WO2022152316A1

WO2022152316A1 - Biphenyl derivative and use thereof

Info

Publication number: WO2022152316A1
Application number: PCT/CN2022/072583
Authority: WO
Inventors: 吴凌云; 王才林; 徐雄彬; 童海骏; 陈曙辉
Original assignee: 南京明德新药研发有限公司
Priority date: 2021-01-18
Filing date: 2022-01-18
Publication date: 2022-07-21
Also published as: CN116710460A

Abstract

A biphenyl derivative, a preparation method therefor and the use thereof in the preparation of a medicine for treating related diseases. In particular, the present application relates to a compound represented by formula (I) and a pharmaceutically acceptable salt thereof.

Description

Biphenyl derivatives and their applications

This application claims the following priority:

CN202110066211.5, application date January 18, 2021.

technical field

The present invention relates to a biphenyl derivative, its preparation method and its application in preparation and treatment of related diseases, in particular to a compound represented by formula (I) and a pharmaceutically acceptable salt thereof.

Background technique

Programmed cell death 1 (PD-1), also known as CD279, is an important immunosuppressive molecule in the CD28/CTLA-4 receptor family. It is a membrane protein containing 268 amino acid residues and is widely expressed in T. On the surface of various immune cells such as cells, macrophages, and B cells, its ligands are PD-L1 and PD-L2. PD-L1 is a protein encoded by the CD274 gene and mainly expressed on the surface of tumor cells, dendritic cells and macrophages. After the combination of PD-1 and PD-L1, the PD-1/PD-L1 signaling pathway is activated, which in turn inhibits the activation of T cells, causing T cell inactivation and contributing to the immune escape of tumor cells. Another ligand of PD-1, PD-L2, is mainly expressed on the surface of dendritic cells, macrophages and B cells and is associated with inflammation and autoimmune diseases.

PD-1 negatively regulates immune responses by binding to its ligand PD-L1 and dephosphorylating multiple key molecules in the TCR signaling pathway. In healthy organisms, the activation of PD-1/PD-L1 signaling pathway can avoid the surrounding tissue damage caused by excessive immune response, thereby reducing the occurrence of autoimmune diseases. However, under the induction of the tumor microenvironment, the expressions of PD-1 and PD-L1 are abnormally increased, and tumor cells can successfully escape the recognition and detection of the immune system by the binding of these PD-L1 molecules to PD-1 on T cells. attack. PD-(L)1 mAb can block this "tumor immune escape mechanism" and restore the patient's own immune system's anti-cancer function.

Currently, the marketed drugs targeting the PD-1/PD-L1 pathway are monoclonal antibodies (mAbs). In 2014, the U.S. Food and Drug Administration (FDA) approved the first two monoclonal antibodies (Pembrolizumab and Nivolumab) to target the PD-1 molecule to the market. Over the next few years, three additional mAbs targeting the PD-L1 molecule (Atezolizumab, Durvalumab, and Avelumab) appeared on the market. All of the above are biological macromolecules, which have their own significant defects, such as being easily decomposed by proteases, poor in vivo stability, and need to be administered by injection; product quality is not easy to control, and production technology requirements are high; large-scale preparation and purification are difficult , high production cost and easy to produce immunogenicity. Small molecule PD-1/PD-L1 inhibitors have attracted more and more attention. Incyte's PD-L1 small molecule inhibitor INCB86550 (WO2018119263, WO2019191707) and Gilead's PD-L1 small molecule inhibitor GS-4224 (US20180305315, WO2019160882) ) has entered the clinical phase 2, and the small molecule PD-1/PD-L1 inhibitor of BMS benzyl phenyl ether (WO2015034820, WO2015160641) is in the preclinical research stage. Because small-molecule drugs can cross the cell membrane and act on intracellular targets, they have the advantages of convenient storage and transportation, low production cost, no immunogenicity, and usually oral administration. Therefore, research and development of small molecule blockers of PD-1/PD-L1 has broad application prospects.

SUMMARY OF THE INVENTION

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

in,

R ₁ and R ₂ are each independently selected from H, F, Cl, Br, I, CN and C _1-3 alkyl optionally substituted with ₁ , 2 or 3 halogens;

R ₃ is selected from H, CN, C _1-3 alkyl, C _1-3 alkoxy and C _1-3 alkylamino, the C _1-3 alkyl, C _1-3 alkoxy and C _{1- 3} alkylamino groups are each independently optionally substituted with 1, 2 or 3 halogens;

R ₄ and R ₅ are each independently selected from H, C _1-6 alkyl, C _1-6 alkylamino, C _3-6 cycloalkyl and -C _1-3 alkyl-(3-6 membered heterocycloalkane base), the C _1-6 _{alkyl, C 1-6} _alkylamino , C _3-6 cycloalkyl and -C _1-3 alkyl-(3-6 membered heterocycloalkyl) are independently optional is substituted with 1, 2 or 3 R _a ; alternatively, R ₄ and R ₅ are joined to form a 3-8 membered heterocycloalkyl optionally substituted by 1, 2 or 3 R _b replace;

R ₆ is selected from C _1-3 alkyl optionally substituted with 1, 2 or 3 R _c ;

L is selected from -C _1-6 alkyl- optionally substituted with 1, 2 or 3 R _d ;

X is selected from CH and N;

Y is selected from CH and N;

Z ₁ is selected from a single bond and CH ₂ ;

Z ₂ is selected from CH and N;

_Ra is selected from F, Cl, Br, I, OH, =O, NH ₂ , C _1-3 alkylamino, C _1-3 alkoxy and C _1-3 alkyl, the C _1-3 alkylamino , C _1-3 alkoxy and C _1-3 alkyl are each independently optionally substituted with 1, 2 or 3 R;

R _b is selected from CN, F, Cl, Br, I, OH, -C(=O)NH ₂ , C _1-3 alkylamino, C _1-3 alkoxy and C _1-3 alkyl;

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

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

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

The 3-6 membered heterocycloalkyl group and the 3-8 membered heterocycloalkyl group each independently contain 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from N, O, S and NH.

In some aspects of the present invention, the above Z ₂ is selected from CH, and other variables are as defined in the present invention.

In some aspects of the present invention, the above R ₁ and R ₂ are independently selected from H, F, Cl, Br, I, CN, CF ₃ and CH ₃ , and other variables are as defined in the present invention.

In some aspects of the invention, the above R ₃ is selected from H, CN, CH ₃ , -OCH ₃ and

The CH ₃ , -OCH ₃ and

Each independently is optionally substituted with 1, 2 or 3 halogens, other variables are as defined herein.

In some aspects of the present invention, the above R ₃ is selected from H, CN, CH ₃ , -OCH ₃ ,

Other variables are as defined in the present invention.

In some aspects of the present invention, the above R ₄ and R ₅ are each independently selected from H, C _1-4 alkyl, C _1-3 alkylamino, cyclopropyl and

The C _1-4 alkyl, C _1-3 alkylamino, cyclopropyl and

Each independently is optionally substituted with 1, 2 or 3 R _a , other variables are as defined in the present invention.

In some embodiments of the present invention, the above R _a is selected from F, Cl, Br, I, OH, =O, NH ₂ , -NHCH ₃ , -OCH ₃ , -CH ₂ OH and CH ₃ , other variables are as in the present invention defined.

In some aspects of the present invention, the above R ₄ and R ₅ are independently selected from H,

Other variables are as defined in the present invention.

In some aspects of the present invention, above-mentioned R ₄ is selected from H,

Other variables are as defined in the present invention.

In some aspects of the present invention, above-mentioned R ₅ is selected from H,

Other variables are as defined in the present invention.

In some embodiments of the invention, R4 and R5 described above are linked to form pyrrolidinyl, ₈ _- azabicyclo[3.2.1]octyl, azetidinyl and 2-azaspiro[3.3]heptyl alkyl, the pyrrolidinyl, 8-azabicyclo[3.2.1]octyl, azetidinyl, and 2-azaspiro[3.3]heptyl, respectively, independently optionally replaced by 1, 2 or 3 R _b substitutions, other variables are as defined in the present invention.

In some aspects of the present invention, the above R _b is selected from CN, OH, -C(=O) _NH2 , _-OCH3 and _CH3 , and other variables are as defined herein.

In some aspects of the invention, the above-mentioned R ₄ and R ₅ are linked to form

Other variables are as defined in the present invention.

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

In some aspects of the invention, the above L is selected from

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned structural units

selected from

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned structural units

selected from

Other variables are as defined in the present invention.

In some aspects of the invention, the above-mentioned compounds are selected from

Wherein, R ₁ , R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , Z ₁ are as defined in the present invention, R ₇ and R ₈ are selected from H, F, Cl, Br, I, CN, OH, = O and _NH2 .

wherein R ₁ , R ₂ , R ₃ , R ₆ , Z ₁ , Z ₂ , X and Y are as defined in the present invention,

Ring A is selected from 3-8 membered heterocycloalkyl,

R ₉ and R ₁₀ are each independently selected from H, CN, F, Cl, Br, I, OH, -C(=O)NH ₂ , C _1-3 alkylamino, C _1-3 alkoxy and C _{1 -3} alkyl.

The present invention also provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from

The present invention also provides the use of the above compounds or their pharmaceutically acceptable salts in the preparation of PD-1/PD-L1 inhibitors.

In some aspects of the present invention, the above PD-1/PD-L1 inhibitor is an anti-tumor drug.

technical effect

The compound of the present invention has a good inhibitory effect on the excessive activation of the PD-1/PD-L1 signaling pathway, thereby obtaining excellent tumor growth inhibitory activity.

Definition and Explanation

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered indeterminate or unclear without specific definitions, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commercial product or its active ingredient.

As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue , without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts including, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, and methanesulfonic acids; also include salts of amino acids such as arginine, etc. , and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and thus can be converted into either base or acid addition salts.

The pharmaceutically acceptable salts of the present invention can be synthesized from the acid or base containing parent compound by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.

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

Unless otherwise indicated, the terms "enantiomers" or "optical isomers" refer to stereoisomers that are mirror images of each other.

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

Unless otherwise indicated, the term "diastereomer" refers to a stereoisomer in which the molecule has two or more chiral centers and the molecules are in a non-mirror-image relationship.

Unless otherwise specified, "(+)" means dextrorotatory, "(-)" means levorotatory, and "(±)" means racemic.

Use solid wedge keys unless otherwise specified

and wedge-dotted keys

Indicate the absolute configuration of a stereocenter, using a straight solid key

and straight dashed keys

Indicate the relative configuration of the stereocenter, with a wavy line

Represents a solid wedge key

or wedge-dotted key

or with wavy lines

Represents a straight solid key

and straight dashed keys

Unless otherwise specified, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium and are rapidly interconverted at room temperature. A chemical equilibrium of tautomers can be achieved if tautomers are possible (eg, in solution). For example, proton tautomers (also called prototropic tautomers) include interconversions by migration of protons, such as keto-enol isomerization and imine-ene Amine isomerization. Valence tautomers include interconversions by recombination of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between two tautomers of pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

Unless otherwise indicated, the terms "enriched in one isomer", "enriched in isomers", "enriched in one enantiomer" or "enriched in one enantiomer" refer to one of the isomers or pairs The enantiomer content is less than 100%, and the isomer or enantiomer content is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or Greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

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

Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, followed by conventional methods known in the art The diastereoisomers were resolved and the pure enantiomers recovered. In addition, separation of enantiomers and diastereomers is usually accomplished by the use of chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (eg, from amines to amino groups) formate).

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, compounds can be labeled with radioisotopes, such as tritium ( ³ H), iodine-125 ( ¹²⁵ I) or C-14 ( ¹⁴ C). For another example, deuterated drugs can be formed by replacing hydrogen with deuterium, and the bonds formed by deuterium and carbon are stronger than those formed by ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All transformations of the isotopic composition of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

The terms "optional" or "optionally" mean that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. .

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

When any variable (eg, R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may optionally be substituted with up to two Rs, with independent options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

When the number of a linking group is 0, such as -(CRR) ₀ -, it means that the linking group is a single bond.

When one of the variables is selected from a single bond, it means that the two groups connected to it are directly connected, for example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.

When a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A. When the listed substituents do not indicate through which atom it is attached to the substituted group, such substituents may be bonded through any of its atoms, for example, pyridyl as a substituent may be through any one of the pyridine ring The carbon atom is attached to the substituted group.

When the listed linking group does not indicate its direction of attachment, the direction of attachment is arbitrary, for example,

The linking group L in the middle is -MW-, at this time -MW- can connect ring A and ring B in the same direction as the reading order from left to right.

It is also possible to connect ring A and ring B in the opposite direction to the reading order from left to right.

Combinations of the linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, when a group has one or more attachable sites, any one or more sites in the group can be linked to other groups by chemical bonds. When the connection method of the chemical bond is not located, and there is an H atom at the linkable site, when the chemical bond is connected, the number of H atoms at the site will be correspondingly reduced with the number of chemical bonds connected to the corresponding valence. the group. The chemical bond connecting the site to other groups can be represented by straight solid line bonds

straight dotted key

or wavy lines

express. For example, a straight solid bond in -OCH ₃ indicates that it is connected to other groups through the oxygen atom in this group;

The straight dashed bond in the group indicates that it is connected to other groups through the two ends of the nitrogen atom in the group;

The wavy line in the phenyl group indicates that it is connected to other groups through the 1 and 2 carbon atoms in the phenyl group;

Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least

These 4 connection methods, even if the H atom is drawn on -N-, but

still includes

The group in this connection method is only that when one chemical bond is connected, the H at the site will be correspondingly reduced by one to become the corresponding monovalent piperidinyl group.

Unless otherwise specified, the number of atoms in a ring is generally defined as the number of ring members, eg, "5-7 membered ring" refers to a "ring" of 5-7 atoms arranged around it.

Unless otherwise specified, the term "C _1-6 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. The C _1-6 alkyl includes C _1-5 , C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ and C ₅ alkyl and the like; it can be Is monovalent (eg methyl), divalent (eg methylene) or polyvalent (eg methine). Examples of C _1-6 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.

Unless otherwise specified, the term "C _1-4 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms. The C _1-4 alkyl includes C _1-2 , C _1-3 and C _2-3 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent ( such as methine). Examples of C _1-4 alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl , s-butyl and t-butyl) and so on.

Unless otherwise specified, the term "C _1-3 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, etc.; it can be monovalent (eg methyl), divalent (eg methylene) or multivalent (eg methine) . Examples of _C1-3 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

Unless otherwise specified, the term "C1-6alkoxy" refers to those _alkyl groups containing 1 to 6 carbon atoms attached to the remainder of the molecule through an oxygen atom. The C _1-6 alkoxy groups include C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ , C ₅ , C ₄ and C ₃ alkoxy groups, etc. . Examples of C _1-6 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n _- propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy) oxy, s-butoxy and t-butoxy), pentyloxy (including n-pentyloxy, isopentyloxy and neopentyloxy), hexyloxy and the like.

Unless otherwise specified, the term " _C1-3alkoxy " refers to those alkyl groups containing 1 to 3 carbon atoms attached to the remainder of the molecule through an oxygen atom. The C _1-3 alkoxy group includes C _1-2 , C _2-3 , C ₃ and C ₂ alkoxy and the like. Examples of C _1-3 alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

Unless otherwise specified, the term "C _1-6 alkylamino" refers to those alkyl groups containing 1 to 6 carbon atoms attached to the remainder of the molecule through an amino group. The C _1-6 alkylamino includes C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ , C ₅ , C ₄ , C ₃ and C ₂ alkylamino Wait. Examples of C _1-6 alkylamino include, but are not limited to, -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -N(CH ₂ CH ₃ )( _CH2CH3 ₎ , _-NHCH2CH2CH3 , _-NHCH2 ₍ _CH3 ₎ ₂ _, _{-NHCH2CH2CH2CH3} _, _etc.

Unless otherwise specified, the term "C _1-3 alkylamino" refers to those alkyl groups containing 1 to 3 carbon atoms attached to the remainder of the molecule through an amino group. The C _1-3 alkylamino groups include C _1-2 , C ₃ and C ₂ alkylamino groups and the like. Examples of C _1-3 alkylamino include, but are not limited to, -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -NHCH ₂ CH ₂ CH ₃ , - NHCH ₂ (CH ₃ ) ₂ and the like.

Unless otherwise specified, the term "halogen" or "halogen" by itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.

Unless otherwise specified, "C _3-6 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which are monocyclic and bicyclic ring systems, said C _3-6 cycloalkyl including C _3-5 , C _4-5 and C _5-6 cycloalkyl and the like; it may be monovalent, divalent or polyvalent. Examples of _C3-6 cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

Unless otherwise specified, the term "3- to 8-membered heterocycloalkyl" by itself or in combination with other terms denotes a saturated cyclic group consisting of 3 to 8 ring atoms, respectively, of which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, where the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (ie, NO and S(O) _p , p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spiro, paracyclic and bridged rings. Furthermore, with respect to the "3-8 membered heterocycloalkyl", a heteroatom may occupy the position of attachment of the heterocycloalkyl to the rest of the molecule. The 3-8 membered heterocycloalkyl includes 3-6 membered, 3-5 membered, 4-6 membered, 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyl and the like. Examples of 3-8 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl ( Including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2- piperidinyl and 3-piperidyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), Dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperyl pyridyl or dioxepanyl, etc.

Unless otherwise specified, the term "3-6 membered heterocycloalkyl" by itself or in combination with other terms denotes a saturated cyclic group consisting of 3 to 6 ring atoms, respectively, of which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the carbon, nitrogen, and sulfur heteroatoms are optionally oxidized (i.e., C(=O), NO and S(O)p, where p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein bicyclic ring systems include spiro, paracyclic and bridged rings. Furthermore, with respect to the "3-6 membered heterocycloalkyl", a heteroatom may occupy the position of attachment of the heterocycloalkyl to the remainder of the molecule. The 3-6 membered heterocycloalkyl includes 4-6 membered, 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyl and the like. Examples of 3-6 membered heterocycloalkyl include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothienyl ( Including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2- piperidinyl and 3-piperidyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), Dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl or homopiperazinyl pyridyl, etc.

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.

The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction method (SXRD), the cultured single crystal is collected by Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is:

After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.

The solvent used in the present invention is commercially available.

Unless otherwise specified, the proportions of reagents used in silica gel column chromatography, silica gel column chromatography and silica gel thin-layer chromatography plates in the present invention are all volume ratios.

The following abbreviations are used in the present invention: HATU stands for O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate; DMSO stands for di- Methyl sulfoxide; _CD3OD for deuterated methanol; _CDCl3 for deuterated chloroform; TBSO for tert-butyldimethylsilyloxy.

Compounds are named according to conventional nomenclature in the art or are used

Software naming, commercially available compounds use supplier catalog names.

Detailed ways

The present invention will be described in detail by the following examples, but it does not mean any unfavorable limitation of the present invention. The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention.

Intermediate A

first step

Compound A-1 (10 g, 48.43 mmol) and compound A-2 (15.99 g, 62.96 mmol) were dissolved in dioxane (100 mL), and 1,1′-bis( Diphenylphosphorus) ferrocene palladium chloride (3.54 g, 4.84 mmol) and potassium acetate (11.88 g, 121.08 mmol), the reaction mixture was replaced with gas and stirred at 80 °C for 15 hours under nitrogen protection. After the reaction was completed, the The reaction mixture was filtered and concentrated to obtain the crude product Intermediate A. MS-ESI calculated [M+H] ⁺ 254, found 254.

Intermediate B

first step

Compound B-1 (20 g, 144.80 mmol) and ethyl pyruvate (84.07 g, 723.99 mmol) were mixed and stirred at 20°C for 15 minutes. Phosphorus oxychloride (222.02 g, 1.45 mol) was then added and stirred at 100°C for 1 hour. After the reaction was completed, the reaction solution was poured into ice water (1 L), the pH was adjusted to 7 with sodium carbonate, extracted with ethyl acetate (150 mL×2), the organic phase was washed with saturated brine (100 mL×2), and the organic layer was It was dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=15:1 to 3:1) to obtain compound B-2. MS-ESI calculated [M+H] ⁺ 236.8 found 236.8.

second step

Compound B-2 (16.8 g, 70.99 mmol) and sodium cyanoborohydride (13.387 g, 212.97 mmol) were dissolved in acetic acid (100 mL), and the reaction solution was reacted at 20°C for 1 hour under nitrogen protection. After the reaction was completed, the reaction solution was poured into ice water, the pH was adjusted to 7 with sodium carbonate, extracted with ethyl acetate (200 mL×2), the organic phase was washed with saturated brine (100 mL×2), and dried over anhydrous sodium sulfate. , filtered and concentrated to give compound B-3 which was used directly in the next step without further purification. MS-ESI calculated [M+H] ⁺ 240.8, found 240.8.

third step

Compound B-3 (23 g, 95.56 mmol) was dissolved in dichloromethane (200 mL), di-tert-butyl dicarbonate (31.28 g, 143.34 mmol), triethylamine (29.01 g, 286.68 mmol) were added ) and stirred at 25°C for 0.5 hours. After the reaction was completed, water (300 mL) was added to the reaction solution, extracted with dichloromethane (200 mL×2), the organic phase was washed with saturated brine (150 mL×2), dried over anhydrous sodium sulfate, filtered, concentrated, and the residues The compound was purified by silica gel column chromatography (petroleum ether:ethyl acetate=15:1 to 5:1) to obtain compound B-4. MS-ESI calculated [M+H] ⁺ 340.8, found 340.8.

the fourth step

Compound B-4 (500 mg, 1.38 mmol) was dissolved in methanol (10 mL), and then sodium methoxide (744.99 mg, 13.79 mmol) was added to the reaction solution, followed by stirring at 70°C for 10 hours. After the reaction was completed, the reaction solution was poured into saturated aqueous ammonium chloride (10 mL), extracted with 30 mL of ethyl acetate (10 mL×3), and the combined organic phases were washed with saturated brine 30 mL (10 mL×3) After washing, it was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound B-5. MS-ESI calculated [M+H] ⁺ 308, found 308.

the fifth step

Compound B-5 (400 mg, 1.30 mmol) was dissolved in N,N-dimethylformamide (5 mL), followed by adding iodoethane (1.01 g, 6.49 mmol) and potassium carbonate ( 359.59 mg, 2.59 mmol), stirred at 60 °C for 10 hours. After the completion of the reaction, add water (10 mL) to dilute and extract with ethyl acetate 40 mL (20 mL×2). The combined organic phases were washed with saturated brine 40 mL (20 mL×2), dried over anhydrous sodium sulfate, and filtered. ,concentrate. The crude product was purified by silica gel column chromatography (petroleum ether _: ethyl acetate=10:1 to 3:1) to obtain compound B-6. MS-ESI calculated [M+H] ⁺ 337, found 337. ¹ H NMR (400 MHz, CDCl ₃ ) δ=7.53 (s, 1H), 4.80 (s, 2H), 4.51 (q, 2H), 3.97 (s, 3H), 3.68 (t, 2H), 2.79 (br t , 2H), 1.67 (br s, 9H), 1.43-1.48 (m, 3H).

Step 6

Compound B-6 (300 mg, 891.84 μmol) was dissolved in ethyl acetate (3 mL), then a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1.11 mL) was added dropwise, and the mixture was stirred at 20° C. for 0.5 hour. . After the reaction was completed, it was concentrated under reduced pressure to obtain the hydrochloride salt of compound B-7.

MS-ESI calculated [M+H] ⁺ 236, found 236.

Step 7

Compound B-7 (250 mg, 916.67 μmol, hydrochloride) and Compound B-8 (319.5 mg, 1.83 mmol) were dissolved in dichloromethane (3 mL), followed by the addition of N,N-diisopropyl Ethylamine (118.47 mg, 916.67 μmol) and sodium triethoxyborohydride (582.84 mg, 2.75 mmol) were reacted at 20° C. for 2 hours. After the reaction was completed, the reaction solution was filtered and concentrated under reduced pressure, and the residue was separated by silica gel thin-layer chromatography (petroleum ether:ethyl acetate=3:1) to obtain Intermediate B. MS-ESI calculated [M+H] ⁺ 395, found 395.

Intermediate D

first step

Intermediate A (600 mg, 2.37 mmol) and compound D-1 (511.84 mg, 1.89 mmol) were dissolved in dioxane (10 mL), followed by the addition of 1,1-bis(diphenylphosphonium)bis Ferocene palladium chloride (173.16 mg, 236.66 μmol) and potassium carbonate (981.22 mg, 7.10 mmol), the air in the solution was replaced with nitrogen three times, and the reaction solution was stirred at 80° C. for 12 hours under nitrogen protection. After the completion of the reaction, it was diluted with 20 mL of water and extracted with 60 mL of ethyl acetate (20 mL×3). The organic phase was washed with 40 mL of saturated brine (20 mL×2), dried over anhydrous sodium sulfate, filtered under reduced pressure concentrate. The residue was separated by silica gel thin layer chromatography (petroleum ether _: ethyl acetate=5:1) to obtain compound D-2. MS-ESI calculated [M+H]+317, found 317.

second step

Compound D-2 (8 g, 25.24 mmol) and compound A-2 (7.05 g, 27.76 mmol) were dissolved in dioxane (100 mL), followed by the addition of 1,1-bis(diphenylphosphonium) ) ferrocene palladium chloride (1.85 g, 2.52 mmol) and potassium acetate (7.43 g, 75.72 mmol), the air in the solution was replaced with nitrogen three times, and the reaction solution was stirred at 90° C. for 4 hours under nitrogen protection. After completion of the reaction, the reaction solution was filtered and concentrated under reduced pressure, and the obtained residue was separated by silica gel chromatography (petroleum ether:ethyl acetate=30:1 to 10:1) to obtain Intermediate D. MS-ESI calculated [M+H]+364, found 364. ¹ H NMR (400 MHz, CDCl ₃ ) δ=7.68 (dd, J=4.3, 4.9 Hz, 1H), 7.33-7.28 (m, 2H), 7.11 (t, J=7.8 Hz, 1H), 6.83 (dd, J=1.5, 8.0 Hz, 1H), 6.64 (dd, J=1.4, 7.5 Hz, 1H), 1.39 (s, 12H).

Intermediate C

first step

Compound C-1 (10 g, 49.01 mmol) was added to acetonitrile (100 mL) and water (150 mL), aqueous hydroiodic acid (100 mL, 55% by mass) was added, and the mixture was stirred at 0° C. for 0.5 hour. Sodium nitrite (50.73 g, 735.21 mmol) was dissolved in water (100 mL) and added dropwise to the above reaction solution at 0°C. After the dropwise addition, the mixture was stirred at 20° C. for 1 hour, and then continued to be stirred at 50° C. for 16 hours. After the reaction was completed, the mixture was cooled to 20°C, and added dropwise to an aqueous sodium hydroxide solution (600 mL, 20% by mass) while maintaining the temperature at 0°C. The mixture was extracted with ethyl acetate (500 mL×2), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel column chromatography (petroleum ether:ethyl acetate=100:1) to obtain compound C-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ = 8.08 (s, 1H), 4.05 (s, 3H).

second step

Compound C-2 (9.2 g, 29.22 mmol) was dissolved in tetrahydrofuran (100 mL), added dropwise to isopropylmagnesium chloride lithium chloride complex (1.3 mol/L, 40.45 mL) at minus 60°C and stirred for 30 minutes , and then N,N-dimethylformamide (6.41 g, 87.65 mmol) was added at minus 40°C and stirred for 1 hour. After the completion of the reaction, a saturated aqueous ammonium chloride solution (100 mL) was carefully added dropwise to the reaction, followed by extraction with ethyl acetate (50 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. Compound C-3 is obtained. MS-ESI calculated [M+H] ⁺ 217 and 219, found 217 and 219.

third step

Compound C-3 (1.5 g, 6.91 mmol) was dissolved in methanol (15 mL), then sodium borohydride (784.47 mg, 20.74 mmol) was added at 0°C, and the mixture was stirred at 25°C for 2 hours. After the reaction was completed, the reaction was quenched with 20 mL of saturated aqueous ammonium chloride solution, most of methanol was concentrated under reduced pressure, diluted with water (20 mL), and then extracted with ethyl acetate (90 mL), and the organic phase was washed with saturated brine (60 mL). mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (petroleum ether _: ethyl acetate=30 _: 1 to 10:1) to obtain compound C-4. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ=8.34 (s, 1H), 5.24 (t, J=6.0 Hz, 1H), 4.52 (d, J=6.0 Hz, 2H), 3.94 (s, 3H)

the fourth step

Compound C-4 (1.4 g, 6.39 mmol) and intermediate D (2.33 g, 6.39 mmol) were dissolved in dioxane (15 mL) and water (2 mL), followed by the addition of [1,1-bis (diphenylphosphine)ferrocene]palladium dichloride (467.68 mg, 639.17 mmol) and potassium carbonate (2.65 g, 19.17 mmol), the air in the solution was replaced with nitrogen three times, and the reaction solution was under nitrogen protection Stir at 65°C for 4 hours. After the reaction was completed, it was diluted with water (40 mL) and extracted with ethyl acetate (150 mL). The organic phase was washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was separated by silica gel column chromatography (petroleum ether _: ethyl acetate=30 _: 1 to 3 _: 1) to obtain compound C-5.

the fifth step

Compound C-5 (171.63 mg, 456.18 μmol) and Intermediate B (200 mg, 506.87 μmol) were dissolved in tetrahydrofuran (4 mL), and a solution of lithium bistrimethylsilylamide in tetrahydrofuran (1 mol) was added under zero nitrogen protection. /L, 1.77 mL), the reaction solution was reacted at zero for 1 hour. After the reaction, the reaction solution was quenched with saturated ammonium chloride solution (20 mL), and extracted with ethyl acetate (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified and isolated by silica gel thin layer chromatography (petroleum ether _: ethyl acetate=1 _: 2) to obtain compound C-6. MS-ESI calculated [M+H] ⁺ 724, found 724.

Step 6

Compound C-6 (130 mg, 186.27 mmol) was dissolved in 1,1-dichloroethane (2 mL), manganese dioxide (161.94 mg, 1.86 mmol) was added, and the reaction solution was heated to The reaction was carried out at 90°C for 2 hours. After the reaction was completed, it was filtered and concentrated. Intermediate C is obtained. MS-ESI calculated [M+H] ⁺ 722, found 722.

Intermediate L

first step

Compound K-1 (5.30 g, 48.38 mmol) was replaced with compound L-1 (6 g, 27.65 mmol) in the same manner as intermediate K. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain compound L-2. MS-ESI calculated [M+H] ⁺ 288 and 290, found 288 and 290.

second step

Compound K-2 (9 g, 32.83 mmol) was replaced with compound L-2 (5 g, 17.35 mmol) in the same manner as intermediate K. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain intermediate L. MS-ESI calculated [M+H] ⁺ 402 and 404, found 402 and 404.

Intermediate E

first step

Intermediate L (100 mg, 248.51 μmol) and Intermediate D (99.52 mg, 273.36 μmol) were dissolved in dioxane (5 mL) and water (0.5 mL), followed by the addition of [1,1-bis( diphenylphosphine)ferrocene]palladium dichloride (18.18 mg, 24.85 μmol) and potassium carbonate (103.04 mg, 745.53 μmol), the air in the solution was replaced with nitrogen three times, and the reaction solution was placed under nitrogen protection for 65 Stir at °C for 4 hours. After the completion of the reaction, it was diluted with 20 mL of water and extracted with 120 mL of ethyl acetate (40 mL×3). The organic phase was washed with 60 mL of saturated brine (30 mL×2), dried over anhydrous sodium sulfate, filtered under reduced pressure concentrate. The residue was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate=3:1) to obtain Intermediate E. MS-ESI calculated [M+H] ⁺ 559, found 559. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.31 (s, 1H), 7.53 (dd, J=1.7, 7.7 Hz, 1H), 7.40 (t, J=7.6 Hz, 1H), 7.26 (dd, J =1.7, 7.5Hz, 1H), 7.03 (t, J=7.8Hz, 1H), 6.81 (dd, J=1.5, 8.2Hz, 1H), 6.52 (dd, J=1.5, 7.5Hz, 1H), 4.49 (s, 2H), 3.97(s, 3H), 3.83(s, 2H), 3.48-3.43(m, 2H), 1.47(s, 3H), 1.11(s, 1H), 0.80(s, 9H), 0.00 (s, 6H).

Intermediate F

first step

Compound B-2 (1 g, 4.23 mmol) was dissolved in tetrahydrofuran (10 mL) and water (2 mL), then sodium hydroxide (845.05 mg, 21.13 mmol) was added to the reaction solution, followed by stirring at 70°C 10 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain the crude product of compound F-1. MS-ESI calculated [M+H] ⁺ 191, found 191.

second step

Compound F-1 (0.9 g, 4.73 mmol) was dissolved in ethanol (10 mL), and concentrated sulfuric acid (2.32 g, 23.66 mmol) was added to the reaction solution, followed by stirring at 70°C for 2 hours. After the reaction was completed, the filtrate was filtered and the residue concentrated under reduced pressure was purified by silica gel column chromatography (petroleum ether _: ethyl acetate=20:1 to 1:1) to obtain compound F-2. MS-ESI calculated [M+H] ⁺ 219, found 219.

third step

Compound F-2 (130 mg, 583.85 mmol) and compound F-3 (267.04 mg, 1.75 mmol), cesium carbonate (570.68 mg, 1.75 mmol) were dissolved in N,N dimethylformamide (2 mL ) and stirred at 80°C for 0.5 hours. After completion of the reaction, the residue concentrated under reduced pressure was filtered and purified by silica gel thin layer chromatography (petroleum ether:ethyl acetate=2:1) to obtain compound F-4. MS-ESI calculated [M+H] ⁺ 268, found 268.

the fourth step

Compound F-4 (90 mg, 325.48 μmol) was dissolved in glacial acetic acid (2 mL), then sodium cyanoborohydride (40.91 mg, 650.97 μmol) was added to the reaction solution, and the mixture was stirred at 20° C. for 0.5 hour. The reaction solution was concentrated under reduced pressure to obtain a crude product of compound F-5. MS-ESI calculated [M+H] ⁺ 272, found 272.

the fifth step

Compound F-5 (80 mg, 293.85 μmol) was dissolved in dichloromethane (2 mL), followed by the addition of Compound B-8 (102.44 mg, 587.70 μmol) and sodium triethoxyborohydride (186.84 mg, 881.55 μmol), the reaction solution was reacted at 20° C. for 1 hour. After the reaction was completed, the reaction solution was filtered and concentrated under reduced pressure. The residue was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate=3:1) to obtain compound F-6. MS-ESI calculated [M+H] ⁺ 431, found 431.

Step 6

Compound F-6 (1.2 g, 2.79 mmol) and compound C-5 (838.88 mg, 2.23 mmol) were dissolved in tetrahydrofuran (15 mL), and then bis(trimethyl) was added to the reaction solution at 0°C Si)lithium amide solution in tetrahydrofuran (1 mol/L, 9.75 mL) was stirred at 25°C for 1 hour. After the completion of the reaction, the reaction solution was poured into saturated aqueous ammonium chloride solution (50 mL) at 0°C, then diluted with water (30 mL), extracted with ethyl acetate (50 mL×3), and the combined organic phases were saturated After washing with brine (20 ml×2), drying with anhydrous sodium sulfate, filtration and concentration under reduced pressure, the residue was separated by silica gel column (petroleum ether:ethyl acetate=20:1 to 5:1) to obtain Intermediate F . MS-ESI calculated [M+H] ⁺ 760, found 760.

Intermediate G

first step

Compound B-4 (1 g, 2.76 mmol) was dissolved in N,N-dimethylacetamide (10 mL), followed by adding zinc cyanide (649.76 mg, 5.52 mmol) and methanesulfonic acid to the reaction solution (2-Dicyclohexylphosphine)-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl)(2-amino-1,1-biphenyl-2- base) palladium(II) (500.06 mg, 551.64 μmol), stirred at 120° C. for 10 hours under nitrogen protection. After the reaction was completed, diluted with water (10 mL), extracted with ethyl acetate 30 mL (10 mL×3), the combined organic phases were washed with saturated brine 30 mL (10 mL×3), dried over anhydrous sodium sulfate, Filter and concentrate. The obtained residue was subjected to high performance liquid chromatography (chromatographic column: Phenomenex luna C18 250*50 mm*10 microns; mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume; mobile phase B: acetonitrile; B%: 25%-55% , 30 minutes) to obtain compound G-1. MS-ESI calculated [M+H] ⁺ 332, found 332. ¹ H NMR (400 MHz, DMSO-d ₆ ) δppm 8.23 (s, 1H), 4.64 (br s, 2H), 3.66-3.72 (m, 2H), 3.02 (br s, 2H), 1.44 (s, 9H) .

second step

Compound G-1 (150 mg, 452.67 μmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (1 mL), and stirred at 20° C. for 1 hour. After the reaction was completed, it was concentrated under reduced pressure to obtain the trifluoroacetic acid salt of compound G-2. MS-ESI calculated [M+H] ⁺ 232, found 232.

third step

Compound G-2 (100 mg, 432.43 μmol, trifluoroacetate) and Compound B-8 (150.76 mg, 864.86 μmol) were dissolved in methanol (5 mL), followed by the addition of N,N-diisopropyl Ethylamine (55.89 mg, 432.43 μmol) and sodium cyanoborohydride (67.94 mg, 1.08 mmol), the reaction solution was reacted at 20° C. for 1 hour. After the reaction was completed, the reaction solution was poured into saturated aqueous ammonium chloride (30 mL), extracted with 60 mL of ethyl acetate (30 mL×2), and the combined organic phases were washed with saturated brine 60 mL (30 mL×2) After washing, it was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain intermediate G. MS-ESI calculated [M+H] ⁺ 389, found 389.

Intermediate K

first step

Compound C-3 (7 g, 32.26 mmol) and compound K-1 (5.30 g, 48.38 mmol) were dissolved in methanol (100 mL), and N,N-diisopropylethylamine (8.34 g, 64.51 mmol) was added. mmol) and stirred at 25°C for 0.5 h, then sodium cyanoborohydride (2.43 g, 38.71 mmol) was added and stirred at 25°C for 0.5 h. After the completion of the reaction, water (200 mL) was carefully added to the reaction, followed by extraction with ethyl acetate (100 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. Compound K-2 is obtained. MS-ESI calculated [M+H] ⁺ 274 and 276, found 274 and 276.

second step

Compound K-2 (9 g, 32.83 mmol) was dissolved in dichloromethane (100 mL), tert-butyldimethylsilyl chloride (12.37 g, 82.08 mmol) and imidazole (7.82 g, 114.92 mmol) were added , the mixture was stirred at 25 °C for 16 h. After completion of the reaction, water (100 mL) and dichloromethane (100 mL) were added to the reactant and stirred for 15 minutes, the organic phase was separated and dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel column chromatography (petroleum ether:ethyl acetate=10:1 to 3:1) to obtain Intermediate K. MS-ESI calculated [M+H] ⁺ 388 and 390, found 388 and 390.

Intermediate I

Intermediate K (7.4 g, 19.05 mmol) and Intermediate D (6.94 g, 19.05 mmol) were dissolved in dioxane (150 mL) and water (15 mL), followed by the addition of [1,1-bis( Diphenylphosphine)ferrocene]palladium dichloride (1.39 g, 1.91 mmol) and potassium carbonate (7.90 g, 57.16 mmol) were replaced with nitrogen three times, and the reaction solution was stirred under nitrogen protection at 65°C for 4 Hour. After the reaction was completed, it was diluted with 100 mL of water and extracted with ethyl acetate (100 mL×2). The organic phase was washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was separated by silica gel chromatography column (petroleum ether:ethyl acetate=50:1 to 5:1) to obtain Intermediate I. MS-ESI calculated [M+H] ⁺ 545, found 545. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.43 (s, 1H), 7.56 (dd, J=1.8, 7.7 Hz, 1H), 7.38 (t, J=7.6 Hz, 1H), 7.28 (dd, J= 1.8, 7.5Hz, 1H), 7.15-7.04 (m, 1H), 6.79 (dd, J=1.5, 8.1Hz, 1H), 6.66 (dd, J=1.5, 7.5Hz, 1H), 4.47 (t, J =6.2Hz, 1H), 4.15(br s, 2H), 3.99(s, 3H), 3.81-3.76(m, 2H), 3.07-2.95(m, 2H), 0.90-0.81(m, 10H), 0.01 (br s, 6H).

Intermediate J

first step

Compound B-4 (1 g, 2.93 mmol) was dissolved in dioxane (20 mL), compound A-2 (968.66 mg, 3.81 mmol), potassium acetate (719.94 mg, 7.34 mmol) and 1 were added , 1-bis(diphenylphosphoryl)ferrocene palladium(II) dichloromethane complex (239.62 mg, 293.43 μmol), the reaction solution was heated to 90° C. for 14 hours under nitrogen protection. After completion of the reaction, it was filtered, the filtrate was diluted with ethyl acetate (60 mL), and washed with saturated brine (60 mL). The organic phase was dried over sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=15:1) to obtain compound J-1. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.07 (s, 1H), 4.61 (s, 2H), 4.37-4.33 (m, 2H), 3.65-3.55 (m, 2H), 3.10-3.06 (m, 2H) ), 1.44(s, 9H), 1.35-1.31(m, 3H), 1.17(s, 12H).

second step

Compound J-1 (300 mg, 693.94 mmol) was dissolved in tetrahydrofuran (3 mL) and water (1.5 mL), sodium perborate tetrahydrate (427.08 mg, 2.78 mmol) was added, and the reaction solution was reacted at room temperature for 1 Hour. After completion of the reaction, the reaction solution was adjusted to pH 6 with 1 mol/L dilute hydrochloric acid, extracted with ethyl acetate (20 mL), the organic phase was dried over sodium sulfate, filtered and concentrated. The crude product was separated by silica gel thin layer chromatography (ethyl acetate) to give Intermediate J. MS-ESI calculated [M+H] ⁺ 323, found 323.

Example 1

first step

Compound D-1 (2 g, 7.40 mmol) was dissolved in tetrahydrofuran (20 mL), and a solution of n-butyllithium in n-hexane (3.26 mL, 2.5 mol per liter) was added at minus 78 degrees Celsius under nitrogen protection, and the reaction was half. After 1 hour, trimethyl borate (1.54 g, 14.80 mmol) was added, and the reaction solution was reacted at room temperature for 1 hour. After the completion of the reaction, the reaction solution was quenched with dilute hydrochloric acid (1 mol/L, 40 mL), and extracted with ethyl acetate (40 mL). The organic phase was dried over sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether _: ethyl acetate=10:1) to obtain compound 1-1. ¹ H NMR (400 MHz, DMSO-D ₆ ) δ = 8.42-8.46 (m, 2H), 7.70-7.73 (m, 1H), 7.36-7.39 (m, 1H), 7.22-7.24 (m, 1H).

second step

Compound 1-1 (800 mg, 3.40 mmol) was dissolved in dioxane (8 mL) and water (0.8 mL), and compound 1-2 (324.13 mg, 1.89 mmol) was added under nitrogen protection, potassium carbonate (522.16 mg, 3.78 mmol) and tetrakis(triphenylphosphonium)palladium (152.81 mg, 132.24 μmol), the reaction solution was heated to 95° C. for 3 hours under nitrogen protection. After completion of the reaction, it was filtered, the filtrate was diluted with ethyl acetate (50 mL), and washed with saturated brine (50 mL). The organic phase was dried over sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate=30:1) to obtain compound 1-3. MS-ESI calculated [M+H] ⁺ 326, found 326.

third step

Compound 1-4 (337.58 mg, 2.24 mmol) was dissolved in dichloromethane (6 mL) and ethanol (6 mL), N,N-diisopropylethylamine (482.81 mg, 3.74 mmol) and compound were added 1-3 (610 mg, 1.87 mmol) was stirred for half an hour, sodium cyanoborohydride (1.19 g, 5.60 mmol) was added, and the reaction solution was reacted at room temperature for 16 hours. After completion of the reaction, the reaction solution was diluted with dichloromethane (50 mL) and washed with saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give compound 1-5. MS-ESI calculated [M+H] ⁺ 424, found 424.

the fourth step

Compound 1-5 (550 mg, 1.29 mmol) was dissolved in dichloromethane (11 mL), di-tert-butyl dicarbonate (423.94 mg, 1.94 mmol) and triethylamine (327.59 mg, 3.24 mmol) were added , the reaction solution was reacted at room temperature for 0.5 hours. After completion of the reaction, the reaction solution was diluted with water (80 mL), extracted with dichloromethane (80 mL×2), and the organic phase was washed with saturated brine (50 mL). Dry over anhydrous sodium sulfate, filter and concentrate to obtain compound 1-6. MS-ESI calculated [M+H] ⁺ 526, found 526. ¹ H NMR (400 MHz, CD ₃ OD) δ = 7.60 (dd, J = 7.88, 1.50 Hz, 1H) 7.39 (br d, J = 1.63 Hz, 2H) 7.23-7.31 (m, 1H) 7.07-7.18 (m , 2H)4.34-4.48(m,2H)3.90-3.94(m,3H)3.83(br s,1H)3.24-3.31(m,2H)2.04-2.35(m,4H)1.52(s,9H).

the fifth step

Compound 1-6 (413 mg, 786.92 μmol) was dissolved in dioxane (8 mL) and water (0.8 mL), Intermediate A (299.26 mg, 1.18 mmol) and potassium carbonate (326.27 mg) were added , 2.36 mmol) and 1,1-bis(diphenylphosphonium)ferrocene palladium chloride (57.58 mg, 78.69 μmol). After 3 times of nitrogen replacement, the reaction solution was reacted at 85°C for 4 hours. After completion of the reaction, the reaction solution was diluted with water (40 mL), extracted with ethyl acetate (40 mL×2), and the organic phase was washed with saturated brine (10 mL). Dry over anhydrous sodium sulfate, filter and concentrate to obtain the crude product by high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150 × 40 mm × 15 μm, mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: Acetonitrile; B%: 50%-80%, 10 min) was isolated to give compound 1-7. MS-ESI calculated [M+H] ⁺ 571, found 571.

Step 6

Compound 1-7 (80 mg, 139.98 μmol) and Intermediate B (60.76 mg, 153.98 μmol) were dissolved in tetrahydrofuran (4 mL), and a solution of lithium bistrimethylsilylamide in tetrahydrofuran (1 mol) was added under zero nitrogen protection. per liter, 419.95 microliters), the reaction solution was reacted at zero for 1 hour. After the reaction was completed, the reaction solution was quenched with saturated ammonium chloride solution (30 mL), and extracted with ethyl acetate (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified and isolated by silica gel thin layer chromatography (dichloromethane:methanol=10:1) to obtain compound 1-8. MS-ESI calculated [M+H] ⁺ 919, found 919.

Step 7

Compound 1-8 (101 mg, 109.78 μmol) was dissolved in dichloromethane (2 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 2 mL) was added, and the mixture was stirred at room temperature for 1 hour. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Unisil 3-100 C18 Ultra 150*50 mm*3 microns; mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume; mobile phase B: acetonitrile; B%: 15%- 35%, 10 min) to isolate compound 1 as the formate salt. MS-ESI calculated [M+H] ⁺ 705, found 705. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.61 (d, J=8.4, 1H), 8.32-8.45 (m, 2H), 7.83-7.88 (m, 1H), 7.77 (s, 1H), 7.65- 7.70(m, 1H), 7.46-7.57(m, 2H), 7.38-7.43(m, 1H), 7.31-7.36(m, 1H), 7.13-7.21(m, 1H), 4.14-4.21(m, 2H) ), 4.04-4.12(m, 8H), 3.97-4.02(m, 1H), 3.84-3.90(m, 2H), 3.14-3.22(m, 2H), 2.94-3.09(m, 6H), 2.33-2.46 (m, 3H), 1.83-1.97 (m, 1H).

Example 2

first step

Compound 2-1 (17.90 mg, 205.47 μmol) was dissolved in dichloromethane (3 mL), N,N-diisopropylethylamine (35.41 mg, 273.96 μmol) and Intermediate C (99 mg, 136.98 μmol), after stirring for half an hour, sodium cyanoborohydride (87.10 mg, 410.94 μmol) was added, and the reaction solution was reacted at room temperature for 12 hours. After the reaction was completed, it was diluted with water (20 mL) and extracted with dichloromethane (25 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel thin layer chromatography (dichloromethane _: methanol=12:1) to obtain compound 2-2. MS-ESI calculated [M+H] ⁺ 793, found 793.

second step

Compound 2-2 (62 mg, 78.10 μmol) was dissolved in dichloromethane (3 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1.5 mL) was added, and the mixture was stirred at 25° C. for 0.5 hour. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 10%-40 %, 8 minutes) to isolate compound 2. MS-ESI calculated [M+H] ⁺ 679, found 679. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.62 (dd, J=8.25, 1.50 Hz, 1H), 8.50 (s, 1H), 7.71-7.76 (m, 2H), 7.58 (t, J=7.63 Hz) , 1H), 7.44-7.52(m, 2H), 7.18(dd, J=7.63, 1.50Hz, 1H), 4.47-4.54(m, 1H), 4.31(s, 2H), 4.11(s, 3H), 4.04(s, 3H), 3.79-3.88(m, 4H), 3.37(br s, 1H), 3.30(br d, J=5.38Hz, 1H), 3.14-3.25(m, 1H), 3.10(br d , J=11.01Hz, 1H), 2.85-2.95(m, 4H), 2.76-2.83(m, 2H), 2.20-2.31(m, 1H), 1.89-2.00(m, 1H).

Example 3

first step

Compound 3-1 (24.61 mg, 207.55 μmol) was dissolved in dichloromethane (3 mL), N,N-diisopropylethylamine (35.76 mg, 276.73 μmol) and Intermediate C (100 mg, 138.36 μmol), after stirring for half an hour, sodium cyanoborohydride (87.97 mg, 415.09 μmol) was added, and the reaction solution was reacted at room temperature for 1 hour. After the reaction was completed, it was diluted with water (20 mL) and extracted with dichloromethane (25 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel thin layer chromatography (dichloromethane _: methanol=13:1) to obtain compound 3-2. MS-ESI calculated [M+H] ⁺ 788, found 788.

second step

Compound 3-2 (83 mg, 105.22 μmol) was dissolved in dichloromethane (3 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1.5 mL) was added, and the mixture was stirred at 25° C. for 0.5 hour. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 20%-30 %, 7 minutes) to isolate compound 3. MS-ESI calculated [M+H] ⁺ 692, found 692. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.62 (dd, J=8.32, 1.19 Hz, 1H), 8.43-8.47 (m, 1H), 7.70-7.76 (m, 2H), 7.57 (t, J= 7.63Hz, 1H), 7.43-7.52 (m, 2H), 7.17 (dd, J=7.63, 1.25Hz, 1H), 4.25-4.31 (m, 2H), 4.02-4.14 (m, 8H), 3.88-3.97 (m, 4H), 3.83 (t, J=5.82Hz, 2H), 3.50-3.60 (m, 1H), 2.94-3.00 (m, 2H), 2.81-2.92 (m, 4H).

Example 4

first step

Intermediate C (80 mg, 110.69 μmol) and compound 4-1 (28.16 mg, 221.38 μmol) were dissolved in dichloromethane (2 mL), then sodium triethoxyborohydride (58.65 mg, 276.73 μmol) was added. mol) and reacted at 20°C for 2 hours. After completion of the reaction, the reaction solution was filtered and concentrated, and the crude product was purified by silica gel thin layer chromatography (dichloromethane:methanol=10:1) to obtain compound 4-2. MS-ESI calculated [M+H] ⁺ 833, found 833.

second step

Compound 4-2 (30 mg, 35.97 μmol) was dissolved in ethyl acetate (1 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 44.97 μL) was added, and the mixture was stirred at 25° C. for 0.5 hour. After the completion of the reaction, the reaction solution was filtered and concentrated, and the crude product was subjected to high performance liquid chromatography (chromatographic column: Shim-pack C18 150*25 mm*10 microns; mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume; mobile phase B: acetonitrile ; B%: 11%-41%, 10 minutes) to isolate compound 4 as the formate salt. MS-ESI calculated [M+H] ⁺ 719, found 719. ¹ H NMR (400 MHz, CD ₃ OD) δppm 8.61 (dd, 1H), 8.57 (s, 1H), 8.38-8.48 (m, 2H), 7.70-7.77 (m, 2H), 7.59 (t, 1H), 7.44-7.52(m, 2H), 7.17(dd, 1H), 4.48(s, 2H), 4.09-4.19(m, 6H), 4.04(s, 3H), 3.94(s, 2H), 3.83(t, 2H), 2.99-3.08(m, 2H), 2.89(q, 4H), 2.64(br d, 2H), 2.45(br d, 2H), 2.28-2.40(m, 2H), 2.09(br d, 2H) ).

Example 5

first step

Compound 5-1 (25.65 mg, 207.55 μmol) was dissolved in dichloromethane (3 mL), N,N-diisopropylethylamine (35.76 mg, 276.73 μmol) and Intermediate C (100 mg, 138.36 μmol), after stirring for half an hour, sodium cyanoborohydride (87.97 mg, 415.09 μmol) was added, and the reaction solution was reacted at room temperature for 1 hour. After the reaction was completed, it was diluted with water (20 mL) and extracted with dichloromethane (25 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel thin layer chromatography (dichloromethane _: methanol=13:1) to obtain compound 5-2. MS-ESI calculated [M+H] ⁺ 793, found 793.

second step

Compound 5-2 (62 mg, 78.10 μmol) was dissolved in dichloromethane (3 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1.5 mL) was added, and the mixture was stirred at 25° C. for 0.5 hour. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns; mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume; mobile phase B: acetonitrile; B%: 18%-28 %, 7 minutes) to isolate compound 5. MS-ESI calculated [M+H] ⁺ 679, found 679. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.62 (dd, J=8.28, 1.25 Hz, 1H), 8.44-8.47 (m, 1H), 7.70-7.77 (m, 2H), 7.54-7.60 (m, 1H), 7.43-7.53(m, 2H), 7.18(dd, J=7.59, 1.19Hz, 1H), 4.27-4.33(m, 2H), 3.97-4.14(m, 8H), 3.85(t, J= 5.65Hz, 2H), 3.66(s, 2H), 3.04-3.11(m, 2H), 2.89-2.96(m, 4H), 0.88-0.94(m, 2H), 0.72-0.78(m, 2H).

Example 6

first step

Intermediate C (110 mg, 152.20 μmol) was dissolved in dichloromethane (3 mL), compound 6-1 (17.15 mg, 228.30 μmol), sodium triethoxyborohydride (96.77 mg, 456.60 μmol) were added ). The reaction solution was reacted at room temperature for 2 hours. After the completion of the reaction, the reaction solution was quenched with water (10 mL), diluted with dichloromethane (5 mL), extracted with dichloromethane (10 mL×3), and the organic phase was washed with saturated brine (20 mL×2), and the solution was washed with sodium sulfate. Dry, filter and concentrate. The crude product was purified by silica gel thin layer chromatography (dichloromethane:methanol=8:1) to obtain compound 6-2 (80 mg, crude product). MS-ESI calculated [M+H] ⁺ 781, found 781.

second step

Compound 6-2 (80 mg, 102.32 μmol) was dissolved in dichloromethane (2 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1 mL) was added, and the mixture was stirred at room temperature for 0.25 hr. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Xtimate C18 150*25 mm*10 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 7%-37%, 10 minutes) to isolate the formate salt of compound 6. MS-ESI calculated [M+H] ⁺ 667, found 667. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.62 (dd, J=1.5, 8.3 Hz, 1H), 8.55 (s, 1H), 8.52-8.49 (m, 1H), 7.76-7.71 (m, 2H) , 7.59(t, J=7.6Hz, 1H), 7.53-7.45(m, 2H), 7.18(dd, J=1.5, 7.6Hz, 1H), 4.34-4.25(m, 2H), 4.13(s, 3H) ), 4.05(s, 3H), 3.86-3.79(m, 4H), 3.74(t, J=5.9Hz, 2H), 3.16(t, J=7.2Hz, 2H), 2.93-2.85(m, 4H) , 2.79(t, J=5.9Hz, 2H), 1.95(quin, J=6.6Hz, 2H).

Example 7

first step

Intermediate C (100 mg, 138.36 μmol) was dissolved in dichloromethane (3 mL), compound 7-1 (25.64 mg, 345.91 μmol), sodium triethoxyborohydride (87.98 mg, 415.09 μmol) were added ). The reaction solution was reacted at room temperature for 1 hour and then intermediate 7-1 (10.26 mg, 138.36 μmol) and sodium triethoxyborohydride (29.33 mg, 138.36 μmol) were added. The reaction solution was reacted at room temperature for 1 hour. After the completion of the reaction, the reaction solution was quenched with water (10 mL), diluted with dichloromethane (5 mL), extracted with dichloromethane (10 mL×3), and the organic phase was washed with saturated brine (20 mL×2), and the solution was washed with sodium sulfate. Dry, filter and concentrate. The crude product was purified by silica gel thin layer chromatography (dichloromethane:methanol=10:1) to obtain compound 7-2. MS-ESI calculated [M+H] ⁺ 780, found 780.

second step

Compound 7-2 (35 mg, 44.82 μmol) was dissolved in dichloromethane (2 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1 mL) was added, and the mixture was stirred at room temperature for 0.25 hr. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Xtimate C18 150*25 mm*10 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 15%-35%, 10 minutes) to isolate the formate salt of compound 7. MS-ESI calculated [M+H] ⁺ 666, found 666. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.60 (brd, J=8.4 Hz, 1H), 8.55 (s, 1H), 8.46-8.40 (m, 1H), 7.75-7.67 (m, 2H), 7.56 (brt, J=7.5Hz, 1H), 7.51-7.41 (m, 2H), 7.18-7.13 (m, 1H), 4.08 (s, 3H), 4.03 (br d, J=3.8Hz, 5H), 3.80 (br d, J=7.2Hz, 4H), 3.04 (br d, J=5.5Hz, 2H), 2.97 (br d, J=5.6Hz, 2H), 2.91-2.84 (m, 4H), 2.76 (br d, J=5.6Hz, 2H) t, J=5.7Hz, 2H), 2.67(s, 3H).

Example 8

first step

Compound 8-1 (28.21 mg, 228.30 μmol) was dissolved in dichloromethane (3 mL), and N,N-diisopropylethylamine (39.34 mg, 304.40 μmol) and Intermediate C (110 mg, 304.40 μmol) were added. 152.20 μmol), after stirring for half an hour, sodium cyanoborohydride (96.77 mg, 456.60 μmol) was added, and the reaction solution was reacted at room temperature for 12 hours. After the reaction was completed, it was diluted with water (20 mL) and extracted with dichloromethane (25 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel thin layer chromatography (dichloromethane _: methanol=10:1) to obtain compound 8-2. MS-ESI calculated [M+H] ⁺ 793, found 793.

second step

Compound 8-2 (68 mg, 85.66 μmol) was dissolved in dichloromethane (3 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1.5 mL) was added, and the mixture was stirred at 25° C. for 0.5 hour. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 10%-40 %, 7 min) to isolate compound 8 as the formate salt. MS-ESI calculated [M+H] ⁺ 679, found 679. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.58-8.65 (m, 1H), 8.49 (s, 1H) 8.43 (br s, 1H), 7.70-7.78 (m, 2H), 7.59 (t, J= 7.63Hz, 1H), 7.45-7.53(m, 2H), 7.18(dd, J=7.63, 1.38Hz, 1H), 4.52(s, 2H), 4.30-4.41(m, 3H), 4.12(s, 3H) ), 4.06(s, 3H), 3.94-4.01(m, 4H), 3.81-3.87(m, 2H), 3.35-3.37(m, 3H), 3.02-3.07(m, 2H), 2.91(q, J =5.38Hz, 4H).

Example 9

first step

Compound 3-1 (25.84 mg, 217.92 μmol) was dissolved in dichloromethane (3 mL), N,N-diisopropylethylamine (37.55 mg, 290.56 μmol) and Intermediate C (105 mg, 290.56 μmol) were added. 145.28 μmol), after stirring for half an hour, sodium cyanoborohydride (92.37 mg, 435.85 μmol) was added, and the reaction solution was reacted at room temperature for 1 hour. After the reaction was completed, it was diluted with water (20 mL) and extracted with dichloromethane (25 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel thin layer chromatography (dichloromethane _: methanol=14 _: 1) to obtain compound 9-1. MS-ESI calculated [M+H] ⁺ 788, found 788.

second step

Compound 9-1 (51 mg, 64.65 μmol) was dissolved in trifluoroacetic acid (3 mL) and stirred at 25° C. for 16 hours. After the reaction solution was detected by LC-MS, it was found that there were raw materials and the reaction was not completed. The reaction solution was heated to 30° C. and stirred for 4 hours. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 10%-40 %, 7 minutes) to isolate compound 9. MS-ESI calculated [M+H] ⁺ 674, found 674. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.60-8.63 (m, 1H), 8.42 (s, 1H), 7.83 (s, 1H), 7.72 (dd, J=7.69, 1.69 Hz, 1H), 7.55 -7.60 (m, 1H), 7.51 (t, J=7.94Hz, 1H), 7.45 (dd, J=7.57, 1.69Hz, 1H), 7.19 (dd, J=7.63, 1.63Hz, 1H), 4.30- 4.39(m, 2H), 4.07-4.10(m, 6H), 3.94(br t, J=5.38Hz, 4H), 3.81-3.90(m, 2H), 3.69(t, J=7.25Hz, 2H), 3.52-3.60 (m, 1H), 3.44 (br s, 2H), 3.25 (br d, J=4.50Hz, 2H), 3.06 (brt, J=5.88Hz, 2H).

Example 10

first step

Compound L-1 (1.72 g, 13.95 mmol) was dissolved in dichloromethane (20 mL), to the reaction solution was added compound 10-1 (2 g, 9.30 mmol), N,N-diisopropylethyl acetate Amine (3.61 g, 27.90 mmol), stirred at 25 °C for 0.5 h. Sodium triacetoxyborohydride (5.91 g, 27.90 mmol) was added to the reaction solution, and the mixture was stirred at 25°C for 2 hours. After the completion of the reaction, water (100 mL) was added to the reaction solution, extracted with dichloromethane (50 mL×3), the organic phase was washed with saturated brine (50 mL×2), and the combined organic phases were dried over anhydrous sodium sulfate. , filtered and concentrated under reduced pressure to obtain crude product. The crude product was separated by silica gel column chromatography (dichloromethane:methanol=10:1) to obtain compound 10-2. MS-ESI calculated [M+H] ⁺ 288, found 288.

second step

Compound 10-2 (1.7 g, 5.94 mmol) was dissolved in dichloromethane (20 mL). To the reaction solution were added imidazole (1.21 g, 17.82 mmol), tert-butyldimethylsilyl chloride (1.79 g, 17.82 mmol). mmol) and N,N-dimethylaminopyridine (145.15 mg, 1.19 mmol), and stirred at 45°C for 10 hours under nitrogen protection. After completion of the reaction, the reaction solution was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate=3:1) to obtain compound 10-3.

MS-ESI calculated [M+H] ⁺ 402, found 402.

third step

Compound 10-3 (1.6 g, 4.00 mmol), intermediate D (1.45 g, 4.00 mmol), potassium carbonate (1.66 g, 11.99 mmol) 1,1-bis(diphenylphosphonium)ferrocene Palladium chloride (292.37 mg, 399.58 μmol) was dissolved in dioxane (20 mL) and water (2 mL) and stirred under nitrogen at 75° C. for 10 hours. After completion of the reaction, the reaction solution was filtered and concentrated under reduced pressure to obtain a crude product, which was separated by silica gel chromatography (petroleum ether:ethyl acetate=8:1 to 3:1) to obtain compound 10-4. MS-ESI calculated [M+H] ⁺ 557, found 557.

the fourth step

Compound 10-4 (200.0 mg, 358.66 μmol) was dissolved in tetrahydrofuran (3 mL), and intermediate B (183.98 mg, 466.26 μmol), lithium bis(trimethylsilyl)amide (1 mol) were added to the reaction solution. /L n-hexane solution, 1.08 mL), stirred at 0°C for 1 hour under nitrogen protection. After the reaction, saturated ammonium chloride (20 mL) was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate=1:1) to obtain compound 10-5. MS-ESI calculated [M+H] ⁺ 905, found 905.

the fifth step

Compound 10-5 (110.0 mg, 121.39 μmol) was dissolved in dichloromethane (1 mL), followed by adding hydrogen chloride in ethyl acetate (4 mol/L, 5.5 mL) to the reaction solution, and stirring at 25° C. 0.5 hours. After the reaction was completed, concentrated, and the crude product was subjected to high performance liquid chromatography (chromatographic column: Waters Xbridge C18 150*25 mm*5 microns; mobile phase: mobile phase A: aqueous solution of 10 mmol ammonium bicarbonate by volume; mobile phase B: acetonitrile ; B%: 54%-84%, 10 minutes) to isolate compound 10. MS-ESI calculated [M+H] ⁺ 677, found 677. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.58 (dd, J=1.4, 8.3 Hz, 1H), 7.70 (s, 1H), 7.49-7.41 (m, 3H), 7.33-7.27 (m, 2H) , 7.13(dd, J=1.5, 7.6Hz, 1H), 7.05-6.97(m, 2H), 4.01(s, 3H), 3.86(s, 3H), 3.82-3.73(m, 6H), 3.40(d , J=8.9Hz, 2H), 3.21(d, J=8.6Hz, 2H), 2.86(br dd, J=4.4, 10.8Hz, 4H), 2.76(t, J=5.9Hz, 2H), 1.46( s, 3H).

Example 11

first step

Compound L-1 (858.07 mg, 6.94 μmol) was dissolved in dichloromethane (15 mL), N,N-diisopropylethylamine (1.20 g, 9.26 μmol) and compound 11-1 (1 g) were added , 4.63 mmol), and after stirring for half an hour, sodium cyanoborohydride (2.94 g, 13.89 mmol) was added, and the reaction solution was reacted at 25° C. for 14 hours. After the reaction was completed, it was diluted with dichloromethane (120 mL×2) and washed with water (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel column chromatography (ethyl acetate _: methanol=30 _: 1) to obtain compound 11-2. MS-ESI calculated [M+H] ⁺ 287, found 287.

second step

Compound 11-2 (1.1 g, 3.83 mmol) was dissolved in dichloromethane (20 mL), N,N-dimethylaminopyridine (93.60 mg, 766.14 mmol) and imidazole (1.04 g, 15.32 mmol) were added , tert-butyldimethylsilyl chloride (1.73 g, 11.49 μmol) was added to the reaction solution at 0° C. under nitrogen protection. The reaction solution was reacted at 45°C under nitrogen protection for 16 hours. LC-MS monitoring of the remaining reaction starting materials, product formation. Additional N,N-dimethylaminopyridine (93.60 mg, 766.14 μmol) and imidazole (1.04 g, 15.32 mmol) were added, and the reaction solution was added tert-butyldimethylsilyl chloride (1.73 g, 1.73 g, under nitrogen protection at 0°C). 11.49 micromoles). The reaction solution was reacted under nitrogen protection at 45°C for 14 hours. After the reaction was completed, it was diluted with water (100 mL) and extracted with dichloromethane (110 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Kromasil Eternity XT 250 × 80 mm × 10 microns, mobile phase: mobile phase A: 0.05% ammonia solution by volume; mobile phase B: acetonitrile; B%: 35%-95%, 20 minutes) to obtain the crude product, and then purified and isolated by silica gel thin layer chromatography (dichloromethane:methanol=12:1) to obtain compound 11-3. MS-ESI calculated [M+H] ⁺ 401, found 403.

third step

Compound 11-3 (90 mg, 224.21 μmol) was dissolved in dioxane (3 mL) and water (0.3 mL), Intermediate D (106.12 mg, 291.47 μmol) and potassium carbonate (92.96 mg) were added, 672.62 μmol) and 1,1-bis(diphenylphosphonium)ferrocene palladium chloride (24.61 mg, 33.63 μmol), the reaction solution was stirred at 65°C for 8 hours under nitrogen protection. After the completion of the reaction, the reaction solution was filtered, the filtrate was diluted with water (40 mL), and extracted with ethyl acetate (50 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. Compound 11-4 was isolated by silica gel thin layer chromatography (dichloromethane:methanol=12:1). MS-ESI calculated [M+H] ⁺ 558, found 558.

the fourth step

Compound 11-4 (70 mg, 125.31 μmol) and Intermediate B (64.28 mg, 162.90 μmol) were dissolved in tetrahydrofuran (2 mL), and a solution of lithium bistrimethylsilylamide in tetrahydrofuran (1 mol) was added under zero nitrogen protection. per liter, 375.93 microliters), the reaction solution was reacted at 25°C for 1 hour. After the reaction, the reaction solution was quenched with saturated ammonium chloride solution (20 mL), and extracted with ethyl acetate (25 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified and isolated by silica gel thin layer chromatography (dichloromethane _: methanol=12 _: 1) to obtain compound 11-5. MS-ESI calculated [M+H] ⁺ 907, found 906.

the fifth step

Compound 11-5 (50 mg, 55.12 μmol) was dissolved in dichloromethane (5 mL), a hydrogen chloride solution in ethyl acetate (4 mol/L, 1 mL) was added, and the reaction solution was reacted at 25° C. for 0.5 hour. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns, mobile phase: mobile phase A: 0.225% formic acid aqueous solution by volume; mobile phase B: acetonitrile; B%: 10%-40 %, 7 min) to isolate compound 11. MS-ESI calculated [M+H] ⁺ 678, found 678. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.61 (dd, J=8.31, 1.47 Hz, 1H), 8.18-8.24 (m, 1H), 7.72 (s, 1H), 7.40-7.56 (m, 5H) , 7.16(dd, J=7.58, 1.47Hz, 1H), 4.12(s, 2H), 4.04(s, 3H), 3.96(s, 3H), 3.78-3.84(m, 4H), 3.70(br d, J=9.54Hz, 2H), 3.53(br d, J=9.29Hz, 2H), 2.88(br dd, J=12.53, 4.71Hz, 4H), 2.78(t, J=5.87Hz, 2H), 1.51( s, 3H).

Example 12

first step

Intermediate C (80.0 mg, 111 μmol), 12-1 (36.2 mg, 221 μmol) was dissolved in dichloromethane (2 mL), and N,N-diisopropylethylamine ( 28.6 mg, 221 μmol), stirred at 20 °C for 1 h. Sodium triacetoxyborohydride (70.4 mg, 332 μmol) was added to the reaction solution, and the mixture was stirred at 20° C. for 2 hours. After completion of the reaction, the concentrated residue was separated and purified by silica gel thin layer chromatography (dichloromethane:methanol=10:1) to obtain compound 12-2. MS-ESI calculated [M+H] ⁺ 833, found 833.

second step

Intermediate 12-2 (30.0 mg, 29.3 μmol) was dissolved in dichloromethane (2 mL), followed by adding hydrogen chloride in ethyl acetate (4 mol/L, 500 μL) to the reaction solution, at 20°C under stirring for 0.5 hours. After the reaction was completed, concentrated, and the crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75*30 mm*3 microns; mobile phase: mobile phase A: aqueous solution of 0.225% formic acid by volume; mobile phase B: acetonitrile; B%: 10%-40%, 7 minutes) isolated compound 12. MS-ESI calculated [M+H] ⁺ 719, found 719. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.63 (dd, J=1.5, 8.3 Hz, 1H), 8.47 (s, 1H), 7.79-7.66 (m, 2H), 7.58 (t, J=7.6 Hz) , 1H), 7.52-7.43(m, 2H), 7.22-7.15(m, 1H), 4.32(s, 2H), 4.10(s, 3H), 4.05(s, 3H), 3.99(br d, J= 13.6Hz, 4H), 3.88-3.76(m, 4H), 3.01-2.84(m, 4H), 2.80(t, J=5.9Hz, 2H), 2.45-2.20(m, 4H), 1.29(s, 3H) ).

Example 13

first step

Compound B-6 (300 mg, 892 μmol), Intermediate E (334 mg, 595 μmol) was dissolved in tetrahydrofuran (6 mL), followed by adding bis(trimethylsilyl)amino to the reaction solution at 0°C Lithium (1 mol/L, 1.49 mL) was stirred at 15°C for 2 hours. After the reaction, saturated aqueous ammonium chloride solution (20 mL) was added to the reaction solution, extracted with ethyl acetate (15 mL×3), the organic phase was washed with saturated brine (20 mL×3), and dried over anhydrous sodium sulfate. , filtered, and the concentrated residue was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate=2:1) to obtain compound 13-1. MS-ESI calculated [M+H] ⁺ 849, found 849.

second step

Compound 13-1 (200 mg, 235.32 μmol) was dissolved in dichloromethane (3 mL), then trifluoroacetic acid (924.00 mg, 8.10 mmol, 0.6 mL) was added, and the reaction solution was reacted at 15° C. for 0.5 h . After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain compound 13-2. MS-ESI calculated [M+H] ⁺ 749, found 749.

third step

Compound 13-2 (100 mg, 133.37 μmol) and lithium perchlorate (28.38 mg, 266.74 μmol) were dissolved in ethanol (1.5 mL), then compound 13-3 (28.85 mg, 400.11 μmol) was added, The reaction solution was placed in a sealed tube and stirred at 80°C for 6 hours. After the reaction was completed, it was concentrated under reduced pressure, and the residue was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate=1:1) to obtain compound 13-4. MS-ESI calculated [M+H] ⁺ 821, found 821.

the fourth step

Compound 13-4 (90 mg, 109.50 μmol) was dissolved in dichloromethane (1 mL), and an ethyl acetate solution of hydrogen chloride (4 mol/L, 0.2 mL) was added, and the reaction solution was reacted at 25° C. for 1 hour . After the completion of the reaction, concentrated under reduced pressure to obtain the residue by high performance liquid chromatography (chromatographic column: Phenomenex luna C18 150*25 mm*10 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B %: 7%-37%, 10 min) to isolate compound 13 as the formate salt. MS-ESI calculated [M+H] ⁺ 707, found 707. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.63 (dd, J=1.4, 8.3 Hz, 1H), 8.50 (s, 1H), 8.44 (s, 1H), 7.78-7.68 (m, 2H), 7.59 (t, J=7.7Hz, 1H), 7.53-7.41 (m, 2H), 7.17 (dd, J=1.4, 7.7Hz, 1H), 4.59 (s, 2H), 4.19 (d, J=10.9Hz, 2H), 4.13(s, 3H), 4.08-3.99(m, 5H), 3.94(s, 2H), 3.07-2.94(m, 2H), 2.91-2.80(m, 2H), 2.61(s, 2H) , 1.58(s, 3H), 1.26(s, 6H).

Example 14

first step

Compound 14-1 (43.45 mg, 345.91 μmol) was dissolved in dichloromethane (3 mL), N,N diisopropylethylamine (53.65 mg, 415.09 μmol) was added, Intermediate C (100 mg, 138.36 micromoles). The reaction solution was stirred at room temperature for 0.5 hours and then sodium triethoxyborohydride (87.98 mg, 415.09 μmol) was added. The reaction solution was reacted at room temperature for 1 hour. After the completion of the reaction, the reaction solution was quenched with water (10 mL), diluted with dichloromethane (5 mL), extracted with dichloromethane (10 mL×3), and the organic phase was washed with saturated brine (20 mL×2), and the solution was washed with sodium sulfate. Dry, filter and concentrate. The crude product was purified by silica gel thin-layer chromatography (dichloromethane _: methanol=10 _: 1) to obtain compound 14-2. MS-ESI calculated [M+H] ⁺ 795, found 795.

second step

Compound 14-2 (90 mg, 113.08 μmol) was dissolved in dichloromethane (2 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 1 mL) was added, and the mixture was stirred at room temperature for 0.25 hr. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Xtimate C18 150*25 mm*10 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 9%-39%, 10 minutes) to isolate compound 14. MS-ESI calculated [M+H] ⁺ 681, found 681. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.62 (dd, J=1.5, 8.3 Hz, 1H), 8.55 (s, 1H), 8.52-8.49 (m, 1H), 7.76-7.71 (m, 2H) , 7.59(t, J=7.6Hz, 1H), 7.53-7.45(m, 2H), 7.18(dd, J=1.5, 7.6Hz, 1H), 4.34-4.25(m, 2H), 4.13(s, 3H) ), 4.05(s, 3H), 3.86-3.79(m, 4H), 3.74(t, J=5.9Hz, 2H), 3.16(t, J=7.2Hz, 2H), 2.93-2.85(m, 4H) , 2.79(t, J=5.9Hz, 2H), 1.95(quin, J=6.6Hz, 2H).

Example 15

first step

Intermediate F (150 mg, 197.18 μmol) was dissolved in 1,2-dichloroethane (2 mL), manganese dioxide (171.42 mg, 1.97 mmol) was added, and the reaction solution was heated to 90°C under nitrogen protection , manganese dioxide (171.42 mg, 1.97 mmol) was added after the reaction for 2 hours. The reaction solution was heated to 90°C under nitrogen protection for 2 hours. After the reaction was completed, it was filtered and concentrated. Compound 15-1 was obtained. MS-ESI calculated [M+H] ⁺ 758, found 758.

second step

Compound 2-1 (34.21 mg, 276.78 μmol) was dissolved in dichloromethane (3 mL), N,N diisopropylethylamine (35.77 mg, 276.78 μmol), compound 15-1 (140 mg, 184.52 micromoles), stirred at room temperature for 0.5 hours and then added sodium triethoxyborohydride (114.37 mg, 539.61 micromoles). The reaction solution was reacted at room temperature for 1 hour. After the completion of the reaction, the reaction solution was quenched with water (10 mL), diluted with dichloromethane (5 mL), extracted with dichloromethane (10 mL×3), and the organic phase was washed with saturated brine (20 mL×2), and the solution was washed with sodium sulfate. Dry, filter and concentrate. The crude product was purified by silica gel thin layer chromatography (dichloromethane:methanol=10:1) to isolate intermediate 15-2. MS-ESI calculated [M+H] ⁺ 829, found 829.

third step

Compound 15-2 (75 mg, 90.38 μmol) in dichloromethane (2 mL) was added with an ethyl acetate solution of hydrogen chloride (4 mol/L, 1 mL), and the mixture was stirred at room temperature for 0.25 hr. After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Xtimate C18 150*25 mm*10 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 7%-37%, 10 minutes) to isolate compound 15. MS-ESI calculated [M+H] ⁺ 715, found 715. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.59 (dd, J=1.5, 8.3 Hz, 1H), 8.46 (s, 1H), 7.82 (s, 1H), 7.74-7.68 (m, 1H), 7.56 (t, J=7.6Hz, 1H), 7.51-7.42 (m, 2H), 7.29-7.22 (m, 1H), 7.20-7.14 (m, 1H), 7.34-6.95 (m, 1H), 4.49-4.36 (m, 1H), 4.11 (s, 2H), 4.08 (s, 3H), 3.91-3.85 (m, 2H), 3.83-3.76 (m, 2H), 3.20-3.07 (m, 2H), 3.01-2.84 (m, 6H), 2.81-2.74 (m, 2H), 2.28-2.12 (m, 1H), 1.89-1.78 (m, 1H).

Example 16

first step

Compound 16-1 (500 mg, 2.85 mmol) was dissolved in N,N-dimethylformamide (6 mL), potassium carbonate (472 mg, 3.42 mmol), iodoethane (578 mL) were added to the reaction solution. mg, 3.71 mmol) and stirred at 50°C for 2 hours. After the completion of the reaction, add water (50 mL) to dilute, extract with ethyl acetate (15 mL×3), wash the organic phase with aqueous sodium bicarbonate solution (20 mL×3), and then wash the organic phase with saturated brine (20 mL×3) Washed, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 16-2. MS-ESI calculated [M+H] ⁺ 204, found 204. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.51-8.35 (m, 1H), 7.09 (dd, J=5.7, 8.0 Hz, 1H), 4.47 (q, J=7.2 Hz, 2H), 1.42 (t, J=7.2Hz, 3H).

second step

Compound 16-2 (12.5 g, 61.40 mmol) was dissolved in N,N-dimethylformamide (80 mL), cesium acetate (12.96 g, 67.54 mmol) was added to the reaction solution, nitrogen was replaced three times, and then Stir at 80°C for 2 hours under nitrogen protection. The remaining raw materials were detected by TLC, and cesium acetate (7.07 g, 36.84 mmol) was added to the reaction solution, replaced with nitrogen three times, and then stirred at 80° C. for 2 hours under nitrogen protection. Water (400 mL) was added to dilute, extracted with ethyl acetate (200 mL×3), the organic phase was washed with saturated brine (200 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was filtered through a silica gel column. Compound 16-3 was obtained after purification and separation (petroleum ether:ethyl acetate=100:1 to 10:1). MS-ESI calculated [M+H] ⁺ 202, found 202.

third step

Compound 16-3 (7 g, 34.72 mmol) and compound F-3 (10.59 g, 69.44 mmol) were dissolved in N,N dimethylformamide (70 mL), followed by potassium carbonate (14.40 g, 104.16 g) mmol), the reaction solution was stirred at 80 °C for 2 hours. 90 mL of water was added to the reaction solution to dilute, and then extracted with ethyl acetate (60 mL×3). The combined organic phases were washed with saturated brine (30 mL×3) and dried over anhydrous sodium sulfate. The crude product obtained by filtration and concentration under reduced pressure was purified and isolated by silica gel column chromatography (petroleum ether:ethyl acetate=30:1 to 10:1) to obtain compound 16-4. MS-ESI calculated [M+H] ⁺ 252, found 252. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.62-8.33 (m, 1H), 7.19-7.03 (m, 1H), 6.95-6.40 (m, 1H), 4.65-4.28 (m, 2H), 1.50-1.26 (m, 3H).

the fourth step

Compound 16-4 (1.6 g, 6.36 mmol) was dissolved in N,N dimethylacetamide (16 mL), zinc cyanide (1.49 g, 12.72 mmol) was added, methanesulfonic acid (2-dicyclohexyl Phosphine)-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II) (1.15 g, 1.27 mmol). After the reaction solution was replaced with nitrogen three times, it was stirred at 105° C. for 12 hours under nitrogen protection. Filter and add water (50 mL), extract with ethyl acetate (40 mL×3), combine the organic phases, wash with saturated brine (30 mL×3), and dry over anhydrous sodium sulfate. The crude product obtained by filtration and concentration under reduced pressure was purified and isolated by silica gel column chromatography (petroleum ether:ethyl acetate=50:1 to 10:1) to obtain compound 16-5. MS-ESI calculated [M+H] ⁺ 243, found 243. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.75 (d, J=5.7 Hz, 1H), 7.42 (td, J=1.3, 5.7 Hz, 1H), 7.01-6.41 (m, 1H), 4.55 (q, J=7.2Hz, 2H), 1.48(t, J=7.2Hz, 3H).

the fifth step

Compound 16-5 (1.1 g, 4.54 mmol) was dissolved in methanol (40 mL), Raney nickel (389.14 mg, 454.21 μmol, 10% purity) was added under nitrogen protection, and the reaction solution was placed in a hydrogen atmosphere (pressure 50 psi) It was heated to 50°C for 12 hours. After the reaction was completed, it was filtered and concentrated. The crude product was slurried with petroleum ether:ethyl acetate=4:1 (25 mL), filtered, and the filter cake was dried to obtain compound 16-6. MS-ESI calculated [M+H] ⁺ 201, found 201. ¹ H NMR (400MHz, CDCl ₃ -d) δ=8.72 (d, J=5.7Hz, 1H), 7.67-7.36 (m, 1H), 7.25 (s, 1H), 7.18 (d, J=5.7Hz, 1H), 6.67(br s, 1H), 4.57(s, 2H).

Step 6

Compound 16-6 (780 mg, 3.90 mmol) was dissolved in dichloromethane (10 mL), and di-tert-butyl dicarbonate (935.62 mg, 4.29 mmol) and 4-dimethylaminopyridine (71.42 mg, 584.58 mmol) were added. micromoles). The reaction solution was reacted at 25°C for 0.5 hour. The crude product obtained by concentrating the reaction solution under reduced pressure was purified and isolated by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 5:1) to obtain compound 16-7. MS-ESI calculated [M+H] ⁺ 301, found 301.

Step 7

Compound 16-7 (900 mg, 3.00 mmol) was dissolved in tetrahydrofuran (10 mL), diisobutylaluminum hydride (1 mol/L, 8.99 mL) was added at minus 5 °C, and the reaction solution was reacted at 20 °C for 2 hours . Sodium sulfate decahydrate (20 g) was added and the reaction was quenched by stirring for 30 minutes, filtered, and the filtrate was concentrated to give compound 16-8. MS-ESI calculated [M+H] ⁺ 303, found 303.

Step 8

Compound 16-8 (800 mg, 2.65 mmol) was dissolved in acetic acid (8 mL), sodium cyanoborohydride (166.32 mg, 2.65 mmol) was added, and the reaction solution was reacted at 15° C. for 1 hour. The reaction solution was slowly added dropwise to saturated aqueous sodium bicarbonate solution (50 mL), stirred for 20 minutes, extracted with ethyl acetate (40 mL×3), and the combined organic phases were washed with saturated brine (30 mL×2) , and dried over anhydrous sodium sulfate. The crude product obtained by filtration and concentration under reduced pressure was purified and isolated by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 5:1) to obtain compound 16-9. MS-ESI calculated [M+H] ⁺ 287, found 287.

Step 9

Compound 16-9 (270 mg, 943.15 μmol) was dissolved in dichloromethane (3 mL), m-chloroperoxybenzoic acid (406.90 mg, 1.89 mmol, 80% purity) was added, and the mixture was stirred at 15° C. for 4 hours. 15 mL of saturated aqueous sodium sulfite solution was added at 0°C to quench the reaction, and the pH was adjusted to 7 with saturated aqueous sodium bicarbonate solution, extracted with dichloromethane (30 mL × 3), and the combined organic phases were washed with saturated brine (30 mL × 2 ) and dried over anhydrous sodium sulfate. The crude product obtained by filtration and concentration under reduced pressure was slurried with petroleum ether:methyl tert-butyl ether=1:1 (10 mL), filtered, and the filter cake was dried to obtain compound 16-10. MS-ESI calculated [M+H] ⁺ 303, found 303. ¹ H NMR (400 MHz, CDCl ₃ ) δ=8.14 (d, J=7.2 Hz, 1H), 7.11 (br d, J=7.2 Hz, 1H), 6.88-6.33 (m, 1H), 4.93-4.72 (m , 4H), 1.53(s, 9H).

Step 10

Compound 16-10 (75 mg, 248.12 mmol) was dissolved in N,N dimethylformamide (10 mL), oxalyl chloride (725.02 mg, 5.71 mmol) was slowly added dropwise at 0 °C, and the temperature was raised to 30 °C. Stir for 12 hours. The reaction solution was added dropwise to 50 mL of saturated aqueous sodium bicarbonate solution at 0°C to quench the reaction, extracted with ethyl acetate (30 mL×3), and the combined organic phases were washed with saturated brine (20 mL×2), and dried with anhydrous sodium sulfate. The crude product obtained by filtration and concentration under reduced pressure was purified by silica gel thin layer chromatography (petroleum ether:ethyl acetate=3:1) to obtain compound 16-11. MS-ESI calculated [M+H] ⁺ 321, found 321.

Step 11

Compound 16-11 (30 mg, 93.54 μmol) was dissolved in methanol (5 mL) and N,N dimethylformamide (1 mL), triethylamine (28.40 mg, 280.62 μmol) and 1,1 were added -Bis(diphenylphosphorus)ferrocene palladium dichloride (6.84 mg, 9.35 μmol), the reaction solution was heated to 80 degrees under carbon monoxide (pressure 50 psi) for 12 hours. The solution was concentrated by filtration, diluted with 10 mL of ethyl acetate and 20 mL of water, then extracted with ethyl acetate (30 mL×2), the organic phase was washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate, filtered and Concentrated under reduced pressure, the obtained crude product was purified by silica gel thin layer chromatography (petroleum ether:ethyl acetate=1:1) to obtain compound 16-12. MS-ESI calculated [M+H] ⁺ 345, found 345.

Step 12

Compound 16-12 (30 mg, 87.13 μmol) and intermediate E (43.88 mg, 78.42 μmol) were dissolved in tetrahydrofuran (2 mL), and a solution of lithium bistrimethylsilylamide in tetrahydrofuran (1 mol/L, 304.96 μL), the reaction solution was reacted at 15° C. for 1 hour. The reaction solution was quenched by adding saturated ammonium chloride solution (20 mL) at 0°C, and extracted with ethyl acetate (20 mL×3). The combined organic phases were washed with saturated brine (15 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained crude product was purified by silica gel thin layer chromatography (petroleum ether:ethyl acetate=1:1) Compounds 16-13 were obtained. MS-ESI calculated [M+H] ⁺ 871, found 871.

step thirteen

Compound 16-13 (32 mg, 36.7 μmol) was dissolved in dichloromethane (2 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 432.43 μL) was added, and the mixture was stirred at 15° C. for 6 hours. The reaction solution was concentrated to obtain the hydrochloride salt of compound 16-14. MS-ESI calculated [M+H] ⁺ 657, found 657.

Step 14

Compound 16-14 (25 mg, 36.03 μmol) was dissolved in methanol (2 mL), N,N-diisopropylethylamine (4.66 mg, 36.03 μmol) and compound B-8 (15.70 mg, 90.06 μg) were added μmol), stirred at 15° C. for 0.5 hours, and then added sodium cyanoborohydride (6.79 mg), and the reaction solution was reacted at 15° C. for 1.5 hours. The reaction solution was diluted by adding 10 mL of water, and extracted with dichloromethane (10 mL×3). The combined organic phases were washed with saturated brine (10 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 16-15. MS-ESI calculated [M+H] ⁺ 815, found 815.

Step 15

Compound 16-15 (29 mg, 35.55 μmol) was dissolved in ethyl acetate (1 mL), and an ethyl acetate solution of hydrogen chloride (0.5 mL, 4 mol/L) was added, and the reaction solution was reacted at 15° C. for 0.5 hour. The crude product obtained by concentrating the reaction solution under reduced pressure was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Synergi C18 150*25 mm*10 microns; mobile phase: mobile phase A: volume fraction of 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B% : 8%-38%, 10 min) to isolate compound 16. MS-ESI calculated [M+H] ⁺ 701, found 701. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.58 (dd, J=1.5, 8.3 Hz, 1H), 8.46 (s, 1H), 7.91 (s, 1H), 7.70 (dd, J=1.6, 7.7 Hz , 1H), 7.56(t, J=7.6Hz, 1H), 7.52-7.06(m, 4H), 4.45(br s, 2H), 4.16(br d, J=10.9Hz, 4H), 4.09(s, 3H), 4.06 (br d, J=8.5Hz, 2H), 3.93-3.83 (m, 2H), 3.77 (t, J=5.7Hz, 2H), 2.97 (t, J=5.7Hz, 2H), 1.54 (s, 3H).

Example 17

first step

Intermediate G (400.0 mg, 1.03 mmol) was dissolved in tetrahydrofuran (2.5 mL), compound C-5 (347.68 mg, 924.11 μmol), bis(trimethylsilyl)amino was added to the reaction solution at 0°C Lithium (1 mol/L in n-hexane, 3.08 mL) was stirred at 20°C for 1 hour. After the reaction was completed, water (50 mL) was added to the reaction solution, extracted with dichloromethane (50 mL×3), the organic phase was washed with saturated brine (30 mL×3), and the combined organic phases were dried over anhydrous sodium sulfate. , filtered and concentrated under reduced pressure to obtain crude product. The crude product was separated by silica gel column chromatography (petroleum ether:ethyl acetate=20:1 to 1:1) to obtain compound 17-1. MS-ESI calculated [M+H] ⁺ 719, found 719.

second step

Compound 17-1 (200.0 mg, 277.88 μmol) was dissolved in dichloroethane (3 mL), active manganese dioxide (241.59 mg, 2.78 μmol) was added, and the mixture was stirred at 90° C. for 2 hours. After completion of the reaction, filter and concentrate under reduced pressure to obtain compound 17-2. MS-ESI calculated [M+H] ⁺ 717, found 717.

third step

Compound 17-2 (70.0 mg, 97.53 μmol) was dissolved in dichloromethane (3 mL). To the reaction solution was added compound 12-1 (23.94 mg, 146.30 μmol), N,N-diisopropylethylamine (37.82 mg, 292.60 μmol) and stirred at 25° C. for 0.5 h. Sodium triacetoxyborohydride (62.01 mg, 292.60 μmol) was added to the reaction solution, and the mixture was stirred at 25° C. for 10 hours. After the reaction was completed, water (5.0 mL) was added to the reaction solution, extracted with dichloromethane 24 mL (8 mL×3), the organic phase was washed with saturated brine (10 mL×2), and the organic phases were combined with anhydrous sulfuric acid. Dry over sodium, filter, and concentrate under reduced pressure to give crude product. The crude product was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate:ethanol=4:3:1) to obtain compound 17-3. MS-ESI calculated [M+H] ⁺ 828, found 828.

the fourth step

Compound 17-3 (50.0 mg, 60.32 μmol) was dissolved in trifluoroacetic acid (2 mL) and stirred at 25° C. for 12 hours. After the reaction was completed, concentrated, and the crude product was subjected to high performance liquid chromatography (chromatographic column: Unisil 3-100 C18 Ultra 150*50 mm*3 microns; mobile phase: mobile phase A: aqueous solution of 0.225% formic acid by volume; mobile phase B: acetonitrile ; B%: 8%-38%, 10 minutes) to isolate compound 17. MS-ESI calculated [M+H] ⁺ 714, found 714. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.63 (dd, J=1.5, 8.3 Hz, 1H), 8.47 (s, 1H), 7.79-7.66 (m, 2H), 7.58 (t, J=7.6 Hz, 1H), 7.52-7.43(m, 2H), 7.22-7.15(m, 1H), 4.32(s, 2H), 4.10(s, 3H), 4.05(s, 3H), 3.99(br d, J=13.6 Hz, 4H), 3.88-3.76(m, 4H), 3.01-2.84(m, 4H), 2.80(t, J=5.9 Hz, 2H), 2.45-2.20(m, 4H), 1.29(s, 3H) .

Example 18

first step

Compound 17-2 (60.0 mg, 83.60 μmol) was dissolved in dichloromethane (3 mL), and compound 2-1 (15.50 mg, 125.40 μmol), N,N-diisopropylethyl acetate was added to the reaction solution. Amine (32.41 mg, 250.80 μmol), stirred at 25° C. for 0.5 h. Sodium triacetoxyborohydride (53.15 mg, 250.80 μmol) was added to the reaction solution, and the mixture was stirred at 25° C. for 2 hours. After the completion of the reaction, water (5 mL) was added to the reaction solution, extracted with dichloromethane (8 mL×3), the organic phase was washed with saturated brine (10 mL×2), and the combined organic phases were dried over anhydrous sodium sulfate , filtered and concentrated under reduced pressure to obtain crude product. The crude product was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate:ethanol=4:3:1) to obtain compound 18-1. MS-ESI calculated [M+H] ⁺ 788, found 788.

second step

Compound 18-1 (21.0 mg, 26.62 μmol) was dissolved in trifluoroacetic acid (2 mL) and stirred at 25° C. for 2 hours. After the reaction was completed, concentrated, and the crude product was subjected to high performance liquid chromatography (chromatographic column: Unisil 3-100 C18 Ultra 150*50 mm*3 microns; mobile phase: mobile phase A: aqueous solution of 0.225% formic acid by volume; mobile phase B: acetonitrile ; B%: 8%-38%, 10 minutes) to isolate compound 18. MS-ESI calculated [M+H] ⁺ 674, found 674. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.56 (dd, J=1.1, 8.3 Hz, 1H), 8.51 (s, 1H), 8.34 (s, 1H), 7.73 (dd, J=1.4, 7.6 Hz) , 1H), 7.57 (t, J=7.6Hz, 1H), 7.53-7.42 (m, 2H), 7.19 (dd, J=1.4, 7.6Hz, 1H), 4.60-4.56 (m, 3H), 4.44 ( br s, 2H), 4.10(s, 3H), 3.95(s, 2H), 3.79(t, J=5.6Hz, 2H), 3.19(br t, J=5.6Hz, 4H), 3.02-2.95(m , 2H), 2.80 (t, J=5.7Hz, 2H), 2.34-2.19 (m, 1H), 2.35-2.15 (m, 1H), 2.09-1.92 (m, 1H)

Example 19

first step

Compound 6-1 (15.70 mg, 209.00 μmol) was dissolved in dichloromethane (3 mL), compound 17-2 (100 mg, 139.33 μmol), sodium triethoxyborohydride (88.59 mg, 417.99 μmol) were added mol), the reaction solution was reacted at room temperature for 2 hours. After completion of the reaction, add water (20 mL) to dilute, and extract with dichloromethane (25 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was separated by silica gel thin layer chromatography (dichloromethane:methanol=6:1) to obtain compound 19-1. MS-ESI calculated [M+H] ⁺ 776, found 776.

second step

Compound 19-1 (70 mg, 90.11 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (6 mL) was added, and the mixture was stirred at 20° C. for 12 hours. LC-MS monitored the presence of raw materials, and the reaction solution was warmed to Stir at 30°C for 3 hours. After the reaction was completed, it was concentrated. The crude product was added to a solution of tetrahydrofuran (2 mL) and saturated aqueous sodium bicarbonate solution (2 mL), stirred at 20° C. for 3 hours, diluted with water (20 mL), and extracted with dichloromethane (20 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns; mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 10%-40 %, 7 min) to isolate compound 19. MS-ESI calculated [M+H] ⁺ 662, found 662. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.58 (d, J=8.31 Hz, 1H), 8.51 (s, 1H), 8.33-8.40 (m, 1H), 7.74 (dd, J=7.76, 0.92 Hz , 1H), 7.56-7.60(m, 1H), 7.45-7.54(m, 2H), 7.18-7.24(m, 1H), 4.33(s, 2H), 4.12-4.14(m, 3H), 3.96(s) , 2H), 3.81(t, J=5.62Hz, 2H), 3.75(t, J=5.81Hz, 2H), 3.17-3.22(m, 4H), 2.98-3.04(m, 2H), 2.82(t, J=5.62Hz, 2H), 1.96 (quin, J=6.48Hz, 2H).

Example 20

first step

Intermediate J (400 mg, 1.24 mmol) was dissolved in acetone (4 mL), 2-iodopropane (1.05 mg, 6.20 mmol), potassium carbonate (342.99 mg, 2.48 mmol) were added, and the reaction solution was heated at 80 °C The reaction was continued for 4 hours. After the reaction was completed, it was diluted with ethyl acetate (60 mL×2), and extracted with water (60 mL). The organic phase was dried over sodium sulfate, filtered and concentrated. The residue was separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate=5:1) to obtain compound 20-1. MS-ESI calculated [M+H] ⁺ 365, found 365 ¹ H NMR (400 MHz, CDCl ₃ ) δppm=7.48-7.50 (m, 1H), 4.71-4.84 (m, 3H), 4.50 (q, J =7.15Hz, 2H), 3.62-3.71(m, 2H), 2.78(brt, J=5.71Hz, 2H), 1.40-1.50(m, 18H).

second step

Compound 20-1 (535 mg, 1.47 μmol) was dissolved in dichloromethane (6 mL), a solution of hydrogen chloride in ethyl acetate (4 mol/L, 3.5 mL) was added, and the reaction solution was reacted at room temperature 25° C. for 1 hour . After the completion of the reaction, the reaction solution was concentrated to obtain the crude compound 20-2. MS-ESI calculated [M+H] ⁺ 265.2, found 265.2.

third step

Compound 20-2 (440 mg, 1.46 μmol) was dissolved in methanol (24 mL), N,N-diisopropylethylamine (189.06 mg, 1.46 μmol) and compound B-8 (637.49 mg, 3.66 mg) were added mmol), after stirring for half an hour, sodium cyanoborohydride (275.79 mg, 4.39 mmol) was added, and the reaction solution was reacted at 25° C. for 16 hours. After the reaction was completed, it was diluted with ethyl acetate (60 mL×2) and washed with water (40 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel thin layer chromatography (petroleum ether:ethyl acetate=5:1) to obtain compound 20-3. MS-ESI calculated [M+H] ⁺ 423.4, found 423.4. ¹ H NMR (400 MHz, CDCl ₃ ) δ=7.38 (s, 1H), 4.58-4.74 (m, 1H), 4.39 (q, J=7.05 Hz, 2H), 3.64-3.93 (m, 4H), 2.50- 2.94(m, 6H), 1.32-1.37(m, 4H), 1.30(d, J=6.00Hz, 6H), 0.83(s, 9H), 0.00(s, 6H).

the fourth step

Compound 20-3 (120 mg, 283.93 μmol) and intermediate 1 (123.93 mg, 227.15 μmol) were dissolved in tetrahydrofuran (2 mL), and a solution of lithium bistrimethylsilylamide in tetrahydrofuran (1 mol) was added under zero nitrogen protection. /L, 851.80 μL), the reaction solution was reacted at zero for 1 hour. After the reaction, the reaction solution was quenched with saturated ammonium chloride solution (20 mL), and extracted with ethyl acetate (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified and isolated by silica gel thin layer chromatography (dichloromethane:methanol=10:1) to obtain compound 20-4. MS-ESI calculated [M+H] ⁺ 922, found 921.

the fifth step

Compound 20-4 (100 mg, 85.85 μmol) was dissolved in dichloromethane (5 mL), and an ethyl acetate solution of hydrogen chloride (4 mol/L, 4 mL) was added, and the reaction solution was reacted at room temperature at 25° C. for 0.5 hours . After the reaction was completed, it was concentrated. The crude product was subjected to high performance liquid chromatography (chromatographic column: Phenomenex Gemini-NX C18 75 × 30 mm × 3 microns, mobile phase: mobile phase A: volume fraction 0.225% formic acid aqueous solution; mobile phase B: acetonitrile; B%: 12%-42 %, 7 min) to isolate compound 20 as the formate salt. MS-ESI calculated [M+H] ⁺ 693, found 693. ¹ H NMR (400 MHz, CD ₃ OD) δ=8.61 (dd, J=8.25, 1.41 Hz, 1H), 8.44-8.52 (m, 2H), 7.70-7.74 (m, 2H), 7.56-7.61 (m, 1H), 7.45-7.53(m, 2H), 7.17(dd, J=7.64, 1.41Hz, 1H), 4.94(br s, 1H), 4.65-4.71(m, 1H), 4.55(s, 2H), 4.38-4.45(m, 2H), 4.12(s, 3H), 3.90-3.98(m, 4H), 3.84(t, J=5.75Hz, 2H), 3.00-3.07(m, 2H), 2.85-2.94( m, 4H), 1.45 (d, J=5.99Hz, 6H).

Experimental Example 1: PD-1/PD-L1 Homogenouse Time-Resolved Fluorescence (HTRF) combined with the experimental principle:

Small molecule compounds can competitively inhibit the binding of PD-1 and PD-L1 by binding to PD-L1; when the PD-1 molecule as the donor is very close to the PD-L1 molecule as the acceptor, the donor molecule will The energy is transferred to the receptor molecule, which in turn causes the receptor molecule to emit fluorescence; by detecting the intensity of fluorescence, the ability of small molecules to prevent the binding of PD-L1 to PD-1 can be tested. A homogeneous time-resolved fluorescence (HTRF) binding assay was used to detect the ability of the compounds of the present invention to inhibit the mutual binding of PD-1/PD-L1.

Experimental Materials:

PD-1/PD-L1TR-FRET detection kit was purchased from BPS Biosciences. Nivo Multilabel Analyzer (PerkinElmer).

experimental method:

Dilute PD1-Eu, Dye-labeled acceptor, PD-L1-biotin and test compound with the buffer in the kit. The compound to be tested was diluted 5-fold to the 8th concentration with a row gun, that is, from 40 μM to 0.5 nM, and the DMSO concentration was 4%, and a double-well experiment was set up. Add 5μM inhibitor to each concentration gradient of the microplate, wherein the maximum (Max) signal well and the minimum (Min) signal well add 5μL buffer containing 4% DMSO, 5μL PD-L1-biotin (PD-L1-biotin) ) (60nM), only 5μL of buffer was added to the minimum (Min) signal well, and incubated at 25°C for 20 minutes. After the incubation, 5 μL of diluted PD1-Eu (10 nM) and 5 μL of diluted dye-labeled acceptor (Dye-labeled acceptor) were added to each well. The reaction system was placed at 25 degrees for 90 minutes. After the reaction, the TR-FRET signal was read by a multi-label analyzer.

data analysis:

Using the equation (sample-Min)/(Max-Min)×100% to convert the raw data into inhibition rate, the IC ₅₀ value can be obtained by curve fitting with four parameters (log(inhibitor) vs.response in GraphPad Prism --Variable slope mode). Table 1 provides the inhibitory activity of the compounds of the examples of the present invention on PD1/PD-L1 binding.

Table 1 Test results of IC ₅₀ values of the compounds of the examples of the present invention for binding to PD-1/PD-L1

受试化合物test compound	IC ₅₀(nM) _IC50 (nM)
化合物2Compound 2	1.491.49
化合物8的甲酸盐Formate salt of compound 8	4.614.61
化合物12Compound 12	3.133.13
化合物15Compound 15	4.204.20

Experimental conclusion: The compound of the present invention has a significant inhibitory effect on the binding of PD-1/PD-L1.

Experimental Example 2: Using MDA-MR-231 cells to detect the effect of compounds on the expression level of PD-L1

Experimental principle:

The use of a triple-negative breast cancer cell line (MDA-MB-231) is an indirect method to assess PD-L1 endocytosis. PD-L1 molecules on the cell surface can be degraded by lysosomal and proteasome pathways, and small molecule inhibitors are added to induce PD-L1 endocytosis. After co-incubating small molecules with MDA-MB-231 cells for 24 hours, flow cytometry (Fluorescence-activated Cell Sorting, FACS) was used to detect the content of PD-L1 on the cell surface, which could indirectly reflect the endocytosis of PD-L1 induced by small molecules. Effect. Flow cytometry (FACS) was used to detect the effect of the compounds of the present invention on the expression level of PD-L1 in MDA-MR-231 cells.

Experimental Materials:

Phosphate buffered saline (DPBS), 1640 medium, penicillin-streptomycin, fetal bovine serum, non-essential amino acids, β-mercaptoethanol (2-ME), human interferon γ, LIVE/DEAD staining solution, staining solution (staining buffer), fixation buffer, 0.25% trypsin, EDTA, anti-human PD-L1 (Anti-human PD-L1), isotype control anti-human PD-L1 (Anti-human PD-L1 Isotype ).

1640 complete medium configuration: 439.5 ml of 1640 medium was added with 50 ml of fetal bovine serum, 5 ml of non-essential amino acids, 5 ml of penicillin-streptomycin and 0.5 ml of β liter mercaptoethanol, and mixed.

10mM EDTA configuration: add 1ml of 0.5M EDTA to 49ml of DPBS and mix.

Experimental steps:

1) MDA-MB-231 cell counting and plating: remove the culture flask, remove the medium and rinse once with DPBS. After washing, add 3 ml of 0.25% trypsin to the culture flask and place it in a 37°C incubator for 1.5 min. Remove the culture flask and add 9 ml of 1640 complete medium to stop the reaction, transfer the cells to a 50 ml centrifuge tube, and centrifuge at 1000 rpm at 37°C for 5 min. Add an appropriate volume of culture medium to resuspend the cells according to the number of cells, and count with a cell counter. The cell concentration was adjusted to 5 x 10 ⁵ cells/ml with the medium. Plating: A volume of 200 μL of cell suspension was added to each well of a 96-well plate, so that the number of cells in each well was 1×10 ⁵ . Incubate overnight in an incubator.

2) Drug incubation: prepare 100X compound diluent, and dilute the drug in a 5-fold gradient. Add 2 μL of each 100X compound solution to each well of cells. Incubate in a 37°C incubator for 24 hours. 3) PD-L1 cell staining and FACS detection: take out the culture plate and discard the supernatant medium. Wash once with 200 μL 1XPBS. Add 100 μL of EDTA (final concentration of 10 mM) at 37°C for 10 min. After centrifugation at 1500 rpm for 5 min, wash once with 200 μL of staining solution. Staining: Dilute anti-human PD-L1 (2 μL per well) and LIVE/DEAD staining solution (1:1000) in staining solution, add 50 μL to each well, and stain at 4°C for 30 min. Wash twice with 200 μL of staining solution. Fixation: Add 100 μL of fixative to each well, and fix at 4°C for 15 min. Wash twice with 200 μL of staining solution. Resuspend cells in 150 μL. FACS detection. Table 2 provides the effects of the compounds of the examples of the present invention on the expression level of PD-L1 in MDA-MR-231 cells.

Table 2 Test results of the effect of the compounds of the examples of the present invention on the expression level of PD-L1 in MDA-MR-231 cells

受试化合物test compound	IC ₅₀(nM) _IC50 (nM)
化合物1的甲酸盐Formate salt of compound 1	3.413.41
化合物2Compound 2	3.523.52
化合物3Compound 3	4.074.07
化合物8的甲酸盐Formate salt of compound 8	5.855.85
化合物12Compound 12	3.823.82
化合物15Compound 15	2.132.13

Experimental conclusion: The compound of the present invention has a significant inhibitory effect on the expression level of PD-L1 in MDA-MR-231 cells.

Experimental example 3: NFAT activity test

Experimental principle:

The engineered T cells express PD-1 molecule and T cell receptor (TCR) on the surface, which can activate the NFAT signaling pathway of T cells after co-culture with engineered antigen presenting cells (APC). The expression of PD-L1 molecules on APCs can effectively attenuate the NFAT signaling pathway in T cells; the use of PD-L1 inhibitors can effectively block the PD-1/PD-L1 regulatory mechanism, thereby reversing the weakened NFAT signaling pathway. After pretreatment with APC, the small molecule was co-cultured with T cells, and then the expression of luciferase was detected to indirectly reflect the activation of the NFAT pathway in T cells.

Experimental Materials:

PD-1/PD-L1 NFAT detection kit was purchased from BPS Biosciences. Birght-Glo reagent was purchased from Promega. Nivo Multilabel Analyzer (PerkinElmer).

experimental method:

The TCR Activitor/PD-L1 CHO cells with a growth confluence of 80% were plated into the plate at 35,000 cells per well and placed in a 37°C cell culture incubator overnight; the compounds to be tested were diluted 5-fold to the 8th Each concentration was diluted from 20 μM to 0.25 nM, and the DMSO concentration was 2%, and a double-well experiment was set up. Discard the TCR Activitor/PD-L1 CHO cell supernatant, add 50 μl of compound working solution to each well, and incubate at 37°C for 30 minutes; after the incubation, add 50 μL of PD-1/NFAT Reporter-Jurkat at a density of 4X10 ⁵ /ml to each well. The cell suspension was incubated at 37°C for 5 hours. After the incubation, 100 μL Bright-Glo was added to each well, and after mixing, the chemiluminescence signal was read using a Nivo multi-label analyzer.

data analysis:

Using the equation (sample-Min)/(Max-Min)×100% to convert the raw data into inhibition rate, the IC ₅₀ value can be obtained by curve fitting with four parameters (log(inhibitor) vs.response in GraphPad Prism --Variable slope mode). Table 3 provides the inhibitory activity of the compounds of the examples of the present invention on PD-1/PD-L1 binding.

Table 3 Test results of the inhibitory activity of the compounds of the examples of the present invention on the binding of PD-1/PD-L1

Experimental conclusion: The compound of the present invention can inhibit the interaction of PD-1/PD-L1 at the cellular level, thereby significantly activating the NFAT signaling pathway of T cells.

Experimental Example 4: Pharmacokinetic Testing

Objective: To study the pharmacokinetics of compounds in C57BL/6 mice

Experimental materials: C57BL/6 mice (male, 8 weeks old, body weight 25g-30g)

Experimental operation: The pharmacokinetic characteristics of rodents after intravenous injection (IV) and oral (PO) administration of the compound were tested according to the standard protocol. In the experiment, the candidate compound was formulated into a 1 mg/mL clear solution and administered to mice by a single intravenous injection and oral administration. Dosing. Intravenous and oral vehicles are 5% DMSO/5% polyethylene glycol 15-hydroxystearate (Solutol)/90% aqueous solution. This project uses four male C57BL/6 mice, and two mice are administered intravenously at a dose of 1 mg/kg. 24h plasma samples, the other two mice were given oral gavage at a dose of 10 mg/kg, and the plasma samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 h after administration. Collect whole blood samples within 24 hours, centrifuge at 3000g for 15 minutes, separate the supernatant to obtain plasma samples, add acetonitrile solution containing internal standard to precipitate proteins, thoroughly mix and centrifuge to take the supernatant for injection, and quantify by LC-MS/MS analysis method Analyze blood drug concentrations and calculate pharmacokinetic parameters, such as peak concentration (C _max ), clearance (CL), half-life (T _1/2 ), tissue distribution (V _dss ), area under the drug-time curve (AUC _{0- last} ), bioavailability (F), etc.

The pharmacokinetic-related parameters of the compounds of the examples of the present invention in mice are shown in the following table.

Table 4 Pharmacokinetic test results

Experimental conclusion: The compound of the present invention has good pharmacokinetic properties, including good oral bioavailability, oral exposure, half-life and clearance rate.

Experimental Example 5: Pharmacodynamic evaluation of the compound in the C57BL/6-hPDL1 mouse colorectal cancer MC38-hPDL1 subcutaneous transplantation model Experimental purpose: To evaluate the compound in the mouse colorectal cancer MC38-hPDL1 transplanted humanized mice Antitumor effects in C57BL/6-hPDL1.

Table 5 Experimental Design

Remarks: G: group; N: number of animals; p.o: oral administration; BID: twice a day.

Table 6 Experimental animals

种属species	小鼠mouse
品系strain	C57BL/6-hPDL1C57BL/6-hPDL1
级别level	SPF级SPF grade
周龄Zhou age	5.86～6.865.86～6.86
性别gender	雌性female

experimental method:

1. Tumor Cell Inoculation

Experimental cells: mouse colon cancer cells MC38-hPDL1 were recovered, and the recovery time was Pn+6. The MC38-hPDL1 cells in logarithmic growth phase were collected, and the culture medium was removed and washed twice with PBS before inoculation (before tumor-bearing, tumor-bearing The survival rates of MC38-hPDL1 cells were: 97.4% and 95.0%, respectively), inoculation volume: 1×10 ⁶ /100 μL/cell, inoculation location: right forelimb of mice.

2. Group administration

On the 7th day after inoculation, when the average tumor volume reached 85.23 mm ³ , the mice were randomly divided into 5 groups according to the tumor volume, with 8 mice in each group. The day of grouping was defined as D0 day, and the administration started on D0 day. The remaining mice were used for subsequent supplementary experiments.

3. Drug preparation

Dosing volume: adjusted according to mouse body weight (mice dosing volume = 10 μL/g × mouse body weight (g))

4. Experimental observation and data collection

Tumor size was observed on days 0, 2, 4, 6, 8, 11, 13, 15, 18, 20, 22, and 25 after the start of administration. The tumor volume was calculated as follows: tumor volume (mm ³ )=0.5×(longer diameter of tumor×shorter diameter of ^tumor2 ).

5. Experiment end point

At the end of the experiment, the following indicators were analyzed: 1) tumor volume change (TGItv); 2) average body weight change; TGITV (relative tumor inhibition rate) calculation formula:

TGItv(%)=[1-(meanTVtn-meanTVt0)/(meanTVvn-mean TVv0)]×100%

meanTVtn: the mean tumor volume of a given group when measured on day n

meanTVt0: the mean tumor volume of a given group when measured on day 0

meanTVvn: mean tumor volume of the solvent control group when measured on day n

mean TVv0: mean tumor volume of the solvent control group measured on day 0

Experimental results: The evaluation of the antitumor efficacy of the compounds of the present invention on the C57BL/6-hPDL1 mouse colorectal cancer MC38-hPDL1 subcutaneous transplantation model (calculated based on the tumor volume on the 25th day after administration) is shown in Table 7 below:

Table 7

The effects of the compounds of the present invention on the average body weight of mice in the colorectal cancer MC38-hPDL1 subcutaneous transplantation model of C57BL/6-hPDL1 mice are shown in Table 8 below:

Table 8

组别group	给药前(第0天)平均体重(克)Average body weight (g) before dosing (day 0)	给药第25天平均体重(克)Average body weight on day 25 of dosing (grams)
Vehicle(空白组)Vehicle (blank group)	19.219.2	20.020.0
化合物8的甲酸盐Formate salt of compound 8	19.019.0	19.919.9
化合物12Compound 12	19.219.2	19.319.3
化合物15Compound 15	19.519.5	20.020.0
化合物2Compound 2	19.019.0	18.818.8

Experimental conclusion: The compound of the present invention has excellent tumor-inhibiting effect on the colorectal cancer MC38-hPDL1 subcutaneous transplantation model of C57BL/6-hPDL1 mice, and the animal body weight does not decrease significantly during the administration process, and the tolerance is good.

Experimental Example 6: Toxicological study on repeated administration of compounds by oral gavage in Beagle dogs for 28 days

Experimental purpose: To evaluate the compound in rats, Beagle dogs were given different doses of the compound by oral gavage once a day for 4 consecutive weeks, and the drug was discontinued and recovered for 4 weeks, and the toxicity and its reversible degree or possible delayed toxicity were observed. reaction, explore its toxic reaction, and determine the toxic target organ or target tissue.

Table 9: Experimental Design

组别group	剂量mg/kgDosagemg/kg	给药容量ml/kgDosing volume ml/kg	理论浓度mg/mlTheoretical concentration mg/ml	给药频率Dosing frequency
溶媒对照vehicle control	00	00	00	每天一次Once a day
低剂量low dose	1515	55	33	每天一次Once a day
中剂量medium dose	5050	55	1010	每天一次Once a day
高剂量high dose	150150	55	3030	每天一次Once a day

Table 10: Experimental animals

种属species	犬dog
品系strain	Beagle犬Beagle dog
级别level	普通级Ordinary grade
周龄Zhou age	6～8月龄6 to 8 months old
性别gender	雌雄male and female

Experimental methods: The experiments were designed and carried out according to the current guidelines and pre-experimental results.

Experimental conclusion: the body weight of the compounds of the present invention did not decrease significantly during the administration process, and the tolerance was good; compared with the vehicle control group, gastrointestinal reactions, cough, salivation and total bilirubin (TBIL) were mainly seen. . Target organs were liver, lung, thymus, spleen, mesenteric lymph nodes, submandibular lymph nodes, and ileum. All the above changes can be seen to recover or recover in the recovery period, and the compounds of the present invention have good safety.

Claims

a compound of formula (I) or a pharmaceutically acceptable salt thereof,

in,

R 1 and R 2 are each independently selected from H, F, Cl, Br, I, CN and C 1-3 alkyl optionally substituted with 1 , 2 or 3 halogens;

R 3 is selected from H, CN, C 1-3 alkyl, C 1-3 alkoxy and C 1-3 alkylamino, the C 1-3 alkyl, C 1-3 alkoxy and C 1- 3 alkylamino groups are each independently optionally substituted with 1, 2 or 3 halogens;

R 4 and R 5 are each independently selected from H, C 1-6 alkyl, C 1-6 alkylamino, C 3-6 cycloalkyl and -C 1-3 alkyl-(3-6 membered heterocycloalkane base), the C 1-6 alkyl, C 1-6 alkylamino , C 3-6 cycloalkyl and -C 1-3 alkyl-(3-6 membered heterocycloalkyl) are independently optional replaced by 1, 2 or 3 Ra ;

Alternatively, R 4 and R 5 are connected to form a 3-8 membered heterocycloalkyl, and the 3-8 membered heterocycloalkyl is optionally substituted with 1, 2 or 3 R b ;

R 6 is selected from C 1-3 alkyl optionally substituted with 1, 2 or 3 R c ;

L is selected from -C 1-6 alkyl- optionally substituted with 1, 2 or 3 R d ;

X is selected from CH and N;

Y is selected from CH and N;

Z 1 is selected from a single bond and CH 2 ;

Z 2 is selected from CH and N;

Ra is selected from F, Cl, Br, I, OH, =O, NH 2 , C 1-3 alkylamino, C 1-3 alkoxy and C 1-3 alkyl, the C 1-3 alkylamino , C 1-3 alkoxy and C 1-3 alkyl are each independently optionally substituted with 1, 2 or 3 R;

R b is selected from CN, F, Cl, Br, I, OH, -C(=O)NH 2 , C 1-3 alkylamino, C 1-3 alkoxy and C 1-3 alkyl;

R c is selected from F, Cl, Br, I, CN, OH, =O and NH2 ;

R d is selected from F, Cl, Br, I, CN, OH, =O and NH2 ;

R is selected from F, Cl, Br, I, CN, OH, =O and NH2 ;

The 3-6 membered heterocycloalkyl and the 3-8 membered heterocycloalkyl each independently contain 1, 2 or 3 heteroatoms or heteroatomic groups independently selected from N, O, S and NH.
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 are each independently selected from H, F, Cl, Br, I, CN, CF 3 and CH 3 .
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 3 is selected from H, CN, CH 3 , -OCH 3 and
The CH 3 , -OCH 3 and
are each independently optionally substituted with 1, 2 or 3 halogens.
The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 3 is selected from H, CN, CH 3 , -OCH 3 ,
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 4 and R 5 are each independently selected from H, C 1-4 alkyl, C 1-3 alkylamino, cyclopropyl and
The C 1-4 alkyl, C 1-3 alkylamino, cyclopropyl and
are each independently optionally substituted with 1, 2 or 3 R a .
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R a is selected from F, Cl, Br, I, OH, =O, NH 2 , -NHCH 3 , -OCH 3 , -CH 2 OH and CH3 .
The compound according to claim 5 or 6 or a pharmaceutically acceptable salt thereof, wherein R 4 and R 5 are independently selected from H,
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 4 and R 5 are linked to form pyrrolidinyl, 8-azabicyclo[3.2.1]octyl, azetidinyl and 2-azaspiro[3.3]heptyl, the pyrrolidinyl, 8-azabicyclo[3.2.1]octyl, azetidinyl and 2-azaspiro[3.3]heptane The groups are each independently optionally substituted with 1, 2 or 3 R b .
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R b is selected from CN, OH, -C(=O) NH2 , -OCH3 and CH3 .
The compound of claim 8 or 9, or a pharmaceutically acceptable salt thereof, wherein R 4 and R 5 are linked to form
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R 6 is selected from CH 3 .
The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein L is selected from
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit
selected from
The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit
selected from
The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 12, wherein the compound is selected from

wherein, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , and Z 1 are as defined in claims 1 to 12,

R7 and R8 are selected from H, F, Cl, Br, I, CN, OH, =O and NH2 .
The compound or a pharmaceutically acceptable salt thereof according to claims 1 to 4, wherein the compound is selected from

wherein R 1 , R 2 , R 3 , R 6 , Z 1 , Z 2 , X and Y are as defined in claims 1 to 4,

Ring A is selected from 3-8 membered heterocycloalkyl,

R 9 and R 10 are each independently selected from H, CN, F, Cl, Br, I, OH, -C(=O)NH 2 , C 1-3 alkylamino, C 1-3 alkoxy and C 1 -3 alkyl.
A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from
The compound of claim 17, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from
Use of the compound according to any one of claims 1-18 or a pharmaceutically acceptable salt thereof in the preparation of a PD-1/PD-L1 inhibitor.
The use according to claim 19, wherein the PD-1/PD-L1 inhibitor is an antitumor drug.