WO2024188281A1 - Compound having quinazoline structure and use thereof - Google Patents

Abstract

Disclosed in the present invention are a compound having a quinazoline structure and the use thereof. Specifically, the present invention relates to a compound as represented by formula (VI), a stereoisomer thereof and a pharmaceutically acceptable salt thereof.

Description

Compounds containing quinazoline structure and their application

This application claims the following priority:

CN202310245836.7, March 13, 2023;

CN202310244093.1, March 14, 2023;

CN202310286647.4, March 22, 2023;

CN202310873122.0, July 14, 2023;

CN202310918759.7, July 24, 2023;

CN202311060914.2, August 21, 2023;

CN202311265591.0, September 27, 2023;

CN202311677606.4, December 06, 2023;

CN202410116075.X, January 26, 2024.

Technical Field

The present invention relates to a class of compounds containing quinazoline structure and applications thereof, and in particular to a compound represented by formula (VI), a stereoisomer thereof and a pharmaceutically acceptable salt thereof.

Background Art

RAS is a functionally important guanine nucleoside binding protein, and mutated RAS proteins are considered to be important carcinogenic factors. RAS mutations are found in 20% of human tumors, of which 85% are KRAS. KRAS protein has 188 amino acids, and the most common mutations occur at positions 12, 13, and 61, especially at position 12, where the most common mutations are G12D, G12V, and G12C.

Why does mutated KRAS cause cancer? This is because the conversion of KRAS between inactive and activated states is regulated by two types of factors: guanine nucleotide exchange factors (GEFs) that convert KRAS from inactive to activated states; and GTPase activating proteins (GAPs) that convert KRAS from activated to inactive states. When KRAS mutates, its ability to bind to GAPs is greatly reduced, thereby keeping the KRAS protein in an activated state, leading to the occurrence of cancer.

In recent years, KRAS protein inhibitors with G12C mutations have made great breakthroughs. Amgen's Sotorasib has achieved good results in clinical practice and has been approved by the FDA. However, there has been no significant progress in other KRAS mutations. This is mainly because the KRAS protein is small and has weak binding to small molecules. In addition to covalent bonding, it is difficult to find other alternatives.

Proteolysis-Targeting Chimeras (PROTACs) are hybrid bifunctional small molecule compounds, one end of which is the part that binds to the target, and the other end is the E3 ligand, and the two are connected by a linker fragment. PROTAC is a new class of drugs with good prospects. When one end binds to the target, the E3 ligand part at the other end induces ubiquitination, thereby degrading the target protein. The biggest advantage of PROTAC technology is that it can make some targets that are considered undruggable become druggable - there is no need for particularly strong binding ability, as long as it can induce ubiquitination, the function of the target protein can be inhibited.

The application of PROTAC technology to the very important but difficult-to-drug KRAS mutant protein is a very promising strategy. It is expected to overcome the drug resistance problem of KRAS mutant protein, so that these patients can have drugs available.

Summary of the invention

The present invention provides a compound of formula (I), a stereoisomer thereof and a pharmaceutically acceptable salt thereof,

in,

Ring A is selected from 8-16 membered heterocycloalkyl, 8-16 membered heterocycloalkenyl and 8-16 membered heteroaryl, wherein the 8-16 membered heterocycloalkyl, 8-16 membered heterocycloalkenyl and 8-16 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R _c ;

Each R _c is independently selected from H, F, Cl, Br, I, CN, C _1-6 alkyl, C 1-6 alkoxy, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, and the C _1-6 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl and _3-6 membered heterocycloalkyl are independently optionally substituted by 1, 2 or 3 R _o ;

G ₁ is selected from phenyl and 5-6 membered heteroaryl, and the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R _a1 ;

each R _a1 is independently selected from H, F, Cl, Br, I, C _1-3 alkyl, C _{3-6 cycloalkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl, and the C 1-3 alkyl} , C _3-6 cycloalkyl, 4-6 membered heterocycloalkyl and _5-6 membered heteroaryl are independently optionally substituted by 1, 2 or 3 R _o ;

_G2 is absent or selected from phenyl, naphthyl, pyridyl, benzothiazolyl and indazolyl, wherein the phenyl, naphthyl, pyridyl, benzothiazolyl and indazolyl are independently optionally substituted by 1, 2 or 3 _Rb ;

Each R _b is independently selected from D, F, Cl, Br, I, OH, NH ₂ , C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy and C _1-6 alkylamino;

G ₃ is absent or is selected from C _1-20 alkyl, C _3-20 cycloalkyl, 3-20 membered heterocycloalkyl and NR ¹ R ² , wherein the C _1-20 alkyl, C _3-20 cycloalkyl and 3-20 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R ₀ ;

R ¹ is selected from C _1-6 alkyl, wherein the C _1-6 alkyl is optionally substituted by 1, 2 or 3 R ¹¹ ;

R ² is selected from 4-6 membered heterocycloalkyl, which is optionally substituted by 1, 2 or 3 R ²¹ ;

each R ¹¹ and R ²¹ is independently selected from F, Cl, Br, I, CN, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ , CH ₂ F and OCH ₃ ;

L ₁ is selected from bicyclo[2.2.2]octyl, 2,3-dihydro-1H-indenyl, phenyl and pyridinyl, wherein the bicyclo[2.2.2]octyl, 2,3-dihydro-1H-indenyl, phenyl and pyridinyl are each independently optionally substituted with 1, 2 or 3 R ₀ ;

_L2 is selected from -L _a -L _b -L _c -L _d -;

Each of _La , _Lb , _Lc and _Ld is independently selected from a single bond, -O-, -NH-, -N( _C1-3 alkyl)-, pyrrolidinyl, piperidinyl, piperazinyl, _C1-3 alkyl and -(C=O)-;

_L3 is selected from -NH- and 5-6 membered heteroaryl, wherein the -NH- and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 _R0 ;

R ₁₀ is selected from C _1-6 alkyl, C _3-6 cycloalkyl and 4-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _3-6 cycloalkyl and 4-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R ₀ ;

R ₂₀ is selected from H and OH;

R ₃₀ and R ₄₀ are independently selected from H and C _1-6 alkyl, and the C _1-6 alkyl is optionally substituted by 1, 2 or 3 R ₀ ;

Alternatively, R ₃₀ , R ₄₀ and the carbon atom to which they are connected together form a C _3-6 cycloalkyl group or a 4-6 membered heterocycloalkyl group, and the C _3-6 cycloalkyl group and the 4-6 membered heterocycloalkyl group are each independently optionally substituted by 1, 2 or 3 R ₀ ;

R ₅ is selected from -L ₄ -R ₄ ;

L ₄ is selected from a single bond, -CH ₂ -, -CD ₂ -, -CH(C _1-3 alkyl)-, -O-, -S-, -NH- and -N(C _1-3 alkyl)-;

_R4 is selected from _C1-6 alkyl, 4-6 membered heterocycloalkyl, 5-6 membered heteroaryl and _C1-6 alkyl-N( _C1-3 alkyl)( _OC1-3 alkyl), wherein the _C1-3 alkyl, _C1-6 alkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R _{e1 ;}

Each R ₆ and R ₇ is independently selected from H, F, Cl, Br, I and C _1-3 alkyl, wherein the C _1-3 alkyl is optionally substituted with 1, 2 or 3 R ₀ ;

each R ₈ and R ₉ is independently selected from H, D, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ and CH ₂ F;

Each R _e1 is independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ , CH ₂ F and OCH ₃ ;

Each R ₀ is independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ , CH ₂ F and OCH ₃ .

The present invention provides a compound of formula (I), a stereoisomer thereof and a pharmaceutically acceptable salt thereof,

in,

Ring A is selected from 8-16 membered heterocycloalkyl, 8-16 membered heterocycloalkenyl and 8-16 membered heteroaryl, wherein the 8-16 membered heterocycloalkyl, 8-16 membered heterocycloalkenyl and 8-16 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R _c ;

Each R _c is independently selected from H, F, Cl, Br, I, CN, C _1-6 alkyl, C 1-6 alkoxy, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, and the C _1-6 alkyl, C _1-6 alkoxy, C _3-6 cycloalkyl and _3-6 membered heterocycloalkyl are independently optionally substituted by 1, 2 or 3 R _o ;

G ₁ is selected from phenyl and 5-6 membered heteroaryl, and the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R _a1 ;

each R _a1 is independently selected from H, F, Cl, Br, I, C _1-3 alkyl, C _{3-6 cycloalkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl, and the C 1-3 alkyl} , C _3-6 cycloalkyl, 4-6 membered heterocycloalkyl and _5-6 membered heteroaryl are independently optionally substituted by 1, 2 or 3 R _o ;

_G2 is absent or selected from phenyl, naphthyl, pyridyl, benzothiazolyl and indazolyl, wherein the phenyl, naphthyl, pyridyl, benzothiazolyl and indazolyl are independently optionally substituted by 1, 2 or 3 _Rb ;

Each R _b is independently selected from D, F, Cl, Br, I, OH, NH ₂ , C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy and C _1-6 alkylamino;

_G3 is absent or selected from _C1-20 alkyl, _C3-20 cycloalkyl and 3-20 membered heterocycloalkyl, wherein the _C1-20 alkyl, _C3-20 cycloalkyl and 3-20 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 _R0 ;

L ₁ is selected from bicyclo[2.2.2]octyl, 2,3-dihydro-1H-indenyl, phenyl and pyridinyl, wherein the bicyclo[2.2.2]octyl, 2,3-dihydro-1H-indenyl, phenyl and pyridinyl are each independently optionally substituted with 1, 2 or 3 R ₀ ;

_L2 is selected from -L _a -L _b -L _c -L _d -;

Each of _La , _Lb , _Lc and _Ld is independently selected from a single bond, -O-, -NH-, -N( _C1-3 alkyl)-, pyrrolidinyl, piperidinyl, piperazinyl, _C1-3 alkyl and -(C=O)-;

_L3 is selected from -NH- and 5-6 membered heteroaryl, wherein the -NH- and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 _R0 ;

R ₁₀ is selected from C _1-6 alkyl, C _3-6 cycloalkyl and 4-6 membered heterocycloalkyl, wherein the C _1-6 alkyl, C _3-6 cycloalkyl and 4-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R;

R ₂₀ is selected from H and OH;

R ₃₀ and R ₄₀ are independently selected from H and C _1-6 alkyl, and the C _1-6 alkyl is optionally substituted by 1, 2 or 3 R ₀ ;

Alternatively, R ₃₀ , R ₄₀ and the carbon atom to which they are connected together form a C _3-6 cycloalkyl group or a 4-6 membered heterocycloalkyl group, and the C _3-6 cycloalkyl group and the 4-6 membered heterocycloalkyl group are each independently optionally substituted by 1, 2 or 3 R ₀ ;

R ₅ is selected from -L ₄ -R ₄ ;

L ₄ is selected from a single bond, -CH ₂ -, -CD ₂ -, -CH(C _1-3 alkyl)-, -O-, -S-, -NH- and -N(C _1-3 alkyl)-;

_R4 is selected from _C1-6 alkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl, wherein the _C1-6 alkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R _e1 ;

Each R ₆ and R ₇ is independently selected from H, F, Cl, Br, I and C _1-3 alkyl, wherein the C _1-3 alkyl is optionally substituted with 1, 2 or 3 R ₀ ;

each R ₈ and R ₉ is independently selected from H, D, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ and CH ₂ F;

Each R _e1 is independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ , CH ₂ F and OCH ₃ ;

Each R ₀ is independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ , CH ₂ F and OCH ₃ .

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:
Other variables are as defined in the present invention.

The present invention provides a compound of formula (IV), a stereoisomer thereof and a pharmaceutically acceptable salt thereof.

in,

_G3 is selected from 7-8 membered heterocycloalkyl and The 7-8 membered heterocycloalkyl group is optionally substituted by 1, 2 or 3 R ₀ , respectively, independently;

R ₂₀ is selected from H and OH;

R ₅ is selected from -O-4-6 membered heterocycloalkyl and -OC _1-3 alkyl-N(C _1-3 alkyl)(OC _1-3 alkyl), wherein the C _1-3 alkyl and the 4-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _e1 ;

each R ₆ and R ₇ is independently selected from H, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ and CH ₂ F;

each R ₈ and R ₉ is independently selected from H, D, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ and CH ₂ F;

each R _b is independently selected from D, F, Cl, Br, I, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ and CH ₂ F;

Each R _e1 is independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ , CH ₂ F and OCH ₃ ;

Each R ₀ is independently selected from H, F, Cl, Br, I, CN, OH, NH ₂ , CH ₃ , CF ₃ , CHF ₂ , CH ₂ F and OCH ₃ .

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

wherein R ₅ , R ₆ , R ₇ and R _b are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

wherein R ₅ , R ₆ , R ₇ and R _b are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

wherein R ₅ , R ₆ , R ₇ and R _b are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned _G2 is selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the above R _b is selected from F and CH ₃ , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₅ is selected from -OR ₄ , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₅ is selected from -O-4-6 membered heterocycloalkyl and -OC _1-3 alkyl-N(C _1-3 alkyl)(OC _1-3 alkyl), and the C _1-3 alkyl and 4-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _e1 , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₅ is selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₅ is selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the above L ₄ is selected from -O-, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned _G3 is selected from 7-8 membered heterocycloalkyl and The 7-8 membered heterocycloalkyl group is optionally substituted by 1, 2 or 3 R _0, respectively, and other variables are as defined herein.

In some embodiments of the present invention, the above G ₃ is selected from 7-8 membered heterocycloalkyl, and the 7-8 membered heterocycloalkyl is optionally substituted by 1, 2 or 3 R, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned _G3 is selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned _G3 is selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned _G3 is selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the above G ₁ is selected from phenyl, and the phenyl is optionally substituted by 1, 2 or 3 R _a1 , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above G ₁ is selected from phenyl, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₁₀ is selected from isopropyl, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₂₀ is selected from OH, and other variables are as defined in the present invention.

In some embodiments of the present invention, R ₃₀ and R ₄₀ are independently selected from H and -CH ₂ OH, and other variables are as defined in the present invention.

In some embodiments of the present invention, R ₃₀ is selected from H, R ₄₀ is selected from -CH ₂ OH, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₆ is selected from CH ₃ , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₇ is selected from H, and other variables are as defined in the present invention.

The present invention provides a compound of formula (VI), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

in,

T is selected from S and Se;

R ₁ is selected from H, D and C _1-3 alkyl;

_R2 is selected from H, D and _C1-3 alkyl;

R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl;

R ₅ is selected from -OR ₄ ;

R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _a ;

each R ₆ and R ₇ is independently selected from H, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ and CH ₂ F;

Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl )-( _OC1-3alkyl ), wherein said -C(=O) _C1-3alkyl , -N( _C1-3alkyl )-( _OC1-3alkyl ) are independently optionally substituted by 1, 2 or 3 R;

Each R _b is independently selected from D, F, Cl, Br, I, OH, NH ₂ and CH ₃ ;

Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH;

m is selected from 0, 1, 2, 3 and 4;

Provided that: when T is selected from S, _R1 and _R2 are both H, _R3 is not H and F, and m is not 0.

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

in,

R ₁ is selected from H, D and C _1-3 alkyl;

_R2 is selected from H, D and _C1-3 alkyl;

R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl;

R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _a ;

R ₅ is selected from -O-4-6 membered heterocycloalkyl and -OC _1-3 alkyl-N(C _1-3 alkyl)(OC _1-3 alkyl), wherein the C _1-3 alkyl is each independently optionally substituted by 1, 2 or 3 R;

Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl , wherein said -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl are independently optionally substituted by 1, 2 or 3 R;

Each R _b is independently selected from D, F, Cl, Br, I, OH, NH ₂ and CH ₃ ;

Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH;

m is selected from 0, 1, 2, 3 and 4;

Provided that: when _R1 and _R2 are both H, _R3 is not H and F, and m is not 0.

The present invention provides a compound of formula (II), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

in,

R ₁ is selected from H, D and C _1-3 alkyl;

_R2 is selected from H, D and _C1-3 alkyl;

R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl;

R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _a ;

Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl , wherein said -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl are independently optionally substituted by 1, 2 or 3 R;

Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH;

m is selected from 0, 1, 2, 3 and 4;

Provided that: when _R1 and _R2 are both H, _R3 is not H and F, and m is not 0.

The present invention provides a compound of formula (II), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

in,

R ₁ is selected from H, D and C _1-3 alkyl;

_R2 is selected from H, D and _C1-3 alkyl;

R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl;

R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _a ;

Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl , wherein said -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl are independently optionally substituted by 1, 2 or 3 R;

Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH;

m is selected from 0, 1, 2, 3 and 4;

Provided that: when _R1 and _R2 are both H, _R3 is not H and m is not 0.

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

in,

R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 R _a ;

R ₅ is selected from -O-4-6 membered heterocycloalkyl and -OC _1-3 alkyl-N(C _1-3 alkyl)(OC _1-3 alkyl);

R ₃ , _Ra and m are as defined herein.

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

wherein R ₁ , R ₂ , R ₃ , R ₄ and m are as defined in the present invention.

In some embodiments of the present invention, the above compound has a structure shown in formula (II-1):

in,

R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 R _a ;

R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl;

Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl , wherein said -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl are independently optionally substituted by 1, 2 or 3 R;

Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH;

m is selected from 0, 1 and 2.

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

in,

R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 R _a ;

R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl;

Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl , wherein said -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl are independently optionally substituted by 1, 2 or 3 R;

Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH;

m is selected from 0, 1 and 2.

In some embodiments of the present invention, the above R ₁ is selected from H and D, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₂ is selected from H, D and CH ₃ , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₃ is selected from H and Cl, and other variables are as defined in the present invention.

In some embodiments of the present invention, each _Ra is independently selected from -C(=O)CH ₃ and -N(CH ₃ )-OCH ₃ , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₄ is selected from propyl, isopropyl, tetrahydropyranyl and pyrrolidinyl, and the propyl, isopropyl, tetrahydropyranyl and pyrrolidinyl are independently and optionally substituted by 1, 2 or 3 _Ra , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₄ is selected from Said are each independently optionally substituted by 1, 2 or 3 _Ra , and other variables are as defined herein.

In some embodiments of the present invention, the above R ₄ is selected from Other variables are as defined in the present invention.

In some embodiments of the present invention, the structural unit Selected from Other variables are as defined in the present invention.

Some other solutions of the present invention are obtained by arbitrarily combining the above variables.

In some embodiments of the present invention, the above-mentioned compound, its stereoisomers and pharmaceutically acceptable salts thereof are selected from:

wherein R ₃ , R ₄ and m are as defined in the present invention.

The present invention provides the following compounds, their stereoisomers and pharmaceutically acceptable salts:

In some embodiments of the present invention, the above compound is selected from:

In some embodiments of the present invention, the above compound is selected from:

The present invention also provides the use of the above compound, its stereoisomer or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating a disease associated with a KRAS mutant protein.

In some embodiments of the present invention, the above-mentioned diseases associated with KRAS mutant protein are selected from solid tumors with KRAS G12D mutation.

In some embodiments of the present invention, the solid tumor with the above-mentioned KRAS G12D mutation is selected from pancreatic cancer, lung cancer and colon cancer.

The compounds of the present invention are synthesized by the following route:

Synthetic route 1:

Synthetic route 2:

The present invention also refers to the following test method:

Test method 1: In vitro cell proliferation assay

Experimental Materials:

RPMI-1640 medium was purchased from GIbco, penicillin/streptomycin antibiotics were purchased from Vicente, and fetal bovine serum was purchased from Biosera. 3D CellTiter-Glo (cell viability chemiluminescence detection reagent) reagent was purchased from Promega. AsPC-1/A375 and other cell lines were purchased from ATCC, and Envision multi-label analyzer was purchased from PerkinElmer.

Experimental methods:

The cells were seeded in an ultra-low attachment 96-well U-shaped plate, with 80 μL of cell suspension per well, containing 1000 selected cells. The cell plate was placed in a carbon dioxide incubator for overnight culture.

The compound to be tested was diluted 5-fold into 8 concentrations using a dispenser, i.e., from 2mM to 25.6nM, and a double-well experiment was set up. 78μL of culture medium was added to the middle plate, and then 2μL of the gradient diluted compound per well was transferred to the middle plate according to the corresponding position. After mixing, 20μL of each well was transferred to the cell plate. The concentration range of the compound transferred to the cell plate was 10μM to 0.128nM. The cell plate was placed in a carbon dioxide incubator and cultured for 6 days. Prepare another cell plate and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis.

Add 100 μL of cell viability chemiluminescent detection reagent to the cell plate and incubate at room temperature for 30 min to stabilize the luminescent signal. Read using a multi-label analyzer.

Data Analysis:

The raw data were converted into inhibition rate using the equation (Sample-Min)/(Max-Min)*100%, and the _IC50 value was obtained by four-parameter curve fitting (obtained using the "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism).

Test method 2: Cellular KRAS G12D protein degradation experiment

Experimental Materials:

RPMI-1640 medium was purchased from GIbco, penicillin/streptomycin antibiotics were purchased from Vicente, and fetal bovine serum was purchased from Biosera. Protein primary and secondary antibodies were purchased from Cell Signaling Technology. AsPC-1/A375 and other cell lines were purchased from ATCC, and 4% paraformaldehyde and Triton-100 were purchased from Bio-Tech. Envision multi-label analyzer (PerkinElmer).

Experimental methods:

The cells were seeded in ultra-low attachment 96-well black-walled plates, with 80 μL of cell suspension per well, containing 100,000 cells. The cell plates were placed in a carbon dioxide incubator for overnight culture.

The compound to be tested was diluted 5-fold into 8 concentrations using a shotgun, i.e., from 2mM to 25.6nM, and a double-well experiment was set up. 78μL of culture medium was added to the middle plate, and then 2μL of the gradient diluted compound per well was transferred to the middle plate according to the corresponding position. After mixing, 20μL of each well was transferred to the cell plate. The concentration range of the compound transferred to the cell plate was 10μM to 0.128nM. After the cell plate was cultured in a carbon dioxide incubator for 24h, the cells were removed, the culture medium was discarded, and the cells were fixed with 4% PFA fixative for 30min, washed 3 times with PBS, permeabilized with 0.1% Triton for 20min, washed 3 times with PBS, blocked with 0.1% BSA solution at room temperature for 1h, incubated with protein primary antibody at 4°C overnight, washed 3 times with PBST, incubated with fluorescent secondary antibody at room temperature for 2h, washed 3 times with PBST,

Data Analysis:

Calculate the FITC/Cy5 in the sample well based on the measured fluorescence intensity

FITC/Cy5=(FITCsample-FITCblank)/(FITCsample-FITCblank)

The inhibition % under compound treatment was calculated as [1-(FITC/Cy5comp.)/(FITC/Cy5DMSO)]*100. The DC ₅₀ value can be obtained by four-parameter curve fitting (obtained by "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism).

Test method 3: AsPC-1 cell p-ERK level detection

Experimental Materials:

AsPC-1 cells were purchased from ATCC; RPMI-1640 medium was purchased from GIbco; fetal bovine serum was purchased from Hyclone; Advanced Phospho-ERK1/2 (THR202/TYR204) KIT was purchased from Bioauxilium, see Table 1 for details.

Table 1: Advanced Phospho-ERK1/2 (THR202/TYR204) KIT ingredients

Experimental methods:

1. Plant AsPC-1 cells in a 384-well cell culture plate with a white bottom, 8 μL of cell suspension per well, each well contains 7500 cells, and place the cell plate in a carbon dioxide incubator and incubate at 37 degrees overnight;

2. Dilute the compound to be tested with 100% DMSO to 3mM as the first concentration, and then use a pipette to dilute it into ten concentrations of 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1μM. Take 2μL of compound and add it to 198μL of cell starvation medium. After mixing, take 15μL of compound solution and add it to 35μL of cell starvation medium and mix. Then, add 4μL of the compound solution in the last step to the corresponding cell plate wells per well. Put the cell plate back into the carbon dioxide incubator and continue to incubate for 3h. At this time, the compound concentrations are 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1nM.

3. After the incubation, add 3 μL 5X cell lysis buffer to each well and incubate at room temperature for 30 minutes with shaking;

4. Use detection buffer to dilute Phospho-ERK1/2 Eu Cryptate antibody and Phospho-ERK1/2 d2 antibody 20 times, mix them in a 1:1 ratio, add 5 μL to each well of the cell culture plate, and incubate at room temperature for 2 hours.

5. After incubation, use a multi-label analyzer to read HTRF excitation: 320nm, emission: 615nm, 665nm.

Data Analysis:

The raw data were converted into inhibition rate using the equation (Sample-Min)/(Max-Min)*100%, and the _IC50 value was obtained by four-parameter curve fitting (log(inhibitor) vs.response--Variable slope mode in GraphPad Prism).

Max well: The reading value of the positive control well is 1X lysate

Min well: The reading value of the negative control well is 0.5% DMSO cell well cell lysate.

Technical Effects

The compound of the present invention has good anti-cell proliferation activity and KRAS degradation activity, shows excellent in vitro activity on G12D mutated cells, and has no inhibitory activity on G12D low-expressing cell A375; shows excellent G12D degradation activity, and has good inhibitory activity on downstream pERK; has good in vivo tumor inhibition effect, specifically good efficacy and safety, and can be used for the treatment of cancer, especially pancreatic cancer.

Definition and Description

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be ambiguous or unclear in the absence of a special definition, but should be understood according to the meaning commonly understood by those skilled in the art to which the invention herein belongs. When a trade name appears in this document, it is intended to refer to the corresponding commercial product or its active ingredient.

Unless otherwise specified, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit/risk ratio.

Unless otherwise specified, the term "pharmaceutically acceptable salt" refers to salts of the compounds of the invention herein, prepared from compounds with specific substituents found in the invention herein with relatively non-toxic acids or bases. When the compounds of the invention herein contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of the invention herein contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds of the invention herein contain basic and acidic functional groups and can be converted into either base or acid addition salts.

Unless otherwise specified, the pharmaceutically acceptable salts of the invention herein can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. In general, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, via the reaction of these compounds in free acid or base form with a stoichiometric amount of an appropriate base or acid to prepare.

Unless otherwise specified, the term "effective amount" or "therapeutically effective amount" refers to a non-toxic amount that can achieve the desired effect. The determination of the effective amount varies from person to person, depending on the age and general condition of the recipient, and also on the specific active substance. The appropriate effective amount in each case can be determined by a person skilled in the art based on routine experiments.

Unless otherwise specified, the compounds of the invention herein may exist in specific geometric or stereoisomeric forms. The invention herein contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the invention herein. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All of these isomers and their mixtures are included within the scope of the invention herein.

Unless otherwise specified, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

Unless otherwise specified, the terms "cis-trans isomers" or "geometric isomers" result from the inability of a ring to rotate freely about double bonds or single bonds of ring carbon atoms.

Unless otherwise specified, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

Unless otherwise indicated, the term "atropisomer (atropisomer) (or restricted configuration isomer (atropoisomer)" is a stereoisomer with a specific spatial configuration, which is caused by the restricted rotation around a single bond due to large steric hindrance. Certain compounds of the present invention may exist as atropisomers. The compounds disclosed in the present invention include all atropisomers, which may be pure individual atropisomers, or enriched in one of the atropisomers, or non-specific mixtures of each. If the rotational potential around the single bond is high enough and the mutual conversion between the conformations is slow enough, separation of isomers may be allowed. Unless otherwise specified, in the invention herein, "(+)" means dextrorotation, "(-)" means levorotation, and "(±)" means racemization.

Unless otherwise specified, in the invention herein, a solid key with a wedge shape is used. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond. and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Denotes a solid wedge bond or dotted wedge key Or use a wavy line Represents a straight solid bond or straight dashed key

Unless otherwise specified, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and can readily interconvert. If tautomerism is possible (such as in solution), chemical equilibrium of the tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some of the bonding electrons. A specific example of keto-enol tautomerism is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise specified, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

Unless otherwise specified, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

Unless otherwise specified, optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention herein is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamates are generated from amines).

Unless otherwise specified, the compounds invented herein may contain unnatural proportions of atomic isotopes on one or more atoms constituting the compound. For example, compounds may be labeled with radioactive isotopes, such as tritium ( ^3H ), iodine-125 ( ^125I ) or C-14 ( ^14C ). For another example, deuterated drugs may be formed by replacing hydrogen with heavy hydrogen. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing therapeutic effects, and extending the biological half-life of drugs. All isotopic composition changes of the compounds invented herein, whether radioactive or not, are included in the scope of the invention herein.

Unless otherwise specified, the term "optional" or "optionally" by itself or in combination with other terms means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include a variant of deuterium and hydrogen, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O) or =NR', it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or not substituted, and unless otherwise specified, the type and number of substituents may be arbitrary on the basis of chemical achievable.

Unless otherwise specified, in the present invention, when any variable (such as R) occurs more than once in the composition or structure of a compound, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 R, the group may be optionally substituted with up to two R, and each occurrence of R has independent options. In addition, combinations of substituents and/or variants thereof are permitted only if such combinations result in stable compounds.

Unless otherwise specified, in the present invention, when the number of a linking group is 0, such as -(CRR) ₀ -, it means that the linking group is a single bond.

Unless otherwise specified, in the present invention, when one of the variables is selected from a single bond, it means that the two groups connected thereto are directly connected. For example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.

Unless otherwise specified, in the present invention, when a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A.

Unless otherwise specified, in the present invention, when the listed substituents do not indicate through which atom they are connected to the substituted group, such substituents can be bonded through any atom thereof. For example, a pyridyl group as a substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

Unless otherwise specified, in the present invention, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bonds are connected, the number of H atoms at the site will decrease with the number of connected chemical bonds to become a group with a corresponding valence. The chemical bond connecting the site to other groups can be a straight solid line bond. Straight dotted key or wavy line For example, the straight solid bond in -OCH ₃ indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group. It means that any connectable site on the piperidine group can be connected to other groups through one chemical bond, including at least These four connection methods, even if the H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

When the chemical bond of a substituent intersects the chemical bond connecting two atoms on a ring, it means that the substituent can form a bond with any atom on the ring. When the atom to which a substituent is attached is not specified, the substituent can form a bond with any atom. If the atom to which the substituent is attached is in a bicyclic or tricyclic ring system, it means that the substituent can form a bond with any atom of any ring in the system. Combinations of substituents and/or variables are allowed only if the combination results in a stable compound. For example, the structural unit It means that it can be substituted at any position on the cyclohexyl group or the cyclopentyl group.

Unless otherwise specified, in the present invention, when the listed linking groups do not specify their linking direction, their linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of the reading order from left to right to form Combinations of linkers, substituents, and/or variations thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, in the present invention, the term "ring" includes a ring system containing at least one ring, each of which independently satisfies The above definition.

Unless otherwise specified, in the present invention, the number of atoms in a ring is generally defined as the number of members of the ring. For example, a "5-membered ring" or a "7-membered ring" refers to a "ring" with 5 or 7 atoms arranged around it.

Unless otherwise specified, C _nm or C _n -C _m includes any specific case of n to m (m ≥ n) carbon atoms, for example, C _1-12 includes C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ , and C ₁₂ , and also includes any range from n to n+m, for example, C _1-12 includes C _1-3 , C _1-6 , C _1-9 , C _3-6 , C _3-9 , C _3-12 , C _6-9 , C _6-12 , and C ₁₃ . _9-12 , etc.; similarly, n-membered to m-membered (i.e., nm-membered) means that the number of atoms in the ring is n to m, for example, a 3-12-membered ring includes a 3-membered ring, a 4-membered ring, a 5-membered ring, a 6-membered ring, a 7-membered ring, an 8-membered ring, a 9-membered ring, a 10-membered ring, an 11-membered ring, and a 12-membered ring, and also includes any range from n to m, for example, a 3-12-membered ring includes a 3-6-membered ring, a 3-9-membered ring, a 5-6-membered ring, a 5-7-membered ring, a 6-7-membered ring, a 6-8-membered ring, and a 6-10-membered ring, etc.

Unless otherwise specified, the term "alkyl" by itself or in combination with other terms is used to represent a straight or branched saturated hydrocarbon group consisting of a certain number of carbon atoms, which may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). In some embodiments of the present invention, examples of alkyl include, but are not limited to: methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, heptyl, octyl, etc.

In some embodiments of the present invention, _C1-20 alkyl includes but is not limited to: _C1-20 alkyl, _C1-19 alkyl, _C1-18 alkyl, _C1-17 alkyl, _C1-16 alkyl, _C1-15 alkyl, _{C1-14 alkyl, C1-13} alkyl, _C1-12 alkyl, C1-11 alkyl, _C1-10 alkyl, _C1-9 alkyl, _C1-8 alkyl, _C1-7 alkyl, C1-6 alkyl, C1-5 alkyl, C1-4 alkyl, C1-3 alkyl, C1-2 alkyl, _C2-20 alkyl, _C2-19 alkyl, C2-18 alkyl, _C2-17 alkyl, C2-16 alkyl, C2-15 _alkyl , C2-14 _alkyl , _C2-13 alkyl, _C2-12 alkyl, _C2-11 alkyl, _C1-10 alkyl, _C1-9 alkyl, _{C1-8 alkyl} , _C1-7 alkyl, _C1-6 alkyl, _C1-5 alkyl, _C1-4 alkyl, _C1-3 alkyl, _alkyl , C _3-14 alkyl, C _{3-13 alkyl, C 3-12} alkyl, C _3-11 alkyl, C 3-10 _{alkyl, C 2-9 alkyl, C 2-8 alkyl, C 2-7 alkyl, C 2-6 alkyl, C 2-5 alkyl, C 2-4 alkyl, C 2-3 alkyl, C 3-20 alkyl , C 3-19 alkyl , C 3-18 alkyl ,} C _3-17 alkyl, C 3-16 alkyl, C 3-15 alkyl, C _3-14 alkyl, C _3-13 alkyl, C _3-12 alkyl, C _3-11 alkyl, C _3-10 alkyl, C _3-9 alkyl, C _{3-8 alkyl, C 3-7 alkyl} , C _3-6 alkyl, C _3-5 alkyl, C _3-4 alkyl, C _4-20 alkyl, C 4-19 alkyl, C _4-18 alkyl, C _4-17 alkyl, C _4-16 alkyl _, C _4-15 alkyl, C _4-14 alkyl, C alkyl, C _5-20 alkyl, C _5-19 alkyl, C _5-18 alkyl, C _5-17 alkyl, C _5-16 alkyl, C 5-15 _alkyl , C _5-14 alkyl, C _5-13 alkyl, C _5-12 alkyl, C _5-11 alkyl, C _5-10 alkyl, C _5-9 alkyl, C _5-8 alkyl, C _5-7 alkyl, C _5-6 alkyl, C 6-20 alkyl, C 6-19 alkyl, C 6-18 alkyl, C _6-17 alkyl, C 6-16 alkyl, C _6-15 alkyl, C _6-14 alkyl, C _6-13 alkyl, C _5-12 alkyl, C _5-11 alkyl, C _5-10 alkyl, C _5-9 alkyl, C _5-8 alkyl, C 5-7 alkyl, C _5-6 alkyl, C 6-20 alkyl, C _6-19 alkyl, C _6-18 alkyl, C _6-17 alkyl, C _6-16 alkyl, C _6-15 alkyl, C _6-14 alkyl, C alkyl, C _7-20 alkyl, C _7-19 alkyl, C _7-18 alkyl, C _7-17 alkyl, C _7-16 alkyl, C _7-15 alkyl, _{C 7-14} alkyl _, C _7-13 alkyl, C _7-12 alkyl, C _7-11 alkyl, C 7-10 alkyl, C _7-9 alkyl, C _7-8 alkyl, C _8-20 alkyl, C 8-19 alkyl, C 8-18 alkyl, C _8-17 alkyl, C 8-16 alkyl, C 8-15 alkyl, C _8-14 alkyl, C _8-13 alkyl, C _8-12 alkyl, C _8-11 alkyl, C _8-10 alkyl, C _{8-9 alkyl, C 8-8 alkyl, C 8-20 alkyl ,} C _8-19 alkyl, C 8-18 alkyl, C _8-17 alkyl, C 8-16 alkyl, C _8-15 alkyl, C _8-14 alkyl, C _8-13 alkyl, C _8-12 alkyl, C _8-11 alkyl, C _8-10 alkyl, C _{C 8-9} alkyl, C _9-20 alkyl, C _9-19 alkyl, C _9-18 alkyl, C _9-17 alkyl, C _9-16 alkyl, C _9-15 alkyl, C _9-14 alkyl, C _9-13 alkyl, C _9-12 alkyl, C _9-11 alkyl, C _9-10 alkyl, C _10-20 alkyl, C _10-19 alkyl, C _10-18 alkyl, C _10-17 alkyl, C _10-16 alkyl, C _10-15 alkyl, C _{10-14 alkyl} , C _10-13 alkyl, C _10-12 alkyl, C _10-11 alkyl, C _11-20 alkyl, C _11-19 alkyl, C _11-18 alkyl, C _11-17 alkyl, C _11-16 alkyl, C _11-15 alkyl, C C _11-14 alkyl, C _11-13 alkyl, C _11-12 alkyl, C _12-20 alkyl, C _12-19 alkyl, C _12-18 alkyl, C _12-17 alkyl, C _12-16 alkyl, C _12-15 alkyl, C _{12- 14} alkyl, C _12-13 alkyl, C _13-20 alkyl, C _13-19 alkyl, C _13-18 alkyl, C _13-17 alkyl, C _13-16 alkyl, C _{13-15 alkyl, C 13-14 alkyl} , C _14-20 alkyl, C _14-19 alkyl, C _14-18 alkyl, C _14-17 alkyl, C _14-16 alkyl, C _14-15 alkyl, C _{15-20 alkyl, C 15-19 alkyl} , C _15-18 alkyl, C C _15-17 alkyl, C _15-16 alkyl, C _16-20 alkyl, C _16-19 alkyl, C _16-18 alkyl, C _16-17 alkyl, C _17-20 alkyl, C _17-19 alkyl, C _{17- 18} alkyl, C _16-17 alkyl, C _18-20 alkyl, C _18-19 alkyl, C _19-20 alkyl, C ₂₀ alkyl, C ₁₉ alkyl, C ₁₈ alkyl, C ₁₇ alkyl, C ₁₆ alkyl, C ₁₅ alkyl, C ₁₄ alkyl, C ₁₃ alkyl, C ₁₂ alkyl, C ₁₁ alkyl, C ₁₀ alkyl _, C 8 alkyl, C ₇ alkyl, C ₆ alkyl, C ₅ alkyl, C ₄ alkyl, C ₃ alkyl, ethyl, methyl.

Unless otherwise specified, the term "C _1-3 alkyl" is used to represent a straight or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of C _1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

Unless otherwise specified, the term "C _1-6 alkyl" is used to represent a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. _C1-6 alkyl includes _C1-2 , _C1-3 , _C2-3 and _C1-4 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). Examples of _C1-6 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl, etc.

Unless otherwise specified, " _C2-6 alkenyl" is used to represent a straight or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon double bond, which may be located at any position of the group. The _C2-6 alkenyl group includes _C2-4 , _C2-3 , _C4 , _C3 and _C2 alkenyl, etc.; it may be monovalent, divalent or polyvalent. Examples of _C2-6 alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, etc.

Unless otherwise specified, "C _2-6 alkynyl" is used to represent a straight or branched hydrocarbon group consisting of 2 to 6 carbon atoms containing at least one carbon-carbon triple bond, which may be located at any position of the group. The C _2-6 alkynyl group includes C _2-4 , C _2-3 , C ₄ , C ₃ and C ₂ alkynyl groups, etc. It may be monovalent, divalent or polyvalent. Examples of C _2-6 alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, etc.

Unless otherwise specified, the term "C _1-6 alkoxy" by itself or in combination with other terms, means an alkyl group containing 1 to 6 carbon atoms connected to the rest of the molecule through an oxygen atom. The C _1-6 alkoxy includes C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ , C ₅ , C ₄ and C ₃ alkoxy, etc. Examples of _{C 1-6} alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), pentoxy (including n-pentoxy, isopentyl and neopentyl), hexyl and the like.

Unless otherwise specified, the term "C _1-3 alkoxy" by itself or in combination with other terms refers to those alkyl groups containing 1 to 3 carbon atoms connected to the rest of the molecule through an oxygen atom. The C _1-3 alkoxy includes C _1-2 , C _2-3 , C ₃ and C ₂ alkoxy, etc. Examples of C _1-3 alkoxy include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

Unless otherwise specified, the term "C _1-6 alkylamino" by itself or in combination with other terms refers to those alkyl groups containing 1 to 6 carbon atoms attached to the rest of the molecule through an amino group. The C _1-6 alkylamino group includes C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ , C ₅ , C ₄ , C ₃ and C ₂ alkylamino groups, etc. Examples of C _1-6 alkylamino groups include, but are not limited to, -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -N(CH ₂ CH 3 )(CH ₂ CH ₃ ), -NHCH ₂ CH ₂ CH ₃ , -NHCH ₂ (CH ₃ ) ₂ , -NHCH _{2 CH 2} CH ₂ CH _3, and the like.

Unless otherwise specified, the term "C _1-3 alkylamino" by itself or in combination with other terms, refers to those alkyl groups containing 1 to 3 carbon atoms attached to the rest of the molecule through an amino group. The C _1-3 alkylamino group includes C _1-2 , C ₃ and C ₂ alkylamino groups, etc. Examples of C _1-3 alkylamino groups include, but are not limited to, -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -NHCH ₂ CH ₂ CH ₃ , -NHCH ₂ (CH ₃ ) ₂ , and the like.

Unless otherwise specified, "C _3-20 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 20 carbon atoms, including monocyclic, bicyclic and tricyclic systems, wherein the bicyclic and tricyclic systems include spirocyclic, cyclic and bridged rings. The C _3-12 cycloalkyl includes C _3-10 , C _3-8 , C _3-6 , C _3-5 , C _4-10 , C _4-8 , C _4-6 , C _4-5 , C _5-8 and C _5-6 cycloalkyl, etc.; it can be monovalent, divalent or polyvalent. Examples of C _3-12 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2] bicyclooctane, [4.4.0] bicyclodecane, etc.

Unless otherwise specified, "C _3-6 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, including monocyclic and bicyclic systems, wherein the bicyclic system includes spirocyclic, fused and bridged rings. The C _3-8 cycloalkyl includes C _3-6 , C _3-5 , C _4-8 , C _4-6 , C _4-5 or C _5-6 cycloalkyl, etc.; it can be monovalent, divalent or polyvalent. Examples of C _3-8 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2] bicyclooctane, etc.

Unless otherwise specified, the term "3-6 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated cyclic group consisting of 3 to 6 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., C(=O), NO and S(O) _p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spiro, fused and bridged rings. In addition, with respect to the "4-7 membered heterocycloalkyl", heteroatoms may occupy the position at which the heterocycloalkyl is connected to the rest of the molecule. The 3-6 membered heterocycloalkyl includes 3-4 membered, 3-5 membered, 4-5 membered, 4-6 membered, 5-6 membered, 3 membered, 4 membered, 5 membered, 6 membered heterocycloalkyl, etc. Examples of 3-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl, alkyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, etc.

Unless otherwise specified, the term "4-6 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated cyclic group consisting of 4 to 6 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the carbon atoms are optionally oxoed (i.e., C(O)), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) _p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring systems include spirocyclic, paracyclic and bridged rings. In addition, with respect to the "4-6 membered heterocycloalkyl", heteroatoms may occupy the position where the heterocycloalkyl is connected to the rest of the molecule. The 4-6 membered heterocycloalkyl includes 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyls, etc. Examples of 4-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl or hexahydropyridazinyl, etc.

Unless otherwise specified, the term "4-7 membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 4 to 7 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., C(=O), NO and S(O) _p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spirocyclic, paracyclic and bridged rings. In addition, with respect to the "4-7 membered heterocycloalkyl", heteroatoms may occupy the position at which the heterocycloalkyl is connected to the rest of the molecule. The 4-7 membered heterocycloalkyl includes 4-6 membered, 4-5 membered, 5-6 membered, 6-7 membered, 4 membered, 5 membered, 6 and 7 membered heterocycloalkyl, etc. Examples of 4-7 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl or dioxepanyl, etc.

In some embodiments of the present invention, 3-20 membered heterocycloalkyl includes but is not limited to 20 membered, 18 membered, 16 membered, 10 membered, 8-16 membered, 8-12 membered, 6-10 membered, 5-8 membered, 14-6 membered, 4-7 membered, 5-6 membered, 5-7 membered, 5-8 membered, 6-7 membered, 6-8 membered, 7-8 membered, 4 membered, 5 membered, 6 membered, 7 membered, and 8 membered heterocycloalkyl. In some embodiments of the present invention, the 3-20 membered heterocycloalkyl group includes, but is not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl or dioxepanyl, etc.

In some embodiments of the present invention, 8-16 membered heterocycloalkyl includes but is not limited to 16 membered, 10 membered, 8-16 membered, 8-15 membered, 8-14 membered, 8-13 membered, 8-12 membered, 8-11 membered, 8-10 membered, 8-9 membered, 16 membered, 15 membered, 14 membered, 13 membered, 12 membered, 11 membered, 10 membered, 9 membered, and 8 membered heterocycloalkyl. In some embodiments of the present invention, examples of 4-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, etc.

Unless otherwise specified, the term "8-16 membered heterocycloalkenyl" by itself or in combination with other terms means a partially unsaturated cyclic group consisting of 8 to 16 ring atoms containing at least one carbon-carbon double bond, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms may be optionally oxidized (i.e., C(=O), NO and S(O)p, p is 1 or 2). It includes monocyclic, bicyclic and tricyclic ring systems, wherein the bicyclic and tricyclic ring systems include spirocyclic, fused and bridged rings, and any ring of this system is non-aromatic. In addition, with respect to the "8-16 membered heterocycloalkenyl", heteroatoms may occupy the linking portion of the heterocycloalkenyl group to the rest of the molecule. The 8-16 membered heterocycloalkenyl group includes 8-10 membered, 8-12 membered, 8-14 membered, 8 membered, 9 membered, 10 membered, 12 membered and 16 membered heterocycloalkenyl groups. Examples of 8-16 membered heterocycloalkenyl groups include, but are not limited to

Unless otherwise specified, the terms "8-16 membered heteroaromatic ring" and "8-16 membered heteroaryl" are used interchangeably in the present invention. The term "8-16 membered heteroaryl" means a cyclic group consisting of 8 to 16 ring atoms and containing a conjugated π electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms are optionally oxidized (i.e., C(=O), NO and S(O)p, p is 1 or 2). It can be a monocyclic, fused bicyclic or fused tricyclic system, wherein at least one ring is aromatic. The 8-16 membered heteroaryl can be connected to the rest of the molecule through a heteroatom or a carbon atom. The 8-16 membered heteroaryl includes 8-10 membered, 8-12 membered, 10 membered, 8 membered heteroaryl, etc. Examples of the 8-16 membered heteroaryl include, but are not limited to, benzimidazolyl (including 2-benzimidazolyl, etc.), benzoxazolyl, indolyl (including 5-indolyl, etc.), isoquinolyl (including 1-isoquinolyl and 5-isoquinolyl, etc.), quinoxalinyl (including 2-quinoxalinyl and 5-quinoxalinyl, etc.), quinolyl (including 3-quinolyl and 6-quinolyl, etc.),

Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" of the present invention can be used interchangeably. The term "5-6 membered heteroaryl" means a monocyclic group consisting of 5 to 6 ring atoms with a conjugated π electron system, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen and sulfur heteroatoms are optionally oxidized (i.e., C(=O), NO and S(O)p, p is 1 or 2). The 5-6 membered heteroaryl can be connected to the rest of the molecule via a heteroatom or a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl) and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

The compounds invented herein can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments invented herein.

The structure of the compounds invented herein can be confirmed by conventional methods known to those skilled in the art. If the invention herein involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data of the cultivated single crystal using a Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is: /ω scanning, after collecting relevant data, the direct method (Shelxs97) is further used to analyze the crystal structure to confirm the absolute configuration.

The solvents used in the invention herein are commercially available.

The present invention uses the following abbreviations: Boc represents tert-butyloxycarbonyl, which is an amine protecting group; DMF represents N,N-dimethylformamide; NBS represents N-bromosuccinimide; HATU represents O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; HPLC represents high pressure liquid chromatography; LCMS represents liquid chromatography-mass spectrometry; DMSO-d ₆ represents deuterated dimethyl sulfoxide; CD ₃ OD represents deuterated methanol; CDCl ₃ represents deuterated chloroform; CD ₃ CN represents deuterated acetonitrile.

DETAILED DESCRIPTION

The present invention is described in detail below by way of examples, but it is not intended to impose any adverse limitations on the present invention. The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention. For those skilled in the art, it is not necessary to describe the specific embodiments of the present invention without departing from the spirit and scope of the present invention. It will be apparent that various changes and modifications can be made to the formula.

Intermediate A

Synthesis route:

Step 1:

Compound A-2 (941.79 μL, 4.20 mmol), triethylamine (1.59 mL, 11.45 mmol), dichlorobis(triphenylphosphine)palladium (267.86 mg, 381.62 μmol) and cuprous iodide (145.36 mg, 763.23 μmol) were added to a solution of compound A-1 (1 g, 3.82 mmol) in tetrahydrofuran (20 mL). The reaction solution was stirred at 30°C for 12 h under nitrogen protection. After the reaction was completed, 50 mL of water was added to the reaction solution, and after stirring for 5 min, the aqueous phase was extracted with ethyl acetate (50 mL×2), washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by column chromatography (silica gel column, eluent gradient 0-15% petroleum ether/ethyl acetate) to obtain compound A-3. MS (ESI) m/z: 317.2 [M+H] ⁺ .

Step 2:

At 0°C, lithium deuterated tetrahydrofuran (59.96 mg, 1.58 mmol, 81.46 μL) was added to a solution of compound A-3 (0.5 g, 1.58 mmol) in tetrahydrofuran (10 mL), and the mixture was stirred at 0°C for 0.5 h. After the reaction was completed, the reaction solution was carefully quenched with saturated aqueous ammonium chloride solution (20 mL), extracted with ethyl acetate (50 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain intermediate A. MS (ESI) m/z: 273.2 [M+H ⁺ -18].

Intermediate B

Synthesis route:

Step 1:

Compound triphenylmethane (19.45 g, 69.76 mmol) was added to a dichloromethane (100 mL) solution of compound B-1 (10 g, 46.51 mmol), and triethylamine (12.95 mL, 93.01 mmol) was slowly added dropwise to the mixture, and the temperature was kept at 25 ° C and stirred for 2 h. After the reaction was completed, 200 mL of water was added to the reaction solution, stirred for 5 min, and allowed to stand for separation. The aqueous phase was extracted with dichloromethane (2 times, 200 mL each time), and the organic phases were combined, washed with saturated brine twice, 200 mL each time, dried over anhydrous sodium sulfate, filtered and concentrated. Ethyl acetate (45 mL) was added to the obtained residue, stirred at 25 ° C for 30 min, the filter cake was collected, and dried to obtain compound B-2.

Step 2:

Under nitrogen protection, compound B-2 (10 g, 21.54 mmol) was added to tetrahydrofuran (100 mL), cooled to -78 °C, N, N-diisopropylamide lithium (2 mol/L, 16.16 mL) was slowly added dropwise, and the reaction was stirred at -78 °C for 0.5 h. Then iodomethane (4.59 g, 32.31 mmol, 2.01 mL) was slowly added dropwise to the reaction solution. After the addition was completed, the reaction solution was heated to 15 °C and stirred at 15 °C for 1.5 h. The reaction was completed. Then, 200 mL of water was added to the reaction solution, and after stirring for 5 min, it was extracted with ethyl acetate (2 times, 200 mL each time), the organic phases were combined, washed with saturated brine (2 times, 100 mL each time), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by high performance liquid chromatography (chromatographic column: Kromasil Eternity XT 250*80mm*10μm; mobile phase: [water (ammonia water)-acetonitrile]; gradient: acetonitrile 65%-95%) to obtain compound B-3.

¹ H NMR (400MHz, CDCl ₃ ) δ = 7.85 (s, 1H), 7.52-7.30 (m, 10H), 7.25-7.15 (m, 6H), 2.42 (d, J = 2.4Hz, 3H).

Step 3:

To a solution of compound B-3 (5 g, 10.61 mmol) in 1,4-dioxane (250 mL) were added bis-naphthalene borate (5.39 g, 21.22 mmol), potassium acetate (3.12 g, 31.82 mmol), [1,1-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane (866.25 mg, 1.06 mmol), and the mixture was stirred at 100 ° C for 16 h under a nitrogen atmosphere. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 5/1) to obtain intermediate B.

Intermediate C

Synthesis route:

Step 1:

Compound C-1 (5 g, 12.99 mmol) was added to a solution of N,N-diisopropylethylamine (8.39 g, 64.95 mmol) in phosphorus oxychloride (25 mL) at 0°C, and the reaction solution was stirred at 110°C for 4 h. After the reaction was completed, the mixture was concentrated under reduced pressure to obtain compound C-2. MS (ESI) m/z: 422.8 [M+H] ⁺ .

Step 2:

Compound C-3 (3.09 g, 15.59 mmol) and N,N-diisopropylethylamine (5.04 g, 38.97 mmol) were added to a solution of compound C-2 (5.48 g, 12.99 mmol) in 1,4-dioxane (55 mL), and the reaction solution was stirred at 25°C for 16 h. After the reaction was completed, 200 mL of water was added to the reaction solution, stirred for 5 min, extracted with dichloromethane (2 times, 200 mL each time), the organic phases were combined, washed with saturated brine (2 times, 200 mL each time), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (silica gel column, eluent gradient 0-50% petroleum ether/ethyl acetate) to obtain intermediate C. MS (ESI) m/z: 584.9 [M+H] ⁺ .

Intermediate D

Synthesis route:

Step 1:

Compound D-2 (8.43 g, 37.95 mmol), potassium carbonate (8.74 g, 63.25 mmol) and 1,1-di(tert-butylphosphino)ferrocenepalladium chloride (2.06 g, 3.16 mmol) were added to a mixed solvent of compound D-1 (10 g, 31.63 mmol), and the mixed system was stirred at 100 ° C for 12 h under a nitrogen atmosphere. After the reaction was completed, the reaction solution was concentrated to dryness, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate = 1/1 to 0/1) to obtain compound D-3. MS (ESI) m/z: 332.2 [M+H] ⁺ .

Step 2:

To a solution of compound D-3 (10 g, 28.89 mmol) in ethyl acetate (10 mL) was added a hydrogen chloride/ethyl acetate solution (4 mol/L, 20 mL), and the mixture was stirred at 25°C for 1 h. After the reaction was completed, the mixture was filtered, the filter cake was collected, and dried to obtain the hydrochloride of compound D-4. MS (ESI) m/z: 232.2 [M+H] ⁺ .

Step 3:

Compound D-5 (7.02 g, 30.34 mmol), O-(7-azabenzotriazole-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (13.18 g, 34.67 mmol), and triethylamine (14.94 g, 115.58 mmol) were added to a solution of the hydrochloride of compound D-4 (2.6 g) in N,N-dimethylformamide (100 mL), and the mixture was stirred at 25°C for 2 h. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by high performance liquid chromatography (chromatographic column: Welch Ultimate XT-CN 250*50mm*10μm; mobile phase: [n-hexane-ethanol (0.1% trifluoroacetic acid)]; gradient: acetonitrile 5%-45%) to obtain compound D-6. MS (ESI) m/z: 445.3 [M+H] ⁺ .

Step 4:

To a solution of compound D-6 (2 g, 4.5 mmol) in ethyl acetate (5 mL) was added hydrogen chloride/ethyl acetate solution (4 mol/L, 5 mL), and the mixture was stirred at 25°C for 1 h. After the reaction was completed, the mixture was filtered, the filter cake was collected, and dried to obtain the hydrochloride of compound D-7. MS (ESI) m/z: 345.2 [M+H] ⁺ .

Step 5:

Compound D-8 (8.39 g, 38.60 mmol), O-(7-azabenzotriazole-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (16.77 g, 44.11 mmol) and triethylamine (16.63 g, 128.65 mmol) were added to a solution of the hydrochloride of compound D-7 (14 g) in N,N-dimethylformamide (100 mL), and the mixture was stirred at 25°C for 2 h. The reaction solution was concentrated, and the crude product was purified by preparative high performance liquid chromatography (chromatographic column: Phenomenex luna C18 250*70mm*10μm; mobile phase: [water (formic acid)-acetonitrile]; gradient: acetonitrile 36%-56%) to obtain compound D-9. MS (ESI) m/z: 544.4 [M+H] ⁺ .

Step 6:

To a solution of compound D-9 (2 g, 3.68 mmol) in ethyl acetate (5 mL) was added hydrogen chloride/ethyl acetate solution (4 mol/L, 5.2 mL), and the mixture was stirred at 25°C for 1 h. After the reaction was completed, the mixture was filtered, the filter cake was collected, and dried to obtain the hydrochloride of compound D-10. MS (ESI) m/z: 444.2 [M+H] ⁺ .

Step 7:

Potassium carbonate (1.49 g, 10.78 mmol) and copper sulfate pentahydrate (76.90 mg, 308 μmol) were added to a solution of the hydrochloride of compound D-10 (1.6 g) in methanol (35 mL) at 0°C, and then the hydrochloride of compound D-11 (1.61 g, 7.7 mmol) was slowly added to the system at 0°C, and the mixed system was stirred at 25°C for 16 h. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (dichloromethane/methanol=20/1 to 10/1) to obtain intermediate D. MS (ESI) m/z: 473.2 [M+H] ⁺ .

Example 1

Synthesis route:

Step 1:

To a mixed solution of intermediate C (5 g, 8.57 mmol) in tetrahydrofuran (20 mL) and N,N-dimethylformamide (20 mL) were added triethylenediamine (192.20 mg, 1.71 mmol), cesium carbonate (8.37 g, 25.70 mmol) and compound 1-1 (4.37 g, 42.84 mmol), and the mixed system was stirred at 25°C for 24 h. After the reaction was completed, 200 mL of water was added to the reaction solution, stirred for 5 min, extracted with ethyl acetate (2 times, 200 mL each time), the organic phases were combined, washed with saturated brine (3 times, 100 mL each time), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column chromatography (silica gel column, eluent gradient 0-50% petroleum ether/ethyl acetate, 100 mL/min) to obtain compound 1-2. MS (ESI) m/z: 649.1 [M+H] ⁺ .

Step 2:

To a mixed solution of compound 1-2 (2.8 g, 4.31 mmol) in toluene (30 mL) and water (6 mL) was added [1,1-bis(diphenylphosphine) Ferrocene] palladium dichloride dichloromethane (352.17 mg, 431.24 μmol), compound 1-3 (740.85 mg, 8.62 mmol) and potassium carbonate (1.19 g, 8.62 mmol), the reaction solution was stirred at 80°C for 12 h under nitrogen protection. After the reaction was completed, water (100 mL) was added to the reaction solution, and after stirring for 5 min, the aqueous phase was extracted with ethyl acetate (2 times, 100 mL each time), the organic phases were combined, washed with saturated brine (2 times, 100 mL each time), the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent gradient 0-50% petroleum ether/ethyl acetate, 100 mL/min) to obtain compound 1-4. MS (ESI) m/z: 565.2 [M+H] ⁺ .

Step 3:

Sodium hydrogen (119.26 mg, 2.98 mmol, 60% purity) was added to a solution of intermediate A (866.18 mg, 2.98 mmol) in 1,4-dioxane (40 mL), and the mixture was stirred at 25°C for 2 h. Compound 1-4 (1.4 g, 2.48 mmol) was then added to the mixture, and the reaction solution was stirred at 25°C for 12 h under nitrogen protection. After the reaction was completed, water (100 mL) was added to the reaction solution, and after stirring for 5 min, the aqueous phase was extracted with ethyl acetate (2 times, 100 mL each time), the organic phases were combined, washed with saturated brine (2 times, 100 mL each time), the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated, and the crude product was purified by silica gel column chromatography (eluent gradient 0-30% petroleum ether/ethyl acetate, 100 mL/min) to obtain compound 1-5. MS (ESI) m/z: 835.4 [M+H] ⁺ .

Step 4:

Potassium phosphate (419.98 mg, 1.98 mmol) and tetrakis(triphenylphosphine)palladium (76.21 mg, 65.95 μmol) were added to a mixed solution of compound 1-5 (0.55 g, 659.50 μmol) and intermediate B (410.28 mg, 791.40 μmol) in 1,4-dioxane (4 mL) and water (1 mL). The mixed system was stirred at 110°C for 16 h under nitrogen protection. After the reaction was completed, water (100 mL) was added to the reaction solution, and after stirring for 5 min, the aqueous phase was extracted with ethyl acetate (2 times, 100 mL each time), the organic phases were combined, washed with saturated brine (2 times, 50 mL each time), and the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 1-6. MS (ESI) m/z: 1145.7 [M+H] ⁺ .

Step 5:

Tetrabutylammonium fluoride (1 mol/L tetrahydrofuran solution, 10.95 mL) was added to a solution of compound 1-6 (0.75 g, 654.73 μmol) in tetrahydrofuran (10 mL), and the mixture was stirred at 25°C for 10 min. After the reaction was completed, the crude product was concentrated under reduced pressure and purified by silica gel column chromatography (eluent gradient 0-50% petroleum ether/ethyl acetate, 100 mL/min) to obtain compound 1-7. MS (ESI) m/z: 989.4 [M+H] ⁺ .

Step 6:

Hydrogen chloride/ethyl acetate (4 mol/L, 10 mL) was added to a solution of compound 1-7 (470 mg, 475.14 μmol) in ethyl acetate (10 mL), and the mixture was stirred at 25° C. for 0.5 h. The reaction solution was concentrated, and the crude product was purified by preparative HPLC (chromatographic column Phenomenex Luna C18 150*25 mm*10 μm; mobile phase: [water (formic acid)-acetonitrile]; gradient: 14%-44% acetonitrile) to give the formate salt of compound 1-8A (LCMS retention time: 0.715 min), MS (ESI) m/z: 647.4 [M+H] ⁺ and the formate salt of compound 1-8B (LCMS retention time: 0.738 min), MS (ESI) m/z: 647.4 [M+H] ⁺ .

Step 7:

To a mixed solution of formate (70 mg) of compound 1-8B in tert-butyl alcohol (5 mL) and water (5 mL) were added intermediate D (56.26 mg, 119.06 μmol), sodium ascorbate (21.44 mg, 108.23 μmol) and copper sulfate pentahydrate (27.02 mg, 108.23 μmol). The mixed system was stirred at 50°C for 16 h. The reaction solution was directly concentrated, and the crude product was purified by preparative HPLC (chromatographic column: Waters xbridge 150*25mm*10μm; mobile phase: [water (ammonium bicarbonate)-acetonitrile]: gradient: 35%-65% acetonitrile) to obtain compound 1B. MS (ESI) m/z: 1119.6 [M+H] ⁺ . ¹ HNMR (400MHz, DMSO-d ₆ ) δ = 13.29-12.99 (m, 1H), 9.00 (s, 1H), 8.66 (s, 1H), 8.49 (br d, J = 8.0Hz, 1H), 7.76-7.63 (m, 2H), 7.57-7.35 (m, 7H), 6.88-6.71 (m ,2H),5.34(br d,J＝10.0Hz,1H),5.26-5.15(m,2H),5.09(br s,1H),4.95-4.82(m,2H),4.46(br t,J＝8.4Hz,1H),4.36-4.24(m,2H),3.94-3.84(m,3H),3.84-3.75(m,2H),3.74-3.67(m,1H),3.66-3.56(m,2H),3.42-3.37(m,3H),3.23-3.16(m,1 H),3.09(br d,J＝8.4Hz,1H),2.53-2.53(m,2H),2.48-2.46(m,3H),2.17-1.93(m,7H),1.89 -1.75(m,2H),1.74-1.61(m,2H),1.44-1.29(m,1H),1.17-0.99(m,3H),0.83-0.48(m,7H).

Example 2

Synthesis route:

Step 1:

Intermediate C (5 g, 8.57 mmol) and compound 2-1 (1.33 g, 10.28 mmol) were added to a mixed solution of N, N-dimethylformamide (25 mL) and tetrahydrofuran (25 mL), triethylenediamine (192.2 mg, 1.71 mmol) and cesium carbonate (8.37 g, 25.7 mmol) were added to the system, and the mixed system was stirred at 25 ° C for 12 h. The reaction solution was diluted with water (100 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic layer was washed with saturated brine (50 mL × 3), dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to obtain a crude compound 2-2, which was directly used in the next step. MS (ESI) m/z: 696.0 [M + H] ⁺ .

Step 2:

Compound 2-2 (5 g, 7.39 mmol), cyclopropylboronic acid (952.56 mg, 11.09 mmol), potassium carbonate (3.07 g, 22.18 mmol) and [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane (603.74 mg, 739.30 μmol) were added to a mixed solution of anhydrous toluene (40 mL) and water (8 mL), replaced with nitrogen three times, and the mixed system was stirred at 80 ° C for 12 h. Water (100 mL) was added to the reaction solution for dilution, extracted with ethyl acetate (50 mL × 2), the organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to obtain compound 2-3. MS (ESI) m/z: 590.2 [M + H] ⁺ .

Step 3:

Intermediate A (2g, 6.88mmol) was dissolved in anhydrous dioxane (30mL) and replaced with nitrogen three times. The system was cooled to 0°C, sodium hydrogen (609.61mg, 15.24mmol, 60% purity) was added in batches and stirred for 1h, and then compound 2-3 was added to the reaction system, and the mixed system was stirred at 25°C for 12h. The reaction solution was quenched with saturated ammonium chloride (50mL), diluted with water (150mL), extracted with ethyl acetate (100mL×2), combined with organic phases, washed with saturated brine (100mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silicon dioxide, petroleum ether/ethyl acetate = 10/1 to 1/10) to obtain compound 2-4. MS (ESI) m/z: 862.1 [M+H] ⁺ .

Step 4:

Compound 2-4 (1.8 g, 2.06 mmol), intermediate B (1.19 g, 2.3 mmol), dicyclohexylphosphine-2,6-dimethoxybiphenyl (171.65 mg, 418.12 μmol), potassium phosphate (1.33 g, 6.27 mmol) and tris(dibenzylideneacetone)dipalladium (191.44 mg, 209.06 μmol) were dissolved in a mixed solution of dioxane (30 mL) and water (6 mL), the reaction solution was replaced with nitrogen three times, and the mixed system was stirred at 110 ° C for 12 h. Water (100 mL) was added to the reaction solution for dilution, extracted with ethyl acetate (50 mL × 2), the organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and dried to obtain a crude product, which was purified by column chromatography (silica, petroleum ether/ethyl acetate = 10/1 to 1/10) to obtain compound 2-5. MS (ESI) m/z: 1171.8 [M+H] ⁺ .

Step 5:

Tetrabutylammonium fluoride (1 mol/L tetrahydrofuran solution, 16 mL) was added to a solution of compound 2-5 (1.6 g, 1.36 mmol) in tetrahydrofuran (16 mL), and the mixture was stirred at 25°C for 0.5 h. Water (100 mL) was added to the reaction solution for dilution, and the mixture was extracted with ethyl acetate (50 mL×2). The organic phases were combined, washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was purified by column chromatography (silica, dichloromethane/methanol=50/1) to obtain compound 2-6. MS (ESI) m/z: 1015.9 [M+H] ⁺ .

Step 6:

Hydrogen chloride/ethyl acetate (4 mol/L, 10 mL) was added to a solution of compound 2-6 (1.3 g, 1.28 mmol) in ethyl acetate (10 mL), and the mixture was stirred at 25° C. for 0.5 h. The reaction solution was concentrated to obtain a crude product, which was subjected to preparative high performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150*40mm*15μm; mobile phase: [water (hydrochloric acid)-acetonitrile]; gradient: acetonitrile 15%-45%) to obtain the hydrochloride of compound 2-7A (LCMS retention time: 0.707 min), MS (ESI) m/z: 674.2 [M+H] ⁺ and the hydrochloride of compound 2-7B (LCMS retention time: 0.727 min), MS (ESI) m/z: 674.2 [M+H] ⁺ .

Step 7:

The hydrochloride of compound 2-7B (150 mg), intermediate D (105.2 mg, 222.63 μmol), copper sulfate pentahydrate (55.59 mg, 222.63 μmol) and sodium ascorbate (44.10 mg, 222.63 μmol) were added to a mixed solution of tert-butyl alcohol (7.5 mL) and water (7.5 mL), replaced with nitrogen three times, and the mixed system was stirred at 50° C. for 12 h. The reaction solution was concentrated, and the crude product was subjected to preparative high performance liquid chromatography (column: Waters Xbridge C18 150*40mm*15μm; mobile phase: [water (hydrochloric acid)-acetonitrile]; gradient: acetonitrile 15%-45%) to obtain the hydrochloride of compound 2B. MS (ESI) m/z: 1145.7 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δppm 10.36 (br dd, J＝6.4, 2.81Hz, 1H), 9.85-9.46 (m, 1H), 9.16 (s, 1H), 8.72-8.56 (m, 2H), 7.65 (br d, J＝8.0Hz, 2H), 7.54-7.36 (m, 7H),6.63(br d,J＝6.4Hz,2H),5.90-5.77(m,1H),5.58(br d,J＝12.0Hz,1H),5.33(br d,J＝10.0Hz,1H),4.88-4.79(m,1H),4.65(br s,3H),4.49-4.43(m,2 H),3.93(br d,J＝4.0Hz,1H),3.90-3.75(m,4H),3.75-3.65(m,3H),3.66-3.56(m,4H),3.52(br d,J＝16.0Hz,1H),3.446-3.36(m,1H),2.48(s,3H),2.23-2.39(m, 3H),2.18-2.06(m,2H),2.05-1.94(m,6H),1.86-1.73(m,1H),1.49-1.38(m,1H),1.30-1.00(m,4H),0.72(br d,J＝6.4Hz,6H).

Example 3

Synthesis route:

Step 1:

Concentrated sulfuric acid (64.11 mg, 653.70 μmol, 34.84 μL) was added to a solution of compound 3-1 (10 g, 65.37 mmol) in ethanol (30 mL), and the reaction solution was stirred at 80° C. for 16 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, 20 mL of ethyl acetate was added to the reaction solution for dilution, the organic phase was washed with saturated sodium bicarbonate (20 mL×2), washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate, and the filtrate was concentrated and dried to obtain compound 3-2.

Step 2:

Potassium carbonate (20.16 g, 145.83 mmol) was added to a solution of compound 3-2 (4.4 g, 24.31 mmol) and compound 3-3 (11.85 g, 121.53 mmol) in acetonitrile (40 mL), and the mixture was stirred at 80° C. for 16 h. The reaction solution was filtered, and the filtrate was concentrated to obtain compound 3-4.

Step 3:

At 0°C, lithium aluminum tetrahydride tetrahydrofuran solution (2.5 mol/L, 5.96 mL) was added to a solution of compound 3-4 (2.4 g, 14.89 mmol) in tetrahydrofuran (10 mL), and the mixture was stirred at 0°C for 1 h. The reaction solution was carefully quenched with saturated ammonium chloride solution (40 mL), extracted with ethyl acetate (50 mL×3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and dried to obtain compound 3-5.

Step 4:

To a mixed solution of intermediate C (5 g, 8.57 mmol) in tetrahydrofuran (20 mL) and N,N-dimethylformamide (20 mL) were added triethylenediamine (192.20 mg, 1.71 mmol), cesium carbonate (8.37 g, 25.70 mmol) and compound 3-5 (1.12 g, 9.42 mmol), and the reaction solution was stirred at 25°C for 12 h. After the reaction was completed, 100 mL of water was added to the reaction solution, stirred for 5 min, extracted with ethyl acetate (200 mL×2), the organic phases were combined, washed with saturated brine (100 mL×3), dried over anhydrous sodium sulfate, filtered, and concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=2/1 to 3/2) to obtain compound 3-6. MS (ESI) m/z: 666.1 [M+H] ⁺ .

Step 5:

Cyclopropylboric acid (580.11 mg, 6.75 mmol), [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride (367.68 mg, 450.23 μmol) and potassium carbonate (1.24 g, 9.00 mmol) were added to a mixed solution of compound 3-6 (3 g, 4.50 mmol) in toluene (30 mL) and water (6 mL). The mixed system was stirred at 80 ° C for 12 h under a nitrogen atmosphere. After the reaction was completed, 50 mL of water was added to the reaction solution, stirred for 5 min, extracted with ethyl acetate (50 mL × 2), the organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column (petroleum ether/ethyl acetate = 2/1 to 1/1) to obtain compound 3-7. MS (ESI) m/z: 580.2 [M + H] ⁺ .

Step 6:

Compound 3-7 (1.20 g, 4.13 mmol) and sodium hydrogen sulfide (413.41 mg, 10.34 mmol, 60% purity) were added to a solution of intermediate A (2 g, 3.45 mmol) in tetrahydrofuran (40 mL), and the mixture was stirred at 40°C for 12 h. After the reaction was completed, water (100 mL) was added to the reaction solution, and after stirring for 5 min, the aqueous phase was extracted with ethyl acetate (100 mL×2), the organic phases were combined, washed with saturated brine (100 mL×2), and the organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether/ethyl acetate=2/1 to 3/2) to obtain compound 3-8. MS (ESI) m/z: 852.3 [M+H] ⁺ .

Step 7:

To a mixed solution of compound 3-8 (1.5 g, 1.76 mmol) in 1,4-dioxane (4 mL) and water (1 mL) were added intermediate B (1.01 g, 1.94 mmol), potassium phosphate (1.12 g, 5.29 mmol) and tetrakis(triphenylphosphine)palladium (203.68 mg, 176.26 μmol). The mixed system was stirred at 110°C for 16 h under nitrogen protection. After the reaction was completed, water (50 mL) was added to the reaction solution, and after stirring for 5 min, the aqueous phase was extracted with ethyl acetate (50 mL×2), the organic phases were combined, washed with saturated brine (50 mL×2), and the organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 3-9. MS (ESI) m/z: 1162.6 [M+H] ⁺ .

Step 8:

Tetrabutylammonium fluoride (2 mol/L tetrahydrofuran solution, 2 mL) was added to a solution of compound 3-9 (2 g, 1.72 mmol) in tetrahydrofuran (10 mL), and the mixture was stirred at 25°C for 10 min. After the reaction was completed, water (50 mL) was added to the reaction solution, and after stirring for 5 min, the aqueous phase was extracted with ethyl acetate (50 mL×2), the organic phases were combined, washed with saturated brine (50 mL×2), and the organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was purified by silica gel column chromatography (eluent: petroleum ether/ethyl acetate=2/1 to 1/1) to obtain compound 3-10. MS (ESI) m/z: 1006.6 [M+H] ⁺ .

Step 9:

Hydrogen chloride/ethyl acetate (4 mol/L, 10 mL) was added to a solution of compound 3-10 (1 g, 993.83 μmol) in ethyl acetate (10 mL), and the mixture was stirred at 25°C for 10 min. The reaction mixture was filtered, the filter cake was collected, and the crude product was purified by preparative high performance liquid chromatography (chromatographic column: Phenomenex Luna C18 150*40mm*15μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 20%-50% acetonitrile) to obtain the trifluoroacetate of compound 3-11A (LCMS retention time: 0.786 min). MS (ESI) m/z: 664.4 [M+H] ⁺ . And the trifluoroacetate of compound 3-11B (LCMS retention time: 0.806 min). MS (ESI) m/z: 664.4 [M+H] ⁺ .

Step 10:

To a mixed solution of the trifluoroacetate salt of compound 3-11B (0.36 g) in tert-butanol (36 mL) and water (36 mL) were added intermediate D (281.92 mg, 596.58 μmol), sodium ascorbate (107.44 mg, 542.35 μmol) and copper sulfate pentahydrate (135.42 mg, 542.35 μmol). The mixed system was stirred at 50°C for 2 h. The reaction solution was directly concentrated, and the crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Luna C18 150*40mm*15μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 15%-45% acetonitrile) to obtain the trifluoroacetate salt of compound 3B. MS (ESI) m/z: 1136.6 [M+H] ⁺ .

¹ HNMR (400MHz, DMSO-d ₆ )δ=13.71-12.57(m,1H),9.40-9.14(m,1H),9.00(s,1H),8.66(s,1H),8.55-8.33(m, 2H),7.74-7.59(m,2H),7.49-7.34(m,7H),6.75(d,J＝7.2Hz,2H),5.34(d,J＝10.4Hz,1H),5.24(br s, 1H),4.87-4.82(m,1H),4.66-4.54(m,4H),4.49-4.42(m,4H),4.35-4.29(m,4H),4.10(br d,J＝10.4Hz,2H),3.79(br dd,J＝4.4,10.8Hz,1H),3.70-3.60(m,3H),3.48-3.42(m,1H),3.18-3.09(m,1H ),2.57(s,3H),2.47(s,3H),2.29(br d,J＝10.0Hz,1H),2.12-2.06(m,1H),2.05-1.97(m,4H),1.80(ddd ,J＝4.4,8.4,12.8Hz,1H),1.47-1.35(m,1H),1.13-1.06(m,6H),0.79-0.67(m,5H),0.66-0.57(m,2H).

Example 4

Synthesis route:

Step 1:

At 20°C, potassium carbonate (283.37 g, 2.05 mol) was added to a solution of compound 3-3 (100 g, 1.03 mol) in N,N-dimethylformamide (500 mL), and the mixture was distilled at 100-130°C to collect product 4-1. When the product in the receiving bottle no longer increased, the reaction was completed. ¹ H NMR (400 MHz, CD ₃ Cl) δppm 3.52 (s, 3H) 2.71 (s, 3H) 2.41 (s, 1H).

Step 2:

Compound 4-2 (15.02 g, 245.97 mmol) was added to an isopropanol solution of compound 4-1 (15 g, 258.27 mol) at 20°C, and the mixture was stirred at 35°C for 16 h. After the reaction, the volatile impurities were removed by concentration under reduced pressure at 30°C, and the product 4-3 was distilled out using an oil pump at 25°C. ¹ H NMR (400 MHz, CD ₃ CN) δppm 3.88-3.85 (m, 1H) 3.47 (s, 3H) 2.57-2.48 (m, 5H) 1.08 (d, 3H).

Step 3:

At 20°C, triethylenediamine (307.52 mg, 2.74 mmol) and cesium carbonate (13.40 g, 41.12 mmol) were added to a solution of compound 4-3 (8 g, 13.71 mmol) and intermediate C (6.53 g, 54.83 mmol) in tetrahydrofuran (40 mL) and N,N-dimethylformamide (40 mL), and the mixture was stirred at 20°C for 16 h under nitrogen protection. The reaction solution was poured into water (200 mL), and a crude product was precipitated, filtered, and the filter cake was collected. The crude product was purified by column chromatography (silica gel, eluent, petroleum ether: ethyl acetate = 3:2) to obtain compound 4-4. LCMS (ESI) m/z: 666.0 [M+H] ⁺ .

Step 4:

At 20°C, 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (219.63 mg, 300.16 μmol) and potassium carbonate (829.66 mg, 6.00 mmol) were added to a solution of compound 4-4 (2 g, 3.00 mmol) and cyclopropylboronic acid (567.22 mg, 6.60 mmol) in toluene (40 mL) and water (8 mL), and the mixture was stirred at 80°C for 16 h under nitrogen protection. The reaction solution was poured into water (200 mL), then extracted with ethyl acetate (80 mL*2), and the combined organic phase was washed with water (100 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, petroleum ether: ethyl acetate = 3:2) to obtain compound 4-5. LCMS (ESI) m/z: 580.0 [M+H] ⁺ .

Step 5:

At 20°C, sodium hydrogen (496.09 mg, 12.40 mmol, 60% purity) was added to a tetrahydrofuran (30 mL) solution of compound 4-5 (2.4 g, 4.13 mmol) and intermediate A (1.44 g, 4.96 mmol), and the mixture was stirred at 40°C for 16 h. Water (60 mL) was slowly added to the reaction solution to quench the reaction and stirred for 5 min, then extracted with ethyl acetate (100 mL*3), the combined organic phase was washed with saturated brine (150 mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, petroleum ether: ethyl acetate = 3:1), and then prepared by HPLC (chromatographic column: Phenomenex luna C18 250*80mm*10μm; mobile phase: [water (0.225% formic acid)-(acetonitrile /tetrahydrofuran = 2/1)]; gradient (acetonitrile/tetrahydrofuran = 2/1): 60%-100%) to give compound 4-6. LCMS (ESI) m/z: 850.2 [M+H] ⁺ .

Step 6:

At 20°C, tetrakistriphenylphosphine palladium (516.00 mg, 446.54 μmol) and potassium phosphate (1.42 g, 6.70 mmol) were added to a solution of compound 4-6 (1.9 g, 2.23 mmol) and intermediate B (1.39 g, 2.68 mmol) in dioxane (20 mL) and water (4 mL), and the mixture was stirred at 110°C for 16 h under nitrogen protection. The reaction solution was filtered through diatomaceous earth, and the filter cake was washed with 200 mL of ethyl acetate. The organic phase was collected, washed with water (150 mL) and saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, petroleum ether: ethyl acetate = 2:1) to obtain compound 4-7. LCMS (ESI) m/z: 1162.7 [M+H] ⁺ .

Step 7:

At 20°C, tetrabutylammonium fluoride (1M tetrahydrofuran solution, 6.12 mL) was added to a solution of compound 4-7 (1.78 g, 1.53 mmol) in tetrahydrofuran (30 mL), and the mixture was stirred at 20°C for 0.2 h. The reaction solution was diluted with 150 mL of ethyl acetate, washed with water (200 mL*3) and saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 4-8. LCMS (ESI) m/z: 1006.5 [M+H] ⁺ .

Step 8:

At 20°C, hydrogen chloride/ethyl acetate (4M solution, 17.69 mL) was added to a solution of compound 4-8 (1.53 g, 1.52 mmol) and ethyl acetate (15 mL), and the mixture was stirred at 20°C for 0.5 h. The reaction solution was filtered, the filter cake was collected, and the crude product was purified by preparative HPLC (chromatographic column: Phenomenex luna C18 250*80mm*10μm; mobile phase: [water (formic acid)-acetonitrile]; gradient: acetonitrile 25%-45%) to give the formate salt of compound 4-9A (LCMS retention time: 0.808 min), LCMS (ESI) m/z: 664.4 [M+H] ⁺ and the formate salt of compound 4-9B (LCMS retention time: 0.824 min), LCMS (ESI) m/z: 664.4 [M+H] ⁺ .

Step 9:

At 20°C, sodium L-ascorbate (83.57 mg, 421.83 μmol) and copper sulfate pentahydrate (105.32 mg, 421.83 μmol) were added to a solution of the formate salt of compound 4-9B (280 mg) and intermediate D (239.21 mg, 506.19 μmol) in tert-butyl alcohol (20 mL) and water (20 mL), and the mixture was stirred at 50°C for 16 h under nitrogen protection. Water (40 mL) was added to the reaction solution for dilution, and then extracted with methanol/ethyl acetate = 1/5 (40 mL*2), and the combined organic phase was washed with 10% ammonia water (100 mL*1) and saturated brine (100 mL*1), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by preparative HPLC (chromatographic column: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]: gradient 22%-52%), and further purified by preparative HPLC (column model: Waters Xbridge 150*25mm*5μm; mobile phase: [water (ammonium bicarbonate)-acetonitrile]: gradient acetonitrile 34%-54%) to obtain compound 4B. LCMS (ESI) m/z: 1136.7 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δppm 13.13 (br s, 1H), 8.98 (s, 1H), 8.64 (s, 1H), 8.49 (br d, J = 7.6Hz, 1H), 7.64 (br d, J = 8.0Hz, 2H), 7.53-7.35 (m, 7H), 6.74 (br d, J＝8.0Hz,2H),5.51-5.39(m,1H),5.32(br d,J＝10.0Hz,1H),5.19(br s,1H),5.05(br s,1H),4.95-4.82(m,2H),4.46(br t,J＝8.0Hz,1H),4.32(br s,1H),4 .24(br d,J＝7.6Hz,1H),3.83-3.73(m,3H),3.71-3.58(m,5H),3.33(br s,3H),3.11(br d,J＝9.6Hz,1H),3.04-2.88(m,2H),2.70(br dd,J＝13.2,5.6Hz,1H),2.5 3(br s,3H),2.46(s,3H),2.13-2.04(m,1H),1.98(br s,3H),1.90(br d,J＝9.2Hz,1H),1.84-1.70(m,2H),1.35(br d,J＝6.0Hz,4H),1.08(br d,J＝6.0Hz,3H ),0.72(br d,J＝6.0Hz,3H),0.67-0.50(m,4H).

Example 5

Synthesis route:

Step 1:

Under nitrogen protection, sodium hydrogen (826.89 mg, 20.67 mmol, purity: 60%) was added to a tetrahydrofuran (40 mL) solution of compound 4-5 (4 g, 6.89 mmol) and compound 5-1 (2.04 g, 8.27 mmol), and the resulting reaction solution was stirred at 40°C for 12 h. Water (50 mL) was added to the reaction solution, stirred for 5 min, and the aqueous phase was extracted with ethyl acetate (50 mL*2). The combined organic phase was washed with saturated brine (100 mL*3), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated, and the obtained crude product was purified by silica gel column chromatography (eluent, petroleum ether/ethyl acetate = 2/1) to obtain compound 5-2. LCMS (ESI) m/z: 806.3, 808.3 [M+H] ⁺ .

Step 2:

Under nitrogen protection, a mixed solution of compound 5-2 (2 g, 2.48 mmol), intermediate B (1.41 g, 2.73 mmol), potassium phosphate (1.58 g, 7.44 mmol) and tetrakis(triphenylphosphine)palladium (286.42 mg, 247.86 μmol) in dioxane (24 mL) and water (6 mL) was heated to 110 ° C and stirred for 12 h. Anhydrous sodium sulfate was added to the reaction solution for drying, filtration, and the filtrate was concentrated. The obtained crude product was purified by silica gel column chromatography (petroleum ether/ethyl acetate=2/1) to obtain compound 5-3. LCMS (ESI) m/z: 1118.6 [M+H] ⁺ .

Step 3:

Under nitrogen protection, tetrabutylammonium fluoride (2 mol/L tetrahydrofuran solution, 3 mL) was added to a solution of compound 5-3 (2.4 g, 2.15 mmol) in tetrahydrofuran (15 mL), and the resulting reaction solution was stirred at 20°C for 15 min. Water (20 mL) was added to the reaction solution, stirred for 5 min, and the aqueous phase was extracted with ethyl acetate (20 mL*2). The combined organic phase was washed with saturated brine (30 mL*3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was purified by silica gel column chromatography (eluent, petroleum ether/ethyl acetate = 2/1) to obtain compound 5-4. LCMS (ESI) m/z: 1000.4 [M+H] ⁺ .

Step 4:

Hydrogen chloride/ethyl acetate (2 mol/L, 20 mL) was added to a solution of compound 5-4 (2 g, 2.00 mmol) in ethyl acetate (5 mL), and the resulting reaction solution was stirred at 20° C. for 0.5 h. The reaction solution was concentrated, and the resulting crude product was purified by preparative HPLC (column model: Phenomenex luna C18 250*50 mm*15 μm; mobile phase A: water containing 0.1% trifluoroacetic acid; mobile phase B: acetonitrile; gradient: 20%-50% acetonitrile) to obtain trifluoroacetate salt of isomer 5-5A (retention time: 1.898 min, LCMS (ESI) m/z: 662.3 [M+H] ⁺ ) and trifluoroacetate salt of isomer 5-5B (retention time: 2.041 min, LCMS (ESI) m/z: 662.3 [M+H] ⁺ ).

Step 5:

At 25°C, 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (1.85g, 2.53mmol) and potassium acetate (7.45g, 75.90mmol) were added to a solution of compound D-1 (8g, 25.30mmol) and bis(triphenylphosphino)ferrocenepalladium chloride (1.85g, 2.53mmol) and potassium acetate (7.45g, 75.90mmol) in dioxane (100mL), and the mixture was stirred at 100°C for 2h under nitrogen protection. After the reaction, the reaction solution was cooled to room temperature, ethyl acetate (200mL) was added to the reaction solution, filtered through diatomaceous earth, the filter cake was washed with ethyl acetate (150mL*2), and the organic phase was concentrated under reduced pressure to obtain 5-6, which was directly used in the next step. LCMS (ESI) m/z: 308.1[M-56+1] ⁺ .

Step 6:

At 20°C, selenium powder (10.03 g, 126.98 mmol) was added to a solution of compound 5-7 (22 g, 126.98 mmol) in DMF (440 mL) and water (44 mL). The mixed solution was replaced with CO gas three times, and reacted under a CO atmosphere, heated to 90°C, and stirred for 5 h. After the reaction, the reaction solution was poured into water (900 mL), then extracted with ethyl acetate (500 mL*3), and the combined organic phases were washed with water (500 mL*3) and saturated brine (500 mL*1), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain product 5-8. LCMS (ESI) m/z: 256.0 [M+H] ⁺ .

Step 7:

At 20°C, 5-9 (10.92 g, 118.00 mmol) was added to a solution of compound 5-8 (30 g, 118.00 mmol) in ethanol (300 mL), and the mixture was stirred at 80°C for 1 h under nitrogen protection. After the reaction was completed, the reaction solution was cooled to 20°C, and the volatile components were removed by concentration under reduced pressure to obtain a crude product. Ethyl acetate (300 mL) was added to the crude product and stirred for 30 min, filtered, and the filter cake was washed with ethyl acetate (100 mL). The filter cake was added to a mixture of ethyl acetate (500 mL) and saturated sodium carbonate (500 mL), stirred for 30 min, the organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain product 5-10. LCMS (ESI) m/z: 294.1 [M+H] ⁺ .

Step 8:

At 20°C, selenium dioxide (7.97 g, 71.85 μmol) was added to a solution of compound 5-10 (21 g, 71.85 mmol) in dioxane (210 mL), and the mixture was stirred at 40°C for 16 h under nitrogen protection. After the reaction was completed, the reaction solution was cooled to room temperature, filtered, and the filtrate was poured into water (300 mL), then extracted with ethyl acetate (200 mL*3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, petroleum ether: ethyl acetate = 100:1) to obtain compound 5-11. LCMS (ESI) m/z: 308.0[M+H] ⁺ .

Step 9:

At 20°C, NBS (23.92 g, 134.40 mmol) was added to a DMF (200 mL) solution of compound 5-11 (19.6 g, 64.00 mmol), and the mixture was stirred at 50°C for 4 h under nitrogen protection. After the reaction, the reaction solution was cooled to room temperature, poured into water (500 mL), stirred for 30 min, filtered, and the filter cake was washed with water (200 mL), and concentrated to obtain compound 5-12. LCMS (ESI) m/z: 385.8 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d ₆ ) δppm 8.25-8.31 (m, 2H), 7.58-7.63 (m, 2H), 2.47 (s, 3H), 1.32 (s, 9H).

Step 10:

At 20°C, 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (1.85g, 2.53mmol) and sodium carbonate (6.70g, 63.25mmol) were added to a solution of compound 5-6 (9.19g, 25.30mmol) and 5-12 (9.74g, 25.30mmol) in dioxane (120mL) and water (24mL), and the mixture was stirred at 100°C for 2h under nitrogen protection. After the reaction was completed, 200mL of ethyl acetate was added to the reaction solution, filtered through diatomaceous earth, the filtrate was poured into water (200mL), and then extracted with ethyl acetate (150mL*2), the combined organic phase was washed with saturated brine (300mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, petroleum ether: ethyl acetate = 7:1 to 2:1) to obtain compound 5-13. LCMS(ESI)m/z: 543.1[M+H] ⁺ .

Step 11:

At 20°C, hydrogen chloride/ethyl acetate (4 mol/L, 100 mL) was added to a solution of compound 5-13 (13 g, 24.01 mmol) in ethyl acetate (50 mL), and the mixture was stirred at 20°C for 1 h. After the reaction was completed, the volatile components were removed under reduced pressure to obtain the hydrochloride of compound 5-14. LCMS (ESI) m/z: 443.1 [M+H] ⁺ .

Step 12:

At 20°C, sodium hydroxide (8.87 g, 221.81 mmol) was added to a solution of the hydrochloride salt (10.6 g) of compound 5-14 in ethanol (106 mL), the temperature was raised, and the mixture was stirred at 80°C for 5 h. After the reaction was completed, the reaction solution was cooled to room temperature, filtered, and the pH of the filtrate was adjusted to 10-11 with a hydrogen chloride/methanol solution (4 mol/L), filtered, the filtrate was collected, and the volatile components were removed under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC (column model: Kromasil Eternity XT 250*80mm*10μm; mobile phase: [water (ammonia water)-acetonitrile]: gradient 1%-30% acetonitrile) to obtain compound 5-15. LCMS (ESI) m/z: 283.1 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO-d ₆ ) δppm 9.94 (s, 1H), 7.41-7.45 (m, 2H), 7.36-7.39 (m, 2H), 3.90 (dd, J = 7.60, 4.88Hz, 1H), 3.47 (dd, J = 10.32, 4.88Hz, 1H), 3.33 (br d, J＝2.36Hz,1H),2.42(s,3H).

Step 13:

At 20°C, D-5 (2.82 g, 12.21 mmol), N,N-diisopropylethylamine (8.42 mL) and HATU (5.52 g, 14.51 mmol) were added to a DMF (34 mL) solution of compound 5-15 (3.4 g, 12.09 mmol), and the mixture was stirred at 20°C for 12 h under nitrogen protection. After the reaction, the reaction solution was poured into water (100 mL), then extracted with ethyl acetate (100 mL*4), and the combined organic phases were washed with saturated brine (150 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, methanol: ethyl acetate = 1:30 to 1:10) to obtain compound 5-16. LCMS (ESI) m/z: 496.2 [M+H] ⁺ .

Step 14:

At 20°C, hydrogen chloride/ethyl acetate solution (4 mol/L, 38 mL) was added to a solution of compound 5-16 (3.8 g, 7.69 mmol) in ethyl acetate (38 mL), and the mixture was stirred at 20°C for 0.5 h. After the reaction was completed, the volatile components were removed by distillation under reduced pressure to obtain the hydrochloride of 5-17. LCMS (ESI) m/z: 396.0 [M+H] ⁺ .

Step 15:

At 20°C, D-8 (1.68 g, 7.74 mmol), N,N-diisopropylethylamine (5.34 mL, 30.64 mmol) and HATU (3.50 g, 9.19 mmol) were added to a DMF (34 mL) solution of the hydrochloride salt of compound 5-17 (3.3 g), and the mixture was stirred at 20°C for 1 h under nitrogen protection. After the reaction was completed, the reaction solution was poured into water (100 mL), then extracted with ethyl acetate (100 mL*5), the combined organic phases were washed with saturated brine (150 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, petroleum ether: ethyl acetate = 0:1) to obtain compound 5-18. LCMS (ESI) m/z: 595.2 [M+H] ⁺ , 539.1 [M-56+H] ⁺ , 495.1[M-100+H] ⁺ .

Step 16:

At 20°C, hydrogen chloride/ethyl acetate solution (4 mol/L, 25 mL) was added to a solution of compound 5-18 (2.5 g, 4.21 mmol) in ethyl acetate (25 mL), and the mixture was stirred at 20°C for 1 h. After the reaction was completed, the volatile components were removed by distillation under reduced pressure to obtain the hydrochloride of 5-19. LCMS (ESI) m/z: 495.2 [M+H] ⁺ .

Step 17:

At 20°C, to a solution of the hydrochloride of compound 5-19 (178 mg), copper sulfate pentahydrate (26.84 mg, 107.49 μmol) and potassium carbonate (171.77 mg, 1.24 mmol) in methanol (10 mL), the hydrochloride of D-11 (189.03 mg, 901.80 μmol) was added, and the mixed liquid was stirred at 20°C for 16 h under a nitrogen atmosphere. After the reaction was completed, the solid phase was filtered off with diatomaceous earth, the filter cake was washed with ethyl acetate (20 mL), the liquid phase was collected and the volatile components were removed under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (silica gel, eluent, ethyl acetate: methanol = 30:1 to 20:1) to obtain compound 5-20. LCMS (ESI) m/z: 521.0 [M+H] ⁺ . ¹ H NMR (400MHz, CD ₃ OD) δppm 9.87-9.95(m,1H),7.43(s,2H),7.18(s,2H),7.03-7.16(m,1H),5.02(br t,J＝6.04Hz,1H),4.68(t,J＝8.36Hz,1H),4.47(br s ,1H),3.79-3.95(m,3H),3.65-3.76(m,3H),2.45(s,3H),1.91-2.33(m,4H),0.99-1.08(m,6H).

Step 18:

Under nitrogen protection, compound 5-20 (122.24 mg, 235.32 μmol), sodium ascorbate (42.38 mg, 213.92 μmol) and copper sulfate pentahydrate (53.41 mg, 213.92 μmol) were added to a mixed solution of trifluoroacetate salt of compound 5-5B (0.2 g) in tert-butyl alcohol (20 mL) and water (20 mL), and the reaction solution was stirred at 50° C. for 12 h. 10% aqueous ammonia (50 mL) was added to the reaction solution at 20° C. and stirred for 10 min, and extracted with a mixed solvent of ethyl acetate/methanol = 7/1 (50 mL*2). The combined organic phases were washed with saturated brine (100 mL*3), dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and the crude product was purified by preparative HPLC (column model: Phenomenex luna C18 150*25 mm*10 μm; mobile phase A: water containing 0.01% trifluoroacetic acid; mobile phase B: acetonitrile; gradient: 19%-49% acetonitrile) to obtain the trifluoroacetate salt of compound 5B. ¹ H NMR (400MHz, DMSO-d ₆ ) δ9.97 (s, 1H), 9.37-9.18 (m, 1H), 8.66 (s, 1H), 8.54-8.37 (m, 2H), 7.66 (d, J = 8.1Hz, 2H), 7.48-7.33 (m, 8H), 6.73 (br d, J = 8.3Hz, 2H),5.61-5.45(m,1H),5.34(d,J＝10.1Hz,1H),5.25(br s,1H),5.23-5.18(m,1H),4.91-4.83(m,1H),4.77-4.69(m,1H),4.57-4.50(m,2H),4.47-4.43(m,1H),4.33(br d,J＝2.0Hz,1H),4.10(br s,1H),3.7 7-3.77(m,1H),3.72(br s,1H),3.62(br d,J＝2.4Hz,2H),3.44-3.40(m,1H),3.36(s,3H),3.02-2.92(m,1H),2.75(br dd,J＝5.5,13.3Hz,1H),2.56(s,3H),2.44(s,3H),2.30(br d,J＝11.5Hz,1H),2.13-2.04(m,2H),2.00(br d,J＝2.4Hz,4H),1.86-1.73(m,1H),1.40(br d, J＝6.1Hz,3H),1.16-1.03(m,4H),1.02-0.90(m,1H),0.77-0.58(m,8H); LCMS(ESI)m/z:1182.7[M+H] ⁺ .

Biological activity evaluation

Experimental Example 1: In vitro cell proliferation assay

Experimental Materials:

RPMI-1640 medium, DMEM medium, penicillin/streptomycin antibiotics were purchased from Gibco, and fetal bovine serum was purchased from Hyclone. 3DCellTiter-Glo (cell viability chemiluminescent detection reagent) reagent was purchased from Promega. GP2D cell line (DMEM + 10% FBS + 1% penicillin/streptomycin) was purchased from ECACC, and PK-59 cell line (DMEM + 10% FBS + 1% penicillin/streptomycin) was purchased from Nanjing Kebai Biotechnology Co., Ltd. A375 cell line (DMEM + 10% FBS + 1% penicillin/streptomycin) was purchased from ATCC, Envision multi-label analyzer (PerkinElmer).

Experimental methods:

The cells were seeded in an ultra-low attachment 96-well U-shaped plate, with 80 μL of cell suspension per well, containing 1000 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture.

The compound to be tested was diluted 5-fold into 8 concentrations, i.e. from 2 mM to 25.6 nM, using a pipette, and a double-well experiment was set up. Add 78 μL of culture medium, and then transfer 2 μL of the gradient dilution compound per well to the middle plate according to the corresponding position, mix well and transfer 20 μL per well to the cell plate. The concentration range of the compound transferred to the cell plate is 10 μM to 0.128 nM. The cell plate is placed in a carbon dioxide incubator and cultured for 5 days. Prepare another cell plate and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis.

Add 100 μL of cell viability chemiluminescent detection reagent to the cell plate and incubate at room temperature for 30 min to stabilize the luminescent signal. Read using a multi-label analyzer.

Data Analysis:

The raw data were converted into inhibition rate using the equation (Sample-Min)/(Max-Min)*100%, and the _IC50 value was obtained by four-parameter curve fitting (obtained in the "log (inhibitor) vs. response--Variable slope" mode in GraphPad Prism). The results of the inhibitory activity test of the compounds of the present invention on cell proliferation are shown in Table 2.

Table 2: Results of in vitro cell proliferation inhibition activity test of the compounds of the present invention

Experimental conclusion: The compounds of the present invention exhibit excellent in vitro activity against G12D mutated cells, but have no inhibitory activity against A375 cells with low G12D expression.

Experimental Example 2: In vitro AsPC-1 KRAS G12D degradation detection

Experimental Materials:

RPMI-1640 medium was purchased from GIbco, penicillin/streptomycin antibiotics were purchased from Vicente, and fetal bovine serum was purchased from Biosera. Protein primary and secondary antibodies were purchased from Cell Signaling Technology. AsPC-1 cell line was purchased from ATCC, and 4% paraformaldehyde, Triton-100, etc. were purchased from Bio-Tech. Envision multi-label analyzer (PerkinElmer).

Experimental methods:

The cells were seeded in a 96-well black-walled plate, with 80 μL of cell suspension per well, containing 50,000 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture.

The compound to be tested was diluted 5-fold to 8 concentrations using a gun, i.e., from 2mM to 25.6nM, and a double-well experiment was set up. 78μL of culture medium was added to the middle plate, and then 2μL of the gradient dilution compound per well was transferred to the middle plate according to the corresponding position, and 20μL of each well was transferred to the cell plate after mixing. The concentration range of the compound transferred to the cell plate was 10μM to 0.128nM. After the cell plate was placed in a carbon dioxide incubator for 24h, the cells were removed, the culture medium was discarded, and the cells were fixed with 4% PFA fixative for 30min, washed 3 times with PBS, permeabilized with 0.1% Triton for 20min, washed 3 times with PBS, blocked with 0.1% BSA solution at room temperature for 1h, incubated with protein primary antibody at 4℃ overnight, washed 3 times with PBST, incubated with fluorescent secondary antibody (FITC or Cy5 labeled) at room temperature for 2h, and washed 3 times with PBST.

Data Analysis:

Calculate the FITC/Cy5 in the sample well based on the measured fluorescence intensity

FITC/Cy5=(FITCsample-FITCblank)/(FITCsample-FITCblank)

The inhibition % under compound treatment was calculated as [1-(FITC/Cy5comp.)/(FITC/Cy5DMSO)]*100, and the DC ₅₀ value was obtained by four-parameter curve fitting (obtained by "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism). The results are shown in Table 3.

Experimental Example 3: Detection of p-ERK level in AsPC-1 cells

Experimental Materials:

AsPC-1 cells were purchased from ATCC; RPMI-1640 medium was purchased from GIbco; fetal bovine serum was purchased from Hyclone; Advanced Phospho- ERK1/2 (THR202/TYR204) KIT was purchased from Bioauxilium, see Table 3 for details.

Table 3: Advanced Phospho-ERK1/2 (THR202/TYR204) KIT ingredients

Experimental methods:

1. Plant AsPC-1 cells in a 384-well cell culture plate with a white bottom, 8 μL of cell suspension per well, each well contains 7500 cells, and place the cell plate in a carbon dioxide incubator and incubate at 37 degrees overnight;

2. Dilute the compound to be tested with 100% DMSO to 3mM as the first concentration, and then use a pipette to dilute it into ten concentrations of 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1μM. Take 2μL of compound and add it to 198μL of cell starvation medium. After mixing, take 15μL of compound solution and add it to 35μL of cell starvation medium and mix. Then, add 4μL of the compound solution in the last step to the corresponding cell plate wells per well. Put the cell plate back into the carbon dioxide incubator and continue to incubate for 3h. At this time, the compound concentrations are 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1nM.

3. After the incubation, add 3 μL 5X cell lysis buffer to each well and incubate at room temperature for 30 minutes with shaking;

4. Use detection buffer to dilute Phospho-ERK1/2 Eu Cryptate antibody and Phospho-ERK1/2 d2 antibody 20 times, mix them in a 1:1 ratio, add 5 μL to each well of the cell culture plate, and incubate at room temperature for 2 hours.

5. After incubation, use a multi-label analyzer to read HTRF excitation: 320nm, emission: 615nm, 665nm.

Data Analysis:

The raw data were converted into inhibition rate using the equation (Sample-Min)/(Max-Min)*100%, and the _IC50 value was obtained by four-parameter curve fitting (obtained by log(inhibitor) vs.response--Variable slope mode in GraphPad Prism). The results of the inhibition test of the compounds of the present invention on p-ERK are shown in Table 4.

Max well: The reading value of the positive control well is 1X lysate

Min well: negative control well reading value is 0.5% DMSO cell well cell lysate

Table 4: Results of in vitro screening test of compounds of the present invention

Experimental conclusion: The compounds of the present invention exhibit excellent G12D degradation activity and have good inhibitory activity on downstream pERK.

Experimental Example 4: Antitumor Effect on Human Pancreatic Cancer PK-59 Xenograft Tumor Model

Female Balb/c nude mice were inoculated subcutaneously with PK-59 human pancreatic cancer cells and randomly divided into groups (6 animals per group) according to tumor volume and body weight on day 6 after inoculation, and the drugs were administered as described below.

Vehicle control group: drug administration began in the afternoon of the day of grouping (the day of drug administration was the drug administration D0 day), and the solvent (ethanol, 5% glucose solution, 1M hydrochloric acid solution, 50% (2-hydroxypropyl)-β-cyclodextrin (HP-βCD) aqueous solution, HCO-40, 1M sodium hydroxide aqueous solution = 4:84.4:1.1:1:9:0.5) was injected into the tail vein at a dose of 0.1 mL/10 g body weight once a week.

Compound administration group: Administration began in the afternoon of the day of grouping, and was administered once a week by tail vein injection at a dose of 0.1 mL/10 g body weight. During the experiment, the tumor volume was measured twice a week and the mice were weighed. The tumor volume was calculated according to the formula of length × width 2/2. The tumor proliferation rate was calculated according to the formula: tumor proliferation rate = tumor volume of treatment group/tumor volume of control group × 100%. The Student’s t-test was used for statistical analysis between the groups, and p<0.05 was considered to be significantly different. The results of the in vivo efficacy study are shown in Table 5.

Table 5: In vivo efficacy study results

Experimental conclusion: In the human pancreatic cancer PK-59 xenograft tumor model, the compound of the present invention exhibits good efficacy and safety and has a good tumor inhibition effect in vivo.

Claims (18)

A compound of formula (VI), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,
in, T is selected from S and Se; R ₁ is selected from H, D and C _1-3 alkyl; _R2 is selected from H, D and _C1-3 alkyl; R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl; R ₅ is selected from -OR ₄ ; R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _a ; each R ₆ and R ₇ is independently selected from H, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ and CH ₂ F; Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl )-( _OC1-3alkyl ), wherein said -C(=O) _C1-3alkyl , -N( _C1-3alkyl )-( _OC1-3alkyl ) are independently optionally substituted by 1, 2 or 3 R; Each R _b is independently selected from D, F, Cl, Br, I, OH, NH ₂ and CH ₃ ; Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH; m is selected from 0, 1, 2, 3 and 4; Provided that: when T is selected from S, _R1 and _R2 are both H, _R3 is not H and F, and m is not 0. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

in, R ₁ is selected from H, D and C _1-3 alkyl; _R2 is selected from H, D and _C1-3 alkyl; R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl; R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R _a ; R ₅ is selected from -O-4-6 membered heterocycloalkyl and -OC _1-3 alkyl-N(C _1-3 alkyl)(OC _1-3 alkyl), wherein the C _1-3 alkyl is each independently optionally substituted by 1, 2 or 3 R; each R ₆ and R ₇ is independently selected from H, F, Cl, Br, I, CH ₃ , CF ₃ , CHF ₂ and CH ₂ F; Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl , wherein said -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl are independently optionally substituted by 1, 2 or 3 R; Each R _b is independently selected from D, F, Cl, Br, I, OH, NH ₂ and CH ₃ ; Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH; m is selected from 0, 1, 2, 3 and 4; Provided that: when _R1 and _R2 are both H, _R3 is not H and F, and m is not 0. The compound according to claim 2, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

in, R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 R _a ; R ₅ is selected from -O-4-6 membered heterocycloalkyl and -OC _1-3 alkyl-N(C _1-3 alkyl)(OC _1-3 alkyl); R ₃ , _Ra and m are as defined in claim 1 . The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from H and D. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₂ is selected from H, D and CH ₃ . The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₃ is selected from H and Cl. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein each _Ra is independently selected from -C(=O) _CH3 and -N( _CH3 ) _-OCH3 . The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₄ is selected from propyl, isopropyl, tetrahydropyranyl and pyrrolidinyl, and the propyl, isopropyl, tetrahydropyranyl and pyrrolidinyl are each independently optionally substituted by 1, 2 or 3 _Ra . The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₄ is selected from Said are each independently optionally substituted with 1, 2 or 3 _Ra . The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₄ is selected from The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein R ₅ is selected from The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:
in, R ₄ is selected from C _1-6 alkyl and 4-7 membered heterocycloalkyl, wherein the C _1-6 alkyl and 4-7 membered heterocycloalkyl are optionally substituted by 1, 2 or 3 R _a ; R ₅ is selected from -O-4-6 membered heterocycloalkyl and -OC _1-3 alkyl-N(C _1-3 alkyl)(OC _1-3 alkyl), wherein the C _1-3 alkyl is each independently optionally substituted by 1, 2 or 3 R; R ₃ is selected from H, D, F, Cl, Br, CN, NH ₂ , OH and C _1-3 alkyl; Each _Ra is independently selected from -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl , wherein said -C(=O) _C1-3alkyl and -N( _C1-3alkyl ) _-OC1-3alkyl are independently optionally substituted by 1, 2 or 3 R; Each R is independently selected from D, F, Cl, Br, I, =O, CN, _NH2 and OH; m is selected from 0, 1 and 2. The following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:

The compound according to claim 13, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

The compound according to claim 14, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:


Use of the compound according to any one of claims 1 to 15, its stereoisomer or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating a disease associated with a KRAS mutant protein. The use according to claim 16 is characterized in that the KRAS mutant protein-related disease is selected from solid tumors with KRAS G12D mutation. The use according to claim 17 is characterized in that the solid tumor with KRAS G12D mutation is selected from pancreatic cancer, lung cancer or colon cancer.