etc. Further, other adjuvants may also include penetration enhancers that regulate the isotonic pressure of the drug and 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.

detailed description

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

The following abbreviations are used in the examples:

ACE-Cl: 1-chloroethyl chloroformate;

(BOC) ₂ O: di-tert-butyl dicarbonate;

BOP: Benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate;

DBU: 1,8-diazabicycloundec-7-ene;

DCE: 1,2-dichloroethane;

DCM: dichloromethane;

DIPEA or DIEA: N,N-diisopropylethylamine;

DMAc: N,N-dimethylacetamide;

DMF: N,N-dimethylformamide;

DMSO: dimethyl sulfoxide;

EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride;

EtOAc or EA: ethyl acetate;

EtOH: ethanol;

EtONa: sodium ethoxide;

h, hr or hrs: hour;

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'-bisdiphenylphosphinoferrocene palladium dichloride;

Pd ₂ (dba) ₃ : Tris(dibenzylideneacetone)dipalladium;

Pd(OAc) ₂ : Palladium(II) acetate;

PE: petroleum ether;

PPA: polyphosphoric acid;

rt, r.t. 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 85°C and stirred for 12 hrs. 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

20.00g of compound M1-3 was dissolved in 200mL THF, EtONa (35mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT 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 15 g of compound M1 as a solid.

Preparation of intermediate compound M2:

Step 1: Preparation of compound M2-2

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

Step 2: Preparation of compound M2-3

Dissolve compound 1.10g M2-2 in 15mL dioxane, add 491mg compound M1-2, 46mg Xantphos, 35mg Pd(OAc) ₂ and 1.05g DIEA, replace the mixture with nitrogen 3 times, heat to 90°C for 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°C, slowly add EtONa's EtOH solution (2mL, 20%) to the above solution, stir and react at -30°C for 30mins, and slowly rise to room temperature Stir the reaction for 2hrs. TLC detected the completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was added 20mL DCM to be slurried for 30mins, filtered, the filter cake was washed with DCM (5mL×3), and the filter cake was vacuum-dried to obtain 990mg of brown solid M2.

Preparation of intermediate compound M3:

Step 1: Preparation of compound M3-2

Dissolve 3.00g of compound M3-1 and 1.60g of compound M1-2 in 30mL of dioxane, add 243mg of Pd ₂ (dba) ₃ , 384g of Xantphos and 3.40g of DIPEA, replace with nitrogen three times, raise the reaction solution to 110℃ and stir. Reaction 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 of compound M3-2 was dissolved in 50mL THF, EtONa (7.4mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 2hrs. TLC detected the completion of the reaction, concentrated under reduced pressure, added 50mL DCM and stirred for 30mins, the reaction solution was filtered, the filter cake was washed with DCM (10mL×2), and drained to obtain 3.60g 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 85°C and stirred for 12 hrs. 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

266 mg of compound M4-3 was dissolved in 5 mL of THF, EtONa (0.47 mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 3 hrs. 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 the protection of nitrogen, 10.60g of compound M5-2 was dissolved in 26mL of ethanol, the reaction solution was cooled to 0°C, and 25.00g of M5-1 was added dropwise. After dropping, the ice bath RT was removed for 2hrs, and then heated at 90°C for overnight reaction. 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

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

Preparation of intermediate compound M6:

Step 1: Preparation of compound M6-3

Under the protection of nitrogen, dissolve 25.00g of compound M6-1 in 200mL of DMF, reduce the temperature to 0℃, add 22.70g NaH in batches, keep at 0℃ for 1hr, and then slowly add 54.96g compound M6-2 dropwise to the reaction solution After dropping, react at 0°C for 1 hr, and then heat to 60°C to continue the reaction 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, and concentrated under reduced pressure. The residue was passed through the column Analyze and purify to obtain 29.00g brown oil M6-3.

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 12 hrs. 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 sulfuric acid It was dried with sodium and concentrated 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 the protection of nitrogen, 48.00g of compound M6-5 was dissolved in 500mL of anhydrous THF, the temperature was lowered to -20°C, 6.73g of NaHB ₄ was slowly added, and then the temperature was raised to RT and stirred for 2hrs. After the reaction, the reaction solution was cooled to 0°C, quenched with 300 mL of water, extracted with EtOAc (300 mL×3), combined the organic phases, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was passed through a 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°C, quenched with 100 mL saturated aqueous NaHCO ₃ , extracted with EtOAc:THF=3:1 (100 mL×3), combined the organic phases, washed with saturated brine, 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(500MHz,DMSO-d ₆ )δ7.26-7.21(m,4H), 5.80(d,J=10.5Hz,1H), 4.43(d,J=10.5Hz,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°C and 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

1.50 g of compound M7-3 was dissolved in 15 mL of toluene, 1.1 mL of PBr _{3 was} added dropwise, the reaction solution was raised to 105° C. and stirred for 12 hrs. 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

Dissolve 450 mg of compound M7-5 in 6 mL DMF, add 271 mg of NaH in batches at 0°C, stir and react at 60°C for 1 hr under nitrogen protection, then add 1.20 g of compound M4-4, and stir at 60°C for 1 hr. TLC detected the completion of the reaction. The reaction was quenched by adding 30 mL of water, and extracted with EtOAc (25 mL×2) and water (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography. 160 mg of compound M7-6.

Step 4: Preparation of compound M7-7

160 mg of compound M7-6 was dissolved in 2 mL of DCE at 0°C, 155 mg of ACE-Cl was added dropwise, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, 4mL MeOH was added, and the reaction was raised to 80°C and 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

Dissolve 4.00g of compound M8-1 in 50mL of anhydrous THF, replace with nitrogen three times, cool to -78°C, slowly add LDA solution in THF (11.70mL, 2.0M) dropwise, and then react at -78°C for 1 hr to convert compound M8 -2 in THF (10 mL) was slowly added dropwise to the above reaction solution, reacted at -78°C for 30 mins, and slowly raised to room temperature for 2 hrs. TLC detected the completion of the reaction. The reaction was quenched with 30 mL saturated NH ₄ Cl solution, 50 mL water was added, and EtOAc was added for extraction (60 mL×3). The organic phases were combined, washed with 50 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was purified by column chromatography to obtain 4.30 g of colorless oil M8-3.

Step 2: Preparation of compound M8-4

4.30g of compound M8-3 was dissolved in THF/MeOH (40mL/40mL), NaOH aqueous solution (20mL, 2.4N) was added, and the reaction was heated to 80°C for 18hrs. TLC detected the completion of the reaction, the reaction solution was cooled to room temperature, concentrated under reduced pressure 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 vacuum dried 3.40 g of compound M8-4 was obtained as a white solid.

Step 3: Preparation of compound M8-5

3.40g of compound M8-4 was dissolved in 40mL PPA, and the temperature was raised to 120°C to react for 2hrs. TLC detected that the reaction was complete. The reaction solution was slowly added dropwise to 200 mL 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 M9-2

10.00 g of compound M9-1 was dissolved in 100 mL of MeOH, 2.0 mL of concentrated sulfuric acid was added, and the reaction was heated at 70° C. for 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, and concentrate under reduced pressure. Purification by column chromatography yielded 9.23 g of white solid, compound M9-2.

Step 2: Preparation of compound M9-3

Dissolve 9.23 g of compound M9-2 in 150 mL of MeOH, cool to 0° C. in an ice bath, add 6.97 g of NaBH _{4 in} batches, and naturally increase the temperature to RT for 5 hrs. After the completion of the reaction, 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 Colored liquid, compound M9-3.

Step 3: Preparation of compound M9-4

Under nitrogen protection, dissolve 3.00g of compound M9-3 in 50mL of dichloromethane, cool the reaction solution to -15°C, add 2.81mL of NEt ₃ , and then add 1.04mL of MsCl solution dropwise. After the addition is complete, the temperature is raised to 0 React at ℃ for 1hr. After the reaction was completed, 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 the protection of nitrogen, dissolve 3.39 g of compound M9-5 in 20 mL of anhydrous THF, lower the temperature to -50°C, and add 1.71 g of LDA solution dropwise. After dripping, keep the temperature at -50°C and react for 1 hour. 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 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

Dissolve 4.61g of compound M9-6 in 8mL of water and 40mL of MeOH, and add 2.07g of NaOH. The temperature was raised to 65°C and stirred 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 a column Chromatographic 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℃, add NaH (60%, 0.42g) in batches, and then keep at -15℃ for 1hr. Then, the temperature was lowered to -60°C, n-butyllithium (1.6M, 7.8mL) was added dropwise, and the reaction was kept warm for 1hr. After the reaction was completed, 50 mL of water was added, extracted with EtOAc (50 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 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 the protection of nitrogen, 2.83 g of compound M10-2 was dissolved in 50 mL of anhydrous THF, the temperature was lowered to -78° C., and a THF/Hex solution of LDA (2M, 6 mL) was added dropwise. Incubate at -78°C for 1 hr, then slowly drop 1.69 g of compound M10-1 in THF (3 mL) into the reaction solution, and react at -78°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 1.44 g of a pale yellow oil compound M10- 3.

Step 2: Preparation of compound M10-4

Under the protection of nitrogen, 900 mg of compound M10-3 was dissolved in 50 mL of anhydrous THF, and the temperature was lowered to -78°C. A THF/Hex solution of LDA (2M, 3 mL) was added dropwise. The reaction was incubated at -78°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, the intermediate compounds in Table 1 are prepared by a method similar to the aforementioned intermediate 6-10 M11-M15.

Table 1

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. Replace with nitrogen three times and stir at 80°C for 12 hrs under nitrogen protection. 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

228 mg of compound 2-2 was dissolved in 3 mL of POCl ₃ , and 38 mg of compound N,N-dimethylbenzamide was added, and the reaction was raised to 110° C. and stirred for 1.5 hrs. 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 Press and concentrate to obtain 195 mg of compound 2-3.

Step 3: Preparation of compound 2-4

Dissolve 195mg of compound 2-3 and 236mg of compound M6 in 3mL NMP, and add 268mg DIEA. The reaction was raised to 80°C and 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

165 mg of compound 2-4 was dissolved in 2.5 mL THF at 0° C., 24 mg LiAlH _{4 was} added in batches, and stirred for 10 mins. TLC detected that the reaction was complete, and 50 μL of water, NaOH (15%, 50 μL), 150 μL of water and 1 g of anhydrous sodium sulfate were sequentially added at 0° C., and stirred for 10 mins. 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 aqueous NaHCO ₃ to adjust the pH to 7-8, extracts with DCM/MeOH = 10/1 (15 mL×3), combines the organic phases, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was purified by column chromatography to obtain 47.9 mg of compound 2.

[M+H ⁺ ]=501.13.

¹ H NMR (500MHz, DMSO-d ₆ ): δ7.48 (d, J=8.5Hz, 1H), 7.32-7.30 (m, 1H), 7.25 (t, J=8.5Hz, 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=16.0Hz,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 for 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

Dissolve 142 mg of compound 3-2, 62.3 mg of compound M1 and 55 mg of KI in dioxane and react in microwave at 120°C for 1 hr. LC-MS detected that the reaction was complete, 10 mL of water and 10 mL of EtOAc were added for extraction three times, the organic layers were combined, 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 reacted 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 NaHCO ₃ aqueous solution, add 10 mL of water after the solid is precipitated, and extract with EtOAc (10 mL×3). Combine the organic phases and dry with anhydrous sodium sulfate. After pressure 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 (500MHz, CDCl3): δ8.34 (s, 1H), 7.79 (d, J = 5.5Hz, 1H), 7.35-7.33 (m, 1H), 7.25 (m, 3H), 6.17 (d, J = 5.5Hz, 1H), 4.56-4.50 (m, 2H), 4.03 (s, 1H), 3.51-3.42 (m, 2H), 3.12 (d, J = 15.5Hz, 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-bis Preparation of chlorophenyl)thio)pyrazine-2-carboxamide

Step 1: Preparation of compound 4-2

14.96 mg of KI and 186.77 mg of K ₂ CO ₃ were added to the DMA (3 mL) solution of 200 mg of compound 4-1 and 104.86 mg of compound 2-1, and stirred at 120°C for 2 hrs. 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 by Pre-TLC (PE:EtOAc=3:1) After purification, 233 mg of compound 4-2 was obtained as a yellow solid with a yield of 88.18%.

[M+H ⁺ ]:586.23.

Step 2: Preparation of compound 4-3

NaOH (2.5M, 132.97 μL) was added dropwise to a mixed solution of 200 mg of compound 4-2 in dioxane (1.56 mL) and water (1.56 mL), and stirred at 100° C. for 12 hrs. The reaction solution was adjusted to pH 6-7 with 1N HCl, extracted with EtOAc (10mL×3), and 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°C, a methanol solution of hydrochloric acid (2M, 0.5 mL) was added dropwise to 95 mg of compound 4-3 in dioxane (3 mL), and the mixture was stirred at 25°C for 1 hr. 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), combined the organic phases, washed with saturated brine, and dried over anhydrous sodium sulfate After concentration, 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 (500MHz, CDCl ₃ ): δ8.20 (s, 1H), 7.35-7.33 (dd, J = 8.0, 1.5Hz, 1H), 7.32 (s, 1H), 7.25-7.19 (m, 4H) ,7.11(t,J=8.0Hz,1H),6.99(dd,J=8.0,1.5Hz,1H),5.40(s,1H),4.06-3.93(m,3H),3.39-3.29(m,2H ), 3.11(d,J=16.0Hz,1H),2.74(d,J=16.0Hz,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

Step 1: Preparation of compound 5-1

Dissolve 121 mg of compound 3-3 in 1.2 mL of 1,4-dioxane and 1.2 mL of water, and add 0.12 mL of 2.5M NaOH aqueous solution. The reaction solution was heated to 100°C for 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 the reaction was conducted at RT for 30 mins. 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 (500MHz, DMSO-d ₆ ): δ8.34(s,1H), 8.04(s,1H), 7.69-7.67(m,2H), 7.32(d,J=6.5Hz,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=15.5Hz,1H),2.66(d,J=15.5Hz,1H),1.83-1.70(m,2H),1.53-1.51(m,1H),1.16-1.14 (m,1H).

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

Table 2

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

¹ H NMR (500MHz, DMSO-d ₆ ): δ8.39(s,1H), 8.06(s,1H), 7.92(d,J=5.5Hz,1H), 7.69(s,1H), 7.31-7.30 (m,1H),7.19-7.16(m,3H),6.39(d,J=5.5Hz,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=15.4Hz,1H), 2.64(d,J=15.4Hz,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 (500MHz, DMSO-d ₆ ): δ8.55 (d, J = 4.5Hz, 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=6.5Hz,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=15.0Hz,1H),2.63(d,J=14.5Hz,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 (500MHz, CDCl ₃ ): δ8.55 (dd, J = 4.5, 1.0 Hz, 1H), 8.22 (s, 1H), 7.67 (d, J = 7.5 Hz, 1H), 7.39 (dd, J ＝8.2,4.5Hz,1H),7.12(br,1H),7.10(d,J＝8.3Hz,1H), 6.89(d,J＝2.5Hz,1H), 6.76(dd,J＝8.2,2.5Hz ,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=15.0Hz,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)

¹ H NMR (500MHz, DMSO-d ₆ ): δ8.36 (s, 1H), 8.04 (s, 1H), 7.69 (m, 2H), 7.38-7.13 (m, 3H), 6.38 (s, 2H) ,5.89(d,J=5.0Hz,1H),4.03-3.97(m,2H),3.94(s,1H),3.32-3.24(m,2H),3.04(d,J=17.0Hz,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)

¹ H NMR (500MHz, DMSO-d ₆ ): δ8.35(s,1H), 8.05(s,1H), 7.69(m,2H), 7.19(t,J=7.5Hz,1H), 6.95(d ,J=7.5Hz,1H),6.82(d,J=7.5Hz,1H),6.38(s,2H),5.88(d,J=5.0Hz,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(500MHz, CDCl ₃ ): δ8.28(s,1H), 7.75(d,J=5.0Hz,1H), 7.12(d,J=8.5Hz,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=15.0Hz,1H), 2.68(d,J=15.0Hz,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(500MHz, CDCl ₃ ): δ8.29(s,1H), 7.74(d,J=5.4Hz,1H), 7.60(s,1H), 7.49(d,J=7.8Hz,1H), 7.32(d,J=7.8Hz,1H),6.07(d,J=5.4Hz,1H),4.16-4.04(m,2H),4.03(s,1H),3.43-3.29(m,2H),3.18 (d,J=16.0Hz,1H),2.79(d,J=16.0Hz,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(500MHz,DMSO-d ₆ )δ8.51(s,1H), 8.38(s,1H), 8.03(s,1H), 7.78(d,J=5.4Hz,1H), 7.71-7.66( m,1H), 7.36(d,J=1.9Hz,1H),7.33-7.27(m,1H),7.21-7.13(m,3H),6.15(d,J=5.4Hz,1H), 6.09(d ,J=1.9Hz,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=15.6Hz, 1H), 2.64(d,J=15.6Hz,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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 85°C and stirred for 12 hrs. 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 was obtained.

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. LCMS detects that the reaction is complete, concentrates under reduced pressure, adds 5 mL of DCM and sonicates for 5 mins, then decanted the supernatant after standing, adds 5 mL of MTBE and sonicates for 5 minutes, after standing, decanted the supernatant, 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

Add 1.90g of NaH (content 60%) to 25mL of THF in batches to raise the temperature to 60°C under nitrogen protection, add dropwise a THF (40mL) solution of 5.00g of compound M16-2 and 3.46g of compound M16-1, and stir the reaction 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. 4.45g 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℃ and stir Reaction 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

5.38g of compound M16-4 was dissolved in 50mL THF, EtONa (9mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 2hrs. TLC detected the completion of the reaction, 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

700 mg of compound M17-SM and 2.5 mL of NH(CH ₃ ) ₂ aqueous solution were dissolved in 2.5 mL of dioxane, replaced with nitrogen three times, and the reaction was raised to 90° C. under the protection of nitrogen and stirred for 2 hours. TLC detects that the reaction is complete, 10 mL of water and 10 mL of ethyl acetate are added for extraction, the organic phase is separated, washed with 10 mL of saturated sodium chloride solution, and dried with 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 of dioxane, and 243 mg of Pd ₂ (dba) ₃ , 308 mg of Xantphos and 688 mg of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 5 hours. 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

584 mg of compound M17-2 was dissolved in 6 mL of THF, and an ethanol solution (4 mL) of EtONa (185 mg) and CH ₃ ONa (183 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 500 mg of crude compound M17-3, which could be used directly in the next reaction.

Preparation of intermediate compound M18-3:

Step 1: Preparation of compound M18-1

700 mg of compound M17-SM and 2.5 mL of methylamine aqueous solution were dissolved in 2.5 mL of dioxane, and replaced with nitrogen three times. Under the protection of nitrogen, the reaction was raised to 90° C. and stirred for 2 hr. TLC detects that the reaction is complete, 10 mL of water and 10 mL of ethyl acetate are added for extraction, the organic phase is separated, washed with 10 mL of saturated sodium chloride solution, and dried with 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 of Pd ₂ (dba) ₃ , 334 mg of Xantphos and 747 mg of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 5 hours. 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

646 mg of compound M18-2 was dissolved in 7 mL of THF, and an ethanol solution (3 mL) of EtONa (180 mg) and CH ₃ ONa (243 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 500 mg of crude compound M18-3, which can be directly used in the next reaction.

Preparation of intermediate compound M19-2:

Step 1: Preparation of compound M19-1

1 g of compound M19-SM and 578 mg of compound M1-2 were dissolved in 10 mL of dioxane, and 220 mg of Pd ₂ (dba) ₃ , 278 mg of Xantphos and 1.24 g of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 5 hours. 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 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 an ethanol solution (4 mL) of EtONa (268 mg) and CH ₃ ONa (251 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 700 mg of crude compound M19-2, which can be used directly in the next reaction.

Preparation of intermediate compound M20-3:

Step 1: Preparation of compound M20-1

1 g of compound M20-SM and 3 mL of NH(CH ₃ ) ₂ aqueous solution were dissolved in 3 mL of dioxane, replaced with nitrogen three times, and the reaction was raised to 90° C. under the protection of nitrogen and stirred for 2 hours. TLC detects the completion of the reaction, adds 10 mL of water and 10 mL of ethyl acetate for extraction, separates the organic phase, washes with 10 mL of saturated sodium chloride solution, and dried with 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 4 hours. 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 the ethanol solution (3 mL) of EtONa (120 mg) and CH ₃ ONa (228 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 900 mg of crude compound M20-3, which can be directly used in the next reaction.

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 25% ammonia solution and 20 mL dioxane, and the tube was sealed at 70° C. and the reaction was stirred for 48 hours. 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 3.67 g, Compound M21-1.

Step 2: Preparation of compound M21-2

2.92 g of compound M21-1 and 1.47 g of compound M1-2 were dissolved in 45 mL of dioxane, and 354 mg of Pd ₂ (dba) ₃ , 225 mg of Xantphos and 3.17 g of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 80°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 2.68g of compound M21-2 in 40mL THF, add dropwise EtONa (1.19g) ethanol (12mL) solution at a temperature of -30°C, and then naturally increase RT to react for 3hr. TLC detected that the reaction was complete, 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 used directly in the next reaction.

Preparation of intermediates M22-2 and M23-1:

Step 1: Preparation of compound M22-1

Dissolve 1.29 g of compound M22-SM and 1.60 g of compound M6 in 20 mL of tetrahydrofuran, add 1.12 g DIEA dropwise, and stir and react at 60°C for 12 hours. TLC detected the completion of the reaction, concentrated under reduced pressure, added 50 mL of water, and 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 protection, 1.00 g of compound M22-1 was dissolved in 30 mL of anhydrous dichloromethane, the temperature was reduced to -78°C, and DIBAL-H (1M, 9.6 mL) in n-hexane was added dropwise. The reaction was stirred at -78°C for 1 hour. Then slowly increase the temperature to -40°C and continue the reaction for 2 hours. TLC detected that the reaction of the raw materials was complete. At 0° C., 0.4 mL of water was slowly added dropwise, and then sodium hydroxide aqueous solution (15%, 0.4 mL) was added dropwise, and then 1 mL of water was added. 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

Dissolve 1.00 g of compound M24-1 in 10 mL of anhydrous acetonitrile, add 1.45 g of potassium carbonate, and stir and react at 85°C for 2 hours. TLC detected that the reaction was complete, the solvent was removed under reduced pressure, 50 mL of water was added, and then extracted with EA (50 mL×2), and the organic layers were combined, dried over sodium sulfate and desolventized, and purified by column chromatography to obtain 710 mg of compound M24-2.

Step 2: Preparation of compound M24-3

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 were dissolved in 10 mL of dioxane, replaced with nitrogen three times, and reacted at 85°C for 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and then 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 taken and dried under vacuum to obtain 60 mg of brown solid M24-4.

Preparation of intermediate compound M25-5:

Step 1: Preparation of compound M25-2

At 0°C, an aqueous solution (25 mL) of sodium hydroxide (546 mg) was added dropwise to an aqueous solution in which 1.00 g of compound M25-1, 2.60 g of elemental iodine and 1.70 g of potassium iodide were dissolved. The reaction was carried out at room temperature for 2 hours, and the reaction was completed as detected by TLC. Adjust the pH to neutral with saturated ammonium chloride solution. Then use sodium thiosulfate solution to remove 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

1.45g of compound M25-2, 1.13g of methyl iodide and 1.10 potassium carbonate were dissolved in DMF (20mL), reacted at room temperature for 3h, 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. Purified by column chromatography to obtain 1.19 g of compound M25-2.

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, replace with nitrogen three times, and react at 85°C for 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

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 were dissolved in 10 mL of dioxane, and after nitrogen replacement three times, the reaction was carried out at 85° C. for 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 nitrogen protection, 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 minutes, 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 sodium thiosulfate aqueous solution (100mL, 0.4mol/L), then add sodium hydroxide aqueous solution (70mL, 40%), and then extract with chloroform (150mL×2), and combine the organic layers After drying with sodium sulfate, the solution was removed and recrystallized with dichloromethane/n-hexane to obtain 8.77g of compound M27-3.

Step 2: Preparation of compound M27-5

8.77 g of compound M27-3 was dissolved in 300 mL of anhydrous tetrahydrofuran, the temperature was lowered to -30° C., a n-hexane solution of n-butyl lithium (2.5 M, 16 mL) was added dropwise, and the mixture was kept warm and stirred for 1 hour. Subsequently, 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°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

11.06g of compound M27-5 was dissolved in a mixed solution of water (40mL) and dichloromethane (200mL), and then 3.17g of pyridine, 12.83g of M27-6 and 0.86g of tetrabutylammonium bromide were sequentially added. Stir at room temperature for 24 hours. The reaction solution was quenched with 100 mL of saturated brine, extracted with DCM (100 mL×3), 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° 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 1.30g of compound M27-8 and 1.56g of (R)-(+)-tert-butylsulfinamide in a mixed solution of tetraethyl titanate (10mL) and anhydrous tetrahydrofuran (2mL), and replace with nitrogen 3 times Then, it was reacted at 90°C for 8 hours. The heating was stopped, the reaction solution was poured into 200 mL of ice water, 150 mL of ethyl acetate was added, and the mixture was stirred 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

Dissolve 1.30 g of compound M27-9 in 25 mL of anhydrous tetrahydrofuran, add 344 mg of sodium borohydride in batches at -20°C, slowly warm to room temperature, and then stir at room temperature for 3 hours. At 0°C, 100 mL of water was added, and then extracted with EtOAc (80 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 0.94 g of 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 was reduced to 0°C, and then saturated aqueous sodium bicarbonate solution was added to adjust the pH to weakly alkaline. It was extracted with a mixed solution of EtOAc:THF=1:1 (60 mL×4), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 692 mg of compound M27-11 was obtained.

Example 19 Preparation of compound 19:

Step 1: Preparation of compound 19-2

1.00 g of compound 19-1 was dissolved in 10 mL of anhydrous acetonitrile, 1.45 g of potassium carbonate was added, and the reaction was stirred at 85°C for 2 hours. TLC detected the completion of the reaction, 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 720 mg of compound 19-2.

Step 2: Preparation of compound 19-3

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 were dissolved in 10 mL of dioxane, replaced with nitrogen three times, and reacted at 85°C for 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 that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was added to 5mL DCM for 30min to be slurried, filtered, the filter cake was washed with DCM (5mL×3), and 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 it with nitrogen three times, and heat to 100°C for 3 hours. TLC tracks the reaction. 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 over sodium sulfate and desolventized, and purified by column chromatography to obtain 28 mg of compound 19-5.

Step 5: Preparation of compound 19

28 mg of compound 19-5 was dissolved in 1 mL of dioxane, and methanol solution of hydrochloric acid (2N, 0.4 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, 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 in vacuum. 15.7 mg of compound 19 was obtained as a pale yellow solid.

[M+H ⁺ ]=467.29

¹ H NMR(500MHz,DMSO-d ₆ ,):δ8.01(s,1H),7.31(d,J=6.6Hz,1H),7.22-7.08(m,5H),6.91(t,J=7.0 Hz, 1H), 5.35 (t, J = 5.0 Hz, 1H), 4.48 (d, J = 5.2 Hz, 2H), 3.85 (s, 3H), 3.68-3.64 (m, 2H), 3.33-3.30 (m ,1H),3.10-3.00(m,3H),2.64-2.59(m,1H),1.91-1.86(m,1H),1.80-1.76(m,1H),1.51(d,J=12.8Hz,1H ), 1.13-1.10 (m, 1H).

Preparation of compound 20:

Step 1: Preparation of compound 20-2

500 mg of compound 20-1 and 318 mg of compound M1-2 were dissolved in 6 mL of dioxane, 48 mg of Pd ₂ (dba) ₃ , 76 mg of Xantphos, and 684 mg of DIPEA were added, and nitrogen was replaced three times. The reaction solution was heated to 90° C. and stirred for 5 hrs. 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

569 mg of compound 20-2 was dissolved in 7 mL of THF, EtONa (1.3 mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 3 hrs. 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and 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 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 (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(500MHz,DMSO-d6)δ8.36(s,1H),8.32(d,J=5.3Hz,1H),7.38-7.33(m,1H),7.24-7.17(m,3H),6.82 (d,J=5.3Hz,1H), 5.50(d,J=6.3Hz,1H), 4.53(d,J=4.6Hz,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=15.7Hz,1H), 2.71(d,J=15.7Hz,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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 80°C and stirred for 12 hrs. 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°C, 11 mg LiAlH _{4 was} added in batches, and the reaction was stirred at 0°C for 0.5 hrs. 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. 12 mg of 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(500MHz,DMSO-d ₆ )δ8.31(s,1H), 7.67(d,J=5.4Hz,1H), 7.31(d,J=6.8Hz,1H), 7.23-7.11(m, 3H), 6.36(s, 2H), 5.91(d,J=5.3Hz,1H), 5.48(t,J=5.9Hz,1H), 4.52(d,J=5.9Hz,2H),3.94-3.83( m,3H),3.24-3.11(m,2H),3.06(d,J=15.5Hz,1H), 2.63(d,J=15.3Hz,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 extract the aqueous phase, combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, the residue is purified by column chromatography to obtain a class 0.605 g of compound 22-2 as a white solid.

Step 2: Preparation of compound 22-3

0.605g compound 22-2 was dissolved in 5mL THF, EtONa (5.75mL, 20% EtOH solution) was added dropwise at RT, and the reaction was stirred at RT for 3hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 15 mL of n-hexane and stirred for 30 mins, the reaction solution was filtered, and the filter cake was washed 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. to react for 3 hours. 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(500MHz,DMSO-d ₆ )δ8.33(s,1H),7.96(d,J=5.0Hz,1H), 7.31(d,J=7.0Hz,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=16.0Hz,1H), 2.63(d,J=15.5Hz,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. TLC detects the completion of the reaction. Adjust the reaction solution to pH=7 with 1N HCl solution, add DCM to extract (10mL×3), combine the organic phases, wash with 15mL saturated NaCl, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and pass the residue through a column Chromatographic 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°C, add 46mg EDCI, 33mg HOBT, 24mg TEA and 17mg compound 23-2. The reaction was stirred at RT for 5hrs under nitrogen protection. The reaction was completed by TLC detection, the reaction solution was poured into 5mL water, extracted with EtOAc (15mL×2), the organic phases were combined, washed with 15mL saturated NaCl three times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography 65 mg of compound 23-3 was obtained.

Step 4: Preparation of compound 23-4

65 mg of compound 23-3 was dissolved in 1.8 mL of THF, and MgBrCH ₃ (3M, 0.196 mL) was added dropwise at -78° C. The reaction solution was raised to 0° C. and stirred for 1 hrs. 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 purification yielded 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and 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 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 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(500MHz,DMSO-d ₆ )δ8.41(s,1H), 7.70(d,J=5.3Hz,1H), 7.30(d,J=6.8Hz,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 = 15.4Hz, 1H), 2.65 (d, J = 15.3Hz, 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 Pd ₂ (dba) ₃ , 6 mg Xantphos and 26 mg DIPEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 100°C and stirred for 5 hrs. 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 compound 24-6 was dissolved in 1.5 mL dioxane and 0.5 mL 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), and the solution was adjusted to pH=8 with saturated NaHCO _{3. The} solid precipitated out and 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(500MHz,DMSO-d ₆ )δ8.51(s,1H),8.35(s,1H),7.76(d,J=5.4Hz,1H),7.36(d,J=2.0Hz,2H) ,7.25-7.16(m,3H),6.16(d,J=5.3Hz,1H), 6.09(d,J=1.9Hz,1H), 5.49(t,J=5.9Hz,1H),4.54(d, J=5.7Hz,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=15.7Hz,1H ), 2.71(d,J=15.7Hz,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

500 mg of compound M20-SM and 186 mg of compound 25-1 were dissolved in 5 mL of DMSO, replaced with nitrogen three times, and the reaction was raised to 70°C under nitrogen protection and stirred for 4 hrs. TLC detected the completion of the reaction. The reaction solution was poured into 20mL water, extracted with EtOAc (30mL×2), combined the organic phases, washed with 30mL saturated NaCl three times, 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 was obtained.

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°C and stir for 12 hours. . 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

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

Step 4: Preparation of compound 25-5

32 mg of compound 25-4 and 50 mg of compound M22-2 were dissolved in 1 mL of dioxane, and 5 mg of Pd ₂ (dba) ₃ , 6 mg of Xantphos and 26 mg of DIPEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 100°C and 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 was precipitated, 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(500MHz,DMSO-d ₆ )δ8.34(s,1H),7.99(d,J=5.3Hz,1H), 7.31(d,J=6.9Hz,1H), 7.20-7.12(m, 3H), 6.40(d,J=5.4Hz,1H), 5.47(t,J=5.9Hz,1H), 4.53(d,J=5.8Hz,2H),3.93-3.88(m,3H),3.75- 3.72(m,4H),3.23-3.12(m,6H),3.06(d,J=15.6Hz,1H), 2.62(d,J=15.3Hz,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

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

Step 2: Preparation of compound 26-3

Under the protection of nitrogen, 0.30 g of compound 26-2 was dissolved in 10 mL of anhydrous dichloromethane, the temperature was reduced to -78° C., and DIBAL-H (1M, 2.6 mL) in n-hexane was added dropwise. The reaction was stirred at -78°C for 1 hour. Then slowly increase the temperature to -40°C and continue the reaction for 2 hours. TLC detected that the reaction was complete. At 0°C, 0.1 mL of water was slowly added dropwise, then sodium hydroxide aqueous solution (15%, 0.1 mL) was added dropwise, and then 0.25 mL of water was added. 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

80 mg of compound 26-3 and 52 mg of compound M-1 were dissolved in 4 mL of dioxane, 13 mg Pd ₂ (dba) ₃ , 16 mg Xantphos and 130 mg DIEA were added, and then reacted at 100° C. for 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 (2N, 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 sodium bicarbonate aqueous solution was added dropwise to adjust pH=9-10. The solid was filtered, 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(500MHz,DMSO-d ₆ )δ8.32(s,1H), 7.70–7.64(m,2H), 7.53(d,J=7.8Hz,1H), 7.42(d,J=7.8Hz, 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

400mg of compound M23-1 was dissolved in 20.0mL THF, cooled to 0°C in an ice bath, methylmagnesium bromide (2.0M, 2ml) was slowly added to the above solution, and the temperature was raised to room temperature and stirred for 0.5hrs. TLC detected that the reaction was complete. Saturated aqueous ammonium chloride solution (5 mL) was slowly added to the reaction solution to quench the reaction, EA (10 mL*3) was added for extraction, the organic phases were combined, washed with saturated brine (15 mL), and dried with anhydrous sodium sulfate. After filtration, 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, 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 was raised to 100°C and 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 the residue to dissolve it, 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(500MHz, DMSO-d ₆ )δ8.32(s,1H), 7.67(d,J=5.5Hz,1H), 7.31(d,J=7.0Hz,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=6.0Hz ,3H),1.17-1.13(m,1H).

The compounds 28-57 and 63 in Table 3 were prepared by using different reaction starting materials and appropriate reagents, using a method similar to that of the foregoing Example 26.

table 3

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 (500MHz, DMSO-d ₆ ): δ8.25 (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=5.1Hz,1H),6.12(t,J=4.5Hz,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=15.6Hz,1H), 2.82(d,J=4.5Hz,3H), 2.61(d,J=15.6Hz,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 (500MHz, DMSO-d ₆ ): δ8.11 (s, 1H), 7.31 (d, J = 6.5 Hz, 1H), 7.25-7.09 (m, 4H), 7.08-7.04 (m, 1H) ,6.89(d,J=7.8Hz,1H),5.39(t,J=5.6Hz,1H),4.50(d,J=5.5Hz,2H),3.85(s,3H),3.75-3.70(m, 2H),3.14-3.02(m,3H),2.61(d,J=15.6Hz,2H),1.91-1.81(m,1H),1.81-1.76(m,1H),1.53(d,J=12.9Hz ,1H),1.13(d,J=13.3Hz,1H). (Compound 33)

[M+H ⁺ ]=470.32

¹ H NMR (500MHz, DMSO-d ₆ ): δ8.66(d,J=4.7Hz,1H), 8.36(s,1H), 8.20(d,J=8.3Hz,1H), 8.05(d, J = 8.4 Hz, 1H), 7.84 (t, J = 7.6 Hz, 1H), 7.71 (t, J = 7.5 Hz, 1H), 7.32 (d, J = 6.6 Hz, 1H), 7.20-7.16 (m, 3H), 7.04 (d, J = 4.6 Hz, 1H), 5.49 (t, J = 5.8 Hz, 1H), 4.54 (d, J = 5.7 Hz, 2H), 4.01-3.80 (m, 3H), 3.21- 3.18(m,2H),3.07(d,J=15.7Hz,1H), 2.62(d,J=15.7Hz,1H),1.93-1.88(m,1H),1.83-1.78(m,1H),1.57 -1.54 (m, 1H), 1.17-1.13 (m, 1H). (Compound 35)

[M+H] ⁺ :488.3

¹ H NMR (500MHz, 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=5.6Hz,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 (500MHz, 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=15.6Hz,1H), 2.62(d,J=15.6Hz,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(500MHz,DMSO-d ₆ )δ8.31(s,1H), 7.67(d,J=5.6Hz,1H), 7.21-7.18(m,1H), 7.09(d,J=9.0Hz, 1H), 6.95(t,J=8.8Hz,1H), 6.36(s,2H),5.99-5.88(m,1H), 5.49(t,J=5.9Hz,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(500MHz,DMSO-d ₆ )δ8.31(s,1H), 7.67(d,J=5.4Hz,1H), 7.07(d,J=8.2Hz,1H), 6.90(m,1H) ,6.70(d,J=7.9Hz,1H),6.36(s,2H),5.91(d,J=5.3Hz,1H),5.48(m,1H),4.52(d,J=4.9Hz,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(500MHz,DMSO-d ₆ )δ8.32(s,1H),7.69-7.61(m,3H),7.41(t,J=7.6Hz,1H),6.35(s,2H),5.92( d,J=5.4Hz,1H),5.49(t,J=5.8Hz,1H),4.53(d,J=5.8Hz,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(500MHz,DMSO-d ₆ )δ8.31(s,1H), 7.67(d,J=6.0Hz,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=15.7Hz,1H), 2.60(d,J=15.7Hz,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(500MHz,DMSO-d ₆ )δ8.41-8.36(m,2H),8.32(s,1H),7.32(d,J=6.8Hz,1H),7.23-7.09(m,5H), 5.48(t,J=5.6Hz,1H),4.53(d,J=5.6Hz,2H),3.91-3.80(m,3H),3.21-3.13(m,2H),3.06(d,J=15.6Hz ,1H), 2.65(d,J=15.6Hz,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

13 mg of compound 21-2A was dissolved in 0.5 mL of MeOH, 3 mg of LiOH·H ₂ O in H ₂ O (0.1 mL) was added dropwise, and the reaction solution was heated to 40° C. and stirred for 12 hrs. TLC detects the completion of the reaction, 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 reduce pressure After concentration, the residue was purified by TLC plate to obtain 10 mg of compound 60-1.

Step 2: Preparation of compound 60-3

Dissolve 10 mg of compound 60-1 in 0.3 mL of THF and 0.3 mL of DCM, add 4 mg of EDCI, 3 mg of HOBT, and 2 mg of TEA, and stir for 1 hrs at RT. An additional 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

6mg of compound 60-3 was dissolved in 0.5mL MeOH, 2N HCl (0.1mL, methanol solution) was added, and the reaction was stirred at RT for 8hrs. 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

100mg of compound M23-1 and 57mg of compound 61-2 were dissolved in 2mL of tetrahydrofuran, 0.01ml of tetrabutylammonium fluoride was added, and then reacted for 2 hours at RT. TLC detected the complete reaction of the raw material, 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

61 mg of compound 61-3 and 50 mg of compound M1 were dissolved in 1.5 mL of dioxane, 10 mg Pd ₂ (dba) ₃ , 12 mg Xantphos, and 42 mg DIEA were added, and then reacted at 100° 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

Dissolve 35 mg of compound 61-4 in 1.5 mL of dioxane and 0.5 mL of methanol, add 0.14 mL of HCl in dioxane solution, and stir 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. 26 mg of compound 62-1 was added dropwise at 0°C and slowly warmed 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 over sodium sulfate and then desolventized, and purified by column chromatography to obtain 49 mg of compound 62-2.

Step 2: Preparation of compound 62-3

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 were dissolved in 2 mL of dioxane, replaced with nitrogen three times, and reacted at 100°C for 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

27 mg of compound 62-3 was dissolved in 1 mL of dioxane, and methanol solution of hydrochloric acid (2N, 0.4 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, 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 (500MHz, DMSO-d ₆ ,): δ8.35(s,1H), 7.67(d,J=5.4Hz,1H), 7.31(d,J=6.5Hz,1H), 7.18-7.15( m,3H),6.36(s,2H),5.88(d,J=5.1Hz,1H),4.45(s,2H),3.93-3.86(m,3H),3.25-3.15(m,2H),3.06 (d,J=15.7Hz,1H), 2.63(d,J=15.3Hz,1H),1.92-1.88(m,1H),1.82-1.78(m,1H),1.55(d,J=12.9Hz, 1H), 1.16 (d, J=13.8 Hz, 1H).

Example 64 Preparation of Compound 64:

Step 1: Preparation of compound 64-1

400 mg of compound M22-SM and 552 mg of compound M12 were dissolved in 8.0 mL of THF, 349 mg of N,N-diisopropylethylamine was added, and the reaction was stirred at 40°C for 16 hrs. 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 the protection of nitrogen, slowly drop 3.8 mL 1.0M DIBAL-H n-hexane solution at -78°C, and stir at -78°C for 15 min. LCMS detects that the reaction of the raw materials is complete. 0.5mL water and 0.5mL NaOH (2.5M) aqueous solution are added dropwise to quench at low temperature. After warming to room temperature, 0.5mL water is added dropwise. After filtration, the organic phase is 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 the protection of nitrogen, 56mg NaBH _{4 was} added at 0°C, and the reaction was stirred at room temperature for 2hrs. LCMS detected that the reaction was complete, 5mL saturated ammonium chloride was added dropwise under ice bath, returned to room temperature, concentrated in vacuo and spin-dried 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was heated to 70°C and 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 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. The residue was dissolved by adding H ₂ O (0.5 mL), adjusting the solution to pH=8-9 with saturated NaHCO ₃ , solids separated out, filtered, the filter cake solid was dissolved in DCM, dried by adding anhydrous sodium sulfate, filtered and concentrated in vacuo. 50.2 mg of compound 64 was obtained as a light yellow solid.

[M+H ⁺ ]=466.34.

¹ H NMR(500MHz,DMSO-d ₆ )δ8.33(s,1H), 7.96(d,J=5.0Hz,1H), 7.08(d,J=8.0Hz,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=15.0Hz,1H),2.54(d,J=15.0Hz,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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was heated to 70°C and 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 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(500MHz,DMSO-d ₆ )δ8.91(s,1H),8.51(d,J=5.5Hz,1H),8.36(s,1H),7.32(d,J=6.5Hz,1H) ,7.21-7.13(m,4H),5.30(t,J=6.0Hz,1H),4.53(s,2H),3.93-3.86(m,3H),3.33-3.16(m,2H),3.07(d ,J=15.0Hz,1H),2.64(d,J=15.5Hz,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. The reaction was stirred at 70°C 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 the protection of nitrogen, slowly add 4.0mL 1.0M DIBAL-H n-hexane solution dropwise at -78℃, and stir for 0.5hrs at -78℃. The reaction was completed by LCMS, 0.5mL water and 0.5mL 2.5M NaOH aqueous solution were added dropwise to quench at low temperature, and 0.5mL water was added dropwise after returning to room temperature. 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 the protection of nitrogen, 60mg NaBH _{4 was} added at 0°C, and the reaction was stirred at room temperature for 0.5 hrs. LCMS detected the completion of the reaction. The reaction was quenched by adding 5 mL of water dropwise under an ice bath. The temperature was restored to room temperature. After vacuum concentration and spin-drying of THF, 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction temperature is raised to 70°C and the reaction is 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(500MHz,DMSO-d ₆ )δ8.34(s,1H),7.96(d,J=5.0Hz,1H), 7.32(d,J=8.0Hz,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=16.0Hz,1H), 2.63(d,J=15.5Hz,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. The reaction was stirred at 70°C 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 the protection of nitrogen, slowly add 4.0mL 1.0M DIBAL-H n-hexane solution dropwise at -78℃, and stir for 0.5hrs at -78℃. The reaction was completed by LCMS, 0.5 mL of water and 0.5 mL of 2.5M NaOH aqueous solution were added dropwise to quench at low temperature, and 0.5 mL of water was added dropwise after returning to room temperature. After filtration, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 330 mg 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 the protection of nitrogen, 60mg NaBH _{4 was} added at 0°C, and the reaction was stirred at room temperature for 0.5 hrs. LCMS detected that the reaction was complete, and quenched by adding 5 mL of water dropwise under ice bath, returned 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 70°C and 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(500MHz,DMSO-d ₆ )δ8.34(s,1H),7.96(d,J=5.5Hz,1H),7.31(dd,J=5.5,8.0Hz,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=16.0Hz,1H), 2.63(d,J=16.0Hz,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. The temperature was raised to 100°C 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

Dissolve 0.7 g of compound 68-1 in 3 mL of THF, add 0.66 mg of tBuOK at room temperature, and stir for 0.5 hrs at room temperature. 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 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, and the reaction was stirred at RT for 1 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H ₂ O (4 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 (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(500MHz,CDCl3)δ8.36(s,1H),8.01(br,1H),7.34(d,J=6.5Hz,1H),7.26-7.21(m,3H),6.24(d,J =5.5Hz,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=16.0Hz,1H),2.89(br,3H),2.74(d,J=15.5Hz,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 N ₂ protection, dissolve 2.23g PPh ₃ in 8mL dioxane, add 1.15g NCS, stir at room temperature for 0.5hrs, the reaction solution will become thick white mud, add 0.7g compound M5, heat to 100℃, stir 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 the protection of nitrogen, 2.55mL 1.0M DIBAL-H n-hexane solution was slowly added dropwise at -78℃, and the reaction was stirred at -78℃ for 0.5hrs. LCMS detects that the reaction is complete. Add 0.1 mL of water and 0.1 mL of 2.5M NaOH aqueous solution to quench at low temperature. Return to room temperature and then add 0.25 mL of water. After filtration, the organic phase is 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 70°C and 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 nitrogen protection, 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 and react at room temperature for 1 hrs, TLC detects that the reaction is complete, the reaction solution is concentrated under reduced pressure . Add H ₂ O (4 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 (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 (500MHz, 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=5.5Hz,1H), 5.44(t,J=6.0Hz,1H), 4.48(d,J=5.5Hz,2H),3.96-3.83(m,3H),3.21-3.02(m, 2H),3.07(d,J=15.5Hz,1H),2.64(d,J=15.5Hz,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 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

Dissolve 3.5 g of compound 70-1, 3.82 g of compound M6 and 5.04 g of DIPEA in 40 mL of THF, and stir 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. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 100°C and 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 at RT and react for 1 hrs. TLC detects that the reaction is complete and the reaction solution is decompressed concentrate. 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 17.8 mg of light yellow solid compound 70.

[M+H+]=449.32

¹ H NMR (500MHz, 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 = 6.5Hz, 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).

Control 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-tyrosyl-phosphorylated peptide to its Src homology 2 (SH2) domain. The subsequent activation step results in the release of the SHP2 auto-inhibitory interface, which in turn activates the SHP2 protein tyrosine phosphatase (PTP) and can be used for substrate recognition and reaction catalysis. The surrogate DiFMUP was used to monitor the catalytic activity of SHP2 in the rapid fluorescence assay format.

experiment procedure:

(1) Compound preparation:

The compound of the present invention (10mM stock solution) was diluted to an appropriate multiple with 100% DMSO. 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 enzyme reaction working solution:

Perform SHP2 enzyme activity detection in a 96-well black polystyrene plate (flat bottom, low flange, non-binding surface) (Perki Elmer, Cat#6005270) at room temperature, using a final reaction volume of 50 μL and the following assay buffer conditions: 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 SHP2Activating 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 substitution substrate DiFMUP (Invitrogen, Cat#D6567) was added to the reaction, and the reaction was carried out 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%

Among them: Conversion_sample is the conversion rate reading of the sample; Conversion_min is the average value of the blank control well, representing the conversion rate reading of the wells without enzyme activity; Conversion_max is the average value of the positive control well ratio, representing the conversion rate reading of the wells without compound inhibition. The analysis software GraphPad Prism log (inhibitor) vs. response-Variable slope was used to fit the dose-response curve, and the IC ₅₀ value of the compound to the enzyme activity was calculated.

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

Table 4

化合物序号Compound number	化合物对SHP2酶活IC ₅₀(nM) Compound to SHP2 enzyme activity IC ₅₀ (nM)
化合物2Compound 2	8.568.56
化合物4Compound 4	3.193.19
化合物5Compound 5	12.6412.64
化合物7Compound 7	6.126.12
化合物8Compound 8	4.954.95
化合物9Compound 9	5.435.43
化合物10Compound 10	13.6413.64
化合物11Compound 11	13.8213.82
化合物12Compound 12	7.197.19
化合物13Compound 13	20.3720.37
化合物14Compound 14	20.6620.66
化合物15Compound 15	4.974.97
化合物16Compound 16	6.106.10
化合物17Compound 17	10.910.9
化合物18Compound 18	10.310.3
化合物19Compound 19	2.172.17
化合物20Compound 20	3.283.28
化合物21Compound 21	0.620.62
化合物22Compound 22	0.370.37
化合物23Compound 23	2.12.1
化合物24Compound 24	2.622.62
化合物25Compound 25	1.31.3
化合物26Compound 26	2.412.41
化合物27Compound 27	1.651.65
化合物29Compound 29	2.02.0
化合物30Compound 30	0.780.78
化合物31Compound 31	1.01.0

化合物32Compound 32	1.471.47
化合物33Compound 33	0.90.9
化合物34Compound 34	1.71.7
化合物35Compound 35	0.70.7
化合物36Compound 36	1.191.19
化合物37Compound 37	1.21.2
化合物38Compound 38	0.60.6
化合物39Compound 39	0.710.71
化合物40Compound 40	1.801.80
化合物41Compound 41	0.810.81
化合物42Compound 42	0.850.85
化合物43Compound 43	2.412.41
化合物44Compound 44	0.830.83
化合物45Compound 45	1.601.60
化合物46Compound 46	1.101.10
化合物47Compound 47	0.600.60
化合物48Compound 48	1.71.7
化合物49Compound 49	4.14.1
化合物50Compound 50	0.70.7
化合物51Compound 51	//
化合物52Compound 52	//
化合物53Compound 53	22
化合物54Compound 54	11
化合物55Compound 55	9.39.3
化合物56Compound 56	6.86.8
化合物57Compound 57	//
化合物58Compound 58	0.720.72
化合物59Compound 59	2.152.15
化合物60Compound 60	7.197.19
化合物61Compound 61	19.7519.75
化合物62Compound 62	1.41.4
化合物63Compound 63	4.34.3
化合物64Compound 64	2.02.0
化合物65Compound 65	2.12.1
化合物66Compound 66	3.13.1
化合物67Compound 67	2.02.0
化合物68Compound 68	1.81.8
化合物69Compound 69	4.74.7
化合物70Compound 70	2.32.3

Note: "/" means not tested.

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

Example B: Cell Proliferation Test

An in vitro cell assay was 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, it is an indicator of living cell metabolism. Its content directly reflects the number and cell state of cells. During the experiment, CellTiter-Glo ^TM reagent was added to the cell culture medium to measure the luminescence value. The value is directly proportional to the amount of ATP, and ATP is positively related to the number of living cells, so cell viability can be inspected by detecting ATP content.

experiment procedure:

(1) Cell plating:

Take a bottle of KYSE-520 cells in logarithmic growth phase, digest the resuspended cells, count them, adjust the cell density and inoculate them into a 96-well plate, inoculate 1000 cells per well, and place the well plate at 37℃, 5% CO ₂ 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 logarithmic growth phase, digest and resuspend the cells, count them, adjust the cell density and inoculate them in a 96-well plate, inoculate 4000 cells per well, and place the plate at 37°C, 5% CO _2. After culturing in the incubator for 24hrs, adding the compound of the present invention for treatment;

(2) Cell compound treatment:

Prepare an 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. Enter 37°C, 5% CO ₂ incubator for 120hrs. Only adding medium without adding cell wells is set as the zero adjustment group; the compound concentration of 0 nM group is the blank group.

(3) CTG detection:

After cell culture for 120hrs, add 50μL to each well

Luminescent Cell Viability Assay solution, gently shake for 2mins, continue to incubate at room temperature for 10mins, 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

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 the hERG potassium channel was cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg/mL G418 at a culture temperature of 37°C and a carbon dioxide concentration of 5%.

Cell passage: Remove the old medium and wash once with PBS, then add 1mL TrypLE ^TM 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. The cell suspension was transferred to a sterile centrifuge tube and centrifuged at 1000 rpm for 5 minutes to collect the cells. To expand or maintain the culture, inoculate the cells in 6 cm cell culture dishes, each cell culture dish, the amount of cells inoculated is 2.5*105 cells (final volume: 5mL).

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

For patch clamp detection, the cells were separated with TrypLE ^TM Express before the experiment, and 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 fluid

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

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

Compound formulation

Dilute the test compound stock solution with extracellular fluid to prepare a 10 μM working solution, or a gradient dilution to a solution of 0.3 μM, 1 μM, 3 μM, 10 μM, 30 μM.

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

Cisapride (positive control)

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

The cisapride stock solution was diluted successively with DMSO at a 10-fold dilution from high to low to a 10μM 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 -80mV. The clamping voltage is depolarized from -80mV to -50mV for 0.5 seconds, then stepped to 30mV for 2.5 seconds, and then quickly restored to -50mV for 4 seconds to stimulate the tail current of the hERG channel. Collect data repeatedly every 10 seconds to observe the effect of the drug on the hERG tail current. A stimulus of -50mV for 0.5 seconds was used as leakage current detection. The experimental data is 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 contact the recording electrode on 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 cover glass covered with cells 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 out 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 ≤20MΩ

(2) Sealing resistance≥1GΩ

(3) Peak tail current peak ≥400pA

(4) The initial tail current peak is greater than the activation current peak

(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 -80mV (leakage current ≤100pA)

data analysis

First, the current after each drug concentration is normalized to the blank control current

Then calculate the inhibition rate corresponding to each drug concentration

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

Table 6

编号Numbering	抑制率@10μMInhibition rate @10μM
对照例1Comparative example 1	95.12％95.12%
对照例2Comparative example 2	>95.6％>95.6%
化合物3Compound 3	86.7％86.7%
化合物5Compound 5	60.8％60.8%
化合物8Compound 8	97.36％97.36%
化合物9Compound 9	93.35％93.35%
化合物16Compound 16	87.99％87.99%
化合物18Compound 18	42.63％42.63%
化合物21Compound 21	82.3％82.3%
化合物22Compound 22	43.5％43.5%
化合物24Compound 24	79.9％79.9%
化合物26Compound 26	>90％>90%
化合物30Compound 30	92.2％92.2%
化合物33Compound 33	>90％>90%
化合物39Compound 39	83.7％83.7%
化合物40Compound 40	98.1％98.1%
化合物41Compound 41	85.6％85.6%

化合物42Compound 42	87.6％87.6%
化合物43Compound 43	56.6％56.6%
化合物44Compound 44	80.2％80.2%
化合物45Compound 45	78.0％78.0%
化合物46Compound 46	87.8％87.8%
化合物58Compound 58	69.4％69.4%
化合物59Compound 59	57.0％57.0%
化合物62Compound 62	>90％>90%
化合物64Compound 64	5.02％5.02%
化合物65Compound 65	62.1％62.1%
化合物66Compound 66	37.1％37.1%
化合物67Compound 67	29.5％29.5%
化合物68Compound 68	54.4％54.4%
化合物69Compound 69	26.3％26.3%
化合物70Compound 70	9.12％9.12%

We unexpectedly found that while the compounds of the present invention generally have 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 formula I is a hydroxyl group. A methyl group and both A ₁ and A ₃ positions are N, such as compound 22, compounds 64-69; 2), in formula I, the R ₁ position on the pyrazine ring is an amide group and 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 100mM sodium phosphate and 150mM 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 12.0g/L NaH ₂ PO ₄ and 8.77g/L NaCl in an acidic solution, which can be stored at 4°C for 7 days. The alkaline solution was titrated to pH 7.4 with an acidic solution and stored at 4°C for 7 days. Check on the day of the experiment and adjust if the pH exceeds the specification of 7.4±0.1.

2) Preparation of plasma

The frozen plasma was immediately thawed at room temperature.

The plasma was centrifuged at 3,220g 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). Only use plasma within the range of pH 7 to pH 8.

3) Preparation of working solution

Working solutions of the test compound and the control compound ketoconazole were prepared with DMSO at a concentration of 400 μM. Then 4 μL of the working solution was removed and mixed with 796 μL of human, dog, monkey, rat, or mouse plasma to finally 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 add 400 μL of dialysis buffer (PBS) on the opposite side for dialysis. The determination is performed 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, the seal was removed, and 50 μL of samples from the buffer chamber and the plasma chamber were transferred 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 of 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 samples in the plate were vortexed for 5 minutes and centrifuged at 3220 g for 30 minutes at 4°C. Then transfer 100 μL of supernatant with 100 μL or 200 μL of water to a new 96-well plate for LC-MS/MS analysis (depending on 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 free part

Peak area ratio _{plasma chamber} indicates the concentration of free and bound fraction

The peak area ratio to the _{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

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 with 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 appended claims of the present invention.

Claims

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

among them,

Optionally R 1 is 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;

Optionally R 2 is selected from hydrogen, C 1-4 alkyl group or a substituted C 1-4 alkyl;

Optionally 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 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;

Optionally R 4 is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, hydroxymethyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl base;

A 1 is arbitrarily selected from CR 5 or N;

A 2 is arbitrarily selected from CR 6 or 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, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted 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, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents; 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 a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group 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 containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;

n is 0, 1, 2, 3, or 4.
The compound according to claim 1, wherein, R & lt an arbitrarily selected from amino, -C (O) NH 2, -C≡N, C 1-3 alkyl group, a substituted group containing C 1-3 alkoxy Group or C 1-3 alkoxy.
The compound according to claim 1 or 2, wherein R 1 is arbitrarily selected from amino group, -C(O)NH 2 , -C≡N, hydroxy-substituted methyl,
The compound of any one of claims 1-3, wherein R 2 is hydrogen.
A compound according to any one of claims 1-4, wherein, R 3 is optionally selected from hydrogen, halo, C 1-3 alkyl or an optionally substituted C 1-3 alkyl.
The compound according to any one of claims 1-5, wherein R 3 is arbitrarily selected from hydrogen, chlorine or methyl.
The compound according to any one of claims 1-6, wherein R 4 is 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 1-7, wherein R 4 is arbitrarily selected from hydrogen, F, Cl, -CHF 2 , CF 3 or -O-CH 3 .
The compound according to any one of claims 1-8, wherein A 1 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 1-9, 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 1-10, wherein A 2 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 1-11, wherein A 2 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 1-12, wherein A 3 is CR 7 or N, wherein R 7 is hydrogen or halogen.
The compound according to any one of claims 1-13, 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 heterocyclyl.
The compound according to any one of claims 1-14, 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 of any one of claims 1-15, wherein U is CH 2 or O.
The compound according to any one of claims 1-16, wherein ring A is arbitrarily selected from phenyl or C 5-6 heteroaryl, and the C 5-6 heteroaryl contains one or two N Or S heteroatom.
The compound according to any one of claims 1-17, wherein Rx is 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 1-18, wherein Rx is arbitrarily selected from hydrogen, OH, F, Cl, Br, -CN, trifluoromethyl or methoxy.
The compound according to any one of claims 1-19, 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 of the formula II shows:

among them,

Optionally 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 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;

A 1 is arbitrarily selected from CR 5 or N;

U is arbitrarily 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, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl Group or C 5-6 heterocycle; or,

L is selected from S;

R x 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 group, a substituted group containing C 1-8 alkoxy, C 3-8 cycloalkyl or C 3-8 cycloalkyl group having substituent;

n is 0, 1, 2, 3, or 4.
The compound of claim 21, wherein R 3 is arbitrarily selected from hydrogen or methyl.
The compound of claim 21 or 22, wherein A 1 is CR 5 , wherein R 5 is selected from F or Cl.
The compound of claim 21 or 22, wherein A 1 is N.
The compound according to any one of claims 21-24, wherein U is O or CH 2 .
The compound according to any one of claims 21-25, wherein R x is optionally 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 21-26, wherein R x is arbitrarily selected from hydrogen, OH, CN, Cl, F, trifluoromethyl or methoxy.
The compound of any one of claims 21-27, 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 of the formula As shown in III:

Optionally 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 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;

Optionally R 4 is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, A substituted C 1-8 alkoxy group, a C 2-8 alkenyl group, a substituted C 2-8 alkenyl group, a C 2-8 alkynyl group or a substituted C 2-8 alkynyl group;

A 1 is arbitrarily selected from CR 5 or N;

A 2 is arbitrarily selected from CR 6 or N;

A 3 is arbitrarily selected from CR 7 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, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted 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, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents;

L is selected from S;

Ring A is optionally selected from a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group 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 containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;

n is 0, 1, 2, 3, or 4.
The compound of claim 29, wherein R 3 is selected from hydrogen.
The compound according to claim 29 or 30, wherein R 4 is arbitrarily selected from hydrogen or Cl.
The compound according to any one of claims 29-31, 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 29-32, wherein A 2 is CR 6 or N, wherein R 6 is selected from Cl, amino,
Or -O-CH 3 .
The compound of any one of claims 29-33, wherein A 3 is CR 7 or N, wherein R 7 is hydrogen.
The compound of any one of claims 29-34, wherein U is CH 2 .
The compound according to any one of claims 29-35, wherein ring A is optionally selected from phenyl or C 5-6 heteroaryl, said C 5-6 heteroaryl contains one or two N Or S heteroatom.
The compound according to any one of claims 29-36, wherein Rx is arbitrarily selected from hydrogen, Cl, Br, trifluoromethyl or methoxy.
The compound according to any one of claims 29-37, wherein:
Selected from
The compound of claim 1, wherein the compound is:

2) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)-5-methylpyrazin-2-yl)methanol;

3) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine) -4-yl)thio)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-chloropyridine) -4-yl)thio)pyrazine-2-carboxamide;

7) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methoxy (Pyridin-4-yl)thio)pyrazine-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-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'-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'-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-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2, 4'-Piperidine]-1'-yl)pyrazine-2-carboxamide;

16)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((6-amino-3-chloropyridine -2-yl)thio)pyrazine-2-carboxamide;

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-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-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-fluoro-3-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;

20)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-methoxypyrimidine- 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-chloro Pyridin-4-yl)thio)pyrazin-2-yl)methanol;

22)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-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 -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-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-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-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-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-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'-piperidine]-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-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;

34)(S)-(3-(1-amino-1,3-dihydrospiro[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-ylsulfanyl )Pyrazin-2-yl)methanol;

36)(S)-(3-(1-amino-1,3-dihydrospiro[indene-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'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)pyrazin-2-yl)methanol;

38)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-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-fluoro Pyridin-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-methyl Oxypyridin-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)pyrazin-2-yl)methanol;

43)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-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'-Piperidine]-1'-yl)pyrazin-2-yl)methanol;

47)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(pyridin-4-ylsulfanyl) Pyrazin-2-yl)methanol;

48)(S)-(6-((2-amino-3-chloropyridin-4-yl)thio)-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-dihydro Spiro[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)pyridine (Azin-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'-piperidine]-1'-yl)-6-((2,3-dihydro- [1,4]Dioxo[2,3-b]pyridin-8-yl)thio)pyrazin-2-yl)methanol;

54)(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-nitrile;

55)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-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-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl;

57)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-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-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'-piperidine]-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-ol;

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'-piperidine]-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) Pyrazin-2-yl)methanol;

66)(S)-(3-(1-Amino-5-chloro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;

67)(S)-(3-(1-amino-5-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;

68)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(methylamino) Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;

69)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-4- Yl)thio)-5-methylpiperazin-2-yl)methanol; or

70)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((2-aminopyrimidin-4-yl ) Thio) pyrazine-2-carboxamide.
A pharmaceutical composition characterized by comprising a therapeutically effective amount of at least one compound according to any one of claims 1-38 and at least one pharmaceutically acceptable excipient.
The pharmaceutical composition according to claim 40, wherein the mass percentage of the compound and pharmaceutically acceptable excipients is 0.0001:1-10.
Use of the compound of any one of claims 1-39 or the pharmaceutical composition of claim 40 or 41 in the preparation of medicines.
The application according to claim 42, wherein the medicine is used to treat, prevent, delay or stop cancer, cancer metastasis, cardiovascular disease, immune disease, fibrosis or eye disease.
Use of the compound of any one of claims 1-39 or the pharmaceutical composition of claim 40 or 41 in the preparation of a medicine for treating diseases mediated by SHP2.
The use according to claim 44, wherein the disease is cancer.
The use according to claim 43 or 45, wherein the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell Cancer, acute myeloid leukemia, breast cancer, esophageal tumor, lung cancer, colon cancer, head cancer, gastric cancer, lymphoma, glioblastoma, pancreatic cancer, or a combination thereof.
The use according to claim 42, wherein the drug is used as an SHP2 inhibitor.
A method for treating and/or preventing diseases mediated by SHP2, characterized in that a therapeutically effective amount of the compound according to any one of claims 1-39 or the drug according to claim 40 or 41 is administered to a subject to be treated combination.
The method of claim 48, wherein the SHP2-mediated disease is cancer.
The method of claim 49, 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, gastric cancer, lymphoma, glioblastoma, pancreatic cancer, or a combination thereof.
A method for treating cancer, comprising administering a therapeutically effective amount of the compound of any one of claims 1-38 or the pharmaceutical composition of claim 40 or 41 to a subject, wherein the cancer is 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, Gastric cancer, lymphoma, glioblastoma, pancreatic cancer, or a combination thereof.
The method according to any one of claims 48-51, wherein the subject to be treated is a human.