CN114057770A - Bifunctional compounds targeting EGFR protein degradation - Google Patents

Abstract

The present invention provides a novel bifunctional compound, or a stereoisomer, a pharmaceutically acceptable salt thereof, and its use in preparing a medicine. The bifunctional compound of the present invention is a protein targeting combination, which can selectively promote the ubiquitination and degradation of EGFR protein, and can be used for the treatment of various cancers/tumors.

Description

Bifunctional compounds targeting EGFR protein degradation

technical field

The invention belongs to the field of medicine, in particular to a bifunctional compound targeting EGFR protein degradation and its use in preparing medicine.

Background technique

EGFR (Epidermal Growth Factor Receptor) is a member of the transmembrane protein tyrosine kinase erbB receptor family. EGFR can form homodimers on the cell membrane by binding to its ligands, such as epidermal growth factor (EGF), or heterodimers with other receptors in the family (such as erbB2, erbB3, or erbB4) . The formation of these dimers can cause phosphorylation of key tyrosine residues in EGFR cells, thereby activating multiple downstream signaling pathways in cells. These intracellular signaling pathways play important roles in cell proliferation, survival and anti-apoptosis. Dysregulation of the EGFR signaling pathway, including increased expression of ligands and receptors, EGFR gene amplification, and mutation, can promote the malignant transformation of cells and play an important role in tumor cell proliferation, invasion, metastasis, and angiogenesis. Therefore, EGFR is considered as a target for anticancer drug development.

Ubiquitin itself consists of 76 amino acid residues with a molecular weight of about 8.5kDa. It is named "ubiquitin" because it is widely present in eukaryotic organisms. Ubiquitination refers to the process in which ubiquitin molecules classify intracellular proteins under the action of a series of special enzymes, select target protein molecules, and specifically modify the target protein to form target protein polyubiquitination. prime chain. These special enzymes include ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), ubiquitin ligase (E3), etc. Ubiquitination is closely related to the pathogenesis of tumors, cardiovascular, autoimmunity and other diseases. Degradation of proteins by the ubiquitin-proteasome system is an important pathway for selective protein degradation, which can be simply divided into three steps: first, a ligase adds a ubiquitin tag to the target protein, and then after multiple rounds of ubiquitination With multiple ubiquitin tags, the polyubiquitinated protein is recognized by a proteasome and degraded. By constructing a bifunctional compound, one end is a structure targeting the target protein; the other end is a protein degradation system such as E3 ligase, and the middle is connected by a suitable connecting chain, which can effectively promote the degradation of the target protein.

Most of the marketed EGFR inhibitor drugs develop resistance to small molecule inhibitors after treatment. The drug resistance mechanisms include EGFR secondary mutations, bypass pathway activation and so on. There are no drugs on the market that degrade EGFR proteins by constructing bifunctional compounds. The development of bifunctional compounds that can degrade EGFR protein by ubiquitination may become a new method for the treatment of tumors or cancers, which has broad application prospects.

SUMMARY OF THE INVENTION

The present invention provides a bifunctional compound shown in formula I, or a stereoisomer or a pharmaceutically acceptable salt thereof:

in,

R ¹ is selected from hydrogen, halogen, =O, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3-10 yuan cycloalkyl), -C _0-2 alkylene-(3-10 membered heterocycloalkyl);

R ² is selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3 ~10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

X ¹ and X ² are independently selected from O, S, NR ^x1 , CR ^x1 R ^x2 ;

R ^x1 and R ^x2 are independently selected from hydrogen, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), - C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Y is selected from N or CR ^Y1 ;

R ^Y1 is selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _0-2 alkylene base-(3-10 membered heterocycloalkyl);

Ring A is selected from benzene ring and 5-6 membered aromatic heterocycle; wherein benzene ring and aromatic heterocycle can be further substituted by one, two or three R ^A ;

Each R ^A is independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene base-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

R ⁴ and R ⁵ are independently 0, 1, 2 or 3 respectively;

R ⁴ and R ⁵ are independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene Alkyl-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Z ¹ , Z ² , Z ³ , Z ⁴ are independently selected from CH or N;

Ring B is selected from 5-6 membered cycloalkyl, 5-6 membered heterocycloalkyl, 5-6 membered aromatic ring, 5-6 membered aromatic heterocycle; wherein cycloalkyl, heterocycloalkyl, aromatic ring, aromatic ring The heterocycle may be further substituted with one, two or three ^R ;

Each R ^B is independently selected from hydrogen, halogen, cyano, nitro, =O, =S, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, - C _0-2 alkylene-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

L is -(L ^A ) _q -;

q is an integer greater than or equal to 1, and an integer less than or equal to 100;

Each LA is independently selected from CRR, ^C (O), O, S, S(O), S(O) ₂ , NR, -CR=CR-, -C≡C-, P(O)R, P(O)OR, 3-10-membered cycloalkane, 3-10-membered heterocycloalkane, 5-10-membered aromatic ring, 5-10-membered aromatic heterocycle, 5-12-membered spirocycle, 5-12-membered spiroheterocycle , 5-12-membered bridged ring, 5-12-membered bridged heterocyclic ring; wherein cycloalkane, heterocycloalkane, aromatic ring, aromatic heterocyclic ring, spirocyclic ring, spiro heterocyclic ring, bridged ring, bridged heterocyclic ring can be further replaced by one, two one or three R ^LA substitutions;

Each R ^LA is independently selected from hydrogen, halogen, =O, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _{0- 2} alkylene-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Each R is independently selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _{0- 2} -alkylene-(3- to 10-membered heterocycloalkyl).

further,

或

Ring B is selected from

or

further,

L is -L ^A1 -L ^A2 -L ^A3 -L ^A4 -L ^A5 -L ^A6 -L ^A7 -;

相连，另一端与L^A2相连；Among them, one end of L ^A1 is connected to

connected, and the other end is connected to L ^A2 ;

相连；One end of L ^A7 is connected to L ^A6 , and the other end is connected to

connected;

L ^A1 is selected from -(C _0-2 alkylene)-;

L ^A2 is selected from -NR-, -(5-6 membered heterocycloalkyl)- or none;

L ^A3 is selected from -(5-6-membered heterocycloalkyl)-, -(5-10-membered bridged heterocycle)-, -(6-9 membered spiro heterocycle)-, -(C _1-10 alkylene )-NR-, -(5~6 membered heterocycloalkyl)-NR-, -(5~10 membered bridged heterocycle)-NR-, -(6~9 membered spiro heterocycle)-NR-, -( C _0～10 alkylene)-;

L ^A4 is selected from -C(O)-, -(5-6 membered heterocycloalkyl)- or none;

L ^A5 is selected from -(C _0-10 alkylene)- or -C(O)-;

L ^A6 is selected from -O-, -NR- or none;

L ^A7 is selected from -(CH≡CH)-, -(3-6 membered cycloalkane)-, -(3-6 membered heterocycle)-, -(5-6 membered aromatic heterocycle)-, -(6- 9-membered spirocycle)-, -(6-9 membered spiroheterocycle)-, -(C _0-10 alkylene)- or none;

Each R is independently selected from hydrogen, C _1-10 alkyl.

go a step further,

L ^{A3 is} selected from

或无。L ^A7 is selected from -(CH≡CH)-, -(C _0-10 alkylene)-,

or none.

further,

R ¹ is selected from methyl; R ² is selected from methyl; X ¹ is selected from O; X ² is selected from CH ₂ ; Y is selected from N;

Ring A is selected from pyrazole or pyridine; wherein pyrazole or pyridine can be further substituted by one, two or three ^RAs ; each ^RA is independently selected from hydrogen, methyl;

R ⁴ and R ⁵ are each independently selected from hydrogen.

Further, the compound of the formula I is shown in the formula IIa:

in,

R ¹ is selected from hydrogen, halogen, =O, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3-10 yuan cycloalkyl), -C _0-2 alkylene-(3-10 membered heterocycloalkyl);

R ² is selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3 ~10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

X ¹ and X ² are independently selected from O, S, NR ^x1 , CR ^x1 R ^x2 ;

R ^x1 and R ^x2 are independently selected from hydrogen, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), - C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Y is selected from N or CR ^Y1 ;

R ^Y1 is selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _0-2 alkylene base-(3-10 membered heterocycloalkyl);

Ring A is selected from benzene ring and 5-6 membered aromatic heterocycle; wherein benzene ring and aromatic heterocycle can be further substituted by one, two or three R ^A ;

Each R ^A is independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene base-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

R ⁴ and R ⁵ are independently 0, 1, 2 or 3 respectively;

R ⁴ and R ⁵ are independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene Alkyl-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Z ¹ , Z ² , Z ³ , Z ⁴ are independently selected from CH or N;

L is -(L ^A ) _q -;

q is an integer greater than or equal to 1;

Each LA is independently selected from CRR, ^C (O), O, S, S(O), S(O) ₂ , NR, -CR=CR-, -C≡C-, P(O)R, P(O)OR, 3-10-membered cycloalkyl, 3-10-membered heterocycloalkyl, 5-10-membered aromatic ring, 5-10-membered aromatic heterocycle, 5-12-membered spiro, 5-12-membered spiro Heterocycle, 5-12-membered bridged ring, 5-12-membered bridged heterocycle; wherein cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle, spirocycle, spiroheterocycle, bridged ring, bridged heterocycle can be further replaced by one, two or three R ^LAs ;

Each R ^LA is independently selected from hydrogen, halogen, =O, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _{0- 2} alkylene-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Each R is independently selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _{0- 2} -alkylene-(3- to 10-membered heterocycloalkyl).

Further, the compound of formula I is shown in formula IIb:

in,

R ¹ is selected from hydrogen, halogen, =O, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3-10 yuan cycloalkyl), -C _0-2 alkylene-(3-10 membered heterocycloalkyl);

R ² is selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3 ~10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

X ¹ and X ² are independently selected from O, S, NR ^x1 , CR ^x1 R ^x2 ;

R ^x1 and R ^x2 are independently selected from hydrogen, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), - C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Y is selected from N or CR ^Y1 ;

R ^Y1 is selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _0-2 alkylene base-(3-10 membered heterocycloalkyl);

Ring A is selected from benzene ring and 5-6 membered aromatic heterocycle; wherein benzene ring and aromatic heterocycle can be further substituted by one, two or three R ^A ;

Each R ^A is independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene base-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

R ⁴ and R ⁵ are independently 0, 1, 2 or 3 respectively;

R ⁴ and R ⁵ are independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene Alkyl-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Z ¹ , Z ² , Z ³ , Z ⁴ are independently selected from CH or N;

R ^B is selected from hydrogen, halogen, cyano, nitro, =O, =S, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 Alkylene-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

L is -(L ^A ) _q -;

q is an integer greater than or equal to 1;

Each LA is independently selected from CRR, ^C (O), O, S, S(O), S(O) ₂ , NR, -CR=CR-, -C≡C-, P(O)R, P(O)OR, 3-10-membered cycloalkyl, 3-10-membered heterocycloalkyl, 5-10-membered aromatic ring, 5-10-membered aromatic heterocycle, 5-12-membered spiro, 5-12-membered spiro Heterocycle, 5-12-membered bridged ring, 5-12-membered bridged heterocycle; wherein cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle, spirocycle, spiroheterocycle, bridged ring, bridged heterocycle can be further replaced by one, two or three R ^LAs ;

Each R ^LA is independently selected from hydrogen, halogen, =O, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _{0- 2} alkylene-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Each R is independently selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _{0- 2} -alkylene-(3- to 10-membered heterocycloalkyl).

Further, the compound of formula I is shown in formula IIc:

in,

R ¹ is selected from hydrogen, halogen, =O, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3-10 yuan cycloalkyl), -C _0-2 alkylene-(3-10 membered heterocycloalkyl);

R ² is selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene-(3 ~10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

X ¹ and X ² are independently selected from O, S, NR ^x1 , CR ^x1 R ^x2 ;

R ^x1 and R ^x2 are independently selected from hydrogen, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), - C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Y is selected from N or CR ^Y1 ;

R ^Y1 is selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _0-2 alkylene base-(3-10 membered heterocycloalkyl);

Ring A is selected from benzene ring and 5-6 membered aromatic heterocycle; wherein benzene ring and aromatic heterocycle can be further substituted by one, two or three R ^A ;

Each R ^A is independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene base-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

R ⁴ and R ⁵ are independently 0, 1, 2 or 3 respectively;

R ⁴ and R ⁵ are independently selected from hydrogen, halogen, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _0-2 alkylene Alkyl-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Z ¹ , Z ² , Z ³ , Z ⁴ are independently selected from CH or N;

W is selected from C(O), CRR, NR;

L is -(L ^A ) _q -;

q is an integer greater than or equal to 1;

Each LA is independently selected from CRR, ^C (O), O, S, S(O), S(O) ₂ , NR, -CR=CR-, -C≡C-, P(O)R, P(O)OR, 3-10-membered cycloalkyl, 3-10-membered heterocycloalkyl, 5-10-membered aromatic ring, 5-10-membered aromatic heterocycle, 5-12-membered spiro, 5-12-membered spiro Heterocycle, 5-12-membered bridged ring, 5-12-membered bridged heterocycle; wherein cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle, spirocycle, spiroheterocycle, bridged ring, bridged heterocycle can be further replaced by one, two or three R ^LAs ;

Each R ^LA is independently selected from hydrogen, halogen, =O, cyano, nitro, -C _1-6 alkyl, halogen-substituted -C _1-6 alkyl, -OR, -NRR, -C _{0- 2} alkylene-(3-10-membered cycloalkyl), -C _0-2 alkylene-(3-10-membered heterocycloalkyl);

Each R is independently selected from hydrogen, C _1-10 alkyl, halogen-substituted -C _1-6 alkyl, -C _0-2 alkylene-(3-10 membered cycloalkyl), -C _{0- 2} -alkylene-(3- to 10-membered heterocycloalkyl).

More specifically,

L is selected from the following groups:

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-(CH≡CH)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-(C _0-10 alkylene)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-(3-6 membered cycloalkane)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-(3-6 membered heterocycle)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-(5-6 membered aromatic heterocycle)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-(5-6 membered aromatic hetero ring)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene base)-(5-6 membered aromatic heterocycle)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-O-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-O-(3-6 membered cycloalkane)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-O-(C _0-10 alkylene)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-10 alkylene)-(6-9 membered spiroheterocycle)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-C(O)-(C _0-10 alkylene)-(CH≡CH)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-C(O)-(C _0-10 alkylene)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(5-6 membered heterocycloalkyl)-C(O)-(C _0-10 alkylene)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-C(O)-(C _0～2 alkylene)-O-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-C(O)-(C _0-10 alkylene)-(3-6 membered cycloalkane)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-C(O)-(C _0-10 alkylene)-(3-6 membered heterocycle)-,

-(C _0-2 alkylene)-(5-6 membered heterocycloalkyl)-C(O)-(C _0-10 alkylene)-(5-6 membered aromatic heterocycle)-.

More specifically,

R ¹ is selected from methyl; R ² is selected from methyl; X ¹ is selected from O; X ² is selected from CH ₂ ; Y is selected from N;

Ring A is selected from pyrazole or pyridine; wherein pyrazole or pyridine can be further substituted by one, two or three ^RAs ; each ^RA is independently selected from hydrogen, methyl;

R ⁴ and R ⁵ are independently selected from hydrogen;

W is selected from C(O), _CH2 , NH;

R ^B is selected from methyl;

L is selected from the following linking groups:

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-(CH≡CH)-,

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-,

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-(C _0-10 alkylene)-,

-(C _0-1 alkylene)-(piperazine)-(piperazine)-(C _0-10 alkylene)-,

-(C _0-1 alkylene)-(piperazine)-(piperazine)-(C _0-10 alkylene)-(pyrazole)-,

-(C _0-1 alkylene)-(piperazine)-(C _0-2 alkylene)-(piperazine)-(C _0-10 alkylene)-(pyrazole)-,

-(C _0-1 alkylene)-piperazine)-(cyclobutane)-,

-(C _0-1 alkylene)-(piperazine)-(cyclobutylamine)-,

-(C _0-1 alkylene)-(1.2.5.6-tetrahydropyridine)-(C _0-10 alkylene)-,

-(C _0-1 alkylene)-(piperidine)-(C _0-10 alkylene)-,

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-(piperidine)-

-(C _0-1 alkylene)-(piperazine)-(piperazine)-C(O)-(C _0-10 alkylene)-

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-(tetrahydropyrrole)-C(O)-(C _0-10 alkylene)-O-,

-(C _0-1 alkylene)-(piperazine)-C(O)-(C _0-10 alkylene)-(CH≡CH)-,

-(C _0-1 alkylene)-(piperazine)-C(O)-(C _0-10 alkylene)-,

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-(pyrazole)-,

-(C _0-1 alkylene)-(piperazine)-C(O)-(C _0-10 alkylene)-(pyrazole)-,

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-O-,

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-O-(C _1-10 alkylene),

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-O-(3-4 membered cycloalkane)-,

-(C _0-1 alkylene)-(piperazine)-(C _0-10 alkylene)-(2-azaspiro[3.3]heptane)-.

More specifically, L is selected from the following linking groups:

相连；上述结构中的2端与

相连。In the above structure, the 1 end and

Connected; 2 ends in the above structure are connected with

connected.

In some specific embodiments of the present invention, the compound is specifically:

The present invention also provides the use of the above-mentioned bifunctional compound, or a stereoisomer or a pharmaceutically acceptable salt thereof, in the preparation of a medicament for the treatment of tumor or cancer.

The present invention also provides a composition comprising any of the above-mentioned bifunctional compounds, or a stereoisomer thereof, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

The compounds and derivatives provided in the present invention may be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

Definitions of terms used in the present invention: Unless otherwise specified, the initial definitions of groups or terms provided herein apply to the group or term throughout the specification; for terms that are not specifically defined herein, the content and context of the disclosure should be used. , give their meanings that those skilled in the art can give them.

"Substitution" means that a hydrogen atom in a molecule is replaced by a different atom or molecule.

"May be further substituted" means that "substitution" may but need not occur, and the description includes instances where it occurs or not.

Minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, eg, the prefix C _a-b alkyl denotes any alkyl group containing "a" to "b" carbon atoms. Thus, for example, "C _1-4 alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

"Alkyl" refers to a saturated hydrocarbon chain having the specified number of member atoms. For example, C1-6 alkyl refers to an alkyl group having ₁ to ₆ member atoms, eg, 1 to 4 member atoms. Alkyl groups can be straight or branched. Representative branched alkyl groups have one, two or three branches. Alkyl groups may be optionally substituted with one or more substituents as defined herein. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) base) and hexyl. _Alkyl groups can also be part of other groups such as _C1-6alkoxy .

同样地，术语“二甲基亚丁基”可以例如通过下列结构的任一种例举：

或

此外，术语“(C₁～C₆)亚烷基”意在包括此类支链烃基基团，诸如环丙基亚甲基，其可以通过下列结构例举：

"Alkylene" as used herein refers to a divalent saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms and in some embodiments 1 to 6 carbon atoms. "(C _a -C _b )alkylene" refers to an alkylene group having a to b carbon atoms. Alkylene groups include branched and straight chain hydrocarbyl groups. For example, "( _C1 - _C6 )alkylene" is intended to include methylene, ethylene, propylene, 2-methylpropylene, dimethylethylene, pentylene, and the like. Thus, the term "propylene" can be exemplified by the following structures:

Likewise, the term "dimethylbutylene" can be exemplified, for example, by any of the following structures:

or

Furthermore, the term "(C ₁ -C ₆ )alkylene" is intended to include such branched chain hydrocarbyl groups, such as cyclopropylmethylene, which can be exemplified by the following structures:

双环己基和

双环己基。金刚烷基包括但不限于以下结构：

In the present invention, "cycloalkyl" and "cycloalkane" refer to having multiple carbon atoms and no ring heteroatoms and having a single ring or multiple rings (including fused, bridged, spiro and adamantane systems ) saturated or partially saturated cyclic group. For polycyclic systems having aromatic and non-aromatic rings containing no ring heteroatoms, the term "cycloalkyl" applies when the point of attachment is at a non-aromatic carbon atom (eg 5,6,7,8,-tetra Hydronaphthalene-5-yl). The term "cycloalkyl" includes cycloalkenyl groups such as cyclohexenyl. Examples of cycloalkyl groups include, for example, adamantyl, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl. Examples of cycloalkyl groups including polybicycloalkyl ring systems are bicyclohexyl, bicyclopentyl, bicyclooctyl, and the like. Two such bicycloalkyl polycyclic structures are exemplified and named below:

dicyclohexyl and

Dicyclohexyl. Adamantyl includes but is not limited to the following structures:

"Alkenyl" means having 2 to 10 carbon atoms and in some embodiments 2 to 6 carbon atoms or 2 to 4 carbon atoms and having at least 1 site of vinyl unsaturation (>C=C<) straight or branched chain hydrocarbon groups. For example, (Ca-Cb)alkenyl refers to an alkenyl group having a to b carbon atoms and is intended to include, for example, vinyl, propenyl, isopropenyl, 1,3-butadienyl, and the like.

"Alkenylene" as used in the present invention refers to a hydrocarbon chain having 2 to 10 carbon atoms, at least one double bond and two unsaturated valences. For example, (C3 _{- C6} )alkenylene includes >C= _CH -CH2-, -CH-CH= _CH -CH2-, and the like.

"Alkynyl" means a straight-chain monovalent hydrocarbon group or a branched-chain monovalent hydrocarbon group containing at least one triple bond. The term "alkynyl" is also intended to include those hydrocarbyl groups having one triple bond and one double bond. For example, (C2-C6)alkynyl is intended to include ethynyl, propynyl, and the like.

The "alkynylene group" as used in the present invention refers to a divalent hydrocarbon chain having 2 to 10 carbon atoms and at least one triple bond.

"Halogen" is fluorine, chlorine, bromine or iodine.

"Haloalkyl", "halogen-substituted alkyl" means that the hydrogen atoms in the alkyl group may be replaced by one or more halogen atoms. For example, C _1-4 haloalkyl refers to an alkyl group containing 1 to 4 carbon atoms in which hydrogen atoms are replaced by one or more halogen atoms.

"-OR", "-NRR" and the like mentioned in the present invention mean that the R group is connected to an oxygen atom or a nitrogen atom by a single bond.

The oxygen atom in "-C(O)R", "-S(O) ₂ R" and the like described in the present invention is connected to a carbon atom or a sulfur atom by a double bond.

The "carbonyl group" in the present invention means that the oxygen atom is substituted by a double bond, that is, "=O".

硫代吗啉基、1，1-二氧代-硫代吗啉-4-基、氮杂环庚基、二氮杂环庚基、高哌嗪基或氧杂氮杂环庚基。二环饱和杂环烷基的实例是8-氮杂-二环[3.2.1]辛基、奎宁环基、8-氧杂-3-氮杂-二环[3.2.1]辛基、9-氮杂-二环[3.3.1]壬基、

部分不饱和杂环烷基的实例是二氢呋喃基、咪唑啉基、四氢-吡啶基或二氢吡喃基。In the present invention, "heterocycle", "heterocycloalkyl" and "heterocycloalkane" refer to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom; atoms, sulfur atoms, etc. Typically represents a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of multiple ring atoms, preferably a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of 3 to 9 ring atoms, comprising 1, 2 or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Bicyclic means consisting of two rings that share two ring atoms, i.e. the bridge separating the two rings is a single bond or a chain of one or two ring atoms. Examples of monocyclic saturated heterocycloalkyl are oxetanyl, azetidine, pyrrolidinyl, 2-oxo-pyrrolidin-3-yl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidine base, imidazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl,

Thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azepanyl, diazepanyl, homopiperazinyl or oxazepanyl. Examples of bicyclic saturated heterocycloalkyl groups are 8-aza-bicyclo[3.2.1]octyl, quinuclidinyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl, 9-aza-bicyclo[3.3.1]nonyl,

Examples of partially unsaturated heterocycloalkyl groups are dihydrofuranyl, imidazolinyl, tetrahydro-pyridyl or dihydropyranyl.

The "aromatic ring" and "aryl group" described in the present invention refer to an aromatic hydrocarbon group having a plurality of carbon atoms. Aryl groups are typically monocyclic, bicyclic or tricyclic aryl groups having 5 to 20 carbon atoms. Furthermore, the term "aryl" as used herein refers to an aromatic substituent which may be a single aromatic ring or multiple aromatic rings fused together. Non-limiting examples include phenyl, naphthyl or tetrahydronaphthyl.

In the present invention, "aromatic heterocycle" and "aromatic heterocyclic group" refer to an aromatic unsaturated ring containing at least one heteroatom; wherein the heteroatom refers to nitrogen atom, oxygen atom, sulfur atom and the like. Aromatic monocyclic or bicyclic hydrocarbons generally containing multiple ring atoms, one or more of which are heteroatoms selected from O, N, S. There are preferably one to three heteroatoms. Heterocyclic aryl groups represent, for example: pyridyl, indolyl, quinoxolinyl, quinolyl, isoquinolyl, benzothienyl, benzofuranyl, benzothienyl, benzopyranyl, benzene Thiapyranyl, furanyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, oxadiazolyl, benzimidazole base, benzothiazolyl, benzoxazolyl.

"Stereoisomer" includes enantiomers and diastereomers;

The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients that make up a pharmaceutical dosage form, and is physiologically Compatible with receptors.

The terms "salt" and "pharmaceutically acceptable salt" refer to the above-mentioned compounds or their stereoisomers, acid and/or base salts with inorganic and/or organic acids and bases, and also include zwitterionic salts ( inner salts), also including quaternary ammonium salts such as alkyl ammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. It can also be obtained by mixing the above-mentioned compound, or a stereoisomer thereof, with a certain amount of acid or base as appropriate (for example, an equivalent amount). These salts may be precipitated in solution and collected by filtration, recovered after evaporation of the solvent, or obtained by lyophilization after reaction in an aqueous medium. The salts described in the present invention can be hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate of the compound salt, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate.

Obviously, according to the above-mentioned content of the present invention, according to the common technical knowledge and conventional means in the field, without departing from the above-mentioned basic technical idea of the present invention, other various forms of modification, replacement or change can also be made.

Description of drawings

Figure 1 is a graph showing the results of fluorescence detection of the compounds of the present invention to form EGFR-PROTAC-CRBN/DDB1 ternary complexes.

Detailed ways

The above content of the present invention will be further described in detail below through the specific implementation in the form of examples. However, this should not be construed as limiting the scope of the above-mentioned subject matter of the present invention only to the following examples. All technologies implemented based on the above content of the present invention belong to the scope of the present invention.

The known starting materials of the present invention can be synthesized by using or according to methods known in the art, or can be purchased from companies such as Annagy Chemical, Chengdu Kelong Chemical, Shaoyuan Chemical Technology, Bailingwei Technology and the like.

The abbreviations of the reagents described in the examples are as follows: AIBN: azobisisobutyl cyanide; NBS: N-bromosuccinimide; DMP: DESS-MARTIN PERIODINANE Dess-Martin oxidant; DIPEA: N,N-diisobutylene Propylethylamine; TEMPO: 2,2,6,6-tetramethylpiperidine oxide; MTBE: methyl tert-butyl ether; NaHMDS: sodium bis(trimethylsilyl)amide; DIAD: azobis Diisopropyl formate; DPPA: Diphenylphosphoric azide; TBTU: 2-(1H-benzotriazo L-1-yl)-1,1,3,3-tetramethylurea tetrafluoroboric acid ester; TEA: triethylamine; SEMCl: 2-(trimethylsilyl)ethoxymethyl chloride; BINAP: 1,1'-binaphthyl-2,2'-bisdiphenylphosphine; X-Phos: 2 - Dicyclohexylphosphorus-2,4,6-triisopropylbiphenyl; DMF: N,N-dimethylformamide; DCM: dichloromethane; DCE: 1,2-dichloroethane; TFA: Trifluoroacetic acid; EA: ethyl acetate; PE: petroleum ether; BPO: benzoyl peroxide.

There is no special description in the examples, and the reaction is carried out under nitrogen atmosphere. There is no special description in the examples, and the solution refers to an aqueous solution. There is no special description in the examples, and the reaction temperature is room temperature. Room temperature is the most suitable reaction temperature, which is 20°C to 30°C. There is no special description in the examples, and M is moles per liter.

The structures of the compounds were determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR shifts ([delta]) are given in units of 10-6 (ppm). NMR was measured with (Bruker AvanceIII 400 and Bruker Avance 300) nuclear magnetic apparatus, and the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD) , the internal standard is tetramethylsilane (TMS). For the measurement of LC-MS, a Shimadzu LC-MS 2020 (ESI) was used. For the HPLC measurement, a Shimadzu high pressure liquid chromatograph (Shimadzu LC-20A) was used. MPLC (medium pressure preparative chromatography) used a Gilson GX-281 reverse phase preparative chromatograph. The thin layer chromatography silica gel plate is made of Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, and the specifications of the thin layer chromatography separation and purification products are 0.4mm ~ 0.5mm. Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

Synthesis of Intermediate Z-1

Step 1, Synthesis of 1-a

To a solution of methyl 2-methyl-3-bromobenzoate (51 g, 222.64 mmol) in 1,2-dichloroethane (560 mL) was added AIBN (3.66 g, 22.26 mmol) and NBS (47.55 g, 267.17 mmol) ). The reaction solution was heated to 85°C and reacted for 4 hours. After the reaction was completed, the reaction solution was concentrated to obtain 1-a (65 g, 211.06 mmol, 94.80% yield, crude product), which was used in the next step without purification. MS(ESI) m/z=306[M+1] ⁺ .

Step 2. Synthesis of 1-b

To a solution of 1-a (42 g, 136.38 mmol) and 3-aminopiperidine-2,6-dione hydrochloride (17.47 g, 136.38 mmol) in acetonitrile (210 mL) was added DIPEA (21.15 g, 163.65 mmol, 28.51 mmol) mL), the reaction solution was heated to 90° C. and stirred at this temperature for 3 hours. After the reaction was completed, the mixture was concentrated to obtain a crude product, which was washed with water and filtered to obtain compound 1-b (35.6 g, 110.17 mmol, 80.78% yield). MS(ESI) m/z=324[M+1] ⁺ .

Step 3. Synthesis of 1-c

To a solution of 1-b (3 g, 9.28 mmol) in DMF (30 mL) were sequentially added 4-pentyn-1-ol (1.56 g, 13.93 mmol), cuprous iodide (265.22 mg, 1.39 mmol), Pd (Ph ₃ P) ₂ Cl ₂ (488.10 mg, 696.29 μmol) and DIPEA (2.82 g, 27.85 mmol, 3.88 mL). The reaction solution was stirred at 80°C for 18 hours under the protection of N ₂ , diluted with water, extracted with ethyl acetate, and the separated organic phase was washed with water and brine, dried and then spin-dried. The crude product was added with ethyl acetate, and a brown solid was precipitated. After filtration, the filter cake was collected to obtain the target compound 1-c (1.6 g, 4.51 mmol, 48.65% yield). MS(ESI) m/z=355[M+1] ⁺ .

Step 4. Synthesis of 1-d

To a mixed solution of 1-c (200 mg, 0.61 mmol) in MeOH (80 mL)/THF (20 mL)/EtOAc (20 mL) was added Pd/C (200 mg). After the mixture was replaced with hydrogen, it was stirred at room temperature for 18 hours under the protection of hydrogen. After the reaction was completed, Pd/C was removed by filtration, and the filtrate was concentrated to give 1-d (162 mg, 0.49 mmol, 80% yield, crude).

Step 5. Synthesis of Intermediate Z-1

To a mixed solution of compound 1-d (300 mg, 881.39 μmol) in DCM (3 mL) and DMF (3 mL) was added Dess Martin’s reagent (560.75 mg, 1.32 mmol). The suspension was stirred at room temperature for 1 hour. After the reaction was completed, the mixed solution was filtered, the filtrate was concentrated and spin-dried, the crude product was diluted with ethyl acetate, a brown solid was precipitated, the solid was collected by filtration, and separated and purified with a reverse-phase C18 column to obtain the intermediate compound Z-1 (200 mg, 591.09 μmol, 67.06% yield). MS(ESI) m/z=339[M+1] ⁺ .

Synthesis of Intermediate Z-2

According to step 2 to step 5 in the synthesis method of intermediate Z-1, the methyl 2-bromomethyl-4-bromobenzoate used in step 1 is used as the raw material, and other conditions remain unchanged to obtain Z-2. MS(ESI) m/z=339[M+1] ⁺ .

Synthesis of Intermediate Z-3

According to steps 1 to 5 in the synthetic method of intermediate Z-1, the methyl 2-methyl-3-bromobenzoate in step 1 is replaced with methyl 2-methyl-5-bromobenzoate, and the rest With the conditions unchanged, Z-3 is obtained. MS(ESI) m/z=339[M+1] ⁺ .

Synthesis of Intermediates Z-4 to Z-12

According to step 1 to step 5 in the synthetic method of intermediate Z-1, respectively replace methyl 2-methyl-3-bromobenzoate in step 1 with SM-Br in the table, and replace 4- The pentyn-1-ol was replaced with the corresponding SM-ene in the table, and the other conditions remained unchanged to obtain the corresponding structure of Z-4 and Z-12 in the table.

Synthesis of Intermediate Z-13

At zero temperature, DMP (974.75 mg, 2.30 mmol) was added to a mixed solution of intermediate compound 1-c (500 mg, 1.53 mmol) in DMF (4 mL)/DCM (40 mL), and the reaction mixture was slowly warmed to room temperature and stirred for 3 hours After the reaction was completed, 1 mL of methanol was added, the reaction mixture was spin-dried, and the residue was separated and purified by m-HPLC to obtain the intermediate compound Z-13 (133 mg, 410.08 μmol, 26.77% yield). MS(ESI) m/z=325[M+1] ⁺ .

Synthesis of Intermediate Z-14

Referring to the synthesis method of compound Z-13, the raw material 1-c is replaced by 2-c, and the other methods remain unchanged to obtain Z-14. MS(ESI) m/z=325[M+1] ⁺ .

Synthesis of Intermediate Z-15

Referring to the synthesis method of compound Z-13, the raw material 1-c is replaced by 3-c, and the other methods remain unchanged to obtain Z-15. MS(ESI) m/z=325[M+1] ⁺ .

Synthesis of Intermediates Z-16 to Z-27

According to the synthesis method of the intermediate Z-13, the raw material 1-c in it is replaced with the corresponding raw material intermediate number, and the other methods remain unchanged to obtain the corresponding Z-16 to Z-27 in the table.

Synthesis of Intermediate Z-25

To a mixed solution of 1-c (1 g, 3.06 mmol) in acetonitrile (66.67 mL)/water (133.33 mL) were added TEMPO (191.51 mg, 1.23 mmol) and iodobenzenediacetic acid (3.95 g, 12.26 mmol). The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction solution was concentrated, and the crude product was separated and purified by a normal phase silica gel column (DCM:MEOH=33:1) to obtain the intermediate compound Z-25 (248 mg, 728.71 μmol, 23.78% yield). ). MS(ESI) m/z=341[M+1] ⁺ .

Synthesis of Intermediate Z-26

To a solution of intermediate Z-25 (160 mg, 0.47 mmol) in MeOH (10 mL) was added Pd/C (30 mg). The reaction solution was replaced with hydrogen, and then stirred at room temperature for 1 hour under the protection of hydrogen. After the reaction was completed, the Pd/C was filtered off, and the filtrate was spin-dried to obtain the intermediate compound Z-26 (150 mg, 93% yield). MS(ESI)m/z=345[M+1] ⁺

Synthesis of Intermediate Z-27

Step 1, Synthesis of 27-a

To a solution of delta-valerolactone (10 g, 99.88 mmol) in toluene (100 mL) was added BnBr (68.32 g, 399.54 mmol) and KOH (16.81 g, 299.65 mmol) in water (100 mL), and the mixture was heated to 115 °C for reaction overnight. After the reaction was completed, it was cooled to room temperature, 7g of KOH was added, the liquid was separated, the separated aqueous phase was washed twice with MTBE, then adjusted to pH=2-3 with concentrated hydrochloric acid, extracted with ethyl acetate, and the obtained organic phase was dried and concentrated. Obtained intermediate compound 27-a (5.4 g, 25.93 mmol, 25.96% yield), MS (ESI) m/z=207[M+1] ⁺

Step 2. Synthesis of 27-b

At zero degrees, 1 drop of DMF was added to a solution of 27-a (2 g, 9.60 mmol) in DCM (100 mL), then (COCl) ₂ (2.44 g, 19.21 mmol) was slowly added dropwise, and the reaction was completed at room temperature for one hour. After that, the reaction solution was concentrated to obtain compound 27-b (2.1 g, 9.26 mmol, 96.46% yield).

Step 3. Synthesis of 27-c

(R)-4-benzyl-2-oxazolidinone (5.77g, 32.55mmol) was dissolved in THF (80mL), the temperature was lowered to -78°C under nitrogen protection, n-BuLi (16mL) was added, and the reaction was incubated After 30 minutes, the THF solution of 27-b (7.38 g, 32.55 mmol) was added, the dropping was completed, and the reaction was incubated for 1 hour. After the reaction was completed, the reaction was poured into a saturated aqueous ammonium chloride solution to quench, and the liquids were separated.

The organic phase was concentrated to dryness, and the residue was separated and purified by normal phase silica gel column (PE/EA=3/1) to obtain 27-c (7.04 g, 19.16 mmol, 58.85% yield). MS(ESI)m/z=368[M+1] ⁺

Step 4. Synthesis of 27-d

-78°C, under nitrogen protection, to 27-c (7.04g, 19.16mmol) in THF (50mL), NaHMDS (12.5mL) was added dropwise, the reaction was incubated for 30 minutes, and then MeI (16.32g, 114.96mmol) was added dropwise ) in THF solution, the dripping was completed, the external bath was removed, and the reaction was carried out at room temperature for 1 hour. After the reaction was completed, the reaction solution was poured into a saturated aqueous ammonium chloride solution to quench, the layers were separated, the organic phase was concentrated, and the residue was separated by a normal-phase silica gel column. Purification (MTBE/PE=1/1) gave 27-d (5.4 g, 14.16 mmol, 73.88% yield). MS(ESI)m/z=382[M+1] ⁺

Step 5. Synthesis of 27-e

At zero temperature, LiAlH ₄ (15 mL) was added to 27-d (5.4 g, 14.16 mmol) in THF (100 mL), and the reaction was raised to room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated to dryness, and the residue was separated and purified by normal phase silica gel column (MTBE/PE=1/2) to obtain 27-e (2.4 g, 11.52 mmol, 81.39% yield), MS (ESI) m /z=209[M+1] ⁺

Step 6, Synthesis of 27-f

27-e (2.4 g, 11.52 mmol) was dissolved in THF (100 mL), PPh ₃ (6.04 g, 23.04 mmol), DIAD (4.65 g, 23.04 mmol) and DPPA (6.34 g, 23.04 mmol) were added successively under nitrogen protection ), stirred at room temperature overnight, after completion of the reaction, the reaction solution was concentrated to dryness, and the residue was separated and purified by normal phase silica gel column (8% MTBE in PE) to obtain 27-f (1.97 g, 8.44 mmol, 73.28% yield). MS(ESI)m/z=234[M+1] ⁺

Step 7, Synthesis of 27-g

To 27-f (2 g, 8.57 mmol) in MeOH (50 mL) was added 2 drops of concentrated hydrochloric acid followed by Pd/C (0.5 g). After hydrogen substitution, the mixture was stirred at room temperature for 2 hours. After the reaction was completed, Pd/C was filtered off, and the filtrate was concentrated to dryness to obtain 27-g (1 g, 8.53 mmol, 99.54% yield). MS(ESI)m/z=118[M+1] ⁺

Step 8, Synthesis of 27-h

To a solution of 27-g (1 g, 8.53 mmol) in DMF (20 mL) was added K ₂ CO ₃ (2.36 g, 17.07 mmol) followed by 2-fluoro-4-bromonitrobenzene (1.88 g, 8.53 mmol), The temperature was raised to 60°C and stirred overnight. After the reaction was completed, the reaction solution was concentrated, and the residue was separated and purified by normal phase silica gel column (PE/EA=5/1) to obtain 27-h (2 g, 6.31 mmol, 73.89% yield). MS(ESI)m/z=318[M+1] ⁺

Step 9, Synthesis of 27-i

At zero temperature, to a solution of 27-h (465 mg, 1.47 mmol) in DCM (10 mL), TEA (222.52 mg, 2.20 mmol, 306.72 μL) and MsCl (201.52 mg, 1.76 mmol) were added in sequence, and the mixture was stirred at zero temperature for 1 hour, After the completion of the reaction, saturated aqueous sodium bicarbonate solution was added to separate the layers. The separated organic phase was washed with water and saturated brine successively, dried over anhydrous sodium sulfate, and concentrated to obtain 27-i (0.57 g, 1.44 mmol, 98.36% yield), the crude product was directly used in the next reaction without purification. MS(ESI)m/z=397[M+1] ⁺

Synthesis of Steps 10, 27-j

27-i (579 mg, 1.46 mmol), methyl 2-(5-hydroxy-1-methyl-1H-pyrazole-4-yl)-6-methylisonicotinate (362.18 mg, 1.46 mmol) and K ₂ CO ₃ (505.37 mg, 3.66 mmol) were dissolved in DMF (15 mL), the reaction was raised to 60 ° C and the reaction was stirred overnight. After the reaction was completed, it was cooled to room temperature, MTBE was added, the mixture was washed twice with water, and the separated organic The phases were concentrated and the residue was purified by silica gel column separation (60% EA in PE) to give 27-j (254 mg, 464.85 μmol, 31.73% yield). MS(ESI)m/z=548[M+1] ⁺

Step 11, Synthesis of 27-k

To 27-j (254 mg, 464.85 μmol) in MeOH (10 mL) at zero temperature, NH ₄ OH (3 mL) and Na ₂ S ₂ O ₄ (404.42 mg, 2.32 mmol) were sequentially added, and the reaction was completed by stirring at room temperature for 1 hour. Then, washed with water and extracted with ethyl acetate. The combined organic phases were washed with water and saturated brine successively, dried, and concentrated to obtain 27-k (170 mg, 329.18 μmol, 70.81% yield). The crude product was not purified and was directly used for the next reaction. MS(ESI)m/z=518[M+1] ⁺

Synthesis of Step 12, 27-1

To a mixed solution of 27-k (170 mg, 329.18 μmol) in DCM (10 mL)/t-BuOH (3 mL), CNBr (41.87 mg, 395.02 μmol) was added, and the mixture was stirred at room temperature overnight. After the reaction was completed, a saturated aqueous sodium bicarbonate solution was added, stirred for 10 minutes, and the layers were separated. The organic phase was washed with a saturated aqueous sodium bicarbonate solution, water, and brine, dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain 27-1 (177 mg , 326.91 μmol, 99.31% yield). MS(ESI)m/z=541[M+1] ⁺

Step 13, Synthesis of 27-m

To 27-1 (177 mg, 326.91 μmol) of THF (5 mL)/H ₂ O (5 mL), NaOH (52.31 mg, 1.31 mmol) was added, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated and diluted with water. The pH was adjusted to 4-5 with 1M HCl, a solid was precipitated, filtered, and the filter cake was washed with water, and dried to obtain 27-m (172 mg, 326.12 μmol, 99.76% yield). MS(ESI)m/z=541[M+1] ⁺

Step 14. Synthesis of Z-27

To 27-m (172 mg, 326.12 μmol) in DCM (5 mL), TEA (132.00 mg, 1.30 mmol, 181.94 uL) and TBTU (100.14 mg, 391.34 μmol) were sequentially added, and stirred at room temperature for 15 minutes. After the reaction was completed, water was added. The organic phase was diluted, separated, and the organic phase was concentrated, and the obtained residue was separated and purified by silica gel column (DCM/MeOH=10/1) to obtain intermediate Z-27 (113 mg, 221.83 μmol, 68.02% yield). MS(ESI) m/z=511[M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 12.83(s, 1H), 8.49-8.33(m, 1H), 7.92(d, J =1.4Hz,2H),7.56(d,J=1.2Hz,1H),7.46(d,J=8.4Hz,1H),7.38(dd,J=8.5,1.8Hz,1H),4.35(dt,J =9.7,4.9Hz,1H),4.14(dd,J=13.7,3.2Hz,1H),4.04–3.90(m,2H),3.73(s,3H),2.79(s,1H),2.69(s, 1H), 2.55(s, 3H), 2.28–2.13(m, 1H), 1.95(d, J=14.5Hz, 1H), 1.22(s, 1H), 0.81(d, J=6.5Hz, 3H).

Synthesis of Intermediate Z-28

Step 1, Synthesis of 28-a

To a solution of intermediate 27 (493.00 mg, 967.81 μmol) in DMF (30 mL) was added DIPEA (375.24 mg, 2.90 mmol, 505.71 μL) and SEMCl (242.44 mg, 1.45 mmol), and the reaction mixture was warmed to 60 °C and stirred overnight , after the completion of the reaction, the reaction solution was concentrated, the residue was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine successively, dried over anhydrous sodium sulfate, and after concentration, the residue was separated and purified with a normal phase silica gel column (100% EA) to give 28-a (373 mg, 583.13 μmol, 60.25% yield). MS(ESI) m/z=639[M+1] ⁺ .

Step 2. Synthesis of 28-b

To a mixed solution of 28-a (718 mg, 1.12 mmol) in dioxane (20 mL)/water (2 mL) was added 4-N-Boc piperazine 1-methylboron trifluoride potassium salt (515.51 mg, 1.68 g mmol), X-Phos (106.86 mg, 224.50 μmol), Pd(OAc) ₂ (25.14 mg, 112.25 μmol) and Cs ₂ CO ₃ (1.10 g, 3.37 mmol), the mixture was replaced with nitrogen several times, and then heated under nitrogen The temperature was raised to 100 °C under protection and stirred for 15 h. After the reaction was completed, the reaction solution was concentrated, the residue was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, and dried over anhydrous sodium sulfate. After concentration, the residue was separated and purified by normal phase silica gel column (100% EA). ) to give 28-b (607 mg, 799.71 μmol, 71.25% yield). MS(ESI) m/z=759[M+1] ⁺ , ¹ HNMR (400MHz, DMSO-d ₆ ) δ 12.83(s, 1H), 8.49-8.33(m, 1H), 7.92(d, J= 1.4Hz, 2H), 7.56(d, J=1.2Hz, 1H), 7.46(d, J=8.4Hz, 1H), 7.38(dd, J=8.5, 1.8Hz, 1H), 4.35(m, 1H) ,4.14(dd,J＝13.7,3.2Hz,1H),4.04–3.90(m,2H),3.73(s,3H),2.79(s,1H),2.69(s,1H),2.55(s,3H) ), 2.28-2.13(m, 1H), 1.95(d, J=14.5Hz, 1H), 1.43-1.47(m, 1H), 1.22(s, 1H), 0.81(d, J=6.5Hz, 3H) .

Step 3. Synthesis of Z-28

To 28-b (671 mg, 884.03 μmol) in DCM (5 mL), TFA (5 mL) was added, and the reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, the dry reaction solution was concentrated to obtain Z-28 (452 mg, 855.01 μmol, 96.72% yield), which was directly used in the next reaction without purification. MS(ESI) m/z=529[M+1] ⁺ .

Synthesis of Intermediate Z-29

Referring to step 8 to step 14 in the synthesis method of Z-27, only in step 8, 2-fluoro-5-bromonitrobenzene is used instead of 2-fluoro-4-bromonitrobenzene, and the rest of the methods are the same, and the obtained Z-29. MS(ESI) m/z=509[M+1] ⁺ .

Synthesis of Intermediate Z-30

Referring to the methods from step 1 to step 3 of the synthesis method of Z-28, Z-29 is used to replace Z-27 in step 1, and the remaining method steps are the same to obtain Z-30. MS(ESI) m/z=529[M+1] ⁺ .

Synthesis of Intermediate Z-31

Step 1. Synthesis of Intermediate 31-a

To a solution of Z-27 (493.00 mg, 967.81 μmol) in DMF (30 mL), DIPEA (375.24 mg, 2.90 mmol, 505.71 μL) and SEMCl (242.44 mg, 1.45 mmol) were sequentially added, and the reaction mixture was heated to 60° C. and stirred 12h, after the completion of the reaction, the reaction solution was concentrated, the residue was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine successively, dried over anhydrous sodium sulfate, and then spin-dried and separated and purified with a normal phase silica gel column ( 100% ethyl acetate) to give 31-a (373 mg, 583.13 μmol, 60.25% yield). MS(ESI) m/z=639[M+1] ⁺ .

Step 2. Synthesis of Intermediate 31-b

To a solution of 31-a (50 mg, 78.17 μmol) in toluene (3 mL) was added 1-tert-butoxycarbonylpiperazine (14.56 mg, 78.17 μmol), BINAP (9.73 mg, 15.63 μmol), Pd(OAc) ₂ (1.75 mg, 7.82 μmol) and Cs ₂ CO ₃ (50.81 mg, 156.33 μmol). Under nitrogen, the temperature was raised to 120°C and the reaction was stirred at this temperature for 15 hours. After the reaction was completed, the mixture was diluted with ethyl acetate, washed with water, and the organic phase was concentrated to obtain a crude product. The crude product was separated and purified by a reverse-phase silica gel column to obtain 31-b (30 mg, 40.27 μmol, 51.52% yield). MS (ESI) m /z=745[M+1] ⁺ .

Step 3. Synthesis of Intermediate Z-31

To a solution of 31-b (30 mg, 40.27 μmol) in DCM (3 mL) was added TFA (2 mL), and the mixture was stirred at room temperature for 1 hour. Then, the reaction solution was concentrated, and the residual TFA was spun off with an oil pump to obtain Z-31 (50 mg, 43.8% purity), which was directly used in the next reaction without purification. MS(ESI) m/z=515[M+1] ⁺ .

Synthesis of Intermediate Z-32

Referring to steps 1 to 3 in the synthesis method of Z-31, Z-29 was used to replace Z-27 in step 1, and Z-32 was obtained by the same method. MS(ESI) m/z=515[M+1] ⁺ .

Synthesis of Intermediate Z-33

Step 1. Synthesis of Intermediate 33-a

To a solution of 4-bromo-2-methylaminoaniline (7.6 g, 37.8 mmol) in THF (200 mL) was added CDI (7.4 g, 45.6 mmol) at room temperature. The mixture was refluxed for 16 hours, the reaction solution was cooled to room temperature, diluted with water (500 mL), and stirred at room temperature for 30 minutes. The resulting precipitate was collected by filtration, washed with water, and the filter cake was dried to give 6-bromo-1-methyl-1,3-dihydro-2H-benzimidazol-2-one, namely 33-a (7 g, 72 % yield) yellow solid, MS (ESI) m/z = 227.10, 229.10 [M+1] ⁺ .

Step 2. Synthesis of Intermediate 33-b

To 3-bromopiperidine-2,6-dione (10 g, 52.08 mmol), 4-methoxybenzyl alcohol (7.92 g, 57.29 mmol) and _PPh3 (13.66 g, 52.08 mmol) under an ice bath under nitrogen ) was added dropwise to the mixed solution of DEAD (9.06g, 52.08mmol), the reaction mixture was stirred at zero degrees for 2 hours, after the reaction was completed, water was added to quench the reaction, the mixed solution was extracted with ethyl acetate, and the combined organic phase was concentrated to obtain the crude product , separated and purified by silica gel column (PE/EA=3/1 and DCM/PE=1/1, v/v) to obtain intermediate 33-b (4.88 g, 15.63 mmol, 30.02% yield). MS(ESI) m/z=312,314[M+1] ⁺ .

Step 3. Synthesis of Intermediate 33-c

The solution of 33-a (4.0 g, 17.7 mmol) in THF (50 mL) was cooled to 0 °C, t-BuOK (2.38 g, 21.2 mmol) was added, the mixture was stirred at zero temperature for 30 minutes, and then 33-b (8.26 g) was added. , 26.5 mmol), the mixture was stirred at room temperature overnight, the solvent was removed, and the crude product was separated and purified by silica gel column (EtOAc:DCM=1:1) to obtain 33-c (2.9 g, 6.33 mmol, yield: 36%), MS ( ESI)m/z=458[M+1] ⁺ .

Step 4. Synthesis of Intermediate Z-33

To a solution of 33-c (2.0 g, 4.37 mmol) in toluene (20 mL) was added _MeSO3H (10 mL) at room temperature, and the reaction mixture was warmed to 110 °C and stirred at this temperature for 2 h. Cooled to room temperature, the mixture was concentrated under reduced pressure, the crude product was diluted with ice water, the precipitated solid was collected by filtration, washed with n-hexane, and dried to obtain intermediate Z-33 (0.8 g, yield 54%), MS (ESI) m/z=339[M+1] ⁺ .

Synthesis of Intermediate Z-34

Step 1. Synthesis of Intermediate 34-a

Referring to the synthetic method of 33-a in the step 1 of the intermediate Z-33, the raw material 4-bromo-2-methylaminoaniline is replaced with 5-bromo-2-methylaminoaniline, and the remaining operations are unchanged, and the intermediate can be obtained 34-a. MS(ESI) m/z = 227.10, 229.10 [M+1] ⁺ .

Step 2. Synthesis of Intermediate 34-b

To tetrahydro-5-oxo-2-furancarboxylic acid (12 g, 92 mmol) was slowly added SOCl ₂ (25 g, 207 mmol) at zero degrees. The mixture was heated to 85°C and stirred for 3 hours, then the reaction solution was cooled to room temperature and stirred for 6 hours. After the reaction of the raw materials was completed, the mixture was concentrated under reduced pressure. Under nitrogen protection, the crude product was dissolved in zero-degree dry DCM, then Et ₃ N (18.7 g, 184 mmol) and 4-methoxybenzylamine (10.1 g, 74 mmol) were added, and the mixture was stirred at room temperature for 3 hours. After completion, diluted with water, the mixture was extracted with DCM, the combined organic phases were washed with 0.5M aqueous HCl (50 mL) and saturated brine (50 mL) successively, dried over anhydrous sodium sulfate, filtered and spin-dried, and the crude product was separated with a silica gel column Purification gave 34-b (10.5 g, 45.6% yield), MS (ESI) m/z = 272.0 [M+Na] ⁺ .

Step 3. Synthesis of Intermediate 34-c

34-b (10.5 g, 47.9 mmol) in anhydrous THF (120 mL) was cooled to -78 °C, then under nitrogen protection and at -78 °C, t-BuOK (5.4 g, 48.4 mmol) in anhydrous THF ( 100 mL), the reaction mixture was stirred at -40 °C for 1 hour, after the reaction was completed, the mixture was quenched with saturated NH ₄ Cl solution, extracted with ethyl acetate, the combined organic phases were washed with saturated brine, and anhydrous sodium sulfate was added. After drying, filtration and concentration, the vinegar flask was separated and purified by silica gel column to obtain intermediate 34-c (10.9 g, 43.73 mmol, 77.7% yield). MS(ESI) m/z=250.0[M+1] ⁺ .

Step 4. Synthesis of intermediate 34-d

Trifluoromethanesulfonic anhydride (18.51 g, 65.59 mmol) was added dropwise to a solution of 34-c (10.9 g, 43.73 mmol) and pyridine (6.92 g, 87.46 mmol) in DCM (100 mL) at zero temperature, under nitrogen protection, The mixture was stirred at 0-10°C for 1.5 hours. After the reaction was completed, the mixture was concentrated, and the crude product was separated and purified by silica gel column to obtain 34-d (8.17 g, 21.43 mmol, 49% yield), MS (ESI) m/z=382 [M+1] ⁺ .

Step 5. Synthesis of Intermediate 34-e

At zero degrees, t-BuOK (2.89 g, 25.7 mmol) was added to a solution of 34-a (4.86 g, 21.43 mmol) in THF (100 mL), the mixture was stirred at 0 °C for 0.5 h, and then 34-d was added dropwise (8.17 g, 21.43 mmol) in THF (50 mL) solution, the reaction solution was stirred at room temperature for 0.5 h under nitrogen protection. After the reaction was completed, the reaction solution was quenched with saturated NH ₄ Cl (40 mL), extracted with ethyl acetate, the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was separated and purified by mHPLC to obtain 34-e (4.91 g, 10.71 mmol, 55% yield). MS(ESI) m/z=459[M+1] ⁺ .

Step 6. Synthesis of Intermediate Z-34

Referring to step 4 of intermediate Z-33, replace the substrate 33-c with 34-e in the step, and the rest of the operation steps are the same to obtain intermediate Z-34, MS(ESI) m/z=338,340[M+ 1] ⁺ .

Synthesis of Intermediate Z-35

Referring to the synthetic route of intermediate Z-33, the method from step 1 to step 4, in step 1, 3-bromo-2-methylaminoaniline is used instead of 4-bromo-2-methylaminoaniline as the starting material, The remaining operations are the same and go through the same steps to obtain the intermediate Z-35, MS(ESI) m/z=338,340[M+1] ⁺ .

Synthesis of Intermediate Z-36

Step 1. Synthesis of Intermediate 36-a

4-Pyrazole boronic acid pinacol ester (5 g, 25.77 mmol) and 3-benzyloxybromopropane (8.86 g, 38.65 mmol) were mixed in DMF (20 mL) solution, then K ₂ CO ₃ (8.90 g, 64.42 mmol), KI (427.76 mg, 2.58 mmol). The reaction mixture was warmed to 80°C and stirred for 17 hours. After the reaction was completed, the crude product obtained by concentrating the reaction solution was separated and purified by silica gel column (PE:EA=8:1, v/v) to obtain intermediate 36-a (6.4 g, 18.70 mmol, 72.57% yield), MS (ESI) m/z=343 [M+H] ⁺ .

Step 2. Synthesis of Intermediate 36-b

36-a (2 g, 5.84 mmol) and 1-b (1.89 g, 5.84 mmol) were mixed in DMF (20 mL), Na ₂ CO ₃ (1.86 g, 17.53 mmol) and Pd(dppf)Cl ₂ (213.80 were added) mg, 292.19 umol), the mixture was replaced with nitrogen several times, then heated to 100° C. under the protection of nitrogen, and stirred overnight. After the reaction was completed, it was cooled to room temperature, diluted with water, extracted with ethyl acetate, the combined organic phases were washed with saturated brine, concentrated, and the crude product was separated and purified by silica gel column (100% EA) to obtain intermediate 36-b (1.6 g). , 3.49 mmol, 59.71% yield), MS (ESI) m/z=459 [M+H] ⁺ .

Step 3. Synthesis of Intermediate 36-c

To a solution of 36-b (840 mg, 1.83 mmol) in MeOH (100 mL) was added 10% Pd/C (840 mg, 1.83 mmol), the mixture was replaced with hydrogen, and the mixture was stirred at room temperature for 24 hours under the protection of hydrogen. After the reaction was completed, Pd/C was filtered off, and the filtrate was concentrated to obtain 36-c (367 mg, 996.24 μmol, 54.38% yield). MS (ESI) m/z = 369 [M+H] ⁺ .

Step 4. Synthesis of Intermediate Z-36

To a solution of 36-c (100 mg, 271.45 μmol) in DCM (30 mL) was added DMP (230.19 mg, 542.91 μmol) and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and the obtained crude product was separated and purified by silica gel column (DCM/MeOH=10/1) to obtain Z-36 (85 mg, 232.01 μmol, 85.47% yield). MS (ESI) m/z = 367 [M+H] ⁺ .

Synthesis of Intermediate Z-37

Step 1. Synthesis of intermediate 37-a

To pinacol 4-pyrazoleboronic acid (6.0 g, 30.92 mmol) in acetonitrile (60 ml) was added CsF (470 mg, 3.09 mmol) and tert-butyl acrylate (5.82 mL, 40.2 mmol). The reaction mixture was warmed to 80°C and stirred at this temperature overnight. After the reaction was completed, it was cooled to room temperature, concentrated under reduced pressure, and the crude product was extracted with water and ethyl acetate. The separated organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The obtained crude product was passed through a silica gel column. Separation and purification (n-hexane/ethyl acetate=1/1, v/v) gave 37-a (9.37 g, 29.07 mmol, yield 94%). MS (ESI) m/z = 323 [M+H] ⁺ .

Step 2. Synthesis of Intermediate 37-b

To a solution of 37-a (500 mg, 1.55 mmol) and 1-b (455.86 mg, 1.41 mmol) in DMF (12 mL)/H ₂ O (3 mL) was added K ₃ PO ₄ (416.73 mg, 1.98 mmol), S- phos (69.41 mg, 169.29 μmol) and Pd(OAc) ₂ (22.61 mg, 98.75 μmol), the mixture was replaced with nitrogen and then heated to 100° C. under the protection of nitrogen, and stirred at this temperature for 15 hours, after the reaction was completed. Cooled to room temperature, filtered, concentrated the filtrate, the crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, concentrated, and the crude product was separated and purified by silica gel column (PE/EA=0 /1, v/v) to give 37-b (100 mg, 228.06 μmol, 16.17% yield) as a pale yellow solid. MS (ESI) m/z = 439 [M+H] ⁺ .

Step 3. Synthesis of Intermediate Z-37

To a solution of 37-b (100 mg, 228.06 μmol) in DCM (2 mL) was added TFA (1.54 g, 13.51 mmol, 1 mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated and dried to obtain Z-37 (92 mg, 185.34 μmol, 81.27% yield), which was directly used in the next reaction without purification. MS (ESI) m/z = 383 [M+H] ⁺ .

Synthesis of Intermediate Z-38

Step 1. Synthesis of Intermediate 38-a

To a solution of methyl 2-methyl 5-iodobenzoate (10 g, 36.22 mmol) in CCl ₄ (100 mL) was added NBS (6.77 g, 38.03 mmol) and benzoyl peroxide (4.38 g, 18.11 mmol), and mixed The temperature of the liquid was raised to 80°C and the reaction was stirred for 4 hours. After the reaction was completed, it was cooled to room temperature, filtered, and the filtrate was concentrated to obtain 38-a (12 g, 33.81 mmol, 93.33% yield) as an orange oil, which was used in the next reaction without further purification.

Step 2. Synthesis of Intermediate 38-b

To a solution of 38-a (12 g, 33.81 mmol) in DMF (150 mL) was added ₃ -amino-2,6-piperidinedione hydrochloride (6.12 g, 37.19 mmol) and K2CO3 (14.02 _g , 101.42 mmol). The reaction mixture was heated to 70 °C and stirred for 18 h. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water, stirred for 10 min, filtered, the filter cake was washed with petroleum ether and ethyl acetate, the filter cake was collected, and dried to obtain 38-b (6.5 g, 17.56 mmol, 51.95% yield). ). MS (ESI) m/z=371 [M+H] ⁺ .

Step 3. Synthesis of Intermediate 38-c

To 38-b (400 mg, 1.08 mmol) and 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboropentan-2-yl)-1H-pyrazole -1-yl) tert-butyl acetate (366.34 mg, 1.19 mmol) in dioxane (5 mL)/H2O (0.5 mL) was added K ₃ PO ₄ (320.74 mg, 1.51 mmol), S-phos (53.17 mg, 129.68 μmol) and Pd(OAc) ₂ (17.32 mg, 75.65 μmol), the mixture was replaced with nitrogen several times, then heated to 100° C. under nitrogen protection, and stirred overnight. After the reaction was completed, it was cooled to room temperature, diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and the concentrated crude product was recrystallized from a mixture of DMF/H ₂ O 38-c (200 mg, 471.20 μmol, 43.60% yield) was obtained as a pale yellow solid. MS (ESI) m/z = 425 [M+H] ⁺ .

Step 4. Synthesis of Intermediate Z-38

To a solution of 38-c (200 mg, 471.20 μmol) in DCM (3 mL) was added TFA (53.73 mg, 471.20 μmol, 35.12 μL) at room temperature, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated, and the crude product was dried to obtain Z-38 (200 mg, 414.62 μmol, 87.99% yield, TF), which was directly used in the next reaction without further purification.

Synthesis of Intermediate Z-39

Step 1. Synthesis of Intermediate 39-a

To a mixture of intermediates 38-b (1 g, 2.70 mmol) and 36-a (1.02 g, 2.97 mmol) in dioxane (20 mL)/H ₂ O (1 mL) was added Na ₂ CO ₃ (400.93 mg) , 3.78mmol), S-phos (132.92mg, 324.20μmol) and Pd(OAc) ₂ (43.31mg, 189.12μmol), the mixture was replaced with nitrogen several times, and the reaction mixture was warmed to 60° C. under the protection of nitrogen. The reaction was stirred at 60°C overnight. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, the obtained crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and spin-dried to obtain the crude product by pre-HPLC separation and purification to obtain Intermediate 39-a (130 mg, 283.53 μmol, 10.49% yield) as a white solid. MS(ESI) m/z=284[M+H] ⁺ .

Step 2. Synthesis of Intermediate 39-b

Pd/C (1.09 g, 894.20 μmol, 10% purity) was added to a mixture of 39-a (410 mg, 894.20 μmol) in MeOH (150 mL) and THF (30 mL), the reaction solution was replaced with hydrogen several times, and then added under hydrogen The mixture was stirred at room temperature for 15 h under protection, filtered, and the filtrate was concentrated and dried to obtain the crude product (310 mg) of intermediate 39-b, which was directly used in the next reaction without purification.

Step 3. Synthesis of Intermediate Z-39

DMP (345 mg, 0.81 mmol) was added to a mixture of 39-b (150 mg, 0.41 mmol) in DCM (3 mL)/DMF (1 mL) at room temperature, and the reaction mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and the obtained crude product was separated and purified by silica gel column to obtain Z-39 (110 mg, 0.3 mmol, 73% yield). MS (ESI) m/z = 367 [M+H] ⁺ .

Synthesis of Intermediate Z-40

Step 1. Synthesis of Intermediate 40-a

Under ice bath, SEMCl (9.9 mL, 56 mmol) was added to a solution of 2-b (10.0 g, 30.9 mmol) and DBU (10.4 mL, 69 mmol) in DMF (95 mL), the mixture was slowly warmed to room temperature and the reaction was stirred for 6 h. After the completion of the reaction, the reaction solution was quenched with saturated NH ₄ Cl (250 mL), extracted with ethyl acetate, the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was washed with ethyl acetate/diethyl ether/n-hexane The alkane mixture was recrystallized to obtain intermediate 40-a (9.5 g, 30.95 mmol, 67.7% yield) as an off-white solid.

Step 2. Synthesis of Intermediate 40-b

To a mixture of 40-a (1 g, 2.21 mmol) and 36-a (830.31 mg, 2.43 mmol) in dioxane (5.00 mL)/H ₂ O (5.00 mL) was added Na ₂ CO ₃ (327.30 mg, 3.09 mmol), S-phos (108.51 mg, 264.67 μmol) and Pd(OAc) ₂ (35.35 mg, 154.39 μmol), the mixture was replaced with nitrogen several times, and then heated to 60 ° C under nitrogen protection, and at this temperature The reaction was stirred for 15 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated. The obtained crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and spin-dried to obtain the crude product using a silica gel column. Isolation and purification (PE/EA=1/1) gave intermediate 40-b (200 mg, 339.69 μmol, 15.40% yield) as a white solid. MS (ESI) m/z = 589 [M+H] ⁺ .

Synthesis of step 3 and step 4, intermediate Z-40

Referring to the method of step 2 and step 3 in the synthesis method of intermediate Z-39, using 40-b as raw material, through Pd/C reduction and DMP oxidation, intermediate Z-40 can be obtained, MS(ESI) m/z=497[ M+H] ⁺ .

Synthesis of Intermediate Z-41

Step 1. Synthesis of Intermediate 41-a

To a mixture of intermediates 40-a (469.02 mg, 1.46 mmol) and 37-a (600 mg, 1.32 mmol) in DMF (8 mL)/H ₂ O (2 mL) was added K ₃ PO ₄ (393.32 mg, 1.85 mmol) , S-phos (65.19mg, 159.01μmol) and Pd(OAc) ₂ (21.21mg, 92.63μmol), the mixture was replaced with nitrogen several times, heated to 100°C under the protection of nitrogen, and stirred for 15h at this temperature . After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, the obtained crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and spin-dried to obtain the crude product with silica gel column separation and purification (PE /EA=2/1) to obtain intermediate 41-a (130 mg, 228.58 μmol, 17.27% yield) as a white solid. MS (ESI) m/z = 383 [M+H] ⁺ .

Step 2. Synthesis of Intermediate Z-41

At room temperature, TFA (18.04 mg, 158.25 μmol) was added to a solution of 41-a (90 mg, 158.25 μmol) in DCM (2 mL), and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated and the crude product was redissolved. In DCM (5 mL), TEA (16.01 mg, 158.25 μmol, 22.07 μL) was added at room temperature, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated again and separated and purified by pre-HPLC to obtain intermediate Z-41 (50 mg, 130.76 μmol, 82.63% yield) as a white solid.

Synthesis of Intermediate Z-42

Step 1. Synthesis of Intermediate 42-a

To a solution of 3-(4-hydroxy-1-oxo-1,3-dihydroisoindol-2-yl)piperidine-2,6-dione (500 mg, 1.92 mmol) in DMF (8 mL) was added 4-Bromobutyloxy-tert-butyldimethylsilane (514mg, 1.92mmol), KHCO ₃ (570mg, 5.76mmol) and a catalytic amount of KI, the mixture was heated to 70°C, and the reaction was stirred at this temperature for 15h . After the completion of the reaction, the reaction solution was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered and spin-dried to obtain the crude product, which was separated and purified by mHPLC to obtain intermediate 42-a (160 mg, 0.48 mmol, 25% yield). MS (ESI) m/z = 333 [M+H] ⁺ .

Step 2. Synthesis of Intermediate Z-42

To a solution of 42-a (80 mg, 240.71 μmol) in DCM (10 mL) was added DMP (204.12 mg, 481.42 μmol), and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and the crude product was separated and purified by silica gel column (DCM/MeOH=10/1) to obtain intermediate Z-42 (75 mg, 227.04 μmol, 94.32% yield). MS (ESI) m/z=331 [M+H] ⁺ .

Synthesis of Intermediate Z-43

Step 1. Synthesis of Intermediate 43-b

43-a (5.0 g, 50.94 mmol) was dissolved in DMF (100 mL) solution, the temperature was lowered to 0 °C under nitrogen protection, NaH (2.04 g, 50.94 mmol) was added, the reaction was incubated for 30 minutes, and 3-benzyloxy bromide was added. Propane (9.34 g, 40.76 mmol), the reaction mixture was heated to 25°C and stirred for 2 h. After the reaction was completed, saturated aqueous ammonium chloride solution was added to quench, diluted with water, and extracted with EA. The combined organic phases were successively water and saturated brine. Washed, dried over anhydrous sodium sulfate, spin-dried, separated and purified by silica gel column (PE/EA=20/1) to obtain 43-b (2.5 g, 10.15 mmol, 19.92% yield). MS(ESI) m/z=247[M+1] ⁺ .

Step 2. Synthesis of Intermediate 43-c

1-b (20.0 g, 61.89 mmol) was dissolved in DMF (500 mL) solution, the temperature was lowered to 0 °C under nitrogen protection, NaH (2.72 g, 68.08 mmol) was added, the reaction was incubated for 30 minutes, and SEMCl (12.66 g, 75.83 mmol) was added. mmol), the reaction mixture was heated to 25 °C and stirred for 12 h. After the reaction was completed, saturated aqueous ammonium chloride solution was added to quench, diluted with water, extracted with EA, the combined organic phases were washed with water and saturated brine in turn, and anhydrous sodium sulfate After drying and spin drying, crude product 43-c (28 g, 61.76 mmol, 99.78% yield) was obtained. MS (ESI) m/z = 453 [M+1] ⁺ , used directly in the next step.

Step 3. Synthesis of Intermediate 43-d

To a solution of 43-c (6.91 g, 15.24 mmol) in DMF (100 mL) was added 43-b (4.13 g, 16.76 mmol), CuI (435.38 mg, 2.29 mmol), Pd(Ph ₃ P) ₂ Cl ₂ ( 801.24 mg, 1.14 mmol) and TEA (4.63 g, 45.72 mmol, 6.37 mL). The reaction solution was stirred at 85°C for 12 hours under the protection of N ₂ , diluted with water, extracted with EA, and the separated organic phase was washed with water and brine, dried and then spin-dried. The crude product was separated and purified by silica gel column (PE/EA=1/1) to obtain intermediate 43-d (6.66 g, 10.76 mmol, 70.62% yield). MS (ESI) m/z=619 [M+H] ⁺ .

Step 4. Synthesis of Intermediate 43-e

Pd/C (0.1 g, 10% purity) was added to a solution of 43-d (1000 mg, 1.62 mmol) in MeOH (20 mL), the reaction solution was replaced with hydrogen, and stirred at room temperature for 12 h under the protection of hydrogen. After the reaction was completed, filter , the filtrate was concentrated and dried to obtain the crude product of intermediate 43-e (600 mg, 1.13 mmol, 69.70% yield), which was directly used in the next reaction. MS (ESI) m/z = 533 [M+H] ⁺ .

Step 5. Synthesis of Intermediate Z-43

To a solution of 43-e (200 mg, 375.42 μmol) in DCM (10 mL) was added DMP (238.76 mg, 563.12 μmol), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was filtered, the filtrate was concentrated, and the crude product was separated and purified by silica gel column (DCM/MeOH=10/1) to obtain intermediate Z-43 (199 mg, 374.96 μmol, 99.88% yield). MS (ESI) m/z = 531 [M+H] ⁺ .

Synthesis of Intermediate Z-44

Referring to the synthetic route of intermediate Z-43, the method from step 1 to step 5, in step 1, 4-pentynyl-1-ol was used instead of 5-hexynyl-1-ol aniline as the starting material, The remaining operations are the same and go through the same steps to obtain the intermediate Z-44, MS(ESI) m/z=517[M+1] ⁺ .

Synthesis of Intermediate Z-45

Step 1. Synthesis of Intermediate 45-b

To a solution of 28-a (50 mg, 78.17 μmol) in DMF (5 mL)/H ₂ O (0.5 mL) was sequentially added 45-a (24.17 mg, 78.17 mmol), K ₂ CO ₃ (32.36 mg, 234.5 μmol), Pd(dppf)Cl2 (5.71 _mg , 7.82 μmol). The reaction solution was stirred at 90°C for 12 hours under the protection of N ₂ , diluted with water, extracted with ethyl acetate, and the separated organic phase was washed with water and brine, dried and then spin-dried. The crude product was isolated and purified by pre-TLC (100% EA) to give intermediate 45-b (57 mg, 76.82 mmol, 98.28% yield). MS(ESI) m/z=742[M+H] ⁺ .

Step 2. Synthesis of Intermediate Z-45

At room temperature, TFA (18.04 mg, 158.25 μmol) was added to a solution of 45-b (32 mg, 43.13 μmol) in DCM (2 mL), and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain the intermediate crude product. Body Z-45 (22 mg, 43.00 μmol, 99.71% yield) was used directly in the next step.

Synthesis of Intermediate Z-46

Step 1. Synthesis of Intermediate 46-a

Pd/C (3 mg, 10% purity) was added to a solution of 45-b (32 mg, 43.13 μmol) in MeOH (5 mL), the reaction solution was replaced with hydrogen several times, and then stirred at room temperature for 12 h under the protection of hydrogen. After the reaction was completed, After filtration, the filtrate was concentrated and dried to obtain the crude product of intermediate 46-a (32 mg, 43.01 μmol, 99.73% yield), which was directly used in the next reaction without purification. MS (ESI) m/z = 744 [M+H] ⁺ .

Step 2. Synthesis of Intermediate Z-46

At room temperature, TFA (18.04 mg, 158.25 μmol) was added to a solution of 46-a (33 mg, 44.35 μmol) in DCM (2 mL), and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated to obtain the intermediate crude product. Body Z-46 (22 mg, 42.83 μmol, 96.57% yield) was used directly in the next step.

Synthesis of Intermediate Z-47

Step 1. Synthesis of Intermediate Z-47

To a solution of 47-a (200 mg, 431.35 μmol) in DCM (10 mL) was added DMP (267.98 mg, 632.03 μmol), and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and the crude product was separated and purified by silica gel column (PE/EA=1/2) to obtain intermediate Z-47 (198 mg, 418.92 μmol, 99.42% yield). MS (ESI) m/z = 473 [M+H] ⁺ .

Synthesis of Intermediate Z-48

Step 1. Synthesis of Intermediate 48-c

To the NMP (10 mL) solution of 48-a (200 mg, 724.06 μmol) was added 48-b (100.07 mg, 838.87 μmol), DIPEA (280.73 mg, 2.17 mmol, 378.35 μL), and the mixture was heated to 90 ° C under nitrogen protection The reaction was carried out for 12 hours. After the reaction was completed, water was added, extracted with EA, the organic phase was concentrated, and the crude product was separated and purified by silica gel column (PE/EA=1/4) to obtain intermediate 48-c (200 mg, 538.52 μmol, 74.38% yield). MS (ESI) m/z = 372 [M+H] ⁺ .

Step 2. Synthesis of Intermediate Z-48

To a solution of 48-c (200 mg, 538.52 μmol) in DCM (10 mL) was added DMP (342.50 mg, 807.78 μmol), and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and the crude product was separated and purified by silica gel column (PE/EA=1/4) to obtain intermediate Z-48 (134 mg, 362.78 μmol, 67.37% yield). MS (ESI) m/z = 370 [M+H] ⁺ .

Synthesis of Intermediate Z-49

Step 1. Synthesis of Intermediate 49-a

2-b (3.0 g, 9.28 mmol) was dissolved in DMF (50 mL) solution, the temperature was lowered to 0 °C under nitrogen protection, NaH (445.63 mg, 11.14 mmol) was added, the reaction was incubated for 30 minutes, SEMCl (1.86 g, 11.14 mmol) was added. mmol), the reaction mixture was heated to 25 °C and stirred for 12 h. After the reaction was completed, saturated aqueous ammonium chloride solution was added to quench, diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine successively, and anhydrous Dry over sodium sulfate, spin dry, and separate and purify the crude product with silica gel column (PE/EA=1/1) to obtain intermediate 49-a (2.97 g, 6.55 mmol, 70.56% yield). MS(ESI) m/z=453[M+1] ⁺ .

Step 2. Synthesis of Intermediate 49-c

49-a (0.2 g, 441.11 μmol), 49-b (127.97 mg, 529.33 μmol), Pd(OAc) ₂ (3.96 mg, 17.64 μmol), BINAP (21.97 mg, 35.29 μmol), Cs ₂ CO ₃ ( 286.72 mg, 882.22 μmol) was dissolved in a mixed solvent of toluene (10 mL) and t-BuOH (1 mL), replaced with nitrogen, and heated to 100° C. to react for 12 hours. After the completion of the reaction, water was added, extracted with EA, the organic phase was washed with water and saturated brine successively, dried over anhydrous sodium sulfate, spin-dried, and the crude product was separated and purified with a silica gel column (100% EA) to obtain intermediate 49-c (40 mg, 69.23 μmol, 15.69% yield). MS(ESI) m/z=578[M+1] ⁺ .

Step 3. Synthesis of Intermediate 49-d

Pd/C (5 mg, 10% purity) was added to a solution of 49-c (45 mg, 77.88 μmol) in MeOH (5 mL), the reaction solution was replaced with hydrogen several times, and then stirred at room temperature for 4 h under the protection of hydrogen. After the reaction was completed, After filtration, the filtrate was concentrated and dried to obtain the crude product of intermediate 49-d (36 mg, 73.82 μmol, 94.87% yield), which was directly used in the next reaction without purification. MS (ESI) m/z=488 [M+H] ⁺ .

Step 4. Synthesis of Intermediate Z-49

To a solution of 49-d (36 mg, 73.82 μmol) in DCM (3 mL) was added DMP (46.95 mg, 110.73 μmol), and the mixture was stirred for 1 hour at room temperature. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and the crude product was separated and purified by silica gel column (100% EA) to obtain intermediate Z-49 (31 mg, 63.83 μmol, 86.47% yield). MS (ESI) m/z = 486 [M+H] ⁺ .

Synthesis of Intermediate Z-50

Step 1, Synthesis of 50-a

To a solution of 27-g (3.0 g, 25.64 mmol) in THF (100 mL)/H ₂ O (50 mL) at zero degrees, potassium carbonate (10.62 g, 76.92 mmol) and Cbz-OSu (12.82 g, 51.28 mmol) were sequentially added ), and stirred at room temperature for 1 hour. After the reaction was completed, saturated aqueous sodium bicarbonate solution was added, and EA was added to extract and separate the layers. The separated organic phase was washed with water and saturated brine successively, dried over anhydrous sodium sulfate and concentrated. The residue was washed with silica gel column. Isolation and purification (50% EA in PE) gave 50-a (5.1 g, 20.29 mmol, 79.13% yield). MS(ESI)m/z=252[M+1] ⁺

Step 2. Synthesis of 50-b

At zero temperature, to a solution of 50-a (5.1 g, 20.29 mmol) in DCM (100 mL), TEA (6.16 g, 60.88 mmol, 8.49 mL) and MsCl (3.49 g, 30.44 mmol) were sequentially added, and stirred at zero temperature for 1 hour After the reaction was completed, saturated aqueous sodium bicarbonate solution was added to separate the layers, and the separated organic phase was washed with water and saturated brine successively, dried over anhydrous sodium sulfate, and concentrated to obtain 50-b (6.66 g, 20.22 mmol, 99.63 % yield), the crude product was directly used in the next reaction without purification. MS(ESI)m/z=330[M+1] ⁺

Step 3, Synthesis of 50-d

50-b (6.68 g, 20.28 mmol), 50-c (5.52 g, 22.31 mmol), NaI ( _303.96 mg, 2.03 mmol) and K2CO3 (8.40 _g , 60.84 mmol) were dissolved in DMF (100 mL) , the reaction was raised to 80°C and the reaction was stirred overnight. After the reaction was completed, it was cooled to room temperature, EA was added, the mixture was washed twice with water, the separated organic phase was concentrated, and the residue was separated and purified with a silica gel column (50% EA in PE) to obtain 50-d (3.3 g, 6.87 mmol, 33.86% yield). MS(ESI)m/z=481[M+1] ⁺

Step 4. Synthesis of Intermediate 50-e

Pd/C (1.0 g, 10% purity) was added to the solution of 50-d (3.3 g, 6.87 mmol) in MeOH (60 mL), the reaction solution was replaced with hydrogen several times, and then stirred at room temperature for 4 h under the protection of hydrogen, the reaction was completed After that, it was filtered, and the filtrate was concentrated and dried to obtain the crude product of intermediate 50-e (2.16 g, 6.24 mmol, 90.80% yield), which was directly used in the next reaction without purification. MS (ESI) m/z = 347 [M+H] ⁺ .

Step 5, Synthesis of 50-g

In an ice bath, to a solution of 50-e (2.16 g, 6.24 mmol) in DCM (50 mL), DIPEA (2.42 g, 18.71 mmol) and 50-f (1.20 g, 6.24 mmol) were sequentially added, and the reaction was stirred overnight at room temperature. After completion, the reaction solution was concentrated, and the residue was separated and purified by normal phase silica gel column (PE/EA=1/1) to obtain 50-g (3.0 g, 5.96 mmol, 95.66% yield). MS(ESI)m/z=503[M+1] ⁺

Step 6, Synthesis of 50-h

To 50-g (3.82 g, 7.60 mmol) of EtOH (60 mL)/H ₂ O (10 mL), NH ₄ Cl (4.03 g, 75.95 mmol) and Fe (2.55 g, 45.57 mmol) were sequentially added, and stirred at 80 °C After 12 hours, the reaction was completed, filtered, the filtrate was rinsed with EA, and concentrated to obtain 50-h (3.15 g, 6.66 mmol, 87.69% yield). The crude product was used for the next reaction without purification. MS(ESI)m/z=473[M+1] ⁺

Step 7, Synthesis of 50-i

To a mixed solution of 50-h (100 mg, 211.43 μmol) in EtOH (3 mL), CNBr (111.06 mg, 1.06 mmol) was added, and the mixture was stirred at 60° C. overnight. After the reaction was completed, a saturated aqueous sodium bicarbonate solution was added, stirred for 10 minutes, and the layers were separated. The organic phase was washed with a saturated aqueous sodium bicarbonate solution, water, and brine in this order, dried over anhydrous sodium sulfate, and the organic phase was concentrated. The obtained residue was washed with a silica gel column. Isolation and purification (DCM/MeOH=10/1) gave 50-i (90 mg, 180.73 μmol, 85.48% yield). MS(ESI)m/z=498[M+1] ⁺

Step 8, Synthesis of 50-j

To 50-i (2.0 g, 4.02 mmol) in THF (50 mL)/H ₂ O (50 mL), NaOH (642.60 mg, 16.07 mmol) was added, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated and diluted with water. , adjusted to pH=4-5 with 1M HCl, a solid was precipitated, filtered, and the filter cake was washed with water, and dried to obtain 50-j (1.2 g, 2.48 mmol, 61.74% yield). MS(ESI) m/z=484[M+1] ⁺ .

Step 9, Synthesis of 50-k

To 50-j (1.23 g, 2.54 mmol) in DCM (5 mL), TEA (1.03 g, 10.17 mmol, 1.42 mL) and TBTU (979.02 mg, 3.05 mmol) were sequentially added, and stirred at room temperature for 15 minutes. After the reaction was completed, Add water to dilute, separate the layers, and concentrate the organic phase. The obtained residue is separated and purified by silica gel column (DCM/MeOH=10/1) to obtain the intermediate 50-k (1.0 g, 2.15 mmol, 84.44% yield). MS(ESI) m/z=466[M+1] ⁺ .

Step 10, the synthesis of intermediate 50-1

To a solution of 50-k (1.0 g, 2.15 mmol) in DMF (30 mL), DIPEA (832.12 mg, 6.44 mmol, 1.12 mL) and SEMCl (358.42 mg, 2.15 mmol) were sequentially added, and the reaction mixture was warmed to 60 °C and stirred 12h, after the reaction was completed, the reaction solution was concentrated, the residue was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine successively, dried over anhydrous sodium sulfate, and then spin-dried and separated and purified with a normal-phase silica gel column ( 100% ethyl acetate) to give 50-l (630 mg, 1.06 mmol, 49.24% yield). MS(ESI) m/z=596[M+1] ⁺ .

Step 11. Synthesis of Intermediate 50-m

To a solution of 50-l (300 mg, 503.19 μmol) in toluene (4 mL)/tert-butanol (1 mL) was added 1-tert-butoxycarbonylpiperazine (140.58 mg, 754.79 μmol), BINAP (25.07 mg, 40.26 μmol), Pd(OAc) ₂ (4.51 mg, 20.13 μmol) and Cs ₂ CO ₃ (327.07 mg, 1.01 mmol). Under nitrogen, the temperature was raised to 120°C and the reaction was stirred at this temperature for 15 hours. After the reaction was completed, the mixture was diluted with ethyl acetate, washed with water, and the organic phase was concentrated to obtain a crude product. The crude product was separated and purified by a reverse-phase silica gel column to obtain 50-m (370 mg, 495.99 μmol, 98.57% yield). MS (ESI) m /z=746[M+1] ⁺ .

Step 12. Synthesis of Intermediate Z-50

To a solution of 50-m (390 mg, 522.80 μmol) in DCM (10 mL) was added TFA (10 mL), and the mixture was stirred at room temperature for 1 hour. Then, the reaction solution was concentrated, and the residual TFA was removed with an oil pump to obtain Z-50 (269 mg, 521.71 μmol, 99.79% yield), which was directly used in the next reaction without purification. MS(ESI) m/z=516[M+1] ⁺ .

Synthesis of Intermediate Z-51

Step 1. Synthesis of Intermediate 51-a

At zero temperature, NaH (1.43 g, 59.42 mmol) was added to a solution of 1-b (16 g, 49.51 mmol) in DMF (470 mL), and the mixture was stirred for 1 hour at zero temperature, and then SEMCl (9.08 g, 54.47 mmol) was added. After dripping, the mixture was reacted at room temperature for 12h. After the reaction was completed, it was diluted with water, extracted with EtOAc, the combined organic phases were washed with water and brine, dried over anhydrous sodium sulfate, and concentrated to obtain crude 51-a (22 g, 48.52 mmol, 98.00% yield). MS(ESI) m/z=425[M-27] ⁺ .

Step 2 to Step 4, Synthesis of Intermediate Z-51

Referring to the operations of steps 3, 4 and 5 of the synthetic route of intermediate Z-1, intermediate 1-b is replaced by intermediate 51-a, and the remaining operations are the same to obtain intermediate Z-51. MS(ESI) m/z=445[M+1] ⁺ .

Synthesis of Intermediate Z-52

Referring to the synthesis method of intermediate Z-42, in step 1, 4-bromobutyloxy-tert-butyldimethylsilane was replaced with 4-bromobutyloxy-tert-butyldimethylsilane, and the rest of the operations were the same , the intermediate Z-52 can be obtained. MS(ESI) m/z=345[M+1] ⁺ .

Synthesis of Intermediate Z-53

Referring to the methods of steps 11 and 12 in the synthetic route of intermediate Z-50, in step 11, 1-Boc-4-(piperidin-4-yl)-piperazine was used instead of 1-tert-butoxycarbonylpiperazine, and the remaining operations The same, intermediate Z-53 can be obtained, MS(ESI) m/z=599[M+1] ⁺ .

Synthesis of Intermediate Z-54

Referring to the method of steps 11 and 12 in the synthetic route of intermediate Z-50, in step 11, intermediate 28-a is used instead of intermediate 50-1, and 4-piperazin-1-ylpiperidine-1-carboxylic acid is used at the same time The tert-butyl ester was replaced with 1-tert-butoxycarbonylpiperazine, and other operations were the same to obtain the intermediate Z-54, MS(ESI) m/z=598[M+1] ⁺ .

Synthesis of Intermediate Z-55

Referring to the procedure of steps 5 to 12 in the route of intermediate Z-50, in step 5, replace 2,6-dichloro-3-nitropyridine with 2,4-dichloro-5-nitropyridine, in step 11 Substitute 4-piperazin-1-ylpiperidine-1-carboxylic acid tert-butyl ester for 1-tert-butoxycarbonylpiperazine, and the other operations are the same to obtain intermediate Z-55. MS(ESI) m/z=598[M+1] ⁺ .

Synthesis of Intermediate Z-56

Referring to the method of step 11 and step 12 in the synthetic route of intermediate Z-50, in step 11, replace 1-tert-butoxycarbonylpiperazine with tert-butyl 4-piperazin-1-ylpiperidine-1-carboxylate , and other operations are the same to obtain the intermediate Z-56. MS(ESI) m/z=599[M+1] ⁺ .

Synthesis of Intermediate Z-57

Referring to the operation of step 11 and step 12 in the synthetic route of intermediate Z-50, intermediate 50-1 was replaced with intermediate 28-a, and 4-[(piperazin-1-yl)methyl]piperidine- Substitute 1-tert-butoxycarbonylpiperazine with 1-tert-butyl carboxylate, and the other operations are the same to obtain intermediate Z-57. MS(ESI) m/z=612[M+1]+.

Synthesis of Intermediate Z-58

Referring to the operations of step 7 and step 8 in the synthetic route of intermediate Z-55, replace 4-piperazin-1-ylpiperidine-1-carboxylate tert-butyl ester with 1-tert-butoxycarbonylpiperazine, and the remaining operations are the same , the intermediate Z-58 can be obtained. MS(ESI) m/z=516[M+1] ⁺ .

Synthesis of Intermediate Z-59

Step 1. Synthesis of Intermediate 59-a

Referring to the synthesis method of intermediate Z-25, using 4-c instead of intermediate 1-c, and other operating conditions are the same, intermediate 59-a can be obtained.

Step 2. Synthesis of Intermediate Z-59

Referring to the synthesis method of the intermediate Z-26, using 59-a instead of Z-25 as the starting material, the same operation and conditions, the intermediate Z-59 can be obtained.

Synthesis of Intermediate Z-60

Referring to the method from step 1 to step 4 of the synthetic route of intermediate Z-43, in step 1, 43-a was replaced with 7 octynyl alcohol as the starting material, and the rest of the operation steps were the same to obtain intermediate Z-60, MS (ESI )m/z=559[M+1] ⁺ .

Synthesis of Intermediate Z-61

Referring to the method from step 2 to step 4 of the synthetic route of Example Z-1, in the initial step, 10-undenyl alcohol was used to replace 4-pentyn-1-ol, and the rest of the operation steps were the same to obtain intermediate Z-61 . MS(ESI) m/z=543[M+1] ⁺ .

Synthesis of Intermediate Z-62

Referring to steps 11 and 12 in the synthetic route of intermediate Z-50, in the initial step, 3-(piperazin-1-ylmethyl)pyrrolidine-1-carboxylate tert-butyl ester was used to replace 1-tert-butoxycarbonyl group Piperazine, the remaining operation steps are the same, and intermediate Z-62 can be obtained. MS(ESI) m/z=599[M+1] ⁺ .

Synthesis of Intermediate Z-63

step 1

Z-63-1a (2g, 9.2mmol) and pyrazole (755mg, 11mmol) were mixed in acetonitrile (20ml), cooled in an ice-water bath (0℃-5℃), and acetyl chloride (872mg, 11mmol) was added dropwise, and then reacted at room temperature 17h, Z-63-1b (1.5g, 9.2mmol), pyrazole (1.4g, 19.3mmol) and triphenyl phosphite (3.4g, 11mmol) were directly added to the reaction solution the next day, and then heated at 85°C to react 17h, after the reaction was completed, cooled to room temperature, water was added to the reaction solution, a solid was precipitated, the solid was filtered and washed once with water and once with ethyl acetate, then the solid was filtered, spin-dried, and used directly in the next step.

Step 2

Z-63-1c (1.58g, 4.5mmol) was dissolved in DMF, cooled in an ice-water bath (0°C-5°C), sodium hydride (130mg, 5.4mmol) was added, the reaction was maintained at this temperature for 1h, and then SEMCl (906mg) was slowly added dropwise , 5.4 mmol), warmed to room temperature and reacted for 3 h. After the reaction, water was added, extracted three times with ethyl acetate, the organic layer was washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and used directly in the next step.

Step 3

Z-63-1d (500 mg, 1.04 mmol) and 4-pentyn-1-ol (105 mg, 1.25 mmol) were dissolved in DMF, then Pd(pph ₃ ) ₂ Cl ₂ (55 mg, 0.078 mmol), CuI ( 30mg, 0.156mmol) and TEA (315mg, 3.12mmol), heated at 80°C for 17 hours, after the reaction was completed, water was added, EA was added to extract the liquid, the separated organic phase was washed with water in turn, washed with saturated brine, and anhydrous sulfuric acid. After drying over sodium, it was concentrated, and the crude product was separated and purified by silica gel column (50% EA in PE) to give Z-63-1e (240 mg, 0.496 mmol, 47.8% yield). MS(ESI)m/z=484[M+1] ⁺

Step 4

Z-63-1e (240 mg, 0.496 mmol) was dissolved in methanol, 10% palladium on carbon (35 mg) was added, and the reaction was carried out at room temperature for 17 h under the protection of hydrogen. -1f (220 mg, 0.451 mmol, 90.1% yield) MS (ESI) m/z=488[M+1] ⁺

Step 5

Z-63-1f (220mg, 0.451mmol) was dissolved in dichloromethane, cooled in an ice bath, DMP (229mg, 0.54mmol) was added, then warmed to room temperature and reacted for 4h. After the reaction was completed, the solid was filtered out, the filtrate was spin-dried, and the crude product After column purification (PE:EA=2:1), compound Z-63 (120 mg, 0.246 mmol, 52% yield) was obtained, MS (ESI) m/z=486[M+1] ⁺

Synthesis of Intermediate Z-64

To a solution of 2-(2,6-dioxo-piperidin-3-yl)-4-hydroxy-isoindole-1,3-dione (1.57 g, 5.7 mmol) in DMF (50 mL) was added carbonic acid Potassium (1.19 g, 8.58 mmol) and tert-butyl bromoacetate (1.1 g, 5.71 mmol). After reacting at room temperature for 2 h, DMF was removed by rotary evaporation, ethyl acetate was dissolved, washed with water, and the organic phase was rotary dried. The crude product was purified by silica gel column (petroleum ether/ethyl acetate=2/1) to obtain the intermediate 2-(2,6-dioxo-piperidin-3-yl)-4-hydroxyacetic acid tert-butyl ester-isoindole -1,3-Dione (2 g, 5.1 mmol). It was dissolved in dichloromethane (15 mL), trifluoroacetic acid (10 mL) was added, reacted at room temperature for 1 h, and then the solvent was spin-dried to obtain the target compound Z-64 (1.6 g).

Synthesis of Intermediate Z-65

According to the synthesis method of Z-63, the starting material 2-amino-6-bromobenzoic acid was replaced with 2-amino-5-bromobenzoic acid, and other reagents and operation methods were the same to obtain intermediate Z-65.

Synthesis of Intermediate Z-66

Referring to the method from step 3 to step 5 in the synthetic route of intermediate Z-1, in the initial step, 3-butyn-1-ol was used to replace 4-pentyn-1-ol, and the rest of the operation steps were the same to obtain the intermediate Z-66. MS(ESI) m/z=315[M+1] ⁺ .

Synthesis of Intermediate Z-67

Referring to the method from step 1 to step 4 of the synthetic route of intermediate Z-36, in the initial step, 2-benzyloxybromoethane was used to replace 3-benzyloxybromopropane, and the rest of the operation steps were the same to obtain intermediate Z- 67. MS(ESI) m/z=353[M+1] ⁺ .

Synthesis of Intermediate Z-68

Referring to the synthesis method of intermediate Z-25, substituting 3-c for 1-c, and other reagents and operations are the same, intermediate Z-68 can be obtained. MS(ESI) m/z=341[M+1] ⁺ .

Example 1. Synthesis of compound 1

To Z-28 (30mg, 56.75μmol) in MeOH (2mL), add TEA to pH=9-10, then add Z-2 (18.63mg, 56.75μmol), stir at room temperature for 30 minutes, then add acetic acid to adjust pH= 4-5. After stirring for 5 minutes, NaBH ₃ CN (10.73 mg, 170.25 μmol) was added in one portion and the reaction mixture was overnight at room temperature. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated and purified by Pre.HPLC to obtain Example 1 (1.42 mg, 1.59 μmol, 2.80% yield, 94% purity). MS(ESI) m/z=837[M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.43(s, 1H), 7.96(s, 1H), 7.69-7.62(m, 1H ), 7.58(d, J＝1.2Hz, 1H), 7.53-7.45(m, 4H), 7.44(s, 1H), 7.39-7.31(m, 2H), 7.23-7.15(m, 1H), 5.12- 5.01(m,1H),4.49–4.25(m,3H),4.25–4.15(m,2H),4.09–3.85(m,3H),3.73(s,4H),3.56(s,2H),2.97– 2.82 (m, 2H), 2.82–2.72 (m, 1H), 2.73–2.59 (m, 4H), 2.56 (s, 3H), 2.49–2.29 (m, 5H), 2.30–2.16 (m, 3H), 2.08–1.89 (m, 4H), 1.69–1.55 (m, 2H), 1.44 (t, J=7.4Hz, 3H), 1.26 (dd, J=14.6, 7.1Hz, 2H), 0.80 (d, J= 6.5Hz, 3H).

Example 2, the synthesis of compound 2

Z-28 (46.60mg, 88.15μmol) and Z-25 (30mg, 88.15μmol) were mixed in DMF (5mL), then DIPEA (56.96mg, 440.75μmol, 76.77μL) and HATU (40.22mg, 105.78μL) were added sequentially μmol). The reaction solution was stirred at room temperature for 4 hours. After the reaction was completed, 1 mL of MeOH was added, and the crude product obtained by concentrating the reaction solution was separated and purified by Pre.HPLC to obtain Example 2 (1.32 mg, 1.37 μmol, 1.55% yield). MS(ESI) m/z=851[M+1] ⁺ , ¹ H NMR (400 MHz, CD ₃ OD) δ 8.86(s, 1H), 8.24(s, 1H), 8.01(s, 1H), 7.78 –7.68(m,2H),7.62–7.55(m,2H),7.49(d,J=7.6Hz,1H),7.38(d,J=8.2Hz,1H),5.18(dd,J=13.3,5.2 Hz, 1H), 4.57–4.29 (m, 6H), 4.13 (s, 1H), 3.96–3.76 (m, 6H), 2.99–2.71 (m, 10H), 2.50 (m, 1H), 2.33 (m, 1H), 2.24–2.12(m, 1H), 2.06(m, 2H), 1.57(m, 1H), 0.90(d, J=5.4Hz, 3H).

Example 3, the synthesis of compound 3

Referring to the synthesis method of Example 1, using intermediate compounds Z-28 and Z-16 as raw materials, through reductive amination, Example 3 was obtained. MS(ESI) m/z=851[M+1] ⁺ , ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.43(s, 1H), 7.95(s, 1H), 7.76-7.69(m, 1H ), 7.67–7.60 (m, 1H), 7.58 (s, 1H), 7.58–7.51 (m, 1H), 7.54–7.45 (m, 4H), 7.23–7.15 (m, 2H), 5.12 (dd, J = 13.2, 5.1Hz, 1H), 4.52–4.42 (m, 2H), 4.40–4.27 (m, 3H), 4.25–4.15 (m, 2H), 4.05–3.96 (m, 1H), 3.73 (s, 3H) ), 3.57(s, 4H), 2.82–2.67(m, 1H), 2.56(s, 4H), 2.47–2.28(m, 16H), 2.27–2.12(m, 1H), 2.10–1.87(m, 5H ), 1.67–1.52 (m, 9H), 1.53–1.40 (m, 2H), 1.32–1.18 (m, 7H), 0.90–0.81 (m, 1H), 0.80 (d, J=6.4Hz, 3H).

Example 4, the synthesis of compound 4

Referring to the synthesis method of Example 1, using intermediate Z-14 and intermediate Z-28 as raw materials, through reductive amination to obtain Example 4. MS(ESI) m/z=837[M+1] ⁺ , ¹ H NMR (400MHz, DMSO-d ₆ )δ8.43(s,1H),7.95(s,1H),7.63(s,1H), 7.57(d,J=1.2Hz,1H),7.55–7.45(m,4H),7.19(dd,J=8.3,1.3Hz,1H),5.08(dd,J=13.2,5.1Hz,1H),4.50 –4.40(m,2H),4.40–4.28(m,2H),4.25–4.15(m,1H),4.04–3.87(m,2H),3.73(s,3H),3.61(s,1H),3.57 (s, 3H), 2.97–2.82 (m, 1H), 2.82–2.72 (m, 2H), 2.71–2.57 (m, 2H), 2.56 (s, 3H), 2.52–2.43 (m, 4H), 2.44 –2.30(m,10H),2.26-2.15(m,2H),2.10-1.85(m,4H),1.79-1.65(m,3H),1.52-1.41(m,2H),0.81(d,J= 6.5Hz, 3H).

Example 5, the synthesis of compound 5

Referring to the synthesis method of Example 2, using intermediates Z-26 and Z-28 as raw materials and using HATU as a condensing agent, Example 5 is obtained through condensation reaction. MS(ESI) m/z=855[M+1] ⁺ , ¹ H NMR(400MHz, DMSO)δ12.71(s,1H),11.00(s,1H),8.43(s,1H),7.92(s ,1H),7.64–7.31(m,5H),7.18(d,J＝8.4Hz,1H),5.14(dd,J＝13.3,5.1Hz,1H),4.46(d,J＝17.1Hz,1H) ,4.33(dd,J＝23.3,11.0Hz,2H),4.20(d,J＝11.2Hz,1H),4.10–3.87(m,6H),3.73(s,4H),3.57(s,2H), 3.43(s, 5H), 3.00–2.85(m, 2H), 2.80(s, 2H), 2.71–2.61(m, 3H), 2.56(s, 3H), 2.38–2.25(m, 6H), 2.21( s, 1H), 2.09–1.87 (m, 3H), 1.67–1.42 (m, 5H), 0.81 (d, J=6.4Hz, 2H).

Example 6, the synthesis of compound 6

Referring to the synthesis method of Example 1, using Z-1 and Z-28 as raw materials, through reductive amination to obtain Example 6, MS (ESI) m/z=841[M+1] ⁺ , ¹ H NMR (400MHz, DMSO)δ12.70(s,1H),10.99(s,1H),8.42(s,1H),7.92(s,1H),7.65–7.52(m,2H),7.51–7.35(m,4H), 7.16(d,J＝8.9Hz,1H),5.13(dd,J＝13.2,5.1Hz,1H),4.45(d,J＝17.2Hz,1H),4.33(dd,J＝25.6,10.8Hz,2H) ),4.19(d,J＝11.0Hz,1H),3.98(d,J＝16.6Hz,2H),3.73(s,3H),3.54(s,2H),3.01–2.85(m,1H),2.79 (s, 1H), 2.63 (m, 3H), 2.56 (s, 3H), 2.44–2.27 (m, 7H), 2.28–2.12 (m, 4H), 2.06–1.85 (m, 3H), 1.67–1.53 (m, 2H), 1.44 (m, 3H), 1.37–1.25 (m, 2H), 0.80 (d, J=6.5Hz, 3H).

Synthesis of Examples 7-27 and Compounds 7-27

Referring to the reductive amination method of Example 1, Z-2 in the step was replaced with the corresponding SM-aldehyde in the table. Substitute Z-28 in the step with SM-amine, and the remaining methods are the same to obtain the following examples 7 to 27:

Example 27, the synthesis of compound 27

Step 1. Synthesis of Intermediate 27-1

To a solution of intermediate Z-35 (0.9 g, 2.66 mmol) in DMF (10 mL) was added 4-pentyn-1-ol (448 mg, 5.32 mmol), TEA (806 mg, 7.98 mmol), CuI (101 mg, 0.5 mmol) ) and Pd(PPh ₃ ) ₂ Cl ₂ (187 mg, 0.27 mmol), the mixture was replaced with nitrogen several times, then heated to 90° C. under the protection of N ₂ , and the reaction was stirred for 12 h at the temperature again. After the reaction was completed, the reaction solution was concentrated, the crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and the concentrated crude product was separated and purified by silica gel column (100% EA), 27-1 was obtained (190 mg, 0.56 mmol, 21% yield), MS (ESI) m/z = 342 [M+1] ⁺ .

Step 2. Synthesis of Intermediate 27-2

To a solution of 27-1 (190 mg, 0.56 mmol) in DCM (30 mL) was added DMP (476 mg, 1.12 mmol) and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and the obtained crude product was separated and purified by silica gel column (DCM/MeOH=10/1) to obtain 27-2 (134 mg, 0.39 mmol, 71% yield). MS (ESI) m/z=340 [M+H] ⁺ .

Step 3. Synthesis of compound 27

27-2 (58.4 mg, 0.17 mmol) and catalytic amount of acetic acid were added to intermediate Z-28 (70 mg, 0.13 mmol) in DCM (7 mL), the mixture was stirred at room temperature for 1 h, and then NaBH(OAc) ₃ (84 mg) was added. , 0.397mmol), and continued to stir the reaction at room temperature for 4h. After the reaction was completed, the reaction solution was concentrated, the crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and the concentrated crude product was separated and purified by mHPLC to obtain compound 27 (45 mg, 0.05 mmol). , the yield is 40%). MS (ESI) m/z = 852 [M+H] ⁺ .

Example 28, the synthesis of compound 28

Referring to the method of step 1 to step 3 in the synthesis of Example 27, in step 1, intermediate Z-33 is used instead of intermediate Z-35, and the other methods remain unchanged, and compound 28 can be obtained. MS (ESI) m/z = 852 [M+H] ⁺ .

Example 29, the synthesis of compound 29

Referring to the method of step 1 to step 3 in the synthesis of Example 27, in step 1, intermediate Z-34 is used instead of intermediate Z-35, and the other methods remain unchanged, and compound 29 can be obtained. MS (ESI) m/z = 852 [M+H] ⁺ .

Synthesis of Example 30 to Example 38 and Compound 30 to Compound 38

Refer to the method for reductive amination of step 3 in the synthesis of Example 27, and replace 27-2 in the step with the corresponding SM-aldehyde in the table. Substitute SM-amine for Z-30 in the step, and the remaining methods are the same to obtain the following examples 30 to 38:

Example 39, the synthesis of compound 39

Step 1. Synthesis of Intermediate 39-1

To a mixed solution of 27-1 (230 mg, 0.67 mmol) in MeOH (15 mL)/THF (5 mL) was added Pd/C (200 mg). After the mixture was replaced with hydrogen, the reaction was stirred at room temperature for 16 hours under the protection of hydrogen. After the reaction was completed, Pd/C was removed by filtration, and the filtrate was concentrated to give 39-1 (181 mg, 0.52 mmol, 78% yield, crude product).

Step 2. Synthesis of Intermediate 39-2

To a solution of compound 39-1 (181 mg, 0.52 mmol) in DCM (6 mL) was added DMP (331 mg, 0.78 mmol). The suspension was stirred at room temperature for 1.5 hours. After the reaction was completed, the mixture was filtered, the filtrate was concentrated and spin-dried, the crude product was diluted with ethyl acetate, a brown solid was precipitated, the solid was collected by filtration, and separated and purified with a reverse-phase C18 column to obtain an intermediate compound 39-2 (116 mg, 0.34 mmol, 65% yield). MS(ESI) m/z=344[M+1] ⁺ .

Step 3. Synthesis of compound 39

Referring to the method of step 3 of Example 27, the intermediate 39-2 was used instead of 27-2, and the remaining operation methods were the same (i.e. NaBH(AcO) ₃ was used as a reagent, DCM was added with catalytic amount of acetic acid as a solvent, and reductive amination was performed), and the obtained Compound 39. MS(ESI) m/z=856[M+1] ⁺ .

Example 40, the synthesis of compound 40

Referring to the method of step 1 to step 3 in the synthesis of Example 39, in step 1, intermediate 40-2 is used instead of intermediate 39-2, and the other methods remain unchanged, and compound 28 can be obtained. MS(ESI) m/z=856 [M+H] ⁺ .

Example 41, the synthesis of compound 41

Referring to the methods from step 1 to step 3 in the synthesis of Example 39, in step 1, intermediate 27-1 is replaced by intermediate 29-1, and the other methods remain unchanged, and compound 41 can be obtained. MS(ESI) m/z=856 [M+H] ⁺ .

Synthesis of Example 42 to Example 50 and Compound 42 to Compound 50

Refer to the method for reductive amination of step 3 in the synthesis of Example 27, and replace 27-2 in the step with the corresponding SM-aldehyde in the table. Substitute SM-amine for Z-30 in the step, and the remaining methods are the same to obtain the following examples 42 to 50:

Example 51, synthesis of compound 51 Z-28

Step 1. Synthesis of Intermediate 51-1

To a mixture of Z-28 (42.58 mg, 80.54 μmol) and Z-40 (42.58 mg, 80.54 μmol) in DCM (2 mL), TFA was added dropwise to adjust the pH to 5-6, and after stirring for 30 min, NaBH (OAc ) ₃ (51.21 mg, 241.63 μmol), the mixture was stirred and reacted at room temperature for 15 hours. After the completion of the reaction, the reaction solution was diluted with water, extracted with DCM, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and the concentrated crude product was separated and purified by pre-HPLC to obtain 51-1 (12 mg, 11.89 μmol, 14.76% yield) as a yellow solid.

Step 2. Synthesis of compound 51

To a solution of 51-1 (12 mg, 11.89 μmol) in DCM (2 mL) was added TFA (1.36 mg, 11.89 μmol, 2 mL) at room temperature, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated, the obtained crude product was redissolved in DCM (2 mL), TEA (725.50 mg, 7.17 mmol, 1 mL) was added at room temperature, and the reaction was stirred for 30 minutes. The mixture was concentrated, and the crude product was separated and purified by pre-HPLC to obtain 51 (1.14 mg, 1.14 umol, 9.56% yield, 99% purity, TF) as a yellow solid. MS (ESI) m/z = 879 [M+H] ⁺ . Nuclear magnetic spectrum: ¹ H NMR (400MHz, DMSO-d6)δ12.81(s,1H), 10.99(s,1H), 8.46(s,1H), 8.34(s,1H), 8.05(s,1H) ,7.97(s,1H),7.81(s,1H),7.72(d,J=3.1Hz,2H),7.57(d,J=30.4Hz,3H),7.25(s,1H),5.11(dd, J=13.3, 5.2Hz, 1H), 4.46 (d, J=17.2Hz, 1H), 4.34 (d, J=17.4Hz, 2H), 4.29–4.15 (m, 3H), 4.02 (s, 1H), 3.74(s, 4H), 3.43(s, 28H), 2.80(s, 3H), 2.58(s, 7H), 2.17(s, 4H), 1.96(d, J＝36.6Hz, 4H), 1.46(s ,1H),1.06(t,J＝7.0Hz,2H),0.82(d,J＝6.4Hz,3H).

Example 52, the synthesis of compound 52

Referring to the synthesis steps 1 and 2 of compound 51, using intermediate Z-30 instead of Z-28 in step 1 as the starting material, and the rest of the operation method is the same, compound 52 can be obtained. MS (ESI) m/z = 879 [M+H] ⁺ . Nuclear magnetic spectrum: ¹ H NMR (400MHz, DMSO-d ₆ )δ12.87(s, 1H), 8.48(s, 1H), 8.34(s, 1H), 8.02(d, J=20.8Hz, 2H), 7.86–7.50 (m, 6H), 7.33 (d, J=8.3Hz, 1H), 5.11 (dd, J=13.2, 5.1Hz, 1H), 4.47 (d, J=17.2Hz, 1H), 4.41–4.30 (m, 2H), 4.22 (q, J=5.7, 5.2Hz, 3H), 4.08–3.90 (m, 4H), 3.75 (s, 5H), 2.92 (ddd, J=18.2, 13.8, 5.6Hz, 4H ), 2.80(s, 2H), 2.60(s, 5H), 2.40(td, J=13.3, 4.7Hz, 2H), 2.19(d, J=28.0Hz, 3H), 2.06–1.90(m, 3H) ,1.46(s,1H),1.23(s,1H),0.82(d,J＝6.4Hz,3H).

Example 53, the synthesis of compound 53

Referring to the synthesis method of step 1 in Example 51, Z-28 and Z-36 were used as raw materials for reductive amination to obtain compound 53, MS (ESI) m/z=879 [M+H] ⁺ . Nuclear magnetic spectrum: ¹ H NMR (400MHz, DMSO-d ₆ )δ12.84(s,1H), 11.04(s,1H), 8.50(s,1H), 8.26(s,1H), 8.03(d,J =4.5Hz,2H),7.89(dd,J=7.6,1.2Hz,1H),7.67(s,1H),7.64-7.59(m,2H),7.59-7.52(m,2H),7.30-7.24( m, 1H), 5.26–5.15 (m, 1H), 4.65–4.54 (m, 3H), 4.46–4.32 (m, 5H), 4.30–4.18 (m, 5H), 4.07–3.97 (m, 3H), 3.94–3.82 (m, 2H), 3.75 (s, 4H), 3.05–2.89 (m, 4H), 2.86–2.73 (m, 2H), 2.70–2.62 (m, 1H), 2.62 (s, 3H), 2.29–2.09 (m, 5H), 2.09–1.84 (m, 4H), 1.51–1.38 (m, 1H), 0.82 (d, J=6.5Hz, 3H).

Synthesis of Example 54 to Example 62, Compound 54 to Compound 62

Referring to the method of step 1 of Example 51, the SM-aldehyde and SM-amine in the following table are used as raw materials, and the corresponding example compounds in the table can be obtained by reductive amination.

Example 63, the synthesis of compound 63

To a solution of Z-28 (30 mg, 56.75 μmol) in DMF (2 mL) was added Z-37 (21.70 mg, 56.75 μmol), DIPEA (36.67 mg, 283.74 μmol, 49.42 μL) and HATU (21.56 mg, 56.75 μmol), The reaction mixture was stirred at room temperature overnight.

After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated and separated and purified by pre-HPLC to obtain compound 63 (9.38 mg, 0.009 mmol, 16% yield, 99.4% purity, TF) as a yellow solid, MS (ESI) m/z=893 [M+H] ⁺ .

1H NMR(400MHz,Methanol-d ₄ )δ8.89(s,1H),8.27(s,1H),8.06(d,J=13.1Hz,2H),7.78(t,J=6.6Hz,2H), 7.70–7.59 (m, 3H), 7.51 (t, J=7.3Hz, 1H), 7.39 (d, J=6.9Hz, 1H), 5.25–5.16 (m, 1H), 4.67–4.39 (m, 8H) ,4.31(d,J＝13.8Hz,1H),3.90(d,J＝11.8Hz,1H),3.85(d,J＝1.3Hz,4H),3.05(s,4H),2.83(s,4H) , 2.77(s, 2H), 2.58(tt, J＝13.3, 6.7Hz, 2H), 2.32(s, 1H), 2.23–2.15(m, 1H), 2.03(d, J＝12.2Hz, 2H), 1.55(d, J=10.3Hz, 1H), 0.85(t, J=6.5Hz, 3H).

Synthesis of Example 64 to Example 74, Compound 64 to Compound 74

Referring to the method of Example 63, SM-amine and SM-acid in the following table are used as raw materials, and the corresponding example compounds in the table can be obtained through condensation reaction under the same conditions.

Example 75, the synthesis of compound 75

Z-42 (31.24 mg, 94.58 μmol) was added to a solution of Z-30 (50 mg, 94.58 μmol) in DCM (4 mL), the reaction mixture was stirred at room temperature for 30 min, and then AcOH was added dropwise to adjust the pH of the reaction solution to 3 to 4. Then NaBH(AcO) ₃ (60.14 mg, 283.74 μmol) was added, and the mixture was stirred at room temperature for 5 hours. After the reaction was completed, the reaction solution was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and the crude product obtained after concentration was separated and purified by Pre.HPLC to obtain compound 75 (33.8 mg , 35.32 μmol, 37.34% yield, TF), MS (ESI) m/z=843 [M+H]+. Nuclear magnetic spectrum: ¹ H NMR (400MHz, DMSO) δ12.86(s, 1H), 10.99(s, 1H), 8.50(s, 1H), 8.01(s, 1H), 7.65(d, J=8.2Hz ,2H),7.51(dd,J＝19.0,11.2Hz,2H),7.33(d,J＝7.5Hz,2H),7.25(d,J＝8.2Hz,1H),5.13(dd,J＝13.3, 5.0Hz, 1H), 4.37 (dd, J=11.1, 6.1Hz, 5H), 4.30–4.11 (m, 10H), 3.98 (dd, J=27.2, 16.7Hz, 7H), 3.75 (s, 4H), 3.01–2.86 (m, 3H), 2.80 (s, 2H), 2.71–2.55 (m, 5H), 2.23 (s, 2H), 2.06–1.88 (m, 4H), 1.79 (s, 5H), 1.46 ( s,1H),0.82(d,J＝6.4Hz,3H).

Synthesis of Example 76 to Example 78, Compound 76 to Compound

Referring to the method of Example 75, SM-amine and Z-42 in the following table are used as raw materials, and the corresponding example compounds in the table can be obtained through reductive amination reaction under the same conditions.

Example 79 to Example 81, Synthesis of Compound 79 to Compound 81

Referring to the method of Example 75, using the intermediate Z-28 as the raw material and the aldehyde of the corresponding structure in the following table as the raw material, the corresponding example compounds in the table can be obtained through reductive amination reaction under the same conditions.

Example 82, Synthesis of Compound 82

Referring to the method of Example 63, using the intermediate Z-28 and the SM-acid in the following table as raw materials, the compound 82 of Example can be obtained by condensation reaction under the same conditions.

Example 83 to Example 85, Synthesis of Compound 83 to Compound 85

Referring to the method of Example 75, using the intermediate Z-30 as the raw material and the aldehyde of the corresponding structure in the following table as the raw material, the corresponding example compounds in the table can be obtained through reductive amination reaction under the same conditions.

Example 86 to Example 91, Synthesis of Compound 86 to Compound 91

Referring to the method of Example 75, using the intermediate Z-31 as the raw material and the aldehyde of the corresponding structure in the following table as the raw material, the corresponding example compounds in the table can be obtained through reductive amination reaction under the same conditions.

Example 91, the synthesis of compound 91

Referring to the method of Example 63, using the intermediate Z-31 and the SM-acid in the following table as raw materials, the compound 91 of Example can be obtained by condensation reaction under the same conditions.

Example 92, Example 93, Synthesis of Compound 92 and Compound 93

Referring to the method of Example 75, using the intermediate Z-32 as the raw material and the aldehyde of the corresponding structure in the following table as the raw material, the corresponding example compounds in the table can be obtained through reductive amination reaction under the same conditions.

Example 94, the synthesis of compound 94

Step 1. Synthesis of Intermediate 94-a

To Z-31 (36 mg, 67.83 μmol) in DCM (2 mL), add TEA to pH=9-10, then add Z-43 (41.89 mg, 81.40 μmol), stir at room temperature for 30 minutes, and then add acetic acid to adjust pH= 4-5. After stirring for 5 minutes, NaBH(OAc) ₃ (43.13 mg, 203.49 μmol) was added in one portion, and the reaction mixture was left at room temperature overnight. After the reaction was completed, water was added, DCM was added for extraction, the organic phase was washed with water, washed with brine, dried, concentrated, and the crude product was separated and purified by Pre-TLC (DCM/MeOH=10/1) to obtain intermediate 94-a (69 mg, 67.03 μmol, 98.82% yield). MS(ESI) m/z=1029[M+1] ⁺ .

Step 2. Synthesis of compound 94

At room temperature, TFA (18.04 mg, 158.25 μmol) was added to a solution of 94-a (70 mg, 68.00 μmol) in DCM (2 mL), and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated, and the crude product was treated with pre -HPLC separation and purification gave 94; (30.88 mg, 28.96 μmol, 42.58% yield, 95% purity, TF) as a yellow solid. MS (ESI) m/z = 899 [M+H] ⁺ . Nuclear magnetic spectrum: 1H NMR (400MHz, Methanol-d4)δ8.86(s,1H), 8.24(s,1H), 8.01(s,1H), 7.65–7.58(m,1H), 7.47–7.41(m ,3H),7.16(d,J=2.1Hz,1H),7.05(d,J=8.7Hz,1H),5.20–5.12(m,1H),4.56–4.44(m,3H),4.35–4.26( m, 1H), 4.16–4.08 (m, 1H), 3.95–3.86 (m, 2H), 3.84 (s, 3H), 3.79–3.67 (m, 2H), 3.57 (t, J=5.6Hz, 2H) ,3.46(t,J＝6.3Hz,2H),3.18-3.04(m,2H),2.96-2.80(m,2H),2.80(s,3H),2.73-2.66(m,2H),2.57-2.49 (m, 1H), 2.37–2.29 (m, 1H), 2.20–2.13 (m, 1H), 2.11–1.99 (m, 4H), 1.74–1.64 (m, 2H), 1.63–1.56 (m, 2H) ,1.46–1.37(m,4H),0.87(dd,J=6.6,3.2Hz,3H).

Synthesis of Example 95 to Example 101, Compound 95 to Compound 101

Referring to the method from step 1 to step 2 of Example 94, the SM-aldehyde and SM-amine in the following table are used as raw materials, and the corresponding example compounds in the table can be obtained by reductive amination.

In order to illustrate the beneficial effects of the present invention, the present invention provides the following test examples.

Test Example 1. Detection of EGFR Kinase Activity

KinEASE^TM试剂盒(CISBIO)中，具体操作如下：在384孔板中加入125pM EGFR(T790M/C797S/L858R)激酶蛋白，300nM TK Substrate-biotin，0.8μM ATP，反应总体积10μL，混匀后室温孵育40分钟；再加入37.5nM Streptavidin-XL665和5μL/孔TK Antibody-Cryptate，反应终体积20μL，混匀后室温孵育60分钟，用酶标仪检测激发波长320nM发射波长620nM和665nM的荧光强度，计算FRET信号强度(665nM荧光强度/620nM荧光强度*10000)。检测化合物对激酶活性抑制时，化合物先与激酶蛋白室温预孵育15～60分钟，再加入底物和ATP进行反应。活性数据如下表：EGFR kinase activity was characterized by detecting the phosphorylation level of the substrate, and the intensity of the FRET signal between the phosphorylated antibody and the biotin-labeled polypeptide substrate was proportional to the enzyme activity. The reagents required for the detection are included in the

KinEASE ^TM kit (CISBIO), the specific operation is as follows: add 125pM EGFR (T790M/C797S/L858R) kinase protein, 300nM TK Substrate-biotin, 0.8μM ATP to a 384-well plate, and mix at room temperature Incubate for 40 minutes; then add 37.5nM Streptavidin-XL665 and 5μL/well TK Antibody-Cryptate, the final reaction volume is 20μL, after mixing, incubate for 60 minutes at room temperature, and use a microplate reader to detect the fluorescence intensity of excitation wavelength 320nM emission wavelength 620nM and 665nM, Calculate the FRET signal intensity (665nM fluorescence intensity/620nM fluorescence intensity*10000). When detecting the inhibition of the kinase activity by the compound, the compound is pre-incubated with the kinase protein for 15-60 minutes at room temperature, and then the substrate and ATP are added for the reaction. The activity data are as follows:

Test example 2. Detection of EGFR-PROTAC-CRBN/DDB1 ternary complex formation

A series of concentrations of example compounds, EGFR (T790M/C797S/L858R) recombinant protein and CRBN/DDB1 recombinant protein complex were added to a 384-well plate. The reaction buffer was 1×PBS pH 7.4, incubated at room temperature for 120 minutes, and then 5nM Antibody was added. GST-Eu cryptate and 6.67nM, Ant 6His-XL665 were mixed and incubated at room temperature for 60 minutes, the fluorescence intensity of excitation wavelength 320nM emission wavelength 620nM and 665nM was detected by a microplate reader, and the FRET signal intensity was calculated (665nM fluorescence intensity/620nM fluorescence intensity *10000). The experimental results are shown in FIG. 1 , the compound of the present invention can form a ternary complex.

Test Example 3. Detection of EGFR degradation at the cellular level

96-well transparent bottom black plates were treated with collagen and fibronectin, and then seeded with Ba/F3-EGFR (T790M/C797S/L858R) overexpressing cells (2*10^4 cells per well) and incubated at 37°C for 5 Incubate overnight in a %CO ₂ incubator. The next day, remove the medium from the cells, add 150 μL of fresh medium first, then add 50 μL of medium containing 4 times the final concentration of the example compounds, put in an incubator, and continue at 37 °C and 5% _CO Incubate for 16 hours. After that, the medium in the cells was removed, and the cells were fixed with 4% paraformaldehyde, 150 μL per well, and incubated at room temperature for 15 min. After the cells were fixed, the cells were permeabilized with 0.1% (v/v) Triton X-100, 150 μL per well, and incubated at room temperature for 15 min. Cells were blocked with 1% blocking reagent (Roche, catalog number: 11096176001), 100 μL per well, and incubated at room temperature for 60 min. After blocking, 50 μL of EGFR antibody diluted with 1% (w/v) blocking reagent (CST, Cat. No.: 4267S, dilution ratio of 1:250) was added to each well, and incubated at room temperature for 2 hours. The EGFR antibody in the wells was removed, and the cells were washed with 0.05% (v/v) PBST, 200 μL per well, 4 times. Then add 1% (w/v) blocking reagent diluted goat anti-rabbit fluorescent secondary antibody (Invitrogen, Cat. No. A32731) and DAPI staining solution (Thermofisher, Cat. No. 62247) (fluorescent secondary antibody dilution ratio 1:250, DAPI The dilution ratio is 1:2000), 50 μL per well, and incubated at room temperature on a shaker at 300 rpm in the dark for 2 hours. Antibodies in the wells were then removed, and cells were washed with 0.05% (v/v) PBST, 200 μL per well, repeated 4 times (the last wash was retained in the wells). Data were acquired using a high-content imaging analysis system (GE, In CellAnalyzer, model: 2200), 9 fields of view were scanned per well, and the exposure time of the FITC channel was set to 0.5s, and the exposure time of the DAPI channel was set to 0.05s. The data were analyzed using the In cell analyzer workstation software to obtain intracellular EGFR fluorescence signal values and surrounding background values. Use Excel to perform subsequent processing on the acquired data. First, the intracellular EGFR fluorescence signal value is subtracted from the background value around the cell to obtain the normalized intracellular EGFR fluorescence signal value. Next, the relative levels of EGFR after compound treatment were calculated relative to the DMSO control (DMSO treatment counted as 100%), the relative levels of EGFR were analyzed using GraphPad Prism software, and DC ₅₀ values and fitted curves were obtained.

The experimental results show that the compound of the present invention can inhibit and degrade the EGFR protein, and may become a drug targeting the degradation of the EGFR protein.

Claims (13)

1. A bifunctional compound of formula I, or a stereoisomer, pharmaceutically acceptable salt thereof:

wherein,

R¹selected from hydrogen, halogen, ═ O, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R²selected from hydrogen, halogen, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

X¹、X²each independently selected from O, S, NR^x1、CR^x1R^x2；

R^x1、R^x2Each independently selected from hydrogen and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

y is selected from N or CR^Y1；

R^Y1Selected from hydrogen, C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

the ring A is selected from a benzene ring and a 5-6-membered aromatic heterocycle; wherein the benzene ringThe aromatic heterocyclic ring may be further substituted by one, two or three R^ASubstitution;

each R^AAre respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R⁴、R⁵0, 1,2 or 3 are respectively independent;

R⁴、R⁵are respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

Z¹、Z²、Z³、Z⁴each independently selected from CH or N;

the B ring is selected from 5-6 membered cycloalkyl, 5-6 membered heterocycloalkyl, 5-6 membered aromatic ring, 5-6 membered aromatic heterocycle; wherein the cycloalkyl, heterocycloalkyl, aryl, heteroaryl may be further substituted by one, two or three R^BSubstitution;

each R^BAre independently selected from hydrogen, halogen, cyano, nitro, ═ O, ═ S and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

each L^AAre respectively and independently selected from CRR, C (O), O, S, S (O), S (O)₂NR, -CR ═ CR-, -C.ident.C-, P (O) R, P (O) OR, 3-to 10-membered cycloalkane, 3-to 10-membered heterocycloalkane, 5-to 10-membered aromatic ring, 5-to 10-membered aromatic heterocycle, 5-to 12-membered spiro ring, 5-to 12-membered bridged heterocycle; wherein the cycloalkane, heterocycloalkane, aromatic ring, aromatic heterocycle, spiro ring, spiro heterocycle, bridged ring, bridged heterocycle may be further substituted with one, two or three R^LASubstitution;

each R^LAAre each independently selected from hydrogen, halogen, ═ O, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

each R is independently selected from hydrogen and C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl).

2. The bifunctional compound of claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein:

ring B is selected from

3. The bifunctional compound of claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein:

l is-L^A1-L^A2-L^A3-L^A4-L^A5-L^A6-L^A7-；

L^A1Is selected from- (C)_0～2Alkylene) -;

L^A2selected from-NR-, - (5-6 membered heterocycloalkyl) -or none;

L^A3selected from- (5-6 membered heterocycloalkyl) -, - (5-10 membered bridged heterocycle) -, - (6-9 membered spiroheterocycle) -, - (C)_1～10Alkylene) -NR-, - (5-to 6-membered heterocycloalkyl) -NR-, - (5-to 10-membered bridged heterocycle) -NR-, - (6-to 9-membered spiroheterocycle) -NR-, - (C)_0～10Alkylene) -;

L^A4selected from-C (O) -, - (5-6 membered heterocycloalkyl) -or none;

L^A5is selected from- (C)_0～10Alkylene) -or-c (o) -;

L^A6selected from-O-, -NR-, or none;

L^A7selected from the group consisting of- (CH ≡ CH) -, - (3-to 6-membered cycloalkane) -, - (3-to 6-membered heterocycle) -, - (5-to 6-membered heteroaromatic ring) -, - (6-to 9-membered spirocyclic) -, - (C-to 9-membered spiroheterocyclic ring) -, - (C-to 9-membered spirocyclic ring) -, and_0～10alkylene) -or none;

each R is independently selected from hydrogen and C_1～10An alkyl group.

4. The bifunctional compound of claim 3, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein:

L^A3is selected from

L^A7Is selected from- (CH ≡ CH) -, - (C)_0～10Alkylene) -, or,

Or none.

5. The bifunctional compound of claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein:

R¹is selected from methyl; r²Is selected from methyl; x¹Is selected from O; x²Is selected from CH₂(ii) a Y is selected from N;

ring A is selected from pyrazole or pyridine; wherein the pyrazole or pyridine may be further substituted by one, two or three R^ASubstitution; each R^AEach independently selected from hydrogen and methyl;

R⁴、R⁵each independently selected from hydrogen.

6. The bifunctional compound of claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein: the compound of formula I is represented by formula IIa:

wherein,

R¹selected from hydrogen, halogen, ═ O, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R²selected from hydrogen, halogen, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

X¹、X²each independently selected from O, S, NR^x1、CR^x1R^x2；

R^x1、R^x2Each independently selected from hydrogen and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

y is selected from N or CR^Y1；

R^Y1Selected from hydrogen, C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

the ring A is selected from a benzene ring and a 5-6-membered aromatic heterocycle; wherein the benzene ring and the aromatic heterocyclic ring can be further substituted by one, two or three R^ASubstitution;

each R^AAre respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R⁴、R⁵0, 1,2 or 3 are respectively independent;

R⁴、R⁵are respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

Z¹、Z²、Z³、Z⁴each independently selected from CH or N;

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

each L^AAre respectively and independently selected from CRR, C (O), O, S, S (O), S (O)₂NR, -CR ═ CR-, -C.ident.C-, P (O) R, P (O) OR, 3-to 10-membered cycloalkyl, 3-to 10-membered heterocycloalkyl, 5-to 10-membered aromatic ring, 5-to 10-membered aromatic heterocycle, 5-to 12-membered spiro ring, 5-to 12-membered bridged heterocycle; wherein the cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle, spiro heterocycle, bridged ring, bridged heterocycle may be further substituted with one, two or three R^LASubstitution;

each R^LAAre each independently selected from hydrogen, halogen, ═ O, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

each R is independently selected from hydrogen and C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl).

7. The bifunctional compound of claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein: the compound of formula I is represented by formula IIb:

wherein,

R¹selected from hydrogen, halogen, ═ O, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R²selected from hydrogen, halogen, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

X¹、X²each independently selected from O, S, NR^x1、CR^x1R^x2；

R^x1、R^x2Each independently selected from hydrogen and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

y is selected from N or CR^Y1；

R^Y1Selected from hydrogen, C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

the ring A is selected from a benzene ring and a 5-6-membered aromatic heterocycle; wherein the benzene ring and the aromatic heterocyclic ring can be further substituted by one, two or three R^ASubstitution;

each R^AAre respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R⁴、R⁵0, 1,2 or 3 are respectively independent;

R⁴、R⁵are respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

Z¹、Z²、Z³、Z⁴each independently selected from CH or N;

R^Bselected from hydrogen, halogen, cyano, nitro, ═ O, ═ S and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

each L^AAre respectively and independently selected from CRR, C (O), O, S, S (O), S (O)₂NR, -CR ═ CR-, -C.ident.C-, P (O) R, P (O) OR, 3-to 10-membered cycloalkyl, 3-to 10-membered heterocycloalkyl, 5-to 10-membered aromatic ring, 5-to 10-membered aromatic heterocycle, 5-to 12-membered spiro ring, 5-to 12-membered bridged heterocycle; wherein the cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle, spiro heterocycle, bridged ring, bridged heterocycle may be further substituted with one, two or three R^LASubstitution;

each R^LAAre each independently selected from hydrogen, halogen, ═ O, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

each R is independently selected from hydrogen and C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl).

8. The bifunctional compound of claim 1, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein: the compound of formula I is represented by formula IIc:

wherein,

R¹selected from hydrogen, halogen, ═ O, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R²selected from hydrogen, halogen, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

X¹、X²each independently selected from O, S, NR^x1、CR^x1R^x2；

R^x1、R^x2Each independently selected from hydrogen and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

y is selected from N or CR^Y1；

R^Y1Selected from hydrogen, C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

the ring A is selected from a benzene ring and a 5-6-membered aromatic heterocycle; wherein the benzene ring and the aromatic heterocyclic ring can be further substituted by one, two or three R^ASubstitution;

each R^AAre respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

R⁴、R⁵0, 1,2 or 3 are respectively independent;

R⁴、R⁵are respectively and independently selected from hydrogen, halogen, cyano, nitro and-C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

Z¹、Z²、Z³、Z⁴each independently selected from CH or N;

w is selected from C (O), CRR, NR;

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

each L^AAre respectively and independently selected from CRR, C (O), O, S, S (O), S (O)₂NR, -CR ═ CR-, -C.ident.C-, P (O) R, P (O) OR, 3-to 10-membered cycloalkyl, 3-to 10-membered heterocycloalkyl, 5-to 10-membered aromatic ring, 5-to 10-membered aromatic heterocycle, 5-to 12-membered spiro ring, 5-to 12-membered bridged heterocycle; wherein the cycloalkyl, heterocycloalkyl, aromatic ring, aromatic heterocycle, spiro heterocycle, bridged ring, bridged heterocycle may be further substituted with one, two or three R^LASubstitution;

each R^LAAre each independently selected from hydrogen, halogen, ═ O, cyano, nitro, -C_1～6Alkyl, halogen substituted-C_1～6Alkyl, -OR, -NRR, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl);

each R is independently selected from hydrogen and C_1～10Alkyl, halogen substituted-C_1～6Alkyl, -C_0～2Alkylene- (3-to 10-membered cycloalkyl), -C_0～2Alkylene- (3-to 10-membered heterocycloalkyl).

9. A bifunctional compound as claimed in claims 6-8, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein:

l is selected from the following groups:

-(C_0～2alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene) - (CH ≡ CH) -, C,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene group) - (C_0～10Alkylene) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene group) - (3-to 6-membered cycloalkane) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene group) - (3-to 6-membered heterocycle) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene group) - (5-to 6-membered aromatic heterocycle) -, or,

-(C_0～2Alkylene group) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene) compounds

-(C_0～2Alkylene group) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene group) - (5-to 6-membered aromatic heterocycle) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene group) - (5-to 6-membered aromatic heterocycle) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene) -O-),

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene) -O- (3-to 6-membered cycloalkane) -, and,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene) -O- (C)_0～10Alkylene) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～10Alkylene group) - (6-to 9-membered spiroheterocycle) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) -C (O) - (C)_0～10Alkylene) - (CH ≡ CH) -, C,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) -C (O) - (C)_0～10Alkylene) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) -C (O) - (C)_0～10Alkylene) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) - (C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) -C (O) - (C)_0～2Alkylene) -O-),

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) -C (O) - (C)_0～10Alkylene group) - (3-to 6-membered cycloalkane) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) -C (O) - (C)_0～10Alkylene group) - (3-to 6-membered heterocycle) -, or,

-(C_0～2Alkylene) - (5-to 6-membered heterocycloalkyl) -C (O) - (C)_0～10Alkylene) - (5-to 6-membered aromatic heterocycle) -.

10. The bifunctional compound of claim 9, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein: the method is characterized in that:

R¹is selected from methyl; r²Is selected from methyl; x¹Is selected from O; x²Is selected from CH₂(ii) a Y is selected from N;

ring A is selected from pyrazole or pyridine; wherein the pyrazole or pyridine may be further substituted by one, two or three R^ASubstitution; each R^AEach independently selected from hydrogen and methyl;

R⁴、R⁵each independently selected from hydrogen;

w is selected from C (O), CH₂、NH；

R^BIs selected from methyl;

l is selected from the following linking groups:

-(C_0～1alkylene) - (piperazine) - (C_0～10Alkylene) - (CH ≡ CH) -, C,

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) -, or,

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene group) - (C_0～10Alkylene) -, or,

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) -, or,

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) - (pyrazole) -,

-(C_0～1alkylene) - (piperazine) - (C_0～2Alkylene) - (piperazine) - (C_0～10Alkylene) - (pyrazole) -,

-(C_0～1alkylene) -piperazine-cyclobutane-s,

-(C_0～1Alkylene) - (piperazine) - (cyclobutylamine) -,

-(C_0～1alkylene) - (1.2.5.6-tetrahydropyridine) - (C)_0～10Alkylene) -, or,

-(C_0～1Alkylene) - (piperidine) - (C)_0～10Alkylene) -, or,

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) - (piperidine) -

-(C_0～1Alkylene) - (piperazine) -C (O) - (C)_0～10Alkylene) compounds

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) - (tetrahydropyrrole) -C (O) - (C)_0～10Alkylene) -O-),

-(C_0～1Alkylene) - (piperazine) -C (O) - (C)_0～10Alkylene) - (CH ≡ CH) -, C,

-(C_0～1Alkylene) - (piperazine) -C (O) - (C)_0～10Alkylene) -, or,

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) - (pyrazole) -,

-(C_0～1alkylene) - (piperazine) -C (O) - (C)_0～10Alkylene) - (pyrazole) -,

-(C_0～1alkylene) - (piperazine) - (C_0～10Alkylene) -O-),

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) -O- (C)_1～10Alkylene) s (a),

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) -O- (3-to 4-membered cycloalkane) -, and,

-(C_0～1Alkylene) - (piperazine) - (C_0～10Alkylene) - (2-azaspiro [ 3.3)]Heptane) -.

11. A bifunctional compound as claimed in claims 1-5, or a stereoisomer, pharmaceutically acceptable salt thereof, wherein: the compound is specifically:

12. use of the bifunctional compound of any one of claims 1-11, or a stereoisomer, a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment of a tumor or cancer.

13. A composition comprising a bifunctional compound of any one of claims 1-11, or a stereoisomer, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

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