CN114057770B

CN114057770B - Bifunctional compounds targeting EGFR protein degradation

Info

Publication number: CN114057770B
Application number: CN202110889913.3A
Authority: CN
Inventors: 李进; 潘飞; 陈迪; 兰燕; 汤若风; 辛艳飞; 张登友
Original assignee: Hitgen Inc
Current assignee: Hitgen Inc
Priority date: 2020-08-06
Filing date: 2021-08-06
Publication date: 2023-05-02
Anticipated expiration: 2041-08-06
Also published as: CN114057770A

Abstract

The invention provides a novel bifunctional compound, or a stereoisomer, a pharmaceutically acceptable salt and application thereof in preparing medicines. The bifunctional compound is a protein targeting complex, can selectively promote EGFR protein ubiquitination and degradation, and can be used for treating various cancers/tumors.

Description

Bifunctional compounds targeting EGFR protein degradation

Technical Field

The invention belongs to the field of medicines, and particularly relates to a bifunctional compound targeting EGFR protein degradation and application thereof in preparation of medicines.

Background

EGFR (Epidermal Growth Factor Receptor) is a member of the erbB receptor family of transmembrane protein tyrosine kinases. EGFR may form homodimers on cell membranes by binding to its ligand, e.g., epidermal Growth Factor (EGF), or heterodimers with other receptors in the family, such as erbB2, erbB3, or erbB 4. The formation of these dimers can cause phosphorylation of critical tyrosine residues within EGFR cells, thereby activating multiple downstream signaling pathways within the cell. These intracellular signaling pathways play an important role in cell proliferation, survival and anti-apoptosis. Deregulation of EGFR signaling pathways, including increased expression of ligands and receptors, EGFR gene amplification, and mutations, can promote transformation of cells into malignant tumors, and play an important role in proliferation, invasion, metastasis, and angiogenesis of tumor cells. EGFR is therefore considered as a target for the development of anticancer drugs.

Ubiquitin itself consists of 76 amino acid residues and has a molecular weight of about 8.5kDa, known as "Ubiquitin" because of its extensive presence in eukaryotic organisms. Ubiquitination (Ubiquitination) refers to the process of classifying proteins in cells under the action of a series of special enzymes, selecting target protein molecules from the proteins, and performing specific modification on the target proteins to form target protein polyubiquitin chains. These specific enzymes include ubiquitin activating enzyme (E1), ubiquitin binding enzyme (E2), ubiquitin ligase (E3), etc. Ubiquitination is closely related to the onset of diseases such as tumors, cardiovascular diseases, autoimmunity and the like. Ubiquitin-protease system degradation proteins are an important pathway for protein selective degradation and can be simply divided into three steps: firstly, a ligase is used for adding ubiquitin tags to target proteins, then a plurality of ubiquitin tags are obtained after a plurality of rounds of ubiquitination, and the proteins after polyubiquitination are identified by a proteasome and degraded. By constructing a bifunctional compound, one end is the structure of the target protein; the other end can be combined with a protein degradation system such as E3 ligase, and the middle is connected through a proper connecting chain, so that the degradation of target proteins can be effectively promoted.

EGFR inhibitor drugs that have been marketed mostly develop resistance to small molecule inhibitors after treatment. Its drug resistance mechanism includes EGFR secondary mutation, bypass activation, etc. At present, no drugs for degrading EGFR proteins by constructing bifunctional compounds are marketed. The development of a bifunctional compound capable of ubiquitinating and degrading EGFR protein can become a novel method for treating tumors or cancers, and has wide application prospect.

Disclosure of Invention

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

wherein, the liquid crystal display device comprises a liquid crystal display device,

R ¹ selected from hydrogen, halogen, =o, -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);

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

R ^x1 、R ^x2 Are respectively and independently selected from hydrogen and 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 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- (3-10 membered heterocycloalkyl);

the ring A is selected from benzene ring and 5-6 membered aromatic heterocycle; wherein the benzene ring, aromatic heterocycle may be further substituted with one, two or three R ^A Substitution;

each R ^A Independently 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);

R ⁴ 、R ⁵ 0, 1, 2 or 3 are respectively independent;

R ⁴ 、R ⁵ independently 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);

Z ¹ 、Z ² 、Z ³ 、Z ⁴ each 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 and 5-6 membered aromatic heterocycle; wherein the cycloalkyl, heterocycloalkyl, aromatic ring, or aromatic ring may be further substituted with one, two or three R ^B Substitution;

each R ^B 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 less than or equal to 100;

each L ^A Are respectively and independently selected from CRR, C (O), O, S, S (O) and S (O) ₂ NR, -CR=CR-, -C≡C-, P (O) R, P (O) OR, 3-10 membered cycloalkane, 3-10 membered heterocycloalkyl, 5-10 membered aromatic ring, 5-10 membered aromatic heterocycle, 5-12 membered spiro ring, 5-12 membered bridged heterocycle; wherein the cycloalkane, heterocycloalkyl, aromatic ring, aromatic heterocycle, spirocycle, spiroheterocycle, bridged ring, bridged heterocycle may be further substituted with one, two or three R ^LA Substitution;

each R ^LA 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- (3-10 membered heterocycloalkyl).

Further, the method comprises the steps of,

the B ring is selected from

Or->

Further, the method comprises the steps of,

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

Wherein L is ^A1 One end is provided with

Is connected with L at the other end ^A2 Are connected;

L ^A7 one end is connected with L ^A6 Is connected with the other end of

Are connected;

L ^A1 selected from- (C) _0～2 Alkylene) -;

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

L ^A3 Selected from- (5-6 membered heterocycloalkyl) -, - (5-10 membered bridged heterocyclic) -, - (6-9 membered spiro heterocyclic) -, - (C) _1～10 Alkylene) -NR-, - (5-to 6-membered heterocycloalkyl) -NR-, - (5-to 10-membered bridged heterocycle) -NR-, - (6-to 9-membered spiroheterocycle) -NR-, - (C _0～10 Alkylene) -;

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

L ^A5 selected from- (C) _0～10 Alkylene) -or-C (O) -;

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

L ^A7 selected from the group consisting of- (CH.ident.CH) -, - (3-to 6-membered cycloalkane) -, - (3-to 6-membered heterocyclic ring) -, - (5-to 6-membered aromatic heterocycle) -, - (6-to 9-membered spiro ring) -, - (C) _0～10 Alkylene) -or none;

each of which isR is independently selected from hydrogen, C _1～10 An alkyl group.

Still further, the method further comprises the steps of,

L ^A3 selected from the group consisting of

/>

L ^A7 Selected from- (CH≡CH) -, - (C) _0～10 Alkylene group) -,

or none.

Further, the method comprises the steps of,

R ¹ selected from methyl; r is R ² Selected from methyl; x is X ¹ Selected from O; x is X ² Selected from CH ₂ The method comprises the steps of carrying out a first treatment on the surface of the Y is selected from N;

ring a is selected from pyrazole or pyridine; wherein pyrazole or pyridine may be further substituted with one, two or three R ^A Substitution; each R ^A Each independently selected from hydrogen, methyl;

R ⁴ 、R ⁵ independently selected from hydrogen.

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

y is selected from N or CR ^Y1 ；

R ⁴ 、R ⁵ 0, 1, 2 or 3 are respectively independent;

Z ¹ 、Z ² 、Z ³ 、Z ⁴ Each independently selected from CH or N;

l is- (L) ^A ) _q -；

q is an integer greater than or equal to 1;

each L ^A Are respectively and independently selected from CRR, C (O), O, S, S (O) and S (O) ₂ NR, -CR=CR-, -C≡C-, P (O) R, P (O) OR, 3-10 membered ringAlkyl, 3-10 membered heterocycloalkyl, 5-10 membered aromatic ring, 5-10 membered aromatic heterocycle, 5-12 membered spiro ring, 5-12 membered bridged heterocycle; wherein cycloalkyl, heterocycloalkyl, aromatic ring, spiro ring, bridged ring may be further substituted with one, two or three R ^LA Substitution;

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- (3-10 membered heterocycloalkyl).

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

R ^x1 、R ^x2 Are respectively and independently selected from hydrogen and C _1～6 Alkyl, halogen substituted-C _1～6 Alkyl, -C _0～2 Alkylene- (3-to 10-membered cycloalkyl) radicals,-C _0～2 Alkylene- (3-10 membered heterocycloalkyl);

y is selected from N or CR ^Y1 ；

R ⁴ 、R ⁵ 0, 1, 2 or 3 are respectively independent;

Z ¹ 、Z ² 、Z ³ 、Z ⁴ each independently selected from CH or N;

R ^B 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 L ^A Are respectively and independently selected from CRR, C (O), O, S, S (O) and 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 ring,5-12 membered bridged ring, 5-12 membered bridged heterocycle; wherein cycloalkyl, heterocycloalkyl, aromatic ring, spiro ring, bridged ring may be further substituted with one, two or three R ^LA Substitution;

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

y is selected from N or CR ^Y1 ；

R ⁴ 、R ⁵ 0, 1, 2 or 3 are respectively independent;

Z ¹ 、Z ² 、Z ³ 、Z ⁴ each 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 ^A Are respectively and independently selected from CRR, C (O), O, S, S (O) and 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 ring, 5-12 membered bridged heterocycle; wherein cycloalkyl, heterocycloalkyl, aromatic ring, spiro ring, bridged ring may be further substituted with one, two or three R ^LA Substitution;

each R ^LA Independently selected from hydrogen, halogen, =o, cyano, nitro, -C _1～6 Alkyl, halogenPlain substituted-C _1～6 Alkyl, -OR, -NRR, -C _0～2 Alkylene- (3-10 membered cycloalkyl), -C _0～2 Alkylene- (3-10 membered heterocycloalkyl);

Further specifically, the method comprises the steps of,

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 group) -,

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

-(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) - (C _0～10 Alkylene radicals)

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

-(C _0～2 alkylene) - (5-6 membered heterocyclic ringAlkyl) - (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 group) -,

-(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 group) -,

-(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) -.

Still more particularly, the method comprises the steps of,

R ⁴ 、R ⁵ each independently selected from hydrogen;

w is selected from C (O), CH ₂ 、NH；

R ^B 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 group) -,

-(C _0～1 alkylene) - (piperazine) - (C _0～10 Alkylene) - (C _0～10 Alkylene group) -,

-(C _0～1 alkylene) - (piperazine) - (C _0～10 Alkylene group) -,

-(C _0～1 alkylene) - (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 group) -,

-(C _0～1 alkylene) - (piperidine) - (C _0～10 Alkylene group) -,

-(C _0～1 alkylene) - (piperazine) - (C _0～10 Alkylene) - (piperidine) compounds

-(C _0～1 Alkylene) - (piperazine) -C (O) - (C _0～10 Alkylene radicals)

-(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 group) -,

-(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 group),

-(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:

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1 end and the above structure

Are connected; the 2-terminal and +.>

Are connected.

In some embodiments of the invention, the compounds are specifically:

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the invention also provides application of the difunctional compound or stereoisomer and pharmaceutically acceptable salt thereof in preparing medicaments for treating tumors or cancers.

The invention also provides a composition comprising a bifunctional compound of any one of the above, or a stereoisomer, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

The compounds and derivatives provided in the present invention may be named according to IUPAC (international union of pure and applied chemistry) or CAS (chemical abstract service, columbus, OH) naming system.

Definition of terms used in connection with the present invention: unless otherwise indicated, the initial definitions provided for groups or terms herein apply to the groups or terms throughout the specification; for terms not specifically defined herein, the meanings that one skilled in the art can impart based on the disclosure and the context.

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

"further substituted" means that "substitution" may, but need not, occur, and that the description includes situations that may or may not occur.

The minimum and maximum values of the carbon atom content of the hydrocarbon groups are indicated by a prefix, e.g. prefix C _a～b Alkyl indicates any alkyl group containing from "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 indicated number of member atoms. For example, C ₁ ～ ₆ Alkyl refers to an alkyl group having 1 to 6 member atoms, for example 1 to 4 member atoms. The alkyl group may be linear or branched. Representative branched alkyl groups have one, two or three branches. The alkyl group 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) and hexyl. The alkyl group may also be part of another group, such as C ₁ ～ ₆ An alkoxy group.

"alkylene" as used herein refers to divalent saturated aliphatic hydrocarbon groups 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, "(C) ₁ ～C ₆ ) Alkylene "is intended to include methylene, ethylene, propylene, 2-methylpropylene, dimethylethylene, pentylene, and the like. Thus, the term "propylene" may be exemplified by the following structure:

likewise, the term "dimethylbutylene" may be used, for example, byAny of the examples of column structures: />

Or->

Furthermore, the term "(C) ₁ ～C ₆ ) Alkylene "is intended to include such branched chain hydrocarbon groups, such as cyclopropylmethylene, which may be exemplified by the following structures: />

"cycloalkyl", "cycloalkane" as used herein refers to a saturated or partially saturated cyclic group having multiple carbon atoms and no ring heteroatoms and having a single ring or multiple rings (including fused, bridged, spiro, and adamantane systems). For polycyclic systems having aromatic and non-aromatic rings that do not contain ring heteroatoms, the term "cycloalkyl" (e.g., 5,6,7,8, -tetrahydronaphthalen-5-yl) applies when the point of attachment is at a non-aromatic carbon atom. 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 comprising a multicycloalkyl ring system are dicyclohexyl, dicyclopentyl, bicyclooctyl, and the like. Two such bicycloalkyl polycyclic structures are exemplified and named below:

Dicyclohexyl and->

Dicyclohexyl group. Adamantyl groups include, but are not limited to, the following structures: />

"alkenyl" refers to a straight or branched hydrocarbon group 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 ethylenic unsaturation (> c=c <). For example, (Ca-Cb) alkenyl refers to an alkenyl group having a to b carbon atoms and is intended to include, for example, ethenyl, propenyl, isopropenyl, 1, 3-butadienyl, and the like.

"alkenylene" as used herein refers to a hydrocarbon chain having 2 to 10 carbon atoms, at least one double bond, and two unsaturated valences. For example, (C) ₃ -C ₆ ) Alkenylene group includes>C＝CH-CH ₂ -、-CH-CH＝CH-CH ₂ -and the like.

"alkynyl" refers to a straight or branched monovalent hydrocarbon radical 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.

"alkynylene" as used herein refers to divalent hydrocarbon chains having 2 to 10 carbon atoms and at least one triple bond.

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

"haloalkyl" means that a hydrogen atom in an alkyl group may be substituted with one or more halogen atoms. For example C _1～4 Halogen alkyl refers to an alkyl group containing 1 to 4 carbon atoms in which a hydrogen atom is substituted with one or more halogen atoms.

As used herein, "OR", "-NRR", etc. means that the R group is attached to the oxygen OR nitrogen atom by a single bond.

In the present invention, "-C (O) R", "-S (O) ₂ The oxygen atom in R' and the like is doubly bonded to a carbon atom or a sulfur atom.

"carbonyl" as used herein refers to an oxygen atom substituted by a double bond, i.e., "=o".

"heterocycle", "heterocycloalkyl", "heterocycloalkane" as used herein refers to a saturated or non-aromatic unsaturated ring containing at least one heteroatom; wherein the hetero atom means a nitrogen atom, an oxygen atom, a sulfur atom, etc. Typically represents a monovalent saturated or partially unsaturated mono-or bicyclic ring system of a plurality of ring atoms, preferably of 3 to 9 ring atoms, comprising 1, 2 or 3 groups selected fromN, O and S, the remaining ring atoms being carbon. Bicyclic means consisting of two rings sharing 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, azetidinyl, pyrrolidinyl, 2-oxo-pyrrolidin-3-yl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl,

Thiomorpholinyl, 1-dioxo-thiomorpholin-4-yl, azepanyl, diazepayl, homopiperazinyl or oxaazepanyl. An example of a bicyclic saturated heterocycloalkyl group is 8-aza-bicyclo [3.2.1]Octyl, quinuclidinyl, 8-oxa-3-aza-bicyclo [3.2.1]Octyl, 9-aza-bicyclo [3.3.1]Nonyl group,

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

As used herein, "aromatic ring", "aryl" refers to aromatic hydrocarbon groups having multiple carbon atoms. Aryl is typically a monocyclic, bicyclic or tricyclic aryl group having 5 to 20 carbon atoms. Furthermore, the term "aryl" as used herein refers to an aromatic substituent that may be a single aromatic ring or multiple aromatic rings fused together. Non-limiting examples include phenyl, naphthyl, or tetrahydronaphthyl.

"aromatic heterocycle", "aromatic heterocyclic group" as used herein refers to an aromatic unsaturated ring containing at least one heteroatom; wherein the hetero atom means a nitrogen atom, an oxygen atom, a sulfur atom, etc. An aromatic mono-or bicyclic hydrocarbon typically comprising a plurality of ring atoms, wherein one or more of the ring atoms is selected from heteroatoms of O, N, S. Preferably one to three heteroatoms. Heteroaryl represents, for example: pyridyl, indolyl, quinoxalinyl, quinolinyl, isoquinolinyl, benzothienyl, benzofuranyl, benzothienyl, benzopyranyl, benzothiopyranyl, furanyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, oxadiazolyl, benzimidazolyl, benzothiazolyl, benzoxazolyl.

"stereoisomers" include enantiomers and diastereomers;

the term "pharmaceutically acceptable" means that the carrier, cargo, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients comprising the pharmaceutical dosage form, and physiologically compatible with the recipient.

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

It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

Drawings

FIG. 1 is a graph showing the fluorescence detection result of EGFR-PROTAC-CRBN/DDB1 ternary complex formed by the compound of the invention.

Detailed Description

The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.

The known starting materials of the present invention may be synthesized using or according to methods known in the art, or may be purchased from An Naiji chemical, chengkoulochemical, shaoshan chemical technology, carbofuran technology, and the like.

The reagents described in the examples are abbreviated as follows: AIBN: azodiisobutyl cyanide; NBS: n-bromosuccinimide; DMP: DESS-MARTIN PERIODINANE DESS-martin oxidizer; DIPEA: n, N-diisopropylethylamine; TEMPO:2, 6-tetramethylpiperidine oxide; MTBE: methyl tertiary butyl ether; naHMDS: sodium bis (trimethylsilyl) amide; DIAD: diisopropyl azodicarboxylate; DPPA: diphenyl azide phosphate; TBTU:2- (1H-benzotrisazo L-1-yl) -1, 3-tetramethylurea tetrafluoroborate; 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.

The reaction was carried out under nitrogen atmosphere without specific explanation in examples. The examples are not specifically described, and the solution refers to an aqueous solution. The temperature of the reaction was room temperature, unless otherwise specified in the examples. The room temperature is the most suitable reaction temperature and is 20-30 ℃. In the examples, M is mol/liter unless otherwise specified.

The structure of the compounds was determined by Nuclear Magnetic Resonance (NMR) and Mass Spectrometry (MS). NMR shift (. Delta.) is given in units of 10-6 (ppm). NMR was performed using a nuclear magnetic resonance apparatus (Bruker Avance III 400 and Bruker Avance 300) with deuterated dimethyl sulfoxide (DMSO-d) ₆ ) Deuterated chloroform (CDCl) ₃ ) Deuterated methanol (CD) ₃ OD), internal standard is Tetramethylsilane (TMS). LC-MS was measured using Shimadzu LC-MS 2020 (ESI). HPLC was performed using a Shimadzu high pressure liquid chromatograph (Shimadzu LC-20A). MPLC (Medium pressure preparative chromatography)) The reverse phase preparative chromatograph was used with Gilson GX-281. The specification of the thin layer chromatography separation and purification product adopted by the smoke table yellow sea HSGF254 or Qingdao GF254 silica gel plate is 0.4 mm-0.5 mm. Column chromatography generally uses tobacco stand yellow sea silica gel 200-300 mesh silica gel as a 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) were added AIBN (3.66 g,22.26 mmol) and NBS (47.55 g,267.17 mmol). The reaction mixture was heated to 85℃and reacted for 4 hours. After completion of the reaction, the reaction solution was concentrated to give 1-a (65 g,211.06mmol,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,163.65mmol,28.51 mL), the reaction solution was warmed to 90℃and stirred at this temperature for 3 hours, after completion of the reaction, the mixture was concentrated to give a crude product, which was washed with water and filtered to give compound 1-b (35.6 g,110.17mmol,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) was added successively 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. Mu. Mol) and DIPEA (2.82 g,27.85mmol,3.88 mL). The reaction solution is N ₂ Stirring at 80deg.C for 18 hr under protection, diluting with water, extracting with ethyl acetate, washing the separated organic phase with water and saline, drying, and spin drying. Ethyl acetate was added to the crude product, a brown solid precipitated, and a filter cake was collected after filtration to give the objective compound 1-c (1.6 g,4.51mmol,48.65% yield). MS (ESI) m/z=355 [ m+1 ] ] ⁺ 。

Step 4, synthesis of 1-d

To a mixture 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, the mixture was stirred at room temperature for 18 hours under the protection of hydrogen. After completion of the reaction, pd/C was removed by filtration, and the filtrate was concentrated to give 1-d (162 mg,0.49mmol,80% yield, crude).

Step 5, synthesis of intermediate Z-1

To a mixture of compound 1-d (300 mg, 881.39. Mu. Mol) in DCM (3 mL) and DMF (3 mL) was added dessmartin reagent (560.75 mg,1.32 mmol). The suspension was stirred at room temperature for 1 hour. After the completion of the reaction, the mixture was filtered, the filtrate was concentrated to dryness, the crude product was diluted with ethyl acetate, a brown solid was precipitated, and the solid was collected by filtration and purified by reverse phase C18 column to give 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 the steps 2 to 5 in the synthesis method of the intermediate Z-1, the 2-bromomethyl-4-bromobenzoate used in the step 1 is taken as a raw material, and the rest conditions are unchanged, so that Z-2 is obtained. MS (ESI) m/z=339 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-3

According to steps 1 to 5 in the synthesis method of the intermediate Z-1, replacing the methyl 2-methyl-3-bromobenzoate in the step 1 with methyl 2-methyl-5-bromobenzoate, and obtaining Z-3 under the same conditions. MS (ESI) m/z=339 [ m+1 ] ] ⁺ 。

Synthesis of intermediates Z-4 to Z-12

According to steps 1 to 5 in the synthesis method of the intermediate Z-1, the methyl 2-methyl-3-bromobenzoate in the step 1 is replaced by SM-Br in the table, the 4-pentyne-1-alcohol in the step 3 is replaced by corresponding SM-ene in the table, and the rest conditions are unchanged, so that the structure of the corresponding Z-4 collocation Z-12 in the table is obtained.

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Synthesis of intermediate Z-13

To a mixture of intermediate compounds 1-c (500 mg,1.53 mmol) in DMF (4 mL)/DCM (40 mL) was added DMP (974.75 mg,2.30 mmol) at zero temperature, the reaction mixture was slowly warmed to room temperature and stirred for 3 hours, after completion of the reaction, 1mL of methanol was added and the reaction mixture was dried by spin-drying to give intermediate compound Z-13 (133 mg, 410.08. Mu. Mol,26.77% yield) from the residue by separation and purification by m-HPLC. MS (ESI) m/z=325 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-14

The synthesis method of the reference compound Z-13 is characterized in that the raw material 1-c is replaced by 2-c, and the rest methods are unchanged, so that Z-14 is obtained. MS (ESI) m/z=325 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-15

The synthesis method of the reference compound Z-13 is characterized in that the raw material 1-c is replaced by 3-c, and the rest methods are unchanged, so that Z-15 is obtained. 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 materials 1-c are replaced by corresponding raw material intermediate numbers, and the rest methods are unchanged, so that the corresponding Z-16 to Z-27 in the table are obtained.

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Synthesis of intermediate Z-25

To a mixture of 1-c (1 g,3.06 mmol) in acetonitrile (66.67 mL)/water (133.33 mL) was added TEMPO (191.51 mg,1.23 mmol) and iodobenzene diacetic acid (3.95 g,12.26 mmol). The reaction mixture was stirred at room temperature overnight, after completion of the reaction, the reaction was concentrated and the crude product was purified by normal phase silica gel column (DCM: meoh=33:1) to give 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 completion of the reaction, pd/C was filtered off, and the filtrate was dried by spinning to obtain 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 warmed to 115℃and reacted overnight. After completion of the reaction, the reaction mixture was cooled to room temperature, 7g of KOH was added, the mixture was separated, the separated aqueous phase was washed with MTBE 2 times, then pH was adjusted with concentrated hydrochloric acid=2-3, ethyl acetate was extracted, and the obtained organic phase was dried and concentrated to give intermediate compound 27-a (5.4 g,25.93mmol,25.96% yield), MS (ESI) m/z=207 [ M+1 ] ⁺

Step 2, synthesis of 27-b

To a solution of 27-a (2 g,9.60 mmol) in DCM (100 mL) at zero degrees was added 1 drop of DMF followed by slow dropwise addition (COCl) ₂ (2.44 g,19.21 mmol) was allowed to react at room temperature for one hour, and after completion of the reaction, the reaction solution was concentrated to give compound 27-b (2.1 g,9.26mmol,96.46% yield).

Step 3, synthesis of 27-c

(R) -4-benzyl-2-oxazolidinone (5.77 g,32.55 mmol) was dissolved in THF (80 mL), cooled to-78deg.C under nitrogen protection, n-BuLi (16 mL) was added, the reaction was allowed to proceed for 30 min at the temperature, then 27-b (7.38 g,32.55 mmol) in THF was added, the dropwise addition was completed, and the reaction was allowed to proceed for 1 hr. After the reaction is completed, the reaction is poured into saturated ammonium chloride aqueous solution to be quenched and separated,

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

Step 4, synthesis of 27-d

To 27-c (7.04 g,19.16 mmol) of THF (50 mL) under nitrogen protection at-78 ℃ was added dropwise NaHMDS (12.5 mL), the reaction was continued for 30 minutes while keeping the temperature, then MeI (16.32 g,114.96 mmol) of THF solution was added dropwise, the external bath was removed after the addition, the reaction was allowed to react at room temperature for 1 hour, after completion of the reaction, the reaction solution was poured into a saturated aqueous ammonium chloride solution to quench, the separated solution was concentrated, and the residue was purified by normal phase silica gel column separation (MTBE/PE=1/1) to give 27-d (5.4 g,14.16mmol,73.88% yield). MS (ESI) m/z=382 [ m+1 ] ] ⁺

Step 5, synthesis of 27-e

To 27-d (5.4 g,14.16 mmol) in THF (100 mL) at zero degree, liAlH was added ₄ (15 mL) was allowed to react at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated to dryness, and the residue was purified by normal phase silica gel column (MTBE/pe=1/2) to give 27-e (2.4 g,11.52mmol,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) and PPh was added sequentially under nitrogen ₃ (6.04 g,23.04 mmol), DIAD (4.65 g,23.04 mmol) and DPPA (6.34 g,23.04 mmol) were stirred overnight at room temperature, after the reaction was completed, the reaction mixture was concentrated to dryness and the residue was purified by normal phase silica gel column separation (8% MTBE in PE) to give 27-f (1.97 g,8.44mmol,73.28% yield). MS (ESI) m/z=234 [ m+1 ]] ⁺

Step 7, synthesis of 27-g

To 27-f (2 g,8.57 mmol) of MeOH (50 mL) was added 2 drops of concentrated hydrochloric acid followed by Pd/C (0.5 g). After hydrogen substitution, stirring was carried out at room temperature for 2 hours. After completion of the reaction, pd/C was filtered off and the filtrate was concentrated to dryness to give 27-g (1 g,8.53mmol,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 sequentially ₂ CO ₃ (2.36 g,17.07 mmol) and 2-fluoro-4-bromonitrobenzene (1.88 g,8.53 mmol), warmed to 60 ℃ and stirred overnight, after completion of the reaction, the reaction mixture was concentrated and the residue was purified by normal phase silica gel column (PE/ea=5/1) to give 27-h (2 g,6.31mmol,73.89% yield). MS (ESI) m/z=318 [ m+1 ] ] ⁺

Step 9, synthesis of 27-i

To a solution of 27-h (463mg, 1.47 mmol) in DCM (10 mL) at zero temperature were added TEA (222.52 mg,2.20mmol, 306.72. Mu.L) and MsCl (201.52 mg,1.76 mmol) in sequence, stirred for 1 hour at zero temperature, after completion of the reaction, saturated aqueous sodium bicarbonate solution was added, the separated organic phase was separated in succession by water washing, saturated brine washing, dried over anhydrous sodium sulfate, concentrated to give 27-i (0.57 g,1.44mmol,98.36% yield) and the crude product was directly reacted in the next step without purificationShould be. 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-pyrazol-4-yl) -6-methyliisonicotinic acid ester (362.18 mg,1.46 mmol) and K ₂ CO ₃ (505.37 mg,3.66 mmol) was dissolved in DMF (15 mL), the reaction was allowed to warm to 60℃and stirred overnight, after completion of the reaction, cooled to room temperature, MTBE was added, the mixture was washed 2 times with water, the separated organic phase was concentrated, and the residue was purified by column chromatography over silica gel (60% EA in PE) to give 27-j (254 mg, 464.85. Mu. Mol,31.73% yield). MS (ESI) m/z=548 [ m+1 ]] ⁺

Step 11, synthesis of 27-k

To 27-j (254 mg, 464.85. Mu. Mol) in MeOH (10 mL) at zero, NH was added in sequence ₄ OH (3 mL) and Na ₂ S ₂ O ₄ (404.42 mg,2.32 mmol) for 1 hour at room temperature, after the completion of the reaction, water was added to wash the stone, extracted with ethyl acetate, and the combined organic phases were washed successively with water and saturated brine, dried, and concentrated to give 27-k (170 mg, 329.18. Mu. Mol,70.81% yield), the crude product was used directly in the next reaction without purification. MS (ESI) m/z=518 [ m+1 ] ] ⁺

Step 12, synthesis of 27-l

To a solution of 27-k (170 mg, 329.18. Mu. Mol) in DCM (10 mL)/t-BuOH (3 mL) was added CNBr (41.87 mg, 395.02. Mu. Mol) and stirred overnight at room temperature. After completion of the reaction, a saturated aqueous sodium hydrogencarbonate solution was added, stirred for 10 minutes, and the organic phase was separated, washed with a saturated aqueous sodium hydrogencarbonate solution, water, brine in this order, dried over anhydrous sodium sulfate, and concentrated to give 27-l (177 mg, 326.91. Mu. Mol,99.31% yield). MS (ESI) m/z=541 [ m+1 ]] ⁺

Step 13, synthesis of 27-m

To 27-l (177 mg, 326.91. Mu. Mol) of THF (5 mL)/H ₂ To O (5 mL), naOH (52.31 mg,1.31 mmol) was added, and after completion of the reaction, the reaction mixture was concentrated, diluted with water, adjusted to pH=4-5 with 1M HCl, solid was separated out, filtered, and the cake was washed with water and dried to give 27-M (172 mg, 326.12. Mu. Mol,99.76% yield). MS (ESI) m/z=541 [ m+1 ]] ⁺

Step 14, synthesis of Z-27

To 27-m (172 mg, 326.12. Mu. Mol) of DCM (5 mL) was added TEA (132.00 mg,1.30mmol, 181.94. Mu.L) and TBTU (100.14 mg, 391.34. Mu. Mol) in this order, and after completion of the reaction, water was added for dilution, the organic phase was separated, concentrated, and the resulting residue was purified by silica gel column separation (DCM/MeOH=10/1) to give intermediate Z-27 (113 mg, 221.83. Mu. Mol,68.02% yield). MS (ESI) m/z=511 [ m+1 ] ] ⁺ ， ¹ H NMR(400MHz,DMSO-d ₆ )δ12.83(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. Mu. Mol) in DMF (30 mL) was added DIPEA (375.24 mg,2.90mmol, 505.71. Mu.L) and SEMCl (242.44 mg,1.45 mmol), the reaction mixture was warmed to stirring overnight at 60℃and after completion of the reaction, the reaction mixture was concentrated, the residue was diluted with water, extracted with ethyl acetate, the combined organic phases were washed successively with water, saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by normal phase silica gel column separation (100% EA) to give 28-a (373 mg, 583.13. Mu. Mol,60.25% yield). MS (ESI) m/z=639 [ m+1 ]] ⁺ 。

Step 2, synthesis of 28-b

To a mixed solution of 28-a (428 mg,1.12 mmol) in dioxane (20 mL)/water (2 mL) was added 4-N-Boc piperazine 1-methyl boron trifluoride potassium salt (515.51 mg,1.68 mmol), X-Phos (106.86 mg, 224.50. Mu. Mol), pd (OAc) ₂ (25.14 mg, 112.25. Mu. Mol) and Cs ₂ CO ₃ (1.10 g,3.37 mmol) was replaced with nitrogen several times, then warmed to 100℃under nitrogen and stirred for 15h. After the reaction is completedThe reaction solution was concentrated, the residue was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and the concentrated residue was purified by normal phase silica gel column separation (100% EA) to give 28-b (603 mg, 799.71. Mu. Mol,71.25% yield). MS (ESI) m/z=759 [ m+1 ] ] ⁺ ， ¹ H NMR(400MHz,DMSO-d ₆ )δ12.83(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. Mu. Mol) of DCM (5 mL) was added TFA (5 mL), and the reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was concentrated to give Z-28 (452 mg, 855.01. Mu. Mol,96.72% yield) which was 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 2-fluoro-4-bromonitrobenzene is replaced by 2-fluoro-5-bromonitrobenzene in step 8, and Z-29 can be obtained by the same method. MS (ESI) m/z=509 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-30

Referring to the synthesis method of Z-28, Z-27 in step 1 is replaced by Z-29, and the rest method steps are the same, so that Z-30 is obtained. 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. Mu. Mol) in DMF (30 mL) was added DIPEA (375.24 mg,2.90mmol, 505.71. Mu.L) and SEMCl (242.44 mg,1.45 mmol) in this order, the reaction mixture was warmed to 60℃and stirred for 12 hours, after completion of the reaction, the reaction mixture was concentrated, the residue was diluted with water and extracted with ethyl acetate, the combined organic phases were successively washed with water, saturated brine, dried over anhydrous sodium sulfate, and purified by separation on a normal phase silica gel column (100% ethyl acetate) after spin-drying to give 31-a (373 mg, 583.13. Mu. Mol,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. Mu. Mol) in toluene (3 mL) was added 1-t-butoxycarbonylpiperazine (14.56 mg, 78.17. Mu. Mol), BINAP (9.73 mg, 15.63. Mu. Mol), pd (OAc) ₂ (1.75 mg, 7.82. Mu. Mol) and Cs ₂ CO ₃ (50.81 mg, 156.33. Mu. Mol). The temperature was raised to 120℃under nitrogen and the reaction was stirred at this temperature for 15 hours. After the completion of the reaction, the mixture was diluted with ethyl acetate, washed with water, and the organic phase was concentrated to give a crude product, which was purified by reverse phase silica gel column to give 31-b (30 mg, 40.27. Mu. Mol,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. Mu. Mol) in DCM (3 mL) was added TFA (2 mL) and the mixture was stirred at room temperature for 1 hour. The reaction was then concentrated and the residual TFA was removed by oil pump to give Z-31 (50 mg,43.8% purity) which was 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 Z-31 synthesis method, Z-29 is used to replace Z-27 in step 1, and Z-32 is obtained by the same method. MS (ESI) m/z=515 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-33

Step 1, synthesis of intermediate 33-a

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

Step 2, synthesis of intermediate 33-b

Ice bath and nitrogen protection to 3-bromopiperidine-2, 6-dione (10 g,52.08 mmol), 4-methoxybenzyl alcohol (7.92 g,57.29 mmol) and PPh ₃ To a mixed solution of (13.66 g,52.08 mmol) was added dropwise DEAD (9.06 g,52.08 mmol), the reaction mixture was stirred at zero degree for 2 hours, after completion of the reaction, the mixture was quenched with water, extracted with ethyl acetate, and the combined organic phases were concentrated to give crude product, which was purified by silica gel column separation (PE/ea=3/1 and DCM/pe=1/1, v/v) to give intermediate 33-b (4.88 g,15.63mmol,30.02% yield). MS (ESI) m/z=312, 314[ m+1 ]] ⁺ 。

Step 3, synthesis of intermediate 33-c

A solution of 33-a (4.0 g,17.7 mmol) in THF (50 mL) was cooled to 0deg.C, t-BuOK (2.38 g,21.2 mmol) was added and the mixture stirred for 30 min at zero degrees, then-b (8.26 g,26.5 mmol) was added and the mixture stirred overnight at room temperature, the solvent was removed by spinning, and the crude product was purified by column chromatography over silica gel (EtOAc: DCM=1:1) to give 33-c (2.9 g,6.33mmol, 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) at room temperature was added MeSO ₃ H (10 mL), the reaction mixture was warmed to 110℃and stirred at this temperature for 2H. Cooling to room temperature, concentrating the mixture under reduced pressure, and coarse The product was diluted with ice water, the precipitated solid was collected by filtration, then washed with n-hexane, and dried to give 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 synthesis method of the intermediate Z-33 in the step 1, 33-a is obtained by replacing the raw material 4-bromo-2-methylaminoaniline with 5-bromo-2-methylaminoaniline, and the rest of the operations are unchanged. 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 at zero degrees ₂ (25 g,207 mmol). The mixture was warmed to 85 ℃ and stirred for 3 hours, then the reaction was cooled to room temperature and stirring was continued for 6 hours. After the raw materials are reacted completely, the mixed solution is concentrated under reduced pressure. Under nitrogen, the crude product was dissolved in DCM dried at zero degrees and Et was then added ₃ N (18.7 g,184 mmol) and 4-methoxybenzylamine (10.1 g,74 mmol), the mixture was stirred at room temperature for 3 hours. After completion of dilution with water, the mixture was extracted with DCM, the combined organic phases were washed successively with 0.5M HCl (50 mL) in water and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and spun-dried, and the crude product was purified by column chromatography on silica gel to give 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) of anhydrous THF (120 mL) was cooled to-78deg.C, then t-BuOK (5.4 g,48.4 mmol) of anhydrous THF (100 mL) was added dropwise under nitrogen protection at-78deg.C, the reaction mixture was stirred at-40deg.C for 1 hour, and after completion of the reaction, the mixture was saturated with NH ₄ Cl solution quenching, ethyl acetate extraction, combined organic phase with saturated brine washing, anhydrous sodium sulfate drying, filtering, concentration, vinegar bottle with silica gel column separation purification to obtain intermediate 34-c (10.9 g, 43.7)3mmol,77.7% yield). MS (ESI) m/z=250.0 [ m+1 ]] ⁺ 。

Step 4, synthesis of intermediate 34-d

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 was added dropwise trifluoromethanesulfonic anhydride (18.51 g,65.59 mmol) and the mixture stirred at 0-10deg.C under nitrogen for 1.5 h. After completion of the reaction, the mixture was concentrated, and the crude product was purified by silica gel column to give 34-d (8.17 g,21.43mmol,49% yield), MS (ESI) m/z=382 [ m+1 ]] ⁺ 。

Step 5, synthesis of intermediate 34-e

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

Step 6, synthesis of intermediate Z-34

Referring to step 4 of intermediate Z-33, substrate 33-c in step was replaced with 34-e, and the remaining steps were identical 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 of step 1 to step 4, in which step 1 3-bromo-2-methylaminoaniline is used instead of 4-bromo-2-methylaminoaniline as starting material, the rest of the operations are the same, and the same steps are performed to obtain intermediate Z-35, MS (ESI) m/z=338, 340[ M+1 ]] ⁺ 。

Synthesis of intermediate Z-36

Step 1, synthesis of intermediate 36-a

4-Pyrazoleboronic acid pinacol ester (5 g,25.77 mmol) and 3-benzyloxy bromopropane (8.86 g,38.65 mmol) were dissolved in DMF (20 mL) and then K was added ₂ CO ₃ (8.90 g,64.42 mmol), KI (427.76 mg,2.58 mmol). The reaction mixture was warmed to 80 ℃ and stirred for 17 hours. After completion of the reaction, the crude product obtained by the reaction mixture was concentrated and purified by silica gel column (PE: ea=8:1, v/v) to obtain intermediate 36-a (6.4 g,18.70mmol,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 dissolved in DMF (20 mL) and Na was added ₂ CO ₃ (1.86 g,17.53 mmol) and Pd (dppf) Cl ₂ (213.80 mg,292.19 umol) the mixture was replaced with nitrogen several times, then warmed to 100℃under nitrogen and stirred overnight. After completion of the reaction, 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 purified by column chromatography on silica gel (100% ea) to give intermediate 36-b (1.6 g,3.49mmol,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 stirred at room temperature under hydrogen protection for 24 hours. After completion of the reaction, pd/C was filtered off, and the filtrate was concentrated to give 36-C (367 mg, 996.24. Mu. Mol,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. Mu. Mol) in DCM (30 mL) was added DMP (230.19 mg, 542.91. Mu. Mol) and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated, and the obtained crude product was 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 4-pyrazoloboronic acid pinacol ester (6.0 g,30.92 mmol) in acetonitrile (60 ml) were added CsF (470 mg,3.09 mmol) and tert-butyl acrylate (5.82 mL,40.2 mmol). The reaction mixture was warmed to 80 ℃ and stirred at this temperature overnight. After the completion of the reaction, cooled to room temperature, concentrated under reduced pressure, 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, and the obtained crude product was purified by separation on a silica gel column (n-hexane/ethyl acetate=1/1, v/v) to give 37-a (9.37 g,29.07mmol, yield 94%). MS (ESI) m/z=323 [ m+h ]] ⁺ 。

Step 2, synthesis of intermediate 37-b

To 37-a (500 mg,1.55 mmol) and 1-b (455.86 mg,1.41 mmol) of DMF (12 mL)/H ₂ K was added to O (3 mL) solution ₃ PO ₄ (416.73 mg,1.98 mmol), S-phos (69.41 mg, 169.29. Mu. Mol) and Pd (OAc) ₂ (22.61 mg, 98.75. Mu. Mol), the mixture was heated to 100℃under the protection of nitrogen after nitrogen substitution, and stirred at this temperature for 15 hours, and the reaction was completed. Cooled to room temperature, filtered, the filtrate 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, concentrated, and the crude product was purified by column chromatography on silica gel (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. Mu. Mol) in DCM (2 mL) at room temperature was added TFA (1.54 g,13.51mmol,1 mL) and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated and dried to give Z-37 (92 mg, 185.34. Mu. Mol,81.27% yield), which was 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 CCl of methyl 2-methyl 5-iodobenzoate (10 g,36.22 mmol) ₄ To a solution of (100 mL) was added NBS (6.77 g,38.03 mmol) and benzoyl peroxide (4.38 g,18.11 mmol), and the mixture was warmed to 80℃and stirred for 4 hours. After completion of the reaction, cooled to room temperature, filtered, and the filtrate was concentrated to give 38-a (12 g,33.81mmol,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 K ₂ CO ₃ (14.02 g,101.42 mmol). The reaction mixture was warmed to 70 ℃ and stirred for 18h. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water, stirred for 10 minutes, filtered, and the cake was washed with petroleum ether and ethyl acetate, and the cake was collected and dried to give 38-b (6.5 g,17.56mmol,51.95% yield.) MS (ESI) m/z=371 [ m+h ] ] ⁺ 。

Step 3, synthesis of intermediate 38-c

To a solution of 38-b (400 mg,1.08 mmol) and tert-butyl 2- (4, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazol-1-yl) 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. Mu. Mol) and Pd (OAc) ₂ (17.32 mg, 75.65. Mu. Mol) was replaced with nitrogen several times, and then warmed to 100℃under nitrogen and stirred overnight. After the completion of the reaction, cooled to room temperature, diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a crude product which was taken up in DMF/H ₂ The O mixture was recrystallized to give 38-c (200 mg, 471.20. Mu. Mol,43.60% yield) 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. Mu. Mol) in DCM (3 mL) at room temperature was added TFA (53.73 mg, 471.20. Mu. Mol, 35.12. Mu.L) and the reaction mixture was stirred at room temperature for 2 hours. After the completion of the reaction, the reaction mixture was concentrated, and the crude product was dried to give Z-38 (200 mg, 414.62. Mu. Mol,87.99% yield, TF) which was used in the next reaction without further purification.

Synthesis of intermediate Z-39

Step 1, synthesis of intermediate 39-a

To intermediate 38-b (1 g,2.70 mmol) and 36-a (1.02 g,2.97 mmol) in dioxane (20 mL)/H ₂ Na was added to the mixture of O (1 mL) ₂ CO ₃ (400.93 mg,3.78 mmol), S-phos (132.92 mg, 324.20. Mu. Mol) and Pd (OAc) ₂ (43.31 mg, 189.12. Mu. Mol), the mixture was replaced with nitrogen several times, the temperature of the reaction mixture was raised to 60℃under the protection of nitrogen, and the reaction was stirred at 60℃overnight. After the completion of the reaction, the reaction solution was filtered, the filtrate was concentrated, the resulting 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 after spin-drying, the crude product was isolated and purified by pre-HPLC to give intermediate 39-a (130 mg, 283.53. Mu. Mol,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. Mu. Mol,10% purity) was added to a mixture of 39-a (410 mg, 894.20. Mu. Mol) in MeOH (150 mL) and THF (30 mL), the reaction was replaced with hydrogen several times, then stirred at room temperature under the protection of hydrogen for 15 hours, after completion of the reaction, filtered, and the filtrate was concentrated and dried to give a crude intermediate 39-b (310 mg) which was used in the next reaction without purification.

Step 3, synthesis of intermediate Z-39

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

Synthesis of intermediate Z-40

Step 1, synthesis of intermediate 40-a

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

Step 2, synthesis of intermediate 40-b

To 40-a (1 g,2.21 mmol) and 36-a (830.31 mg,2.43 mmol) dioxane (5.00 mL)/H ₂ Na was added to the O (5.00 mL) mixture ₂ CO ₃ (327.30 mg,3.09 mmol), S-phos (108.51 mg, 264.67. Mu. Mol) and Pd (OAc) ₂ (35.35 mg, 154.39. Mu. Mol) was replaced with nitrogen gas several times, then heated to 60℃under nitrogen gas protection, and stirred at this temperature for 15 hours, 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 phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, and the crude product was purified by silica gel column separation (PE/EA=1/1) after spinning to obtain intermediate 40-b (200 mg, 339.69. Mu. Mol,15.40% yield) as a white solid. MS (ESI) m/z=589 [ m+h ] ] ⁺ 。

Step 3 and step 4, synthesis of intermediate Z-40

Referring to the method of step 2 and step 3 in the intermediate Z-39 synthesis method, 40-b is taken as a raw material, and is reduced by Pd/C and oxidized by DMP to obtain an intermediate Z-40, MS (ESI) m/z=497 [ M+H ]] ⁺ 。

Synthesis of intermediate Z-41

Step 1, synthesis of intermediate 41-a

To intermediate 40-a (469.02 mg,1.46 mmol) and 37-a (600 mg,1.32 mmol) in DMF (8 mL)/H ₂ K is added into the O (2 mL) mixed solution ₃ PO ₄ (393.32 mg,1.85 mmol), S-phos (65.19 mg, 159.01. Mu. Mol) and Pd (OAc) ₂ (21.21 mg, 92.63. Mu. Mol), the mixture was replaced with nitrogen several times, warmed to 100℃under the protection of nitrogen, and stirred at this temperature for 15 hours. After the completion of the reaction, 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 the crude product was purified by column chromatography over silica gel (PE/ea=2/1) to give 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

To a solution of 41-a (90 mg, 158.25. Mu. Mol) in DCM (2 mL) at room temperature was added TFA (18.04 mg, 158.25. Mu. Mol), the mixture was stirred at room temperature for 2 hours, after the reaction was completed, the reaction mixture was concentrated, the crude product was redissolved in DCM (5 mL), and then TEA (16.01 mg, 158.25. Mu. Mol, 22.07. Mu.L) was added at room temperature, and the mixture was stirred at room temperature for 30 minutes. The reaction was concentrated again and purified by pre-HPLC to give intermediate Z-41 (50 mg, 130.76. Mu. Mol,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-dihydro-isoindol-2-yl) piperidine-2, 6-dione (500 mg,1.92 mmol) in DMF (8 mL) was added 4-bromobutyloxy-tert-butyldimethylsilane (514 mg,1.92 mmol), KHCO ₃ (570 mg,5.76 mmol) and a catalytic amount of KI, the mixture is warmed to 70℃at which temperatureThe reaction was stirred at reflux for 15h. After the completion of the reaction, the reaction mixture was diluted with water, extracted with ethyl acetate, and the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and filtered to give a crude product, which was purified by mHPLC to give intermediate 42-a (160 mg,0.48mmol,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. Mu. Mol) in DCM (10 mL) was added DMP (204.12 mg, 481.42. Mu. Mol) 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 purified by silica gel column (DCM/meoh=10/1) to give 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), cooled to 0℃under nitrogen protection, naH (2.04 g,50.94 mmol) was added, the reaction was continued for 30 minutes, 3-benzyloxybromide (9.34 g,40.76 mmol) was added, the reaction mixture was warmed to 25℃and stirred for 2 hours, after completion of the reaction, saturated aqueous ammonium chloride solution was added to quench, diluted with water, extracted with EA, the combined organic phase was successively washed with water, saturated brine, dried over anhydrous sodium sulfate, and purified by separation with a silica gel column (PE/EA=20/1) after spin-drying to give 43-b (2.5 g,10.15mmol,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), cooled to 0deg.C under nitrogen protection, naH (2.72 g,68.08 mmol) was added, the reaction was continued for 30 min, SEMCl (12.66 g,75.83 mmol) was added, the reaction mixture was warmed to 25deg.C and stirred for 12h, after completion of the reaction, quenched with saturated aqueous ammonium chloride solution, diluted with water, extracted with EA, the combined organic phase was washed successively with water, saturated brine, dried over anhydrous sodium sulfateAfter spin-drying, crude 43-c (28 g,61.76mmol,99.78% yield) was obtained. MS (ESI) m/z=453 [ m+1 ]] ⁺ Directly used 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,45.72mmol,6.37 mL). The reaction solution is N ₂ Stirring at 85deg.C for 12 hr under protection, diluting with water, extracting with EA, washing the separated organic phase with water and saline, drying, and spin-drying. The crude product was purified by column chromatography on silica gel (PE/ea=1/1) to give intermediate 43-d (6.66 g,10.76mmol,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 stirred at room temperature for 12h under the protection of hydrogen after the hydrogen substitution, and after the completion of the reaction, the filtrate was filtered and concentrated and dried to give crude intermediate 43-e (600 mg,1.13mmol,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. Mu. Mol) in DCM (10 mL) was added DMP (238.76 mg, 563.12. Mu. Mol) 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 purified by silica gel column (DCM/meoh=10/1) to give intermediate Z-43 (199mg, 374.96 μmol,99.88% yield). MS (ESI) m/z=531 [ m+h ]] ⁺ 。

Synthesis of intermediate Z-44

Referring to the synthetic route of the intermediate Z-43, the method from step 1 to step 5 uses 4-pentynyl-1-alcohol instead of 5-hexynyl-1-ol-phenylamine as a starting material in step 1, and the rest operations are the same, and the same steps are carried out 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 28-a (50 mg, 78.17. Mu. Mol) of DMF (5 mL)/H ₂ To a solution of 45-a (24.17 mg,78.17 mmol), K was added successively in O (0.5 mL) ₂ CO ₃ (32.36mg,234.5μmol)，Pd(dppf)Cl ₂ (5.71 mg, 7.82. Mu. Mol). The reaction solution is N ₂ Stirring at 90deg.C for 12 hr under protection, diluting with water, extracting with ethyl acetate, washing the separated organic phase with water and saline, drying, and spin drying. The crude product was purified by pre-TLC (100% EA) to give intermediate 45-b (57 mg,76.82mmol,98.28% yield). MS (ESI) m/z=742 [ m+h ]] ⁺ 。

Step 2, synthesis of intermediate Z-45

To a solution of 45-b (32 mg, 43.13. Mu. Mol) in DCM (2 mL) at room temperature was added TFA (18.04 mg, 158.25. Mu. Mol), and the mixture was stirred at room temperature for 2 hours, after completion of the reaction, the reaction solution was concentrated to give crude intermediate Z-45 (22 mg, 43.00. Mu. Mol,99.71% yield) which 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. Mu. Mol) in MeOH (5 mL), the reaction solution was replaced with hydrogen several times, then stirred at room temperature under the protection of hydrogen for 12 hours, after completion of the reaction, filtered, and the filtrate was concentrated and dried to give crude intermediate 46-a (32 mg, 43.01. Mu. Mol,99.73% yield) which was used in the next reaction without purification. MS (ESI) m/z=744 [ m+h ]] ⁺ 。

Step 2, synthesis of intermediate Z-46

To a solution of 46-a (33 mg, 44.35. Mu. Mol) in DCM (2 mL) at room temperature was added TFA (18.04 mg, 158.25. Mu. Mol), and the mixture was stirred at room temperature for 2 hours, after completion of the reaction, the reaction solution was concentrated to give crude intermediate Z-46 (22 mg, 42.83. Mu. Mol,96.57% yield) which 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. Mu. Mol) in DCM (10 mL) was added DMP (267.98 mg, 632.03. Mu. Mol), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was filtered, the filtrate was concentrated, and the crude product was purified by silica gel column (PE/ea=1/2) to give intermediate Z-47 (198mg, 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

48-b (100.07 mg, 838.87. Mu. Mol), DIPEA (280.73 mg,2.17mmol, 378.35. Mu.L) was added to a solution of 48-a (200 mg, 724.06. Mu. Mol) in NMP (10 mL), and the mixture was heated to 90℃under nitrogen to react for 12 hours. After completion of the reaction, water was added, EA was extracted, and the organic phase was concentrated, and the crude product was purified by column chromatography on silica gel (PE/ea=1/4) to give 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. Mu. Mol) in DCM (10 mL) was added DMP (342.50 mg, 807.78. Mu. Mol) and the mixture was stirred at room temperature for 1 hour. After the completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated, and the crude product was purified by silica gel column (PE/ea=1/4) to give 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

To a solution of 2-b (3.0 g,9.28 mmol) in DMF (50 mL), cooled to 0deg.C under nitrogen protection, naH (445.63 mg,11.14 mmol) was added, the reaction was continued for 30 minutes, SEMCl (1.86 g,11.14 mmol) was added, the reaction mixture was warmed to 25deg.C and stirred for 12 hours, after completion of the reaction, saturated aqueous ammonium chloride solution was added to quench, diluted with water, extracted with ethyl acetate, the combined organic phases were successively washed with water, saturated brine, dried over anhydrous sodium sulfate, and the crude product was purified by silica gel column separation (PE/EA=1/1) to give intermediate 49-a (2.97 g,6.55mmol,70.56% yield). MS (ESI) m/z=453 [ m+1 ] ] ⁺ 。

Step 2, synthesis of intermediate 49-c

49-a (0.2 g, 441.11. Mu. Mol), 49-b (127.97 mg, 529.33. Mu. Mol), pd (OAc) ₂ (3.96mg,17.64μmol)，BINAP(21.97mg,35.29μmol)，Cs ₂ CO ₃ (286.72 mg, 882.22. Mu. Mol) was dissolved in a mixed solvent of toluene (10 mL) and t-BuOH (1 mL), and after nitrogen substitution, the mixture was heated to 100℃for reaction for 12 hours. After completion of the reaction, water was added, EA was extracted, and the organic phase was washed successively with water, saturated brine, dried over anhydrous sodium sulfate, and the crude product was purified by separation on a silica gel column (100% EA) to give intermediate 49-c (40 mg, 69.23. Mu. Mol,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. Mu. Mol) in MeOH (5 mL), the reaction solution was replaced with hydrogen several times, then stirred at room temperature for 4 hours under the protection of hydrogen, after completion of the reaction, filtered, and the filtrate was concentrated and dried to give crude intermediate 49-d (36 mg, 73.82. Mu. Mol,94.87% yield) which was 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. Mu. Mol) in DCM (3 mL) was added DMP (46.95 mg, 110.73. Mu. Mol), and the mixture was stirred at room temperature for 1 hour. Reverse-rotationAfter completion of the reaction, the reaction mixture was filtered, the filtrate was concentrated, and the crude product was purified by silica gel column (100% EA) to give intermediate Z-49 (31 mg, 63.83. Mu. Mol,86.47% yield). MS (ESI) m/z=486 [ m+h ] ] ⁺ 。

Synthesis of intermediate Z-50

Step 1, synthesis of 50-a

At zero degrees, 27-g (3.0 g,25.64 mmol) of THF (100 mL)/H ₂ To a solution of O (50 mL), potassium carbonate (10.62 g,76.92 mmol) and Cbz-OSu (12.82 g,51.28 mmol) were added in this order, and after the completion of the reaction, a saturated aqueous sodium hydrogencarbonate solution was added, and the EA extract was separated, the separated organic phase was washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated, and the residue was purified by separation on a silica gel column (50% EA in PE) to give 50-a (5.1 g,20.29mmol,79.13% yield). MS (ESI) m/z=252 [ m+1 ]] ⁺

Step 2, synthesis of 50-b

To a solution of 50-a (5.1 g,20.29 mmol) in DCM (100 mL) was added TEA (6.16 g,60.88mmol,8.49 mL) and MsCl (3.49 g,30.44 mmol) in this order at zero temperature, stirred for 1 hour, after completion of the reaction, saturated aqueous sodium bicarbonate solution was added, the separated organic phase was separated in the following order by water washing, saturated brine washing, dried over anhydrous sodium sulfate and concentrated to give 50-b (6.66 g,20.22mmol,99.63% yield), the crude product was directly reacted with the next step 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 K ₂ CO ₃ (8.40 g,60.84 mmol) was dissolved in DMF (100 mL), the reaction was allowed to warm to 80℃and stirred overnight, after completion of the reaction, cooled to room temperature, EA was added, the mixture was washed 2 times with water, the separated organic phase was concentrated, and the residue was purified by column chromatography over silica gel (50% EA in PE) to give 50-d (3.3 g,6.87mmol,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 a solution of 50-d (3.3 g,6.87 mmol) in MeOH (60 mL), the reaction was replaced with hydrogen several times, then stirred at room temperature for 4h under the protection of hydrogen, after completion of the reaction, filtered, and the filtrate was concentrated and dried to give crude intermediate 50-e (2.16 g,6.24mmol,90.80% yield) which was used directly in the next reaction without purification. MS (ESI) m/z=347 [ m+h ]] ⁺ 。

Step 5, synthesis of 50-g

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

Step 6, synthesis at 50-h

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

Step 7, synthesis of 50-i

To a 50-h (100 mg, 211.43. Mu. Mol) EtOH (3 mL) mixture was added CNBr (111.06 mg,1.06 mmol) and stirred overnight at 60 ℃. After the reaction was completed, saturated aqueous sodium bicarbonate was added, stirred for 10 minutes, the organic phase was separated, washed with saturated aqueous sodium bicarbonate, water, brine in this order, dried over anhydrous sodium sulfate, and the organic phase was concentrated, and the resulting residue was purified by column chromatography on silica gel (DCM/meoh=10/1) to give 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) of THF (50 mL)/H ₂ To 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 completion of the reaction, the reaction mixture was concentrated, diluted with water, and concentrated with 1M HClAdjusting ph=4-5, solid precipitated, filtered, and the filter cake washed with water, dried to give 50-j (1.2 g,2.48mmol,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) were added TEA (1.03 g,10.17mmol,1.42 mL) and TBTU (979.02 mg,3.05 mmol) in sequence, stirred at room temperature for 15 min, after completion of the reaction, diluted with water, separated, the organic phase concentrated and the resulting residue purified by silica gel column separation (DCM/MeOH=10/1) to give intermediate 50-k (1.0 g,2.15mmol,84.44% yield). MS (ESI) m/z=466 [ m+1 ]] ⁺ 。

Step 10, synthesis of intermediate 50-l

To a solution of 50-k (1.0 g,2.15 mmol) in DMF (30 mL) was added DIPEA (832.12 mg,6.44mmol,1.12 mL) and SEMCl (358.42 mg,2.15 mmol) in this order, the reaction mixture was warmed to 60℃and stirred for 12h, after completion of the reaction, the reaction mixture was concentrated, the residue was diluted with water, extracted with ethyl acetate, the combined organic phases were washed successively with water, saturated brine, dried over anhydrous sodium sulfate, and purified by separation on a normal phase silica gel column (100% ethyl acetate) to give 50-l (630 mg,1.06mmol,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. Mu. Mol) in toluene (4 mL)/t-butanol (1 mL) was added 1-t-butoxycarbonylpiperazine (140.58 mg, 754.79. Mu. Mol), BINAP (25.07 mg, 40.26. Mu. Mol), pd (OAc) ₂ (4.51 mg, 20.13. Mu. Mol) and Cs ₂ CO ₃ (327.07 mg,1.01 mmol). The temperature was raised to 120℃under nitrogen 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 give a crude product, which was purified by reverse phase silica gel column separation to give 50-m (370 mg, 495.99. Mu. Mol,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. Mu. Mol) in DCM (10 mL) was added TFA (10 mL) and the mixture was stirred at room temperature for 1 hour. The reaction was then concentrated and the residual TFA was removed by oil pump to give Z-50 (279 mg, 521.71. Mu. Mol)99.79% yield) was 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

NaH (1.43 g,59.42 mmol) was added to a solution of 1-b (16 g,49.51 mmol) in DMF (470 mL) at zero temperature, the mixture was stirred at zero temperature for 1 hour and SEMCl (9.08 g,54.47 mmol) was then added. After the dripping, the mixture reacts for 12 hours at room temperature. After completion of the reaction, diluted with water, extracted with EtOAc, the combined organic phases were washed with water, brine, dried over anhydrous sodium sulfate, and concentrated to give crude 51-a (22 g,48.52mmol,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 for intermediate Z-1, intermediate 51-a is substituted for intermediate 1-b, 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 of intermediate Z-42, in step 1, 4-bromopentyloxy-t-butyldimethylsilane was used in place of 4-bromobutyloxy-t-butyldimethylsilane, and the rest of the procedure was the same, to obtain intermediate Z-52.MS (ESI) m/z=345 [ m+1 ] ] ⁺ 。

Synthesis of intermediate Z-53

Referring to the procedure of steps 11 and 12 in the synthetic route of intermediate Z-50, 1-Boc-4- (piperidin-4-yl) -piperazine was used in place of 1-t-butoxycarbonyl piperazine in step 11, the rest was the sameIntermediate Z-53, MS (ESI) m/z=599 [ M+1 ] can be obtained] ⁺ 。

Synthesis of intermediate Z-54

Referring to the procedure of steps 11 and 12 in the scheme for the synthesis of intermediate Z-50, intermediate 28-a was used in step 11 in place of intermediate 50-l, while tert-butyl 4-piperazin-1-ylpiperidine-1-carboxylate was used in place of 1-tert-butoxycarbonylpiperazine, and the rest of the procedure was the same, to give intermediate Z-54, MS (ESI) m/z=598 [ M+1 ]] ⁺ 。

Synthesis of intermediate Z-55

Referring to the operations of step 5 to step 12 in the route of intermediate Z-50, 2, 4-dichloro-5-nitropyridine was used in place of 2, 6-dichloro-3-nitropyridine in step 5, 4-piperazine-1-ylpiperidine-1-carboxylic acid tert-butyl ester was used in place of 1-tert-butoxycarbonyl piperazine in step 11, and the remaining operations were the same, to obtain intermediate Z-55.MS (ESI) m/z=598 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-56

Referring to the methods of step 11 and step 12 in the synthetic route of intermediate Z-50, in step 11, 1-t-butoxycarbonylpiperazine was replaced with t-butyl 4-piperazin-1-ylpiperidine-1-carboxylate, and the rest was the same, to obtain intermediate Z-56.MS (ESI) m/z=599 [ m+1 ] ] ⁺ 。

Synthesis of intermediate Z-57

Referring to the operations of step 11 and step 12 in the synthetic route for intermediate Z-50, intermediate 50-l was replaced with intermediate 28-a, while 1-t-butoxycarbonyl piperazine was replaced with t-butyl 4- [ (piperazin-1-yl) methyl ] piperidine-1-carboxylate, and the remaining operations were the same, to give 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 for intermediate Z-55, the remaining operations are the same by substituting tert-butyl 4-piperazin-1-ylpiperidine-1-carboxylate with 1-t-butoxycarbonyl piperazine, to obtain intermediate Z-58.MS (ESI) m/z=516 [ m+1]] ⁺ 。

Synthesis of intermediate Z-59

Step 1, synthesis of intermediate 59-a

Referring to the synthesis of intermediate Z-25, intermediate 1-c was replaced with 4-c, and the remaining operating conditions were the same, to afford intermediate 59-a.

Step 2, synthesis of intermediate Z-59

Referring to the synthesis of intermediate Z-26, 59-a was used as starting material instead of Z-25, and intermediate Z-59 was obtained by the same procedure and conditions.

Synthesis of intermediate Z-60

Referring to the procedure of step 1 to step 4 of the intermediate Z-43 synthesis route, in step 1, 7 octynol is used as a starting material to replace 43-a, and the rest of the operation steps are the same, thus obtaining intermediate Z-60, MS (ESI) m/z=559 [ M+1] ] ⁺ 。

Synthesis of intermediate Z-61

Referring to the procedure of example Z-1, synthetic route step 2 to step 4, the 4-pentyn-1-ol was replaced with 10-undecylenol in the initial step, and the remaining procedure was the same, to give intermediate Z-61.MS (ESI) m/z=543 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-62

Referring to step 11 and step 12 of the intermediate Z-50 synthesis route, the 1-t-butoxycarbonylpiperazine was replaced with 3- (piperazin-1-ylmethyl) pyrrolidine-1-carboxylic acid t-butyl ester in the initial step, and the remaining operation steps were the same, to obtain intermediate Z-62.MS (ESI) m/z=599 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-63

Step 1

Z-63-1a (2 g,9.2 mmol) and pyrazole (755 mg,11 mmol) were mixed in acetonitrile (20 ml), acetyl chloride (872 mg,11 mmol) was added dropwise at a temperature of 0℃to 5℃in an ice water bath, then reacted at room temperature for 17 hours, Z-63-1b (1.5 g,9.2 mmol), pyrazole (1.4 g,19.3 mmol) and triphenyl phosphite (3.4 g,11 mmol) were directly added to the reaction solution the next day, then reacted at 85℃for 17 hours, after the completion of the reaction, cooled to room temperature, water was added to the reaction solution, solids were precipitated, the solids were filtered and washed with water 1 time, ethyl acetate 1 time, then the solids were filtered, spun dry, and used directly for the next step.

Step 2

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

Step 3

Z-63-1d(500mg，1.04mmol) and 4-pentyn-1-ol (105 mg,1.25 mmol) were dissolved in DMF and Pd (pph) was added ₃ ) ₂ Cl ₂ (55 mg,0.078 mmol), cuI (30 mg,0.156 mmol) and TEA (315 mg,3.12 mmol) were reacted at 80℃for 17 hours, after the completion of the reaction, water was added, EA was added for extraction and separation, the separated organic phase was washed with water, saturated brine, dried over anhydrous sodium sulfate and concentrated, and the crude product was purified by column chromatography on silica gel (50% EA in PE) to give Z-63-1e (240 mg,0.496mmol,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 reacted at room temperature under hydrogen protection for 17h, after the reaction was completed, the filtrate was filtered and spin-dried to give Z-63-1f (220 mg, 0.457mmol, 90.1% yield) MS (ESI) m/z=488 [ M+1 ]] ⁺

Step 5

Z-63-1f (220 mg, 0.457mmol) was dissolved in methylene chloride, cooled in an ice bath, DMP (229 mg,0.54 mmol) was added, then allowed to react at room temperature for 4h, after completion of the reaction, the solid was filtered off, the filtrate was dried by spinning, and the crude product was purified by column chromatography (PE: EA=2:1) to give compound Z-63 (120 mg,0.246mmol,52% yield), 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 potassium carbonate (1.19 g,8.58 mmol) and tert-butyl bromoacetate (1.1 g,5.71 mmol). After 2h reaction at room temperature, DMF was removed by rotary evaporation, ethyl acetate was dissolved and washed with water, and the organic phase was dried by rotary evaporation. Purification of the crude product on a silica gel column (petroleum ether/ethyl acetate=2/1) afforded intermediate 2- (2, 6-dioxo-piperidin-3-yl) -4-hydroxyacetic acid tert-butyl ester-isoindole-1, 3-dione (2 g,5.1 mmol). This was dissolved in methylene chloride (15 mL), trifluoroacetic acid (10 mL) was added, and the mixture was reacted at room temperature for 1 hour, followed by spin-drying the solvent to give the objective 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 is replaced by 2-amino-5-bromobenzoic acid, and other reagents and operation methods are the same, so as to obtain an intermediate Z-65.

Synthesis of intermediate Z-66

Referring to the procedure of steps 3 to 5 in the synthetic route of intermediate Z-1, 4-pentyn-1-ol is replaced with 3-butyn-1-ol in the initial step, and the remaining operation steps are the same, to obtain intermediate Z-66.MS (ESI) m/z=315 [ m+1 ]] ⁺ 。

Synthesis of intermediate Z-67

Referring to the procedure of intermediate Z-36 synthetic routes step 1 to step 4, the 3-benzyloxy bromopropane was replaced with 2-benzyloxy bromoethane in the initial step, and the remaining 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 of intermediate Z-25, 3-c was used instead of 1-c, and the remaining reagents and procedures were the same, to give intermediate Z-68.MS (ESI) m/z=341 [ m+1 ]] ⁺ 。

EXAMPLE 1 Synthesis of Compound 1

Toward Z-28 (30 mg, 56.75. Mu. Mol)TEA was added to MeOH (2 mL) to pH=9-10, then Z-2 (18.63 mg, 56.75. Mu. Mol) was added, stirred at room temperature for 30 min, then acetic acid was added to adjust pH=4-5. After stirring for 5 minutes, naBH was added in one portion ₃ CN (10.73 mg, 170.25. Mu. Mol) and the reaction mixture was allowed to stand overnight at room temperature. After completion of the reaction, the reaction solution was filtered, and the filtrate was separated and purified by pre.hplc to give example 1 (1.42 mg,1.59 μmol,2.80% yield, 94% purity). MS (ESI) m/z=837 [ m+1 ] ] ⁺ ， ¹ H NMR(400MHz,DMSO-d ₆ )δ8.43(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 Synthesis of Compound 2

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Z-28 (46.60 mg, 88.15. Mu. Mol) and Z-25 (30 mg, 88.15. Mu. Mol) were miscible in DMF (5 mL) and DIPEA (56.96 mg, 440.75. Mu. Mol, 76.77. Mu. L) and HATU (40.22 mg, 105.78. Mu. Mol) were then added sequentially. The reaction mixture was stirred at room temperature for 4 hours, after completion of the reaction, 1mL of MeOH was added, and the crude product obtained by concentrating the reaction mixture was separated and purified by pre.HPLC to give example 2 (1.32 mg, 1.37. Mu. Mol,1.55% yield). MS (ESI) m/z=851 [ m+1 ]] ⁺ ， ¹ H NMR(400MHz,CD ₃ OD)δ8.86(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＝13.3,5.2Hz,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 Synthesis of Compound 3

Referring to the synthesis of example 1, intermediate compounds Z-28 and Z-16 were used as starting materials for reductive amination to give example 3.MS (ESI) m/z=851 [ m+1 ]] ⁺ ， ¹ H NMR(400MHz,DMSO-d ₆ )δ8.43(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 Synthesis of Compound 4

With reference to the synthetic method of example 1, intermediate Z-14 and intermediate Z-28 were used as starting materials, and reductive amination was performed to give 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 Synthesis of Compound 5

With reference to the synthesis of example 2, example 5 was obtained by condensation using intermediates Z-26 and Z-28 as starting materials and HATU as condensing agent. 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 Synthesis of Compound 6

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Referring to the synthesis of example 1, using Z-1 and Z-28 as starting materials, reductive amination gave 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).

Examples 7 to 27 Synthesis of Compounds 7 to 27

Reference is made to the reductive amination process of example 1, with the corresponding SM-aldehyde in the table replacing Z-2 in step. The following table examples 7 to 27 were obtained by substituting SM-amine for Z-28 in the step and the rest of the procedure was the same:

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EXAMPLE 27 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 (4478 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) and the mixture was purged several times with nitrogen, then with N ₂ The temperature was raised to 90℃under the protection of (C) and the reaction was stirred at the temperature again for 12h. After the completion of the reaction, the reaction mixture was concentrated, the crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and the concentrated crude product was purified by column chromatography on silica gel (100% ea) to give 27-1 (190 mg,0.56mmol,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 (470 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, and the filtrate was concentrated, and the obtained crude product was purified by silica gel column (DCM/meoh=10/1) to give 27-2 (134 mg,0.39mmol,71% yield). MS (ESI) m/z=340 [ m+h ] ] ⁺ 。

Step 3 Synthesis of Compound 27

To intermediate Z-28 (70 mg,0.13 mmol) in DCM (7 mL) was added 27-2 (58.4 mg,0.17 mmol) and a catalytic amount of acetic acid, and the mixture was stirred at room temperature for 1h, followed by NaBH (OAc) ₃ (84 mg,0.397 mmol) and the reaction was stirred at room temperature for 4 hours. After the completion of the reaction, the reaction mixture 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 give compound 27 (45 mg,0.05mmol, yield 40%). MS (ESI) m/z=852 [ m+h ]] ⁺ 。

EXAMPLE 28 Synthesis of Compound 28

Referring to the procedure of step 1 to step 3 in the synthesis of example 27, intermediate Z-33 was used in place of intermediate Z-35 in step 1, and the remaining procedure was unchanged, to give compound 28.MS (ESI) m/z=852 [ m+h ]] ⁺ 。

EXAMPLE 29 Synthesis of Compound 29

Referring to the procedure of step 1 to step 3 in the synthesis of example 27, intermediate Z-34 was used in place of intermediate Z-35 in step 1, and the remaining procedure was unchanged, to give compound 29.MS (ESI) m/z=852 [ m+h ]] ⁺ 。

Examples 30 to 38, synthesis of Compounds 30 to 38

Reference is made to the procedure of step 3 reductive amination in the synthesis of example 27, with substitution of the corresponding SM-aldehyde in the table for 27-2 in step. The following table examples 30 to 38 were obtained by substituting SM-amine for Z-30 in the step and the rest of the procedure was the same:

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EXAMPLE 39 Synthesis of Compound 39

Step 1, synthesis of intermediate 39-1

To a mixture 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 under the protection of hydrogen for 16 hours. After completion of the reaction, pd/C was removed by filtration, and the filtrate was concentrated to give 39-1 (181 mg,0.52mmol,78% yield, crude).

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 completion of the reaction, the mixture was filtered, the filtrate was concentrated to dryness, and the crude product was diluted with ethyl acetate to precipitate a brown solid, and the solid was collected by filtration and purified by reverse phase C18 column to give intermediate compound 39-2 (116 mg,0.34mmol,65% yield). MS (ESI) m/z=344 [ m+1 ]] ⁺ 。

Step 3 Synthesis of Compound 39

Referring to example 27, step 3, intermediate 39-2 was used in place of 27-2, and the remaining procedure was identical (i.e., in NaBH (AcO) ₃ To reagent, DCM is added with a catalytic amountAcetic acid as solvent, reductive amination) to give compound 39.MS (ESI) m/z=856 [ m+1 ]] ⁺ 。

EXAMPLE 40 Synthesis of Compound 40

Referring to the procedure of step 1 to step 3 in the synthesis of example 39, intermediate 40-2 was used in place of intermediate 39-2 in step 1, and the remaining procedure was unchanged, to give compound 28.MS (ESI) m/z=856 [ m+h ] ] ⁺ 。

EXAMPLE 41 Synthesis of Compound 41

Referring to the procedure of step 1 to step 3 in the synthesis of example 39, intermediate 29-1 was used in place of intermediate 27-1 in step 1, and the remaining procedure was unchanged, to give compound 41.MS (ESI) m/z=856 [ m+h ]] ⁺ 。

Examples 42 to 50, synthesis of Compounds 42 to 50

Reference is made to the procedure of step 3 reductive amination in the synthesis of example 27, with substitution of the corresponding SM-aldehyde in the table for 27-2 in step. The following table examples 42 to 50 were obtained by substituting SM-amine for Z-30 in the step and the rest of the procedure was the same:

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example 51 Synthesis of Compound 51 into Z-28

Step 1, synthesis of intermediate 51-1

TFA was added dropwise to a mixture of Z-28 (42.58 mg, 80.54. Mu. Mol) and Z-40 (42.58 mg, 80.54. Mu. Mol) in DCM (2 mL), the pH was adjusted to 5 to 6, and after stirring for 30min, naBH (OAc) was added ₃ (51.21 mg, 241.63. Mu. Mol), the mixture was stirred at room temperature for 15 hours. After completion of the reaction, the reaction mixture was diluted with water, extracted with DCM, the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and the concentrated crude product was isolated and purified by pre-HPLC to give 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. Mu. Mol) in DCM (2 mL) at room temperature was added TFA (1.36 mg, 11.89. Mu. Mol,2 mL) and the mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction mixture was concentrated, and the crude product was redissolved in DCM (2 mL), TEA (725.50 mg,7.17mmol,1 mL) was added at room temperature and the reaction was stirred for 30 min. The mixture was concentrated and the crude product was isolated and purified by pre-HPLC to give 51 (1.14 mg,1.14umol,9.56% yield, 99% purity, TF) as a yellow solid. MS (ESI) m/z=879 [ m+h ] ] ⁺ . Nuclear magnetic data: 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 Synthesis of Compound 52

Synthesis of reference Compound 51 step 1And step 2, using an intermediate Z-30 to replace Z-28 in the step 1 as a starting material, and obtaining the compound 52 by the same operation method. MS (ESI) m/z=879 [ m+h ]] ⁺ . Nuclear magnetic data: 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 Synthesis of Compound 53

Referring to the synthesis of step 1 of example 51, reductive amination using Z-28 and Z-36 as starting materials gives compound 53, MS (ESI) m/z=879 [ M+H ]] ⁺ . Nuclear magnetic data: 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).

examples 54 to 62, synthesis of Compounds 54 to 62

With reference to the procedure of step 1 of example 51, the compounds of the corresponding examples in the tables are obtained by reductive amination starting from the SM-aldehyde and SM-amine in the tables below.

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EXAMPLE 63 Synthesis of Compound 63

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

After the completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated and purified by pre-HPLC to give compound 63 (9.38 mg,0.009mmol,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).

Examples 64 to 74, synthesis of Compounds 64 to 74

With reference to the method of example 63, the compounds of the corresponding examples in the tables can be obtained by condensation under the same conditions starting from SM-amine and SM-acid in the tables below.

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EXAMPLE 75 Synthesis of Compound 75

To a solution of Z-30 (50 mg, 94.58. Mu. Mol) in DCM (4 mL) was added Z-42 (31.24 mg, 94.58. Mu. Mol), and the reaction mixture was stirred at room temperature for 30min, followed by dropwise addition of AcOH to adjust the pH of the reaction solution to 3 to 4. NaBH (AcO) is added again ₃ (60.14 mg, 283.74. Mu. Mol) and the mixture was stirred at room temperature for 5 hours. After completion of the reaction, the reaction mixture was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and the crude product obtained after concentration was separated and purified by pre.HPLC to give compound 75 (33.8 mg, 35.32. Mu. Mol,37.34% yield, TF), MS (ESI) m/z=843 [ M+H ]]+. Nuclear magnetic data: 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).

examples 76 to 78, synthesis of Compounds 76 to Compounds

With reference to the method of example 75, the compounds of the corresponding examples in the tables were obtained by reductive amination under the same conditions starting from SM-amine and Z-42 in the tables below.

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Examples 79 to 81, synthesis of Compounds 79 to 81

Referring to the method of example 75, the compounds of the corresponding examples in the following table were obtained by reductive amination under the same conditions using intermediate Z-28 as the starting material and the corresponding structural aldehyde in the following table, respectively.

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EXAMPLE 82 Synthesis of Compound 82

Referring to the procedure of example 63, starting from intermediate Z-28 and SM-acid in the following Table, example 82 was obtained by condensation under the same conditions.

Examples 83 to 85, synthesis of Compounds 83 to 85

Referring to the method of example 75, the compounds of the corresponding examples in the following table were obtained by reductive amination under the same conditions using intermediate Z-30 as the starting material and the corresponding structural aldehyde in the following table, respectively.

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Examples 86 to 91, synthesis of Compounds 86 to 91

Referring to the method of example 75, the compounds of the corresponding examples in the following table were obtained by reductive amination under the same conditions using intermediate Z-31 as the starting material and the corresponding structural aldehyde in the following table, respectively.

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EXAMPLE 91 Synthesis of Compound 91

Referring to the procedure of example 63, starting from intermediate Z-31 and SM-acid in the following Table, example compound 91 was obtained by the same condensation reaction.

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Example 92, example 93, synthesis of Compound 92, compound 93

Referring to the method of example 75, the compounds of the corresponding examples in the following table were obtained by reductive amination under the same conditions using intermediate Z-32 as the starting material and the corresponding structural aldehyde in the following table, respectively.

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EXAMPLE 94 Synthesis of Compound 94

Step 1, synthesis of intermediate 94-a

To Z-31 (36 mg, 67.83. Mu. Mol) in DCM (2 mL) was added TEA to pH=9-10, then Z-43 (41.89 mg, 81.40. Mu. Mol) was added, stirred at room temperature for 30 min, then acetic acid was added to adjust pH=4-5. After stirring for 5 minutes, naBH (OAc) was added in one portion ₃ (43.13 mg, 203.49. Mu. Mol) and the reaction mixture was allowed to stand overnight at room temperature. After completion of the reaction, water was added, DCM was added to extract, the organic phase was washed with water, brine, dried, and concentrated, and the crude product was purified by Pre-TLC (DCM/meoh=10/1) to give intermediate 94-a (69 mg,67.03 μmol,98.82% yield). MS (ESI) m/z=1029 [ m+1 ]] ⁺ 。

Step 2 Synthesis of Compound 94

TFA (18.04 mg, 158.25. Mu. Mol) was added to a solution of 94-a (70 mg, 68.00. Mu. Mol) in DCM (2 mL) at room temperature, the mixture was stirred at room temperature for 2 hours, after the reaction was completed, the reaction mixture was concentrated, and the crude product was purified by pre-HPLC to give 94; (30.88 mg, 28.96. Mu. Mol,42.58% yield, 95% purity, TF) yellow solid. MS (ESI) m/z=899 [ m+h ]] ⁺ . Nuclear magnetic data: 1H NMR (400 MHz, methanol-d 4) delta 8.86 (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=2.1 Hz, 1H), 7.05 (d, J=8.7 Hz, 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=5.6 hz, 2H), 3.46 (t, j=6.3 hz, 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).

Examples 95 to 101, synthesis of Compounds 95 to 101

By the method of step 1 to step 2 of example 94, the compounds of the corresponding examples in the following tables were obtained by reductive amination using SM-aldehyde and SM-amine as starting materials.

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

Test example 1 EGFR kinase Activity assay

EGFR kinase activity is characterized by detecting the level of phosphorylation of the substrate, and the strength of the FRET signal between the phosphorylated antibody and the biotin-labeled polypeptide substrate is proportional to the enzyme activity. The reagents required for detection are contained in the protein

KinEASE ^TM In the kit (cisbaio), the specific procedures are as follows: 125pM EGFR (T790M/C797S/L858R) kinase protein, 300nM TK Substrate-biotin, 0.8. Mu.M ATP was added to 384 well plates, the total reaction volume was 10. Mu.L, and incubated at room temperature for 40 minutes after mixing; then, 37.5nM strepavidin-XL 665 and 5. Mu.L/well TK Antibody-Cryptate were added, the final volume of the reaction was 20. Mu.L, and after mixing, the reaction was incubated at room temperature for 60 minutes, and fluorescence intensities at excitation wavelengths of 320nM and emission wavelengths of 620nM and 665nM were detected by an enzyme-labeled instrument, and FRET signal intensities (665 nM fluorescence intensity/620 nM fluorescence intensity: 10000) were calculated. When the compound is detected to inhibit kinase activity, the compound is pre-incubated with kinase protein for 15-60 min at room temperature, and then substrate and ATP are added for reaction. The activity data are shown in the following table: / >

Test example 2 EGFR-PROTAC-CRBN/DDB1 ternary complex formation assay

Serial concentrations of the compound of example and EGFR (T790M/C797S/L858R) recombinant protein and CRBN/DDB1 recombinant protein complex were added to 384 well plates, the reaction buffer was 1×pbs pH 7.4, incubated at room temperature for 120 min, 5nM Anti GST-Eu cryptate and 6.67nM, ant 6his-XL665 were added, mixed, incubated at room temperature for 60min, and fluorescence intensities at excitation wavelengths 320nM at 620nM and 665nM were detected with an enzyme-marker instrument, and FRET signal intensities (665 nM fluorescence intensity/620 nM fluorescence intensity 10000) were calculated. The results of the experiment are shown in figure 1, and the compounds of the invention can form ternary complexes.

Test example 3, cell level EGFR degradation assay

After the 96-hole transparent bottom blackboard is coated by collagen and fibronectin, ba/F3-EGFR (T790M/C797S/L858R) over-expression cells (2 x 10≡4 cells per hole) are inoculated and treated at 37 ℃ with 5% CO ₂ Culturing overnight in an incubator. The next day, the medium in the cells was removed, 150. Mu.L of fresh medium was added, then 50. Mu.L of medium containing 4 times the final concentration of the compound of example was added, and placed in an incubator at 37℃and 5% CO ₂ The culture was continued for 16 hours under the conditions. After that, the medium in the cells was removed, and the cells were fixed with 4% paraformaldehyde, 150. Mu.L per well, and incubated at room temperature for 15min. Cells were permeabilized after cell fixation using 0.1% (v/v) Triton X-100, 150. Mu.L per well, and incubated at room temperature for 15min. Cells were blocked with 1% blocking reagent (Roche, cat# 11096176001), 100. Mu.L per well, and incubated at room temperature for 60min. 50. Mu.L of EGFR antibody (CST, cat# 4267S, dilution ratio 1:250) diluted with 1% (w/v) blocking reagent was added to each well after blocking and incubated for 2 hours at room temperature. EGFR antibody was removed from wells, cells were washed with 0.05% (v/v) PBST, 200. Mu.L per well, and repeated 4 times. Then, 1% (w/v) of a blocking reagent diluted goat anti-rabbit fluorescent secondary antibody (Invitrogen, cat# A32731) and DAPI dye (Thermofish, cat# 62247) were added (fluorescent secondary antibody dilution ratio 1:250, DAPI dilution ratio 1:2000), 50 μl per well, and incubated at room temperature for 2 hours in the absence of light at 300rpm on a shaker. The antibody in the wells was then removed and the cells were washed with 0.05% (v/v) PBST, 200 μl per well, and repeated 4 times (the wash in the last well was retained). Data were acquired using a high content imaging analysis system (GE, in Cell Analyzer, model: 2200), with 9 fields scanned per well, FITC channel exposure time set to 0.5s, and DAPI channel exposure time set to 0.05s. The data were analyzed using In cell analyzer workstation software to obtain the EGFR fluorescent signal values in the cells and the background values around the cells. Subsequent processing of the obtained data using Excel, first The normalized EGFR fluorescent signal value is obtained by subtracting the background value around the cell from the EGFR fluorescent signal value in the cell. Second, relative levels of EGFR after compound treatment (100% DMSO treatment) relative to DMSO control were calculated, analyzed using GraphPad Prism software, and DCs were obtained ₅₀ Values and fitting curves.

Experimental results show that the compound can inhibit and degrade EGFR protein, and can be a drug for targeting EGFR protein degradation.

Claims

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

R ¹ selected from methyl; r is R ² Selected from methyl; x is X ¹ Selected from O; x is X ² Selected from CH ₂ ；

Ring a is selected from pyrazoles; wherein pyrazole may be further substituted by one R ^A Substitution; r is R ^A Each independently selected from hydrogen, methyl;

y is selected from N;

R ⁴ 、R ⁵ each independently selected from hydrogen;

Z ¹ 、Z ² 、Z ³ 、Z ⁴ each independently selected from CH or N;

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

each R ^B Each independently selected from hydrogen, halogen, cyano, nitro, =o, =s, methyl;

l is selected from the following groups:

-(C _0～2 alkylene) - (5-to-10 degree6 membered heterocycloalkyl) - (C _0～10 Alkylene) - (CH≡CH) -,

-(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) - (C _0～10 Alkylene radicals)

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

-(C _0～2 alkylene) - (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 heterogeniesCycloalkyl) -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) -.

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

the B ring is selected from

3. The bifunctional compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: the compound of formula I is shown as formula IIa:

Y is selected from N;

R ⁴ 、R ⁵ each independently selected from hydrogen;

Z ¹ 、Z ² 、Z ³ 、Z ⁴ each independently selected from CH or N;

l is selected from the following groups:

-(C _0～2 alkylene) - (5-to-10 degree6 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 (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) -.

4. The bifunctional compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: the compound of the formula I is shown as a formula IIb:

Y is selected from N;

R ⁴ 、R ⁵ each independently selected from hydrogen;

Z ¹ 、Z ² 、Z ³ 、Z ⁴ each independently selected from CH or N;

l is selected from the following groups:

-(C _0～2 Alkylene) - (5-6 membered heterocyclic ring Alkyl) - (5-6 membered heterocycloalkyl) - (C _0～10 Alkylene) - (5-6 membered aromatic heterocycle) -, - (C _0～2 Alkylene) - (5-6 membered heterocycloalkyl) - (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) -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) -;

R ^B selected from hydrogen, halogen, cyano, nitro, =o, =s, methyl.

5. The bifunctional compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: the compound of the formula I is shown as a formula IIc:

Y is selected from N;

R ⁴ 、R ⁵ each independently selected from hydrogen;

Z ¹ 、Z ² 、Z ³ 、Z ⁴ each independently selected from CH or N;

w is selected from C (O), CH ₂ 、NH；

L is selected from the following groups:

-(C _0～2 alkylene) - (5-to-10 degree6 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 (O) - (C) _0～10 Alkylene) - (CH≡CH) -, - (C _0～2 Alkylene) - (5-6 membered heterocycloalkyl) -C (O) - (C) _0～10 Alkylene group) -,

-(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) -.

6. The bifunctional compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein: the method is characterized in that:

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 group) -,

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

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

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

-(C _0～1 alkylene) - (piperidine) - (C _0～10 Alkylene group) -,

-(C _0～1 Alkylene) - (piperazine) -C (O) - (C _0～10 Alkylene radicals)

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

-(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- (3-4 membered cycloalkane) -.

7. The bifunctional compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein: the compound is specifically as follows:

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8. use of a bifunctional compound of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a tumor or cancer.

9. A composition comprising the bifunctional compound of any one of claims 1-7, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.