CN114057770A

CN114057770A - Bifunctional compounds targeting EGFR protein degradation

Info

Publication number: CN114057770A
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: 2022-02-18
Anticipated expiration: 2041-08-06
Also published as: CN114057770B

Abstract

The invention provides a novel bifunctional compound, or a stereoisomer and a pharmaceutically acceptable salt thereof, and application thereof in preparing medicaments. The bifunctional compound is a protein targeting union, can selectively promote ubiquitination and degradation of EGFR protein, 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 for targeted EGFR protein degradation and application thereof in preparation of medicines.

Background

Egfr (epidemal Growth Factor receptor) is a member of the erbB receptor family of transmembrane protein tyrosine kinases. EGFR can 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 lead to phosphorylation of key tyrosine residues in EGFR cells, thereby activating multiple downstream signaling pathways in the cells. These intracellular signaling pathways play important roles in cell proliferation, survival, and resistance to apoptosis. Dysregulation of the EGFR signaling pathway, including increased expression of ligands and receptors, EGFR gene amplification and mutation, can promote cellular transformation to malignancy, 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 anti-cancer drugs.

The Ubiquitin protein (Ubiquitin) itself consists of 76 amino acid residues with a molecular weight of about 8.5kDa, and is named "Ubiquitin" because it is widely found in eukaryotic organisms. Ubiquitination (Ubiquitination) refers to a process in which ubiquitin molecules classify proteins in cells under the action of a series of special enzymes, target protein molecules are selected from the proteins, and the target proteins are subjected to specific modification to form target protein polyubiquitin chains. These specific enzymes include ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), ubiquitin conjugating enzyme (E3), and the like. Ubiquitination is closely related to the onset of diseases such as tumor, cardiovascular disease, autoimmunity, etc. Degradation of proteins by the ubiquitin-proteasome system is an important pathway for the selective degradation of proteins and can be simply divided into three steps: firstly, a target protein is added with a ubiquitin tag by a ligase, then a plurality of ubiquitin tags are obtained after a plurality of rounds of ubiquitination, and the protein after polyubiquitination is identified by a protease body and is 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 two ends are connected through a proper connecting chain, so that the degradation of the target protein can be effectively promoted.

Most of the marketed EGFR inhibitor drugs develop resistance to small molecule inhibitors after treatment. The drug resistance mechanism comprises EGFR secondary mutation, bypass activation and the like. At present, no medicine for degrading EGFR protein by constructing a bifunctional compound is on the market. The development of the bifunctional compound capable of ubiquitinating and degrading EGFR protein can become a new method for treating tumors or cancers, and has wide application prospect.

Disclosure of Invention

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

wherein,

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

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

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

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

y is selected from N or CR^Y1；

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

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

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

R⁴、R ⁵0, 1,2 or 3 are respectively independent;

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

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

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

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

l is- (L)^A)_q-；

q is an integer of 1 or more and 100 or less;

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

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

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

Further, the air conditioner is provided with a fan,

ring B is selected from

Or

Further, the air conditioner is provided with a fan,

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

Wherein L is^A1One end is connected with

Is connected with the other end of L^A2Connecting;

L^A7one end and L^A6Is connected with the other end of the other end

Connecting;

L^A1is selected from- (C)_0～2Alkylene) -;

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

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

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

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

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

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

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

Further, in the present invention,

L^A3is selected from

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

Or none.

Further, the air conditioner is provided with a fan,

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

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

R⁴、R⁵each independently selected from hydrogen.

Further, the compound of formula I is represented by formula IIa:

wherein,

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

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

y is selected from N or CR^Y1；

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

R⁴、R ⁵0, 1,2 or 3 are respectively independent;

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

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

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

Further, the compound of formula I is represented by formula IIb:

wherein,

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

y is selected from N or CR^Y1；

R⁴、R ⁵0, 1,2 or 3 are respectively independent;

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

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

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

Further, the compound of formula I is represented by formula IIc:

wherein,

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

y is selected from N or CR^Y1；

R⁴、R ⁵0, 1,2 or 3 are respectively independent;

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

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

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

In a still further particular embodiment, the first and second,

l is selected from the following groups:

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Still more particularly, it is contemplated that the first,

R⁴、R⁵each independently selected from hydrogen;

w is selected from C (O), CH₂、NH；

R^BIs selected from methyl;

l is selected from the following linking groups:

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

end 1 in the above structure and

connecting; end 2 in the above structure and

are connected.

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

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

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

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

Definitions of terms used in connection with the present invention: the initial definitions provided herein for a group or term apply to that group or term throughout the specification unless otherwise indicated; for terms not specifically defined herein, the meanings that would be given to them by a person skilled in the art are to be given in light of the disclosure and the context.

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

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

Minimum and maximum values of the carbon atom content in the hydrocarbon groupOver-prefix representation, e.g. prefix C_a～bAlkyl means any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C_1～4The alkyl group means an alkyl group having 1 to 4 carbon atoms.

"alkyl" refers to a saturated hydrocarbon chain having the indicated number of member atoms. E.g. C₁～₆Alkyl 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 C₁～₆An alkoxy group.

"alkylene" as used herein refers to a divalent saturated aliphatic hydrocarbon group having from 1 to 10 carbon atoms and in some embodiments from 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～C₆) Alkylene "is intended to include methylene, ethylene, propylene, 2-methylpropylene, dimethylethylene, pentylene, and the like. Thus, the term "propylene" can be exemplified by the following structure:

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

or

Furthermore, the term "(C)₁～C₆) Alkylene "is intended to include such branched alkyl groups, such as cyclopropylAn oxymethylene group, which can be exemplified by the following structure:

"cycloalkyl", "cycloalkane" as used herein refers to saturated or partially saturated cyclic groups 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 including polybicycloalkyl ring systems are bicyclohexyl, bicyclopentyl, bicyclooctyl and the like. Two such bicycloalkyl polycyclic structures are exemplified and named below:

dicyclohexyl and

a dicyclohexyl group. Adamantyl includes, but is not limited to, the following structures:

"alkenyl" refers to a straight or branched chain hydrocarbyl 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 vinyl 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.

The "alkenylene group" as used herein refers to a hydrocarbon chain having 2 to 10 carbon atoms, at least one double bond and two unsaturated chemical valencies. For example, (C)₃-C₆) Alkenylene radicals include>C＝CH-CH₂-、-CH-CH＝CH-CH₂-and the like.

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

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

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

"haloalkyl", "halo-substituted alkyl" means that a hydrogen atom in an alkyl group may be substituted with one or more halo atoms. E.g. C_1～4The haloalkyl group means an alkyl group having 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 atom OR the nitrogen atom by a single bond.

The "-C (O) R", "-S (O)" mentioned in the present invention₂The oxygen atom in R' or the like is bonded to a carbon atom or a sulfur atom with a double bond.

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

As used herein, "heterocycle", "heterocycloalkyl", "heterocycloalkane" 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. Generally denotes a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of a plurality of ring atoms, preferably a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of 3 to 9 ring atoms, comprising 1,2 or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Bicyclic means consisting of two rings 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, piperazinyl, thiazolyl, morpholinyl, piperazinyl, morpholinyl, piperazinyl, or the like,

Thiomorpholinyl, 1-dioxo-thiomorpholin-4-yl, azepinyl, diazepanyl, homopiperazinyl or oxazepinyl. An example of a bicyclic saturated heterocycloalkyl 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 radical,

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

The "aromatic ring", "aryl" as used herein refers to an aromatic hydrocarbon group having a plurality of carbon atoms. The aryl group is typically a monocyclic, bicyclic or tricyclic aryl group having 5 to 20 carbon atoms. Further, 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.

The term "aromatic heterocycle", "aromatic heterocyclic group" as used herein means an aromatic unsaturated ring containing at least one hetero atom; wherein the hetero atom means a nitrogen atom, an oxygen atom, a sulfur atom, etc. Aromatic monocyclic or bicyclic hydrocarbons which typically contain multiple ring atoms, wherein one or more ring atoms are selected from the group consisting of the heteroatoms of O, N, S. Preferably there are one to three heteroatoms. Heterocyclic aryl 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" includes 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 a pharmaceutical dosage form and physiologically compatible with the recipient.

The terms "salt" and "pharmaceutically acceptable salt" refer to acid and/or base salts of the above compounds or stereoisomers thereof, with inorganic and/or organic acids and bases, as well as zwitterionic (inner) salts, and also quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. The compound or a stereoisomer thereof may be obtained by appropriately (e.g., equivalently) mixing the above compound or a stereoisomer thereof with a predetermined amount of an acid or a base. These salts may form precipitates in the solution which are collected by filtration, or they may be recovered after evaporation of the solvent, or they may be prepared by reaction in an aqueous medium followed by lyophilization. The salt in the invention can be hydrochloride, sulfate, citrate, benzene sulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

Drawings

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

Detailed Description

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Known starting materials for the present invention can be synthesized by or according to methods known in the art, or can be purchased from companies such as Enduragi chemistry, Chengdulong chemistry, Shaoshi chemistry technology, and Bailingwei technology.

The reagents described in the examples are abbreviated as follows: AIBN: azodiisobutyronitrile; NBS: n-bromosuccinimide; DMP: DESS-MARTIN peeriodinane DESS-MARTIN oxidizer; DIPEA: n, N-diisopropylethylamine; TEMPO: 2,2,6, 6-tetramethylpiperidine oxide; MTBE: methyl tert-butyl ether; NaHMDS: sodium bis (trimethylsilyl) amide; the DIAD: diisopropyl azodicarboxylate; DPPA: diphenyl phosphorazidate; TBTU: 2- (1H-benzotriazol L-1-yl) -1,1,3, 3-tetramethyluronium tetrafluoroborate; TEA: triethylamine; SEMCl: 2- (trimethylsilyl) ethoxymethyl chloride; BINAP: 1,1 '-binaphthyl-2, 2' -bis-diphenylphosphine; X-Phos: 2-dicyclohexylphosphonium-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.

In the examples, the reaction was carried out under a nitrogen atmosphere without specific mention. In the examples, the solution means an aqueous solution unless otherwise specified. In the examples, the reaction temperature is room temperature, unless otherwise specified. The room temperature is the most suitable reaction temperature and is 20-30 ℃. In the examples, M is mole per liter, unless otherwise specified.

The structure of the compounds was determined by Nuclear Magnetic Resonance (NMR) and Mass Spectrometry (MS). NMR shifts (. delta.) are given in units of 10-6 (ppm). NMR was measured using (Bruker AvanceIII 400 and Bruker Avance 300) nuclear magnetic instrument in deuterated dimethyl sulfoxide (DMSO-d)₆) Deuterated chloroform (CDCl)₃) Deuterated methanol (CD)₃OD), internal standard Tetramethylsilane (TMS). LC-MS was measured using Shimadzu LC-MS 2020 (ESI). HPLC was performed using Shimadzu high pressure liquid chromatograph (Shimadzu LC-20A). MPLC (Medium pressure preparative chromatography) Gilson GX-281 reverse phase preparative chromatography was used. The thin layer chromatography silica gel plate is a tobacco yellow sea HSGF254 or Qingdao GF254 silica gel plate, and the specification of the thin layer chromatography separation and purification product is 0.4 mm-0.5 mm. The column chromatography generally uses 200-300 mesh silica gel of the Tibet Huanghai silica gel as a carrier.

Synthesis of intermediate Z-1

Synthesis of step 1, 1-a

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

Synthesis of Steps 2, 1-b

To a solution of 1-a (42g,136.38mmol) and 3-aminopiperidine-2, 6-dione hydrochloride (17.47g,136.38mmol) in acetonitrile (210mL) was added DIPEA (21.15g,163.65mmol,28.51mL), the reaction 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 (35.6g,110.17mmol, 80.78% yield). MS (ESI) M/z 324[ M +1]]⁺。

Synthesis of Steps 3, 1-c

To a solution of 1-b (3g,9.28mmol) in DMF (30mL) was added 4-pentyn-1-ol (1.56g,13.93mmol), cuprous iodide (265.22mg,1.39mmol), Pd (Ph) in that order₃P)₂Cl₂(488.10mg, 696.29. mu. mol) and DIPEA (2.82g,27.85mmol,3.88 mL). Reaction solution in N₂Stirring at 80 deg.C for 18 hr under protection, diluting with water, extracting with ethyl acetate, washing the separated organic phase with water and brine, drying, and spin-drying. Ethyl acetate was added to the crude product to precipitate a brown solid, which was then filtered to collect a filter cake and obtain the objective compound 1-c (1.6g,4.51mmol, 48.65% yield). MS (ESI) M/z 355[ M +1]]⁺。

Synthesis of Steps 4, 1-d

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

Step 5, synthesis of intermediate Z-1

To a mixed solution of Compound 1-d (300mg, 881.39. mu. mol) in DCM (3mL) and DMF (3mL) was added dessimutant reagent (560.75mg,1.32 mmol). The suspension was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was filtered, the filtrate was concentrated and dried, the crude product was diluted with ethyl acetate to precipitate a brown solid, which was collected by filtration and separated and purified by reverse phase C18 column to obtain intermediate compound Z-1(200mg, 591.09. mu. mol, 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 to obtain Z-2. MS (ESI) M/z 339[ M +1]]⁺。

Synthesis of intermediate Z-3

Following steps 1 through 5 of the synthetic method for intermediate Z-1, methyl 2-methyl-3-bromobenzoate in step 1 was replaced with methyl 2-methyl-5-bromobenzoate, and the remaining conditions were unchanged to give Z-3. MS (ESI) M/z 339[ M +1]]⁺。

Synthesis of intermediates Z-4 to Z-12

According to the steps 1 to 5 in the synthesis method of the intermediate Z-1, respectively replacing 2-methyl-3-bromobenzoate methyl in the step 1 with SM-Br in the table, replacing 4-pentyne-1-ol in the step 3 with corresponding SM-ene in the table, and keeping the other conditions unchanged to obtain the corresponding structure of Z-4 matched with Z-12 in the table.

Synthesis of intermediate Z-13

To a mixed solution of intermediate compound 1-c (500mg,1.53mmol) in DMF (4mL)/DCM (40mL) was added DMP (974.75mg,2.30mmol) at zero degrees, the reaction mixture was slowly warmed to room temperature and stirred for 3 hours, after completion of the reaction, 1mL methanol was added, the reaction mixture was spun off, and the residue was isolated and purified by m-HPLC to give intermediate compound Z-13(133mg, 410.08. mu. mol, 26.77% yield). MS (ESI) M/z 325[ M +1]]⁺。

Synthesis of intermediate Z-14

Referring to the synthesis method of compound Z-13, starting material 1-c was replaced with 2-c, and the remaining method was unchanged to give Z-14. MS (ESI) M/z 325[ M +1]]⁺。

Synthesis of intermediate Z-15

Referring to the synthesis method of compound Z-13, starting material 1-c was replaced with 3-c, and the remaining method was unchanged to give Z-15. MS (ESI) M/z 325[ M +1]]⁺。

Synthesis of intermediates Z-16 to Z-27

According to the synthesis method of the intermediate Z-13, the raw materials 1-c are replaced by corresponding raw material intermediate numbers, and the other methods are not changed, so that the corresponding Z-16 to Z-27 in the table are obtained.

Synthesis of intermediate Z-25

To a mixed solution of 1-c (1g,3.06mmol) in acetonitrile (66.67 mL)/water (133.33mL) was added TEMPO (191.51mg,1.23mmol) and iodobenzene diacetic acid (3.95g,12.26 mmol). The reaction mixture was stirred at room temperature overnight, after completion of the reaction, the reaction mixture was concentrated, and the crude product was isolated and purified with a normal phase silica gel column (DCM: MEOH 33:1) to give intermediate compound Z-25(248mg,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(160mg,0.47mmol) in MeOH (10mL) was added Pd/C (30 mg). The reaction solution was replaced with hydrogen, then stirred at room temperature for 1 hour under hydrogen protection, after the reaction was completed, Pd/C was filtered off, and the filtrate was spin-dried to give intermediate compound Z-26(150mg, 93% yield). MS (ESI) M/z 345[ M +1]]⁺

Synthesis of intermediate Z-27

Synthesis of step 1, 27-a

To a solution of delta-valerolactone (10g,99.88mmol) in toluene (100mL) was added BnBr (68.32g,399.54mmol) and KOH (16.81g,299.65mmol) in water (100mL) and the mixture was warmed to 115 ℃ for overnight reaction. After completion of the reaction, the reaction mixture was cooled to room temperature, 7g of KOH was added, liquid was separated, the separated aqueous phase was washed with MTBE 2 times, pH was adjusted to 2-3 with concentrated hydrochloric acid, extraction was performed with ethyl acetate, and the obtained organic phase was dried and concentrated to obtain intermediate compound 27-a (5.4g,25.93mmol, 25.96% yield), ms (esi) M/z 207[ M +1], (M + 1)]⁺

Synthesis of Steps 2, 27-b

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

Synthesis of Steps 3, 27-c

Dissolving (R) -4-benzyl-2-oxazolidinone (5.77g,32.55mmol) in THF (80mL), cooling to-78 ℃ under the protection of nitrogen, adding n-BuLi (16mL), keeping the temperature for reaction for 30 minutes, then adding a THF solution of 27-b (7.38g,32.55mmol), and keeping the temperature for reaction for 1 hour after dripping. After the reaction is finished, pouring the reaction into saturated ammonium chloride aqueous solution for quenching, separating liquid,

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

Synthesis of Steps 4, 27-d

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

Synthesis of Steps 5, 27-e

At zero degrees, the reaction was continued at a temperature of 27-d (5.4g,14.16mmol) of THF (100mL), LiAlH was added₄(15mL) and allowed to warm to 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 separation (MTBE/PE ═ 1/2) to give 27-e (2.4g,11.52mmol, 81.39% yield), ms (esi) M/z ═ 209[ M +1 ═ M +1 ═ M ═ 8 ═ M-]⁺

Synthesis of Steps 6, 27-f

27-e (2.4g,11.52mmol) was dissolved in THF (100mL) and PPh was added sequentially under nitrogen₃(6.04g,23.04mmol), DIAD (4.65g,23.04mmol) and DPPA (6.34g,23.04mmol) were stirred at room temperature overnight, after completion of the reaction, the reaction was concentrated to dryness and the residue was purified by normal phase silica gel column separation (8% MTBE in PE) to give 27-f (1.97g,8.44mmol, 73.28% yield). MS (ESI) M/z 234[ M +1]]⁺

Synthesis of Steps 7, 27-g

To 27-f (2g,8.57mmol) in MeOH (50mL) was added 2 drops of concentrated HCl followed by Pd/C (0.5 g). After hydrogen replacement, the mixture was stirred 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 (1g,8.53mmol, 99.54% yield). MS (ESI) M/z 118[ M +1]]⁺

Synthesis of Steps 8, 27-h

To a solution of 27-g (1g,8.53mmol) in DMF (20mL) was added K in sequence₂CO₃(2.36g,17.07mmol) and 2-fluoro-4-bromonitrobenzene (1.88g,8.53mmol), warmed to 60 ℃ and stirred overnight, after completion of the reaction, the reaction was concentrated and the residue was isolated and purified on normal phase silica gel column (PE/EA ═ 5/1) to give 27-h (2g,6.31mmol, 73.89% yield). MS (ESI) M/z 318[ M +1]]⁺

Synthesis of Steps 9, 27-i

To a solution of 27-h (465mg,1.47mmol) in DCM (10mL) was added TEA (222.52mg,2.20mmol,306.72 μ L) and MsCl (201.52mg,1.76mmol) in that order at zero degrees, stirred at zero degrees for 1 hour, after completion of the reaction, saturated aqueous sodium bicarbonate was added, the organic phase separated was washed with water, brine, dried over anhydrous sodium sulfate and concentrated to give 27-i (0.57g,1.44mmol, 98.36%) which was used in the next reaction without purification. MS (ESI) M/z 397[ M +1]]⁺

Synthesis of Steps 10, 27-j

27-i (579mg,1.46mmol), methyl 2- (5-hydroxy-1-methyl-1H-pyrazol-4-yl) -6-methylisonicotinate (362.18mg,1.46mmol) and K₂CO₃(505.37mg,3.66mmol) was dissolved in DMF (15mL), the reaction was allowed to warm to 60 deg.C and stirred overnight, after completion of the reaction, it was cooled to room temperature, MTBE was added, the mixture was washed with water 2 times, the separated organic phase was concentrated and the residue was purified by silica gel column separation (60% EA in PE) to give 27-j (254mg, 464.85. mu. mol, 31.73% yield). MS (ESI) M/z 548[ M +1]]⁺

Synthesis of Steps 11, 27-k

To 27-j (254mg, 464.85. mu. mol) in MeOH (10mL) at zero degrees, NH was added sequentially₄OH (3mL) and Na₂S₂O₄(404.42mg,2.32mmol) was stirred at room temperature for 1 hour, after completion of the reaction, the stone was washed with water, extracted with ethyl acetate, the combined organic phases were washed successively with water, saturated brine, dried and concentrated to give 27-k (170mg, 329.18. mu. mol, 70.81% yield) which was used in the next reaction without purification. MS (ESI) M/z 518[ M +1]]⁺

Synthesis of Steps 12, 27-l

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

Synthesis of step 13, 27-m

To 27-l (177mg, 326.91. mu. mol) of THF (5mL)/H₂To O (5mL), NaOH (52.31mg,1.31mmol) was added, the mixture was stirred at room temperature for 1 hour, after completion of the reaction, the reaction mixture was concentrated, diluted with water, adjusted to pH 4-5 with 1M HCl, and a solid precipitated, filtered, and the cake was washed with water and dried to obtain 27-M (172mg,326.12 μmol, 99.76% yield). MS (ESI) M/z 541[ M +1]]⁺

Step 14, Synthesis of Z-27

To 27-m (172mg,326.12 μmol) of DCM (5mL) was added TEA (132.00mg,1.30mmol,181.94uL) and TBTU (100.14mg,391.34 μmol) in this order, stirred at room temperature for 15 minutes, after completion of the reaction, diluted with water, separated, the organic phase concentrated, and the resulting residue was purified by silica gel column separation (DCM/MeOH ═ 10/1) to give intermediate Z-27(113mg,221.83 μmol, 68.02% yield). MS (ESI) M/z 511[ M +1]]⁺，¹H NMR(400MHz,DMSO-d₆)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

Synthesis of step 1, 28-a

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

Synthesis of step 2, 28-b

To a mixed solution of 28-a (718mg,1.12mmol) dioxane (20 mL)/water (2mL) was added potassium 4-N-Boc piperazine 1-methylboron trifluoride (515.51mg,1.68mmol), X-Phos (106.86mg, 224.50. mu. mol), Pd (OAc)₂(25.14mg, 112.25. mu. mol) and Cs₂CO₃(1.10g,3.37mmol), the mixture was replaced with nitrogen several times, then warmed to 100 ℃ under nitrogen blanket and stirred for 15 h. After completion of the reaction, the reaction solution was concentrated, and the residue was diluted with water and ethyl acetateThe combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated, and the residue was isolated and purified with normal phase silica gel column (100% EA) to give 28-b (607mg, 799.71. mu. mol, 71.25% yield). MS (ESI) M/z 759[ M +1]]⁺，¹H NMR(400MHz,DMSO-d₆)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 (671mg, 884.03. mu. mol) in DCM (5mL) was added TFA (5mL), and the reaction was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was concentrated to dryness to give Z-28(452mg, 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 steps 8 to 14 in the synthetic method of Z-27, Z-29 can be obtained by the same method except that 2-fluoro-5-bromonitrobenzene is used instead of 2-fluoro-4-bromonitrobenzene in step 8. MS (ESI) M/z 509[ M +1]]⁺。

Synthesis of intermediate Z-30

Referring to the method of step 1 to step 3 of the synthetic method of Z-28, Z-27 in step 1 is replaced with Z-29 and the remaining method steps are the same to give Z-30. MS (ESI) M/z 529[ M +1]]⁺。

Synthesis of intermediate Z-31

Step 1, Synthesis of intermediate 31-a

DIPEA (375.24mg,2.90mmol, 505.71. mu.L) and SEMCl (242.44mg,1.45mmol) were added to a solution of Z-27(493.00mg, 967.81. mu. mol) in DMF (30mL) 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 washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and after spin-drying, separated and purified (100% ethyl acetate) with a normal phase silica gel column to give 31-a (373mg, 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 (50mg, 78.17. mu. mol) in toluene (3mL) was added 1-tert-butoxycarbonylpiperazine (14.56mg, 78.17. mu. mol), BINAP (9.73mg, 15.63. mu. mol), Pd (OAc)₂(1.75mg, 7.82. mu. mol) and Cs₂CO₃(50.81mg, 156.33. mu. mol). Under nitrogen, the temperature was raised to 120 ℃ 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 separated and purified with a reverse phase silica gel column to give 31-b (30mg,40.27 μmol, 51.52% yield), ms (esi) M/z 745[ M +1]]⁺。

Step 3, synthesis of intermediate Z-31

To a solution of 31-b (30mg, 40.27. mu. mol) in DCM (3mL) was added TFA (2mL), and the mixture was stirred at room temperature for 1 h. The reaction was then concentrated and residual TFA was pumped off to give Z-31(50mg, 43.8% pure) which was used in the next reaction without purification. MS (ESI) M/z 515[ M +1]]⁺。

Synthesis of intermediate Z-32

Referring to step 1 to step 3 of the synthetic method for Z-31, Z-32 was obtained by the same method except that Z-29 was used in place of Z-27 in step 1. MS (ESI) M/z 515[ M +1]]⁺。

Synthesis of intermediate Z-33

Step 1, Synthesis of intermediate 33-a

To a solution of 4-bromo-2-methylaminoaniline (7.6g,37.8mmol) in THF (200mL) at room temperature was added CDI (7.4g,45.6 mmol). The mixture was refluxed for 16 hours, and the reaction mixture was cooled to room temperature, diluted with water (500mL) and stirred at room temperature for 30 minutes. The resulting precipitate 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 as a yellow solid, 33-a (7g, 72% yield), ms (esi) M/z 227.10,229.10[ M +1 ═ 227.10,229.10]⁺。

Step 2, Synthesis of intermediate 33-b

Under the protection of nitrogen and ice bath, 3-bromopiperidine-2, 6-dione (10g,52.08mmol), 4-methoxybenzyl alcohol (7.92g,57.29mmol) and PPh were added₃(13.66g,52.08mmol) was added dropwise to the mixture of DEAD (9.06g,52.08mmol), the reaction mixture was stirred at zero degrees for 2 hours, after completion of the reaction, water was added to quench the reaction, the mixture was extracted with ethyl acetate, the combined organic phases were concentrated to give a 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 (4.88g,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 (4.0g,17.7mmol) in THF (50mL) is cooled to 0 deg.C, t-BuOK (2.38g,21.2mmol) is added, the mixture is stirred at zero degrees for 30 minutes, then 33-b (8.26g,26.5mmol) is added, the mixture is stirred at room temperature overnight, the solvent is removed by rotation, and the crude product is purified by silica gel column separation (EtOAc: DCM-ESI 1:1) to give 33-c (2.9g, 6.33mmol, yield: 36%), MS (ESI) M/z-458 [ M +1 ESI ]]⁺。

Step 4, synthesis of intermediate Z-33

To a solution of 33-c (2.0g, 4.37mmol) in toluene (20mL) at room temperature was added MeSO₃H (10mL), the reaction mixture was warmed to 110 ℃ and stirred at this temperature for 2H. Cooling to room temperature, mixing the solutionConcentrated under reduced pressure, the crude product was diluted with ice water, and the precipitated solid was collected by filtration, washed with n-hexane and dried to give intermediate Z-33(0.8g, 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 of intermediate Z-33, step 1, 33-a, the starting material, 4-bromo-2-methylaminoaniline, was replaced with 5-bromo-2-methylaminoaniline, and the remainder was carried out unchanged to give intermediate 34-a. MS (ESI) M/z 227.10,229.10[ M +1]]⁺。

Step 2, Synthesis of intermediate 34-b

To tetrahydro-5-oxo-2-furancarboxylic acid (12g,92mmol) was slowly added SOCl at zero degrees₂(25g,207 mmol). The mixture was warmed to 85 ℃ and stirred for 3 hours, then the reaction was cooled to room temperature and stirred for 6 hours. After the raw materials completely react, the mixed solution is decompressed and concentrated. The crude product was dissolved in zero dry DCM under nitrogen and Et was added₃N (18.7g,184mmol) and 4-methoxybenzylamine (10.1g,74mmol), and the mixture was stirred at room temperature for 3 hours. After completion, the mixture was diluted with water, extracted with DCM, the combined organic phases were washed successively with 0.5M aqueous HCl (50mL) and saturated brine (50mL), dried over anhydrous sodium sulfate, filtered and spun dry, and the crude product was isolated and purified on silica gel column to give 34-b (10.5g, 45.6% yield), ms (esi) M/z 272.0[ M + Na ],]⁺。

step 3, Synthesis of intermediate 34-c

34-b (10.5g,47.9mmol) of anhydrous THF (120mL) is cooled to-78 deg.C, then t-BuOK (5.4g,48.4mmol) of anhydrous THF (100mL) is added dropwise under nitrogen protection at-78 deg.C, the reaction mixture is stirred at-40 deg.C for 1 hour, after the reaction is completed, the mixture is saturated NH₄Quenching with Cl solution, extracting with ethyl acetate, washing the combined organic phase with saturated sodium chloride water, drying with anhydrous sodium sulfate, filtering, concentrating, and separating with silica gel columnThe reaction yielded intermediate 34-c (10.9g,43.73mmol, 77.7% yield). MS (ESI) M/z 250.0[ M +1]]⁺。

Step 4, Synthesis of intermediate 34-d

To a solution of 34-c (10.9g,43.73mmol) and pyridine (6.92g,87.46mmol) in DCM (100mL) was added dropwise trifluoromethanesulfonic anhydride (18.51g,65.59mmol) at zero degrees, and the mixture was stirred at 0-10 deg.C for 1.5 h under nitrogen. After completion of the reaction, the mixture was concentrated and the crude product was purified by silica gel column separation to give 34-d (8.17g,21.43mmol, 49% yield), ms (esi) M/z ═ 382[ M +1 ═ 382]⁺。

Step 5, Synthesis of intermediate 34-e

t-BuOK (2.89g,25.7mmol) was added to a solution of 34-a (4.86g,21.43mmol) in THF (100mL) at zero degrees, the mixture was stirred at 0 ℃ for 0.5h, then a solution of 34-d (8.17g,21.43mmol) in THF (50mL) was added dropwise, and the reaction was stirred at room temperature under nitrogen for 0.5 h. After the reaction is finished, saturated NH is used for reaction liquid₄After quenching with Cl (40mL), extraction with ethyl acetate and washing of the combined organic phases with saturated brine, drying over anhydrous sodium sulfate, filtration and concentration, the crude product was isolated and purified by mHPLC to give 34-e (4.91g,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, the same procedure was followed except that the substrate 33-c in this step was changed to 34-e, to give intermediate Z-34, MS (ESI) M/Z338,340 [ M +1] M]⁺。

Synthesis of intermediate Z-35

Referring to the synthetic route for intermediate Z-33, the procedure of steps 1 to 4, starting from 3-bromo-2-methylaminoaniline instead of 4-bromo-2-methylaminoaniline in step 1, was followed in the same manner as the rest of the procedure, to give intermediate Z-35, ms (esi) M/Z338,340 [ M +1] M]⁺。

Synthesis of intermediate Z-36

Step 1, Synthesis of intermediate 36-a

4-Pyrazoleboronic acid pinacol ester (5g,25.77mmol) and 3-benzyloxybromopropane (8.86g,38.65mmol) were mixed in DMF (20mL) solution, and K was added₂CO₃(8.90g,64.42mmol), KI (427.76mg,2.58 mmol). The reaction mixture was warmed to 80 ℃ and stirred for 17 hours. After completion of the reaction, the crude product obtained from the reaction mixture was concentrated and purified by silica gel column separation (PE: EA ═ 8:1, v/v) to obtain intermediate 36-a (6.4g,18.70mmol, 72.57% yield), ms (esi) M/z ═ 343[ M + H ═ M + H]⁺。

Step 2, Synthesis of intermediate 36-b

36-a (2g,5.84mmol) and 1-b (1.89g,5.84mmol) were mixed in DMF (20mL) and Na was added₂CO₃(1.86g,17.53mmol) and Pd (dppf) Cl₂(213.80mg,292.19umol), the mixture was purged with nitrogen several times, then warmed to 100 ℃ under nitrogen, and stirred overnight. After completion of the reaction, it was cooled to room temperature, diluted with water, extracted with ethyl acetate, the combined organic phases were washed with saturated brine, concentrated and the crude product was purified by silica gel column separation (100% EA) to give intermediate 36-b (1.6g,3.49mmol, 59.71% yield), ms (esi) M/z ═ 459[ M + H ], (M + H)]⁺。

Step 3, Synthesis of intermediate 36-c

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

Step 4, synthesis of intermediate Z-36

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

Synthesis of intermediate Z-37

Step 1, Synthesis of intermediate 37-a

To 4-pyrazole boronic acid pinacol ester (6.0g,30.92mmol) in acetonitrile (60mL) were added CsF (470mg,3.09mmol) and tert-butyl acrylate (5.82mL,40.2 mmol). The reaction mixture was warmed to 80 ℃ and stirred at this temperature overnight. After completion of the reaction, it was cooled to room temperature, concentrated under reduced pressure, and the crude product was extracted with water and ethyl acetate, and the separated organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was purified by silica gel column separation (n-hexane/ethyl acetate ═ 1/1, v/v) to obtain 37-a (9.37g,29.07mmol, yield 94%). MS (ESI) M/z 323[ M + H ]]⁺。

Step 2, Synthesis of intermediate 37-b

To 37-a (500mg,1.55mmol) and 1-b (455.86mg,1.41mmol) in DMF (12mL)/H₂Adding K into O (3mL) solution₃PO₄(416.73mg,1.98mmol), S-phos (69.41mg, 169.29. mu. mol) and Pd (OAc)₂(22.61mg, 98.75. mu. mol), the mixture was purged with nitrogen, heated to 100 ℃ under nitrogen protection, and stirred at this temperature for 15 hours to complete the reaction. Cooling to room temperature, filtering, concentrating the filtrate, diluting the crude product with water, extracting with ethyl acetate, washing the combined organic phases with water and saturated brine, drying over anhydrous sodium sulfate, concentrating, and isolating and purifying the crude product with silica gel column (PE/EA ═ 0/1, v/v) to give 37-b (100mg,228.06 μmol, 16.17% yield) as a pale yellow solid. MS (ESI) 439[ M + H ] M/z]⁺。

Step 3, synthesis of intermediate Z-37

To a solution of 37-b (100mg, 228.06. mu. mol) in DCM (2mL) was added TFA (1.54g,13.51mmol,1mL) at room temperature, and the mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction mixture was concentrated and dried to obtain Z-37(92mg, 185.34. mu. mol, 81.27% yield), a crude productPurified and directly used for the next reaction. MS (ESI) M/z 383[ M + H ]]⁺。

Synthesis of intermediate Z-38

Step 1, Synthesis of intermediate 38-a

To a solution of methyl 2-methyl 5-iodobenzoate (10g,36.22mmol) in CCl₄NBS (6.77g,38.03mmol) and benzoyl peroxide (4.38g,18.11mmol) were added to the solution (100mL), and the mixture was warmed to 80 ℃ and stirred for 4 hours. After completion of the reaction, it was cooled to room temperature, filtered, and the filtrate was concentrated to give 38-a (12g,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 (12g,33.81mmol) in DMF (150mL) was added 3-amino-2, 6-piperidinedione hydrochloride (6.12g,37.19mmol) and K₂CO₃(14.02g,101.42 mmol). The reaction mixture was warmed to 70 ℃ and stirred for 18 h. After the reaction was completed, the reaction solution was cooled to room temperature, diluted with water, stirred for 10min, filtered, the filter cake was washed with petroleum ether and ethyl acetate, the filter cake was collected and dried to obtain 38-b (6.5g,17.56mmol, 51.95% yield)]⁺。

Step 3, Synthesis of intermediate 38-c

To a solution of 38-b (400mg,1.08mmol) and tert-butyl 2- (4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazol-1-yl) acetate (366.34mg,1.19mmol) in dioxane (5mL)/H2O (0.5mL) was added K₃PO₄(320.74mg,1.51mmol), S-phos (53.17mg, 129.68. mu. mol) and Pd (OAc)₂(17.32mg, 75.65. mu. mol), the mixture was purged with nitrogen several times, then warmed to 100 ℃ under nitrogen atmosphere, and stirred overnight. After the reaction is finished, cooling to room temperature, adding water for dilution, adding ethyl acetate for extraction, washing combined organic phases with water and saturated saline solution, drying with anhydrous sodium sulfate, concentrating a crude product, and performing DMF/H (dimethyl formamide/hydrogen) treatment on the crude product₂Recrystallization of the mixture of O gave 38-c (200mg, 471.20. mumol, 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 (200mg, 471.20. mu. mol) in DCM (3mL) was added TFA (53.73mg, 471.20. mu. mol, 35.12. mu.L) at room temperature, and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction, the reaction was concentrated and the crude product was dried to give Z-38(200mg, 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 (1g,2.70mmol) and 36-a (1.02g,2.97mmol) in dioxane (20mL)/H₂Na was added to the O (1mL) mixture₂CO₃(400.93mg,3.78mmol), S-phos (132.92mg, 324.20. mu. mol) and Pd (OAc)₂(43.31mg, 189.12. mu. mol), the mixture was purged with nitrogen several times, and the reaction mixture was heated to 60 ℃ under nitrogen protection, and stirred at 60 ℃ for overnight reaction. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, the obtained crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and after spin-drying, the crude product was separated and purified by pre-HPLC to give intermediate 39-a (130mg, 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 (1.09g, 894.20. mu. mol, 10% purity) was added to a mixture of 39-a (410mg, 894.20. mu. mol) in MeOH (150mL) and THF (30mL), the reaction solution was replaced several times with hydrogen, and then stirred at room temperature for 15h under protection of hydrogen, after completion of the reaction, filtration was carried out, and the filtrate was concentrated and dried to give the crude intermediate 39-b (310mg) which was used in the next reaction without purification.

Step 3, Synthesis of intermediate Z-39

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

Synthesis of intermediate Z-40

Step 1, Synthesis of intermediate 40-a

To a solution of 2-b (10.0g,30.9mmol) and DBU (10.4mL,69mmol) in DMF (95mL) under ice-bath was added SEMCl (9.9mL,56mmol), the mixture was slowly warmed to room temperature and the reaction was stirred for 6 h. After the reaction is finished, saturated NH is used for reaction liquid₄Quenching with Cl (250mL), extracting with ethyl acetate, drying the combined organic phases with anhydrous sodium sulfate, filtering, concentrating the filtrate to obtain a crude product, and recrystallizing the crude product with a mixture of ethyl acetate/diethyl ether/n-hexane to obtain intermediate 40-a (9.5g,30.95mmol, 67.7% yield) as a white solid.

Step 2, Synthesis of intermediate 40-b

To 40-a (1g,2.21mmol) and 36-a (830.31mg,2.43mmol) in dioxane (5.00mL)/H₂Na was added to the O (5.00mL) mixture₂CO₃(327.30mg,3.09mmol), S-phos (108.51mg, 264.67. mu. mol) and Pd (OAc)₂(35.35mg,154.39 μmol), replacing the mixture with nitrogen several times, heating to 60 ℃ under the protection of nitrogen, stirring at the temperature for 15 hours, filtering the reaction solution after the reaction is finished, concentrating the filtrate, adding water to dilute the obtained crude product, extracting with ethyl acetate, washing the combined organic phases with water and saturated saline, drying with anhydrous sodium sulfate, and separating and purifying the crude product by a silica gel column (PE/EA 1/1) to obtain an intermediate 40-b (200mg,339.69 μmol, 15.40% yield) as a white solid. MS (ESI) M/z 589[ M + H ]]⁺。

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

According to the method of step 2 and step 3 of the intermediate Z-39 synthesis method, 40-b is taken as raw material and is Pd/based onC reduction and DMP oxidation to obtain intermediate Z-40, MS (ESI) M/Z497 [ M + H ]]⁺。

Synthesis of intermediate Z-41

Step 1, Synthesis of intermediate 41-a

To intermediate 40-a (469.02mg,1.46mmol) and 37-a (600mg,1.32mmol) in DMF (8mL)/H₂Adding K into O (2mL) mixed solution₃PO₄(393.32mg,1.85mmol), S-phos (65.19mg, 159.01. mu. mol) and Pd (OAc)₂(21.21mg, 92.63. mu. mol), the mixture was purged with nitrogen several times, heated to 100 ℃ under nitrogen, and the reaction was stirred at this temperature for 15 hours. After 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, and 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 silica gel column (PE/EA ═ 2/1) to obtain intermediate 41-a (130mg,228.58 μmol, 17.27% yield) as a white solid. MS (ESI) M/z 383[ M + H ]]⁺。

Step 2, synthesis of intermediate Z-41

TFA (18.04mg, 158.25. mu. mol) was added to a solution of 41-a (90mg, 158.25. mu. mol) in DCM (2mL) at room temperature, the mixture was stirred at room temperature for 2 hours, after completion of the reaction, the reaction was concentrated, and the crude product was redissolved in DCM (5mL), followed by TEA (16.01mg, 158.25. mu. mol, 22.07. mu.L) at room temperature and stirred at room temperature for 30 minutes. The reaction was again concentrated and purified by pre-HPLC to give intermediate Z-41(50mg, 130.76. mu. mol, 82.63% yield) as a white solid.

Synthesis of intermediate Z-42

Step 1, Synthesis of intermediate 42-a

To 3- (4-hydroxy-1-oxo-1, 3-dihydroisoindol-2-yl) piperidine-2, 6-dione (500mg,1.92mmol) in DMF (8mL)) To the solution was added 4-bromobutyloxy-tert-butyldimethylsilane (514mg,1.92mmol), KHCO₃(570mg,5.76mmol) and catalytic amount of KI, the mixture was warmed to 70 ℃ and the reaction stirred at this temperature for 15 h. After the reaction, the reaction solution was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered and dried to give a crude product, which was separated and purified by means of hplc to give intermediate 42-a (160mg,0.48mmol, 25% yield). MS (ESI) M/z 333[ M + H ]]⁺。

Step 2, synthesis of intermediate Z-42

DMP (204.12mg, 481.42. mu. mol) was added to a solution of 42-a (80mg, 240.71. mu. mol) in DCM (10mL), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was filtered, the filtrate was concentrated, and the crude product was isolated and purified by silica gel column (DCM/MeOH ═ 10/1) to give intermediate Z-42(75mg,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 (5.0g,50.94mmol) is dissolved in DMF (100mL), the temperature is reduced to 0 ℃ under the protection of nitrogen, NaH (2.04g,50.94mmol) is added, the reaction is kept for 30 minutes, 3-benzyloxy bromopropane (9.34g,40.76mmol) is added, the reaction mixture is heated to 25 ℃ and stirred for 2 hours, after the reaction is finished, saturated ammonium chloride aqueous solution is added for quenching, water is added for dilution, EA extraction is carried out, the combined organic phase is washed by water and saturated common salt water, anhydrous sodium sulfate is dried, and separation and purification are carried out by silica gel column (PE/EA is 20/1) after spin drying, so that 43-b (2.5g,10.15mmol, 19.92% yield) is obtained. MS (ESI) M/z 247[ M +1]]⁺。

Step 2, Synthesis of intermediate 43-c

Dissolving 1-b (20.0g,61.89mmol) in DMF (500mL), cooling to 0 deg.C under nitrogen protection, adding NaH (2.72g,68.08mmol), reacting for 30min under heat preservation, adding SEMCl (12.66g,75.83mmol), heating the reaction mixture to 25 deg.CStirring for 12h, after the reaction is finished, adding saturated ammonium chloride aqueous solution for quenching, adding water for diluting, extracting with EA, washing the combined organic phase with water and saturated saline solution successively, drying with anhydrous sodium sulfate, and spin-drying to obtain crude product 43-c (28g,61.76mmol, 99.78% yield). MS (ESI) M/z 453[ M +1]]⁺And directly used in the next step.

Step 3, Synthesis of intermediate 43-d

To a solution of 43-c (6.91g,15.24mmol) in DMF (100mL) was added 43-b (4.13g,16.76mmol), CuI (435.38mg,2.29mmol), Pd (Ph) in that order₃P)₂Cl₂(801.24mg,1.14mmol) and TEA (4.63g,45.72mmol,6.37 mL). Reaction solution in N₂Stirring at 85 deg.C for 12 hr under protection, diluting with water, extracting with EA, washing the separated organic phase with water and brine, drying, and spin-drying. The crude product was purified by silica gel column separation (PE/EA ═ 1/1) to give intermediate 43-d (6.66g,10.76mmol, 70.62% yield). MS (ESI) M/z 619[ M + H ]]⁺。

Step 4, synthesis of intermediate 43-e

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

Step 5, Synthesis of intermediate Z-43

DMP (238.76mg, 563.12. mu. mol) was added to a solution of 43-e (200mg, 375.42. mu. mol) in DCM (10mL), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was filtered, the filtrate was concentrated, and the crude product was isolated and 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

In the synthetic route referring to intermediate Z-43,the process of step 1 to step 5, wherein 4-pentynyl-1-ol is used as the starting material in step 1 instead of 5-hexynyl-1-ol aniline, and the same procedure is followed to give intermediate Z-44, ms (esi) M/Z517 [ M +1 ═ M]⁺。

Synthesis of intermediate Z-45

Step 1, Synthesis of intermediate 45-b

To 28-a (50mg, 78.17. mu. mol) in DMF (5mL)/H₂To a solution of O (0.5mL) were added 45-a (24.17mg,78.17mmol), K in that order₂CO₃(32.36mg,234.5μmol)，Pd(dppf)Cl₂(5.71mg, 7.82. mu. mol). Reaction solution in N₂Stirring at 90 deg.C for 12 hr under protection, diluting with water, extracting with ethyl acetate, washing the separated organic phase with water and brine, drying, and spin-drying. The crude product was purified by pre-TLC (100% EA) to give intermediate 45-b (57mg,76.82mmol, 98.28% yield). MS (ESI) M/z 742[ M + H ]]⁺。

Step 2, synthesis of intermediate Z-45

TFA (18.04mg, 158.25. mu. mol) was added to a solution of 45-b (32mg, 43.13. mu. mol) in DCM (2mL) at room temperature, the mixture was stirred at room temperature for 2 hours, and after completion of the reaction, the reaction was concentrated to give crude intermediate Z-45(22mg, 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

Adding Pd/C (3mg, 10% purity) into 45-b (32mg, 43.13. mu. mol) MeOH (5mL), replacing the reaction solution with hydrogen for several times, stirring at room temperature for 12h under the protection of hydrogen, filtering after the reaction is finished, concentrating and drying the filtrate to obtain crude intermediate 46-a (32mg, 43.01. mu. mol, 99.73% yield), and directly using the crude intermediate in the next step without purificationAnd (5) carrying out reaction. MS (ESI) M/z 744[ M + H ]]⁺。

Step 2, synthesis of intermediate Z-46

TFA (18.04mg, 158.25. mu. mol) was added to a solution of 46-a (33mg, 44.35. mu. mol) in DCM (2mL) at room temperature, the mixture was stirred at room temperature for 2 hours, and after completion of the reaction, the reaction was concentrated to give crude intermediate Z-46(22mg, 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 (200mg, 431.35. mu. mol) in DCM (10mL) was added DMP (267.98mg, 632.03. mu. mol), and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was filtered, the filtrate was concentrated, and the crude product was isolated and purified by silica gel column (PE/EA ═ 1/2), yielding 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

To a solution of 48-a (200mg, 724.06. mu. mol) in NMP (10mL) was added 48-b (100.07mg, 838.87. mu. mol), DIPEA (280.73mg,2.17mmol, 378.35. mu.L), and the mixture was heated to 90 ℃ under nitrogen for 12 hours. After completion of the reaction, water was added, EA extracted, the organic phase was concentrated and the crude product was purified by silica gel column separation (PE/EA ═ 1/4) to give intermediate 48-c (200mg,538.52 μmol, 74.38% yield). MS (ESI) M/z 372[ M + H ]]⁺。

Step 2, synthesis of intermediate Z-48

DMP (342.50mg, 807.78. mu. mol) was added to a solution of 48-c (200mg, 538.52. mu. mol) in DCM (10mL), and the mixture was stirred at room temperature for 1 hour. After the reaction is completed, the reaction solution is filtered, and the filtrate is concentratedThe crude product was purified by silica gel column separation (PE/EA ═ 1/4) to give intermediate Z-48(134mg,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

Dissolving 2-b (3.0g,9.28mmol) in DMF (50mL), cooling to 0 ℃ under nitrogen, adding NaH (445.63mg,11.14mmol), keeping the temperature for reaction for 30 minutes, adding SEMCl (1.86g,11.14mmol), heating the reaction mixture to 25 ℃ and stirring for 12 hours, after the reaction is completed, adding saturated aqueous ammonium chloride solution for quenching, adding water for dilution, extracting with ethyl acetate, washing the combined organic phase with water and saturated brine successively, drying over anhydrous sodium sulfate, spin-drying, separating and purifying the crude product with silica gel column (PE/EA ═ 1/1), to obtain intermediate 49-a (2.97g,6.55mmol, 70.56% yield). MS (ESI) M/z 453[ M +1]]⁺。

Step 2, Synthesis of intermediate 49-c

49-a (0.2g, 441.11. mu. mol), 49-b (127.97mg, 529.33. mu. mol), Pd (OAc)₂(3.96mg,17.64μmol)，BINAP(21.97mg,35.29μmol)，Cs₂CO₃(286.72mg, 882.22. mu. mol) was dissolved in a mixed solvent of toluene (10mL) and t-BuOH (1mL), and the mixture was heated to 100 ℃ for 12 hours after nitrogen substitution. After completion of the reaction, water was added, EA was extracted, the organic phase was washed successively with water, saturated brine, dried over anhydrous sodium sulfate, spun-dried, and the crude product was isolated and purified by silica gel column (100% EA) to obtain intermediate 49-c (40mg, 69.23. mu. mol, 15.69% yield). MS (ESI) M/z 578[ M +1]]⁺。

Step 3, Synthesis of intermediate 49-d

Pd/C (5mg, 10% purity) was added to a solution of 49-C (45mg, 77.88. mu. mol) in MeOH (5mL), the reaction was replaced several times with hydrogen, then stirred at room temperature for 4h under protection of hydrogen, after completion of the reaction, filtered, and the filtrate was concentrated and dried to give a crude intermediate 49-d (36mg, 73.82. mu. mol, 94.87% yield), which was used directly in the next step without purificationAnd (4) carrying out one-step reaction. MS (ESI) M/z 488[ M + H ]]⁺。

Step 4, synthesis of intermediate Z-49

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

Synthesis of intermediate Z-50

Synthesis of step 1, 50-a

To 27-g (3.0g,25.64mmol) of THF (100mL)/H at zero degrees₂To a solution of O (50mL), potassium carbonate (10.62g,76.92mmol) and Cbz-OSu (12.82g,51.28mmol) were added in this order, and the mixture was stirred at room temperature for 1 hour, after completion of the reaction, a saturated aqueous solution of sodium hydrogencarbonate was added, EA was added for extraction and liquid separation, the separated organic phase was washed with water, saturated brine, dried over anhydrous sodium sulfate and concentrated, and the residue was purified by silica gel column separation (50% EA in PE) to obtain 50-a (5.1g,20.29mmol, 79.13% yield). MS (ESI) M/z 252[ M +1]]⁺

Synthesis of Steps 2, 50-b

To a solution of 50-a (5.1g,20.29mmol) in DCM (100mL) was added TEA (6.16g,60.88mmol,8.49mL) and MsCl (3.49g,30.44mmol) sequentially at zero degrees, stirred at zero degrees for 1 hour, after completion of the reaction, a saturated aqueous sodium bicarbonate solution was added, separated, the separated organic phase was washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give 50-b (6.66g,20.22mmol, 99.63% yield), which was used in the next reaction without purification. MS (ESI) M/z 330[ M +1]]⁺

Synthesis of Steps 3, 50-d

50-b (6.68g,20.28mmol), 50-c (5.52g,22.31mmol), NaI (303.96mg, 2.03mmol) and K₂CO₃(8.40g,60.84mmol) was dissolved in DMF (100mL) and the reaction was raisedAfter completion of the reaction, the reaction was cooled to room temperature, EA was added, the mixture was washed with water 2 times, the separated organic phase was concentrated, and the residue was purified by silica gel column separation (50% EA in PE) to give 50-d (3.3g,6.87mmol, 33.86% yield). MS (ESI) M/z 481[ M +1]]⁺

Step 4, synthesis of intermediate 50-e

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

Step 5, Synthesis of 50-g

After the completion of the reaction by adding DIPEA (2.42g,18.71mmol) and 50-f (1.20g,6.24mmol) in this order to a solution of 50-e (2.16g,6.24mmol) in DCM (50mL) under ice-cooling and stirring overnight at room temperature, the reaction mixture was concentrated, and the residue was purified by normal phase silica gel column separation (PE/EA ═ 1/1) to obtain 50-g (3.0g,5.96mmol, 95.66% yield). MS (ESI) M/z 503[ M +1]]⁺

Step 6, Synthesis of 50-h

To a solution of 50-g (3.82g,7.60mmol) of EtOH (60mL)/H₂To O (10mL), NH was added in order₄Cl (4.03g, 75.95mmol) and Fe (2.55g,45.57mmol) were stirred at 80 ℃ for 12 hours, after the reaction was complete, filtration was carried out, the filtrate was rinsed with EA and concentrated to give 50-h (3.15g,6.66mmol, 87.69% yield), the crude product was used in the next reaction without purification. MS (ESI) M/z 473[ M +1]]⁺

Synthesis of Steps 7, 50-i

To a 50-h (100mg, 211.43. mu. mol) mixed solution in EtOH (3mL) was added CNBr (111.06mg,1.06mmol) and the mixture was stirred at 60 ℃ overnight. After completion of the reaction, a saturated aqueous sodium bicarbonate solution was added, and the mixture was stirred for 10 minutes, separated, and the organic phase was washed with a saturated aqueous sodium bicarbonate solution, water, and brine, in that order, dried over anhydrous sodium sulfate, concentrated, and the resulting residue was purified by silica gel column separation (DCM/MeOH 10/1) to obtain 50-i (90mg,180.73 μmol, 85.48% yield). MS (ESI) M/z 498[ M +1]⁺

Synthesis of step 8, 50-j

To 50-i (2.0g,4.02mmol) in THF (50mL)/H₂To O (50mL), NaOH (642.60mg,16.07mmol) was added, and the mixture was stirred at room temperature for 1 hour, after completion of the reaction, the reaction mixture was concentrated, diluted with water, adjusted to pH 4-5 with 1M HCl, and a solid was precipitated, filtered, and the cake was washed with water and dried to obtain 50-j (1.2g,2.48mmol, 61.74% yield). MS (ESI) M/z 484[ M +1]]⁺。

Synthesis of Steps 9, 50-k

To 50-j (1.23g,2.54mmol) of DCM (5mL) was added TEA (1.03g,10.17mmol,1.42mL) and TBTU (979.02mg,3.05mmol) in that order, stirred at room temperature for 15 minutes, 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 (1.0g,2.15mmol, 84.44% yield). MS (ESI) M/z 466[ M +1]]⁺。

Step 10, Synthesis of intermediate 50-l

DIPEA (832.12mg,6.44mmol,1.12mL) and SEMCl (358.42mg,2.15mmol) were added sequentially to a 50-k (1.0g,2.15mmol) solution in DMF (30mL), 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, spin-dried and purified by normal phase silica gel column separation (100% ethyl acetate) to obtain 50-l (630mg,1.06mmol, 49.24% yield). MS (ESI) M/z 596[ M +1]]⁺。

Step 11, Synthesis of intermediate 50-m

To a 50-l (300mg, 503.19. mu. mol) solution in toluene (4 mL)/tert-butanol (1mL) was added 1-tert-butoxycarbonylpiperazine (140.58mg, 754.79. mu. mol), BINAP (25.07mg, 40.26. mu. mol), Pd (OAc)₂(4.51mg, 20.13. mu. mol) and Cs₂CO₃(327.07mg,1.01 mmol). Under nitrogen, the temperature was raised to 120 ℃ 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 separated and purified with reverse phase silica gel column to give 50-M (370mg,495.99 μmol, 98.57% yield), ms (esi) M/z 746[ M +1]]⁺。

Step 12, Synthesis of intermediate Z-50

To a solution of 50-m (390mg, 522.80. mu. mol) in DCM (10mL) was added TFA (10mL), and the mixture was stirred at room temperature for 1 h. The reaction was then concentrated and residual TFA was pumped off to give Z-50(269mg, 521.71. mu. mol, 99.79% yield) which 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

To a solution of 1-b (16g,49.51mmol) in DMF (470mL) at zero degrees was added NaH (1.43g,59.42mmol), the mixture was stirred at zero degrees for 1 hour, then SEMCl (9.08g,54.47mmol) was added. After the dripping is finished, the mixed solution reacts for 12 hours at room temperature. After completion of the reaction, water was added for dilution, EtOAc was added for extraction, and the combined organic phases were washed with water, brine, dried over anhydrous sodium sulfate and concentrated to give crude 51-a (22g,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 synthetic route of intermediate Z-1, the procedures of steps 3, 4 and 5, intermediate 51-a was used instead of intermediate 1-b, and the remaining procedures were the same, giving intermediate Z-51. MS (ESI) M/z 445[ M +1]]⁺。

Synthesis of intermediate Z-52

Referring to the synthesis of intermediate Z-42, intermediate Z-52 was obtained by the same procedure as described above except that 4-bromopentyloxy-tert-butyldimethylsilane was replaced with 4-bromopentyloxy-tert-butyldimethylsilane in step 1. MS (ESI) M/z 345[ M +1]]⁺。

Synthesis of intermediate Z-53

Referring to the procedure for steps 11 and 12 of the intermediate Z-50 synthetic route, the same procedure was followed using 1-Boc-4- (piperidin-4-yl) -piperazine instead of 1-tert-butoxycarbonylpiperazine in step 11 to give intermediate Z-53, ms (esi) M/Z599 [ M +1] M]⁺。

Synthesis of intermediate Z-54

Referring to the procedure for steps 11 and 12 of the intermediate Z-50 synthetic route, substituting intermediate 28-a for intermediate 50-l in step 11 and 4-piperazin-1-ylpiperidine-1-carboxylic acid tert-butyl ester for 1-tert-butoxycarbonylpiperazine with the same procedure as the rest, intermediate Z-54 is obtained, ms (esi) M/Z598 [ M +1 ═ M]⁺。

Synthesis of intermediate Z-55

Referring to the procedures from step 5 to step 12 in the route of intermediate Z-50, 2, 6-dichloro-3-nitropyridine was replaced with 2, 4-dichloro-5-nitropyridine in step 5, and 1-tert-butoxycarbonylpiperazine was replaced with tert-butyl 4-piperazin-1-ylpiperidine-1-carboxylate in step 11, and the rest of the procedures were the same, giving intermediate Z-55. MS (ESI) M/z 598[ M +1]]⁺。

Synthesis of intermediate Z-56

Referring to the procedures of step 11 and step 12 in the synthetic route for intermediate Z-50, intermediate Z-56 can be obtained by replacing 1-tert-butoxycarbonylpiperazine with tert-butyl 4-piperazin-1-ylpiperidine-1-carboxylate in step 11 and the same procedure. MS (ESI) M/z 599[ M +1]]⁺。

Synthesis of intermediate Z-57

Referring to the procedures of step 11 and step 12 in the synthetic route for intermediate Z-50, intermediate Z-57 can be obtained by replacing intermediate 50-l with intermediate 28-a and 1-t-butoxycarbonylpiperazine with t-butyl 4- [ (piperazin-1-yl) methyl ] piperidine-1-carboxylate, and the remaining procedures were the same. Ms (esi) M/z 612[ M +1] +.

Synthesis of intermediate Z-58

Referring to the procedures of step 7 and step 8 in the synthetic route of intermediate Z-55, the same procedure was followed except that 1-tert-butoxycarbonylpiperazine was used instead of tert-butyl 4-piperazin-1-ylpiperidine-1-carboxylate, to give 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 59-a was obtained by substituting 4-c for intermediate 1-c, and the same operating conditions were used.

Step 2, synthesis of intermediate Z-59

By referring to the synthesis method of intermediate Z-26, starting from 59-a instead of Z-25, the same operations and conditions can give intermediate Z-59.

Synthesis of intermediate Z-60

Reference intermediate Z-43The procedure outlined for steps 1 through 4, substituting 43-a with 7 octynol in step 1 as the starting material, and the remaining procedures were the same, to give intermediate Z-60, MS (ESI) M/Z ═ 559[ M +1] 559]⁺。

Synthesis of intermediate Z-61

Referring to the procedure of step 2 to step 4 of the synthetic scheme of example Z-1, the 4-pentyn-1-ol was replaced with 10-undecenol in the initial step and the remaining procedures were 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 synthetic route for intermediate Z-50, the 1-tert-butoxycarbonylpiperazine was replaced with tert-butyl 3- (piperazin-1-ylmethyl) pyrrolidine-1-carboxylate in the initial step, and the remaining procedures were the same, giving intermediate Z-62. MS (ESI) M/z 599[ M +1]]⁺。

Synthesis of intermediate Z-63

Step 1

Z-63-1a (2g, 9.2mmol) and pyrazole (755mg, 11mmol) are dissolved in acetonitrile (20ml), acetyl chloride (872mg, 11mmol) is added dropwise after cooling in an ice water bath (0 ℃ -5 ℃), then the mixture reacts at room temperature for 17h, Z-63-1b (1.5g, 9.2mmol), pyrazole (1.4g, 19.3mmol) and triphenyl phosphite (3.4g, 11mmol) are added directly into the reaction solution the next day, then the reaction solution is heated to 85 ℃ for 17h, after the reaction is completed, the reaction solution is cooled to room temperature, water is added into the reaction solution, solid is separated out, the solid is filtered and washed with water for 1 time, ethyl acetate is washed for 1 time, then the solid is filtered and dried in a rotary mode, and is directly used for the next step.

Step 2

Z-63-1c (1.58g, 4.5mmol) is dissolved in DMF, cooled in an ice-water bath (0 ℃ to 5 ℃), added with sodium hydride (130mg, 5.4mmol), kept at the temperature for reaction for 1 hour, then added with SEMCl (906mg, 5.4mmol) slowly in drops, warmed to room temperature for reaction for 3 hours, added with water after the reaction is finished, extracted with ethyl acetate for 3 times, washed once with saturated common salt water of the organic layer, dried over anhydrous sodium sulfate, concentrated and used directly in the next step.

Step 3

Z-63-1d (500mg, 1.04mmol) and 4-pentyn-1-ol (105mg, 1.25mmol) were dissolved in DMF and Pd (pph) was added₃)₂Cl₂(55mg, 0.078mmol), CuI (30mg, 0.156mmol) and TEA (315mg, 3.12mmol) were heated to 80 ℃ to react for 17 hours, after completion of the reaction, water was added, EA was added for extraction and separation, the separated organic phase was washed successively with water, saturated brine, dried over anhydrous sodium sulfate and concentrated, and the crude product was isolated and purified by silica gel column separation (50% EA in PE) to give Z-63-1e (240mg, 0.496mmol, 47.8% yield). MS (ESI) M/z 484[ M +1]]⁺

Step 4

Z-63-1e (240mg, 0.496mmol) was dissolved in methanol, 10% palladium on carbon (35mg) was added and the reaction was carried out at room temperature for 17h under hydrogen protection, after completion of the reaction, filtration was carried out and the filtrate was dried by spinning and used in the next step as it was to give Z-63-1f (220mg, 0.451mmol, 90.1% yield) MS (ESI) M/Z ═ 488[ M +1 ═ M]⁺

Step 5

Z-63-1f (220mg, 0.451mmol) was dissolved in dichloromethane, cooled in an ice bath, DMP (229mg, 0.54mmol) was added and then allowed to warm to room temperature for 4h, after completion of the reaction the solid was filtered off, the filtrate was dried and the crude product was purified on column (PE: EA ═ 2:1) to give compound Z-63(120mg, 0.246mmol, 52% yield), ms (esi) M/Z ═ 486[ M +1 ═ 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 (1.57g, 5.7mmol) in DMF (50mL) was added potassium carbonate (1.19g, 8.58mmol) and tert-butyl bromoacetate (1.1g, 5.71 mmol). After reacting for 2h at room temperature, the DMF is removed by rotary evaporation, the ethyl acetate is dissolved and then washed by water, and the organic phase is dried by rotary drying. The crude product was purified by silica gel column (petroleum ether/ethyl acetate. cndot. ═ 2/1) to give intermediate 2- (2, 6-dioxo-piperidin-3-yl) -4-hydroxyacetic acid tert-butyl ester-isoindole-1, 3-dione (2g, 5.1 mmol). This was dissolved in methylene chloride (15mL), and trifluoroacetic acid (10mL) was added to react at room temperature for 1h, followed by spin-drying of 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 initial raw material 2-amino-6-bromobenzoic acid is replaced by 2-amino-5-bromobenzoic acid, and other reagents and operation methods are the same, so that an intermediate Z-65 is obtained.

Synthesis of intermediate Z-66

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

Synthesis of intermediate Z-67

Referring to the procedure of step 1 to step 4 of the synthetic route for intermediate Z-36, 3-benzyloxybromopropane was replaced with 2-benzyloxybromoethane in the initial step and the same procedure was followed to give intermediate Z-67. MS (ESI) M/z 353M +1]⁺。

Synthesis of intermediate Z-68

Referring to the synthesis method of intermediate Z-25, intermediate Z-68 can be obtained by replacing 1-c with 3-c and performing the same procedures with the other reagents. MS (ESI) M/z 341[ M +1]]⁺。

Example 1 Synthesis of Compound 1

To Z-28(30mg,56.75 μmol) in MeOH (2mL) was added TEA to pH 9-10, followed by Z-2(18.63mg,56.75 μmol), stirred at rt for 30min, and then adjusted to pH 4-5 with acetic acid. After stirring for 5 minutes, NaBH was added in one portion₃CN (10.73mg, 170.25. mu. mol), the reaction mixture was left 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(1.42mg,1.59 μmol, 2.80% yield, 94% purity). MS (ESI) M/z 837[ M +1]]⁺，¹H NMR(400MHz,DMSO-d₆)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

Z-28(46.60mg, 88.15. mu. mol) and Z-25(30mg, 88.15. mu. mol) were mixed in DMF (5mL) and DIPEA (56.96mg, 440.75. mu. mol, 76.77. mu.L) and HATU (40.22mg, 105.78. mu. mol) were 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 subjected to HPLC separation and purification to obtain the productExample 2(1.32mg, 1.37. mu. mol, 1.55% yield). MS (ESI) M/z 851[ M +1]]⁺，¹H NMR(400MHz,CD₃OD)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 procedure of example 1, example 3 was obtained by reductive amination starting from intermediate compounds Z-28 and Z-16. MS (ESI) M/z 851[ M +1]]⁺，¹H NMR(400MHz,DMSO-d₆)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

Referring to the synthesis method of example 1, intermediate Z-14 and intermediate Z-28 are used as raw materials, and example 4 is obtained through reductive amination. MS (ESI) M/z 837[ M +1]]⁺，¹H NMR(400MHz,DMSO-d₆)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

Example 5 was obtained by a condensation reaction using intermediates Z-26 and Z-28 as raw materials and HATU as a condensing agent, according to the synthesis method of example 2. MS (ESI) M/z 855[ M +1]]⁺,¹H 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

By reference to the synthesis of example 1, starting from Z-1 and Z-28, reductive amination gave example 6, ms (esi) M/Z841 [ M +1]]⁺，¹H 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, while replacing Z-2 in step with the corresponding SM-aldehyde in the table. The same procedure was followed, with the substitution of SM-amine for Z-28 in step (a), to give the following examples 7 to 27:

example 27 Synthesis of Compound 27

Step 1, Synthesis of intermediate 27-1

To a solution of intermediate Z-35(0.9g,2.66mmol) in DMF (10mL) was added 4-pentyn-1-ol (448mg,5.32mmol), TEA (806mg,7.98mmol), CuI (101mg,0.5mmol) and Pd (PPh)₃)₂Cl₂(187mg,0.27mmol), the mixture was purged with nitrogen several times, and then N₂The temperature is raised to 90 ℃ under the protection of (1), and the reaction is stirred at the temperature again for 12 hours. After 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 isolated and purified (100% EA) with a silica gel column to give 27-1(190mg,0.56mmol, 21%) ms (esi) M/z ═ 342[ M +1] yield]⁺。

Step 2, Synthesis of intermediate 27-2

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

Step 3, Synthesis of Compound 27

To intermediate Z-28(70mg,0.13mmol) in DCM (7mL) was added 27-2(58.4mg,0.17mmol) and a catalytic amount of acetic acid, the mixture was stirred at room temperature for 1h, then NaBH (OAc) was added₃(84mg,0.397mmol) and the reaction was stirred at room temperature for an additional 4 h. After completion of the reaction, the reaction solution was concentrated, the crude product was diluted with water, extracted with ethyl acetate, the combined organic phases were washed with water and saturated brine, dried over anhydrous sodium sulfate, and the concentrated crude product was isolated and purified by means of mHPLC to give compound 27(45mg,0.05mmol, yield 40%). MS (ESI) M/z 852[ M + H ]]⁺。

Example 28 Synthesis of Compound 28

Referring to the procedure from step 1 to step 3 of the synthesis of example 27, compound 28 was obtained by substituting intermediate Z-33 for intermediate Z-35 in step 1, without changing the remaining procedures. MS (ESI) M/z 852[ M + H ]]⁺。

Example 29 Synthesis of Compound 29

Referring to the procedure from step 1 to step 3 of the synthesis of example 27, compound 29 was obtained by substituting intermediate Z-34 for intermediate Z-35 in step 1, without changing the remaining procedures. MS (ESI) M/z 852[ M + H ]]⁺。

Examples 30 to 38, Synthesis of Compounds 30 to 38

Reference is made to the procedure for the reductive amination of step 3 of the synthesis of example 27, while replacing 27-2 of step with the corresponding SM-aldehyde in the table. The same procedure was followed, with the substitution of SM-amine for Z-30 in step (a), to give the following examples 30 to 38:

example 39 Synthesis of Compound 39

Step 1, Synthesis of intermediate 39-1

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

Step 2, Synthesis of intermediate 39-2

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

Step 3, Synthesis of Compound 39

Referring to the procedure of example 27, step 3, intermediate 39-2 was used in place of 27-2 and the remaining procedures were identical (i.e., in NaBH (AcO))₃As a reagent, DCM plus a catalytic amount of acetic acid as a solvent, reductive amination) to give compound 39. MS (ESI) M/z 856[ M +1]]⁺。

Example 40 Synthesis of Compound 40

Referring to the procedure from step 1 to step 3 of the synthesis of example 39, intermediate 40-2 was used in place of intermediate 39-2 in step 1, and the remaining procedures were not changed to give compound 28. MS (ESI) M/z 856[ M + H ]]⁺。

Example 41 Synthesis of Compound 41

Referring to the procedure from step 1 to step 3 of the synthesis of example 39, intermediate 29-1 was used in place of intermediate 27-1 in step 1, and the remaining procedures were not changed 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 for the reductive amination of step 3 of the synthesis of example 27, while replacing 27-2 of step with the corresponding SM-aldehyde in the table. The same procedure was followed, with the substitution of SM-amine for Z-30 in step (a), to give the following examples 42 to 50:

example 51 Synthesis of Compound 51Z-28

Step 1, Synthesis of intermediate 51-1

Adding TFA dropwise into a mixture of Z-28(42.58mg, 80.54. mu. mol) and Z-40(42.58mg, 80.54. mu. mol) in DCM (2mL), adjusting pH to 5-6, stirring for 30min, and adding NaBH (OAc)₃(51.21mg, 241.63. mu. mol), and 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, and 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(12mg, 11.89. mu. mol, 14.76% yield) as a yellow solid.

Step 2, Synthesis of Compound 51

To a solution of 51-1(12mg, 11.89. mu. mol) in DCM (2mL) was added TFA (1.36mg, 11.89. mu. mol,2mL) at room temperature, and the mixture was stirred at room temperature for 1And (4) hours. After completion of the reaction, the reaction was concentrated, and the resulting crude product was redissolved in DCM (2mL), TEA (725.50mg,7.17mmol,1mL) was added at room temperature, and the reaction was stirred for 30 minutes. The mixture was concentrated and the crude product was purified by pre-HPLC to give 51(1.14mg,1.14umol, 9.56% yield, 99% purity, TF) as a yellow solid. MS (ESI) M/z 879[ M + H ]]⁺. Nuclear magnetic spectrum:¹H 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

Referring to the synthesis steps 1 and 2 of compound 51, compound 52 can be obtained by using intermediate Z-30 instead of Z-28 in step 1 as a starting material and performing the same operation. MS (ESI) M/z 879[ M + H ]]⁺. Nuclear magnetic spectrum:¹H NMR(400MHz,DMSO-d₆)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

By reductive amination of Z-28 and Z-36 starting from the synthesis of step 1 of example 51, compound 53 can be obtained, MS (ESI) m/Z ═879[M+H]⁺. Nuclear magnetic spectrum:¹H NMR(400MHz,DMSO-d₆)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

Referring to the procedure of example 51, step 1, starting from SM-aldehyde and SM-amine in the following table, the compounds of the corresponding examples in the table can be obtained by reductive amination.

Example 63 Synthesis of Compound 63

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

Reaction ofAfter completion, the reaction mixture was filtered, the filtrate was concentrated and purified by pre-HPLC to give compound 63(9.38mg,0.009mmol, 16% yield, 99.4% purity, TF) as a yellow solid, ms (esi) M/z 893[ M + H ═]⁺。

1H NMR(400MHz,Methanol-d₄)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).

EXAMPLE 64 TO EXAMPLE 74 Synthesis of Compounds 64 to 74

By following the procedure of example 63, the compounds of the corresponding examples in the table can be obtained by condensation reaction under the same conditions using SM-amine and SM-acid as starting materials in the table below.

Example 75 Synthesis of Compound 75

To a solution of Z-30(50mg, 94.58. mu. mol) in DCM (4mL) was added Z-42(31.24mg, 94.58. mu. mol), and the reaction was carried outThe mixture was stirred at room temperature for 30min, and then AcOH was added dropwise to adjust the pH of the reaction solution to 3 to 4. Add NaBH (AcO)₃(60.14mg, 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, and the combined organic phases were washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated to give a crude product, which was separated and purified by pre.hplc to give compound 75(33.8mg,35.32 μmol, 37.34% yield, TF), ms (esi) M/z-843 [ M + H ═ 843]+. Nuclear magnetic spectrum:¹H 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

Referring to the procedure of example 75, starting from SM-amine and Z-42 in the following Table, the compounds of the corresponding examples in the Table can be obtained by reductive amination under the same conditions.

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 table can be obtained by reductive amination under the same conditions using the intermediate Z-28 as a raw material and aldehydes having the corresponding structures in the table below as raw materials.

Example 82 Synthesis of Compound 82

Referring to the procedure of example 63, intermediate Z-28 and SM-acid in the following Table were subjected to condensation reaction under the same conditions to obtain compound 82 of example.

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 table can be obtained by reductive amination under the same conditions using the intermediate Z-30 as a raw material and aldehydes having the corresponding structures in the table below as raw materials, respectively.

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 table can be obtained by reductive amination under the same conditions using the intermediate Z-31 as a raw material and aldehydes having the corresponding structures in the table below as raw materials.

Example 91 Synthesis of Compound 91

Referring to the procedure of example 63, intermediate Z-31 and SM-acid in the following Table were subjected to condensation reaction under the same conditions to give example compound 91.

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 table can be obtained by reductive amination under the same conditions using the intermediate Z-32 as a raw material and aldehydes having the corresponding structures in the table below as a raw material, respectively.

Example 94 Synthesis of Compound 94

Step 1, Synthesis of intermediate 94-a

To Z-31(36mg,67.83 μmol) in DCM (2mL) was added TEA to pH 9-10, followed by Z-43(41.89mg,81.40 μmol), stirred at rt for 30min, then adjusted to pH 4-5 with acetic acid. After stirring for 5 minutes, NaBH (OAc) was added in one portion₃(43.13mg, 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 and extraction was performed, the organic phase was washed with water, brine, dried, concentrated and the crude product was isolated and purified by Pre-TLC (DCM/MeOH ═ 10/1) to give intermediate 94-a (69mg,67.03 μmol, 98.82% yield). MS (ESI) M/z 1029[ M +1]]⁺。

Step 2, Synthesis of Compound 94

Adding TFA (18.04mg,158.25 mu mol) into a DCM (2mL) solution of 94-a (70mg,68.00 mu mol) at room temperature, stirring the mixed solution at room temperature for 2 hours, concentrating the reaction solution after the reaction is finished, and separating and purifying a crude product by pre-HPLC to obtain 94; (30.88mg, 28.96. mu. mol, 42.58% yield, 95% purity, TF) yellow solid. MS (ESI) M/z 899[ M + H ]]⁺. Nuclear magnetic spectrum: 1H NMR (400MHz, Methanol-d4) δ 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 ═ 2.1Hz,1H),7.05(d, J ═ 8.7Hz,1H), 5.20-5.12 (m,1H), 4.56-4.44 (m,3H), 4.35-4.26 (m,1H), 4.16-4.08 (m,1H), 3.95-3.86 (m,2H),3.84(s,3H), 3.79-3.67 (m,2H),3.57(t, J ═ 5.6, 2H),3.46(t, 6J ═ 3.18, 3.80 (m,2H), 3.7-4.80 (m,2H), 3.31H), 3.49-2H, 1H), 3.49 (m,2H), 3.7-4.7H), 3.7 (m,2H), 3.9H), 3.31H, 3.9 (m,2H), 3.9H, 2H), 3.9H, 3.9 (m, 3.9H), 3.9 (m,2H), 3.9H), 2H), 3.9H, 2H), 3.9 (m,2H), 3.9 (m,2H), 3.9H), 2H), 2H, 1H, 2H), 3.9 (m,2H), 2H, 1H, 2H, 1H, 2H, 1H, 2H, 1H), 3.9H, 2H, 1H, 2H, 1H),3.9 (m,1H, 2H, 1H, 2H, 1H, 2H, 1H, 2H, 1H, 2H, 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

Referring to the procedure of example 94, step 1 to step 2, starting from SM-aldehyde and SM-amine in the following table, the compounds of the corresponding examples in the table were obtained by reductive amination.

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

Test example 1 EGFR kinase Activity detection

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

KinEASE^TMIn the kit (CISBIO), the specific operation is as follows: adding 125pM EGFR (T790M/C797S/L858R) kinase protein, 300nM TK Substrate-biotin, 0.8. mu.M ATP into a 384-well plate, reacting in a total volume of 10. mu.L, mixing uniformly, and incubating at room temperature for 40 min; 37.5nM Streptavidin-XL665 and 5. mu.L/well TK Antibody-Cryptate were added, the reaction volume was 20. mu.L, the mixture was mixed and incubated at room temperature for 60 minutes, the fluorescence intensities of the excitation wavelength 320nM and the emission wavelength 620nM and 665nM were measured with a microplate reader, and the FRET signal intensity (665nM fluorescence intensity/620 nM fluorescence intensity 10000) was calculated. When the inhibition of the compound on the kinase activity is detected, the compound and the kinase protein are pre-incubated for 15-60 minutes at room temperature, and then a substrate and ATP are added for reaction. The activity data are as follows:

test example 2 detection of formation of ternary Complex of EGFR-PROTAC-CRBN/DDB1

A series of concentrations of the compound of the example and EGFR (T790M/C797S/L858R) recombinant protein and CRBN/DDB1 recombinant protein complex were added to a 384-well plate, the reaction buffer was 1 XPBS pH 7.4, incubated at room temperature for 120 minutes, further added with 5nM Anti GST-Eu cryptate and 6.67nM, and Ant 6His-XL665, mixed well and incubated at room temperature for 60 minutes, the fluorescence intensity at excitation wavelength 320nM emission wavelength 620nM and 665nM was measured with a microplate reader, and the FRET signal intensity (665nM fluorescence intensity/620 nM fluorescence intensity 10000) was calculated. Experimental results as shown in fig. 1, the compounds of the present invention are capable of forming ternary complexes.

Test example 3 detection of EGFR degradation at cellular level

Coating a 96-hole blackboard with a transparent bottom with collagen and fibronectin, and connectingThe strain Ba/F3-EGFR (T790M/C797S/L858R) overexpresses cells (2 x 10^4 cells per well) and 5% CO at 37 deg.C₂The culture was carried out overnight in an incubator. The following day, the medium was removed from the cells, 150. mu.L of fresh medium was added, 50. mu.L of medium containing 4-fold final concentration of the compound of the example was added, and the mixture was placed in an incubator at 37 ℃ and 5% CO₂The culture was continued under the conditions for 16 hours. Thereafter, 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 15 min. After cell fixation, 150. mu.L of 0.1% (v/v) Triton X-100 permeabilized cells were used and incubated at room temperature for 15 min. Cells were blocked with 1% blocking reagent (Roche, cat # 11096176001) at 100. mu.L per well and incubated for 60min at room temperature. After blocking, 50. mu.L of EGFR antibody (CST, cat. No.: 4267S, dilution ratio 1:250) diluted with 1% (w/v) blocking reagent was added to each well and incubated at room temperature for 2 hours. EGFR antibody was removed from the wells and cells were washed with 0.05% (v/v) PBST, 200. mu.L per well, in 4 replicates. Then, 1% (w/v) of goat anti-rabbit fluorescent secondary antibody (Invitrogen, cat # A32731) diluted with blocking reagent and DAPI dye solution (Thermofisiher, cat # 62247) (fluorescent secondary antibody dilution ratio 1:250, DAPI dilution ratio 1:2000) were added thereto, 50. mu.L per well, and incubated at 300rpm on a shaker at room temperature for 2 hours in the dark. The wells were then removed of antibody and the cells were washed with 0.05% (v/v) PBST, 200. mu.L per well, in 4 replicates (the wash in the wells was retained for the last time). 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 0.05 s. The data were analyzed using In cell analyzer work software to obtain the intracellular EGFR fluorescence signal value and the background value around the cells. The obtained data were subsequently processed using Excel by first subtracting the background value around the cells from the intracellular EGFR fluorescence signal value to obtain a normalized intracellular EGFR fluorescence signal value. Next, the relative level of EGFR after compound treatment relative to DMSO control (100% based on DMSO treatment) was calculated, analyzed using GraphPad Prism software, and DCs were obtained₅₀Values and fitted curves.

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

Claims

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

wherein,

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

y is selected from N or CR^Y1；

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

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

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

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

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

ring B is selected from

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

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

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

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

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

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

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

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

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

L^A3is selected from

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

Or none.

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

R⁴、R⁵each independently selected from hydrogen.

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

wherein,

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

y is selected from N or CR^Y1；

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

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

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

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

wherein,

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

y is selected from N or CR^Y1；

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

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

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

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

wherein,

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

y is selected from N or CR^Y1；

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

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

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

l is- (L)^A)_q-；

q is an integer greater than or equal to 1;

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

l is selected from the following groups:

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

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

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

R⁴、R⁵each independently selected from hydrogen;

w is selected from C (O), CH₂、NH；

R^BIs selected from methyl;

l is selected from the following linking groups:

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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