CN118580238A - Pyrroloheterocyclic compounds and preparation methods and applications thereof - Google Patents

Abstract

The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a class of pyrrolo-heterocyclic compounds. The compound has better KRAS G12V mutant gene inhibition effect and can be used for tumor diseases related to KRAS G12V gene mutation.

Description

Pyrrolo-heterocyclic compound, and preparation method and application thereof

Technical Field

The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to a class of pyrrolo-heterocyclic compounds. The compound has better KRAS G12V mutant gene inhibition effect and can be used for KRAS G12V gene mutation-mediated tumor diseases.

Background

The RAS gene is one of the most common mutant genes in cancer cells, with RAS gene mutation detected in 30% of human tumors. The RAS subfamily comprises HRAS, NRAS, KRAS, about 85% of most common in cancer cells with KRAS mutations. KRAS mutations and/or KRAS wild-type amplifications are common to colorectal cancer (about 45% in the united states, 49% in china) and pancreatic cancer (about 90% in the united states, about 87% in china) and non-small cell lung cancer (subtype adenocarcinomas: about 35% in the united states, about 13% in china). KRAS mutations have also been found in some cancers, including gastric cancer, uterine cancer, cervical cancer, bladder cancer, cholangiocarcinoma, diffuse large B-cell lymphoma, multiple myeloma, cutaneous squamous cell carcinoma, and the like. KRAS allele distribution varies with tumor type, with G12D (41%), G12C (14%) and G12V (28%) being most common, G12C mutating 13.6% of all lung adenocarcinomas, while G12D and G12V are the two most common alleles in colorectal and pancreatic cancers.

KRAS proteins are a class of guanine nucleotide binding proteins with GTP hydrolase activity that act as molecular switches in a range of intracellular signaling pathways. The balance of nucleotide hydrolysis and exchange determines the level of KRAS in the intracellular activation state. When bound to GDP, KRAS is in an "OFF" state, and GTP is exchanged with GDP by a guanylate exchange factor (e.g., SOS 1) to put KRAS in an "ON" state. In this state, KRAS may activate downstream signaling pathways, including mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3 kinase (PI 3K) pathways, thereby promoting cell proliferation and survival. In cancer cells, mutations in KRAS disrupt guanine-promoted exchange processes, typically by becoming GAP-independent and "locking" the RAS in an active GTP-bound state, thereby activating downstream signaling pathways that lead to tumor cell growth.

In one aspect, the affinity of KRAS for GTP is picomolar, whereas the concentration of GTP in the cell is as high as 0.5 μm. Thus, it is difficult for small molecules to compete effectively with GTP. On the other hand, KRAS proteins have a nearly spherical structure, with no deep hydrophobic pocket, nor obvious binding sites, due to their smoother surface. Targeting KRAS has therefore been considered very challenging in the last decades. Candidate compounds targeting KRAS G12C have been developed in recent years, and Sotorasib (AMG 510) of the incorporation company was FDA approved for marketing in month 5 of 2021 as the first global drug targeting KRAS G12C. However, there is no candidate molecule on the market for other mutations, so developing KRAS inhibitors against other subtype mutations as soon as possible and pushing them to the clinical stage is also an important development area at present.

Disclosure of Invention

In a first aspect of the present invention, there is provided a compound of formula I or a pharmaceutically acceptable salt thereof, a stereoisomer thereof;

Wherein ring B is a 5-6 membered heteroaryl;

Ring A and ring C are fused together to form a 7-12 membered heterobicyclic group, wherein ring A is a 5-6 membered heteroaromatic ring and ring C is selected from the group consisting of benzene ring, 5-6 membered heteroaromatic ring, and 5-6 membered heterocyclic ring;

Ra is independently selected from halogen, C _1-4 alkyl, C _1-4 alkyl-S (O) ₂ -, 3-10 membered heterocyclyl -(CR¹R²)q₁-X¹-、N(R³)(R⁴)-(CR⁵R⁶)q₂-X²-; wherein said heterocyclyl is optionally substituted with one or more of: halogen, hydroxy, C _1-4 alkyl, C _1-4 alkoxy-C _1-4 alkyl-, -N (R ⁷)(R⁸), optionally substituted 3-6 membered heterocyclyl;

X ¹、X² is independently selected from a chemical bond and O;

q ₁、q₂ are each independently selected from 0,1,2, 3;

R ¹、R² is independently selected from H, C _1-4 alkyl;

R ³、R⁴ is independently selected from H, C _1-4 alkyl, or R ³、R⁴ forms a 4-6 membered heterocyclic ring with the N atom to which it is attached;

r ⁵、R⁶ is independently selected from H, C _1-4 alkyl, or R ⁵、R⁶ on the same C atom forms an optionally substituted 3-6 membered cycloalkyl group with the C atom to which it is attached;

R ⁷、R⁸ is independently selected from H, C _1-4 alkyl;

Rc is independently selected from H, C _1-4 alkyl, CN-C _1-4 alkyl, - (CR ⁹R¹⁰)s-Cy、N(R¹¹)(R¹²)-C_1-4 alkyl-;

cy is selected from the following groups optionally substituted with 1-3 Rx: 5-10 membered heteroaryl, phenyl, 3-6 membered heterocyclyl, 3-6 membered cycloalkyl, 5-6 membered cycloalkenyl;

Rx is selected from oxo, hydroxy, halogen, C _1-4 alkyl, -N (R ¹³)(R¹⁴);

s is selected from 0, 1, 2, 3;

R ⁹、R¹⁰ is independently selected from H, C _1-4 alkyl, halogen, halogenated C _1-4 alkyl, C _3-6 cycloalkyl, or R ⁹、R¹⁰ on the same carbon atom together with the C atom to which it is attached form a 3-6 membered cycloalkyl;

R ¹¹、R¹²、R¹³、R¹⁴ is independently selected from H, C _1-4 alkyl, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl;

m and n are each independently selected from 0,1,2, and 3.

In one embodiment of the invention, ring B is selected from the following groups:

* Indicating that this position is connected to ring a.

In one embodiment of the invention, ring B is selected from the following groups: * Indicating that this position is connected to ring a.

In one embodiment of the invention, ring A is selected from a pyridine ring or a pyrimidine ring.

In one embodiment of the invention, ring C is selected from the group consisting of pyrrole rings, benzene rings, pyridine rings, pyridazine rings, imidazole rings, tetrahydropyrrole rings, piperidine rings, morpholine rings, piperazine rings, 1, 4-oxazaalkanes.

In one embodiment of the invention, ring a is selected from a pyridine ring or a pyrimidine ring; ring C is selected from pyrrole ring, benzene ring, pyridine ring, pyridazine ring, imidazole ring, tetrahydropyrrole ring, piperidine ring, morpholine ring, piperazine ring, 1, 4-oxazaane.

In one embodiment of the invention, ra is independently selected from 5-6 membered heterocyclyl-CR ¹R² -O-, wherein the heterocyclyl is optionally substituted with 1-3 groups selected from halogen, C _1-4 alkyl; r ¹ is selected from H; r ² is selected from C _1-4 alkyl.

In one embodiment of the invention, ra is selected from the following groups: cl, F, -S (O) ₂CH₃,

In one embodiment of the invention, rc is independently selected from-CR ⁹R¹⁰-Cy,R⁹ and R ¹⁰ is independently selected from H, F, cl, -CH ₃、-CF₃, cyclopropyl, or R ⁹、R¹⁰ together with the C atom to which it is attached forms cyclopropyl;

cy is selected from the following groups optionally substituted with 1-3 Rx: pyridyl, pyrazolyl, 3-6 membered heterocyclyl, 3-6 membered heterocycloalkyl;

Rx is selected from oxo, hydroxy, halogen, C _1-4 alkyl, -N (R ¹³)(R¹⁴);

R ¹³、R¹⁴ is independently selected from H, C _1-4 alkyl, deuterated C _1-4 alkyl, halogenated C _1-4 alkyl.

In one embodiment of the invention, rc is selected from H, -CH ₃、-CH(CH₃)₂、CNCH₂CH₂ -, cyclopentyl

In one embodiment of the invention, the 7-12 membered heterobicyclic group to which ring A and ring C are co-fused is selected from the following rings:

In one embodiment of the invention, the structural unit Selected from the group consisting of

Wherein m, n are each independently selected from 0, 1, 2, 3, ra, rc are as defined above

In one embodiment of the invention, the structural unitSelected from the group consisting of

Wherein Ra, rc are as defined above. In one aspect of the present invention,Selected from the following structures:

in one embodiment of the present invention, the compound of formula I is preferably a compound having the structure:

Wherein Ra, rc are as defined above;

Z ¹、Z² is independently selected from CH, N, and at least one is selected from N.

In one embodiment of the invention of formula Id or formula Id-1, Z1 and Z2 are both N.

In one embodiment of the invention of formula Id or formula Id-1, ra is 3-10 membered heterocyclyl-CR ¹R² -O-; the R ₁、R₂ is independently selected from H, methyl, ethyl, isopropyl, trifluoromethyl and trichloromethyl; the 3-10 membered heterocyclyl is optionally substituted with 1 or more of the following: halogen, hydroxy, C _1-4 alkyl, C _1-4 alkoxy-C _1-4 alkyl-, -NH ₂, optionally substituted 3-6 membered heterocyclyl; preferably, ra is 5 membered heterocyclyl-CH (R ₂) -O-; the 5 membered heterocyclic ring may be optionally substituted with halogen, hydroxy, C _1-4 alkyl.

In one embodiment of the invention of formula Id or formula Id-1, ra is selected from

In one embodiment of the invention of formula Id or formula Id-1, ra is

In one embodiment of formula Id or formula Id-1 of the present invention, rc is independently selected from H, C _1-4 alkyl, - (CR ⁹R¹⁰) -Cy;

Cy is selected from the following groups optionally substituted with 1-3 Rx: 5-10 membered heteroaryl, phenyl; rx is selected from oxo, hydroxy, halo, C _1-4 alkyl, -NH ₂;R⁹、R¹⁰ are each independently selected from H, C _1-4 alkyl, halo C _1-4 alkyl, C _3-6 cycloalkyl;

in one embodiment of formula Id or formula Id-1 of the present invention, rc is 1 or more Rx substituted pyridinyl;

in one embodiment of formula Id or formula Id-1 of the present invention, rc is 1 or more Rx substituted pyridinyl;

In one embodiment of formula Id or formula Id-1 of the present invention, rc is selected from methyl; 1- (2-aminopyridin-3-yl) -ethyl;

in one embodiment of the invention, formula Id or formula Id-1 has the structure shown as formula Id-2 or formula Id-3 below;

Or alternatively

Wherein R ₁₀ is selected from H, C _1-4 alkyl, halogen, halogenated C _1-4 alkyl, C _3-6 cycloalkyl; rx is selected from hydroxy, halogen, C _1-4 alkyl, -NH ₂;

in one embodiment of formula Id-2 or formula Id-3 of the present invention, R ₁₀ is selected from H, methyl, trifluoromethyl;

In one embodiment of formula Id-2 or formula Id-3 of the present invention, rx is selected from methyl, F, NH ₂;

In one embodiment of formula Id-2 or formula Id-3 of the present invention, rx is selected from NH ₂;

In one embodiment of formula Id-2 or formula Id-3 of the present invention, R ₁₀ is selected from H or methyl, rx is NH ₂;

In one embodiment of the present invention, the following compounds or pharmaceutically acceptable salts thereof are preferred:

In one embodiment of the present invention, the following compounds or pharmaceutically acceptable salts thereof are preferred:

In one embodiment of the present invention, the compounds of the present invention are preferably the following compounds or pharmaceutically acceptable salts thereof:

In one embodiment of the present invention, the compounds of the present invention are preferably the following compounds or pharmaceutically acceptable salts thereof:

in a second aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, and any pharmaceutically acceptable carrier.

In a third aspect, the invention provides the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the manufacture of a medicament for the treatment of a disease associated with a KRAS G12V gene mutation.

In a fourth aspect, the invention provides the use of a pharmaceutical composition according to the second aspect of the invention in the manufacture of a medicament for the treatment of a disease associated with a KRAS G12V gene mutation.

In a fifth aspect, the invention provides a method of treating a disease associated with a KRAS G12V gene mutation, comprising administering to a patient in need thereof an effective amount of a compound of formula I according to the first aspect of the invention or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or a pharmaceutical composition according to the second aspect of the invention.

In the present invention, the disease associated with KRAS G12V gene mutation may be lung cancer associated with KRAS G12V gene mutation.

Definition and description

In the present invention, in particular in the compounds of formula I and their lower sub-formulae, said H refers to a hydrogen element, i.e. an atom whose nucleus has 1 proton, comprising three isotopes of protium (P), deuterium (D) and tritium (T);

in the present invention, the term "independently selected" means that the same or different ranges may be selected between different variable groups or when the same substituent is at different substitution positions. For example, the structural units of the present invention When n is 2 or more, each Ra group is independent of each other and may be the same or different; similarly, when m is 2 or more, each Rc group may be the same or different independently of each other.

"Oxo" as used herein means that the group is replaced by a "=o" structure, such as "-CH ₂", "=ch", "is oxo to" -C (O) - "," S "is oxo to" S (O) "or" S (O) ₂ ".

In the invention, in the 3-10 membered heterocyclic group -(CR¹R²)q₁-X¹-、N(R³)(R⁴)-(CR⁵R⁶)q₂-X²-, when q ₁ or q ₂ is 0, the CR ¹R² group is not present, and the groups on the left and right sides are directly connected through chemical bonds, namely the 3-10 membered heterocyclic group-X ¹-、N(R³)(R⁴)-X² -. Similarly, in- (CR ⁹R¹⁰) s-Cy, when s is 0, the group is Cy-.

In the present invention, the "halogen" or "halogen atom" means fluorine, chlorine, bromine, iodine; by "halo" is meant a group formed upon substitution of one or more hydrogen atoms in a substituent with a halogen atom.

The term "alkyl" refers to a straight or branched hydrocarbon group that is singly linked between carbon atoms, and between carbon atoms and hydrogen atoms. Alkyl is preferably C _1-4 or C _1-6 alkyl; c _1-4 alkyl "represents a straight or branched alkyl group having 1 to 4 carbon atoms; specific examples of C1-4 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, preferably methyl, ethyl, n-propyl, isopropyl; "C _1-6 alkyl" means a straight or branched chain alkyl group having 1 to 6 carbon atoms. Examples of C1-6 alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl.

As used herein, "CN-C _1-4 alkyl" refers to C _1-4 alkyl substituted with one or more cyano groups, specific examples include, but are not limited to: And the like, more preferably a C _1-4 alkyl group substituted with 1 CN.

The term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced with halogen atoms. The haloalkyl is preferably a C _1-4 haloalkyl, a C _1-6 haloalkyl. Examples of C _1-4 haloalkyl include, but are not limited to, monofluoromethyl, chloromethane, difluoromethyl, dichloromethane, trifluoromethyl, trichloromethyl, tribromomethyl.

The term "alkoxy" refers to alkyl-O-, alkyl as previously described, including C _1-4 alkoxy or C _1-6 alkoxy; specific examples include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, neopentoxy, n-hexyloxy.

The term "haloalkoxy" refers to an alkoxy group in which one or more hydrogen atoms are replaced with halogen atoms. Examples of C _1-4 haloalkoxy groups include, but are not limited to, trifluoromethoxy, trichloromethoxy, 2-trifluoroethoxy, 2-trichloroethoxy.

The term "cycloalkyl" refers to a cyclic saturated monocyclic or polycyclic hydrocarbon group formed by single bond attachment between carbon atoms. The ring C atom of which is optionally oxo, i.e., -CH ₂ -is oxo to form-C (O) -. The polycyclic hydrocarbon group comprises a spiro ring group, a bridged ring group and a condensed ring group. The "spirocyclic group" refers to a cyclic structure formed by sharing one carbon atom between two adjacent rings; the "bridged ring radical" refers to a ring structure formed by two non-adjacent carbon atoms shared by two adjacent rings; the term "fused ring group" refers to a cyclic structure formed by two adjacent carbon atoms shared between two adjacent rings; specifically included are "C _3-10 alkyl", "C _3-6 alkyl", "C _5-6 alkyl", "C _6-8 alkyl", "C _7-8 bridged cycloalkyl", "7-11 membered spirocycloalkyl", "7-10 membered fused ring alkyl", wherein the numerical values represent the number of ring carbon atoms. Specific examples include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,Etc.

The term "heterocyclyl" or "heterocycloalkyl" refers to monocyclic and polycyclic groups derived from the replacement of one or more ring carbon atoms in the cycloalkyl group with one or more heteroatoms or groups of heteroatoms; the heteroatom or heteroatom group is generally selected from N, N (O), O, S, S (O), S (O) ₂; the ring C atom of which is optionally oxo, i.e., -CH ₂ -is oxo to form-C (O) -. Cycloalkyl is as defined above. The polycyclic heterocyclic group includes a condensed heterocyclic group, a spiro heterocyclic group, and a bridged heterocyclic group. The heterocyclic group preferably contains 1 to 2 heteroatoms selected from N and/or O, more preferably 1N and 0 to 1N or O heteroatoms; the heterocyclic group is preferably a "nitrogen-containing heterocyclic group", which means a heterocyclic ring in which at least one ring atom is an N atom, and which may contain other hetero atoms; the heterocyclic group includes "3-11 membered heterocyclic group", "4-9 membered heterocyclic group", "7-9 membered condensed heterocyclic group", "7-8 membered bridged heterocyclic group", "7-9 membered spiro heterocyclic group"; the heterocyclic group is preferably a 3-6-membered, 3-7-membered, 4-6-membered, 4-7-membered, 5-6-membered, 5-7-membered heterocyclic group. the 3-membered heterocycloalkyl group includes oxiranyl and aziridinyl; the 4-membered heterocycloalkyl group includes oxetanyl, azetidinyl, thietane; the 5-membered heterocycloalkyl group includes an oxo-cyclopentyl group, an azacyclopentyl group, a thiacyclopentyl group; 6 membered heterocycloalkyl includes piperidinyl, piperazinyl, morpholinyl, 1,4 dioxane. Specific examples of the spiroheterocyclyl group include, but are not limited to: spiro [2.2] pentyl, spiro [2.3] hexyl, spiro [2.4] heptyl, spiro [2.5] octyl, spiro [2.6] nonyl, spiro [3.3] heptyl, spiro [3.4] octyl, spiro [3.5] nonyl, spiro [3.6] decyl, spiro [4.4] nonyl, spiro [4.5] decyl, spiro [4.6] undecyl, spiro [5.5] undecyl, spiro [5.6] dodecyl, spiro [6.6] tridecyl, spiro [6.7] tetradecyl. Examples of heterospirocyclic groups also include 2-oxa-7-azaspiro [5.3] nonan-7-yl, 2-oxa-6-azaspiro [3.3] heptan-6-yl, 2, 6-diazaspiro [3.3] heptan-2-yl, 2, 7-diazaspiro [5.3] nonyl, 2, 7-dioxaspiro [5.3] nonyl, 3, 9-diazaspiro [5.5] undec-3-yl, 1-oxa-4, 9-diazaspiro [5.5] undec-9-yl, 1-oxa-4, 8-diazaspiro [5.4] decane-8-yl, 3-azaspiro [5.5] undec-3-yl, 7-azaspiro [3.5] decane-7-yl, 1-oxa-4, 9-diazaspiro [5.5] undec-4-yl, 6-oxa-2, 9-diazaspiro [4.5] decan-9-yl, 9-oxa-2, 6-diazaspiro [4.5] decan-6-yl, 3-azaspiro [5.5] undec-3-yl, 4-oxa-1, 9-diazaspiro [5.5] undec-9-yl. Specific examples of the bridged heterocyclic group include, but are not limited to: bicyclo [3.1.0] hexyl, bicyclo [3.2.0] heptyl, bicyclo [3.3.0] octyl, bicyclo [4.1.0] heptyl, bicyclo [4.2.0] octyl, bicyclo [4.3.0] nonyl, bicyclo [4.4.0] decyl, and bicyclo [3.2.1] octyl. Examples of heterobridged ring radicals include, but are not limited to, 1, 4-diazabicyclo [4.4.0] decan-4-yl, 1, 4-diazabicyclo [4.3.0] -nonan-4-yl, 8-oxa-1, 4-diazabicyclo [4.4.0] decan-4-yl, 1, 4-diazabicyclo [4.4.0] decane-4-yl, 4, 7-diazabicyclo [4.3.0] nonan-4-yl, 2-oxa-5-azabicyclo [2.2.1] heptan-5-yl, 3, 7-diazabicyclo [4.3.0] nonan-3-yl, 3, 7-diazabicyclo [3.3.0] octan-3-yl, 3, 7-diazabicyclo [4.4.0] decan-3-yl, 3, 6-diazabicyclo [4.3.0] nonan-3-yl, 3, 6-diazabicyclo [4.4.0] decan-3-yl, 3,6, 9-triazabicyclo [4.4.0] decan-3-yl.

The invention is particularly described when the building blockIs that Or other similar structures, ring A is pyrimidinyl and ring C is considered heterocyclyl, i.e., tetrahydropyrrole, morpholinyl, 1, 4-oxazacyclyl.

The term "aryl" refers to an unsaturated, typically aromatic, hydrocarbon group that may be a single ring or multiple rings fused together. Examples of aryl groups include, but are not limited to, phenyl, naphthyl.

The term "heteroaryl" refers to a monocyclic or polycyclic aryl group wherein at least one ring atom is a heteroatom selected from N, O, S, and also includes nitrogen oxides thereof when the heteroaryl group contains an N atom. The polycyclic heteroaryl is a fused heteroaryl including, but not limited to, benzo 5-6 membered heteroaryl, 5-6 membered heteroaryl and 5-6 membered heteroaryl. The heteroaryl groups include 5-6 membered heteroaryl, 7-10 membered bicyclic fused heteroaryl, 8-14 membered fused heteroaryl. Examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isoxazolyl, pyridyl, pyrimidinyl, indazolyl, indolyl, isoquinolyl, quinoxalinyl, benzoxazolyl, benzofuranyl, benzothienyl, benzothiazolyl, benzimidazolyl, quinolinyl, quinazolinyl. Examples of 5-6 membered heteroaryl groups include pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isoxazolyl, pyridinyl, pyrimidinyl. Examples of 7-10 membered bicyclic fused heteroaryl groups include indolyl, isoquinolyl, quinoxalinyl, benzoxazolyl, benzofuranyl, benzothienyl, benzothiazolyl, benzimidazolyl, quinolinyl, quinazolinyl.

The term "cycloalkenyl" refers to a group in which at least one ring-forming bond in the cycloalkyl is a double bond and is not aromatic, cycloalkyl being as defined above; cycloalkenyl is preferably 4-, 5-, 6-, 7-, membered cycloalkenyl; examples of 4-membered cycloalkenyl groups include cyclobutenyl, cyclobutenyl; examples of 5-membered cycloalkenyl groups include cyclopentene, cyclopentadiene; examples of 6-membered cycloalkenyl groups include cyclohexenyl, cyclohexadienyl; examples of 7-membered cycloalkenyl groups include cycloheptenyl, cycloheptadienyl. The term "substituted" means that one or more hydrogen atoms on a particular group are replaced with a substituent, provided that the valence of the particular group is normal and the substituted compound is stable.

The term "optionally" means that the substituent may or may not be substituted with other substituents.

The term "composition" as used herein is intended to include products comprising the specified amounts of the respective specified ingredients, as well as any product that results, directly or indirectly, from combination of the specified amounts of the respective specified ingredients. One skilled in the art can vary the actual dosage level of each active ingredient in the pharmaceutical compositions of the present invention so that the resulting amount of active compound is effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration.

The term "pharmaceutically acceptable carrier" refers to a medium commonly accepted in the art for delivering biologically active agents to animals, particularly mammals.

The term "excipient" generally refers to the carrier, diluent, and/or medium required to make an effective pharmaceutical composition.

The term "effective amount" refers to a compound of the invention or a pharmaceutically acceptable salt thereof, which refers to a sufficient amount of the compound to treat a disorder at a reasonable effect/risk ratio applicable to any medical treatment and/or prophylaxis.

The term "pharmaceutically acceptable salt" refers to salts that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like commensurate with a reasonable effect/risk ratio.

In the present invention, the substituents areRepresents the attachment position of the substituent to the parent structure or other fragment. The dash "-" appearing in the substituent structure indicates the point of attachment for the substituent, e.g., -CH ₃, through a C atom. Represents the absolute configuration of one stereocenter, i.e., the R or S configuration.

Where a bond of a substituent may cross-connect to two atoms on a ring, the substituent may be bonded to any atom on the ring. For example, structural unitsIt is meant that the substituent R may be substituted at any position on the phenyl ring.

"Isomers" as used herein include geometric isomers as well as stereoisomers, such as atropisomers, cis-trans isomers, enantiomers, diastereomers, tautomers, and racemic and other mixtures thereof, all of which are within the scope of the present invention. The term "enantiomer" refers to stereoisomers that are mirror images of each other. The term "tautomer" refers to one of the functional group isomers that has a different point of attachment of hydrogen through displacement of one or more double bonds, for example, the ketone and its enol form are keto-enol tautomers. The term "diastereoisomer" refers to a stereoisomer of a molecule having two or more chiral centers and having a non-mirror image relationship between the molecules. The term "cis-trans isomer" refers to different spatial configurations in which either a double bond or a single bond of a ring-forming carbon atom in the molecule cannot rotate freely. The term "atropisomer" refers to stereoisomers that are able to separate out due to the single bond rotation being hindered or rotation being very slow.

Stereoisomers of the compounds of the invention may be prepared by chiral syntheses or chiral reagents or other conventional techniques. For example, one enantiomer of a compound of the invention may be prepared by asymmetric catalytic techniques or chiral auxiliary derivatization techniques. Or by chiral resolution techniques, a single configuration of the compound is obtained from the mixture. Or directly prepared by chiral starting materials. The separation of the optically pure compounds in the invention is usually accomplished by using preparative chromatography, and chiral chromatographic columns are used to achieve the purpose of separating chiral compounds.

In the examples of the present invention, "eluent: tetrahydrofuran/petroleum ether "means that the eluent is composed of a mixture of two solvents, tetrahydrofuran, petroleum ether, and one skilled in the art can determine the appropriate solvent ratio by simple attempts. In the invention, the reagent concentration unit M refers to mol/L;1mM means 1mmol/L.

The chemical abbreviations used in the present invention and their chemical names are as follows:

Detailed Description

The present invention is further illustrated in detail below by means of specific preparation examples and biological experiments, but it should be understood that these examples and biological experiments are for illustrative purposes only and should not be construed as limiting the invention in any way. It will be clear to a person skilled in the art that hereinafter, unless specifically indicated, the materials used are known in the art and may be obtained commercially or by a person skilled in the art according to published or conventional methods. All reactions of the present invention, unless otherwise indicated, were carried out under continuous magnetic stirring under a dry nitrogen or argon atmosphere, the solvent being a dry solvent, wherein: (i) The temperature is expressed in degrees celsius (°c), and the operation is performed at room temperature, which generally refers to 1.35 ℃, preferably 20-30 ℃, more preferably 20-25 ℃; (ii) The solvent is removed by decompression evaporation by a rotary evaporator, and the bath temperature is generally not higher than 60 ℃; (iii) the reaction process is followed by Thin Layer Chromatography (TLC); (iv) The final product had satisfactory hydrogen nuclear magnetic resonance (¹ H-NMR) and/or Mass Spectrometry (MS) data.

Test instrument:

The structure of the compounds of the present invention is determined by Nuclear Magnetic Resonance (NMR) or/and liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using Bruker Neo 400M or Bruker Ascend 400 nuclear magnetic instruments with deuterated dimethyl sulfoxide (DMSO-d 6), deuterated methanol (CD 3 OD) and/or deuterated chloroform (CDCl 3) as the measurement solvent and Tetramethylsilane (TMS) as the internal standard.

The LC-MS was measured by using a Shimadzu 2030Plus-LCMS2020 mass spectrometer, an Agilent 1260-6125B single quadrupole mass spectrometer or a Shimadzu LCMS-2020 mass spectrometer. HPLC was performed using Shimadzu LCMS-2020 or Agilent 1260 high performance liquid chromatography.

Preparation of high performance liquid chromatography using Shimadzu FRC-40equipped with LC-20AP and PDA-20A (column: synergy Max-RP, 150X 30mm,4 m) or GILSON Trilution LC (column: sunFire Prep C18,10 μm, 19X 250mm XBridge Prep C18,10 μm, 19X 250 mm)).

The model number of the microwave synthesizer is Biotage Initiator +.

Preparation example 1: preparation of intermediate A-2:

Step1: synthesis of Compound A-2-2

A-2-1 (45.0 g,265 mmol) and cesium carbonate (172 g,530 mmol) were dissolved in acetonitrile (500 mL), methyl iodide (45.0 g,318 mmol) was added dropwise at 25℃and then reacted at 25℃for 16 hours. The solid was removed by filtration, the filtrate was concentrated under reduced pressure to remove the solvent, and the residue was poured into water (1.5L) and then extracted with ethyl acetate (2L). The organic phase was washed with saturated brine (1L), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give A-2-2.

Step 2: synthesis of Compound A-2-3:

A-2-2 (36 g,196 mmol), ammonium acetate (3.02 g,39.2 mmol) and acetic acid (4.70 g,78.4 mmol) were dissolved in toluene (500 mL), and malononitrile (19.40 g, 254 mmol) was added dropwise and the reaction solution was reacted at 110℃for 48 hours. The reaction mixture was quenched with water (800 mL), extracted with ethyl acetate (1L), and the organic phase was washed with saturated brine (1L), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give the compound A-2-3.¹H NMR(400MHz,DMSO-d₆)δ:4.10(dd,J＝7.04,2.86Hz,2H),2.57(td,J＝3.69,1.87Hz,2H),2.40-2.46(m,2H),1.98-1.90(m,1H),1.78-1.70(m,2H),1.70-1.61(m,1H),1.43(s,3H),1.18(t,J＝7.04Hz,3H).

Step 3.1: synthesis of Compound A-2-4:

Compound A-2-3 (75 g,323 mmol), sublimed sulfur powder (20.70 g, 640 mmol) and L-proline (7.43 g,64.6 mmol) were added to N, N-dimethylformamide (500 mL) at 25℃and then stirred at 80℃for 16 hours. The solvent was removed under reduced pressure, and the residue was poured into water (800 mL) and then extracted with ethyl acetate (1L). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by recrystallization from methylene chloride (200 mL) to give the compound A-2-4.MS m/z(ESI):264.9[M+H]⁺.¹H NMR(400MHz,CDCl₃)δ:4.62(br s,2H),4.32-4.12(m,2H),2.64-2.47(m,2H),2.21(ddd,J＝13.15,7.32,3.96Hz,1H),1.93-1.79(m,2H),1.74-1.66(m,1H),1.61(s,3H),1.31(t,J＝7.15Hz,3H).

Step 3.2: synthesis of Compounds A-2-4-P1, A-2-4-P2:

Compound A-2-4 (6 g,22.7 mmol) was separated by preparative high performance liquid chromatography (column: DAICEL CHIRALPAK AD (250 mm. Times.50 mm,10 μm), mobile phase carbon dioxide-ethanol (0.1% ammonia; gradient: 20% ethanol, flow rate: 200 mL/min) to give compound A-2-4-P1 (peak 1, retention time 0.924 min), compound A-2-4-P2 (peak 2, retention time 1.014 min).

Step 4: synthesis of Compound A-2:

To a mixture of A-2-4 (16 g,60.6 mmol) in ethanol (30 mL) and tetrahydrofuran (30 mL) was added dropwise an aqueous solution (20 mL) of potassium hydroxide (30.50 g,545 mmol) at 25℃and then stirred at 60℃for 16 hours. The reaction solution was cooled to 25 ℃, diluted with water (100 ml), cooled to 0 ℃, and adjusted to pH 5 with 2M hydrochloric acid solution. Then extracted with ethyl acetate (50 mL). The organic phase was washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was purified by slurrying with acetonitrile (100 mL) at 25℃and filtered to give the compound A-2.MS m/z(ESI):237.0[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:12.45(br s,1H),6.87(s,2H),2.42(t,J＝6.00Hz,2H),1.91-1.99(m,1H),1.79-1.69(m,2H),1.68-1.58(m,1H),1.42(s,3H).

Step 5: preparation of Compound A-2-P1

The same method as in step 4 of this example was used to prepare compound A-2-P1 from compound A-2-4-P1.

Example 1 preparation of Compound 1, compound 1a, compound 1b

Step 1: synthesis of Compounds 1-3

A solution of Compound 1-1 (8.40 g,40.38 mmol), compound 1-2 (5.75 mL,40.38 mmol), cuprous iodide (0.14 mL,4.04 mmol), bis-triphenylphosphine palladium dichloride (1.42 g,2.02 mmol) and triethylamine (16.79 mL,121.15 mmol) in acetonitrile (100 mL) was stirred under nitrogen at 80℃for 12 hours. The reaction solution was filtered, diluted with water (100 mL), and extracted with ethyl acetate (150 mL. Times.3). The organic phase was washed with saturated brine (100 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by silica gel column chromatography to obtain the compound 1-3.MS m/z (ESI): 179.5[ M+H ] ⁺.

Step 2: synthesis of Compounds 1-4

To a solution of compounds 1-3 (5.60 g,31.41 mmol) in ethanol (100 mL) was added potassium carbonate (13.02 g,94.22 mmol) followed by stirring at 30℃for 12 hours. Concentrating the reaction solution, and purifying the crude product by silica gel column chromatography to obtain the compounds 1-4.MS m/z (ESI): 107.2[ M+H ] ⁺.

Step 3: synthesis of Compounds 1-6

To an acetic acid solution (300 mL) of compounds 1 to 5 (30 g,184.05 mmol) and potassium acetate (18.06 g,184.05 mmol) was added iodine monochloride (18.68 mL,368.10 mmol), and the reaction solution was stirred at 30℃for 16 hours. Diluting with water (500 mL), filtering, drying and concentrating to obtain compounds 1-6. ¹H NMR(400MHz,DMSO-d₆ ) Delta 6.87 (br s, 2H), 6.57 (s, 1H).

Step 4: synthesis of Compounds 1-7

A solution of compounds 1-6 (6.81 g,23.56 mmol), compounds 1-4 (2.50 g,23.56 mmol), cuprous iodide (0.08 mL,2.36 mmol), bis triphenylphosphine palladium dichloride (0.83 g,1.18 mmol) and triethylamine (6.53 mL,47.12 mmol) in acetonitrile (100 mL) was stirred under nitrogen at 80℃for 12 hours. The reaction solution is filtered and concentrated, and the crude product is purified by silica gel column chromatography to obtain the compounds 1-7.MS m/z (ESI): 268.8[ M+H ] ⁺.

Step 5: synthesis of Compounds 1-8

To a solution of compounds 1-7 (3.95 g,14.79 mmol) in N, N-dimethylformamide (40 mL) was added potassium tert-butoxide (3.65 mL,29.58 mmol), and the reaction was stirred at 100deg.C for 2 hours. The reaction solution was filtered, diluted with water (50 mL), and extracted with ethyl acetate (50 mL. Times.3). The organic phase was washed with saturated brine (50 mL. Times.2), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated to give the compound 1-8.¹H NMR(400MHz,DMSO-d₆)δ:12.42(s,1H),7.95(s,1H),7.82(d,J＝4.0Hz,1H),7.36(s,1H),6.86(d,J＝4.0Hz,1H),3.94(s,3H).

Step 6: synthesis of Compounds 1-9

To a solution of compounds 1 to 8 (1.20 g,4.49 mmol) in tetrahydrofuran (15 mL) was added sodium hydride (0.27 g,6.74 mmol), followed by addition of methyl iodide (0.28 mL,4.49 mmol) to the reaction solution. The final reaction solution was stirred at 50℃for 1 hour. The reaction solution is concentrated, and the crude product is purified by silica gel column chromatography to obtain the compounds 1-9.MS m/z (ESI): 282.8[ M+H ] ⁺.

Step 7: synthesis of Compounds 1-11

To a solution of compound A-1 (399mg, 3.06 mmol) in N, N-dimethylformamide (10 mL) was added sodium hydride (183.54 mg,4.59 mmol), and after 30 minutes of reaction, compound 1-9 (860.0 mg,3.06 mmol) was added. The final reaction solution was stirred at 80℃for 48 hours. The reaction mixture was quenched with saturated ammonium chloride solution (10 mL) and extracted with ethyl acetate (15 mL. Times.3). The organic phase was washed with saturated brine (10 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, purifying the crude product by silica gel column chromatography to obtain the compound 1-11.MS m/z(ESI):375.2[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.40(s,1H),6.90(s,1H),6.74(s,1H),6.52(s,1H),5.56-5.47(m,1H),3.96(s,3H),3.95(s,3H),2.63-2.54(m,3H),2.37-2.26(m,1H),1.91-1.55(m,6H),1.41(s,3H).

Step 8: synthesis of Compounds 1-12

A solution of compounds 1-11 (750.0 mg,2.01 mmol), zinc cyanide (1884 mg,16.05 mmol) and bis (tri-tert-butylphosphine) palladium (102.52 mg,0.20 mmol) in N-methylpyrrolidone (10 mL) was stirred under nitrogen at 120℃for 20 min under microwaves. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (20 mL. Times.3). The organic phase was washed with saturated brine (10 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by silica gel column chromatography to obtain the compounds 1-12.MS m/z (ESI): 365.2[ M+H ] ⁺.

Step 9: synthesis of Compounds 1-13

To an ethanol solution (10 mL) of compounds 1-12 (600.0 mg,1.65 mmol) and hydroxylamine hydrochloride (297.5 mg,4.28 mmol) was added sodium carbonate (226.8 mg,2.14 mmol), and the final reaction was stirred at 80℃for 1 hour. The reaction solution is directly concentrated to obtain the compounds 1-13.MS m/z (ESI): 398.3[ M+H ] ⁺.

Step 10: synthesis of Compounds 1-14

To a solution of compound A-2 (166.5 mg,0.70 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (267.86 mg,0.70 mmol) in N, N-dimethylformamide (5 mL) was added N, N-diisopropylethylamine (0.35 mL,2.11 mmol), and after stirring for 30 minutes, compound 1-13 (280.0 mg,0.70 mmol) was added. The final reaction solution was stirred at 20℃for 11.5 hours. The reaction solution is concentrated, and the crude product is purified by silica gel column chromatography to obtain the compounds 1-14.MS m/z (ESI): 616.5[ M+H ] ⁺.

Step 11: synthesis of Compound 1

A solution of compounds 1-14 (300.0 mg,0.49 mmol) and sodium carbonate (154.9 mg,1.46 mmol) in N, N-dimethylformamide (5 mL) was stirred at 100deg.C for 12 hours. The reaction solution was concentrated to obtain a crude product. Purification by preparative high performance liquid chromatography (column: xtimate C18,150 X40 mm,5 μm; mobile phase: 0.225% aqueous formic acid-acetonitrile; gradient: 25% -95% acetonitrile, 18 min) gives compound 1.MS m/z (ESI): 598.1[ M+H ] ⁺.

Step 12: separation of Compound 1a and Compound 1b

Compound 1 (35.4 mg,0.06 mmol) was separated by preparative high performance liquid chromatography (instrument: gilson-281, column: IG 20X 250mm,10 μm (Daicel), mobile phase: n-hexane (0.1% diethylamine)/ethanol (0.1% diethylamine) =65/35, flow rate: 45 mL/min) to give compound 1a (peak 1, retention time 7.58 min) and compound 1b (peak 2, retention time 10.05 min).

Compounds of formula (I) 1a：MS m/z(ESI):598.4[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.83(d,J＝4.0Hz,2H),7.09(s,2H),6.85(s,1H),6.83(d,J＝4.0Hz,1H),5.55-5.51(m,1H),4.13(s,3H),3.94(s,3H),3.05-3.02(m,1H),2.58-2.55(m,2H),2.49(s,3H),2.18-2.11(m,2H),1.91-1.71(m,11H),1.35(d,J＝8.0Hz,3H).

Compounds of formula (I) 1b：MS m/z(ESI):598.4[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.83(d,J＝4.0Hz,2H),7.08(s,2H),6.85(s,1H),6.83(d,J＝4.0Hz,1H),5.55-5.52(m,1H),4.13(s,3H),3.94(s,3H),3.01-2.97(m,1H),2.58-2.55(m,2H),2.49(s,3H),2.19-2.07(m,2H),1.96-1.68(m,11H),1.33(d,J＝4.0Hz,3H).

Example 2: preparation of Compound 2

Step 1: synthesis of Compound 2-2:

Compound 2-1 (1.40 g,5.71 mmol) was dissolved in phosphorus oxychloride (56 mL), N-diisopropylethylamine (1.11 g,8.56 mmol) was added dropwise at 25℃and then reacted in an oil bath at 110℃for 3 hours. Removing solvent under reduced pressure, pouring the residual oil into ice water, adding saturated sodium bicarbonate solution to adjust pH to weak alkalinity, extracting with dichloromethane (50 mL), drying with anhydrous sodium sulfate, filtering, and removing solvent under reduced pressure to obtain compound 2-2.MS m/z(ESI):282.0[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.30(d,J＝8.4Hz,1H),7.99(s,1H),7.94(d,J＝8.8Hz,1H),3.77-3.49(m,2H),3.18-3.07(m,1H),2.21(s,3H),1.99-1.83(m,2H),1.31-1.23(m,2H).

Step 2: synthesis of Compound 2-3:

compound 2-2 (1.57 g,5.59 mmol) was dissolved in N, N-dimethylformamide (30 mL), and (S) - (+) -3-hydroxy piperidine hydrochloride (827.0 mg,6.01 mmol) and N, N-diisopropylethylamine (1.62 g,12.5 mmol) were added and reacted at 25℃for 16 hours. Water (50 mL) was added to the reaction mixture, the mixture was extracted with ethyl acetate (80 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a flash silica gel column to give the compound 2-3.MS m/z(ESI):347.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.02(d,J＝8.40Hz,1H),7.58(s,1H),7.50(dd,J＝8.80,1.20Hz,1H),5.08-4.96(m,1H),4.10(d,J＝12.40Hz,1H),3.95-3.84(m,1H),3.73(dd,J＝6.80,3.60Hz,1H),3.63-3.49(m,1H),3.38(td,J＝8.40,4.00Hz,1H),3.29-3.15(m,2H),2.35-2.16(m,2H),2.12(d,J＝1.80Hz,3H),1.98-1.74(m,4H),1.67-1.50(m,3H).

Step 3: synthesis of Compounds 2-4:

Compound 2-3 (400.0 mg,1.15 mmol) was dissolved in dimethyl sulfoxide (6 mL), triethylenediamine (129.0 mg,1.15 mmol) was added and stirred at 25℃for 1 hour, and sodium cyanide (80 mg,1.63 mmol) was added and reacted at 80℃for 2 hours. Water (20 mL) was added, followed by extraction with ethyl acetate (20 mL. Times.3), the aqueous phase was extracted 3 times with ethyl acetate (10 mL), the combined organic phases were washed with saturated brine (50 mL), the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the compound 2-4.MS m/z(ESI):338.0[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.09(d,J＝8.80Hz,1H),7.75(s,1H),7.65(dd,J＝8.40,1.20Hz,1H),5.03(t,J＝3.60Hz,1H),4.12(d,J＝12.00Hz,1H),3.90(dd,J＝10.40,1.60Hz,1H),3.80-3.70(m,1H),3.67-3.56(m,1H),3.49-3.39(m,2H),3.25-3.17(m,1H),2.34-2.21(m,2H),2.14(d,J＝2.40Hz,3H),1.95-1.78(m,4H),1.66-1.52(m,3H).

Step 4: synthesis of Compounds 2-5:

Compound 2-4 (150.0 mg,0.44 mmol) was dissolved in absolute ethanol (3 mL), hydroxylamine hydrochloride (80.3 mg,1.16 mmol) and sodium carbonate (61.3 mg,0.58 mmol) were added and reacted at 85℃for 2 hours. Water (10 mL) was added, extraction was performed using ethyl acetate (10 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 2-5.

Step 5: synthesis of Compounds 2-6:

Compound 2-5 (164.0 mg, 443. Mu. Mol) and compound A-2 (87.0 mg, 368. Mu. Mol) were dissolved in N, N-dimethylformamide (2 mL), N-diisopropylethylamine (143 mg,1.11 mmol) was added dropwise, and after cooling to 0℃with ice and stirring for 0.5 hours, benzotriazol-1-yl-oxy-tripyrrolidinylphosphine hexafluorophosphate (288 mg, 553.4. Mu. Mol) was added, followed by stirring to 25℃for 5.5 hours. Water (10 mL) was added to the reaction solution, extraction was performed using ethyl acetate (10 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified using a flash silica gel column to obtain compounds 2 to 6.MS m/z (ESI): 589.3[ M+H ] ⁺.

Step 6: synthesis of Compound 2:

Compounds 2-6 (60 mg, 101. Mu. Mol) were dissolved in N, N-dimethylformamide (2 mL), and anhydrous sodium carbonate (21.6 mg, 204. Mu. Mol) was added thereto and reacted at 100℃for 16 hours. Purifying with preparative high performance liquid chromatograph after filtering to obtain compound 2.MS m/z(ESI):571.3[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.06(d,J＝7.60Hz,1H),7.85(s,1H),7.60(d,J＝7.60Hz,1H),7.10(s,2H),5.02(s,1H),4.23-4.09(m,1H),4.07-3.91(m,1H),3.77(d,J＝2.40Hz,1H),3.56-3.45(m,2H),3.26-3.12(m,2H),2.62-2.55(m,2H),2.38-2.21(m,2H),2.15(s,3H),2.09(d,J＝8.80Hz,1H),2.02-1.76(m,10H),1.67(s,2H),1.59-1.48(m,1H).

Example 3: preparation of Compound 3

Step 1: synthesis of Compound 3-2

To a solution of compound A-3 (2.0 g,10.69 mmol) in N, N-dimethylformamide (20 mL) was slowly added sodium hydride (641.6 mg,16.04 mmol) in portions under nitrogen at 0deg.C. After the addition was completed, the mixture was stirred at 25℃for half an hour. Methyl iodide (1.33 mL,21.39 mmol) was finally added and the reaction stirred at 25℃for two hours. The reaction mixture was slowly quenched with saturated ammonium chloride solution (5 mL) and diluted with water (150 mL), extracted with ethyl acetate (50 mL. Times.3), and the combined organic phases were washed with water (200 mL) and saturated brine (300 mL), respectively. Drying with anhydrous sodium sulfate, and concentrating under reduced pressure 3-2.MS m/z(ESI):201.0[M+H]⁺.¹H NMR(400MHz,Methanol-d₄)δ:7.51(d,J＝1.20Hz,1H),7.40(d,J＝4.0Hz,1H),6.61(dd,J＝4.0,4.0Hz,1H),3.85(s,3H).

Step 2: synthesis of Compound 3-3

To a solution of compound A-1 (578.4 mg,4.48 mmol) in tetrahydrofuran (10 mL) was added slowly in portions sodium hydride (268.6 mg,6.71 mmol) under nitrogen at 25 ℃. After the addition was completed, the mixture was stirred at 25℃for 10 minutes. Finally, compound 3-2 (900.0 mg,4.48 mmol) was added, and the mixture was heated to 80℃and stirred for 18 hours. The reaction mixture was slowly quenched with saturated ammonium chloride solution (5 mL) and diluted with water (300 mL), extracted with ethyl acetate (50 mL. Times.2), and the combined organic phases were washed with water (200 mL) and saturated brine (200 mL), respectively. Drying anhydrous sodium sulfate, concentrating under reduced pressure to obtain residue, and purifying by flash silica gel column chromatography to obtain compound 3-3.MS m/z(ESI):294.2[M+H]⁺.¹H NMR(400MHz,Methanol-d₄)δ:7.12(d,J＝1.2Hz,1H),7.07(s,1H),6.52(d,J＝4.0Hz,1H),5.38(p,J＝8.0Hz,1H),3.77(s,3H),3.08-3.03(m,1H),2.73–2.69(m,1H),2.55(s,3H),2.39–2.33(m,1H),2.09–1.98(m,1H),1.85–1.72(m,3H),1.39(d,J＝4.0Hz,3H).

Step 3: synthesis of Compounds 3-4

In a microwave reactor, compound 3-3 (960.0 mg,3.27 mmol), bis (tri-t-butylphosphine) palladium (670.6 mg,1.31 mmol) and zinc cyanide (1918.4 mg,16.34 mmol) were dissolved in N-methylpyrrolidone (20 mL) and heated to 160℃under nitrogen atmosphere and reacted for 90 minutes with stirring. The reaction solution is filtered by diatomite, the mother solution is decompressed and concentrated to obtain a residue, and the residue is purified by flash silica gel column chromatography to obtain 3-4.MS m/z (ESI): 285.2[ M+H ] ⁺.

Step 4: synthesis of Compound 3-5

A mixture of Compound 3-4 (290.0 mg,1.02 mmol), hydroxylamine hydrochloride (70.9 mg,1.02 mmol), sodium carbonate (118.9 mg,1.12 mmol) and ethanol (3 mL) was heated to 80℃and reacted with stirring for 2 hours. The reaction solution was diluted with ethanol (2 mL), filtered and concentrated under reduced pressure to give 3-5.MS m/z (ESI): 318.2[ M+H ] ⁺.

Step 5: synthesis of Compounds 3-6

A solution of compound 3-5 (320.0 mg,1.01 mmol), compound A-2 (238.2 mg,1.01 mmol), 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (402.5 mg,1.06 mmol) and N-ethyldiisopropylamine (0.5 mL,3.02 mmol) in N, N-dimethylformamide (5 mL) was stirred at 25℃for 18 hours. The reaction solution is filtered and then is directly purified by a C18 reversed phase chromatographic column to obtain 3-6.MS m/z (ESI): 536.4[ M+H ] ⁺.

Step 6: synthesis of example 3

A solution of compound 3-6 (140.0 mg,0.26 mmol) and sodium carbonate (166.2 mg,1.57 mmol) in N, N-dimethylformamide (6 mL) was heated to 120℃and reacted with stirring for 18 hours. The reaction solution was filtered and purified by high performance liquid chromatography (column: synergy Max-RP, 150X 30mm,4 μm; mobile phase: 0.075% aqueous trifluoroacetic acid/acetonitrile=65/35, flow rate: 30 mL/min) to give Compound 3 (trifluoroacetate salt) ).MS m/z(ESI):518.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:10.14(s,1H),7.95(d,J＝8.0Hz,1H),7.55(d,J＝4.0Hz,1H),7.10(s,2H),6.66(d,J＝4.0Hz,1H),5.48–5.38(m,1H),3.93(d,J＝8.0Hz,3H),3.85–3.78(m,1H),3.65–3.57(m,1H),3.19–3.11(m,1H),3.05–2.98(m,3H),2.59–2.55(m,2H),2.34–2.26(m,1H),2.15–2.06(m,2H),2.01–1.86(m,5H),1.83(s,3H),1.54–1.47(m,3H).

Example 4: preparation of Compound 4

Step 1: synthesis of Compound 4-2

Triethylamine (4.67 mL,33.66 mmol) was added to a mixed solution of compound 4-1 (3 g,16.83 mmol) and 4-methylbenzenesulfonyl chloride (3.21 mL,16.83 mmol) in methylene chloride (30 mL), and stirred at 30℃for 12 hours. Concentrating the reaction solution, and purifying the crude product by silica gel column chromatography to obtain the compound 4-2.MS m/z (ESI): 355.2[ M+Na ] ⁺.

Step 2: synthesis of Compound 4-3

Cesium carbonate (4.18 g,12.83 mmol) was added to a mixed solution of compound A-3 (1.2 g,6.42 mmol) and compound 4-2 (2.13 g,6.42 mmol) in N, N-dimethylformamide (20 mL) and stirred at 100deg.C for 12 hours. The reaction mixture was diluted with water (20 mL), and extracted with ethyl acetate (20 mL. Times.3). The organic phase was washed with saturated brine (20 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by silica gel column chromatography to obtain the compound 4-3.MS m/z (ESI): 347.2[ M+H ] ⁺.

Step 3: synthesis of Compound 4-4

To a solution (20 mL) of compound A-1 (539.5 mg,4.18 mmol) in N, N-dimethylformamide was added sodium hydride (250.6 mg,6.26 mmol), and after 30 minutes the reaction was continued, compound 4-3 (1450.0 mg,4.18 mmol) was added. The final reaction solution was stirred at 80℃for 15.5 hours. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL. Times.3). The organic phase was washed with saturated brine (20 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by silica gel column chromatography to obtain the compound 4-4.MS m/z (ESI) 440.4[ M+H ] ⁺.

Step 4: synthesis of Compound 4-5

A solution of compound 4-4 (1 g,2.27 mmol), zinc cyanide (2.14 g,18.18 mmol), and di-tri-tert-butylphosphine palladium (0.12 g,0.23 mmol) in N-methylpyrrolidone (15 mL) was stirred under nitrogen at 120℃for 20 minutes in a microwave atmosphere. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (25 mL. Times.3). The organic phase was washed with saturated brine (20 mL. Times.2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give the crude product which was purified by silica gel column chromatography to give Compound 4-5.MS m/z (ESI): 431.2[ M+H ] ⁺.

Step 5: synthesis of Compounds 4-6

To a solution of compound 4-5 (800 mg,1.86 mmol) and hydroxylamine hydrochloride (335.71 mg,4.83 mmol) in ethanol (10 mL) was added sodium carbonate (256.02 mg,2.42 mmol), and the reaction was stirred at 80℃for 1 hour. The reaction solution is concentrated, and the crude product is purified by silica gel column chromatography to obtain the compound 4-6.MS m/z (ESI): 464.4[ M+H ] ⁺.

Step 6: synthesis of Compounds 4-7

To a solution of compound A-2 (254.9 mg,1.08 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (410.1 mg,1.08 mmol) in N, N-dimethylformamide (10 mL) was added N, N-diisopropylethylamine (0.54 mL,3.24 mmol), and after stirring for 30 minutes, compound 4-6 (500.0 mg,1.08 mmol) was added. The reaction solution was stirred at 20℃for 11.5 hours. The reaction solution is concentrated, and the crude product is purified by silica gel column chromatography to obtain the compound 4-7.MS m/z (ESI): 682.9[ M+H ] ⁺.

Step 7: synthesis of Compounds 4-8

To a solution of compound 4-7 (300.0 mg,0.44 mmol) in N, N-dimethylformamide (6 mL) was added sodium carbonate (139.9 mg,1.32 mmol), and the resulting reaction mixture was stirred at 100℃for 12 hours. Concentrating the reaction solution, and purifying by silica gel column chromatography to obtain compounds 4-8.MS m/z (ESI): 664.4[ M+H ] ⁺.

Step 8: synthesis of Compound 4

To a solution of compounds 4-8 (100.0 mg,0.15 mmol) in methylene chloride (2 mL) was added trimethyliodosilane (0.06 mL,0.45 mmol), and the reaction was stirred at 20℃for 20 min. The reaction solution was concentrated to obtain a crude product. Purifying with high performance liquid chromatography (chromatographic column: synergi Max-RP, 150X30 mm,4 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 20% -50%,19 min) to obtain compound 4.MS m/z(ESI):574.0[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.14(s,1H),7.80(d,J＝4.0Hz,1H),7.76(d,J＝4.0Hz,1H),7.08(s,2H),6.64(s,1H),5.46(t,J＝4.0Hz,1H),5.37(d,J＝4.0Hz,1H),5.28(t,J＝4.0Hz,1H),4.48-4.42(m,2H),2.68-2.61(m,5H),2.33-2.32(m,2H),2.11-1.89(m,6H),1.86-1.74(m,8H),1.37(d,J＝4.0Hz,3H).

Example 6: preparation of Compound 6

Step 1: synthesis of Compound 6-1

Compound A-3 (1.5 g,8.02 mmol) was dissolved in tetrahydrofuran (20 mL), sodium hydride (480 mg,12.03 mmol) was added at 0deg.C, and the mixture was warmed to room temperature and stirred for 30 minutes, and iodocyclopentane (2.36 g,12.03 mmol) was added to the reaction solution. The reaction solution was raised to 80℃under nitrogen atmosphere and stirred for 18 hours. The reaction mixture was quenched by pouring it into 20mL of saturated ammonium chloride, and extracted with ethyl acetate (25 mL. Times.2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give Compound 6-1.MS m/z (ESI): 255.0[ M+H ] ⁺.

Step 2: synthesis of Compound 6-2

Compound A-1 (500 mg,3.86 mmol) was dissolved in tetrahydrofuran (30 mL), cooled to 0 ℃, sodium hydride (155 mg,3.86 mmol) was added, warmed to room temperature, after stirring for 30 minutes, compound 6-1 (820 mg,3.22 mmol) was added to the system, and the mixture was warmed to 80℃and stirred for 16 hours. The reaction mixture was quenched by pouring it into 20mL of saturated ammonium chloride, and extracted with ethyl acetate (25 mL. Times.2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give the compound 6-2.MS m/z(ESI):348.2[M+H]⁺.¹H NMR(400MHz,CDCl₃)δ:7.02(d,J＝3.2Hz,1H),6.91(d,J＝0.8Hz,1H),6.53(dd,J＝3.3,0.9Hz,1H),5.42(p,J＝6.4Hz,1H),4.67–4.57(m,1H),3.09(ddd,J＝9.6,4.4,1.6Hz,1H),2.64(dt,J＝8.4,6.0Hz,1H),2.50(s,3H),2.32–2.12(m,3H),1.96–1.65(m,10H),1.36(d,J＝6.4Hz,3H).

Step 3: synthesis of Compound 6-3

Compound 6-2 (1300 mg,3.73 mmol) and zinc cyanide (4360 mg,37.3 mmol) were dissolved in N-methylpyrrolidone (20 mL) and ditri-butylphosphine palladium (100 mg,0.2 mmol) was added under nitrogen. The reaction solution was heated to 160℃in a microwave and stirred for 1 hour. The reaction mixture was poured into 20mL of water, and extracted with ethyl acetate (55 mL. Times.2). The organic phases were combined, washed 5 times with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give the compound 6-3.MS m/z(ESI):339.2[M+H]⁺.¹H NMR(400MHz,CDCl₃)δ:7.42(d,J＝0.8Hz,1H),7.24(d,J＝3.2Hz,1H),6.64(d,J＝3.2Hz,1H),5.46(t,J＝6.4Hz,1H),4.72(m,1H),3.09(d,J＝2.4Hz,1H),2.70–2.61(m,1H),2.50(s,3H),2.25(tt,J＝9.6,6.0Hz,3H),1.96–1.69(m,10H),1.36(d,J＝6.4Hz,3H).

Step 4: synthesis of Compound 6-4

Compound 6-3 (650 mg,1.91 mmol) was dissolved in ethanol (8 mL), and sodium carbonate (212 mg,2.0 mmol) and hydroxylamine hydrochloride (141 mg,2.0 mmol) were added. The reaction solution was raised to 90℃and stirred for 16 hours. Sodium carbonate was filtered, the filter cake was washed with dichloromethane (2 mL. Times.3) and methanol (2 mL. Times.3), the organic phases were combined and concentrated to give compound 6-4.MS m/z (ESI): 372.2[ M+H ] ⁺.

Step 5: synthesis of Compound 6-5

Compound 6-4 (710 mg,1.91 mmol) was dissolved in N, N-dimethylformamide (15 mL), and Compound A-2 (450 mg,1.91 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (1450 mg,3.82 mmol), N, N-diisopropylethylamine (740 mg,5.73 mmol) were added. Stirring was carried out at room temperature for 18 hours. The reaction mixture was quenched with 20mL of water, and extracted with ethyl acetate (25 mL. Times.2). The organic phases were combined, washed 3 times with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography to give Compound 6-5.MS m/z (ESI): 590.4[ M+H ] ⁺.

Step 6: synthesis of Compound 6

Compound 6-5 (300 mg,0.51 mmol) and sodium carbonate (323 mg,3.05 mmol) were dissolved in N, N-dimethylformamide (10 mL), and the reaction was heated to 120℃and stirred for 16 hours. Adding water into the reaction solution, extracting with ethyl acetate (20 mL×2), mixing the organic phases, drying with anhydrous sodium sulfate, filtering, concentrating, and purifying by high performance liquid chromatography (chromatographic column: synergi Max-RP,150×30mm,4 μm; mobile phase: 0.07% trifluoroacetic acid water solution-acetonitrile; gradient: acetonitrile 35% -65%,19 min), to obtain the compound 6.MS m/z(ESI):572.1[M+H]⁺.¹H NMR(400MHz,Methanol-d₄)δ:8.13(d,J＝1.6Hz,1H),7.65–7.53(m,1H),6.74(d,J＝3.2Hz,1H),5.35–5.17(m,1H),5.04(t,J＝6.8Hz,1H),3.95–3.69(m,2H),3.24–3.03(m,1H),2.85(d,J＝17.2Hz,3H),2.64(t,J＝6.0Hz,2H),2.44(dd,J＝13.2,6.8Hz,1H),2.35–2.25(m,2H),2.25–1.79(m,16H),1.56(dd,J＝6.4,3.2Hz,3H).

Example 7: preparation of Compound 7

Step 1: synthesis of Compound 7-2

Compound 7-1 (7.30 g,38.8 mmol) was dissolved in N, N-dimethylformamide (70 mL), sodium hydride (2.02 g,50.5mmol, purity 60%) was added at 0℃under nitrogen, followed by stirring for 1 hour, 3- (chloromethyl) -1-methyl-1H-pyrazole (6.59 g,50.5 mmol) was added, and the mixture was allowed to react at 25℃for 18 hours. Cooling to 0deg.C, adding dropwise into saturated ammonium chloride solution (100 mL), diluting with water (50 mL), extracting with ethyl acetate (80 mL), drying the organic phase with anhydrous sodium sulfate, filtering, concentrating, and purifying with flash silica gel column to obtain compound 7-2.MS m/z(ESI):281.8[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.78(d,J＝3.52Hz,1H),7.62(d,J＝1.76Hz,1H),6.71(d,J＝3.74Hz,1H),6.17(d,J＝1.76Hz,1H),5.38(s,2H),3.78(s,3H).

Step 2: synthesis of Compound 7-3

Compound 7-2 (3.50 g,12.4 mmol) was dissolved in methanol (40 mL), cooled to 0deg.C, and a 30% sodium methoxide in methanol (2.3 mL,12.4 mmol) was added and reacted at 25deg.C for 3 hours. Diluting with water (50 mL), extracting with ethyl acetate (80 mL), drying the organic phase with anhydrous sodium sulfate, filtering, concentrating, and purifying with flash silica gel column to obtain the compound 7-3.MS m/z(ESI):278.2[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.60(d,J＝2.20Hz,1H),7.44(d,J＝3.52Hz,1H),6.55(d,J＝3.52Hz,1H),6.11(d,J＝2.20Hz,1H),5.31(s,2H),4.05(s,3H),3.78(s,3H).

Step 3: synthesis of Compound 7-4

A-1 (0.98 g,7.56 mmol) and Compound 7-3 (2 g,7.20 mmol) were dissolved in anhydrous tetrahydrofuran (20 mL), cooled to 0deg.C, and sodium t-butoxide (1.38 g,14.4 mmol) was added thereto, and the reaction mixture was reacted at 25deg.C for 18 hours. Diluting with water (50 mL), extracting with ethyl acetate (80 mL), drying the organic phase with anhydrous sodium sulfate, filtering, concentrating, and purifying with flash silica gel column to obtain the compound 7-4.MS m/z(ESI):371.2[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.59(d,J＝2.20Hz,1H),7.16(d,J＝3.52Hz,1H),6.38(d,J＝3.52Hz,1H),6.07(d,J＝2.20Hz,1H),5.31-5.24(m,1H),5.22(s,2H),3.99(s,3H),3.78(s,3H),3.13-2.86(m,1H),2.81-2.57(m,1H),2.48-2.33(m,3H),2.31-2.09(m,1H),1.84-1.63(m,4H),1.28(d,J＝5.72Hz,3H).

Step 5: synthesis of Compound 7-5

Compound 7-4 was dissolved in (750 mg,2.02 mmol) and acetonitrile (10 mL), and after addition of trimethyliodosilane (4816 mg,2.43 mmol), it was reacted at 25℃for 1 hour, followed by heating to 80℃for 3 hours. The reaction solution is cooled to 25 ℃, concentrated under reduced pressure and purified by a rapid silica gel column to obtain the compound 7-5.MS m/z(ESI):357.0[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:11.93(s,1H),7.61(d,J＝1.98Hz,1H),6.94(d,J＝3.30Hz,1H),6.36(d,J＝3.30Hz,1H),6.12(d,J＝2.20Hz,1H),5.16(s,2H),5.10(dd,J＝10.01,6.05Hz,1H),3.78(s,3H),3.69-3.58(m,2H),3.16-3.09(m,1H),3.02(d,J＝5.06Hz,3H),2.29-2.21(m,1H),2.08(dt,J＝13.04,6.79Hz,1H),1.99-1.75(m,2H),1.45(d,J＝6.16Hz,3H).

Step 6: synthesis of Compound 7-6

Compound 7-5 (845 mg,2.37 mmol) was dissolved in toluene (15 mL), phosphorus oxychloride (1.82 g,11.9 mmol) and N, N-diisopropylethylamine (313 mg,4.74 mmol) were added, and then reacted at 80℃for 3 hours. Removing solvent under reduced pressure, slowly adding the residual oil into 50mL of ice water, adjusting pH to about 8-9 with saturated sodium bicarbonate solution, extracting with dichloromethane (20 mL), drying the organic phase with anhydrous sodium sulfate, filtering, and removing solvent under reduced pressure to obtain the compound 7-6.MS m/z(ESI):374.9[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.63(d,J＝1.96Hz,1H),7.56(d,J＝3.67Hz,1H),6.58(d,J＝3.55Hz,1H),6.17(d,J＝2.08Hz,1H),5.32(s,2H),5.26(dd,J＝9.17,6.11Hz,1H),3.79(s,3H),3.60-3.53(m,1H),3.23–3.13(m,2H),3.05-2.98(m,3H),2.28(dd,J＝12.78,5.07Hz,1H),2.13–2.05(m,1H),2.00-1.80(m,2H),1.44(d,J＝6.11Hz,3H).

Step 7: synthesis of Compound 7-7

Compounds 7-6 (600 mg,1.60 mmol) were dissolved in dimethyl sulfoxide (6 mL), triethylenediamine (180 mg,1.60 mmol) was added and stirred at 25℃for 1 hour, and sodium cyanide (150 mg,3.06 mmol) was added and reacted at 80℃for 2 hours. Water (20 mL) was added, the mixture was extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give Compound 7-7.MS m/z (ESI): 366.2[ M+H ] ⁺.

Step 8: synthesis of Compounds 7-8

Compound 7-7 (200 mg,0.55 mmol) was dissolved in absolute ethanol (3 mL), hydroxylamine hydrochloride (99 mg,1.42 mmol) and sodium carbonate (75.4 mg,0.71 mmol) were added and reacted at 85℃for 2 hours. Water (10 mL) was added for dilution, ethyl acetate (10 mL) was used for extraction, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 7-8.

Step 9: synthesis of Compounds 7-9

Compound 7-8 (200 mg,0.50 mmol) and compound A-2 (130 mg,0.55 mmol) were dissolved in N, N-dimethylformamide (3 mL), N-diisopropylethylamine (195 mg,1.51 mmol) was added dropwise, then cooled to 0℃and stirred for 0.5 hours, benzotriazol-1-yl-oxy-tripyrrolidinylphosphine hexafluorophosphate (390 mg,0.75 mmol) was added, and then stirred for 5.5 hours at 25 ℃. Water (10 mL) was added for dilution, ethyl acetate (10 mL) was used for extraction, and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give compounds 7-9.MS m/z (ESI): 617.1[ M+H ] ⁺.

Step 10: synthesis of Compound 7

Compounds 7-9 (308 mg,0.50 mmol) were dissolved in N, N-dimethylformamide (4 mL), and anhydrous sodium carbonate (155 mg,1.46 mmol) was added and reacted at 100℃for 16 hours. Filtering the reaction solution, purifying by preparative high performance liquid chromatography (chromatographic column: boston Prime C18,150 ×30mm,5 μm; mobile phase: 0.05% ammonia water solution (wt 0.08% ammonium bicarbonate) -acetonitrile; gradient: 55% -75%,10 min), to obtain the compound 7.MS m/z(ESI):599.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.61(d,J＝2.20Hz,1H),7.57(d,J＝3.50Hz,1H),7.11(s,2H),6.87(d,J＝3.70Hz,1H),6.13(d,J＝2.00Hz,1H),5.44-5.21(m,3H),3.78(s,3H),3.08-2.95(m,2H),2.78-2.69(m,1H),2.58-2.54(m,2H),2.43(s,3H),2.26-2.10(m,2H),2.02-1.93(m,1H),1.84(s,3H),1.82-1.77(m,2H),1.76-1.65(m,3H),1.30(d,J＝6.40Hz,3H).

Example 9: preparation of Compound 9

Step 1: synthesis of Compound 9-2

Compound 9-1 (15.0 g,79.8 mmol) was dissolved in N, N-dimethylformamide (150 mL), sodium hydride (3.51 g,87.8mmol, purity 60%) was added at 0℃under nitrogen atmosphere, followed by stirring for 1 hour, methyl iodide (12.5 g,87.8 mmol) was added, and the temperature was raised to 25℃for reaction for 18 hours. Cooling to 0deg.C, adding dropwise saturated ammonium chloride solution (300 mL), diluting with water (300 mL), extracting with ethyl acetate (300 mL), drying and concentrating the organic phase with anhydrous sodium sulfate, and purifying with rapid silica gel column to obtain compound 9-2.MS m/z(ESI):202.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.75(d,J＝3.52Hz,1H),6.69(d,J＝3.52Hz,1H),3.84–3.76(m,3H).

Step 2: synthesis of Compound 9-3

Compound 9-2 (6.00 g,29.7 mmol) was dissolved in methanol (50 mL), and a 30% sodium methoxide methanol solution (8.3 mL,44.6 mmol) was added dropwise at 0℃and reacted at 25℃for 3 hours. Diluting with water (50 mL), extracting with ethyl acetate (80 mL), concentrating the organic phase, and purifying with flash silica gel column to obtain the compound 9-3.MS m/z(ESI):197.8[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.41(d,J＝3.30Hz,1H),6.51(d,J＝3.52Hz,1H),4.04(s,3H),3.75(s,3H).

Step 3: synthesis of Compound 9-4

A-1 (1.88 g,14.6 mmol) and Compound 9-3 (2.50 g,12.7 mmol) were dissolved in anhydrous dioxane (50 mL), cesium carbonate (12.4 g,38.0 mmol), 4, 5-bis (diphenylphosphorus) -9, 9-dimethylxanthene (1.46 g,2.53 mmol), tris (dibenzylideneacetone) dipalladium (2.32 g,2.53 mmol) were added and the nitrogen was replaced three times and reacted at 110℃for 18 hours. Diluting with water (200 mL), extracting with ethyl acetate (200 mL), concentrating the organic phase, and purifying with flash silica gel column to obtain compound 9-4.MS m/z(ESI):291.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.14(d,J＝3.46Hz,1H),6.37(d,J＝3.46Hz,1H),5.26(q,J＝5.90Hz,1H),3.98(s,3H),3.67(s,3H),2.96(t,J＝7.03Hz,1H),2.66–2.58(m,1H),2.40(s,3H),2.20–2.13(m,1H),1.77–1.62(m,4H),1.25(d,J＝6.32Hz,3H).

Step 4: synthesis of Compound 9-5

Compound 9-4 was dissolved in (720 mg,2.48 mmol) and acetonitrile (15 mL), and after adding trimethyliodosilane (2.48 g,12.4 mmol), it was reacted at 25℃for 1 hour, and then heated to 80℃for 3 hours. The reaction solution is dried and concentrated and then purified by a rapid silica gel column to obtain the compound 9-5.MS m/z(ESI):276.9[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:11.90(br s,1H),7.00-6.86(m,1H),6.36(d,J＝3.08Hz,1H),5.20-4.90(m,1H),3.98-3.85(m,3H),3.63(s,3H),3.59-3.47(m,1H),3.01(d,J＝4.40Hz,2H),2.30-2.17(m,1H),2.15–2.00(m,1H),1.98–1.75(m,2H),1.44(d,J＝6.16Hz,3H).

Step 5: synthesis of Compound 9-6

Compound 9-5 (600 mg,2.17 mmol) was dissolved in toluene (15 mL), phosphorus oxychloride (3.33 g,21.7 mmol) and N, N-diisopropylethylamine (560 mg,4.34 mmol) were added, and then reacted at 80℃for 3 hours. The solvent was removed under reduced pressure, the remaining oil was poured into ice water (20 mL), pH 8-9 was adjusted using saturated sodium bicarbonate solution, extracted with dichloromethane (20 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give compound 9-6.MS m/z (ESI) 294.9[ M+H ] ⁺.

Step 6: synthesis of Compound 9-7

Compound 9-6 (480 mg,1.63 mmol) was dissolved in dimethyl sulfoxide (6 mL), triethylenediamine (182 mg,1.63 mmol) was added and stirred at 25℃for 1 hour, and sodium cyanide (150 mg,3.06 mmol) was added and reacted at 80℃for 2 hours. Water (20 mL) was added to dilute, extraction was performed with ethyl acetate (20 mL), the aqueous phase was further extracted with ethyl acetate (10 mL. Times.3), the combined organic phases were washed with brine (50 mL), and the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give Compound 9-7.MS m/z (ESI): 286.0[ M+H ] ⁺.

Step 7: synthesis of Compound 9-8

Compound 9-7 (150 mg,0.53 mmol) was dissolved in absolute ethanol (3 mL), hydroxylamine hydrochloride (95 mg,1.37 mmol) and sodium carbonate (72.4 mg,0.68 mmol) were added and reacted at 85℃for 2 hours. Water (10 mL) was added for dilution, extraction was performed with ethyl acetate (10 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give compound 9-8.MS m/z (ESI): 319.0[ M+H ] ⁺.

Step 8: synthesis of Compound 9-9

Compound 9-8 (140 mg,0.44 mmol) and compound A-2 (114 mg,0.48 mmol) were dissolved in N, N-dimethylformamide (3 mL), N-diisopropylethylamine (171 mg,1.32 mmol) was added dropwise, then cooled to 0℃and stirred for 0.5 hours, benzotriazol-1-yl-oxy-tripyrrolidinylphosphine hexafluorophosphate (349 mg,0.66 mmol) was added, and then stirred for 5.5 hours at 25 ℃. Water (10 mL) was added for dilution, extraction was performed with ethyl acetate (10 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give compound 9-9.MS m/z (ESI): 537.2[ M+H ] ⁺.

Step 9: preparation of Compound 9

Compound 9-9 (266 mg,0.50 mmol) was dissolved in N, N-dimethylformamide (4 mL), and anhydrous sodium carbonate (158 mg,1.49 mmol) was added and reacted at 100℃for 48 hours. Filtering, purifying with preparative high performance liquid chromatograph (chromatographic column: boston Prime C18,150 ×30mm,5 μm; mobile phase: 0.05% ammonia water solution (wt 0.08% ammonium bicarbonate) -acetonitrile; gradient: 53% -73%,10 min), to obtain compound 9.MS m/z(ESI):541.0[M+Na]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.55(d,J＝3.60Hz,1H),7.11(s,2H),6.87(d,J＝3.60Hz,1H),5.45-5.26(m,1H),3.77(s,3H),3.04-2.92(m,1H),2.71-2.65(m,1H),2.62-2.53(m,2H),2.43(s,3H),2.20-2.11(m,2H),2.02-1.95(m,1H),1.91-1.82(m,5H),1.82-1.74(m,2H),1.71-1.63(m,2H),1.29(d,J＝6.40Hz,3H).

Example 10: preparation of Compound 10

Step 1: synthesis of Compound 10-2

Compound 8-3 (800 mg,4.17 mmol), A-1 (539.4 mg,4.17 mmol), sodium tert-butoxide (802.4 mg,8.35 mmol), dioxane (10 mL) were added to the reaction flask. After purging with nitrogen, chloro (2-dicyclohexylphosphine-2 ',4',6 '-triisopropylbiphenyl) (2' -aminobiphenyl-2-yl) palladium (II) methanesulfonate (188.8 mg,0.21 mmol) was added. After nitrogen substitution, the reaction mixture was stirred at 80℃for 12 hours. The reaction was cooled to room temperature and quenched with water (15 mL). The aqueous phase was extracted with ethyl acetate (30 mL. Times.2), the organic phases were combined and washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude product, which was purified by C18 reverse phase silica gel chromatography (acetonitrile/water (0.1% trifluoroacetic acid): 5-35%) to give 10-2.MS m/z (ESI): 285.3[ M+H ] ⁺.

Step 2: synthesis of Compound 10-3

10-2 (80 Mg,0.28 mmol), hydroxylamine hydrochloride (19.6 mg,0.28 mmol), sodium carbonate (29.8 mg,0.28 mmol) and ethanol (3 mL) were added to the reaction flask, and the mixture was heated to 80℃for 2 hours. The reaction solution was cooled to room temperature, filtered, and the cake was washed with dichloromethane (30 mL), and the filtrate was collected and concentrated to give product 10-3.MS m/z (ESI): 318.2[ M+H ] ⁺.

Step 3: synthesis of Compound 10-4

2- (7-Azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (115.01 mg,0.30 mmol) was added to a solution of A-2 (59.6 mg,0.25 mmol), 10-3 (80 mg,0.25 mmol), N, N-diisopropylethylamine (0.13 mL,0.76 mmol) in N, N-dimethylformamide (2 mL). Stirred at 25℃for 12 hours. The reaction was added dropwise to an ice ammonium chloride solution (15 mL), and the aqueous phase was extracted with ethyl acetate (3×50 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated. Purification of the crude product by C18 reverse phase silica gel chromatography (acetonitrile/water (0.1% trifluoroacetic acid): 5-38%) gives 10-4.MS m/z (ESI): 536.4[ M+H ] ⁺.

Step 4: synthesis of Compound 10

Compound 10-4 (70 mg,0.13 mmol), sodium carbonate (69.3 mg,0.65 mmol), dimethyl sulfoxide (0.5 mL), dioxane (0.5 mL) were added to the reaction flask, and the reaction mixture was stirred at 100deg.C for 12 hours. The reaction mixture was cooled to room temperature and filtered. The filtrate was diluted with ethyl acetate (50 mL), and washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and concentrated by filtration to give a crude product. Purifying the crude product with preparative high performance liquid chromatography column (chromatography column: xtimate C18,150 X10 mm,5 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 20% -50%,12.5 min) to obtain compound 10 (formate) ).MS m/z(ESI):518.0[M+H]⁺.¹H NMR(400MHz,Methanol-d₄)δ:8.55(s,1H),7.39(s,1H),7.21-7.05(m,2H),5.14-4.94(m,1H),3.87-3.77(m,3H),3.62-3.46(m,1H),2.83-2.71(m,3H),2.70-2.52(m,3H),2.30-2.19(m,2H),2.17-1.74(m,10H),1.44(d,J＝6.4Hz,3H).

Examples 11,12: preparation of Compounds 11,11a,11b,12

Step1: synthesis of Compound 11-1:

compound A-4 (10.0 g,73.4 mmol) was dissolved in ethanol (100 mL), sodium borohydride (2.85 g,75.3 mmol) was added at 0deg.C, warmed to room temperature, and stirred for 3 hours. After the completion of the reaction, the reaction mixture was poured into 100mL of saturated ammonium chloride, quenched, concentrated under reduced pressure, and extracted with ethyl acetate (100 mL. Times.3). The combined organic phases were washed with saturated brine (70 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to give the crude product. Purifying the crude product by silica gel column chromatography to obtain the compound 11-1.MS m/z(ESI):139.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.82(dd,J＝4.8,2.0Hz,1H),7.39(dd,J＝7.2,2.0Hz,1H),6.53(dd,J＝7.2,4.8Hz,1H),5.70(s,2H),5.25(d,J＝4.0Hz,1H),4.71(qd,J＝6.4,2.8Hz,1H),1.31(d,J＝6.4Hz,3H).

Step 2: synthesis of Compound 11-2:

Compound 11-1 (9.5 g,68.8 mmol) was dissolved in tetrahydrofuran (100 mL), thionyl chloride (16.4 g,137.7 mmol) was added at 0deg.C, warmed to room temperature, and stirred for 18 hours. After the reaction is finished, directly decompressing and concentrating the reaction solution to obtain the compound 11-2.¹H NMR(400MHz,DMSO-d₆)δ:8.28(s,2H),8.22(dd,J＝7.6,1.6Hz,1H),8.05(dd,J＝6.4,1.6Hz,1H),6.96(dd,J＝7.6,6.4Hz,1H),5.52(q,J＝6.8Hz,1H),1.84(d,J＝6.8Hz,3H).

Step 3: synthesis of Compound 11-3:

Compound 11-2 (5.5 g,35.1 mmol) was dissolved in acetic anhydride (50 mL), warmed to 100deg.C and stirred for 18 hours. After the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product. Purifying the crude product by silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain the compound 11-3.MS m/z(ESI):241.2[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.57(dd,J＝4.8,1.8Hz,1H),8.29(dd,J＝8.0,1.6Hz,1H),7.64(dd,J＝8.0,4.8Hz,1H),5.31(q,J＝6.8Hz,1H),2.40(s,3H),1.99(s,3H),1.75(d,J＝6.8Hz,3H).

Step 4: synthesis of Compound 11-4:

Compound A-5 (1.7 g,8.51 mmol) was dissolved in N, N-dimethylformamide (25 mL), sodium hydride (623 mg,15.6mmol, purity 60%) was added at 0deg.C, stirred for 30 minutes, then compound 11-3 (2.5 g,10.4 mmol) was added, and the mixture was stirred for 3 hours at 50deg.C. After the reaction, the reaction mixture was concentrated, quenched in 50mL of saturated ammonium chloride, and extracted with ethyl acetate (70 mL. Times.3). The organic phases were combined, washed with saturated brine (70 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to give the compound 11-4.MS m/z(ESI):362.2[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:10.17(s,1H),8.54(dd,J＝4.8,1.6Hz,1H),8.29(dd,J＝7.6,1.6Hz,1H),7.84(d,J＝3.6Hz,1H),7.53(dd,J＝7.6,4.8Hz,1H),6.74(d,J＝3.6Hz,1H),6.36(q,J＝7.2Hz,1H),2.75(s,3H),2.17(s,3H),2.05(d,J＝7.2Hz,3H).

Step 5: synthesis of Compound 11-5:

compound 11-4 (2.5 g,6.9 mmol) and zinc cyanide (1.17 g,10.0 mmol) were dissolved in N-methylpyrrolidone (25 mL) and ditri-butylphosphine palladium (100 mg,0.2 mmol) was added under nitrogen. The reaction solution was placed in a microwave and heated to 160℃and stirred for 1 hour. After completion of the reaction, the reaction mixture was poured into 20mL of water, and extracted with ethyl acetate (55 mL. Times.2). The organic phases were combined, washed with saturated brine (30 mL. Times.5), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product. Purifying the crude product by silica gel column chromatography to obtain the compound 11-5.MS m/z(ESI):353.2[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:9.98(s,1H),8.38(dd,J＝4.8,1.6Hz,1H),8.15(dd,J＝7.8,1.6Hz,1H),7.92(d,J＝3.6Hz,1H),7.37(dd,J＝7.6,4.8Hz,1H),6.77(d,J＝3.6Hz,1H),6.23(q,J＝7.2Hz,1H),2.60(s,3H),1.98(s,3H),1.89(d,J＝7.2Hz,3H).

Step 6: synthesis of Compound 11-6:

Compound 11-5 (1.75 g,5.0 mmol) was dissolved in ethanol (30 mL) and sodium carbonate (1.06 g,10.0 mmol) and hydroxylamine hydrochloride (690 mg,10.0 mmol) were added. The reaction solution was raised to 90℃and stirred for 16 hours. Filtering sodium carbonate, washing the filter cake with dichloromethane (30 mL) and methanol (10 mL) for 3 times, mixing the organic phases, concentrating, pulping with water (15 mL) at room temperature for 0.5 hr, and filtering to obtain the compound 11-6.MS m/z(ESI):344.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:10.41(s,1H),7.87(dd,J＝4.8,1.6Hz,1H),7.66(dd,J＝3.6,1.2Hz,1H),7.37(dd,J＝7.6,1.6Hz,1H),6.86(d,J＝3.6Hz,1H),6.54(dd,J＝7.6,4.8Hz,1H),6.02(q,J＝6.8Hz,1H),5.95(s,2H),5.87(s,2H),2.60(s,3H),1.81(d,J＝7.2Hz,3H).

Step 7: synthesis of Compound 11-7:

compound 11-6 (730 mg,2.13 mmol) and compound A-2-P1 (503 mg,2.13 mmol) were dissolved in N, N-dimethylformamide (10 mL), N-diisopropylethylamine (827 mg,6.40 mmol) was added dropwise, and after cooling to 0℃and stirring for 0.5 hours, benzotriazol-1-yl-oxy-tripyrrolidinylphosphine hexafluorophosphate (1.66 g,3.20 mmol) was added, and then stirred for 18 hours at 25 ℃. The reaction mixture was poured into water (20 mL), ethyl acetate (40 mL. Times.3) and extracted, and the combined organic phases were concentrated and purified using a flash silica gel column to give the compound 11-7.MS m/z(ESI):562.2[M+H]⁺.¹H NMR(400MHz,Methanol-d₄)δ:7.77(dt,J＝5.2,1.6Hz,1H),7.56–7.50(m,1H),7.35(d,J＝4.0Hz,1H),7.02(d,J＝3.6Hz,1H),6.59(dd,J＝7.6,5.2Hz,1H),5.99(q,J＝7.2Hz,1H),2.53(s,3H),2.46(td,J＝6.0,3.6Hz,2H),1.82(d,J＝5.6Hz,2H),1.78(d,J＝7.2Hz,3H),1.67(s,2H),1.62(d,J＝2.0Hz,3H).

Step 8: synthesis of Compound 11-8:

Compound 11-7 (1.05 g,1.87 mmol) and sodium carbonate (640 mg,6.08 mmol) were dissolved in dimethyl sulfoxide (10 mL), and the reaction was heated to 120℃and stirred for 16 h. After the reaction was completed, water (10 mL) was added to the reaction mixture, extracted with ethyl acetate (30 mL. Times.3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated to give a crude product, and the crude product was purified by flash silica gel column to give the compound 11-8.MS m/z(ESI):544.3[M+H]⁺.¹H NMR(400MHz,CDCl₃)δ:8.00–7.92(m,1H),7.61(dd,J＝7.6,1.6Hz,1H),7.05–6.91(m,2H),6.71–6.61(m,1H),6.03(q,J＝7.2Hz,1H),2.63(s,3H),2.61–2.51(m,2H),1.92–1.82(m,7H),1.79(d,J＝7.2Hz,3H).

Step 9: example 12 and synthesis of compounds 11-9:

Compound 11-8 (530 mg,0.97 mmol) was dissolved in tetrahydrofuran (10 mL) and water (5 mL), and potassium peroxymonosulfonate (540 mg,0.88 mmol) was added and stirred at room temperature for 1 hour. After the completion of the reaction, water (5 mL) was added to the reaction mixture, extracted with ethyl acetate (10 mL. Times.3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by a flash silica gel column (eluent: methanol/dichloromethane) to give compound 11-9 and compound 12. Compound 12 can be further purified by high performance liquid chromatography (column: waters-SunFire-C18, 19X 250mm,10 μm; mobile phase: 0.1% aqueous formic acid-acetonitrile; gradient: acetonitrile 11% -41%,8 min). Compounds 11-9: MS m/z (ESI): 560.3[ M+H ] ⁺.

Compounds of formula (I) 12：MS m/z(ESI):576.3[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.32(dd,J＝3.6,2.4Hz,1H),7.90(dt,J＝4.8,1.6Hz,1H),7.44–7.41(m,1H),7.23(d,J＝3.6Hz,1H),7.10(s,2H),6.58–6.54(m,1H),6.22(q,J＝6.8Hz,1H),5.98(s,2H),3.47(s,3H),2.71–2.55(m,2H),2.22–2.13(m,1H),2.05–1.97(m,1H),1.95–1.79(m,8H).

Step 10: synthesis of Compounds 11-10:

Compound 11-9 (153 mg,0.27 mmol) was dissolved in N, N-dimethylformamide (3 mL), and N, N-dimethylformamide dimethyl acetal (326 mg,2.7 mmol) was added thereto and stirred at room temperature for 18 hours. Water (10 mL) was added to the reaction solution, extracted with methylene chloride (10 mL. Times.3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product, which was purified by flash silica gel column to give compound 11-10.MS m/z (ESI): 615.4[ M+H ] ⁺.

Step 11: synthesis of Compounds 11-11:

Compound 11-10 (120 mg,0.20 mmol) was dissolved in tetrahydrofuran (1 mL), compound A-1 (78 mg,0.60 mmol) was added to the system, cooled to 0deg.C, sodium t-butoxide (48 mg,0.50 mmol) was added, and stirred for 30 min. After the completion of the reaction, the reaction mixture was poured into 5mL of saturated ammonium chloride, quenched, and extracted with ethyl acetate (10 mL. Times.2). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated to dryness under reduced pressure, and the residue was purified by silica gel column chromatography to give compound 11-11.MS m/z (ESI): 680.5[ M+H ] ⁺.

Step 12: preparation of Compound 11:

compound 11-11 (100 mg,0.15 mmol) was dissolved in ethanol (3 mL), concentrated hydrochloric acid (90. Mu.L) was added, and the reaction solution was heated to 100℃and stirred for 60 minutes. After the reaction is finished, crude products are obtained, and the crude products are purified by high performance liquid chromatography (chromatographic column: xtimate C18,150 multiplied by 40mm,5 mu m; mobile phase: 0.1% ammonium bicarbonate aqueous solution-acetonitrile; gradient: acetonitrile 35% -65%,15 min) to obtain the compounds 11.MS m/z(ESI):625.8[M+H]⁺.¹HNMR(400MHz,DMSO-d₆)δ:7.87(dt,J＝4.8,2.4Hz,1H),7.74(dd,J＝7.2,3.6Hz,1H),7.25(dd,J＝47.6,7.6Hz,1H),7.09(s,2H),6.92(dd,J＝6.4,3.6Hz,1H),6.52(td,J＝7.2,4.8Hz,1H),5.98(dd,J＝7.2,4.0Hz,1H),5.90(s,2H),5.29(dt,J＝19.6,5.6Hz,1H),2.95(s,1H),2.70–2.54(m,3H),2.41(s,1H),2.34(s,2H),2.21–2.08(m,2H),2.02–1.94(m,1H),1.92–1.73(m,10H),1.66(s,2H),1.27(d,J＝6.4Hz,2H),1.21(d,J＝6.4Hz,1H).

Step 13: synthesis of example 11a and example 11b

Example 11 (80 mg,0.15 mmol) was separated by preparative supercritical fluid chromatography (column: CHIRALPAK AD,250 x 30mm i.d.,10 μm, mobile phase: carbon dioxide-isopropanol (0.1% ammonia; gradient: isopropanol 40%, flow rate: 130 mL/min) to give crude 11a (retention time 2.750 min) and 11b (retention time 7.714 min), crude 11a and 11b were purified by preparative high performance liquid chromatography (column: xtimate C18,150 x 40mm,5 μm; mobile phase: 0.1% ammonium bicarbonate aqueous solution-acetonitrile, gradient: acetonitrile 35% -100%,17 min) to give the corresponding product compound 11a, compound 11b, respectively;

compounds of formula (I) 11a：MS m/z(ESI):625.8[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.86(dd,J＝4.8,1.6Hz,1H),7.77(s,1H),7.17–7.13(m,1H),7.12(s,2H),6.93(d,J＝3.6Hz,1H),6.51(dd,J＝7.2,4.8Hz,1H),5.99–5.96(m,1H),5.93(s,2H),5.36–5.27(m,1H),2.93–2.91(m,1H),2.70–2.63(m,2H),2.37–2.29(m,4H),2.19–2.06(m,2H),2.01–1.60(m,13H),1.27–1.25(m,3H).

Compounds of formula (I) 11b：MS m/z(ESI):625.8[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.87(dd,J＝4.8,1.6Hz,1H),7.73(d,J＝3.6Hz,1H),7.37–7.28(m,1H),7.09(s,2H),6.91(d,J＝3.6Hz,1H),6.53(dd,J＝7.6,4.8Hz,1H),5.97(dd,J＝6.8Hz,1H),5.89(s,2H),5.27–5.24(m,1H),2.96–2.94(m,1H),2.69–2.54(m,3H),2.40(s,3H),2.21–2.10(m,2H),2.02–1.63(m,13H),1.20(d,J＝6.4Hz,3H).

Example 13: preparation of Compound 13

Step 1: synthesis of Compound 13-2

To a solution of compound 13-1 (35 g,281.94 mmol) in tetrahydrofuran (500 mL) was slowly added dropwise thionyl chloride (50 mL,689.31 mmol) at 0deg.C. After the dripping, the reaction liquid is naturally heated to 25 ℃ for reaction for 3 hours. After the reaction is finished, a large amount of solid is separated out from the reaction solution, the reaction solution is filtered, a filter cake is leached by diethyl ether (50 mL), and the filter cake is dried to obtain the compound 13-2 (hydrochloride) ).¹H NMR(400MHz,DMSO-d6)δ:14.36(br,1H),8.31(s,2H),8.12(dd,J＝7.4,1.7Hz,1H),8.05(dd,J＝6.3,1.7Hz,1H),6.92(dd,J＝7.3,6.3Hz,1H),4.87(s,2H).

Step 2: synthesis of Compound 13-3

The hydrochloride salt of compound 13-2 (19 g,106.12 mmol) was added to acetic anhydride (150 mL), and the reaction mixture was heated to 100deg.C for 24 hours. Concentrating the reaction solution, and purifying by using a rapid silica gel column to obtain a compound 13-3.MS m/z(ESI):227.0[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ8.65–8.54(m,1H),8.20–8.09(m,1H),7.65–7.55(m,1H),4.69(s,2H),2.20(s,6H).

Step 3: synthesis of Compound 13-4

Sodium hydride (928 mg,23.2mmol, 60%) was added slowly to A-5 (3.86 g,19.3 mmol) and N, N-dimethylformamide (80 mL) at 0deg.C. After the completion of the addition, the reaction mixture was stirred for half an hour, 13-3 (4.38 g,19.3 mmol) was further added thereto, and the reaction mixture was heated to 25℃for 1 hour. Saturated ammonium chloride solution (100 mL) was added to quench and diluted with water (500 mL), extracted with ethyl acetate (50 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and purified using flash silica gel column to give compound 13-4.MS m/z (ESI): 390.1[ M+H ] ⁺.

Step 4: synthesis of Compound 13-5

13-4 (2 G,5.13 mmol), zinc cyanide (0.90 g,7.70 mmol), bis-tri-tert-butylphosphine palladium dichloride (0.26 g,0.51 mmol), N-methylpyrrolidone (15 mL) were added to a 20 mL microwave tube, after purging nitrogen. The reaction was carried out in a microwave reactor at 110℃for 1 hour. The reaction solution was filtered, diluted with ethyl acetate (100 mL), washed with saturated brine (30 ml×6), dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by normal phase silica gel chromatography to give 13-5.MS m/z (ESI): 339.1[ M+H ] ⁺.

Step 5: synthesis of Compound 13-6

13-5 (920 Mg,2.72 mmol), hydroxylamine hydrochloride (188.9 mg,2.72 mmol), sodium carbonate (1440.8 mg,13.59 mmol) were added to ethanol (20 mL) and reacted at 80℃for 1 hour. The reaction solution was cooled to room temperature, filtered, and the cake was washed with dichloromethane (50 mL), and the filtrate was collected and concentrated to give compound 13-6.MS m/z (ESI): 372.3[ M+H ] ⁺.

Step 6: synthesis of Compound 13-7

Tripyrrolidinyl phosphonium bromide hexafluorophosphate (1.45 g,2.79 mmol) was added to a solution of 13-6 (690.0 mg,1.86 mmol), A-2-P1 (439.5 mg,1.86 mmol), N-diisopropylethylamine (923. Mu.L, 5.57 mmol) in N, N-dimethylformamide (15 mL). Stirred at 25℃for 12 hours. The reaction was added dropwise to a saturated ammonium chloride solution of ice (30 mL) and extracted with ethyl acetate (50 ml×3). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and the concentrated crude product was purified by flash chromatography on silica gel to give compound 13-7.MS m/z (ESI): 590.2[ M+H ] ⁺.

Step 7: synthesis of Compound 13-8

Compound 13-7 (720 mg,1.22 mmol), sodium carbonate (646.5 mg,6.10 mmol), dimethyl sulfoxide (4 mL) were added to dioxane (4 mL), and the reaction mixture was stirred at 100deg.C for 12 hr. The reaction mixture was cooled to room temperature and filtered. The filtrate was diluted with ethyl acetate (100 mL) and washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, concentrated by filtration, and the residue was purified by flash chromatography on silica gel to give the compound 13-8.MS m/z(ESI):572.3[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:10.28(s,1H),8.37-8.33(m,1H),7.66-7.58(m,1H),7.40-7.33(m,1H),7.24-7.19(m,1H),7.17-7.01(m,2H),6.99-6.93(m,1H),5.42(s,2H),2.57-2.54(m,3H),2.20-2.04(m,5H),2.01-1.95(m,2H),1.95-1.70(m,5H).

Step 8: synthesis of Compound 13-9

Potassium peroxomonosulphonate (400 mg,0.65 mmol) was added to a mixed solution of 13-8 (580 mg,1.01 mmol) in tetrahydrofuran (10 mL) and water (4 mL), and the mixture was stirred at 50℃for 2 hours. The reaction mixture was quenched with saturated aqueous sodium sulfite (4 mL), saturated ammonium chloride (10 mL), and the aqueous phase was extracted with ethyl acetate (50 mL. Times.3). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. Purifying the crude product by a rapid silica gel chromatographic column to obtain 13-9.MS m/z(ESI):588.2[M+H]⁺.¹HNMR(400MHz,DMSO-d₆)δ:10.33(s,1H),8.39-8.32(m,1H),8.02-7.94(m,1H),7.35-7.31(m,1H),7.23-7.18(m,1H),7.18-7.15(m,1H),7.14-7.04(m,2H),5.53(s,2H),2.93-2.86(m,3H),2.61-2.56(m,1H),2.30-2.06(m,5H),2.06-1.80(m,6H).

Step 9: synthesis of Compound 13-10

Sodium tert-butoxide (69.5 mg,0.72 mmol) was added to a solution of 13-9 (85 mg,0.14 mmol) and A-1 (93.44 mg,0.72 mmol) in tetrahydrofuran (2 mL) at 0deg.C. The reaction was carried out at 0℃for 1.5 hours. The reaction was added dropwise to a saturated ammonium chloride solution of ice (20 mL) and extracted with ethyl acetate (50 mL. Times.3). The obtained organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 13-10.MS m/z (ESI): 653.4[ M+H ] ⁺.

Step 10: preparation of Compound 13

1M aqueous hydrochloric acid (1 mL) was added dropwise to a solution of 13-10 (60 mg,0.09 mmol) in ethanol (0.5 mL), and the mixture was reacted at 90℃for 2 hours. The reaction was added dropwise to a saturated sodium bicarbonate solution of ice (15 mL) and extracted with ethyl acetate (50 ml×3). The obtained organic phase is dried with anhydrous sodium sulfate, filtered, concentrated, and the residue is purified by preparative high performance liquid chromatography (column: xtimate C18,150 ×40mm,5 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 20% -25%,15 min) to obtain the compound 13.MS m/z(ESI):611.7[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.19(s,1H),7.93–7.81(m,1H),7.64(d,J＝3.6Hz,1H),7.10–6.89(m,3H),6.47(dd,J＝7.2,4.8Hz,1H),6.04(s,2H),5.36–5.27(m,1H),5.26–5.10(m,2H),2.97–2.92(m,1H),2.66–2.61(m,1H),2.60–2.53(m,2H),2.21–2.10(m,2H),2.08–2.06(m,4H),2.02–1.88(m,2H),1.84(s,3H),1.81–1.72(m,2H),1.71–1.57(m,2H),1.25(d,J＝6.4Hz,3H).

Example 14: preparation of Compound 14

Step1: synthesis of Compound 14-2

Sodium tert-butoxide (65.4 mg,0.68 mmol) was added to a solution of 13-9 (80 mg,0.14 mmol), 14-1 (108.3 mg,0.68 mmol) in tetrahydrofuran (2 mL) at 0deg.C. The reaction was carried out at 0℃for 10 minutes. The reaction was added dropwise to a saturated ammonium chloride solution of ice (20 mL) and extracted with ethyl acetate (50 mL. Times.3). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 14-2.MS m/z (ESI): 683.3[ M+H ] ⁺.

Step 2: synthesis of example 14

1M aqueous hydrochloric acid (1 mL) was added dropwise to a solution of 14-2 (70 mg,0.11 mmol) in ethanol (1 mL), and the mixture was reacted at 90℃for 2 hours. The reaction was added dropwise to a saturated sodium bicarbonate solution (15 mL) of ice, and extracted with ethyl acetate (50 mL. Times.3). The obtained organic phase is dried with anhydrous sodium sulfate, filtered, concentrated, and the residue is purified by preparative high performance liquid chromatography (column: xtimate C18,150X 40mm,5 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 20% -25%,15 min) to obtain compound 14 (formate salt) ).MS m/z(ESI):641.1[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.18(s,1H),7.90-7.82(m,1H),7.64(d,J＝3.6Hz,1H),7.10(s,2H),6.99-6.86(m,2H),6.47(dd,J＝7.2,4.8Hz,1H),6.08(s,2H),5.36-5.19(m,3H),4.17-4.04(m,2H),3.13-3.02(m,3H),2.86-2.80(m,1H),2.58-2.55(m,1H),2.22-2.11(m,2H),2.10-2.06(m,1H),2.06-1.89(m,3H),1.89-1.52(m,8H).

Example 15: preparation of Compound 15

Step1: synthesis of Compound 15-2

Sodium tert-butoxide (53.2 mg,0.55 mmol) was added to a solution of 15-1 (71.5 mg,0.55 mmol), 13-9 (65 mg,0.11 mmol) in tetrahydrofuran (2 mL) at 0deg.C. The reaction was carried out at 0℃for 2 hours. The reaction was added dropwise to a saturated ammonium chloride solution of ice (20 mL) and extracted with ethyl acetate (50 mL. Times.3). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 15-2.MS m/z (ESI): 653.3[ M+H ] ⁺.

Step 2: synthesis of example 15

1M aqueous hydrochloric acid (1 mL) was added dropwise to a solution of 15-2 (50 mg,0.08 mmol) in ethanol (1 mL), and the mixture was reacted at 90℃for 5 hours. The reaction was added dropwise to a saturated sodium bicarbonate solution of ice (15 mL) and extracted with ethyl acetate (50 ml×3). The resulting organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. Purifying the crude product with preparative high performance liquid chromatography column (chromatography column: xtimate C18,150 X10 mm,5 μm; mobile phase: 0.225% formic acid water solution-acetonitrile; gradient: 20% -25% acetonitrile, 15 min) to obtain compound 15 (formate) ).MS m/z(ESI):611.0[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:8.28(s,1H),7.92-7.82(m,1H),7.63(d,J＝3.6Hz,1H),7.10(s,2H),6.99-6.83(m,2H),6.51-6.41(m,1H),6.08(s,2H),5.24(s,2H),4.26(s,2H),2.64-2.55(m,2H),2.31-2.24(m,2H),2.17(s,6H),2.13-1.86(m,4H),1.85(s,3H),0.74-0.57(m,2H),0.48-0.32(m,2H).

Example 16: preparation of Compound 16, compound 16a, compound 16b

Step1: synthesis of Compound 16-2

To a solution of compounds 1 to 12 (800 mg,2.20 mmol) in ethanol (10 mL) was added aqueous sodium hydroxide (10 mL, 4M), and the reaction was stirred at 90℃for 12 hours. The pH of the reaction solution is regulated to 6-7 by dilute hydrochloric acid, the reaction solution is concentrated, and the crude product is purified by rapid silica gel column chromatography (eluent: dichloromethane/methanol) to obtain the compound 16-2.MS m/z (ESI): 384.2[ M+H ] ⁺.

Step 2: synthesis of Compound 16-3

To a solution of compound 16-2 (720 mg,1.88 mmol), 2- (7-azabenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate (1.07 g,2.82 mmol) and N, N-diisopropylethylamine (0.93 mL,5.63 mmol) in N, N-dimethylformamide (10 mL) was added dimethylhydroxylamine hydrochloride (550.13 mg,5.64 mmol), and the reaction mixture was stirred at 50℃for 12 hours. The reaction solution was concentrated and the crude product was purified by flash column chromatography on silica gel (eluent: dichloromethane/methanol) to give compound 16-3.MS m/z (ESI): 427.2[ M+H ] ⁺.

¹H NMR(400MHz,DMSO-d₆)δ:7.85(d,J＝4.0Hz,1H),7.61(s,1H),6.90(s,1H),6.79(d,J＝4.0Hz,1H),5.42-5.35(m,1H),4.08(s,3H),3.94(s,3H),3.76-3.73(m,4H),3.57-3.53(m,1H),3.32(s,3H),3.19-3.13(m,1H),3.05(d,J＝8.0Hz,3H),2.33-2.26(m,1H),2.12-2.06(m,1H),1.95-1.84(m,2H),1.44(d,J＝4.0Hz,3H).

Step 3: synthesis of Compound 16-4

To a solution of Compound A-2 (2.6 g,11.00 mmol) in tetrahydrofuran (50 mL) was added N, N' -carbonyldiimidazole (2.68 g,16.51 mmol), and the reaction was stirred at 50℃for 1 hour. Cooled to 25℃and sodium borohydride (1.25 g,33.01 mmol) and water (625. Mu.L) were added. The reaction solution was stirred at 25℃for 1 hour. To the reaction mixture was added water (50 mL), which was quenched, extracted with ethyl acetate (50 mL. Times.2), and the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (ethyl acetate/petroleum ether) to give compound 16-4.MS m/z (ESI): 223.0[ M+H ] ⁺.

Step 4: synthesis of Compound 16-5

N, N-dimethylformamide dimethyl acetal (3.43 mL,25.64 mmol) was added to compound 16-4 (950 mg,4.27 mmol), and the reaction mixture was stirred in an ultrasonic water bath at 25℃for 15 minutes to give a clear solution. Water (20 mL) was added to the reaction solution, and stirring was continued in an ultrasonic water bath until suspended. The solid was collected by filtration. The solid was washed with water and concentrated under reduced pressure to give compound 16-5.MS m/z (ESI): 278.2[ M+H ] ⁺.

Step 5: synthesis of Compound 16-6

To a solution of oxalyl chloride (0.70 mL,8.11 mmol) in dichloromethane (10 mL) was added dropwise a solution of dimethyl sulfoxide (1.04 mL,14.60 mmol) in dichloromethane (10 mL) under nitrogen at-78deg.C for 30 min, then compound 16-5 (900 mg,3.24 mmol) was slowly added, stirred at-78deg.C for 30 min, and triethylamine (4.05 mL,29.20 mmol) was added dropwise. The reaction solution was heated to 10℃and stirred under nitrogen atmosphere for 2 hours. The reaction was diluted with water (20 mL) and extracted with dichloromethane (20 mL. Times.3). The organic phase was washed with saturated brine (20 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain the compound 16-6.MS m/z (ESI): 276.2[ M+H ] ⁺.

Step 6: synthesis of Compound 16-7

To a methanol solution (15 mL) of compound 16-6 (720 mg,2.61 mmol) and cesium carbonate (1.02 g,3.14 mmol) was added a methanol solution (5 mL) of dimethyl (1-diazo-2-oxopropyl) phosphonate (552.53 mg,2.88 mmol) at 0℃and the reaction solution was stirred at 0℃for 2 hours. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (20 mL. Times.3). The organic phase was washed with saturated brine (10 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by flash silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain the compound 16-7.MS m/z (ESI): 272.1[ M+H ] ⁺.

Step 7: synthesis of Compound 16-8

To a solution of compound 16-7 (222.69 mg,0.82 mmol) in tetrahydrofuran (5 mL) was added lithium bis trimethylsilylamide (2.46 mL,2.46 mmol) under nitrogen at-78deg.C, followed by compound 16-3 (350 mg,0.82 mmol). The reaction solution was warmed to 10℃and stirred under nitrogen for 30 minutes. The reaction mixture was quenched with water (10 mL) and extracted with ethyl acetate (20 mL. Times.3). The organic phase was washed with saturated brine (10 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and the crude product was purified by flash column chromatography on silica gel (eluent: dichloromethane/methanol) to give compound 16-8.MS m/z (ESI): 637.4

[M+H]⁺。

Step 8: synthesis of Compound 16-9

A methanol mixed solution (5 mL) of compound 16-8 (400 mg,0.63 mmol) and hydroxylamine hydrochloride (218.24 mg,3.14 mmol) was stirred at 50℃for 12 hours. The reaction solution was concentrated to obtain compound 16-9.MS m/z (ESI): 652.4[ M+H ] ⁺.

Step 9: synthesis of Compound 16

A solution of compound 16-9 (300 mg,0.46 mmol) in hydrogen chloride dioxane (3 mL, 4M) was stirred at 50℃for 2 hours. The reaction solution was concentrated to obtain a crude product. Purification by preparative high performance liquid chromatography (column: boston Prime C18,150X 30mm,5 μm; mobile phase: 0.225% aqueous formic acid/acetonitrile; flow rate: 30mL/min; gradient: 25% -100%,16 min) afforded compound 16.MS m/z (ESI): 597.1[ M+H ] ⁺.

Step 10: separation of Compound 16a and Compound 16b

Compound 16 (75 mg,0.13 mmol) was separated by supercritical fluid chromatography (column: CHIRALPAK AD i.d.,250 x 30mm,10 μm, mobile phase: carbon dioxide/isopropanol (0.1% ammonia) =60/40, flow rate: 150 mL/min) to give compound 16a (peak 1, retention time 0.870 min) and compound 16b (peak 2, retention time 1.484 min).

Compounds of formula (I) 16a：MS m/z(ESI):597.4[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.81(d,J＝4.0Hz,1H),7.76(s,1H),7.01(s,2H),6.81-6.80(m,2H),6.59(s,1H),5.53(t,J＝8.0Hz,1H),4.09(s,3H),3.93(s,3H),2.99-2.97(m,1H),2.75-2.73(m,1H),2.40-2.39(m,4H),2.18-2.16(m,1H),2.06-2.04(m,1H),1.82-1.64(m,11H),1.32(d,J＝4.0Hz,3H).

Compounds of formula (I) 16b：MS m/z(ESI):597.4[M+H]⁺.¹H NMR(400MHz,DMSO-d₆)δ:7.82(d,J＝4.0Hz,1H),7.76(s,1H),7.02(s,2H),6.81(d,J＝4.0Hz,2H),6.61(s,1H),5.54-5.51(m,1H),4.09(s,3H),3.93(s,3H),2.97-2.95(m,1H),2.77-2.75(m,1H),2.41-2.40(m,4H),2.15-2.13(m,1H),2.06-2.04(m,1H),1.85-1.66(m,11H),1.33(s,3H).

Example 18: preparation of Compound 18

Step1: synthesis of Compound 18-1

To a mixed solution of compound 17 (400 mg,0.86 mmol) in water (0.8 mL) and tetrahydrofuran (4 mL), potassium hydrogen peroxymonosulfate (0.06 mL,0.43 mmol) was added, and the reaction mixture was stirred at 10℃for 12 hours. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL. Times.3). The organic phase was washed with saturated brine (5 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and the crude product was purified by flash column chromatography on silica gel (eluent: dichloromethane/methanol) to give compound 18-1.MS m/z (ESI): 482.2[ M+H ] ⁺.

Step 2: synthesis of Compound 18-2

To a solution of compound 18-1 (360 mg,0.75 mmol) in N, N-dimethylformamide (4 mL) was added N, N-dimethylformamide dimethyl acetal (1.00 mL,7.50 mmol), and the reaction mixture was stirred at 10℃for 12 hours. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL. Times.3). The organic phase was washed with saturated brine (5 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and the crude product was purified by flash column chromatography on silica gel (eluent: dichloromethane/methanol) to give compound 18-2.MS m/z (ESI): 537.2[ M+H ] ⁺.

Step 3: synthesis of Compound 18-3

To a solution of compound 18-2 (100 mg,0.19 mmol) and compound A-1 (48.15 mg,0.37 mmol) in tetrahydrofuran (4 mL) was added sodium tert-butoxide (35.81 mg,0.37 mmol), and the reaction was stirred at 10℃for 1 hour. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL. Times.3). The organic phase was washed with saturated brine (5 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate to obtain compound 18-3.MS m/z (ESI): 602.5[ M+H ] ⁺.

Step 4: synthesis of Compound 18

A solution of compound 18-3 (100 mg,0.17 mmol) in methanol (2 mL, 4M) in hydrogen chloride was stirred at 50deg.C for 12 hours. The pH of the reaction solution was adjusted to 7 to 8 with ammonia water. The reaction mixture was concentrated and the crude product was purified by preparative high performance liquid chromatography (column: waters-SunFire-C18, 19X 250mm,10 μm; mobile phase: 0.1% aqueous formic acid-acetonitrile; gradient: acetonitrile 24% -95%,14 min) to give compound 18 (formate). MS m/z (ESI): 547.4[ M+H ] ⁺.

¹H NMR(400MHz,DMSO-d₆)δ:8.17(s,1H),7.70(d,J＝4.0Hz,1H),7.09(s,2H),6.87(d,J＝4.0Hz,1H),5.36-5.30(m,1H),4.98-4.91(m,1H),3.02-2.97(m,1H),2.76-2.71(m,1H),2.58-2.55(m,2H),2.44(s,3H),2.25-2.11(m,2H),2.00-1.95(m,1H),1.88-1.77(m,7H),1.71-1.64(m,2H),1.50-1.47(m,6H),1.30(d,J＝8.0Hz,3H).

Example 24: preparation of Compound 24

Step1: synthesis of Compound 24-2

To a solution of compound 18-2 (50 mg,0.09 mmol) and compound 24-1 (24.1 mg,0.18 mmol) in tetrahydrofuran (1 mL) was added sodium tert-butoxide (17.3 mg,0.18 mmol), and the reaction was stirred at 10℃for 1 hour. The reaction mixture was diluted with water (2 mL) and extracted with ethyl acetate (2 mL. Times.3). The organic phase was washed with saturated brine (2 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate to obtain compound 24-2.MS m/z (ESI): 602.4[ M+H ] ⁺.

Step 2: synthesis of Compound 24

A solution of compound 24-2 (50 mg,0.08 mmol) in methanol (1 mL, 4M) in hydrogen chloride was stirred at 50deg.C for 12 hours. The pH of the reaction solution was adjusted to 7 to 8 with ammonia water. The reaction mixture was concentrated and the crude product was purified by preparative high performance liquid chromatography (column: waters-Xbridge-C18, 19X 250mM,10 μm; mobile phase: 10mM ammonium bicarbonate aqueous solution-acetonitrile; gradient: 49% -95% acetonitrile, 14 min) to give compound 24.MS m/z (ESI): 547.4[ M+H ] ⁺.

¹H NMR(400MHz,DMSO-d₆)δ:7.71(d,J＝4.0Hz,1H),7.09(s,2H),6.88(d,J＝4.0Hz,1H),4.99-4.95(m,1H),4.28(s,2H),2.61-2.57(m,2H),2.35-2.30(m,2H),2.24-2.12(m,7H),2.01-1.97(m,1H),1.93-1.75(m,5H),1.47(d,J＝8.0Hz,6H),0.68(s,2H),0.43(s,2H).

Example 26: preparation of Compound 26

Step1: synthesis of Compound 26-2

To a solution of compound 18-2 (50 mg,0.09 mmol) and compound 26-1 (31.9 mg,0.19 mmol) in tetrahydrofuran (1 mL) was added sodium tert-butoxide (17.9 mg,0.19 mmol), and the reaction was stirred at 10℃for 1 hour. The reaction mixture was diluted with water (2 mL) and extracted with ethyl acetate (2 mL. Times.3). The organic phase was washed with saturated brine (2 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate to obtain compound 26-2.MS m/z (ESI): 644.4[ M+H ] ⁺.

Step 2: synthesis of Compound 26

A solution of compound 26-2 (50 mg,0.08 mmol) in methanol (1 mL, 4M) in hydrogen chloride was stirred at 50deg.C for 12 hours. The pH of the reaction solution was adjusted to 7 to 8 with ammonia water. The reaction mixture was concentrated and the crude product was purified by preparative high performance liquid chromatography (column: waters-SunFire-C18, 19X 250mm,10 μm; mobile phase: 0.1% aqueous formic acid-acetonitrile; gradient: acetonitrile 22% -95%,14 min) to give compound 26 (formate). MS m/z (ESI): 589.4[ M+H ] ⁺.

¹H NMR(400MHz,DMSO-d₆)δ:7.71(d,J＝4.0Hz,1H),7.09(s,2H),6.88(d,J＝4.0Hz,1H),5.00-4.93(m,1H),4.33(s,2H),3.52(t,J＝8.0Hz,4H),2.58-2.55(m,2H),2.45-2.36(s,4H),2.35-2.33(m,2H),2.18-2.12(m,1H),2.00-1.96(m,1H),1.89-1.84(m,5H),1.48(d,J＝8.0Hz,6H),0.69-0.66(m,2H),0.44-0.41(m,2H).

Example 30: preparation of Compound 30

Step1: synthesis of Compound 30-2

To a solution of compound 30-1 (5 g,31.62 mmol) in N, N-dimethylformamide (50 mL) was added sodium hydride (1.39 g,34.78mmol, purity 60%) at 0℃and stirred at 0℃for 1 hour. Methyl iodide (4.94 g,34.78 mmol) was then added and stirred at 25℃for 18 hours. The reaction mixture was diluted with water (100 mL), extracted with ethyl acetate (50 mL. Times.3), and the combined organic phases were washed with saturated brine (50 mL. Times.2). Drying and filtering with anhydrous sodium sulfate, concentrating, and purifying the crude product by using a rapid silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain a compound 30-2.

¹H NMR(400MHz,CDCl₃)δ:4.53(dd,J＝11.6,1.6Hz,1H),4.28-4.12(m,1H),3.81-3.61(m,4H),3.36(d,J＝11.6Hz,1H),2.89(ddd,J＝14.4,11.6,7.2Hz,1H),2.47(ddd,J＝14.4,3.2,

2.4Hz,1H),1.26(s,3H)。

Step 2: synthesis of Compound 30-3

To a solution of compound 30-2 (5.7 g,33.1 mmol) in toluene (50 mL) was added malononitrile (3.28 g,49.66 mmol), ammonium acetate (510 mg,6.62 mmol) and acetic acid (0.8 g,13.24 mmol). The reaction was stirred at 110℃for 18 hours. The reaction mixture was diluted with water (100 mL), extracted with ethyl acetate (50 mL. Times.3), and the combined organic phases were washed with saturated brine (50 mL. Times.2). Drying and filtering with anhydrous sodium sulfate, concentrating, and purifying the crude product by using a rapid silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain a compound 30-3.MS m/z (ESI): 221.1[ M+H ] ⁺.

Step 3: synthesis of Compound 30-4

To a solution of compound 30-3 (3.1 g,14.08 mmol) in N, N-dimethylformamide (50 mL) were added L-proline (320 mg,2.82 mmol) and sulfur (900 mg,28.15 mmol), and the reaction was stirred at 80℃for 18 hours. The reaction mixture was poured into water (100 mL). The aqueous layer was extracted with ethyl acetate (50 mL. Times.3), and the combined organic layers were washed with saturated brine (50 mL. Times.2). Drying and filtering with anhydrous sodium sulfate, concentrating, and purifying the crude product by using a rapid silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain a compound 30-4.MS m/z (ESI): 253.2[ M+H ] ⁺.

Step 4: synthesis of Compound 30-5

To a solution of compound 30-4 (1 g,3.96 mmol) in ethanol (10 mL) was added aqueous potassium hydroxide (4.95 mL, 4M), and the reaction was stirred at 80℃for 12 hours. The reaction solution was adjusted to pH 6-7 with hydrochloric acid and then concentrated. The solid was dissolved in methanol (10 mL), filtered and concentrated to give compound 30-5.MS m/z (ESI): 239.1[ M+H ] ⁺.

Step 5: synthesis of Compound 30-6

To a solution of compound 30-5 (493 mg,2.07 mmol) and 2- (7-azabenzotriazol) -N, N, N ', N' -tetramethylurea hexafluorophosphate (1468 mg,2.82 mmol) in N, N-dimethylformamide (10 mL) was added N, N-diisopropylethylamine (0.62 mL,3.76 mmol), followed by compound 17-5 (500 mg,1.88 mmol). The reaction solution was stirred at 10℃for 12 hours. The reaction mixture was diluted with water (10 mL), and extracted with ethyl acetate (10 mL. Times.3). The organic phase was washed with saturated brine (10 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by flash silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain the compound 30-6.MS m/z (ESI): 486.2[ M+H ] ⁺.

Step 6: synthesis of Compound 30-7

A solution of compound 30-6 (250 mg,0.51 mmol) and sodium carbonate (164 mg,1.54 mmol) in dimethyl sulfoxide (5 mL) was stirred at 100deg.C for 12 hours. The reaction mixture was diluted with water (5 mL) and extracted with ethyl acetate (5 mL. Times.3). The organic phase was washed with saturated brine (5 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate, and purifying the crude product by flash silica gel column chromatography (eluent: ethyl acetate/petroleum ether) to obtain the compound 30-7.MS m/z (ESI): 468.2[ M+H ] ⁺.

Step 7: synthesis of Compound 30-8

To a mixed solution of compound 30-7 (120 mg,0.26 mmol) in water (0.4 mL) and tetrahydrofuran (2 mL) was added potassium hydrogen peroxydisulfate (0.07 mL,0.13 mmol), and the reaction was stirred at 10℃for 12 hours. The reaction mixture was diluted with water (2 mL) and extracted with ethyl acetate (5 mL. Times.3). The organic phase was washed with saturated brine (2 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate to obtain compound 30-8.MS m/z (ESI) 484.2[ M+H ] ⁺.

Step 8: synthesis of Compound 30-9

To a solution of compound 30-8 (100 mg,0.21 mmol) in N, N-dimethylformamide (2 mL) was added N, N-dimethylformamide dimethyl acetal (0.28 mL,2.07 mmol), and the reaction mixture was stirred at 10℃for 12 hours. The reaction mixture was diluted with water (2 mL) and extracted with ethyl acetate (5 mL. Times.3). The organic phase was washed with saturated brine (2 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated and the crude product was purified by column chromatography on silica gel (eluent: dichloromethane/methanol) to give compound 30-9.MS m/z (ESI) 539.2[ M+H ] ⁺.

Step 9: synthesis of Compound 30-10

To a solution of compound 30-9 (90 mg,0.17 mmol) and compound A-1 (43.18 mg,0.33 mmol) in tetrahydrofuran (1 mL) was added sodium tert-butoxide (32.11 mg,0.33 mmol), and the reaction was stirred at 10℃for 1 hour. The reaction mixture was diluted with water (2 mL) and extracted with ethyl acetate (2 mL. Times.3). The organic phase was washed with saturated brine (2 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate to obtain compound 30-10.MS m/z (ESI) 604.4[ M+H ] ⁺.

Step 10: synthesis of Compound 30

A solution of compound 30-10 (100 mg,0.17 mmol) in methanol hydrochloride (2 mL, 4M) was stirred at 50deg.C for 12 hours. Concentrating the reaction solution, and purifying the crude product by preparative high performance liquid chromatography (chromatographic column: xtimate C18,150 ×25mm,5 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 25% -100%,17 min) to obtain compound 30 (hydrochloride). MS m/z (ESI): 549.1[ M+H ] ⁺.

¹H NMR(400MHz,DMSO-d₆)δ:10.17(s,1H),7.80(d,J＝4.0Hz,1H),7.32(s,2H),6.95(d,J＝4.0Hz,1H),5.43-5.36(m,1H),5.01-4.94(m,1H),4.62(s,2H),4.20-4.16(m,1H),3.91(d,J＝12.0Hz,1H),3.82-3.74(m,1H),3.59-3.53(m,1H),3.19-3.10(m,1H),3.01(dd,J＝8.0,4.0Hz,3H),2.28-2.23(m,1H),2.11-2.04(m,1H),1.97-1.86(m,2H),1.80(s,3H),1.50(d,J＝8.0Hz,6H),1.46(d,J＝8.0Hz,3H).

Example 32: preparation of Compound 32

Step1: synthesis of Compound 32-1

To a solution of compound 18-2 (50 mg,0.09 mmol) and compound 14-1 (29.66 mg,0.19 mmol) in tetrahydrofuran (1 mL) was added sodium tert-butoxide (17.91 mg,0.19 mmol), and the reaction was stirred at 10℃for 1 hour. The reaction mixture was diluted with water (2 mL) and extracted with ethyl acetate (2 mL. Times.3). The organic phase was washed with saturated brine (2 mL. Times.2), dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate to obtain compound 32-1.MS m/z (ESI): 632.4[ M+H ] ⁺.

Step 2: synthesis of Compound 32

A solution of compound 32-1 (50 mg,0.08 mmol) in methanol (1 mL, 4M) in hydrogen chloride was stirred at 50deg.C for 12 hours. The pH of the reaction solution was adjusted to 7 to 8 with ammonia water. Concentrating the reaction solution, and purifying the crude product by preparative high performance liquid chromatography (chromatographic column: xtimate C18,150 ×25mM,5 μm; mobile phase: 10mM ammonium bicarbonate/water-acetonitrile; gradient: acetonitrile 45% -100%,17 min) to obtain compound 32.MS m/z (ESI): 577.4[ M+H ] ⁺.

¹H NMR(400MHz,DMSO-d₆)δ:7.72(d,J＝3.6Hz,1H),7.12(s,2H),6.89(d,J＝3.6Hz,1H),5.40-5.19(m,1H),5.04-4.90(m,1H),4.22-4.00(m,2H),3.17-3.02(m,3H),2.89-2.79(m,1H),2.59-2.55(m,2H),2.20-1.96(m,5H),1.91-1.76(m,8H),1.47(d,J＝6.8Hz,6H).

Example 35: preparation of Compound 35

Step1: synthesis of Compound 35-2

Compound 35-1 (50.0 g,256 mmol) was dissolved in tetrahydrofuran (50 mL), a solution of magnesium dichloride (2, 6-tetramethylpiperidine) in lithium salt in tetrahydrofuran (282 mL, 1M) was stirred under nitrogen for 0.5 hours, a solution of 1, 2-dibromotetrachloroethane (125 g, 284 mmol) in tetrahydrofuran (10 mL) was added, and the mixture was reacted at 25℃for 0.5 hours. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride (200 mL), diluted with water (500 mL), extracted with ethyl acetate (500 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by flash column chromatography on silica gel (eluent: ethyl acetate/petroleum ether) to give compound 35-2.MS m/z (ESI): 274.8[ M+H ] ⁺.

Step 2: synthesis of Compound 35-3:

compound 35-2 (5.81 g,21.2 mmol) and triethylamine (6.43 g,63.6 mmol) were dissolved in ethanol (250 mL) and reacted at 25℃for 3 hours. Water (50 mL) was added to dilute, extraction was performed with ethyl acetate (50 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by flash column chromatography on silica gel (eluent: ethyl acetate/petroleum ether) to give compound 35-3.MS m/z (ESI): 341.9[ M+H ] ⁺.

Step 3: synthesis of Compound 35-4:

Compound 35-3 (5.10 g,14.9 mmol), cesium carbonate (14.6 g,44.9 mmol), 4, 5-bis (diphenylphosphorus) -9, 9-dimethylxanthene (3.46 g,5.99 mmol) was dissolved in dioxane (500 mL), and tris (dibenzylideneacetone) dipalladium (2.74 g,2.99 mmol) was added under nitrogen atmosphere and reacted at 100℃for 2 hours. Water (500 mL) was added to dilute, extraction was performed with ethyl acetate (500 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified using a flash column of silica gel (ethyl acetate/petroleum ether) to give compound 35-4.MS m/z (ESI): 259.8[ M+H ] ⁺.

Step 4: synthesis of Compound 35-5:

Compound 35-4 (1.70 g,6.54 mmol) and zinc cyanide (1.53 g,13.0 mmol) were dissolved in 1-methyl-2-pyrrolidone (30 mL), and tetrakis (triphenylphosphine) palladium (756 mg,0.65 mmol) was added under nitrogen atmosphere and reacted at 150℃for 40 minutes under microwaves. Water (50 mL) was added to dilute, extraction was performed with ethyl acetate (50 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified using a flash column of silica gel (eluent: ethyl acetate/petroleum ether) to give compound 35-5.MS m/z (ESI): 251.1[ M+H ] ⁺.

Step 5: synthesis of Compound 35-6:

compound 35-5 (740 mg,2.96 mmol) was dissolved in ethanol (8 mL), hydroxylamine hydrochloride (534 mg,7.69 mmol) was added, and the mixture was reacted at 85℃for 3 hours. Water (50 mL) was added for dilution, extraction with ethyl acetate (50 mL. Times.3) and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give compound 35-6.MS m/z (ESI):

284.0[M+H]⁺。

step 6: synthesis of Compound 35-7:

Compound 35-6 (700 mg,2.47 mmol) and compound A-2-P1 (583 mg,2.47 mmol) were dissolved in N, N-dimethylformamide (10 mL), N-diisopropylethylamine (957 mg,7.41 mmol) was added, the temperature was lowered to 0 ℃, and (benzotriazole-1-oxy) triphenylphosphine hexafluorophosphate (1.92 g,3.71 mmol) was added and then reacted at 25℃for 6 hours. Water (50 mL) was added to dilute, extraction was performed with ethyl acetate (50 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified using flash column chromatography on silica gel (methanol/dichloromethane) to give compound 35-7.MS m/z (ESI): 502.2[ M+H ] ⁺.

Step 7: synthesis of Compound 35-8:

Compound 35-7 (730 mg,1.47 mmol) was dissolved in N, N-dimethylformamide (20 mL) and reacted at 100℃for 24 hours. Water (50 mL) was added to dilute, extraction was performed with ethyl acetate (50 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified using a flash column of silica gel (ethyl acetate/petroleum ether) to give compound 35-8.MS m/z (ESI) 484.2[ M+H ] ⁺.

Step 8: synthesis of Compound 35-9:

Compound 35-8 (930 mg,1.92 mmol) was dissolved in methylene chloride (10 mL), and m-chloroperoxybenzoic acid (389 mg,1.92mmol, purity 85%) was added at 0℃to react the reaction mixture at 0℃for 1.5 hours. Water (50 mL) was added to dilute, extraction was performed with ethyl acetate (50 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified using a flash column of silica gel (eluent: methanol/dichloromethane) to give compound 35-9.MS m/z (ESI) 500.3[ M+H ] ⁺.

Step 9: synthesis of Compound 35:

(S) -1- ((S) -1-methylpyrrolidin-2-yl) ethan-1-ol (206 mg,1.60 mmol) was dissolved in tetrahydrofuran (2 mL), a 1.0M solution of lithium bis (trimethylsilyl) amide (1.6 mL,1.60 mmol) in tetrahydrofuran was added under a nitrogen atmosphere at 25℃and the reaction was stirred at 50℃for 1 hour. A solution of 35-9 (400 mg,0.80 mmol) of the compound in tetrahydrofuran (2 mL) was added dropwise to the reaction, and the reaction was carried out at 60℃for 3 hours. The reaction was quenched with saturated ammonium chloride solution (5 mL), extracted with ethyl acetate (50 mL. Times.3), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: boston Prime C18,150X 30mm,5mm; mobile phase: 0.225% formic acid in water-acetonitrile; gradient: 25% -45% acetonitrile, 16 min) to give compound 35 (formate). MS m/z (ESI): 565.4[ M+H ] ⁺.

¹H NMR(400MHz,DMSO-d₆)δ:8.20(s,1H),7.05(s,2H),5.11-5.03(m,1H),4.98-4.90(m,1H),4.20-4.13(m,2H),3.53-3.49(m,3H),3.01-2.94(m,1H),2.68-2.61(m,1H),2.55–2.53(m,2H),2.37(s,3H),2.23-2.15(m,1H),2.13-2.03(m,1H),1.96-1.82(m,2H),1.78(s,3H),1.76-1.70(m,2H),1.69-1.56(m,2H),1.23-1.11(m,9H).

According to any of the above embodiments of the invention, the following compounds are prepared from suitable starting materials:

biological Activity assay

Experimental example 1: SPR (Surface plasmon resonance) test

The purpose of the experiment is as follows: the affinity of the compounds of the invention for binding to KRAS G12V or KRAS WT was tested by the SPR method.

The test method comprises the following steps: SPR experiments were performed on the Biacore 8k system based on the method described by Tran et al (PNAS, 2020,117 (7): 3363-3364).

Firstly, the SA chip (GE HEALTHCARE) is preprocessed: three consecutive injections of 1mol/L NaCl mixed with 50mM NaOH in water, each lasting 1 minute; then equilibrated with HBST buffer (10mM HEPES,pH 7.5,150mM NaCl,5mM MgCl ₂, 0.5mM TCEP, 5. Mu.M GDP,0.05% T20). GDP-conjugated Biotin-labeled KRAS G12V protein (or other protein expressed in Beijing Kang Long, his-KRAS (G12V) -BTN, uniport: P01116-2 (T2-K169)) was injected, and the coupling amount was about 4000RU. Test compounds were diluted to a maximum concentration of 100. Mu.M in HBST buffer to a final DMSO concentration of 2% and diluted 1:1 in HBST buffer with 2% DMSO. Multicycle dose response: 30. Mu.L/min flow rate, binding time for each concentration of analyte of 90 seconds, dissociation time of 360 seconds; single cycle dose response: 30. Mu.L/min flow rate, binding time 200 seconds and dissociation time 2500 seconds. All experiments were performed at 25 ℃. The sensorgrams have been solvent corrected and data fitted using Biacore Insight evaluation software. The experimental results are shown in table 1.

TABLE 1 SPR affinity test results for partial Compounds of the invention with KRAS G12V

Test compounds	KD/nM
		Compound 1	39.8
Compound 1b	6.4
		Compound 3	20.3
Compound 7	19.2
		Compound 9	0.88
Compound 10	16.6
		Compound 11	0.25
Compound 11a	0.11
		Compound 11b	10.9
Compound 13	1.71
		Compound 14	11.2
Compound 15	4.63
		Compound 16	38.5
Compound 16a	24.6
		Compound 18	2.95

The above results indicate that the compounds of the present invention have a good affinity to the target mutein.

Experimental example 2: nucleotide exchange assay A

The purpose of the experiment is as follows: the effect of the compounds of the invention on KRAS G12V protein function was judged by examining the inhibition of SOS1 mediated KRAS protein nucleotide exchange process by the compounds of the invention.

The experimental steps are as follows:

Detection buffer (25 mM HEPES (pH 7.5)), 50mM NaCl aqueous solution, 10mM MgCl ₂ aqueous solution, 0.01% Brij-35) was prepared. Compound preparation was performed using Echo550 on a new 384 assay plate (Corning, cat#4512) at a maximum concentration of 100 μm, 3-fold dilution, 10 spots serially diluted, and repeated twice with a final DMSO concentration of 1%.

First, 5. Mu.L of KRAS G12V (expressed in Beijing Kang Long, his-KRAS (G12V) -BTN, uniport: P01116-2 (T2-K169)) to which MANT-GDP (Jenabioscience, accession number NU-205S) was bound was added to the assay plate at 1000rpm for 30S. The plates were allowed to equilibrate for 30 minutes at room temperature. After 30 minutes, data were recorded (raw data).

Second, 5. Mu.L of SOS1 (cytoskeleton, cat# CS-GE 02-XL) and GppNHp (Abcam, cat# ab 146659) mixed reagent were added to the compound wells and the negative control wells of the assay plate, and then 5. Mu. L GppNHp reagent was added to the positive control wells at 1000rpm and centrifuged for 30s. The plates were incubated for 180 min at room temperature.

Fluorescence signals (excitation 360nm, emission 448 nm) were read on SpectraMax (raw data).

Normalized data formula: rfu=raw data per well/raw data per well, four-parameter fitting of the data using GRAPHPAD PRISM was performed to calculate the IC ₅₀ values for the compounds, the results are shown in table 2,

TABLE 2 Nucleotide exchange assay test results for some of the compounds of the invention

Test compounds	IC50/μM
		Compound 1 (0027)	1.02
Compound 1a (P1)	4.52
		Compound 1b (P2)	7.21
Compound 3 (0033)	0.05
		Compound 4 (0034)	0.22
Compound 6 (0036)	0.08
		Compound 7 (0058)	0.05

Through detection, the compound has obvious inhibition effect on the functional process of the target mutant protein.

Experimental example 3: determination of cellular p-ERK inhibitory Activity

Test purpose: the method is used for detecting the inhibition effect of the compound on the cell p-ERK and evaluating the inhibition effect of the compound on the KRAS target.

Test purpose: the method is used for detecting the inhibition effect of the compound on the cell p-ERK and evaluating the inhibition effect of the compound on the KRAS target.

The test steps are as follows: human lung cancer cells NCI-H441 cells (number: HTB-174, ATCC) were cultured in complete medium, namely RPMI 1640 medium (Gibco, 11415) containing 10% fetal bovine serum (Corning, 35-081-CV) and 1% Penicillin-Streptomycin solution (Gibco, 15140122).

NCI-H441 cells were seeded in new white, bottom impermeable 384-well plates (Corning, 3570) at a density of 1000 cells/well using complete medium, 40 μl of cell suspension per well; placed at 37℃in a 5% CO2 cell incubator overnight.

Taking out 384-well cell culture plates, adding 40nL of compound to be tested into the cell culture plates by adopting an ultrasonic nano liter liquid treatment system Echo650, so that the final concentration of the compound is 1 mu M at most, diluting 3 times, continuously diluting 10 points, repeating twice, and finally obtaining the DMSO concentration of 0.1%. Placed in a 37℃5% CO2 cell incubator for 3 hours.

The intracellular p-ERK content assay was performed using the Advanced Phospho-ERK 1/2 (Thr 202/Tyr 204) kit (Cisbio, 64 AERPEG). Preparation of lysia buffer (1 x, kit components): 1 volume of lysis buffer (4 x means four times the final concentration, i.e. four times the original concentration) is mixed with 3 volumes of ultrapure water for standby; preparation Premixed antibody solution: mixing Eu Cryptate antibody or d2 anti body with 19 times of detection buffer for use.

Taking out 384-well plates, and discarding the supernatant; add 16. Mu.L of lysis buffer (1X), incubate with shaking at room temperature for at least 30 minutes; centrifuge at 3000rpm for 2 minutes. Adding 4 mu L Premixed antibody solution into each hole, and uniformly mixing for 30 seconds; centrifuge at 1000rpm for 5 minutes. Incubate at room temperature for 2.5 hours. Fluorescence signals were read on Envision HTS Multilabel Reader (excitation 337nm, emission 620nm/665 nm).

Wells containing 1 μm Staurosporine (test ginseng, MCE, cat No. HY-15141) were defined as 100% inhibited (PC), wells containing 0.1% dmso final concentration as 0% inhibited (NC); inhibition% = (avg.nc-rfu.cpd)/(avg.nc-avg.pc) x100% (rfu.cpd refers to the relative fluorescence value for each well of the compound group, i.e. emission 620nm/665 nm). The data were subjected to concentration-inhibition nonlinear regression analysis of the test compounds in GRAPHPAD PRISM to obtain IC ₅₀ values for the test compounds. The test results are shown in Table 3. The test result shows that the compound has obvious inhibition effect on the p-ERK function of target cells.

Experimental example 4: determination of cell antiproliferative Activity

Test purpose: is used for detecting the inhibition of the proliferation of human lung cancer cells NCL-H441 by the compound.

The experimental steps are as follows: human lung cancer cells NCI-H441 (accession No. HTB-174, ATCC) were cultured in complete medium, namely RPMI 1640 medium (Gibco, 22400089) containing 10% fetal bovine serum (Corning, 35-081-CV) and 1% Penicillin-Streptomycin solution (Gibco, 15140122).

Cells were seeded into new 384-well cell culture plates (Greiner, 781091) at 1000 per well using complete medium, 60 μl of cell suspension per well; placed at 37℃in a 5% CO2 cell incubator overnight. The compound to be tested was diluted with complete medium and 20 μl of compound was added per well to give a final concentration of up to 10 μm, diluted 3-fold, serially diluted 10 spots, repeated twice and final DMSO concentration of 0.25%. Centrifuge at 1000rpm at room temperature for 1min. Placed in a 37℃5% CO2 cell incubator for 5 days.

Taking out 384-well cell culture plates, and discarding 40 mu L of supernatant; subsequently, 20. Mu.L of a luminescent cell activity detection reagent (Promega, G7573) was added, and after shaking for 2 minutes in the dark, centrifugation was carried out at 1000rpm for 1 minute; placing for 10 minutes at room temperature in a dark place; the luminescence signal value (lumi.cpd) is read at Envision HTS Multilabel Reader.

Wells containing 1 μm Staurosporine (test ginseng, MCE, cat No. HY-15141) were defined as 100% inhibited (PC), wells containing 0.25% dmso final concentration as 0% inhibited (NC); inhibition% = (avg.nc-lumi.cpd)/(avg.nc-avg.pc) x100%. The data were subjected to concentration-inhibition nonlinear regression analysis of the test compounds in GRAPHPAD PRISM to obtain IC ₅₀ values for the compounds, and the test results are shown in table 3. The test result shows that the compound has obvious inhibition effect on the proliferation process of target cells.

TABLE 3 determination of inhibitory Activity and cell anti-proliferative Activity of Compounds of the invention

Claims (15)

1. A compound of formula I or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein ring B is a 5-6 membered heteroaryl group; Ring A and Ring C are fused together to form a 7-12-membered heterobicyclic group, wherein Ring A is a 5-6-membered heteroaromatic ring, and Ring C is selected from a benzene ring, a 5-6-membered heteroaromatic ring, and a 5-6-membered heterocyclic ring; Ra is independently selected from halogen, C _1-4 alkyl, C _1-4 alkyl-S(O) ₂ -, 3-10 membered heterocyclyl, 3-10 membered heterocyclyl-(CR ¹ R ² )q ₁ -X ¹ -, N(R ³ )(R ⁴ )-(CR ⁵ R ⁶ )q ₂ -X ² -; wherein the heterocyclyl is optionally substituted by one or more of the following groups: halogen, hydroxyl, C _1-4 alkyl, C _1-4 alkoxy-C _1-4 alkyl-, -N(R ⁷ )(R ⁸ ), optionally substituted 3-6 membered heterocyclyl; X ¹ and X ² are independently selected from a chemical bond and O; q ₁ and q ₂ are independently selected from 0, 1, 2, and 3; R ¹ and R ² are independently selected from H and C _1-4 alkyl; R ³ and R ⁴ are independently selected from H, C _1-4 alkyl, or R ³ , R ⁴ and the N atom to which they are connected form a 4-6 membered heterocyclic ring; R ⁵ and R ⁶ are independently selected from H, C _1-4 alkyl, or R ⁵ and R ⁶ on the same C atom and the C atom to which they are attached form an optionally substituted 3-6 membered cycloalkyl; R ⁷ and R ⁸ are independently selected from H and C _1-4 alkyl; Rc is independently selected from H, C _1-4 alkyl, CN-C _1-4 alkyl, -(CR ⁹ R ¹⁰ )s-Cy, N(R ¹¹ )(R ¹² )-C _1-4 alkyl-; Cy is selected from the following groups optionally substituted with 1-3 Rx: 5-10 membered heteroaryl, phenyl, 3-6 membered heterocyclyl, 3-6 membered cycloalkyl, 5-6 membered cycloalkenyl; Rx is selected from oxo, hydroxy, halogen, C _1-4 alkyl, -N(R ¹³ )(R ¹⁴ ); s is selected from 0, 1, 2, 3; R ⁹ and R ¹⁰ are independently selected from H, C _1-4 alkyl, halogen, halogenated C _1-4 alkyl, C _3-6 cycloalkyl, or R ⁹ and R ¹⁰ on the same carbon atom together with the C atom to which they are attached form a 3-6 membered cycloalkyl; R ¹¹ , R ¹² , R ¹³ , and R ¹⁴ are independently selected from H, C _1-4 alkyl, deuterated C _1-4 alkyl, and halogenated C _1-4 alkyl; m and n are independently selected from 0, 1, 2, and 3. 2. The compound of formula I according to claim 1 or its pharmaceutically acceptable salt, or its stereoisomer, characterized in that ring B is selected from the following groups: * indicates that the position is connected to ring A. 3. The compound of formula (I) according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein ring A is selected from a pyridine ring or a pyrimidine ring. 4. The compound of formula (I) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein ring C is selected from a pyrrole ring, a benzene ring, a pyridine ring, a pyridazine ring, or an imidazole ring. 5. The compound of formula I according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that Ra is independently selected from the following groups: Cl, F, -S(O) ₂ CH ₃ , 6. The compound of formula I according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Rc is independently selected from H, -CH ₃ , -CH(CH ₃ ) ₂ , CNCH ₂ CH ₂ -, cyclopentyl, 7. The compound of formula I according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that the structural unit Selected from wherein Ra, Rc, m, and n are as defined in any one of claims 1 to 6. 8. The compound of formula I according to claim 7 or its pharmaceutically acceptable salt, or its stereoisomer, characterized in that the structural unit Selected from wherein Ra and Rc are as defined in claim 7. 9. The compound of formula I according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that the compound has the following structure: Wherein, Ra and Rc are as defined in any one of claims 1 to 8; Z ¹ and Z ² are independently selected from CH and N, and at least one is selected from N; Alternatively, in a technical solution of Formula Id or Formula Id-1 of the present invention, Z1 and Z2 are both N; Alternatively, in a technical solution of Formula Id or Formula Id-1 of the present invention, Ra is a 3-10 membered heterocyclic group -CR ¹ R ² -O-; said R ₁ and R ₂ are independently selected from H, methyl, ethyl, isopropyl, trifluoromethyl, trichloromethyl; said 3-10 membered heterocyclic group is optionally substituted by one or more of the following groups: halogen, hydroxyl, C _1-4 alkyl, C _1-4 alkoxy-C _1-4 alkyl-, -NH ₂ , an optionally substituted 3-6 membered heterocyclic group; preferably, Ra is a 5 membered heterocyclic group -CH(R ₂ )-O-; said 5 membered heterocyclic ring may be optionally substituted by halogen, hydroxyl, C _1-4 alkyl; Alternatively, in a technical solution of Formula Id or Formula Id-1 of the present invention, Ra is selected from Alternatively, in a technical solution of Formula Id or Formula Id-1 of the present invention, Ra is Alternatively, in one embodiment of Formula Id or Formula Id-1 of the present invention, Rc is independently selected from H, C _1-4 alkyl, -(CR ⁹ R ¹⁰ )-Cy; Cy is selected from the following groups optionally substituted by 1-3 Rx: 5-10 membered heteroaryl, phenyl; Rx is selected from oxo, hydroxyl, halogen, C _1-4 alkyl, -NH ₂ ; R ⁹ and R ¹⁰ are independently selected from H, C _1-4 alkyl, halogen, halogenated C _1-4 alkyl, C _3-6 cycloalkyl; Alternatively, in a technical solution of Formula Id or Formula Id-1 of the present invention, Rc is a pyridyl group substituted with one or more Rx; Alternatively, in a technical solution of Formula Id or Formula Id-1 of the present invention, Rc is a pyridyl group substituted with one or more Rx; Alternatively, in one technical solution of Formula Id or Formula Id-1 of the present invention, Rc is selected from methyl; 1-(2-aminopyridin-3-yl)-ethyl; Alternatively, in a technical solution of Formula Id or Formula Id-1 of the present invention, it has a structure as shown in the following Formula Id-2 or Formula Id-3. 10. The compound of formula I according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that it has the following structure: Wherein, R ₁₀ is selected from H, C _1-4 alkyl, halogen, halogenated C _1-4 alkyl, C _3-6 cycloalkyl; Rx is selected from hydroxyl, halogen, C _1-4 alkyl, -NH ₂ ; Alternatively, in a technical solution of formula Id-2 or formula Id-3 of the present invention, R ₁₀ is selected from H, methyl, trifluoromethyl; Alternatively, in a technical solution of formula Id-2 or formula Id-3 of the present invention, Rx is selected from methyl, F, NH ₂ ; Alternatively, in one technical solution of Formula Id-2 or Formula Id-3 of the present invention, Rx is selected from NH ₂ ; Alternatively, in one technical solution of Formula Id-2 or Formula Id-3 of the present invention, R ₁₀ is selected from H or methyl, and Rx is NH ₂ . 11. The compound of formula I according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that it is selected from the following compounds or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: In one technical solution of the present invention, the compound of the present invention is preferably the following compound or a pharmaceutically acceptable salt thereof: In one technical solution of the present invention, the compound of the present invention is preferably the following compound or a pharmaceutically acceptable salt thereof: 12. A pharmaceutical composition comprising the compound of formula I according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, and any pharmaceutically acceptable carrier. 13. Use of the compound of formula I according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a drug for treating a disease associated with KRAS G12V gene mutation. 14. Use of the pharmaceutical composition according to claim 12 in the preparation of a drug for treating a disease associated with KRAS G12V gene mutation. 15. A method for treating a disease associated with KRAS G12V gene mutation, the method comprising administering an effective amount of the compound of formula I according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or the pharmaceutical composition according to claim 12 to a patient in need thereof.