WO2022083687A1 - Selenium heterocyclic compounds and application thereof - Google Patents

Abstract

A class of selenium heterocyclic compounds and an application thereof. Specifically disclosed are a compound represented by formula (II) and a pharmaceutically acceptable salt thereof.

Description

Selenium heterocyclic compounds and their applications

This invention claims priority

CN202011136978.2, application date: October 21, 2020;

CN202110242560.8, application date: March 4, 2021.

technical field

The present invention relates to selenium heterocyclic compounds and applications thereof. Specifically, it relates to compounds represented by formula (II) and pharmaceutically acceptable salts thereof.

Background technique

Inflammatory bowel disease (IBD) is an inflammatory bowel disease of unknown etiology, including Crohn's disease (CD) and ulcerative colitis (UC), characterized by inflammation and ulceration of the mucosal layers of the rectum and large intestine. Common symptoms include diarrhea, bloody stools, and abdominal pain. The clinical course is intermittent, alternating cycles of exacerbations and remissions, and patients with ulcerative colitis have an increased risk of colorectal cancer (Dennis et al. N Engl J Med, 2011, 365, 1713-1725). Excessive inflammatory responses in the gastrointestinal tract are mediated by inflammatory cytokines such as TNFα, IFN-γ, IL-1, IL-6, IL-12, IL-21 and IL-23, and are Cells of the adaptive immune system act, including on T and B lymphocytes, epithelial cells, macrophages and dendritic cells (Neurath, M.F. Nat. Rev. Immunol. 2014, 14, 329). The Janus kinase (JAK) family: JAK1, JAK2, JAK3, and Tyk2, are non-receptor tyrosine kinases that play key roles in the transduction of many of the above-mentioned cytokines. When cytokines bind to receptors, the associated JAK homo- or heterodimers are phosphorylated and activated, allowing subsequent recruitment, phosphorylation, and activation of signal transducer and activator of transcription (STAT) family transcription factors . Phosphorylated STATs (pSTATs) are transported to the nucleus and induce gene transcription of several chemokines, cytokines and proteases associated with the pathogenesis of IBD.

Genetic studies of IBD patients have found that polymorphisms of several proteins associated with the JAK/STAT pathway (eg, IL-23R, IL-12B, JAK2, Tyk2, and STAT3) are risk factors for the development of IBD. It provides an attractive target for the treatment of excessive inflammatory responses caused by IBD (Boland, B.S. et al., Gastroenterology clinics of North America 2014, 43, 603). Several JAK inhibitors, such as Tofacitinib and Filgotinib, are currently in clinical development for IBD. Tovatinib is approved in the United States for the treatment of rheumatoid arthritis (RA) and UC. Tovatinib is also in clinical development for CD, and further trials for this indication were discontinued due to the failure of patients with moderate to severe CD to achieve significant efficacy in a 4-week phase II clinical trial, despite biomarker-based There is conclusive evidence for the involvement of the target of the biological analysis, but it is unclear whether the efficacy of tovatinib is related to clinical study design, mechanistic differences between UC and CD, or dose-limiting systemic dose-limiting that prevents adequate exposure of the drug to intestinal tissue adverse events (AEs). Common systemic adverse events (AEs) in Phase 2 and 3 clinical trials of tovatinib in IBD included decreased hemoglobin, decreased absolute neutrophil count (ANC), increased total cholesterol (low-density and high-density lipids), and Infection (Sandborn, W.J. et al., N. Engl. J. Med. 2012, 367, 616). Such AEs are consistent with those observed with tovatinib in patients with rheumatoid arthritis and are consistent with the JAK2-dependent inhibition of EPO, TPO. According to the results of the tovatinib post-marketing safety trial, the use of tovatinib 10 mg twice daily was associated with an increased risk of blood clots and death, and the FDA issued a boxed warning for tovatinib.

There are several possible approaches to overcome systemic AEs caused by JAK inhibition. One such approach is the development of oral JAK1-selective inhibitors, such as filgotinib and Upadacitinib, which are currently being used in Phase 3 clinical trials in CD and UC. Despite the theoretical safety advantage of the JAK1-selective molecule, the recently approved 15 mg twice-daily dose of upadacitinib for rheumatoid arthritis also has a boxed warning from the FDA about the risk of thrombosis (Upadacitinib Instructions for Use) ). Another approach is to maximize intestinal tissue exposure to JAK inhibitors while avoiding possible systemic exposure. Systemic exposure to JAK inhibitors may have adverse systemic immunosuppressive effects due to the regulatory effects of the JAK/STAT pathway on the immune system. There is thus a need to provide new JAK inhibitors that have their effects at the focal site without significant systemic effects. Specifically, it has advantages for the treatment of gastrointestinal inflammatory diseases such as UC and CD.

SUMMARY OF THE INVENTION

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

in,

L ₁ is -(CH ₂ ) _m -, -C(=O)-, -S(=O) ₂ - or -CH ₂ -C(=O)-;

T ₁ is N or CH;

T ₂ is N or CH;

_T3 is N or CH;

R ₁ is H, CN, C _1-3 alkyl, -NH-C _1-3 alkyl, 4-6 membered heterocycloalkyl or 5-6 membered heteroaryl, wherein the C _1-3 alkyl , -NH-C _1-3 alkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl, each independently optionally substituted with 1, 2 or 3 R _a ;

_R2 is _R21 or

D ₁ is O, NH or CH ₂ ;

D ₂ is CH or N;

E ₁ and E ₂ are each independently a single bond or CH ₂ ;

R ₂₁ is H, CN, NH ₂ , C _1-3 alkyl, -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ , -C(=O)-NH-C _{1 -3} alkyl and -C(=O)-NH-C _3-6 cycloalkyl, wherein said C _1-3 alkyl, -NH-C _1-3 alkyl, -N(C _1-3 alkane base) ₂ , -C(=O)-NH-C _1-3 alkyl and -C(=O)-NH-C _3-6 cycloalkyl, respectively, independently optionally replaced by 1, 2 or 3 R _b replace;

R ₃ is H or C _1-3 alkyl;

R ₄ is H or C _1-3 alkyl;

q is 1 or 2;

m is 0, 1 or 2;

n is 1 or 2;

_Ra and _Rb are each independently F, Cl, Br, I, OH, _NH2 , CN or COOH;

Said "4-6-membered heterocycloalkyl" and "5-6-membered heteroaryl" each independently contain 1, 2 or 3 independently -O-, -NH-, -S-, -Se- or A heteroatom or heteroatom group of N.

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

in,

L ₁ is -(CH ₂ ) _m -, -C(=O)-, -S(=O) ₂ - or -CH ₂ -C(=O)-;

T ₁ is N or CH;

T ₂ is N or CH;

R ₁ is H, CN, C _1-3 alkyl, -NH-C _1-3 alkyl or 4-6 membered heterocycloalkyl, wherein the C _1-3 alkyl, -NH-C _1-3 Alkyl and 4-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R _a ;

_R2 is _R21 or

D ₁ is O or CH ₂ ;

R ₂₁ is H, C _1-3 alkyl, -NH-C _1-3 alkyl or -N(C _1-3 alkyl) ₂ , wherein the C _1-3 alkyl, -NH-C _{1- 3} alkyl and -N(C _1-3 alkyl) ₂ are optionally substituted by 1, 2 or 3 R _b ;

R ₃ is H or C _1-3 alkyl;

q is 1 or 2;

m is 0, 1 or 2;

_Ra and _Rb are each independently F, Cl, Br, I, OH, _NH2 , CN or COOH.

The "4-6 membered heterocycloalkyl" contains 1, 2 or 3 heteroatoms or heteroatomic groups that are independently -O-, -NH-, -S-, -Se- or N.

In some solutions of the present invention, the above R ₁ is H, CN, CH ₃ , -NH-CH ₃ ,

wherein the CH ₃ , -NH-CH ₃ ,

Each independently is optionally substituted with 1, 2 or 3 R _a , R _a and other variables as defined herein.

In some embodiments of the present invention, the above R ₁ is H, CN, CH ₃ , -NH-CH ₃ or

wherein the CH ₃ , -NH-CH ₃ and

Optionally substituted with 1, 2 or 3 _Ra , _Ra and other variables as defined herein.

In some solutions of the present invention, the above R ₁ is H, CN, CF ₃ , -NH-CH ₃ ,

Other variables are as defined in the present invention.

In some aspects of the present invention, the above R ₁ is H, CN, CF ₃ , -NH-CH ₃ or

Other variables are as defined in the present invention.

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

for

Other variables are as defined in the present invention.

In some solutions of the present invention, the above R ₂₁ is H, CN, NH ₂ , CH ₃ ,

wherein the CH ₃ ,

Each independently is optionally substituted with 1, 2 or 3 R _b , R _b and other variables as defined herein.

In some solutions of the present invention, the above R ₂₁ is H, CN, NH ₂ , CH ₃ ,

In some solutions of the present invention, the above R ₂ is H, CN, NH ₂ , CH ₃ ,

In some solutions of the present invention, the above R ₂ is H, CH ₃ ,

the CH ₃ ,

Optionally substituted with 1, 2 or 3 R _b , other variables are as defined in the present invention.

In some solutions of the present invention, the above R ₂ is H, CH ₃ ,

Other variables are as defined in the present invention.

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

In some embodiments of the present invention, the above R ₄ is H or CH ₃ , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above-mentioned L ₁ is -CH ₂ -, -(CH ₂ ) ₂ -, -C(=O)-, -S(=O) ₂ - or -CH ₂ -C(=O) -, other variables are as defined in the present invention.

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

for

Other variables are as defined in the present invention.

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

for

Other variables are as defined in the present invention.

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

for

Other variables are as defined in the present invention.

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

for

Other variables are as defined in the present invention.

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

for

Other variables are as defined in the present invention.

There are still some solutions of the present invention which are obtained by any combination of the above variables.

In some aspects of the present invention, the above-mentioned compound is

in,

n, R ₁ , R ₂ , R ₃ , L ₁ , T ₃ , T ₂ and q are as defined in the present invention.

In some aspects of the present invention, the above-mentioned compound is

in,

R ₁ , R ₂ , R ₃ , L ₁ , T ₂ and q are as defined in the present invention.

The present invention also provides a compound of the following formula or a pharmaceutically acceptable salt thereof,

The present invention provides the application of the above compounds or their pharmaceutically acceptable salts in the preparation of medicines for treating JAK-related diseases.

The present invention provides use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating pan-JAK-related diseases limited to the intestinal tract.

The present invention also provides a method for treating intestinal-limited pan-JAK-related diseases in a subject in need thereof, comprising providing the subject with an effective dose of a compound as defined in any of the above technical solutions or a pharmaceutically acceptable salts or pharmaceutical compositions.

In some technical solutions of the present invention, the above-mentioned pan-JAK-related disease limited to the intestine is inflammatory bowel disease.

technical effect

The compounds of the present invention show good inhibition in the in vitro activity test of two isoforms of kinases, JAK1 and JAK2. The compounds of the present invention exhibited good inhibitory properties in the in vitro activity test of cell (THP1 and HT29) functional assays. The compounds of the present invention show good drug exposure levels in the small intestine and colon of rats, and the compounds have high small intestine/plasma and colon/plasma ratios, showing good tissue selectivity. The compounds of the present invention show good drug exposure levels in the small intestine and colon of mice, and the small intestine/plasma and colon/plasma ratios of the compounds are high, showing good tissue selectivity. In the oxazolone (OXA)-induced mouse enteritis model, the compounds of the present invention can alleviate the weight loss induced by OXA, significantly improve the disease activity index (DAI) score and the end point colon weight-length ratio, and exhibit a good therapeutic effect.

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 the 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. 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, 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. "Optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances 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

For the group in this connection mode, 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 _1-3 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, "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 "4-6 membered heterocycloalkyl" by itself or in combination with other terms denotes a saturated cyclic group consisting of 4 to 6 ring atoms, respectively, of which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S, -Se-, and N, and the remainder are carbon atoms, where the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., 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 "4-6 membered heterocycloalkyl", a heteroatom may occupy the position of attachment of the heterocycloalkyl to the remainder of the molecule. The 4-6 membered heterocycloalkyl includes 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyl and the like. Examples of 4-6 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.

Unless otherwise specified, the terms "5-6 membered heteroaryl ring" and "5-6 membered heteroaryl" are used interchangeably in the present invention, and the term "5-6 membered heteroaryl" means from 5 to 6 ring atoms It is composed of a monocyclic group with a conjugated π electron system, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. Where the nitrogen atom is optionally quaternized, the nitrogen and sulfur heteroatoms may be optionally oxidized (ie, NO and S(O) _p , p is 1 or 2). A 5-6 membered heteroaryl group can be attached to the remainder of the molecule through a heteroatom or a carbon atom. The 5-6 membered heteroaryl groups include 5- and 6-membered heteroaryl groups. Examples of the 5-6 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrrolyl, etc.) azolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5- oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1, 2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl , 4-thiazolyl and 5-thiazolyl, etc.), furyl (including 2-furyl and 3-furyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2- -pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

Unless otherwise specified, _Cn-n+m or _Cn - _Cn+m includes any particular instance of n to n+ _m carbons, eg _C1-12 includes _C1 , _C2 , C3, C ₄ , C ₅ , C ₆ , C ₇ , C ₈ , C ₉ , C ₁₀ , C ₁₁ , and C ₁₂ , also including any one range from n to n+m, eg C _1-12 includes C _1-3 , C _1-6 , C _1-9 , C _3-6 , C _3-9 , C _3-12 , C _6-9 , C _6-12 , and C _9-12 , etc.; in the same way, n yuan to n +m-membered means that the number of atoms in the ring is from n to n+m, for example, 3-12-membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring , 10-membered ring, 11-membered ring, and 12-membered ring, also including any one range from n to n+m, for example, 3-12-membered ring includes 3-6 membered ring, 3-9 membered ring, 5-6 membered ring ring, 5-7 membered ring, 6-7 membered ring, 6-8 membered ring, and 6-10 membered ring, etc.

The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (eg, a nucleophilic substitution reaction). For example, representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as mesylate, tosylate, p-bromobenzenesulfonate, p-toluenesulfonic acid Esters, etc.; acyloxy, such as acetoxy, trifluoroacetoxy, and the like.

The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group" or "thiol protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: formyl; acyl groups, such as alkanoyl groups (eg, acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl groups, such as tert-butoxycarbonyl (Boc) ; Arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); Arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-di -(4'-Methoxyphenyl)methyl; silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS) and the like. The term "hydroxy protecting group" refers to a protecting group suitable for preventing hydroxyl side reactions. Representative hydroxy protecting groups include, but are not limited to: alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (eg acetyl); arylmethyl groups such as benzyl (Bn), p-methyl Oxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyl Dimethylsilyl (TBS) and the like.

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. The following abbreviations are used herein: aq stands for water; TFA stands for trifluoroacetic acid; ACN stands for acetonitrile; DMSO stands for dimethyl sulfoxide.

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 present invention has been described in detail herein, and specific embodiments thereof have also been disclosed. For those skilled in the art, 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. will be obvious.

Example 1

Step 1: Compound 1-1 (2 g, 11.56 mmol) was dissolved in dimethyl sulfoxide (10 mL) at 25 °C, 1-2 (2.88 g, 12.72 mmol) and N,N-diisopropyl were added Ethylamine (2.99 g, 23.12 mmol) was stirred at 100° C. for 16 hours, 200 mL of water was added to the reaction solution, and extracted with ethyl acetate (500 mL×3), the combined organic phases were then sequentially washed with 0.2 M aqueous hydrochloric acid ( 100 mL) and saturated brine (200 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 1-3. MS ESI calculated: _C18H23ClN4O2 [M ₊ H] _{+ 363} , found 363 ^.

Step 1': at 0 °C, compound 1-4a (10 g, 78.68 mmol) was dissolved in dichloromethane (100 mL), and N,N-diisopropylethylamine (12.20 g, 94.41 mmol, 16.45 mL) was added. ) and 1-4b ((15.74 g, 94.41 mmol, 16.71 mL), stirred at 25°C for 16 hours, 200 mL of water was added to the reaction solution, and extracted with ethyl acetate (500 mL×3), the combined organic phase was saturated with common salt Washed with water (200 mL), finally the organic phase was dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0～30/1) Compound 1-4c was obtained.

Step 2': Dissolve compound 1-4c (11g, 42.74mmol) in methanol (100mL), under nitrogen protection, add palladium carbon (0.1g, palladium content 10%), then replace with hydrogen three times, at 25 ℃ Stir for 16 hours, filter through celite and concentrate under reduced pressure to give crude 1-4. 1H NMR (400MHz, DMSO-d6) δ-0.05--0.03(s, 9H), 0.77-0.86(t, 2H), 1.95 -1.97(s, 3H), 3.47-3.53(t, 2H), 5.06-5.08(s, 1H), 5.08-5.11(s, 2H), 5.16(s, 2H).

Step 2: Compound 1-3 (2.5 g, 6.89 mmol) was dissolved in dioxane (30 mL), compound 1-4 (1.64 g, 7.23 mmol), cesium carbonate (4.49 g, 13.78 mmol) and [ (2-Di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino -1,1'-biphenyl)]palladium(II) methanesulfonate (312.28 mg, 344.49 μmol), was replaced with nitrogen three times and then heated to 100° C., and stirred for 16 hours under nitrogen protection. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentrated to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-5/1) to obtain compound 1-5. MS ESI calculated: C ₂₈ H ₄₃ N ₇ O ₃ Si[M+H] ⁺ 554, found 554. ¹ H NMR (400MHz, CD ₃ OD)δ-0.09--0.06(s, 9H), 0.81-0.88(t, 2H), 1.45-1.52(m, 11H), 1.76-1.85(m, 2H), 1.87 -2.01(m, 4H), 2.21-2.25(s, 3H), 3.49-3.55(t, 2H), 4.16-4.23(m, 2H), 4.33-4.48(m, 1H), 5.34(s, 2H) , 6.04-6.09 (m, 1H), 6.09-6.13 (m, 1H), 6.22-6.30 (s, 1H).

Step 3: Add selenium powder (4.39g, 54.17mmol) to ethanol (30mL) at 0-5°C, then slowly add sodium borohydride (2.38g, 62.91mmol), stir at room temperature until solid particles Completely disappeared, pyridine (8.57 g, 108.35 mmol) and compound 1-5 (3 g, 5.42 mmol) were added to the reaction solution, and the temperature was raised to 80 ° C and stirred for half an hour, and then slowly added 2M aqueous hydrochloric acid (32.50 mL), Continue stirring for half an hour, LC-MS showed that the raw materials were completely consumed, 50 mL of ammonium chloride aqueous solution was added to the reaction solution, and extracted with ethyl acetate (50 mL × 3), and the combined organic phases were washed with saturated brine (50 mL). , and finally the organic phase was dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0～1/1) to obtain compound 1-6 . MS ESI calculated: C ₂₈ H ₄₅ N ₇ O ₃ SeSi[M+H] ⁺ 636, found 636. ¹ H NMR (400MHz, CD ₃ OD) δ-0.02-0.01(s, 9H), 0.90-0.96(t, 2H), 1.52-1.55(m, 11H), 1.85-1.93(m, 2H), 1.96- 2.08(m, 4H), 2.24-2.30(s, 3H), 3.58-3.64(t, 2H), 4.22-4.31(m, 2H), 4.37-4.53(m, 1H), 5.36-5.43(s, 2H) ), 6.22-6.25(m, 1H), 6.32-6.35(s, 1H), 6.36-6.39(m, 1H).

Step 4: Compound 1-6 (1.4 g, 2.21 mmol) was dissolved in ethanol (15 mL) at 25 °C, 1-7 (204.07 mg, 2.21 mmol) was added, and the mixture was stirred at 80 °C for 1 hour. The reaction solution was directly concentrated under reduced pressure to obtain crude product 1-8.

Step 5: Compound 1-8 (1.4 g, 2.08 mmol) was dissolved in ethyl acetate (2 mL), ethyl acetate solution of hydrochloric acid (4 M, 14 mL) was added, and the mixture was stirred at 25° C. for 1 hour. The reaction solution was directly filtered to obtain the crude hydrochloride of compound 1-9 as the filter cake. MS ESI _calcd for _C20H25N7Se [ _M +H] ⁺ 444, found 444. ¹ H NMR (400MHz, CD ₃ OD) δ 1.93-2.05 (m, 2H), 2.20-2.42 (m, 9H), 2.56-2.62 (s, 3H), 4.13-4.26 (m, 3H), 5.98 ( s, 1H), 6.67 (s, 1H), 6.79 (s, 1H), 8.06-8.16 (s, 1H).

Step 6: The crude hydrochloride of 1-9 (0.85 g, 1.65 mmol) was dissolved in methanol (10 mL), N,N-diisopropylethylamine (639.52 mg, 4.95 mmol, 862 μL) was added, and the mixture was After stirring at 25°C for 10 minutes, compound 1-10 (0.290 g, 5.47 mmol, 363 μL) was added, and the mixture was stirred at 25°C for 16 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Luna C18 150*40mm*15μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 8%-38% , 11 minutes) to obtain the trifluoroacetate salt of compound 1-11. MS _ESI calculated for _C23H28N8Se [M+H] ⁺ 497, found 497, ^1H NMR (400MHz, _CD3OD ) δ 2.10-2.20 (m, 2H), _2.31-2.50 (m, 9H), 2.51-2.56(s, 3H), 3.10-3.17(t, 2H), 3.48-3.59(t, 2H), 4.17-4.31(m, 3H), 5.90-5.95(s, 1H), 6.68- 6.74 (m, 2H), 7.96 (m, 1H).

Example 2

Step 1: To a solution of 2-1 (10 mg, 81.23 μmol) in DMF (0.5 mL) at 25 °C was added N,N-diisopropylethylamine (21.0 mg, 162.46 μmol) and O-(7- Nitrobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (34.0mg, 89.35μmol), stirred at 25°C for 0.5h, and then added 1- A solution of 9's hydrochloride (41.86 mg, 81.23 μmol) and N,N-diisopropylethylamine (21.00 mg, 162.46 μmol, 28.30 μL) in DMF (0.5 mL) and stirred at 0-5 °C for 1 h . 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 24%-54% , 10 minutes) to obtain the trifluoroacetate salt of compound 2-2. MS ESI calculated for C ₂₆ H ₂₈ N ₈ OSe[M+H] ⁺ 549, found 549, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.88-2.01 (m, 2H), 2.07-2.16 (m, 2H), 2.17-2.27(m, 2H), 2.27-2.34(m, 2H), 2.34-2.38(s, 3H), 2.50-2.54(s, 3H), 4.25-4.36(m, 1H), 4.64- 4.70(m,1H),4.90-4.97(m,1H),5.84-5.89(s,1H),6.64-6.69(m,2H),7.51-7.57(m,1H),7.74-7.80(m,1H ), 7.94-8.01 (m, 2H), 8.59-8.67 (m, 1H).

Example 3

Step 1: The hydrochloride of compound 1-9 (50 mg, 97.02 μmol) was dissolved in methylpyrrolidone (1 mL) at 20 °C, triethylamine (49.09 mg, 485.10 μmol, 67.52 μL) was added, and the solution was heated at 20 °C. After stirring for 0.5 h, methylpyrrolidone (0.5 mL) was added to the reaction solution, the reaction solution was cooled to 0 °C, stirred for more than 0.5 h, 3-1 (21.03 mg, 116.42 μmol) was slowly added, the mixture was warmed to 20 °C and Stir for 3h. 20 mL of saturated ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phases were washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate and filtered. And concentrated under reduced pressure to obtain crude product. The crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 30%-60%, 10 minutes) to obtain compound 3 -2's trifluoroacetate. MS ESI calculated: C ₂₄ H ₂₉ N ₉ O ₂ SSe[M+H] ⁺ 588, found 588, ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 1.81-1.92 (m, 2H), 2.02- 2.10(m, 2H), 2.18-2.27(m, 4H), 2.35(s, 3H), 2.52(s, 3H), 3.66(m, 1H), 4.02(m, 2H), 4.09-4.18(m, 3H), 4.30(m, 2H), 5.87(s, 1H), 6.67(m, 2H), 7.97(s, 1H).

Example 4

Step 1: At 20°C, the hydrochloride salt of compound 1-9 (60 mg, 116.43 μmol) was dissolved in DMF (1 mL), potassium carbonate (32.18 mg, 232.86 μmol) and 4-1 (26.08 mg, 116.43 μmol) were added μmol), the mixture was stirred at 50° C. for 16 hours. 20 mL of water was added to the reaction solution, extracted with dichloromethane (10 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, Get crude. The crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 16%-46%, 10 minutes) to obtain compound 4 -2's trifluoroacetate. MS ESI calculated: C ₂₃ H ₂₈ F ₃ NSe[M+H] ⁺ 540, found 540, ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 2.08 (m, 2H), 2.31-2.38 (m, 5H), 2.39-2.49(m, 4H), 2.53(d, 3H), 2.81-2.93(m, 2H), 3.34-3.47(m, 2H), 4.14-4.33(m, 3H), 5.92(s, 1H), 6.64-6.79 (m, 2H), 7.96 (d, 1H).

Example 5

Step 1: Compound 5-1 (33.38 mg, 176.41 μmol) was dissolved in DMF (1 mL) at 20°C, hydroxybenzotriazole (47.67 mg, 352.81 μmol) and 1-(3-dimethylamino) were added propyl)-3-ethylcarbodiimide hydrochloride (67.63 mg, 352.81 μmol), and the mixture was stirred at 20° C. for 0.5 h. A solution of the hydrochloride salt of compound 1-9 (100 mg, 194.05 μmol) and diisopropylethylamine (68.40 mg, 529.22 μmol, 92.18 μL) in DMF (1 mL) was added to the mixture and stirred at 20° C. for 4 hours. 10 mL of water was added to the reaction solution, extracted with dichloromethane (20 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, Crude 5-2 was obtained. MS _ESI calculated: _C28H38N9O3Se [ _M +H] ⁺ ₆₁₅ , found 615.

Step 2: Compound 5-2 (150 mg, 244.45 μmol) was dissolved in dichloromethane (3 mL) at 20 °C, trifluoroacetic acid (1 mL) was added, and the mixture was stirred at 20 °C for 16 hours. The reaction solution was concentrated and spin-dried to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 13%-43%, 10 minutes) to obtain compound 5 -3's trifluoroacetate salt. MS ESI calculated: C ₂₃ H ₃₀ N ₈ OSe[M+H] ⁺ 515, found 515, ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 1.66-1.79 (m, 2H), 2.04-2.39 ( m, 10H), 2.52(d, 3H), 2.77(s, 3H), 4.00-4.18(m, 2H), 4.22-4.31(m, 1H), 4.31-4.38(m, 1H), 4.78(m, 1H), 5.88(s, 1H), 6.68(s, 2H), 7.97(d, 1H).

Example 6

Step 1: Compound 1-1 (200 mg, 1.16 mmol) and compound 6-1 (305.64 mg, 1.27 mmol) were dissolved in DMSO (2 mL) at 25 °C, and N,N-diisopropylethylamine ( 298.83 mg, 2.31 mmol), stirred at 100 °C for 2 hours, 10 mL of ethyl acetate was added to the reaction solution, washed with saturated brine (10 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure , to obtain crude 6-2. MS ESI calculated: _C19H25ClN4O2 [M ₊ H] _{+ 377} , found 377 ^.

Step 2: Compound 6-2 (300 mg, 796.01 μmol), compound 1-4 (217.19 mg, 955.21 mmol), cesium carbonate (518.71 mg, 1.59 mmol) and [(2-di-cyclohexylphosphino-3, 6-Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methane Palladium(II) sulfonate methanesulfonate (72.16 mg, 89.60 μmol) was dissolved in dioxane (3 mL), replaced with nitrogen three times, then heated to 100° C., and stirred under nitrogen protection for 16 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=20/1-5/1) to obtain compound 6-3. MS ESI calculated: C ₂₉ H ₄₅ N ₇ O ₃ Si[M+H] ⁺ 568, found 568.

Step 3: At 0-5°C, add selenium powder (285.23 mg, 3.52 mmol) to ethanol (4 mL), then slowly add sodium borohydride (159.91 mg, 4.23 mmol) in batches, and stir at room temperature until The solid particles disappeared completely. Pyridine (557.24 mg, 7.04 mmol) and compound 6-3 (200 mg, 352.24 μmol) were added to the reaction solution, and the temperature was raised to 80 °C and stirred for 0.5 hours, and then 2M aqueous hydrochloric acid (2.11 mL) was slowly added. , continue stirring for 0.5 hours, add 10 mL of water to the reaction solution, extract with ethyl acetate (10 mL×3), wash the combined organic phases with saturated brine (10 mL×2), and finally dry the organic phase with anhydrous sodium sulfate , filtered and concentrated under reduced pressure to obtain crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=10/0～1/1) to obtain compound 6-4. MS ESI calculated: C ₂₉ H ₄₇ N ₇ O ₃ SeSi[M+H] ⁺ 649, found 649.

Step 4: Compound 6-4 (140 mg, 159.69 μmol) was dissolved in ethanol (1 mL) at 25° C., compound 6-5 (17.73 mg, 196.62 μmol) was added, and the mixture was stirred at 80° C. for 1 hour. The reaction solution was directly concentrated under reduced pressure to obtain crude product 6-6. MS ESI calculated: C ₃₂ H ₄₉ N ₇ O ₃ SeSi[M+H] ⁺ 687, found 687.

Step 5: Compound 6-6 (110 mg, 160.16 μmol) was dissolved in ethyl acetate (1 mL), ethyl acetate solution of hydrochloric acid (4 M, 1.11 mL) was added, and the mixture was stirred at 25° C. for 12 hours. The reaction solution was directly filtered to obtain the crude hydrochloride of compound 6-7 as the filter cake. MS _{ESI calcd} for _C21H27N7Se [M+H] ⁺ 456, found 456.

Step 6: The crude hydrochloride of 6-7 (45 mg, 76.51 μmol) was dissolved in methanol (1 mL), N,N-diisopropylethylamine (49.44 mg, 382.53 μmol) was added, and the mixture was stirred at 25°C After 5 minutes, acrylonitrile (1-10) (12.18 mg, 229.52 μmol, 363 μL) was added, and the mixture was stirred at 25° C. for 16 hours. 10 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (20 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 15%-45% , 10 minutes) to obtain the trifluoroacetate salt of compound 6-8. MS ESI calculated for C ₂₄ H ₃₀ N ₈ Se[M+H] ⁺ 510, found 510, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.90-2.13 (m, 4H), 2.13-2.30 (m, 4H), 2.33(s, 1H), 2.36(s, 3H), 2.37-2.42(m, 1H), 2.49(s, 2H), 2.52(s, 3H), 3.10(t, J=7.25Hz, 2H ),3.79(t,J＝7.25Hz,2H),3.87(s,2H),4.69(tt,J＝11.76,6.07Hz,2H),5.94(s,1H),6.67(d,J＝1.13Hz , 1H), 6.69 (s, 1H), 7.94 (s, 1H).

Example 7

Step 1: The crude hydrochloride salt of compound 6-7 (45 mg, 76.51 μmol) was dissolved in methanol (1 mL), triethylamine (38.71 mg, 382.53 μmol) was added, and then compound 3-1 ( 16.58 mg, 91.81 μmol), stirred at 25°C for 16 hours. LC-MS showed that the starting material was completely consumed and the main peak was the product peak. 10 mL of aqueous solution was added to the reaction solution, extracted with ethyl acetate (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 the crude product, the crude product After preparative separation by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; B(ACN)%: 31%-61%, 10 minutes) to obtain The trifluoroacetate salt of compound 7-1. MS ESI calculated for C ₂₅ H ₃₁ N ₉ O ₂ SSe[M+H] ⁺ 601, found 601, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.87-1.99 (m, 5H), 2.00-2.08 ( m, 2H), 2.09-2.24(m, 1H), 2.29(d, 1H), 2.31(d, 1H), 2.36(s, 3H), 2.53(d, J＝0.88Hz, 3H), 3.65(m ,1H),3.99(m,2H),4.09-4.19(m,4H),4.60-4.74(m,1H),5.88(s,1H),6.66(s,2H),7.97(d,J=0.88 Hz, 1H).

Example 8

Step 1: Compound 8-1 (15 g, 78.13 mmol) was dissolved in dichloromethane (200 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (17.97 g) was added , 93.75 mmol) and 1-hydroxybenzotriazole (12.67 g, 93.75 mmol), were mixed and stirred at 20 °C for 1 hour, and then N,O-dimethylhydroxylamine hydrochloride (11.43 g, 117.19 mmol) and trimethylamine were added. Ethylamine (23.72 g, 234.38 mmol, 32.62 mL) was reacted at 20°C for 3 hours. The reaction solution was diluted with 400 mL of water and extracted with 450 mL of ethyl acetate (150 mL×3). The combined organic layers were washed with 200 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 3/1) to give compound 8-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.39 (s, 3H), 3.59 (s, 3H), 7.51 (s, 2H).

Step 2: Compound 8-2 (14.54 g, 61.85 mmol) was dissolved in tetrahydrofuran (200 mL), methylmagnesium bromide ether solution (3M, 24.74 mL) was added dropwise at 0 °C, and the reaction was carried out at 0 °C for 1 hour after the dropwise addition. . The reaction was quenched by adding 200 mL of saturated aqueous ammonium chloride solution, diluted with 200 mL of water, and extracted with 400 mL of ethyl acetate (200 mL×2). The combined organic layers were washed with 200 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude compound 8-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.62 (s, 3H), 7.68 (s, 2H).

Step 3: Compound 8-3 (11.65 g, 61.31 mmol) and ammonium chloride (327.94 mg, 6.13 mmol) were mixed, dichloromethane (200 mL) was added, and bromine (10.29 g, 64.37 mmol, 3.32 mL) was added dropwise , the reaction solution was reacted at 30-40 °C for 4 hours. 200 mL of saturated aqueous sodium bicarbonate solution was added, then diluted with 200 mL of water, extracted with dichloromethane 400 mL (200 mL×2), the combined organic layers were washed with brine 150 mL, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 8-4 . ¹ H NMR (400MHz, CDCl ₃ ) δ 4.36 (s, 2H), 7.73 (s, 2H)

Step 4: elemental selenium (2.89g, 35.67mmol, 2.83mL) was added to ethanol (30mL), sodium borohydride (1.83g, 48.37mmol) was added in batches at 0-5°C, and the reaction was performed at 20°C for 1 hour after the addition. Then 4-morpholinecarbonitrile (2 g, 17.84 mmol) was slowly added, then pyridine hydrochloride (8.24 g, 71.35 mmol) was slowly added, and the mixture was reacted at 20° C. for 16 hours. 50 mL of water was added to the reaction solution, extracted with dichloromethane (30 mL×3), the combined organic phases were washed with brine (30 mL×1), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to dryness in vacuo, toluene (20 mL) was added. ) was recrystallized and purified to obtain compound 8-6. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 3.52-3.59 (m, 4H), 3.60-3.97 (m, 4H), 7.93 (s, 2H).

Step 5: Compound 8-4 (278.52 mg, 1.04 mmol) was added to methanol (6 mL) and water (2 mL), sodium fluoride (21.74 mg, 517.85 μmol) and 8-6 (0.2 g, 1.04 mmol) were added, The reaction was carried out at 20°C for 1 hour. 20 mL of water was added, the solid was precipitated, filtered, and the filter cake was vacuum-dried to obtain compound 8-7. ¹ H NMR (400 MHz, CDCl ₃ ) δ 3.51-3.59 (m, 4H), 3.82-3.88 (m, 4H), 7.67 (s, 2H), 7.68 (s, 1H).

Step 6: Compounds 8-7 (1.71 g, 4.71 mmol), 1-2 (1.17 g, 5.18 mmol), [(2-di-cyclohexylphosphino-3,6-dimethoxy-2',4 ',6'-Triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)] palladium(II) methanesulfonate ( 213.45 mg, 235.50 μmol) and cesium carbonate (3.07 g, 9.42 mmol) were added to dioxane (35 mL) and reacted at 80° C. for 16 hours under nitrogen atmosphere. The reaction solution was diluted with 120 mL of water, extracted with 150 mL of ethyl acetate (50 mL×3), the combined organic layers were washed with brine 50 mL, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 3/1) to give compound 8-8. MS _ESI calculated: _{C24H32ClN5O3Se [ M} +H] ⁺ 554, found 554.

Step 7: Compounds 8-8 (680 mg, 1.12 mmol) and 8-9 (264.86 mg, 1.34 mmol) were added to toluene (15 mL) followed by [(2-di-cyclohexylphosphino-3,6-di Methoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]palladium methanesulfonate (II) Mesylate (101.44 mg, 111.91 μmol) and cesium carbonate (729.24 mg, 2.24 mmol) were reacted at 80° C. under nitrogen atmosphere for 16 hours. The reaction solution was diluted with 60 mL of water, and extracted with 60 mL (30 mL×2) of ethyl acetate. The combined organic layers were washed with 30 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 3/1) to give compound 8-10. MS _ESI calculated: _C33H46N8O5Se [M+H] ₊₇₁₅ , found ₇₁₅ .

Step 8: To compound 8-10 (160.67 mg, 225.11 μmol) was added ethyl acetate hydrochloride (4 M, 3 mL), and the reaction was carried out at 20° C. for 1.5 hours. The reaction solution was concentrated under reduced pressure to obtain compound 8-11. MS ESI calculated: C ₂₃ H ₃₀ N ₈ OSe [M+H]+515, found 515.

Step 9: Compound 8-11 (120 mg, 218.20 μmol) was dissolved in methanol (3 mL), diisopropylethylamine (141.00 mg, 1.09 mmol, 190.03 μL) and acrylonitrile (0.11 g, 2.07 mmol, 137.50 μL) were added μL), react at 20°C for 16 hours. The reaction solution was added with 30 mL of water, extracted with 40 mL (20 mL×2) of ethyl acetate, the combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography (column type: Phenomenex Synergi C18 150×25 mm×10 μm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; % acetonitrile: 16%-46%, 10 minutes) to obtain the trifluoroacetic acid salt of compound 8-12. MS ESI calculated: C ₂₆ H ₃₃ N ₉ OSe [M+H]+568, found 568. ¹ H NMR (400 MHz, CD ₃ OD) δ 2.12 (t, J=12.0 Hz, 2H), 2.27-2.38 (m, 5H), 2.38-2.50 (m, 4H), 3.10 (t, J=7.2 Hz) ,2H),3.42-3.62(m,6H),3.77-3.84(m,4H),4.15-4.23(m,1H),4.24(s,2H),5.83(d,J=0.4Hz,1H), 6.73 (d, J=1.2 Hz, 1H), 6.81 (d, J=1.2 Hz, 1H), 8.10 (s, 1H).

Example 9

Step 1: Compound 9-1 (15.36 mg, 105.84 μmol) was dissolved in DMF (0.5 mL) at 20°C, 1-hydroxybenzotriazole (28.60 mg, 211.69 μmol) and 1-(3- Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (40.58 mg, 211.69 μmol), and the mixture was stirred at 20° C. for 0.5 hr. A solution of the hydrochloride salt of compound 1-9 (60 mg, 116.43 μmol) and diisopropylethylamine (41.04 mg, 317.53 μmol, 55.31 μL) in DMF (0.5 mL) was added to the mixture, which was stirred at 20° C. for 16 hours . LC-MS showed that the starting material was consumed and the main peak was the product peak. 20 mL of water was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, Get crude. The crude product was separated by high performance liquid chromatography (column: Waters Xbridge 150*25mm*5μm; mobile phase: [water (10mM NH ₄ HCO ₃ )-ACN]; ACN%: 23%-53%, 8 minutes) to obtain the compound 9-2. MS ESI calculated: C ₂₆ H ₃₄ N ₈ O ₂ Se[M+H] ⁺ 571, found 571, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.24-1.41 (m, 1H), 1.50-1.71 (m,2H),1.87-2.38(m,11H),2.47(s,3H),2.54(m,4H),3.23(s,2H),3.68-3.77(m,4H),4.56(m,2H) ), 4.69 (m, 1H), 6.19-6.43 (m, 2H), 6.55 (m, 1H), 7.61-7.81 (m, 1H).

Example 10

Step 1: Compound 1-6 (80 mg, 126.03 μmol) was dissolved in ethanol (1 mL) at 20 °C, 10-1 (13.43 mg, 126.03 μmol, 12.79 μL) was added, and the mixture was stirred at 80 °C for 7 hours . The reaction solution was concentrated under reduced pressure to obtain crude product 10-2. MS _ESI calculated: _{C32H49N7O3SeSi} [ _M +H] ⁺ ₆₈₈ , found 688.

Step 2: Compound 10-2 (90 mg, 131.04 μmol) was dissolved in methanol (0.5 mL) at 20 °C, hydrochloric acid/ethyl acetate (4 M, 2 mL) was added, and the mixture was stirred at 20 °C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain crude 10-3 hydrochloride. MS ESI calculated: C ₂₁ H ₂₇ N ₇ Se[M+H] ⁺ 458, found 458.

Step 3: Compound 10-3 hydrochloride (80 mg, 162.30 μmol, HCl) was dissolved in methanol (1 mL) at 20°C, diisopropylethylamine (62.93 mg, 486.91 μmol, 84.81 μL) was added and Acrylonitrile (1-10) (120 mg, 2.26 mmol, 150.00 μL), and the mixture was stirred at 20° C. for 2 hours. 20 mL of water was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, Get crude. The crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 15%-45%, 10 minutes) to obtain compound 10 -4's trifluoroacetate salt. MS ESI calculated: C ₂₄ H ₃₀ N ₈ O ₈ Se[M+H] ⁺ 511, found 511, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.29 (m, 1H), 2.09 (m, 2H ), 2.29-2.49(m, 14H), 2.55(s, 3H), 3.09(t, 2H), 3.44-3.55(m, 2H), 4.24(m, 3H), 5.90(s, 1H), 6.63( m, 2H).

Example 11

Step 1: Elemental selenium (4.62g, 57.07mmol) was added to ethanol (40mL), sodium borohydride (2.19g, 57.89mmol) was added in batches at 0-5°C, reacted at 20°C for 1 hour after the addition, and then slowly added Dimethylcyanamide (11-1) (2 g, 28.53 mmol), then pyridine hydrochloride (13.19 g, 114.13 mmol) was slowly added, and the mixture was reacted at 20° C. for 16 hours. 100 mL of water was added to the reaction solution, extracted with dichloromethane:methanol=10:1 (50 mL×4), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to dryness in vacuo to obtain compound 11-2. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 3.12-3.45 (m, 6H), 7.62 (d, J=2.4Hz, 2H)

Step 2: Compound 8-4 (1 g, 3.72 mmol) was added to methanol (30 mL), compound 11-2 (561.76 mg, 3.72 mmol) was added, and the mixture was reacted at 20° C. for 1 hour. The pH of the system was adjusted to 7 by adding ammonia water, concentrated to dryness, and purified by silica gel column (petroleum ether:ethyl acetate=10:1) to obtain compound 11-3. ¹ H NMR (400MHz, CDCl ₃ ) δ 3.17(s, 6H), 7.58(s, 1H), 7.69(s, 2H)

Step 3: Compound 11-3 (0.8 g, 2.49 mmol), 1-2 (563.91 mg, 2.49 mmol), 2,2-bis(diphenylphosphino)-1,1-binaphthyl (155.15 mg, 249.17 μmol), palladium acetate (55.94 mg, 249.17 μmol) and cesium carbonate (1.62 g, 4.98 mmol) were added to dioxane (20 mL) and reacted at 90° C. for 5 hours under nitrogen atmosphere. The reaction solution was diluted with 100 mL of water, extracted with 100 mL of ethyl acetate (50 mL×2), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 20/1 to 5/1) to give compound 11-4. MS _ESI calculated: _{C22H30ClN5O2Se} [M ₊ H] ⁺ ₅₁₂ , found 512.

Step 4: Compound 11-4 (300 mg, 587.18 μmol), 8-9 (127.39 mg, 645.90 μmol) were added to dioxane (10 mL), followed by [(2-di-cyclohexylphosphino-3, 6-Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methane Palladium(II) sulfonate mesylate (53.23 mg, 58.72 μmol) and cesium carbonate (382.63 mg, 1.17 mmol) were reacted at 90° C. for 5 hours under nitrogen atmosphere. The reaction solution was diluted with 50 mL of water, and extracted with 40 mL (20 mL×2) of ethyl acetate. The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 3/1) purification to give compound 11-5. MS _ESI calculated: _C31H44N8O4Se [M ₊ H] ⁺ ₆₇₃ , found 673.

Step 5: Compound 11-5 (150 mg, 223.32 μmol) was dissolved in ethyl acetate (3 mL), ethyl acetate hydrochloride (4 M, 3 mL) was added, and the reaction was carried out at 20° C. for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 11-6. MS _ESI calculated: _C21H28N8Se [M+H] ⁺ ₄₇₃ , found 473.

Step 6: Compound 11-6 (150 mg, 295.32 μmol) was dissolved in methanol (5 mL), diisopropylethylamine (190.84 mg, 1.48 mmol, 257.20 μL) and acrylonitrile (0.1 g, 1.88 mmol, 125.00 were added) μL), react at 20°C for 16 hours. The reaction solution was added with 30 mL of water, extracted with 40 mL (20 mL×2) of ethyl acetate, the combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography (column type: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; % acetonitrile: 15%-45%, 10 minutes) to obtain the trifluoroacetate salt of compound 11-7. MS _ESI calculated: _C21H28N8Se [M+H] ⁺ ₄₇₃ , found 473. ¹ H NMR(400MHz,MeOD)δ2.13(m,2H),2.30-2.38(m,5H),2.38-2.52(m,4H),3.11(t,J=7.2Hz,2H),3.16(s ,6H),3.51(s,2H),4.12-4.23(m,1H),4.25(s,2H),5.83(s,1H),6.72(s,1H),6.81(d,J=0.8Hz, 1H),7.96(s,1H)

Example 12

Step 1: Compound 12-1 (3 g, 73.08 mmol, 3.85 mL) and selenium powder (12.84 g, 146.16 mmol) were added to DMF (30 mL) and water (3 mL), replaced 3 times with carbon monoxide, charged with carbon monoxide, kept The pressure was 50 psi, and the reaction was carried out at 80°C for 16 hours. After the reaction was completed, the oil was pumped to dryness, and purified by silica gel column (petroleum ether:ethyl acetate=1:2) to obtain compound 12-2. ¹ H NMR (400MHz, DMSO-d ₆ )δ2.39(s,3H),9.72-10.09(m,1H),10.23(s,1H)

Step 2: Compound 8-4 (3 g, 12.16 mmol) was added to methanol (120 mL), compound 12-2 (1.80 g, 12.27 mmol) was added, and the reaction was carried out at 20° C. for 2 hours. The pH of the system was adjusted to 7 by adding ammonia water, concentrated to dryness, and purified by silica gel column (petroleum ether:ethyl acetate=5:1) to obtain compound 12-3. ¹ H NMR (400MHz, CDCl ₃ ) δ 2.82(s, 3H), 7.75(s, 2H), 8.30(s, 1H)

Step 3: Compound 12-3 (0.5 g, 1.71 mmol), 1-2 (387.49 mg, 1.71 mmol), 2,2-bis(diphenylphosphino)-1,1-binaphthyl (106.61 mg, 171.22 μmol), palladium acetate (38.44 mg, 171.22 μmol) and cesium carbonate (1.12 g, 3.42 mmol) were added to dioxane (10 mL) and reacted at 90° C. for 5 hours under nitrogen atmosphere. The reaction solution was diluted with 100 mL of water, extracted with 100 mL of ethyl acetate (50 mL×2), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 20/1 to 5/1) to give compound 12-4. MS ESI calculated: _{C21H27ClN4O2Se} [M ₊ H] _{+ 483} , found 483 ^.

Step 4: Compounds 12-4 (590 mg, 1.22 mmol) and 8-9 (241.49 mg, 1.22 mmol) were added to dioxane (15 mL) followed by [(2-di-cyclohexylphosphino-3, 6-Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methane Palladium(II) sulfonate methanesulfonate (120.99 mg, 122.44 μmol) and cesium carbonate (797.85 mg, 2.45 mmol) were reacted at 90° C. for 5 hours under nitrogen atmosphere. The reaction solution was diluted with 50 mL of water, and extracted with 40 mL (20 mL×2) of ethyl acetate. The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 3/1) to obtain compound 12-5. MS _ESI calculated: _C30H41N7O4Se [M ₊ H] ⁺ ₆₄₄ , found 644.

Step 5: Compound 12-5 (150 mg, 210.07 μmol) was dissolved in ethyl acetate (3 mL), ethyl acetate hydrochloride (4 M, 2.70 mL) was added, and the reaction was carried out at 20° C. for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 12-6. MS _ESI calculated: _C20H25N7Se [ _M +H] ⁺ 444, found 444.

Step 6: Compound 12-6 (100 mg, 226.03 μmol) was dissolved in methanol (3 mL), diisopropylethylamine (146.06 mg, 1.13 mmol, 196.85 μL) and acrylonitrile (0.12 g, 2.26 mmol, 150.00 were added) μL), react at 15°C for 2 hours. The reaction solution was added with 30 mL of water, extracted with 40 mL (20 mL×2) of ethyl acetate, the combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography (column type: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; % acetonitrile: 13%-43%, 10 minutes) to obtain the trifluoroacetate salt of compound 12-7. MS ESI calculated: _C23H28N8Se [M+H] ₊₄₉₇ , found ₄₉₇ . ¹ H NMR (400MHz, CD ₃ OD) δ 2.13(t, J=12.4Hz, 2H), 2.35(s, 3H), 2.35-2.50(m, 6H), 2.81(s, 3H), 3.10(t , J=7.2Hz, 2H), 3.51(s, 2H), 4.14-4.31(m, 3H), 5.85(s, 1H), 6.77(d, J=1.2Hz, 1H), 6.84(d, J= 1.2Hz, 1H), 8.77(s, 1H).

Example 13

Step 1: Dissolve the crude hydrochloride of 6-7 (55 mg, 96.41 μmol) in DMF (1 mL), add N,N-diisopropylethylamine (37.38 mg, 289.24 μmol), and stir at 0°C 0.5 h, then compound 5-1 (16.58 mg, 91.81 μmol) and 1-hydroxybenzotriazole (23.69 mg, 175.29 μmol) and 1-(3-dimethylaminopropyl)-3- were added at 0 °C A solution of ethylcarbodiimide hydrochloride (36.96 mg, 192.82 μmol) in DMF (1 mL) was stirred at 25° C. for 15.5 hours. LC-MS showed that the starting material was almost completely consumed. The reaction solution was diluted with 10 mL of ethyl acetate, washed with saturated brine (10 mL×2), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was separated by preparative chromatography (SiO ₂ , PE:EA=0:1) to obtain compound 13-1. MS ESI _calcd for _C29H40N8O3Se [ _M +H] ⁺ ₆₂₈ , found 628.

Step 2: Dissolve 13-1 (20 mg, 31.87 μmol) in dichloromethane (1 mL), add trifluoroacetic acid (616 mg, 5.4 mmol), stir at 0°C for 16 hours, LC-MS shows that the starting material is basically consumed The main peak is the product peak. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was prepared and separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10 μm; mobile phase: [water (0.1% TFA)-ACN]; B(ACN)%: 17 %-47%, 10 min) to give compound 13-2 as the trifluoroacetate salt. MS _ESI calculated for _C24H32N8OSe [ _M +H] ⁺ 528, found 528, ^1H NMR (400MHz, _CD3OD ) δ 1.29 (m, 2H), 1.71-1.81 (m, 1H) ,1.86(m,1H),1.91(m,1H),1.94(m,2H),1.98-2.04(m,2H),2.27(m,1H),2.35(s,3H),2.37-2.46(m ,2H),2.53(s,3H),2.77(s,3H),4.11(s,2H), 4.69-4.76(m,1H),5.90(s,1H),6.67(s,1H),6.68( s, 1H), 7.97 (s, 1H).

Example 14

Step 1: The crude hydrochloride of 6-7 (55 mg, 96.41 μmol) was dissolved in DMF (2 mL), potassium carbonate (53.30 mg, 385.65 μmol) and compound 4-1 (43.19 mg, 192.82 μmol) were added at 120 Stir at °C for 3 hours. The reaction solution was diluted with 10 mL of water and extracted with ethyl acetate (10 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 40%-70%, 8 minutes) then (column: Phenomenex Synergi C18 150*25mm *10 μm; mobile phase: [water (0.1% TFA)-ACN]; B(ACN) %: 15%-45%, 10 minutes) to give the trifluoroacetate salt of compound 14-1. MS ESI calculated for C ₂₄ H ₃₀ F ₃ N ₇ Se[M+H] ⁺ 553, found 553, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.95-2.01 (m, 1H), 2.03-2.12 ( m, 1H), 2.12-2.28(m, 5H), 2.29-2.35(m, 2H), 2.35(s, 3H), 2.45-2.51(m, 1H), 2.52(s, 3H), 2.85(m, 2H),3.65-3.72(m,2H),3.87(s,2H),4.65-4.80(m,1H),5.97(s,1H),6.66(s,1H),6.69(s,1H),7.94 (s, 1H).

Example 15

Step 1: Compound 15-1 (2 g, 13.23 mmol) and diisopropylethylamine (2.56 g, 19.84 mmol) were added to tetrahydrofuran (30 mL), nitrile bromide (1.78 g, 16.80 mmol) was added at 0°C, 1.24 mL), reacted at 20°C for 16 hours. The reaction solution was diluted with 100 mL of water, extracted with 120 mL of ethyl acetate (40 mL×3), the combined organic layers were washed with 50 mL of brine, and dried over sodium sulfate to obtain compound 15-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ 2.68 (s, 3H), 3.74 (s, 3H), 4.01 (s, 2H), 6.84 (d, J=8.8 Hz, 2H), 7.14-7.23 (m, 2H).

Step 2: Elemental selenium (2.10g, 25.99mmol) was added to ethanol (50mL), sodium borohydride (983.24mg, 25.99mmol) was added in batches at 0-5°C, reacted at 20°C for 1 hour after the addition, and then slowly added Compound 15-2 (2.29 g, 13.00 mmol) was then slowly added with pyridine hydrochloride (6.01 g, 51.98 mmol), and the mixture was reacted at 20° C. for 16 hours. 100 mL of water was added to the reaction solution, extracted with dichloromethane:methanol=10:1 (50 mL×3), dried over anhydrous Na2SO4, filtered and concentrated to dryness in vacuo, purified by silica gel column (petroleum ether:ethyl acetate=10: 1 to 2:1) to give compound 15-3. ¹ H NMR (400MHz, DMSO-d ₆ ) δ 2.58-3.04(m, 3H), 3.49-3.84(m, 3H), 4.50-5.49(m, 2H), 6.67-7.00(m, 2H), 7.03 -7.43(m, 2H), 7.43-8.17(m, 2H).

Step 3: Compound 8-4 (0.6 g, 2.23 mmol, ) was added to methanol (40 mL), compound 15-3 (631.21 mg, 2.45 mmol) was added, and the reaction was carried out at 20° C. for 1 hour. After completion of the reaction, filter and vacuum dry the filter cake to obtain compound 15-4. MS ESI calculated: _{C17H15Cl2N3OSe} [ _{M +} H] ⁺ ₄₂₈ , found 428.

Step 4: Compound 15-4 (560 mg, 1.31 mmol), 1-2 (296.68 mg, 1.31 mmol), 2,2-bis(diphenylphosphino)-1,1-binaphthyl (29.43 mg, 131.09 μmol ), palladium acetate (29.43 mg, 131.09 μmol) and cesium carbonate (854.24 mg, 2.62 mmol) were added to dioxane (15 mL) and reacted at 90° C. for 16 hours under nitrogen atmosphere. The reaction solution was diluted with 100 mL of water, extracted with 100 mL of ethyl acetate (50 mL×2), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 20/1 to 5/1) to give compound 15-5. MS _ESI calculated: _{C29H36ClN5O3Se} [ _M +H] ⁺ ₆₁₈ , found 618.

Step 5: Compound 15-5 (440 mg, 648.91 μmol), 8-9 (127.99 mg, 648.91 μmol) were added to dioxane (15 mL), followed by [(2-di-cyclohexylphosphino-3, 6-Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methane Palladium (II) sulfonate methanesulfonate (58.82 mg, 64.89 μmol), cesium carbonate (422.85 mg, 1.30 mmol), reacted at 90° C. for 5 hours under nitrogen atmosphere. The reaction solution was diluted with 50 mL of water, and extracted with 40 mL (20 mL×2) of ethyl acetate. The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 3/1) to give compound 15-6. MS _ESI calculated: _C38H50N8O5Se [M+ _H ] ⁺ ₇₇₉ , found 779.

Step 6: Compound 15-6 (150 mg, 223.32 μmol) was dissolved in trifluoroacetic acid (7.70 g, 67.53 mmol, 5 mL) and reacted at 70° C. for 20 hours. The reaction solution was concentrated under reduced pressure to obtain compound 15-7. MS ESI calculated: _C20H26N8Se [ _M +H] ₊₄₅₉ , found 459.

Step 7: Compound 15-7 (90 mg, 157.49 μmol) was dissolved in methanol (3 mL), diisopropylethylamine (101.77 mg, 787.46 μmol, 137.16 μL) and acrylonitrile (0.15 g, 2.07 mmol, 137.50 μL) were added μL), react at 15°C for 2 hours. The reaction solution was added with 30 mL of water, extracted with 40 mL (20 mL×2) of ethyl acetate, the combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography (column type: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; % acetonitrile: 10%-40%, 10 minutes) to obtain the trifluoroacetate salt of compound 15-8. MS _ESI calculated: _C23H29N9Se [M+H] ⁺ ₅₁₂ , found 512. ¹ H NMR (400MHz, CD ₃ OD) δ 7.85(s, 1H), 6.69(s, 1H), 6.64(d, J=0.8Hz, 1H), 5.85(s, 1H), 4.24(s, 2H) ), 4.14-4.22(m, 1H), 3.51(s, 2H), 3.10(t, J=7.2Hz, 2H), 3.03(s, 3H), 2.38-2.48(m, 4H), 2.28-2.38( m, 5H), 2.12 (t, J=12.4Hz, 2H).

Example 16

Step 1: Compound 16-1 (11.88 mg, 139.71 μmol) was dissolved in DMF (0.5 mL) at 20°C, O-(7-azabenzotriazole-1-yl)-N was added, N,N,N-tetramethylurea hexafluorophosphonium salt (88.54 mg, 232.86 μmol, 34.62 μL), and the mixture was stirred at 20° C. for 0.5 hr. A solution of Compound 1-9 hydrochloride (60 mg, 116.43 μmol) and diisopropylethylamine (45.14 mg, 349.28 μmol, 60.84 μL) in DMF (0.5 mL) was added to the mixture and stirred at 20° C. for 16 hours. 10 mL of water was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, Get crude. The crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 23%-53%, 10 minutes) to obtain compound 16 -2's trifluoroacetate. MS ESI calculated: C ₂₃ H ₂₆ N ₈ OSe [M+H] ⁺ 511, found 511, ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.73-1.84 (m, 2H), 1.85-1.98 (m, 2H), 1.98-2.06(m, 2H), 2.13-2.29(m, 4H), 2.34(s, 3H), 2.55(s, 3H), 3.45-3.56(m, 2H), 4.01(m, 1H) ,4.26-4.33(m,1H),4.76-4.84(m,1H),5.76(s,1H),6.47(s,1H),6.64(s,1H),7.63(s,1H),8.27(s , 1H).

Example 17

Step 1: Compound 1-3 (2 g, 5.51 mmol) was dissolved in tetrahydrofuran (20 mL) at 0°C, sodium hydrogen (330.71 mg, 8.27 mmol, 60% purity) was added, and the mixture was stirred at 0°C for 0.5 hours. Iodomethane (0.17 g, 8.27 mmol, 514.70 μL) was added to the reaction solution, and the mixture was stirred at 20° C. for 1 hour. 30 mL of saturated aqueous sodium bicarbonate solution was added to the reaction solution, extracted with ethyl acetate (30 mL×3), the combined organic phases were washed with saturated brine (5 mL×3), and finally the organic phase was dried with anhydrous sodium sulfate, filtered and reduced Concentrate under pressure to obtain a crude product, which is purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=50/1-20/1) to obtain compound 17-1. MS ESI calculated: _C19H25ClN4O2 [M ₊ H] _{+ 377} , found 377 ^.

Step 2: Compound 17-1 (1.86 g, 4.94 mmol) was dissolved in dioxane (20 mL), compound 1-4 (1.18 g, 5.18 mmol), cesium carbonate (3.22 g, 9.87 mmol) and [ (2-Di-cyclohexylphosphino-3,6-dimethoxy-2′,4′,6′-triisopropyl-1,1′-biphenyl)-2-(2′-amino -1,1'-biphenyl)]palladium(II) methanesulfonate methanesulfonate (223.69 mg, 246.76 μmol), replaced with nitrogen three times and then heated to 100° C., and stirred for 16 hours under nitrogen protection. 30 mL of water was added to the reaction solution, extracted with ethyl acetate (40 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure , the crude product was obtained, and the crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=10/1～5/1) to obtain compound 17-2. MS ESI calculated: C ₂₉ H ₄₅ N ₇ O ₃ Si[M+H] ⁺ 568, found 568.

Step 3: Add selenium powder (885.68mg, 10.94mmol) to ethanol (20mL) at 0-5°C, then add sodium borohydride (413.75mg, 10.94mmol) slowly, stir at room temperature for half an hour , until the solid particles completely disappeared, pyridine hydrochloride (1.69g, 14.58mmol) and compound 17-2 (2.07g, 3.65mmol) were added to the reaction solution, and the temperature was raised to 80 ° C and stirred for 2 hours, and added to the reaction solution. 40 mL of water, extracted with ethyl acetate (30 mL×3), the combined organic phases were washed with saturated brine (15 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was passed through the column Chromatography (SiO ₂ , petroleum ether: ethyl acetate=8/1～2/1) was purified to obtain compound 17-3. MS ESI calculated value: C ₂₉ H ₄₇ N ₇ O ₃ SeSi[M+H] ⁺ 650, found 650.

Step 4: Compound 17-3 (300 mg, 462.41 μmol) was dissolved in ethanol (3 mL) at 25° C., 6-5 (300 mg, 3.24 mmol) was added, and the mixture was stirred at 80° C. for 2 hours. The reaction solution was directly concentrated under reduced pressure to obtain crude product 17-4. MS ESI _calcd for _{C32H49N7O3SeSi} [ _M +H] ⁺ ₆₈₈ , found 688.

Step 5: Compound 17-4 (200 mg, 359.35 μmol) was dissolved in methanol (0.5 mL), ethyl acetate solution of hydrochloric acid (4 M, 3 mL) was added, and the mixture was stirred at 20° C. for 16 hours. The reaction solution was directly spin-dried to obtain the crude hydrochloride of compound 17-5. MS _{ESI calcd} for _C21H27N7Se [M+H] ⁺ 458, found 458.

Step 6: Dissolve the crude hydrochloride of 17-5 (160 mg, 324.61 μmol) in methanol (1 mL), add N,N-diisopropylethylamine (125.86 mg, 973.82 μmol, 169.62 μL), then add Acrylonitrile (1-10) (120 mg, 2.26 mmol, 150.00 μL) was stirred at 20° C. for 2 hours. 20 mL of water was added to the reaction solution, extracted with dichloromethane (20 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure The crude product was obtained, and the crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 16%-46%, 10 minutes) The trifluoroacetate salt of compound 17-6 was obtained. MS ESI calculated for C ₂₄ H ₃₀ N ₈ Se[M+H] ⁺ 511, found 511, ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.26 (s, 1H), 1.91-2.00 (m, 2H), 2.30-2.40(m, 7H), 2.45-2.62(m, 6H), 2.97(s, 3H), 3.14(m, 2H), 3.30(m, 2H), 4.16(m, 2H), 4.86-5.04( m, 1H), 6.07(s, 1H), 6.58(s, 1H), 6.94(s, 1H), 7.63(s, 1H), 10.60-10.77(m, 1H).

Example 18

Step 1: Compound 18-1 (0.4 g, 2.3 mmol) was dissolved in dimethyl sulfoxide (2 mL) at 25 °C, 1-2 (520.30 mg, 2.30 mmol) and N,N-diiso were added Propylethylamine (594.3 mg, 4.60 mmol, 800.89 μL) was stirred at 25°C for 1 hour, 100 mL of water was added to the reaction solution, extracted with ethyl acetate (100 mL×3), and the combined organic phases were then added with saturated common salt Washed with water (100 mL), finally the organic phase was dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0～4/1) Compound 18-2 was obtained. 1H NMR (400 MHz, CDCl ₃ ) δ 1.46-1.49 (m, 9H), 1.61-1.69 (m, 4H), 1.77-1.87 (m, 2H), 1.97-2.04 (m, 2H) ), 4.27-4.32(m, 2H), 4.44-4.69(m, 1H), 5.78-5.94(s, 1H), 6.53-6.69(s, 1H).

Step 2: Compound 18-2 (0.6 g, 1.65 mmol) was dissolved in dioxane (10 mL), 1-4 (412.46 mg, 1.81 mmol), cesium carbonate (1.07 g, 3.30 mmol) and [( 2-Di-cyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino- 1,1'-biphenyl)] palladium(II) mesylate (74.74 mg, 82.45 μmol) was replaced with nitrogen three times and then heated to 100° C. and stirred under nitrogen protection for 16 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-5/1) to obtain compound 18-3. MS ESI calculated: C ₂₇ H ₄₂ N ₈ O ₃ Si[M+H] ⁺ 555, found 555. ¹ H NMR (400MHz, CD ₃ OD) δ 0.00-0.01 (m, 9H), 0.91-0.97 (m, 2H), 1.25-1.29 (m, 2H), 1.48-1.49 (m, 9H), 1.60- 1.73(m,4H),1.77-1.83(m,2H),1.94-2.03(m,2H),2.26-2.29(s,3H),4.25-4.37(m,2H),4.88-5.03(m,1H ), 5.42(s, 2H), 6.17-6.25(s, 1H), 6.34-6.42(s, 1H), 7.40-7.51(s, 1H).

Step 3: Compound 18-3 (0.35 g, 630.91 μmol) was dissolved in DMF (10 mL) at 25° C., water (0.5 mL), triethylamine (363.50 mg, 3.59 mmol, 0.5 mL) and Selenium powder (150 mg, 1.85 mmol) was replaced with carbon monoxide three times and reacted at 90 ° C under 15 psi pressure for 1.5 hours, 50 mL of ammonium chloride aqueous solution was added to the reaction solution, and extracted with dichloromethane (50 mL × 3), the combined organic The phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 18-4.

Step 4: Compound 18-4 (0.4 g, 629.19 μmol) was dissolved in ethanol (4 mL) at 25° C., 6-5 (58.22 mg, 629.19 μmol) was added, and the mixture was stirred at 80° C. for 1 hour. The reaction solution was directly concentrated under reduced pressure to obtain crude product 18-5. MS ESI _calcd for _{C30H46N8O3SeSi [ M} +H] ⁺ 675, found 675.

Step 5: Compound 18-5 (350 mg, 519.46 μmol) was dissolved in ethyl acetate solution of hydrochloric acid (4 M, 5 mL) and stirred at 25° C. for 2 hours. The reaction solution was directly filtered to obtain the crude hydrochloride of compound 18-6 as the filter cake. MS ESI _calcd for _C19H24N8Se [ _M +H] ⁺ 445, found 445.

Step 6: The crude hydrochloride of 18-6 (100 mg, 193.68 μmol) was dissolved in methanol (10 mL), N,N-diisopropylethylamine (75.1 mg, 581.0 μmol, 101.20 μL) was added, and the mixture was After stirring at 25°C for 10 minutes, acrylonitrile (1-10) (20.55 mg, 387.35 μmol, 25.69 μL) was added, and the mixture was stirred at 25°C for 16 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 14%-44% , 10 minutes) to obtain the trifluoroacetate salt of compound 18-7. MS ESI calculated for C ₂₂ H ₂₇ N ₉ Se[M+H] ⁺ 498, found 498, ¹ H NMR (400 MHz, CD ₃ OD) δ 2.04-2.13 (m, 2H), 2.21-2.28 (m, 2H), 2.31-2.47(m, 7H), 2.57-2.61(s, 3H), 3.07-3.12(m, 2H), 3.45-3.53(m, 2H), 4.18-4.28(m, 2H), 4.56- 4.73(m, 1H), 5.87-6.04(s, 1H), 6.66-6.88(s, 1H), 8.07-8.13(s, 1H).

Example 19

Step 1: The crude hydrochloride of 18-6 (30 mg, 44.68 μmol) was dissolved in DMF (0.5 mL), triethylamine (22.61 mg, 223.40 μmol, 31.09 μL) was added, the mixture was stirred for 0.5 hours, and then added to the reaction DMF (0.5 mL) was added to the liquid and cooled to 0 °C, stirred over 0.5 h, 3-1 (9.68 mg, 53.62 μmol) was added slowly, the mixture was warmed to 20 °C and stirred for 3 h. LC-MS showed that the starting material was consumed and the main peak was the product peak. Saturated aqueous ammonium chloride solution (20 mL) was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, Filtration and concentration under reduced pressure gave the crude product, which was preparatively separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 28%- 58%, 10 min) to give the trifluoroacetate salt of compound 19-1. MS ESI calculated for C ₂₃ H ₂₈ N ₁₀ O ₂ SSe[M+H] ⁺ 589, found 589, ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.24-1.36 (m, 1H), 1.82 (m, 2H ), 1.87-1.94(m, 2H), 2.10-2.17(m, 2H), 2.22-2.27(m, 2H), 2.40(s, 3H), 2.57(s, 3H), 3.39-3.48(m, 1H ),3.87-4.00(m,1H),4.10-4.18(m,4H),4.32(m,2H),5.86(s,1H),6.80(s,1H),7.74(d,1H),7.96( m, 1H), 9.01-9.31 (m, 1H).

Example 20

Step 1: At 20°C, diisopropylethylamine (2.50 g, 19.32 mmol) and compound 20-1 (2 g, 9.66 mmol) were added to tetrahydrofuran (40 mL), 1-2 (2.19 g, 9.66 mmol) It was dissolved in tetrahydrofuran (10 mL), dropped into the former system at 0°C, and reacted at 20°C for 18 hours. The reaction solution was diluted with 150 mL of water, extracted with 120 mL of ethyl acetate (40 mL×3), the combined organic layers were washed with 40 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 1/1) to give compound 20-2.

Step 2: Compound 20-2 (2.15 g, 5.42 mmol) was dissolved in methanol (20 mL) and water (6 mL), lithium hydroxide (454.63 mg, 10.83 mmol) was added, and the reaction was carried out at 20°C for 2 hours. The pH of the reaction solution was adjusted to 4 with 0.5M aqueous hydrochloric acid solution, the reaction solution was diluted with 100 mL of water, extracted with 120 mL of ethyl acetate (40 mL×3), the combined organic layers were washed with brine 40 mL, dried over sodium sulfate, filtered and reduced in pressure Concentration gave compound 20-3. MS ESI calculated: _C17H23ClN4O4 [M ₊ H] ₊₃₈₃ , found ₃₈₃ .

Step 3: Compound 20-3 (1.9 g, 4.96 mmol) was dissolved in dichloromethane (40 mL), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.14 g, 5.96 mmol) and 1-hydroxybenzotriazole (804.70 mg, 5.96 mmol), reacted at 20° C. for 0.5 hour. Then, methoxymethylamine hydrochloride (726.12 mg, 7.44 mmol) and triethylamine (2.01 g, 19.85 mmol) were added, and the reaction solution was reacted at 20° C. for 16 hours. The reaction solution was diluted with 100 mL of water, extracted with 100 mL of ethyl acetate (50 mL×2), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 3/1) to give compound 20-4. MS ESI calculated: _C19H28ClN5O4 [M ₊ H] ₊₄₂₆ , found ₄₂₆ .

Step 4: Compound 20-4 (1.80 g, 4.24 mmol) was dissolved in tetrahydrofuran (30 mL), methylmagnesium bromide ether solution (3M, 3.11 mL,) was added dropwise at 0°C, and the reaction was carried out at 0°C for 1 hour. 20 mL of saturated ammonium chloride solution was added to quench the reaction, the reaction solution was diluted with 60 mL of water, extracted with 60 mL of ethyl acetate (30 mL × 2), the combined organic layers were washed with 30 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. Silica gel column purification (petroleum ether/ethyl acetate=10/1 to 4/1) gave compound 20-5. ¹ H NMR (400MHz, CDCl ₃ ) δ 1.47(s, 9H), 1.52-1.93(m, 4H), 2.05(s, 4H), 2.61(s, 3H), 4.14-4.37(m, 2H), 4.37-4.73(m,1H),5.37-5.74(m,1H),6.82(s,1H)

Step 5: Compound 20-5 (660 mg, 1.73 mmol) was dissolved in dichloromethane (20 mL), triethylamine (1.05 g, 10.40 mmol) was added, and tert-butyldimethylsilyltrifluoromethyl was added dropwise Sulfonate (1.37 g, 5.20 mmol) was reacted at 15°C for 1 hour. The reaction solution was diluted with dichloromethane 60 mL, and was diluted with saturated aqueous sodium bicarbonate solution 60 mL (30 mL×2), water (20 mL) and brine 30 mL, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 20-6. MS ESI calculated: _{C30H53ClN4O3Si} [ _{M +} H] ₊₆₀₉ , found 609.

Step 6: Compound 20-6 (1.14 g, 1.87 mmol) was dissolved in tetrahydrofuran (30 mL) and water (5 mL), bromosuccinimide (332.95 mg, 1.87 mmol) was added, and the reaction was carried out at 15°C for 2 hours. The reaction solution was diluted with 100 mL of water, extracted with 100 mL of ethyl acetate (50 mL×2), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 20-7. MS ESI calculated: _{C18H24BrClN4O3 [ M +} H] ⁺ 459, found 459.

Step 7: Compound 20-7 (650 mg, 1.13 mmol) was dissolved in methanol (25 mL), compound 12-2 (138.18 mg, 1.13 mmol) was added at 0-5 °C, and the reaction was carried out at 15 °C for 0.5 hour. Then BOC acid anhydride (296.56 mg, 1.36 mmol), 2,6-lutidine (13.83 mg, 113.24 μmol) and triethylamine (458.33 mg, 4.53 mmol) were added, and the reaction was carried out at 15° C. for 16 hours. The reaction solution was diluted with 100 mL of water, extracted with 90 mL of ethyl acetate (30 mL×3), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 5/1 to 3/1) to give compound 20-8. MS _ESI calculated: _{C20H26ClN5O2Se} [ _{M +} H] ⁺ 484, found 484.

Step 8: Compound 20-8 (100 mg, 205.03 μmol), 8-9 (48.53 mg, 246.03 μmol) were added to dioxane (3 mL), followed by [(2-di-cyclohexylphosphino-3, 6-Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methane Palladium(II) sulfonate methanesulfonate (18.59 mg, 20.50 μmol), cesium carbonate (133.60 mg, 410.05 μmol), reacted at 80° C. under nitrogen atmosphere for 3 hours. The reaction solution was diluted with 50 mL of water, and extracted with 40 mL (20 mL×2) of ethyl acetate. The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 5/1 to 1/1) purification to give compound 20-9. MS ESI calculated: _C29H40N8O4Se [M ₊ H] ₊₆₄₅ , found ₆₄₅ .

Step 9: Compound 20-9 (65 mg, 100.99 μmol) was dissolved in methanol (1 mL), ethyl acetate hydrochloride (4 M, 2 mL) was added, and the reaction was carried out at 50° C. for 2 hours. The reaction solution was concentrated under reduced pressure to obtain compound 20-10. MS _ESI calculated: _C19H24N8Se [ _M +H] ⁺ 444, found 444.

Step 10: Compound 20-10 (50 mg, 104.20 μmol) was dissolved in methanol (2 mL), diisopropylethylamine (67.33 mg, 520.98 μmol) and acrylonitrile (0.22 g, 4.15 mmol) were added at 15° C. React for 2 hours. The reaction solution was added with 5 mL of water and 5 mL of methanol, stirred for 15 minutes, and filtered to obtain a filter cake. The filter cake was slurried by adding 5 mL of methanol, and filtered to obtain compound 20-11. MS _ESI calculated: _C22H27N9Se [M+H] ₊₄₉₈ , found 498. ¹ H NMR (400MHz, DMSO-d ₆ )δ1.52(d,J=10.4Hz,2H),1.71(d,J=6.8Hz,2H),1.80(s,2H),1.92(d,J= 2.8Hz, 2H), 2.20(s, 3H), 2.60(m, 4H), 2.73(s, 3H), 3.25-3.32(m, 2H), 4.13-4.43(m, 1H), 6.60(s, 2H) ),6.94-7.24(m,1H),8.36-9.11(m,2H),11.72(s,1H).

Example 21

Step 1: Compound 20-7 (600 mg, 1.05 mmol) was dissolved in methanol (25 mL), compound 8-6 (201.84 mg, 1.05 mmol) was added at 0-5 °C, and the reaction was carried out at 15 °C for 0.5 hour. Then (Boc) ₂ O (273.75 mg, 1.25 mmol), 2,6-lutidine (12.77 mg, 104.53 μmol) and triethylamine (423.07 mg, 4.18 mmol) were added, and the reaction was carried out at 15° C. for 16 hours. The reaction solution was diluted with 100 mL of water, extracted with 90 mL of ethyl acetate (30 mL×3), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 5/1 to 3/1) to obtain compound 21-1. MS ESI calculated: _{C23H31ClN6O3Se [ M} +H] ⁺ ₅₅₅ , found 555.

Step 2: Compound 21-1 (250 mg, 451.31 μmol), 8-9 (106.82 mg, 541.57 μmol) were added to dioxane (8 mL), followed by [(2-di-cyclohexylphosphino-3, 6-Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methane Palladium(II) sulfonate mesylate (40.91 mg, 45.13 μmol) and cesium carbonate (294.09 mg, 902.62 μmol) were reacted at 80° C. under nitrogen atmosphere for 3 hours. The reaction solution was diluted with 50 mL of water, and extracted with 40 mL (20 mL×2) of ethyl acetate. The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 5/1 to 1/1) to obtain compound 21-2. MS _ESI calculated: _C32H45N9O5Se [M+ _H ] ⁺ ₇₁₆ , found 716.

Step 3: Compound 21-2 (220 mg, 270.88 μmol,) was dissolved in methanol (2 mL), ethyl acetate hydrochloride (4 M, 6 mL) was added, and the reaction was carried out at 40° C. for 2 hours. The reaction solution was concentrated under reduced pressure to obtain compound 21-3. MS _ESI calculated: _C22H29N9Se [M+H] ⁺ ₅₁₆ , found 516.

Step 4: Compound 21-3 (150 mg, 272.26 μmol) was dissolved in methanol (5 mL), diisopropylethylamine (175.93 mg, 1.36 mmol) and acrylonitrile (28.89 mg, 544.52 μmol) were added at 15° C. React for 2 hours. The reaction solution was added with 10 mL of water and 10 mL of methanol, stirred for 15 minutes, and filtered to obtain a filter cake, which was slurried by adding 10 mL of methanol, and filtered to obtain compound 21-4. MS ESI calculated: C ₂₅ H ₃₂ N ₁₀ OSe[M+H] ⁺ 569, found 569. ¹ H NMR (400MHz, DMSO-d ₆ )δ1.40-1.56(m, 2H), 1.70(d, J=7.2Hz, 2H), 1.77(s, 2H), 1.86-1.97(m, 2H), 2.20(s, 3H), 2.59(m, 4H), 3.29(s, 2H), 3.43(s, 4H), 3.72(d, J＝4.40Hz, 4H), 4.10-4.37(m, 1H), 6.49 (d, J=14.4Hz, 2H), 7.03(d, J=2.4Hz, 1H), 7.97(s, 1H), 8.55(s, 1H), 11.70(s, 1H)

Example 22

Step 1: 22-1 (1.2 g, 2.48 mmol) was dissolved in ethanol (50 mL), 22-2 (580.83 mg, 2.98 mmol) was added, and the mixture was stirred at 80° C. for 1 hour. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was diluted with 50 mL of tetrahydrofuran, triethylamine (502.31 mg, 4.96 mmol) and di-tert-butyl dicarbonate (541.70 mg, 2.48 mmol) were added, and the mixture was stirred at 80° C. for 1 hour. The reaction solution was diluted with 100 mL of water, extracted with ethyl acetate (50 mL×2), washed with saturated brine (50 mL×2), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. Column chromatography (SiO ₂ , petroleum ether: ethyl acetate=20/1～3/1) was used for purification to obtain compound 22-3. MS _ESI calcd for _{C23H29ClN4O4Se} [M ₊ H] ₊ 540, found 540 ^.

Step 2: Dissolve 22-3 (280 mg, 518.60 μmol) in tetrahydrofuran (6 mL), add a solution of lithium hydroxide monohydrate (65.29 mg, 1.56 mmol) in water (2 mL), and stir at 35° C. for 16 hours. The reaction solution was adjusted to pH=4-5 with 1M dilute hydrochloric acid, and concentrated to obtain the crude product. The crude product was soaked in tetrahydrofuran (10 mL) and methanol (5 mL), filtered, and the filtrate was concentrated to obtain compound 22-4. MS _ESI calcd for _{C21H25ClN4O4Se} [M ₊ H] ₊ 512, found 512 ^.

Step 3: 22-4 (160 mg, 312.59 μmol) was dissolved in acetonitrile (4 mL), 22-5 (35.69 mg, 625.17 μmol) and N,N-diisopropylethylamine (161.59 mg, 1.25 mmol) were added 2-(7-azobenzotriazole)-N,N,N,N-tetramethylurea hexafluorophosphate (130.74mg, 343.84μmol) was added at 30°C under stirring for 16 hours. The reaction solution was diluted with water (10 mL), extracted with ethyl acetate (10 mL×2), washed with saturated brine (20 mL×2), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was passed through the column layer Compound 22-6 was obtained by purification (SiO ₂ , petroleum ether: ethyl acetate=5/1～1/1). MS ESI _calcd for _{C24H30ClN5O3Se [ M} +H] ⁺ 551, found 551.

Step 4: Compound 22-6 (150 mg, 272.26 μmol), 8-9 (64.44 mg, 326.71 μmol), cesium carbonate (177.42 mg, 544.52 μmol) and [(2-di-cyclohexylphosphino-3,6 -Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methanesulfonic acid Palladium (II) methanesulfonate (24.68 mg, 27.23 μmol) was dissolved in dioxane (5 mL), replaced with nitrogen three times and then heated to 100° C., and stirred for 16 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=4/1-1/2) to obtain compound 22-7. MS ESI calculated: C ₃₃ H ₄₄ N ₈ O ₅ Se[M+H] ⁺ 712, found 712.

Step 5: Compound 22-7 (100 mg, 140.51 μmol) was dissolved in ethyl acetate (2 mL), ethyl acetate solution of hydrochloric acid (4 M, 2 mL) was added, and the mixture was stirred at 30° C. for 16 hours. The reaction solution was directly filtered to obtain the crude hydrochloride of compound 22-8 as the filter cake. MS _ESI calculated for _C23H28N8OSe [M+H] ⁺ ₅₁₂ , found 512.

Step 6: The crude hydrochloride of 22-8 (70 mg, 136.86 μmol, HCl) was dissolved in methanol (5 mL), N,N-diisopropylethylamine (88.44 mg, 684.29 μmol), and acrylonitrile were added. (1-10) (21.79 mg, 410.57 μmol), stirred at 25° C. for 16 hours. LC-MS showed that the starting material was completely consumed and the main peak was the product peak. 10 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, the crude product After preparative separation by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; B(ACN)%: 12%-42%, 10 minutes), Ammonia water was adjusted to pH=8, extracted with ethyl acetate (10 mL×3), the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 22-9. MS ESI calculated for C ₂₆ H ₃₁ N ₉ OSe[M+H] ⁺ 565, found 565, ¹ H NMR (400 MHz, DMSO-d6) δ 0.61-0.67 (m, 2H), 0.69-0.75 (m, 2H), 1.50(m, 2H), 1.73(m, 2H), 1.76-1.85(m, 2H), 1.87-1.97(m, 2H), 2.19(s, 3H), 2.52(m, 1H), 2.56 -2.65(m, 4H), 2.86(m, 1H), 3.31(s, 3H), 4.08-4.29(m, 1H), 6.30(s, 1H), 6.33-6.49(m, 1H), 6.50-6.67 (m, 1H), 8.20 (s, 1H), 8.85 (s, 1H), 11.56-11.88 (m, 1H).

Example 23

Step 1: Dissolve 22-3 (80 mg, 156.29 μmol) in acetonitrile (4 mL), add 23-1 (32.50 mg, 312.59 μmol) and N,N-diisopropylethylamine (80.80 mg, 625.17 μmol) 2-(7-azobenzotriazole)-N,N,N,N-tetramethylurea hexafluorophosphate (65.37mg, 171.92μmol) was added at 50°C under stirring for 16 hours. The reaction solution was diluted with water (10 mL), extracted with ethyl acetate (10 mL×2), washed with saturated brine (20 mL×2), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was passed through the column layer Precipitation (SiO ₂ , petroleum ether: ethyl acetate=3/1～1/1) and purification to obtain compound 23-2. MS _{ESI calcd} for _{C22H28ClN5O3Se} [ _M +H] ⁺ 525, found 525.

Step 2: Compound 23-2 (60 mg, 114.31 μmol), 8-9 (27.05 mg, 137.17 μmol), cesium carbonate (74.49 mg, 228.61 μmol) and [(2-di-cyclohexylphosphino-3,6 -Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methanesulfonic acid Palladium(II) methanesulfonate (20.72 mg, 22.86 μmol) was dissolved in dioxane (5 mL), replaced with nitrogen three times, then heated to 100° C., and stirred under nitrogen protection for 16 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=4/1-1/1) to obtain compound 23-3. MS ESI calculated value: C ₃₁ H ₄₂ N ₈ O ₅ Se[M+H] ⁺ 686, found 686.

Step 3: Compound 23-3 (40 mg, 58.34 μmol) was dissolved in ethyl acetate (2 mL), ethyl acetate solution of hydrochloric acid (4 M, 2 mL) was added, and the mixture was stirred at 30° C. for 3 hours. The reaction solution was directly filtered to obtain the crude hydrochloride of compound 23-4 as the filter cake. MS _ESI calculated for _C21H26N8OSe [M+H] ⁺ ₄₈₅ , found 485.

Step 4: The crude hydrochloride of 23-4 (60 mg, 123.60 μmol) was dissolved in methanol (5 mL), N,N-diisopropylethylamine (79.87 mg, 617.99 μmol), and acrylonitrile (1 -10) (19.68 mg, 370.80 μmol), stirred at 25° C. for 1 hour. 10 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure The crude product was obtained, and the crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; B(ACN)%: 10%-40%, 10 minutes), adjusted to pH=8 with aqueous ammonia, extracted with ethyl acetate (10 mL×3), the combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 23-5. MS ESI calculated for C ₂₄ H ₂₉ N ₉ OSe[M+H] ⁺ 539, found 539, ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 1.51 (m, 2H), 1.73 (m, 2H), 1.79(s, 2H), 1.91(m, 2H), 2.19(s, 3H), 2.55-2.65(m, 5H), 2.81(d, J=4.75Hz, 3H), 3.31(s, 3H), 4.07 -4.31(m,1H),6.29(s,1H),6.37(s,1H),6.60(s,1H),8.23(s,1H),8.84(s,1H),11.58-11.86(s,1H) )

Example 24

Step 1: Dissolve 22-3 (200 mg, 370.43 μmol) in tetrahydrofuran (5 mL), add a solution of diisobutylaluminum hydride in toluene (1 M, 1.18 mL) at -65 °C, and stir at 0 °C for 2 hours . At 0°C, saturated potassium sodium tartrate (5 mL) was added to the reaction solution to quench for 0.5 hours, and then saturated brine (10 mL) was added to dilute, extracted with ethyl acetate (20 mL×2), and the organic phase was washed with saturated brine ( 20 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 24-1. MS _ESI calcd for _{C21H27ClN4O3Se} [ _{M +} H] ⁺ 498, found 498.

Step 2: Dissolve 24-1 (180 mg, 361.54 μmol) in dichloromethane (5 mL), add Dess Martin Periodane (230.01 mg, 542.30 μmol l), and stir at 20° C. for 1 hour. Saturated sodium sulfite (5 mL) and saturated sodium bicarbonate (5 mL) were added to the reaction solution, stirred at 20° C. for 10 minutes to quench, then added water (10 mL) to dilute, extracted with dichloromethane (20 mL×2), and the organic phase was saturated with Washed with brine (20 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=10/1～3/1) to obtain the compound 24-2. MS _ESI calcd for _{C21H25ClN4O3Se} [ _{M +} H] ⁺ 496, found 496.

Step 3: Dissolve 24-2 (170 mg, 310.20 μmol) in tetrahydrofuran (5 mL), add elemental iodine (393.66 mg, 1.55 mmol) and ammonia water (434.85 mg, 3.10 mmol, 25%), stir at 30° C. for 16 Hour. Saturated sodium sulfite (5 mL) and saturated sodium bicarbonate (5 mL) were added to the reaction solution, stirred at 20° C. for 10 minutes to quench, then added water (10 mL) to dilute, extracted with dichloromethane (20 mL×2), and the organic phase was saturated with Washed with brine (20 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=10/1～3/1) to obtain the compound 24-3. MS ESI _calcd for _{C21H24ClN5O2Se} [ _{M +} H] ⁺ 496, found 493.

Step 4: Compound 24-3 (100 mg, 198.84 μmol), 8-9 (43.14 mg, 218.72 μmol), cesium carbonate (194.36 mg, 596.52 μmol) and [(2-di-cyclohexylphosphino-3,6 -Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methanesulfonic acid Palladium (II) methanesulfonate (18.02 mg, 19.88 μmol) was dissolved in dioxane (4 mL), replaced with nitrogen three times, then heated to 90° C., and stirred for 2 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate = 10/1 to 3/1) to obtain compound 24-4. MS ESI calculated: C ₃₀ H ₃₈ N ₈ O ₄ Se[M+H] ⁺ 654, found 654.

Step 5: Compound 24-4 (100 mg, 151.00 μmol) was dissolved in ethyl acetate (2 mL), ethyl acetate solution of hydrochloric acid (4 M, 2 mL) was added, and the mixture was stirred at 45° C. for 2 hours. The reaction solution was directly concentrated to obtain the crude hydrochloride of compound 24-5. MS _ESI calculated for _C20H22N8Se [ _M +H] ⁺ 453, found 453.

Step 6: The crude hydrochloride of 24-5 (70 mg, 154.39 μmol) was dissolved in methanol (2 mL), N,N-diisopropylethylamine (99.77 mg, 771.94 μmol) was added, and acrylonitrile (1 -10) (24.58 mg, 463.17 μmol), stirred at 20° C. for 1 hour. LC-MS showed that the starting material was consumed and the main peak was the product peak. 10 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (20 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 13%-43% , 10 min) to obtain the trifluoroacetate salt of compound 24-6. MS ESI calculated for C ₂₃ H ₂₅ N ₉ Se[M+H] ⁺ 506, found 506, ¹ H NMR (400 MHz, CD ₃ OD) δ 2.05 (m, 3H), 2.33 (m, 2H), 2.36 (s,3H),2.38(m,1H),2.42(m,2H),3.07-3.11(m,2H),3.50(m,2H),4.22(s,2H),4.32(m,1H), 6.00(s, 1H), 6.67(s, 1H), 6.72(s, 1H), 9.27(s, 1H).

Example 25

Step 1: 25-1 (10g, 109.73mmol) was added to absolute ethanol (30mL), then methyl iodide (18.38g, 129.48mmol) was added, stirred at 80°C for 4 hours in a stuffy tank, the solid was precipitated and directly filtered to obtain the compound The hydriodate of 25-2. ¹ H NMR (400 MHz, D ₂ O) δ 2.60 (s, 3H).

Step 2: at 25 ° C, add selenium powder (6.95g, 85.81mmol) to ethanol (140mL), then add sodium borohydride (3.57g, 94.39mmol) in batches, after the black selenium powder disappears completely, add Sodium carbonate (4.55 g, 42.90 mmol) and the hydroiodide salt of 25-2 (10 g, 42.90 mmol), and warmed to 25 °C and stirred for 16 hours. The reaction solution was quenched by adding 20 mL of glacial acetic acid, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to column chromatography (SiO ₂ , petroleum ether: ethyl acetate=2/1～0/1 and then dichloromethane: methanol=20/1). 1～10/1) purification to obtain compound 25-3. ¹ H NMR (400 MHz, DMSO-d6) δ 3.80 (s, 2H), 8.26 (s, 1H), 8.57 (s, 1H), 9.67 (s, 1H).

Step 3: 25-3 (4 g, 28.98 mmol) and 1-1 (4.01 g, 23.18 mmol) were added to trifluoroacetic acid (50 mL), and the temperature was raised to 80°C and stirred for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the crude product. The crude product was diluted with ethyl acetate (100 mL), the pH was adjusted to 7-8 with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate (50 mL×3), and the organic phase was diluted with saturated brine (50 mL). ×2) Washed, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=4/1～3/2) to obtain compound 25-4. MS ESI _calcd for _C7H4Cl2N4Se [M ₊ H] ₊ 294, found 294 ^.

Step 4: 25-4 (1 g, 2.99 mmol) was added to DMF (20 mL) followed by triethylamine (605.76 mg, 5.99 mmol), 4-dimethylaminopyridine (73.13 mg, 598.64 μmol) and dicarbonic acid Di-tert-butyl ester (979.88 mg, 4.49 mmol) was stirred at 25°C for 1 hour. Water (50 mL) was added to dilute, extracted with ethyl acetate (30 mL×3), the organic phase was washed with saturated brine (50 mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was subjected to column chromatography ( SiO ₂ , petroleum ether: ethyl acetate=20/1～5/1) and purified to obtain compound 25-5. MS ESI _calcd for _{C12H12Cl2N4O2Se} [M ₊ H] _{+ 394} , found 394 ^.

Step 5: Compound 25-5 (1.2 g, 3.04 mmol), 1-2 (826.90 mg, 3.65 mmol), tris(dibenzylideneacetone)dipalladium (278.82 mg, 304.48 μmol), 4,5-bis (Diphenylphosphorus)-9,9-dimethylxanthene (352.35 mg, 608.96 μmol) and cesium carbonate (1.98 g, 6.09 mmol) were added to dioxane (40 mL), and the temperature was raised to Stir at 100°C for 16 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was subjected to column chromatography (SiO ₂ , petroleum ether: ethyl acetate=10/1～3/1) and then to high performance liquid chromatography for preparative separation (column: Waters Xbridge C18 150* 50mm*10μm; mobile phase: [water (10mM ammonium bicarbonate)-ACN]; B(ACN)%: 56%-86%) was purified to obtain compound 25-6. MS ESI calculated: _{C24H33ClN6O4Se [ M +} H] ⁺ 584, found 584.

Step 6: Compound 25-6 (120 mg, 182.89 μmol), 8-9 (43.29 mg, 219.46 μmol), cesium carbonate (178.76 mg, 548.66 μmol) and [(2-di-cyclohexylphosphino-3,6 -Dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)]methanesulfonic acid Palladium (II) methanesulfonate (16.58 mg, 18.29 μmol) was added to dioxane (5 mL), replaced with nitrogen three times and then heated to 90° C. and stirred for 3 hours under nitrogen protection. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=10/1-2/1) to obtain compound 25-7. MS ESI calculated: C ₃₃ H ₄₇ N ₉ O ₆ Se[M+H] ⁺ 745, found 745.

Step 7: Compound 25-7 (80 mg, 107.42 μmol) was dissolved in ethyl acetate (2 mL), ethyl acetate solution of hydrochloric acid (4 M, 2 mL) was added, and the mixture was stirred at 45° C. for 2 hours. The reaction solution was directly concentrated to obtain the crude hydrochloride of compound 25-8. MS _{ESI calcd} for _C18H23N9Se [M+H] ⁺ 444, found 444.

Step 8: The crude hydrochloride of 25-8 (50 mg, 103.98 μmol) was dissolved in methanol (2 mL), N,N-diisopropylethylamine (67.19 mg, 519.91 μmol) was added, and acrylonitrile (1 -10) (16.55 mg, 311.94 μmol), stirred at 20° C. for 1 hour. 10 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (20 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 9%-39% , 10 min) to obtain the trifluoroacetate salt of compound 25-9. MS ESI calculated for C ₂₁ H ₂₆ N ₁₀ Se[M+H] ⁺ 497, found 497, ¹ H NMR (400 MHz, CD ₃ OD) δ 2.10 (m, 3H), 2.33 (m 1H), 2.36 ( s,3H),2.39-2.40(m,1H),2.43(m,3H),3.10(m,3H),3.51(m,3H),4.24(m,3H),5.92(s,1H),6.53 (s, 1H), 6.56 (s, 1H).

Example 26

Step 1: At 20°C, the hydrochloride salt of compound 18-6 (60 mg, 89.36 μmol) was dissolved in DMF (0.5 mL), potassium carbonate (24.70 mg, 178.72 μmol) and 4-1 (24.02 mg, 107.23 μmol, 12.57 μL), and the mixture was stirred at 50° C. for 16 hours. The reaction solution was directly concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 16%-46%, 10 minutes) to obtain compound 26 -1 of the trifluoroacetate salt. MS ESI calculated: C ₂₂ H ₂₇ F ₃ N ₈ Se[M+H] ⁺ 541, found 541, ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 1.83 (m, 2H), 2.07-2.11 ( m, 2H), 2.23(s, 3H), 2.30-2.34(m, 2H), 2.44(s, 3H), 2.81-2.99(m, 2H), 3.17-3.30(m, 2H), 4.14(m, 3H), 4.25-4.49(m, 2H), 6.33(m, 1H), 6.70(s, 1H), 7.62-7.84(m, 1H), 7.96(s, 1H), 9.46-9.65(m, 1H) ,9.69(m,1H).

Example 27

Step 1: Compound 27-1 (1.8 g, 12.28 mmol) was dissolved in tetrahydrofuran (10 mL) and saturated sodium bicarbonate solution (10 mL) at 0 °C, CbzCl (2.30 g, 13.50 mmol, 1.92 mL) was added, and Stir at 20°C for 16 hours. 10 mL of water was added to the reaction solution, and extracted with dichloromethane (20 mL × 3). The combined organic phases were washed with 0.5 M aqueous hydrochloric acid (5 mL × 3) and saturated brine (5 mL × 3) in turn. Finally, the organic phase was washed with Dry over anhydrous sodium sulfate, filter and concentrate under reduced pressure to obtain crude product, which is purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=8/1～3/1) to obtain compound 27-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.77-1.96 (m, 4H), 2.84 (m, 1H), 3.46 (m, 2H), 3.73 (m, 2H), 5.14 (s, 2H), 7.30- 7.44 (m, 5H).

Step 2: Compound 27-2 (950 mg, 3.89 mmol), selenium (629.80 mg, 7.78 mmol, 617.45 μL) were dissolved in N,N-DMF (10 mL) and water (1 mL), the mixture was vented under vacuum , and the gas was replaced three times with carbon monoxide, and the reaction mixture was stirred at 80° C. for 16 hours under carbon monoxide gas (50 psi). The reaction solution was directly concentrated under reduced pressure to remove the solvent, 20 mL of water was added, and extracted with ethyl acetate (20 mL×3), the combined organic phases were washed with saturated brine (20 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, Filtration and concentration under reduced pressure gave crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=4/1˜1/2) to obtain compound 27-3. ¹ H NMR (400MHz, CDCl ₃ )δ1.67-1.83(m,3H), 1.94(m,2H), 2.79-2.87(m,2H), 4.21-4.41(m,2H), 5.13(m,2H) ), 7.32-7.40 (m, 5H), 7.77 (m, 1H), 8.51 (m, 1H).

Step 3: Compound 27-3 (1.35 g, 4.15 mmol) was dissolved in methanol (15 mL), 8-4 (858.58 mg, 3.19 mmol) was added, and the mixture was stirred at 20° C. for 16 hours. The reaction solution was directly concentrated under reduced pressure to obtain the crude product, and the crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=4/1～3/1) to obtain compound 27-4. MS ESI _calcd for _{C21H19Cl2N3O2Se} [ _{M +} H] ₊ 496, found 496 ^.

Step 4: Compound 27-4 (500 mg, 1.01 mmol), 1-2 (228.58 mg, 1.01 mmol), (±)-2,2-bis(diphenylphosphino)-11-binaphthyl (62.89 mg, 101.00 μmol), palladium acetate (22.68 mg, 101.00 μmol) and cesium carbonate (658.15 mg, 2.02 mmol) were dissolved in dioxane (12 mL), the mixture was purged with nitrogen three times, and stirred at 90° C. for 16 hours. 30 mL of water was added to the reaction solution, extracted with ethyl acetate (30 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure The crude product was obtained, and the crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=8/1～1/1) to obtain compound 27-5. MS _{ESI calcd} for _{C33H40ClN5O4Se} [M ₊ H] ⁺ 686, found 686.

Step 5: Compound 27-5 (500 mg, 729.81 μmol), 8-9 (158.34 mg, 802.79 μmol), methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2', 4',6'-Triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (BrettPhos Pd G3) (66.16mg, 72.98 μmol) and cesium carbonate (475.57 mg, 1.46 mmol) were dissolved in dioxane (12 mL), the mixture was purged with nitrogen three times, and stirred at 90° C. for 16 hours. 40 mL of water was added to the reaction solution, extracted with dichloromethane (30 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure The crude product was obtained, and the crude product was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=5/1～1/1) to obtain compound 27-6. MS ESI _calcd for _{C42H54N8O6Se [} M+H] ⁺ ₈₄₇ , found 847.

Step 6: Compound 27-6 (380 mg, 449.23 μmol) was dissolved in methanol (1 mL), ethyl acetate solution of hydrochloric acid (4 M, 4 mL) was added, and the mixture was stirred at 20° C. for 16 hours. The reaction solution was directly spin-dried to obtain the crude hydrochloride of compound 27-7. MS ESI _calcd for _C32H38N8O2Se [M ₊ H] ⁺ ₆₄₇ , found 647.

Step 7: The crude hydrochloride of 27-7 (300 mg, 417.49 μmol) was dissolved in methanol (3 mL), N,N-diisopropylethylamine (161.87 mg, 1.25 mmol, 218.16 μL) was added, followed by Compound acrylonitrile (1-10) (33.23 mg, 626.24 μmol, 41.54 μL) was stirred at 20° C. for 2 hours. LC-MS showed that the starting material was consumed and the main peak was the product peak. The reaction solution was directly concentrated under reduced pressure to obtain a crude product, which was prepared and separated by high performance liquid chromatography (column: Phenomenex luna C18 150*40mm*15 μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 18% -48%, 11 min) to give compound 27-8 as the trifluoroacetate salt. MS ESI _calcd for _C35H41N9O2Se [ _{M +} H] ⁺ 700, found 700.

Step 8: The trifluoroacetate salt of 27-8 (90 mg, 128.81 μmol) was dissolved in hydrobromic acid (0.3 mL) and stirred at 15° C. for 16 hours. The reaction solution was filtered to obtain a crude product, which was prepared and separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 8%-38% , 10 min) to obtain the trifluoroacetate salt of compound 27-9. MS _{ESI calcd} for _C27H35N9Se [M+H] ⁺ 566, found 566. ¹ H NMR (400MHz, DMSO-d ₆ )δ1.84-1.95(m,4H), 2.14(m,2H), 2.19-2.24(m,2H), 2.25(m,3H), 2.28(m,2H) ),3.06-3.13(m,4H),3.40(m,6H),4.03-4.25(m,4H),6.02(s,1H),6.57(s,1H),6.78(s,1H),8.46- 8.64 (m, 1H), 8.74 (m, 1H), 8.84 (m, 1H), 9.62-10.48 (m, 2H).

Example 28

Step 1: Compound 18-1 (300 mg, 1.72 mmol) was dissolved in acetonitrile (5 mL) at 25 °C, 6-1 (414.41 mg, 1.72 mmol) and N,N-diisopropylethylamine ( 445.70 mg, 3.45 mmol, 600.67 μL), stirred at 0 °C for 1 hour, TLC showed that the raw material was consumed, 100 mL of saturated ammonium chloride solution was added to the reaction solution, and extracted with ethyl acetate (100 mL×3), the combined The organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product, which was subjected to column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0 ~4/1) was purified to obtain compound 28-1.

Step 2: Compound 28-1 (0.6 g, 1.59 mmol) was dissolved in dioxane (10 mL), 1-4 (397.15 mg, 1.75 mmol), cesium carbonate (1.03 g, 3.18 mmol) and [( 2-Di-cyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino- 1,1'-biphenyl)]palladium(II) methanesulfonate (143.94 mg, 158.79 μmol) was replaced with nitrogen three times and then heated to 100° C. and stirred for 4 hours under nitrogen protection. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-5/1) to obtain compound 28-2. MS _ESI calculated for _C28H44N8O3Si [ _M +H] ⁺ ₅₆₉ , found 569. ¹ H NMR (400MHz, MeOD)δ1.49-1.51(s,9H), 1.58-1.61(s,9H), 1.62-1.65(s,2H), 1.88-2.11(m,8H), 2.38-2.42( s,3H),2.46-2.54(m,4H),4.24-4.28(m,2H),4.39-4.53(m,1H),5.67(m,1H),6.35-6.44(m,1H),6.65- 6.71(m, 1H), 7.49-7.57(s, 1H).

Step 3: Compound 28-2 (500 mg, 879.07 μmol) was dissolved in DMF (10 mL) at 25°C, followed by adding water (0.5 mL), triethylamine (363.50 mg, 3.59 mmol, 0.5 mL) and selenium Powder (213.55 mg, 2.64 mmol), replaced with carbon monoxide three times and reacted at 90 ° C under 15 psi pressure for 1.5 hours, 50 mL of ammonium chloride aqueous solution was added to the reaction solution, and extracted with dichloromethane (50 mL × 3), the combined organic The phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 28-3.

Step 4: Compound 28-3 (600 mg, 923.42 μmol) was dissolved in ethanol (5 mL) at 25° C., 6-5 (85.44 mg, 923.42 μmol) was added, and the mixture was stirred at 80° C. for 1 hour. The reaction solution was directly concentrated under reduced pressure to obtain crude product 28-4. MS ESI _calcd for _{C31H48N8O3SeSi} [ _M +H] ⁺ ₆₈₉ , found 689.

Step 5: Compound 28-4 (600 mg, 872.34 μmol,) was dissolved in an ethyl acetate solution of hydrochloric acid (4 M, 5 mL) and stirred at 25° C. for 2 hours. The reaction solution was directly filtered to obtain the crude hydrochloride of compound 28-5 as the filter cake. MS _ESI calculated for _C20H26N8Se [ _M +H] ⁺ 459, found 459.

Step 6: The crude hydrochloride of 28-5 (100 mg, 202.47 μmol) was dissolved in methanol (2 mL), N,N-diisopropylethylamine (52.34 mg, 404.95 μmol, 70.53 μL) was added, and the mixture was After stirring at 25°C for 10 minutes, compound acrylonitrile (1-10) (10.74 mg, 202.47 μmol, 13.43 μL) was added, and the mixture was stirred at 25°C for 16 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 17%-47% , 10 min) to obtain the trifluoroacetate salt of compound 28-6. MS ESI calculated for C ₂₃ H ₂₉ N ₉ Se[M+H] ⁺ 512, found 512, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.84-2.09 (m, 4H), 2.11-2.29 (m, 4H), 2.32-2.52(m, 5H), 2.56-2.60(s, 3H), 3.08-3.13(m, 2H), 3.77-3.82(m, 2H), 3.85-3.89(m, 2H), 5.08- 5.21(m, 1H), 5.88-5.99(s, 1H), 6.78(s, 1H), 8.03-8.18(s, 1H).

Example 29

Step 1: Compound 29-1 (0.5 g, 4.50 mmol) was dissolved in DMF (10 mL) at 25° C., water (1 mL) and selenium powder (728.58 mg, 9.00 mmol) were added successively, and the solution was replaced with carbon monoxide three times. Under the pressure of 50 psi, the reaction was carried out at 80 °C for 16 hours. After TLC monitoring the consumption of the raw materials, 50 mL of ammonium chloride aqueous solution was added to the reaction solution, and extracted with dichloromethane (50 mL × 3), and the combined organic phase was then saturated brine. (50 mL) was washed, and finally the organic phase was dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0～1/1) to obtain Compound 29-2. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 1.69-1.84 (m, 5H), 3.81-3.93 (m, 4H).

Step 2: Compound 29-2 (0.4 g, 1.49 mmol) was added to methanol (10 mL), compound 8-4 (285.76 mg, 1.49 mmol) was added, and the reaction was carried out at 20° C. for 1 hour. TLC showed that the starting material was completely consumed. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-5/1) to obtain compound 29-3. MS ESI _calcd for _{C13H12Cl2N2OSe} [M ₊ H] ₊ 363, found 363 ^.

Step 3: Compound 29-3 (350 mg, 966.55 μmol), Compound 1-2 (262.49 mg, 1.16 mmol), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (120.37 mg, 193.31 μmol), palladium acetate (21.70 mg, 96.65 μmol), and cesium carbonate (629.84 mg, 1.93 mmol) were added to dioxane (5 mL) and reacted at 80° C. for 3 hours under nitrogen atmosphere. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-2/1) to obtain compound 29-4. MS ESI calculated: _{C25H33ClN4O3Se} [ _{M +} H] ⁺ ₅₅₃ , found 553.

Step 4: Compound 29-4 (250 mg, 452.93 μmol) was dissolved in dioxane (5 mL), compound 8-9 (98.27 mg, 498.22 μmol), cesium carbonate (295.14 mg, 905.85 μmol) and [( 2-Di-cyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino- 1,1'-biphenyl)]palladium(II) methanesulfonate (41.06 mg, 45.29 μmol) was replaced with nitrogen three times and then heated to 90° C. and stirred for 4 hours under nitrogen protection. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-10/1) to obtain compound 29-5. MS _ESI calculated for _C34H47N7O5Se [M+ _H ] ⁺ ₇₁₄ , found 714.

Step 5: Compound 29-5 (200 mg, 280.61 μmol) was dissolved in an ethyl acetate solution of hydrochloric acid (4 M, 2 mL) and stirred at 25° C. for 2 hours. The reaction solution was directly filtered to obtain the crude hydrochloride of compound 29-6 as the filter cake. MS ESI _calcd for _C24H31N7OSe [ _M +H] ⁺ 514, found 514.

Step 6: The crude hydrochloride of 29-6 (150 mg, 273.24 μmol) was dissolved in methanol (1 mL), N,N-diisopropylethylamine (70.63 mg, 546.48 μmol, 95.19 μL) was added, and the mixture was After stirring at 25°C for 10 minutes, compound acrylonitrile (1-10) (29.00 mg, 546.48 μmol, 36.25 μL) was added, and the mixture was stirred at 25°C for 16 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 20%-40% , 7 min) to obtain the trifluoroacetate salt of compound 29-7. MS _{ESI calcd} for _C27H34N8OSe [M+H] ⁺ 567, found 567. ¹ H NMR (400MHz, CD ₃ OD) δ 1.85-1.96 (m, 2H), 2.07-2.18 (m, 4H), 2.26-2.50 (m, 9H), 3.08-3.13 (t, 2H), 3.33- 3.39(m, 1H), 3.47-3.56(t, 2H), 3.58-3.65(t, 2H), 4.02-4.08(d, 2H), 4.18-4.29(m, 3H), 5.81-5.89(s, 1H ), 6.79-6.83(s, 1H), 6.89-6.92(s, 1H), 8.84(s, 1H).

Example 30

Step 1: Compound 30-1 (0.5 g, 8.76 mmol, 591.02 μL) was added to dichloromethane (5 mL) at 0°C, followed by triethylamine (2.66 g, 26.27 mmol, 3.66 mL) and 30-2 (1.11 g, 10.51 mmol, 773.00 μL), reacted at 20° C. for 2 hours. TLC showed that the starting material was completely consumed. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried with anhydrous sodium sulfate, filtered and reduced in pressure Concentration gave compound 30-3.

Step 2: Elemental selenium (1.58g, 19.49mmol) was added to ethanol (30mL), sodium borohydride (737.26mg, 19.49mmol) was added in batches at 0-5°C, after the addition, the reaction was performed at 20°C for 1 hour, and 30- 3 (0.8 g, 9.74 mmol), then pyridine hydrochloride (4.50 g, 38.98 mmol) was slowly added, and the mixture was reacted at 80° C. for 1 hour. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with dichloromethane (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried with anhydrous sodium sulfate, filtered and reduced in pressure Concentration gave a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether:ethyl acetate=1/1 to dichloromethane:methanol=10:1) to obtain compound 30-4. ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 1.96-2.21 (m, 2H), 3.73-4.07 (m, 4H), 7.26-7.84 (m, 2H).

Step 3: Compound 30-4 (0.45 g, 1.67 mmol) was added to methanol (2 mL), Compound 8-4 (272.89 mg, 1.67 mmol) and sodium fluoride (35.13 mg, 836.67 μmol) were added, and the reaction was carried out at 20°C for 0.5 hour . 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentrated to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-5/1) to obtain compound 30-5. MS ESI _calcd for _C11H9Cl2N3Se [ _{M +} H] ⁺ 334, found 334.

Step 4: Compound 30-5 (600 mg, 1.80 mmol), Compound 1-2 (407.68 mg, 1.80 mmol), (±)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (224.34 mg, 360.28 μmol), palladium acetate (40.44 mg, 180.14 μmol), and cesium carbonate (1.17 g, 3.60 mmol) were added to dioxane (4 mL) and reacted at 80° C. for 3 hours under nitrogen atmosphere. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-5/1) to obtain compound 30-6. MS _{ESI calcd} for _{C23H30ClN5O2Se} [M ₊ H] ⁺ 524, found 524.

Step 5: Compound 30-6 (400 mg, 764.92 μmol) was dissolved in dioxane (4 mL), compound 8-9 (165.96 mg, 841.42 μmol), cesium carbonate (498.45 mg, 1.53 mmol) and [( 2-Di-cyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino- 1,1'-biphenyl)]palladium(II) methanesulfonate methanesulfonate (69.34 mg, 76.49 μmol), replaced with nitrogen three times, then heated to 100° C., and stirred for 4 hours under nitrogen protection. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=5/1-1/1) to obtain compound 30-7. MS ESI calculated: C ₃₂ H ₄₄ N ₈ O ₄ Se[M+H] ⁺ 685, found 685.

Step 6: Compound 30-7 (350 mg, 397.31 μmol) was dissolved in an ethyl acetate solution of hydrochloric acid (4 M, 4 mL) and stirred at 25° C. for 16 hours. The reaction solution was directly concentrated under reduced pressure to obtain the crude hydrochloride of compound 30-8. MS _{ESI calcd} for _C22H28N8Se [M+H] ⁺ 485, found 485.

Step 7: The crude hydrochloride of 30-8 (250 mg, 480.83 μmol) was dissolved in methanol (5 mL), N,N-diisopropylethylamine (186.43 mg, 1.44 mmol, 251.26 μL) was added, and the mixture was After stirring at 25°C for 10 minutes, compound acrylonitrile (1-10) (51.03 mg, 961.66 μmol, 63.79 μL) was added, and the mixture was stirred at 25°C for 16 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 15%-45% , 10 minutes) to obtain the trifluoroacetate salt of compound 30-9. MS ESI calculated for C ₂₅ H ₃₁ N ₉ Se[M+H] ⁺ 538, found 538, ¹ H NMR (400 MHz, CD ₃ OD) δ 2.08-2.17 (s, 2H), 2.30-2.48 (m, 9H), 2.50-2.58(m, 2H), 3.09-3.13(s, 2H), 3.46-3.56(m, 2H), 4.14-4.26(m, 7H), 5.84(s, 1H), 6.66(s, 1H), 6.71 (s, 1H), 7.96 (s, 1H).

Example 31

Step 1: Compound 18-1 (360 mg, 2.07 mmol) was dissolved in acetonitrile (5 mL) at 0 °C, 6-1 (497.3 mg, 2.07 mmol) and N,N-diisopropylethylamine ( 534.8 mg, 4.14 mmol, 720.80 μL), stirred at 0 °C for 1 hour, 100 mL of saturated ammonium chloride solution was added to the reaction solution, and extracted with ethyl acetate (100 mL×3), the combined organic phase was then saturated brine (100 mL) was washed, and finally the organic phase was dried with anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0～2/1) to obtain Compound 31-1.

Step 2: Compound 33-1 (0.7 g, 1.85 mmol) was dissolved in tetrahydrofuran (10 mL) at 0 °C, sodium hydride (110 mg, 2.75 mmol, 60% purity) was added, stirred at 0 °C for 0.5 hours, and then Iodomethane (525.8 mg, 3.71 mmol, 230.6 μL, ) was slowly added to the system, and then the whole system was stirred at 35° C. for 3 hours. 100 mL of saturated ammonium chloride solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried with anhydrous sodium sulfate, filtered and reduced Concentrate under pressure to obtain a crude product, which is purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-1/1) to obtain compound 31-2. ¹ H NMR (400 MHz, CDCl ₃ ) δ 0.76-0.84 (m, 2H), 1.14-1.25 (m, 2H), 1.41-1.45 (s, 9H), 1.56-1.90 (m, 6H), 2.68-2.84 (s, 3H), 4.25-4.35 (m, 1H), 4.37-4.47 (m, 1H), 5.75-6.07 (m, 1H), 6.46-6.69 (m, 1H).

Step 3: Compound 31-2 (600 mg, 1.53 mmol) was dissolved in dioxane (5 mL), compound 1-4 (382.9 mg, 1.68 mmol), cesium carbonate (997.6 mg, 3.06 mmol) and [( 2-Di-cyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino- 1,1'-biphenyl)] palladium(II) methanesulfonate (138.8 mg, 153.10 μmol) methanesulfonate (138.8 mg, 153.10 μmol) was replaced with nitrogen three times and then heated to 100° C., and stirred for 4 hours under nitrogen protection. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (100 mL×3), the combined organic phase was washed with saturated brine (100 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure Concentration to obtain a crude product, which was purified by column chromatography (SiO ₂ , petroleum ether: ethyl acetate=1/0-1/1) to obtain compound 31-3. MS ESI calculated for _{C29H46N8O3Si [ M} +H] ⁺ ₅₈₃ , found 583.

Step 4: At 25°C, compound 31-3 (250 mg, 428.9 μmol, ) was dissolved in DMF (5 mL), followed by adding water (0.3 mL), triethylamine (218.1 mg, 2.16 mmol, 0.3 mL) and Selenium powder (104.2 mg, 1.29 mmol) was replaced with carbon monoxide three times and reacted at 90 ° C under 15 psi pressure for 2 hours, 50 mL of ammonium chloride aqueous solution was added to the reaction solution, and extracted with dichloromethane (50 mL × 3), the combined The organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 31-4.

Step 5: Compound 31-4 (300 mg, 451.95 μmol) was dissolved in ethanol (5 mL) at 25° C., 6-5 (50.2 mg, 542.34 μmol) was added, and the mixture was stirred at 80° C. for 3 hours. The reaction solution was directly concentrated under reduced pressure to obtain crude product 31-5. MS _ESI calculated for _{C32H50N8O3SeSi} [ _M +H] ⁺ ₇₀₃ , found 703.

Step 6: Compound 31-5 (0.3 g, 427.45 μmol) was dissolved in methanolic hydrochloric acid solution (4 M, 4 mL) and stirred at 30° C. for 1 hour. LCMS showed that the reaction of the starting materials was basically complete, and the main peak was the product peak. The reaction solution was directly concentrated under reduced pressure to obtain the crude hydrochloride of compound 31-6. MS ESI _calcd for _C21H28N8Se [M+H] ⁺ ₄₇₃ , found 473.

Step 7: The crude hydrochloride of 31-6 (200 mg, 393.76 μmol) was dissolved in methanol (2 mL), N,N-diisopropylethylamine (152.7 mg, 1.18 mmol, 205.8 μL) was added, and the mixture was After stirring at 25°C for 10 minutes, acrylonitrile (1-10) (41.8 mg, 787.52 μmol, 52.2 μL,) was added, and the mixture was stirred at 25°C for 16 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 15%-45% , 7 min) to obtain the trifluoroacetate salt of compound 31-7. MS ESI calculated for C ₂₄ H ₃₁ N ₉ Se[M+H] ⁺ 526, found 526, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.87-2.15 (m, 6H), 2.26-2.46 (m, 5H), 2.46-2.62(m, 5H), 3.10-3.15(t, 2H), 3.16-3.23(s, 3H), 3.78-3.84(t, 2H), 3.87-3.94(m, 2H), 5.97- 6.01(s, 1H), 6.02-6.21(m, 1H), 6.99-7.10(s, 1H), 8.12(s, 1H).

Example 32

Step 1: Compound 18-1 (0.6 g, 3.45 mmol) was dissolved in tetrahydrofuran (15 mL) at 20°C, and diisopropylethylamine (891.37 mg, 6.90 mmol) was added. Then 1-2 (780.45 mg, 3.45 mmol) was added dropwise at 0°C, reacted at 0°C for 2 hours, and then at 20°C for 16 hours. The reaction solution was diluted with 15 mL of water and extracted with 120 mL (40 mL×3) of ethyl acetate. The combined organic layers were washed with 40 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether: ethyl acetate= 10/1 to 1/1) to give compound 32-2. MS _ESI calculated: _C17H22ClN5O2 [M ₊ H] ⁺ ₃₆₄ , found 364.

Step 2: Compound 32-2 (740 mg, 1.98 mmol) was dissolved in DMF (15 mL), and sodium hydride (103.00 mg, 2.58 mmol) was added in batches at 0°C. After the addition, the reaction was carried out at this temperature for 30 minutes, and then 0 Iodomethane (309.29 mg, 2.18 mmol) was added dropwise at °C, and the mixture was reacted at 15 °C for 2 hours. 10 mL of saturated aqueous ammonium chloride solution was added, then 50 mL of water was added, and extracted with 40 mL of ethyl acetate (20 mL×2). The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, using petroleum ether:ethyl acetate =10mL: 10mL was beaten at room temperature to obtain compound 32-3. MS _ESI calculated: _C18H24ClN5O2 [ _{M +} H] ⁺ 378, found 378.

Step 3: Compound 32-3 (0.6 g, 1.59 mmol), 8-9 (344.50 mg, 1.75 mmol) were added to dioxane (15 mL) followed by [(2-di-cyclohexylphosphino-3 ,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2-(2'-amino-1,1'-biphenyl)] Palladium(II) methanesulfonate methanesulfonate (143.94 mg, 158.79 μmol), cesium carbonate (1.03 g, 3.18 mmol) were reacted at 80° C. for 3 hours under nitrogen atmosphere. The reaction solution was diluted with 50 mL of water, and extracted with 40 mL (20 mL×2) of ethyl acetate. The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 5/1 to 1/1) purification to give compound 32-4. MS _ESI calculated: _C27H38N8O4 [M ₊ H] ⁺ ₅₃₉ , found 539.

Step 4: DMF (5mL) and water (0.5mL) were added to the single-neck flask, followed by compound 32-4 (350mg, 630.29μmol), selenium powder (153.12mg, 1.89mmol) and triethylamine (352.59mg, 3.48mmol) ), replaced 3 times with carbon monoxide, and reacted at 90° C. for 1 hour under a carbon monoxide (15 psi) atmosphere. The reaction solution was diluted with 50 mL of water and extracted with 120 mL (40 mL×3) of ethyl acetate. The combined organic layers were washed with brine 50 mL, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 32-5. MS _ESI calculated: _C27H40N8O4Se [M ₊ H] ⁺ ₆₂₁ , found 621.

Step 5: Compound 32-5 (400 mg, 645.56 μmol) was dissolved in ethanol (10 mL) and chloroacetone (0.26 g, 2.81 mmol) was added. The reaction was carried out at 80°C for 2 hours. The reaction solution was concentrated to dryness, then Boc acid anhydride (169.07 mg, 774.67 μmol), triethylamine (195.97 mg, 1.94 mmol) and tetrahydrofuran (6 mL) were added, heated to 50° C. to react for 3 hours, concentrated to dryness under reduced pressure, and passed through a silica gel plate ( Petroleum ether:ethyl acetate=1:2) was purified to obtain compound 32-6. MS _ESI calculated: _C30H42N8O4Se [M ₊ H] ⁺ ₆₅₉ , found 659.

Step 6: Compound 32-6 (50 mg, 76.03 μmol) was added to a single-neck flask, ethyl acetate hydrochloride (4 M, 3 mL) was added, and the reaction was carried out at 50° C. for 1 hour. The reaction solution was concentrated under reduced pressure to obtain compound 32-7 hydrochloride. MS ESI calculated: _C20H26N8Se [ _M +H] ₊₄₅₉ , found 459.

Step 7: Compound 32-7 hydrochloride (50 mg, 101.24 μmol) was dissolved in methanol (2 mL), diisopropylethylamine (65.42 mg, 506.20 μmol) and acrylonitrile (1-10) (10.74 mg were added) , 202.48 μmol), and reacted at 15 °C for 2 hours. The reaction solution was added with 30 mL of water, extracted with 40 mL (20 mL×2) of ethyl acetate, the combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography (column type: Phenomenex Luna C18 75×30 mm×3 μm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; acetonitrile %: 15%-45%, 7 minutes) to obtain the trifluoroacetic acid salt of compound 32-8. MS _ESI calculated: _C23H29N9Se [M+H] ⁺ ₅₁₂ , found 512. ¹ H NMR (400 MHz, CD ₃ OD) δ 2.01 (d, J=12.0 Hz, 2H), 2.26-2.37 (m, 5H), 2.37-2.51 (m, 4H), 2.58 (s, 3H), 3.13 (t, J＝7.2Hz, 2H), 3.18(s, 3H), 3.49(t, J＝6.8Hz, 2H), 4.27(s, 2H), 5.24-5.81(m, 1H), 6.03(s, 1H), 7.04(s, 1H), 8.09(s, 1H).

Example 33

Step 1: The hydrochloride of 32-7 (60 mg, 104.99 μmol) was dissolved in methylpyrrolidone (1 mL), triethylamine (53.1 mg, 524.9 μmol, 73.0 μL) was added, and the mixture was stirred at 20° C. for 30 minutes, Then, the reaction system was cooled to 0°C, compound 3-1 (20.86 mg, 115.49 μmol) was added, and the mixture was stirred at 20°C for 2 hours. 50 mL of ammonium chloride aqueous solution was added to the reaction solution, extracted with ethyl acetate (50 mL×3), the combined organic phase was washed with saturated brine (50 mL), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and reduced in pressure The crude product was obtained by concentration, and the crude product was prepared and separated by high performance liquid chromatography (column: Phenomenex Luna C18 75*30mm*3μm; mobile phase: [water (0.1%TFA)-ACN]; B(ACN)%: 30%-60% , 7 minutes) to obtain the trifluoroacetate salt of compound 33-1. MS ESI calculated for C ₂₄ H ₃₀ N ₁₀ O ₂ SSe[M+H] ⁺ 603, found 603, ¹ H NMR (400 MHz, CD ₃ OD) δ 1.75-1.86 (m, 2H), 1.96-2.10 ( m, 2H), 2.11-2.25(m, 4H), 2.36(s, 3H), 2.61(s, 3H), 3.15-3.24(s, 3H), 3.64-3.74(m, 1H), 4.03-4.11( t,2H),4.15-4.24(t,2H),4.28-4.38(m,2H),5.45-5.70(m,1H),5.87-6.04(s,1H),6.92-7.12(s,1H), 8.15(s, 1H).

Example 34

Step 1: At 20°C, diisopropylethylamine (6.24g, 48.31mmol) and compound 20-1 (5g, 24.15mmol) were added to tetrahydrofuran (80mL), compound 1-2 (5.47g, 24.15mmol) ) was dissolved in tetrahydrofuran (20 mL), dropped into the former system at 0°C, and reacted at 20°C for 16 hours. The reaction solution was diluted with 200 mL of water, extracted with 200 mL of ethyl acetate (100 mL×2), the combined organic layers were washed with 100 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 10/1 to 1/1) to give compound 34-1. MS ESI calculated: _C18H25ClN4O4 [M ₊ H] _{+ 397} , found 397 ^.

Step 2: Compound 34-1 (5.23 g, 13.18 mmol) was dissolved in tetrahydrofuran (80 mL), and sodium hydride (632.49 mg, 15.81 mmol) was added in portions at 0°C. After the addition, the reaction was carried out at 0°C for 30 minutes. Iodomethane (2.06 g, 14.50 mmol) was dissolved in tetrahydrofuran (20 mL), added dropwise to the above reaction system at 0°C, and then reacted at 15°C for 16 hours. The reaction solution was quenched by adding 20 mL of ammonium chloride solution, diluted with 150 mL of water, extracted with 160 mL of ethyl acetate (80 mL×2), the combined organic layers were washed with 80 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (Petroleum ether/ethyl acetate=4/1 to 3/1) to obtain compound 34-2. MS ESI calculated: _C19H27ClN4O4 [M ₊ H] _{+ 411} , found 411 ^.

Step 3: Compound 34-2 (2.4 g, 5.84 mmol) was dissolved in tetrahydrofuran (50 mL), methylmagnesium bromide ether solution (3M, 3.11 mL,) was added dropwise at 0°C, and the reaction was carried out at -78°C for 1 hour. 5 mL of saturated ammonium chloride solution was added to quench the reaction, the reaction solution was diluted with 60 mL of water, extracted with 60 mL of ethyl acetate (30 mL × 2), the combined organic layers were washed with 30 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure. Silica gel column purification (petroleum ether/ethyl acetate=3:1) gave compound 34-3. ¹ H NMR (400 MHz, CDCl ₃ ) δ 1.50 (s, 9H), 1.61 (dd, J=12.4, 3.2 Hz, 2H), 1.85 (d, J=6.8 Hz, 4H), 2.05 (d, J=6.4 Hz, 2H), 2.62(s, 3H), 2.86(s, 3H), 4.32(s, 2H), 5.30(s, 1H), 6.92(s, 1H)

Step 4: Compound 34-3 (800 mg, 2.03 mmol) was dissolved in dichloromethane (20 mL), triethylamine (1.23 g, 12.16 mmol) was added, and tert-butyldimethylsilyl trifluoromethyl was added dropwise Sulfonate (1.61 g, 6.08 mmol, ) was reacted at 15°C for 16 hours. The reaction solution was diluted with dichloromethane 60 mL, and was diluted with saturated aqueous sodium bicarbonate solution 60 mL (30 mL×2), water (20 mL) and brine 30 mL, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 34-4. MS ESI calculated: _{C25H41ClN4O3Si} [ _{M +} H] ⁺ ₅₀₉ , found 509.

Step 5: Compound 34-4 (1.15 g, 2.03 mmol) was dissolved in tetrahydrofuran (30 mL) and water (5 mL), bromosuccinimide (721.57 mg, 4.05 mmol) was added, and the reaction was carried out at 15° C. for 2 hours. The reaction solution was diluted with 100 mL of water, extracted with 100 mL of ethyl acetate (50 mL×2), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain compound 34-5. MS ESI calculated: _{C19H26BrClN4O3 [} M ₊ H] ⁺ ₄₇₃ , found 473.

Step 6: Compound 34-5 (800 mg, 2.14 mmol) was dissolved in methanol (25 mL), methylselenamide (12-2) (261.25 mg, 2.14 mmol) was added at 0-5 °C, and the reaction was carried out at 15 °C for 2 hours . Then BOC acid anhydride (560.70 mg, 2.57 mmol), 2,6-lutidine (26.16 mg, 214.09 μmol) and triethylamine (866.55 mg, 8.56 mmol) were added, and the reaction was carried out at 15° C. for 2 hours. The reaction solution was diluted with 100 mL of water, extracted with 90 mL of ethyl acetate (30 mL×3), the combined organic layers were washed with 50 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 5/1 to 3/1) to give compound 34-6. MS ESI calculated: _{C21H28ClN5O2Se} [ _{M +} H] ⁺ ₄₉₈ , found 498.

Step 7: Compound 34-6 (120 mg, 241.50 μmol), tert-butyl 5-amino-3-methyl-1H-pyrazole-1-carboxylate (57.16 mg, 289.80 μmol) was added to dioxane (4 mL) Add [(2-di-cyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)-2- (2'-Amino-1,1'-biphenyl)]palladium(II) mesylate (21.89 mg, 24.15 μmol) mesylate and cesium carbonate (157.37 mg, 483.00 μmol) at 80°C under nitrogen atmosphere The reaction was carried out for 3 hours. The reaction solution was diluted with 50 mL of water, and extracted with 40 mL (20 mL×2) of ethyl acetate. The combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by silica gel column (petroleum ether/ethyl acetate= 5/1 to 1/1) to give compound 34-7. MS _ESI calculated: _C30H42N8O4Se [M ₊ H] ⁺ ₆₅₉ , found 659.

Step 8: Compound 34-7 (60 mg, 91.23 μmol) was added to hydrochloric acid methanol (4 M, 3 mL), and the reaction was carried out at 40° C. for 1 hour. The reaction solution was concentrated under reduced pressure to obtain the hydrochloride of compound 34-8. MS ESI calculated: C ₂₀ H ₂₆ N ₈ Se[M+H] ⁺ 459, found 459.

Step 9: The hydrochloride salt of compound 34-8 (55 mg, 120.24 μmol) was dissolved in methanol (2 mL), diisopropylethylamine (62.16 mg, 480.94 μmol) and acrylonitrile (1-10) (12.76 μg) were added mg, 240.47 μmol), reacted at 15°C for 2 hours. The reaction solution was added with 30 mL of water, extracted with 40 mL (20 mL×2) of ethyl acetate, the combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography (column type: 3- 100 C18 ultra 150*50mm*3μm; mobile phase: [water (0.225% formic acid)-acetonitrile]; acetonitrile %: 5%-35%, 10 minutes) to obtain the formate salt of compound 34-9. MS _ESI calculated: _C23H29N9Se [M+H] ₊₅₁₂ , found 512. ¹ HNMR (400MHz, CD ₃ OD) δ 1.48-1.65(m, 2H), 1.83-1.91(m, 2H), 1.94-2.03(m, 2H), 2.05-2.14(m, 2H), 2.29(s) ,3H),2.61-2.69(m,2H),2.72-2.77(m,2H),2.81(s,3H),2.99(s,3H),3.46(s,2H),5.00-5.60(m,1H ), 6.22(s, 1H), 6.81(s, 1H), 8.81(s, 1H).

Example 35

Step 1: The hydrochloride salt of compound 34-8 (110 mg, 222.72 μmol) was dissolved in tetrahydrofuran (2 mL), triethylamine (112.68 mg, 1.11 mmol, ) and compound 3-1 (40.23 mg, 222.72 μmol) were added, The reaction was carried out at 15°C for 1 hour. The reaction solution was added with 30 mL of water, extracted with 40 mL (20 mL×2) of ethyl acetate, the combined organic layers were washed with 20 mL of brine, dried over sodium sulfate, filtered and concentrated under reduced pressure, and purified by high performance liquid chromatography (column type: Phenomenex luna C18 150*25mm*10μm; mobile phase: [water (0.225% formic acid)-acetonitrile]; acetonitrile %: 16%-46%, 10 minutes) to obtain the formate salt of compound 35-1. MS ESI calculated: C ₂₄ H ₃₀ N ₁₀ O ₂ SSe[M+H] ⁺ 603, found 603. ¹ HNMR (400MHz, CD ₃ OD) δ 1.68 (d, J=10.0 Hz, 2H), 1.93 (d, J=7.6 Hz, 2H), 1.99-2.15 (m, 4H), 2.28 (s, 3H) ,2.80(s,3H),2.94(s,3H),3.65(s,1H),4.01(t,J＝7.2Hz,2H),4.09-4.17(m,2H),4.22(s,2H), 5.18-5.66(m, 1H), 6.14(s, 1H), 6.77(s, 1H), 8.82(s, 1H).

Example 36

Step 1: Compound 5-1 (7.69 mg, 40.62 μmol) was dissolved in DMF (0.3 mL) at 20°C, 1-hydroxybenzotriazole (10.98 mg, 81.24 μmol) and 1-(3- Dimethylaminopropyl)-3-acetaldehyde hydrochloride (15.57 mg, 81.24 μmol), and the mixture was stirred at 20° C. for 0.5 hr. A solution of compound 18-6 hydrochloride (30 mg, 44.68 μmol) and diisopropylethylamine (15.75 mg, 121.85 umol) in dimethylformamide (0.3 mL) was added to the mixture, and stirred at 20° C. for 16 Hour. LC-MS showed that 33% of the starting material remained, and the reaction solution was supplemented with 5-1 (3.84 mg, 20.31 μmol), followed by stirring at 20° C. for 2 hours. 10 mL of water was added to the reaction solution, extracted with ethyl acetate (20 mL×3), the combined organic phase was washed with saturated brine (5 mL×3), and finally the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, The crude 36-1 was obtained. MS _ESI calculated: _C27H37N9O3Se [ _M +H] ⁺ ₆₁₆ , found 616.

Step 2: Compound 36-1 (60 mg, 97.62 μmol) was dissolved in dichloromethane (3 mL) at 20°C, trifluoroacetic acid (1 mL) was added, and the mixture was stirred at 20°C for 1 hour. LC-MS showed that the starting material was consumed and the main peak was the product peak. The reaction solution was concentrated and spin-dried to obtain a crude product. The crude product was separated by high performance liquid chromatography (column: Phenomenex Synergi C18 150*25mm*10μm; mobile phase: [water (0.1% TFA)-ACN]; ACN%: 13%-43%, 10 minutes) to obtain compound 36 -2's trifluoroacetate. MS ESI calculated: C ₂₂ H ₂₉ N ₉ OSe [M+H] ⁺ 516, found 516. ¹ H NMR(400MHz,DMSO-d6)δ1.36-1.52(m,2H),1.78-1.88(m,2H),1.91(m,2H),2.07(m,2H),2.21(s,3H) ,2.30(s,3H),2.41(s,3H),3.16-3.27(m,2H),3.39m,1H),4.31(m,1H),4.53(m,2H),6.32-6.49(m, 1H), 6.51(s, 1H), 7.24(m, 1H), 7.86(s, 1H), 8.88-9.18(m, 1H), 11.59-12.04(m, 1H).

biological test

Experimental example 1:

Main reagents and consumables

experimental method

JAK1, 2, 3 and TYK2 were used in this experiment

method for activity detection. In the assay plate, the enzyme, ULight-labeled polypeptide substrate, ATP, and detection compound are mixed and the reaction incubated. After the reaction, EDTA was added to stop the reaction, and Eu-labeled antibody was added at the same time. exist

In kinase assays, the binding of europium-labeled anti-phosphorylated substrate antibodies to phosphorylated ULight-labeled substrates enables donor and acceptor molecules to approach each other. After irradiation with 320nm wavelength light, the kinase reacts, the energy of the europium donor is transferred to the ULight acceptor dye, and the 665nm wavelength light is generated. The emission intensity of light is proportional to the phosphorylation level of the ULight matrix.

The final test concentration of the compound: the final test concentration of the test compound is from 1 μM to 0.017 nM, 3-fold serial dilution, 11 concentrations. The content of DMSO in the detection reaction was 1%.

Kinase assay: Preparation of buffers including: 50 mM HEPES (pH 7.5), 0.01% Brij-35, 10 mM MgCl ₂ , 1 mM EDTA, 1 mM DTT.

JAK1 enzymatic reaction:

In buffer, 2nM JAK1 and 50nM substrate were pre-incubated for 15 minutes with a mixture of pre-diluted compounds at various concentrations. The reaction was started by adding 38 μM ATP and incubated at room temperature for 90 minutes. After the reaction was completed, antibody detection was added, and after incubation at room temperature for 60 minutes, Evnvision detection was performed, and data was collected.

JAK2 enzymatic reaction:

In buffer, 0.03 nM JAK2 and 50 nM substrate were pre-incubated for 15 minutes with a mixture of pre-diluted compounds at various concentrations. The reaction was started by adding 12 μM ATP and incubated at room temperature for 90 minutes. After the reaction was completed, antibody detection was added, and after incubation at room temperature for 60 minutes, Evnvision detection was performed, and data was collected.

data analysis:

According to % inhibition vs. log [compound concentration], XLfit5 software mode205 was used to perform data analysis and plot to obtain IC ₅₀ data. The results are summarized in Table 1.

Table 1: Summary of Kinase Activity of Test Compounds

Conclusion: The test compound of the present invention exhibits a good inhibitory effect on the in vitro activity test of two subtypes of kinases, JAK1 and JAK2.

Experimental example 2

Main reagents and instruments

Main reagent consumables

name	supplier	article number
THP1	ATCC	TIB-202
HT29	ATCC	HTB-38
Interleukin 6 (IL-6)	Absin	abs00808
Interleukin 13 (IL-13)	Peprotech	200-13
pSTAT3 antibody	BD	562072
pSTAT6 antibody	BD	562078
cell fixative	BD	558049
cell rupture fluid	BD	558050

staining buffer	Biolegend	420201
1640 culture medium	Gibco	22400105
penicillin/streptomycin	Hyclone	SV30010
fetal bovine serum	Hyclone	SV30087.03
Ethylenediaminetetraacetic acid	Invitrogen	15575-038
96-hole round bottom plate	Corning	3799

instrument

Flow cytometer: Brand: BD; Model: Fortessa

Reagent configuration:

Complete medium: 1640 medium + 10% fetal bovine serum + 1% penicillin/streptomycin (all percentages are volume ratios)

Experimental procedure

a) Drug treatment and induction

(1) Pour off the HT29 cell culture medium, add 10 mM ethylenediaminetetraacetic acid, and incubate at 37°C for 5 min to digest the cells.

(2) HT29 cells and THP1 cells were centrifuged together at 320 g for 3 min.

(3) Count the cells, and then adjust the cell concentration to 7.5×10 ⁵ /mL with complete culture medium; 200 μL/well is inoculated into a 96-well round bottom plate.

(4) Add drugs to the cells, and the final concentrations are 5000.00nM, 1000.00nM, 200.00nM, 40.00nM, 8.00nM, 1.60nM, 0.32nM, 0.06nM. Incubate at 37°C for 30min (negative and positive control groups do not add drugs, add the same concentration of dimethyl sulfoxide).

(5) IL-13 (final concentration 6ng/mL) was added to HT29 cells and incubated at 37°C for 30min; IL-6 (final concentration 30ng/mL) was added to THP1 cells and incubated at 37°C for 15min (the negative control group did not add Cytokine stimulation, positive control group added the same concentration of cytokines).

b) Cell staining and flow cytometry detection

(1) Centrifuge the above HT29 and THP1 cells at 320 g for 3 min.

(2) Wash cells twice with staining buffer.

(3) 100 μL of cell fixative solution was added to each well, and the cells were fixed at 4°C for 15 min.

(4) Wash the cells twice with staining buffer.

(5) 100 μL of cell permeation solution was added to each well, and the membrane was ruptured at 4°C for 30 min.

(6) The pSTAT3 antibody or pSTAT6 antibody was added to the staining buffer at a ratio of 2:48, and 50 μL/well was added to THP1 or HT29 cells, respectively, and stained at 4°C for 30 min.

(7) Wash cells twice with staining buffer.

(8) The cells were resuspended in 150 μL of staining buffer, and the mean fluorescence intensity (MFI) of PE channel (pSTAT) was detected by flow cytometry.

c) Data processing

(1) The MFI corresponding to the sample was obtained by flow cytometry.

(2) Response rate (Response%)=100×(sample MFI-negative control group MFI)/(positive control group MFI-negative control group MFI)

(3) The response rate was brought into graphpad prism8 software, and the curve was fitted by the method of log(inhibitor) vs.response--Variable slope(four parameters) to obtain the median inhibitory concentration (IC ₅₀ ).

The results of the JAK inhibitory activity test in THP1 and HT29 cells are shown in Table 2.

Table 2: Summary of Cellular Activity of Test Compounds

Conclusion: The test compounds of the present invention exhibited good inhibitory activity in the in vitro activity test of cell (THP1 and HT29) functional experiments.

Experimental example 3

Test animals:

SD rats, 2 per group, male, Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.

Dosing:

There were 2 male SD rats at each time point, 4 time points in total, 8 rats in total; after an overnight fast, the rats were administered p.o. at a dose of 5 mg/kg and an administration volume of 5 mL/kg.

Sample Collection:

After administration, the rats were sacrificed by CO ₂ at 1, 3, 6, and 12 hours. About 200 μL of blood was collected from the jugular vein, placed in an EDTA-K2 test tube, and centrifuged at 4°C, 3,200 g for 10 min to obtain plasma. Store at ±10°C; take the small intestine and colon at each time point, squeeze out the contents of the colon and small intestine, rinse with normal saline, weigh and homogenize (the homogenate solution is methanol:15mM PBS=1:2), and the homogenate ratio is 1:4 (1g tissue 4ml homogenate), stored at -70±10℃.

Sample processing:

Plasma sample processing:

Protein precipitation: add 200 μL of acetonitrile containing internal standard to 20 μL of plasma sample, after mixing, centrifuge at 12,000 g at 4 °C, take 50 μL of the supernatant after treatment, add 50 μL of the supernatant to the 96-well plate, centrifuge at 3,220 g at 4 °C, and perform LC- MS/MS analysis.

Small intestine homogenate sample processing:

Add 400 μL of acetonitrile containing internal standard to 40 μL of small intestine homogenate sample, after mixing, centrifuge at 12,000 g at 4 °C, take 50 μL of the supernatant after treatment, add 50 μL of the supernatant to 96-well plate, centrifuge at 3220 g at 4 °C, and directly carry out LC-MS/ MS analysis.

Colon homogenate sample processing:

Add 400 μL of acetonitrile containing internal standard to 40 μL of colon homogenate sample, after mixing, centrifuge at 12,000 g at 4°C, take 50 μL of the supernatant after treatment, add 50 μL of supernatant to 96-well plate, centrifuge at 3,220 g at 4°C, and carry out LC-MS directly on the supernatant. /MS analysis.

The experimental results are as follows:

Conclusion: The compounds of the present invention show good drug exposure levels in the small intestine and colon of rats, and the small intestine/plasma and colon/plasma ratios of the compounds are high, showing good tissue selectivity.

Experimental example 4

Test animals:

C57BL/6J mice, 2 per group, male, Lingchang Biotechnology Co., Ltd.

Dosing:

There were 2 male C57BL/6J mice at each time point, 4 time points, 8 mice in total; after an overnight fast, they were administered p.o. at a dose of 3 mg/kg, and the administration volume was 3 mL/kg.

Sample Collection:

After administration, mice were sacrificed by CO ₂ at 0.5, 1, 5, and 12 hours. About 30 μL of blood was collected by saphenous vein or cardiac puncture, placed in an EDTA-K2 test tube, and centrifuged at 3,200 g for 10 min at 4°C to obtain plasma. Store at -60°C or below; take the small intestine and colon at each time point, squeeze out the contents of the colon and small intestine, rinse with normal saline, weigh and homogenize (the homogenate is methanol: 15mM PBS = 1:2), homogenize The ratio of slurry is 1:4 (1g tissue 4mL homogenate), and it is stored at -60℃ or below.

Sample processing:

Plasma sample processing:

Protein precipitation: 60 μL of acetonitrile containing internal standard was added to 3 μL of plasma sample, mixed and centrifuged at 12,000g at 4°C. After treatment, 50 μL of supernatant was added to a 96-well plate, centrifuged at 3,220g at 4°C, and the supernatant was directly subjected to LC-MS. /MS analysis.

Small intestine homogenate sample processing:

Add 400 μL of acetonitrile containing internal standard to 20 μL of small intestine homogenate sample, and after mixing, centrifuge at 3,220 g at 4 °C, take 50 μL of the supernatant after processing, add 50 μL of the supernatant to the 96-well plate, centrifuge at 3,220 g at 4 °C, and directly perform LC-MS on the supernatant. /MS analysis.

Colon homogenate sample processing:

Add 400 μL of acetonitrile containing internal standard to the 20 μL colon homogenate sample, and after mixing, centrifuge at 3220g at 4 °C, take 50 μL of the supernatant and add it to a 96-well plate, centrifuge at 3220 g at 4 °C, and then directly conduct LC-MS/ MS analysis.

The experimental results are shown in the following table:

Note: ND, below detection limit

Conclusion: The compounds of the present invention show good drug exposure levels in the small intestine and colon of mice, and the small intestine/plasma and colon/plasma ratios of the compounds are high, showing good tissue selectivity.

Experimental example 5

A mouse model of enteritis induced by oxazolone (OXA)

Test animals:

45 Balb/c mice, 8-10 weeks old, 18-20 g, female.

Experimental steps:

Randomly divided into 9 groups, blank control group, 5 mice, 100 microliters of acetone + olive oil (4:1) back sensitization for 5 days, rectal perfusion of 50% ethanol started on Day 0; the other 8 model groups, Five mice in each group were sensitized with 100 μl of 2% Oxazolone (diluted in acetone + olive oil) for 5 days, and rectal infusion of 150 μl of 1% Oxazolone solution was started on Day 0. The compounds to be tested were administered from Day-1 to the end of Day3, and Day4 was the end point of the experiment. The specific groups are shown in Table 3.

Table 3 Animal grouping and dosing schedule

Note: The solvent is 0.5% CMC-Na

data processing

Experiment period (In-life)

The body weight and Disease Activity Index (DAI) scores of the animals were recorded every day, which were used to evaluate the morbidity of each group of animals and the effect of the test compound on the disease. The DAI score consists of 3 parts, and the specific criteria refer to Table 4 below.

Table 4 DAI scoring criteria

score	% weight loss	Stool traits	Occult blood or bloody stools
0	0	normal	occult blood negative
1	1 to 5	soft stool	occult blood weakly positive
2	6 to 10	loose stool	occult blood positive
3	11～20	loose stool	a small amount of blood
4	>20	watery stool	a lot of blood

Description of the method for the determination of fecal occult blood

See feces or fresh blood in the anus, no longer detect occult blood. For the rest, the feces of mice with no obvious bloody stool were collected, and occult blood was measured. If the stool becomes darker and darker within 1-2 minutes, give 2 points. If there is no obvious color in the feces within 1-2 minutes or the color is very weak, and there is color behind, but the color depth is significantly lower than the color of the mouse feces of 2 points, give 1 point.

Experimental results

The obtained data were expressed as mean ± standard error, and statistical differences were analyzed. The results are shown in Table 5, Table 6 and Table 7.

Table 6 DAI score changes

Table 7 Colon length and weight changes

Conclusion: The compounds of the present invention can alleviate the OXA-induced weight loss, significantly improve the disease activity index (DAI) score and the end point colon weight-length ratio in the oxazolone (OXA)-induced mouse enteritis model, showing good performance. treatment effect.

Claims (18)

1. A compound of formula (II) or a pharmaceutically acceptable salt thereof,

   

   in,

   L ₁ is -(CH ₂ ) _m -, -C(=O)-, -S(=O) ₂ - or -CH ₂ -C(=O)-;

   T ₁ is N or CH;

   T ₂ is N or CH;

   _T3 is N or CH;

   R ₁ is H, CN, C _1-3 alkyl, -NH-C _1-3 alkyl, 4-6 membered heterocycloalkyl or 5-6 membered heteroaryl, wherein the C _1-3 alkyl , -NH-C _1-3 alkyl, 4-6 membered heterocycloalkyl and 5-6 membered heteroaryl, each independently optionally substituted with 1, 2 or 3 R _a ;

   _R2 is _R21 or

   

   D ₁ is O, NH or CH ₂ ;

   D ₂ is CH or N;

   E ₁ and E ₂ are each independently a single bond or CH ₂ ;

   R ₂₁ is H, CN, NH ₂ , C _1-3 alkyl, -NH-C _1-3 alkyl, -N(C _1-3 alkyl) ₂ , -C(=O)-NH-C _{1 -3} alkyl and -C(=O)-NH-C _3-6 cycloalkyl, wherein said C _1-3 alkyl, -NH-C _1-3 alkyl, -N(C _1-3 alkane base) ₂ , -C(=O)-NH-C _1-3 alkyl and -C(=O)-NH-C _3-6 cycloalkyl, respectively, independently optionally replaced by 1, 2 or 3 R _b replace;

   R ₃ is H or C _1-3 alkyl;

   R ₄ is H or C _1-3 alkyl;

   q is 1 or 2;

   m is 0, 1 or 2;

   n is 1 or 2;

   _Ra and _Rb are each independently F, Cl, Br, I, OH, _NH2 , CN or COOH;

   The "4-6-membered heterocycloalkyl" and "5-6-membered heteroaryl" each independently contain 1, 2 or 3 independently -O-, -NH-, -S-, -Se- or A heteroatom or heteroatom group of N.
2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ is H, CN, CH ₃ , -NH-CH ₃ ,

   

   

   wherein the CH ₃ , -NH-CH ₃ ,

   

   are each independently optionally substituted with 1, 2 or 3 R _a .
3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein R ₁ is H, CN, CF ₃ , -NH-CH ₃ ,

   

   
4. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

   

   for

   

   
5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₂₁ is H, CN, NH ₂ , CH ₃ ,

   

   

   wherein the CH ₃ ,

   

   Each independently is optionally substituted with 1 , 2 or 3 R _b .
6. The compound according to claim 5 or a pharmaceutically acceptable salt thereof, wherein R ₂ is H, CN, NH ₂ , CH ₃ ,

   

   
7. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₃ is H or CH ₃ .
8. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₄ is H or CH ₃ .
9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein L ₁ is -CH ₂ -, -(CH ₂ ) ₂ -, -C(=O)-, -S(=O) ₂ -or _-CH2 -C(=O)-.
10. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

    

    for

    

    
11. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

    

    for

    
12. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

    

    for

    

    
13. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is

    

    in,

    n, R ₁ , R ₂ , R ₃ , L ₁ , T ₃ , T ₂ and q are as defined in claim 1 .
14. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is

    

    in,

    R ₁ , R ₂ , R ₃ , L ₁ , T ₂ and q are as defined in claim 1 .
15. A compound of the formula or a pharmaceutically acceptable salt thereof,

    

    
16. Use of the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the treatment of JAK-related diseases.
17. Use of the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 15 in the manufacture of a medicament for treating JAK-related diseases limited to the intestinal tract.
18. The use according to claim 17, wherein the JAK-related disease limited to the intestine is inflammatory bowel disease.