WO2021180006A1

WO2021180006A1 - Vinyl-substituted pyridine compounds

Info

Publication number: WO2021180006A1
Application number: PCT/CN2021/079337
Authority: WO
Inventors: 陆剑宇; 贺辉君; 胡国平; 胡利红; 丁照中; 黎健; 陈曙辉
Original assignee: 南京明德新药研发有限公司
Priority date: 2020-03-10
Filing date: 2021-03-05
Publication date: 2021-09-16
Also published as: CN115244041A

Abstract

Provided are a class of vinyl-substituted pyridine compounds (I) and pharmaceutically acceptable salts thereof, and the use of the compounds and the pharmaceutically acceptable salts thereof in the preparation of drugs for treating related diseases.

Description

Vinyl substituted pyridine compounds

This application claims the following priority:

CN202010163709.9, application date March 10, 2020.

Technical field

The present invention relates to a class of vinyl-substituted pyridine compounds and pharmaceutically acceptable salts thereof, and the application of such compounds and pharmaceutically acceptable salts in the preparation of drugs for treating related diseases.

Background technique

Tyrosine kinases can be classified as growth factor receptor (e.g., Axl, Mer, EGFR, PDGFR, FGFR and erbB2) or non-receptor (e.g. c-src and bcr-abl) kinases. Receptor tyrosine kinases are composed of about 20 different subfamilies. Non-receptor tyrosine kinases constitute many subfamilies. These tyrosine kinases have a variety of biological activities. Receptor tyrosine kinase is a large enzyme that spans the cell membrane. It has an extracellular binding domain for growth factors, a transmembrane domain and an intracellular part as a kinase, which phosphorylates specific tyrosine residues in proteins and therefore affects Cell Proliferation. Abnormal or inappropriate protein kinase activity can lead to an increase in disease states associated with this abnormal kinase activity.

Disruption of VEGF receptor signaling is a very attractive therapeutic target in cancer, because angiogenesis is a prerequisite for the growth of all solid tumors. Angiogenesis is an important part of certain normal physiological processes (such as embryogenesis and wound healing), but abnormal angiogenesis leads to some pathological diseases, especially tumor growth. Recent studies have found that VEGF can directly inhibit tumor immune response, and anti-angiogenesis therapy can enhance tumor immunity through blood vessel normalization. (Fukumura et al. "Enhancing cancer immunotherapy using antiangiogenics: opportunities and challenges", Nat Rev Clin Oncol. 2018). VEGF-A (vascular endothelial growth factor A) is a key factor that promotes tumor neovascularization (angiogenesis). VEGF induces endothelial cell proliferation and migration through signal transduction through receptors, namely VEGFR-1 (Flt-1), VEGFR-2 (KDR) and VEGFR-3 (Flt-4). These signal transduction responses are heavily dependent on receptor dimerization and activation of intrinsic receptor tyrosine kinase (RTK) activity. The binding of VEGF as a disulfide bond-linked homodimer stimulates receptor dimerization and activation of the RTK domain. Kinase activity autophosphorylates cytoplasmic receptor tyrosine residues, which then serve as binding sites for molecules involved in the proliferation of the signal cascade. Although the three receptors may have multiple pathways elucidated, VEGFR-2 signaling is the most widely studied, and its mitogenic response is thought to involve ERK-1 and ERK-2 mitogen-activated protein kinases.

Axl is a member of the TAM receptor family, which also includes Mer and Tyro3. These three are activated by a common ligand, growth arrest-specific protein 6 (Gas6), and they usually play a role in embryonic development in cell survival, migration and differentiation. Axl overexpression and signal transduction are associated with a variety of malignant tumors in humans, such as colon cancer, breast cancer, glioma, thyroid cancer, gastric cancer, melanoma, lung cancer, and renal cell carcinoma.

The inhibition of Axl can more completely prevent the growth of cancer. The two play a complementary and overlapping role. Therapies targeting these two RTKs may be more effective than single-targeting agents. In addition, therapeutic antibodies that target Axl can block the Axl function of the tumor stroma. Combining the inhibition of Axl with the inhibition of VEGF receptors (VEGFRs), its additive effect suggests that blocking the function of Axl is one of the promotion of "anti-angiogenesis therapy". An effective means. Cancer cells use a variety of mechanisms to evade strict cell regulation processes, such as cell proliferation, apoptosis, and senescence. Therefore, many tumors can escape the effects of single kinase inhibitors. Sitravatinib enhances tumor immunity by simultaneously inhibiting VEGFR-2 and Axl signaling. Therefore, targeting VEGFRs/Axl is a potential treatment strategy for targeting cancer cells, which can avoid the overcoming of the independent inhibition of VEGFRs and Axl by tumor cells, thus becoming an advanced cancer treatment plan.

Summary of the invention

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

in,

Ring A is selected from 5-membered heteroaryl groups, which contain 2 N heteroatoms;

Alternatively, ring A is selected from 6-membered heteroaryl groups, which contain up to 2 N atoms;

T ₁ and T ₂ are independently selected from C(R ₅ ) and N;

R ₁ and R ₃ are each independently selected from H, F and CH ₃ ;

R ₂ chooses H, F and Cl;

R _{4 is} each independently selected from H, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6 membered heterocycloalkyl, and -LR ₆ , the C _{1-6 6} alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl, 3-6 membered heterocycloalkyl and -LR _{6 are} optionally substituted with 1, 2 or 3 R;

R _{5 is} selected from H and Cl;

R _{6 is} selected from H, OH, 5- to 6-membered heterocycloalkyl and C _1-3 alkyl;

L is selected from -CH ₂ -, -CH ₂ -CH ₂ -and

n is selected from 0, 1, 2 and 3;

R is independently selected from F, Cl, Br, I, OH, CN, COOH, NH ₂ , -NHCH ₃ , -N(CH3) ₂ , CH ₃ , CH ₂ CH ₃ , CF ₃ , -OCH ₃ ,- OCH ₂ CH ₃ , -O-CH(CH ₃ ) ₂ , -C(=O)OCH ₃ , -C(=O)CH ₃ and -C(=O)CH ₂ CH ₃ ;

The 5-membered heteroaryl, 6-membered heteroaryl, C _1-6 heteroalkyl and 3-6 heterocycloalkyl each independently comprise 1, 2 or 3 independently selected from O, S, N and NH heteroatom or heteroatom group.

In some aspects of the present invention, the compound of the above formula (I) or a pharmaceutically acceptable salt thereof, wherein:

T ₁ and T ₂ are independently selected from C(R ₅ ) and N;

R ₁ and R ₃ are each independently selected from H, F and CH ₃ ;

R _{2 is} selected from H, F and Cl; R _{4 is} each independently selected from H, C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl, The C _1-6 alkyl, C _1-6 heteroalkyl, C _3-6 cycloalkyl and 3-6 membered heterocycloalkyl are optionally substituted with 1, 2 or 3 R;

R _{5 is} selected from H and Cl;

n is selected from 0, 1, 2 and 3;

R are independently selected from F, Cl, Br, I, OH, CN, COOH, NH2, -NHCH3, -N(CH3)2, CH3, CH2CH3, CF3, -OCH3, -OCH2CH3, -O-CH(CH3 ) 2, -C(=O)OCH3, -C(=O)CH3 and -C(=O)CH2CH3.

In some embodiments of the present invention, the above compound or a pharmaceutically acceptable salt thereof, wherein ring A is selected from pyridyl, pyrazolyl and imidazolyl, and other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof, wherein R _{4 is} independently selected from H, C _1-4 alkyl, C _1-4 heteroalkyl, 5-6 membered hetero Cycloalkyl and -LR ₆ , said C _1-4 alkyl, C _1-4 heteroalkyl, 5-6 membered heterocycloalkyl and -LR _{6 are} optionally substituted by 1, 2 or 3 R, Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof, wherein R _{4 is} independently selected from H,

Said

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

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof, wherein R _{4 is} independently selected from H, C _1-4 alkyl and C _1-4 heteroalkyl, the C _{1-4 The 4-} alkyl group and C _1-4 heteroalkyl group are optionally substituted with 1, 2 or 3 R, and other variables are as defined in the present invention.

Said

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof, wherein the structural unit

Selected from

Other variables are as defined in the present invention.

Selected from

Other variables are as defined in the present invention.

Selected from

Other variables are as defined in the present invention.

Selected from

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from

in,

R _{41 is} selected from H, C _1-4 alkyl, C _1-4 heteroalkyl, 6-membered heterocycloalkyl and -LR ₆ , the H, C _1-4 alkyl, C _1-4 heteroalkyl , 6-membered heterocycloalkyl and -LR _{6 are} optionally substituted with 1, 2 or 3 R;

R _{42 is} selected from H and -LR ₆ , said -LR _{6 is} optionally substituted by 1, 2 or 3 R;

R _{6 is} selected from H, OH, 6-membered heterocycloalkyl and C _1-3 alkyl;

R ₂ , L and R are as defined in the present invention.

R ₁ , R ₂ , R ₃ and R _{4 are} as defined in the present invention.

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

There are also some technical solutions of the present invention that are arbitrarily combined with the above-mentioned variables.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned compound or a pharmaceutically acceptable salt as an active ingredient and a pharmaceutically acceptable carrier.

The present invention also provides the application of the compound or pharmaceutically acceptable salt or the above composition in the preparation of Axl and VEGFR-2 inhibitors.

In some aspects of the present invention, the above-mentioned Axl and VEGFR-2 inhibitors are drugs for the treatment of cancer.

Technical effect

The compounds of the present invention can be used to treat tumor diseases that respond to the inhibition of protein tyrosine kinase activity, such as cancers that respond to inhibition of the protein tyrosine kinase activity of growth factor receptors. The present invention also provides new compounds and methods for the treatment of diseases that are responsive to the inhibition of receptor-type tyrosine kinase signaling, for example, cancers that are responsive to VEGF receptor signaling inhibition and Axl kinase inhibition. . The compound of the present invention has excellent in vitro activity and has good inhibitory activity on VEGFR2/KDR kinase and AXL kinase. Compared with the prior art, the representative compound of the present invention has greatly improved permeability, more stable performance in metabolic stability, and excellent performance in CYP inhibitory properties. The compound of the present invention has good PK properties in vivo, and has good exposure and bioavailability. The compound of the present invention has better drug-making properties and better curative effect in clinical practice.

Definition and description

Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A specific term or phrase should not be considered uncertain or unclear without a special definition, but should be understood in its ordinary meaning. When a trade name appears in this article, it is meant to refer to its corresponding commodity or its active ingredient.

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

The term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention, which is prepared from a compound with specific substituents discovered in the present invention and a relatively non-toxic acid or base. When the compound of the present invention contains a relatively acidic functional group, a base addition salt can be obtained by contacting the compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salt or similar salts. When the compound of the present invention contains a relatively basic functional group, the acid addition salt can be obtained by contacting the compound with a sufficient amount of acid in a pure solution or 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, hydrogen carbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, the organic acid includes, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid; 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 basic and acidic functional groups, which can be converted into any base or acid addition salt.

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

The term "optional" or "optionally" refers to the event or condition described later that may but not necessarily occur, and the description includes a situation in which the event or condition occurs and a situation in which the event or condition does not occur .

The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by substituents, and may include deuterium and hydrogen variants, as long as the valence of the specific atom is normal and the substituted compound is stable of. When the substituent is oxygen (ie =O), it means that two hydrogen atoms are replaced.

The term "optionally substituted" means that it can be substituted or unsubstituted. Unless otherwise specified, the type and number of substituents can be arbitrary on the basis that they can be chemically realized.

When any variable (such as 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 to 2 Rs, the group can be optionally substituted with up to two Rs, and R has independent options in each case. In addition, combinations of substituents and/or variants thereof are only permitted 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, and -C ₀ alkyl-A means that the structure is actually -A.

When the number of a substituent is 0, it means that the substituent is absent. For example, -A-(R) ₀ means that the structure is actually -A.

When a substituent is vacant, it means that the substituent is absent. For example, when X in A-X is vacant, it means that the structure is actually A.

When one of the variables is selected from a single bond, it means that the two connected groups 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 the listed substituents do not indicate which atom is connected to the substituted group, such substituents can be bonded via any atom. For example, a pyridyl group can pass through any one of the pyridine ring as a substituent. The carbon atom is attached to the substituted group.

When the bond of a substituent can be cross-connected to two or more atoms on a ring, the substituent can be bonded with any atom on the ring, for example, a structural unit

It means that the substituent R can be substituted at any position on the cyclohexyl or cyclohexadiene.

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

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

It can also be formed by connecting 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 only permitted if such combinations result in stable compounds.

Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the connection method of the chemical bond is not positioned, and there is a H atom at the connectable site, when the chemical bond is connected, the number of H atoms at the site will correspondingly decrease with the number of chemical bonds connected to become the corresponding valence number的组。 The group. The chemical bond between the site and other groups can be a straight solid bond

Straight dashed key

Or wavy line

Express. For example _{, the straight solid bond in -OCH 3} means that it is connected to other groups through the oxygen atom in the group;

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

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

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

These four connection methods, even though the H atom is drawn on -N-, but

Still include

The group in this connection mode, only when one chemical bond is connected, the H at this position will decrease by one and become the corresponding monovalent piperidinyl group.

Unless otherwise specified, the number of atoms in a ring is usually defined as the number of members of the ring. For example, a "5- to 7-membered ring" refers to a "ring" in which 5 to 7 atoms are arranged around.

Unless otherwise specified, the term "C _1-6 alkyl" is used to indicate a linear or branched saturated hydrocarbon group composed of 1 to 6 carbon atoms. The C _1-6 alkyl group includes C _1-5 , C _1-4 , C _1-3 , C _1-2 , C _2-6 , C _2-4 , C ₆ and C ₅ alkyl, etc.; it may Is monovalent (such as methyl), divalent (such as methylene) or multivalent (such as methine). Examples of C _1-6 alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl) , S-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc.

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

The term "heteroalkyl" by itself or in combination with another term means a stable linear or branched alkyl group or a combination thereof composed of a certain number of carbon atoms and at least one heteroatom or heteroatom group. In some embodiments, the heteroatoms are selected from B, O, N, and S, wherein nitrogen and sulfur atoms are optionally oxidized, and nitrogen heteroatoms are optionally quaternized. In other embodiments, the heteroatom group is selected from -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O), -S(=O) ₂ -, -C(=O)N(H)-, -N(H)-, -C(=NH)-, -S(=O) ₂ N(H)- and -S(=O)N( H)-. In some embodiments, the heteroalkyl group is a C _1-6 heteroalkyl group; in other embodiments, the heteroalkyl group is a C _1-4 heteroalkyl group. The heteroatom or heteroatom group can be located in any internal position of the heteroalkyl group, including the connection position of the alkyl group to the rest of the molecule, but the terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkane The oxy) is a customary expression and refers to those alkyl groups that are connected to the rest of the molecule through an oxygen atom, an amino group, or a sulfur atom, respectively. Examples of heteroalkyl groups include, but are not limited to, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ CH ₃ , -OCH ₂ (CH ₃ ) ₂ , -CH ₂ -CH ₂ -O-CH ₃ , -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )(CH ₂ CH ₃ ), -CH ₂ -CH ₂ -NH- CH ₃ , -CH ₂ -CH ₂ -N( CH ₃ ) -CH ₃ , -SCH ₃ , -SCH ₂ CH ₃ , -SCH ₂ CH ₂ CH ₃ , -SCH ₂ (CH ₃ ) ₂ , -CH ₂ -S-CH ₂ -CH ₃ , -CH _2- CH ₂ , -S(=O)-CH ₃ , -CH ₂ -CH ₂ -S(=O) ₂ -CH ₃ . Up to two heteroatoms can be continuous, for example -CH ₂ -NH-OCH ₃ .

Unless otherwise specified, "C _3-6 cycloalkyl" means a saturated cyclic hydrocarbon group composed of 3 to 6 carbon atoms, which is a monocyclic and bicyclic ring system. The C _3-6 cycloalkyl includes C _{3 to 5} , C _{4 to 5} and C _{5 to 6} cycloalkyl, etc.; it can be monovalent, divalent or multivalent. Examples of C _3-6 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

Unless otherwise specified, the term "3- to 6-membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 3 to 6 ring atoms, with 1, 2, 3 or 4 ring atoms. Are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein nitrogen atoms are optionally quaternized, and carbon, nitrogen and sulfur heteroatoms may optionally be oxidized (ie C(=O), NO and S(O)p, p is 1 or 2). It includes monocyclic and bicyclic ring systems, where the bicyclic ring system includes spiro, fused, and bridged rings. In addition, with regard to the "3- to 6-membered heterocycloalkyl group", a heteroatom may occupy the connection position between the heterocycloalkyl group and the rest of the molecule. The 3- to 6-membered heterocycloalkyl group includes 4- to 6-membered, 5- to 6-membered, 4-, 5-, and 6-membered heterocycloalkyl. Examples of 3- to 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-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), Dioxanyl, dithiazinyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, or homopiperazinyl Pyridyl and so on.

Unless otherwise specified, the term "5- to 6-membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group consisting of 5 to 6 ring atoms, with 1, 2, 3 or 4 ring atoms. Are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein nitrogen atoms are optionally quaternized, and carbon, nitrogen and sulfur heteroatoms can be optionally oxidized (ie C(=O), NO And S(O)p, p is 1 or 2). It includes monocyclic and bicyclic ring systems, where the bicyclic ring system includes spiro, fused, and bridged rings. In addition, with regard to the "5- to 6-membered heterocycloalkyl group", a heteroatom may occupy the connection position between the heterocycloalkyl group and the rest of the molecule. The 5- to 6-membered heterocycloalkyl group includes 5- and 6-membered heterocycloalkyl groups. Examples of 5- to 6-membered heterocycloalkyl groups include, but are not limited to, 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-piperidinyl, etc.), piperazinyl (including 1 -Piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithiaalkyl, isoxazolidinyl, isothiazole Alkyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl, homopiperazinyl or homopiperidinyl, etc.

Unless otherwise specified, the term "6-membered heterocycloalkyl" by itself or in combination with other terms means a saturated cyclic group composed of 6 ring atoms, of which 1, 2, 3 or 4 ring atoms are independently selected from O, S and N heteroatoms, the rest are carbon atoms, wherein nitrogen atoms are optionally quaternized, carbon, nitrogen and sulfur heteroatoms can optionally be oxidized (ie C(=O), NO and S(O) )p, p is 1 or 2). It includes monocyclic and bicyclic ring systems, where the bicyclic ring system includes spiro, fused, and bridged rings. In addition, with regard to the "6-membered heterocycloalkyl group", a heteroatom may occupy the position of attachment of the heterocycloalkyl group to the rest of the molecule. Examples of 6-membered heterocycloalkyl include, but are not limited to, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperidinyl). Piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazine Base and so on.

Unless otherwise specified, the terms "5-6 membered heteroaryl ring" and "5-6 membered heteroaryl group" can be used interchangeably in the present invention. The term "5-6 membered heteroaryl group" means a ring consisting of 5 to 6 ring atoms. It is composed of a monocyclic group with a conjugated π-electron system, in which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atom is optionally For quaternization, carbon, nitrogen, and sulfur heteroatoms can optionally be oxidized (ie, C(=O), NO, and S(O)p, where p is 1 or 2). The 5- to 6-membered heteroaryl group can be connected to the rest of the molecule through a heteroatom or a carbon atom. The 5- to 6-membered heteroaryl group includes 5-membered and 6-membered heteroaryl groups. Examples of the 5- to 6-membered heteroaryl group include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-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, etc.) , 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, the terms "6-membered heteroaromatic ring" and "6-membered heteroaryl" can be used interchangeably in the present invention. The term "6-membered heteroaryl" refers to a conjugated π-electron system composed of 6 ring atoms. A monocyclic group of which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atoms are optionally quaternized, carbon, nitrogen and The sulfur heteroatom may optionally be oxidized (ie, C(=O), NO, and S(O)p, where p is 1 or 2). The 6-membered heteroaryl group can be attached to the rest of the molecule through a heteroatom or a carbon atom. Examples of the 6-membered heteroaryl include, but are not limited to, pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidine) Base etc.).

Unless otherwise specified, the terms "5-membered heteroaromatic ring" and "5-membered heteroaryl" can be used interchangeably in the present invention. The term "5-membered heteroaryl" refers to a conjugated π-electron system composed of 5 ring atoms. A monocyclic group of which 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the nitrogen atoms are optionally quaternized, carbon, nitrogen and The sulfur heteroatom may optionally be oxidized (ie, C(=O), NO, and S(O)p, where p is 1 or 2). The 5-membered heteroaryl group can be attached to the rest of the molecule through a heteroatom or a carbon atom. Examples of the 5-membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.) Etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazole) Base, 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.).

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 listed below, the embodiments formed by combining them with other chemical synthesis methods, and those well known to those skilled in the art Equivalent alternatives, preferred implementations include but are not limited to the embodiments of the present invention.

The solvent used in the present invention is commercially available.

The present invention uses the following abbreviations: FA stands for formic acid; TFA stands for trifluoroacetic acid; ACN stands for acetonitrile; DIEA stands for diisopropylethylamine; P(o-tolyl) ₃ stands for tris(o-methylphenyl)phosphine ; TBAI stands for tetrabutylammonium iodide; T3P stands for 1-propyl phosphoric anhydride; TBSCl stands for tert-butyldimethylchlorosilane; HATU stands for 2-(7-azabenzotriazole)-N,N, N',N'-tetramethylurea hexafluorophosphate; HOBt stands for 1-hydroxybenzotriazole; PyBOP stands for hexafluorophosphate benzotriazol-1-yl-oxytripyrrolidinyl phosphorus; DMSO stands for two Methyl sulfoxide; ATP stands for adenosine triphosphate.

Compounds are based on conventional naming principles in the field or use

The software is named, and the commercially available compounds use the supplier catalog name.

Detailed ways

The present invention will be described in detail through the following examples, but it is not meant to impose any disadvantageous restriction on the present invention. The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and those well known to those skilled in the art Equivalent alternatives, preferred implementations include but are not limited to the embodiments of the present invention. It will be obvious to those skilled in the art that various changes and improvements can be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

Example 1: Compound 1

Step A: Compound 1-1 (12g, 91.23mmol), 1-2 (14.05g, 91.23mmol), tetrakis (triphenylphosphorus) palladium (5.27g, 4.56mmol), sodium carbonate (19.34g, 182.46mmol) ) Was dissolved in 1,4-dioxane (100 mL) and water (20 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 110°C for 12 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate=20/1～5/1) to obtain 1-3. MS(ESI) m/z: 124.0 [M+H ⁺ ].

Step B: Combine compound 1-3 (1g, 8.12mmol), 1-4 (2.8g, 8.12mmol), palladium acetate (0.18g, 0.812mmol), cesium carbonate (5.29g, 16.24mmol), tetrabutyl bromide The amine (2.62 g, 8.12 mmol) was dissolved in N,N-dimethylacetamide (30 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 130°C for 12 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product is separated by high performance liquid chromatography (

40g

Fast silica gel column, the eluent is 0-50% ACN/H ₂ O, the flow rate is 60 mL/min) to obtain 1-5. MS(ESI) m/z: 387.9 [M+H ⁺ ].

Step C: Dissolve compound 1-5 (0.2g, 0.516mmol), 1-6 (0.078g, 0.619mmol), potassium carbonate (0.142g, 1.03mmol) in N-methylpyrrolidone (5mL). Stir in microwave at 150°C for 2 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phases were washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product is separated by high performance liquid chromatography (

40g

Fast silica gel column, the eluent is 0-50% ACN/H ₂ O, the flow rate is 60 mL/min) to obtain 1-7. MS(ESI) m/z: 495.0 [M+H ⁺ ].

Step D: Compound 1-8 (0.3g, 1.34mmol) was dissolved in thionyl chloride (5mL), the resulting mixture was stirred at 25°C for 12 hours. Concentrated under reduced pressure to obtain the crude product, and then combined with 1-7 (25mg, 50.55 μmol) was dissolved in dichloromethane (5 mL), DIEA (26.13 mg, 202.20 μmol) was added to the solution, and the resulting mixture was stirred at 25° C. for 1 hour. The reaction solution was quenched by adding water (50 mL) and extracted with dichloromethane (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 1-9. MS (ESI) m/z: 700.2 [M+H ⁺ ].

Step E: Dissolve compound 1-9 (45 mg, 64.31 μmol) in dichloromethane (1 mL), then add trifluoroacetic acid (770 mg, 6.75 mmol, 0.5 mL) to the reaction solution, and stir the resulting mixture at 25°C 0.5 hours. The reaction solution was quenched by adding sodium bicarbonate aqueous solution (30 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phases were washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and then depressurized Concentrate to obtain crude product. The crude product was purified by (column information: Phenomenex Synergi C18 150*25*10 μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 17%-41%, 8 minutes) to obtain compound 1. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.38(s,1H),10.00(s,1H),8.53(d,J=2.0Hz,1H), 8.48(d,J=5.6Hz,1H) ,7.88(dd,J＝13.2,2.4Hz,1H),7.75(dd,J＝8.0,2.0Hz,1H),7.72-7.57(m,5H),7.50(dd,J＝8.8,1.2Hz,1H) ), 7.39-7.33 (m, 1H), 7.19 (d, J = 2.4 Hz, 1H), 7.18-7.13 (m, 2H), 6.80 (dd, J = 5.6, 2.4 Hz, 1H), 3.76 (s, 2H)3.41(br s,2H),3.24(s,3H),2.69-2.64(m,2H),1.47(br d,J=10.0Hz,4H).MS(ESI)m/z:600.1[M +H ⁺ ].

Example 2: Compound 2

Step A: Dissolve compound 2-1 (2g, 13.42mmol), 2-2 (2.11g, 13.42mmol), potassium carbonate (3.71g, 26.85mmol) in N,N-dimethylformamide (20mL) The resulting mixture was stirred at 80°C for 3 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 2-3. MS(ESI)m/z:269.7[M+H ⁺ ].

Step B: Compound 2-3 (1.9g, 7.05mmol), 1-2 (1.19g, 7.75mmol), tetrakis(triphenylphosphorus) palladium (814.33mg, 704.71μmol), sodium carbonate (1.49g, 14.09 mmol) was dissolved in 1,4-dioxane (20 mL) and water (4 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 80°C for 3 hours. Water (100 mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (100 mL×2). The combined organic phases were washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 20/1 to 5/1) to obtain 2-4. MS(ESI)m/z:261.8[M+H ⁺ ].

Step C: Compound 2-4 (0.5g, 1.91mmol), 1-4 (0.66g, 1.91mmol), palladium acetate (42.97mg, 0.191mmol), triethylamine (387.39mg, 3.83mmol), tetrabutyl Amine bromide (617.07 mg, 1.91 mmol), P(o-tolyl) ₃ (582.61 mg, 1.91 mmol) were dissolved in toluene (10 mL), the resulting mixture was replaced with nitrogen three times and stirred at 90°C for 12 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate=20/1～5/1) to obtain 2-5. MS(ESI) m/z: 526.1 [M+H ⁺ ].

Step D: Dissolve compound 2-5 (90 mg, 171.26 mmol) in ethanol (10 mL), then add iron powder (191.28 mg, 3.43 mmol), ammonium chloride (18.32 mg, 0.342 mmol), and the resulting mixture at 100°C Stir for 12 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain a crude product, which was extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 2-6. MS(ESI) m/z: 496.1 [M+H ⁺ ].

Step E: Compound 1-8 (0.3 g, 1.34 mmol) was dissolved in thionyl chloride (5 mL), and the resulting mixture was stirred at 25°C for 12 hours. Concentrate under reduced pressure to obtain crude acid chloride. 2-6 (80mg, 161.44μmol) and crude acid chloride (78mg, 322.88μmol) were dissolved in tetrahydrofuran (5mL). DIEA (83.46mg, 645.75μmol) was added to the solution. Stir at °C for 1 hour. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by a reverse phase column (

40g

Fast silica gel column, the eluent is 0-50% ACN/0.1% FA-H ₂ O, the flow rate is 60 mL/min) to obtain 2-7. MS(ESI) m/z: 701.2 [M+H ⁺ ].

Step F: Compound 2-7 (72 mg, 102.75 μmol) was dissolved in dichloromethane (1 mL), and then trifluoroacetic acid (1.23 g, 10.79 mmol, 0.8 mL) was added to the reaction solution, and the resulting mixture was kept at 25°C Stir for 0.5 hour. To the reaction solution was added an aqueous sodium hydrogen carbonate solution (30 mL) to quench the reaction and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (column information: Phenomenex Synergi C18 150*25*10μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 20%-40%, 10 minutes) to obtain compound 2 Of formate. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.34(s,1H), 10.03(s,1H), 8.72(s,1H), 8.61(d,J=2.0Hz,1H), 8.19(s, 1H),7.95(d,J=15.6Hz,1H),7.85-7.78(m,2H),7.72-7.63(m,4H),7.46-7.44(m,2H),7.37-7.32(m,1H) ,7.16(t,J＝8.88Hz,2H), 3.82(s,2H), 3.43(t,J＝8.8Hz,2H), 3.25(s,3H), 2.71(t,J＝5.6Hz,2H) ,1.47(br d,J=2.8Hz,4H); MS(ESI)m/z:601.1[M+H ⁺ ]

Example 3: Compound 3

Step A: Dissolve compound 3-1 (3.4g, 23.13mmol), 3-2 (3.18g, 25.45mmol), cesium carbonate (11.31g, 34.70mmol), TBAI (1.71g, 4.63mmol) in N, N -In dimethylacetamide (15 mL), the resulting mixture was stirred at 90°C for 12 hours. Water (50mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (50mL×2). The combined organic phases were washed with saturated sodium chloride solution (50mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude compound 3-3. MS(ESI)m/z:191.1[M+H ⁺ ].

Step B: Dissolve compound 3-3 (4.4g, 23.03mmol), TBSCl (4.17g, 27.64mmol), imidazole (3.14g, 46.07mmol) in N,N-dimethylacetamide (15mL) to obtain The mixture was stirred at 25°C for 12 hours. Water (50mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (50mL×2). The combined organic phases were washed with saturated sodium chloride solution (50mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by reverse phase column (waters Xbridge BEH C18 250*50mm*10μm; mobile phase: [water (0.05% ammonia v/v)-ACN]; ACN%: 65%-80%, 20 minutes) to obtain 3- 4. MS(ESI)m/z:304.9[M+H ⁺ ].

Step C: Dissolve compound 1-1 (10g, 76.03mmol), 2-2 (10g, 76.03mmol), potassium carbonate (21.01g, 152.05mmol) in N,N-dimethylacetamide (20mL), The resulting mixture was stirred at 100°C for 3 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 20/1, 5/1) to obtain compound 3-5. MS(ESI)m/z:268.7[M+H ⁺ ].

Step D: The compound 3-5 (5g, 18.61mmol), 1-2 (3.44g, 22.34mmol), tetrakis (triphenylphosphorus) palladium (2.15g, 1.86mmol), sodium carbonate (3.95g, 37.23mmol) ) Was dissolved in dioxane (50 mL) and water (10 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 80°C for 3 hours. Water (100 mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (100 mL×2). The combined organic phases were washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 20/1, 5/1) to obtain 3-6. MS(ESI)m/z:260.8[M+H ⁺ ].

Step E: Compound 3-6 (255.71mg, 982.69mmol), 3-4 (300mg, 982.69mmol), palladium acetate (22.06mg, 98.27μmol,), triethylamine (497.19mg, 4.91mmol), lithium bromide ( 256.02 mg, 2.95 mmol), P(o-tolyl) ₃ (299.10 mg, 982.69 mmol) was dissolved in toluene (10 mL), the resulting mixture was replaced with nitrogen three times and stirred at 110°C for 12 hours. Water (100mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (100mL×2). The combined organic phases were washed with saturated sodium chloride solution (25mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain Crude. The crude product passes through the reverse phase column. Column information: waters Xbridge BEH C18 250*50mm*10μm; mobile phase: [water (0.05% ammonia v/v)-ACN]; ACN%: 55%-85%, purified in 20 minutes to obtain 3 -7. MS(ESI)m/z:485.0[M+H ⁺ ].

Step F: Dissolve compound 3-7 (200mg, 412.72μmol) in ethanol (10mL) and water (2mL), then add iron powder (230.48mg, 4.13mmol) and ammonium chloride (110.38mg, 2.06mmol) to obtain The mixture was stirred at 90°C for 6 hours. The reaction solution was filtered and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 3-8. MS(ESI)m/z:455.0[M+H ⁺ ].

Step G: Dissolve compound 3-8 (90mg, 197.97μmol) in dichloromethane (5mL), and add 1-8 (88.3mg, 395.94μmol), DIEA (127.93mg, 989.86 μmol) and T3P (377.94 mg, 593.91 μmol). The resulting mixture was stirred at 25°C for 12 hours. Water (50 mL) was added to the reaction solution and extracted with ethyl acetate (50 mL×2). The combined organic phases were washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product 3-9. MS(ESI)m/z:660.2[M+H ⁺ ].

Step H: Compound 3-9 (65mg, 98.52μmol) was dissolved in dichloromethane (2mL), then trifluoroacetic acid (1.54g, 13.51mmol) was added to the reaction solution, and the resulting mixture was stirred at 25°C for 0.5 hours . The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by (Phenomenex Synergi C18 150*25*10 μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 17%-50%, 10 minutes) to obtain the formate salt of compound 3. 1H NMR(400MHz,DMSO-d6)δ=10.39(s,1H),10.01(s,1H),8.40(d,J=5.6Hz,1H),8.22(s,1H),7.97(s,1H) ,7.87(dd,J=2.4,13.2Hz,1H),7.77(s,1H),7.70-7.60(m,2H),7.54(s,1H),7.50-7.46(m,1H),7.39-7.31 (m,1H),7.21-7.11(m,2H),6.92(d,J=16.0Hz,1H), 6.86(d,J=2.4Hz,1H), 6.74(dd,J=2.4,5.6Hz, 1H), 4.12 (t, J = 5.6 Hz, 2H), 3.72 (t, J = 5.6 Hz, 2H), 1.47 (br d, J = 7.2 Hz, 4H); MS (ESI) m/z: 546.1[ M+H ⁺ ].

Example 4: Compound 4

Step A: Dissolve compound 3-1 (1g, 6.80mmol), 4-1 (1.46g, 10.21mmol), cesium carbonate (3.33g, 10.21mmol) in N,N-dimethylformamide (8mL) The resulting mixture was stirred at 60°C for 1 hour. Water (20mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (30mL×3). The combined organic phases were washed with saturated sodium chloride solution (50mL×3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain Compound 4-2, the crude product was used directly in the next step.

Step B: Compound 4-2 (1.43 g, 6.83 mmol) and compound 4-3 (1.54 g, 20.48 mmol) were dissolved in DMSO (5 mL), and the resulting mixture was stirred at 60° C. for 5 hours. The reaction solution was diluted with 1mol/L hydrochloric acid (10mL) and ethyl acetate (20mL), separated, the aqueous phase was washed with ethyl acetate (30mL), the organic phase was discarded, and the aqueous phase was adjusted with 1mol/L sodium hydroxide to adjust the pH= 8 and extracted with ethyl acetate (30 mL×3)), the combined organic phases were washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 4-4. MS(ESI)m/z:247.8[M+H ⁺ ].

Step C: Compound 4-4 (900 mg, 3.63 mmol) was dissolved in tetrahydrofuran (10 mL), di-tert-butyl dicarbonate (949.97 mg, 4.35 mmol,) was added, and the resulting mixture was stirred at 20°C for 12 hours. Concentrate under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (ethyl acetate/petroleum ether=0%-50%) to obtain compound 4-5.

Step E: Compound 3-6 (0.4g, 1.54mmol), 4-5 (1.07g, 3.07mmol), palladium acetate (69.02mg, 307.43μmol), triethylamine (933.27mg, 9.22mmol), lithium bromide ( 534.01mg, 6.15mmol), P(o-tolyl) ₃ (467.87mg, 1.54mmol) was dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and stirred at 125°C for 12 Hour. Water (40mL) and ethyl acetate (40mL) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (40mL×3). The combined organic phases were washed with saturated sodium chloride solution (50mL×3), anhydrous It was dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column (ethyl acetate/petroleum ether = 20% to 80%) to obtain 4-6. MS(ESI)m/z:528.3[M+H ⁺ ].

Step F: Dissolve compound 4-6 (150 mg, 284.34 μmol) in ethanol (3 mL), then add iron powder (158.79 mg, 2.84 mmol), saturated ammonium chloride solution (0.3 mL), and stir the resulting mixture at 80°C 2 hours. After filtration, the filtrate was collected and concentrated to remove the organic solvent, then diluted with ethyl acetate (20 mL) and water (20 mL), separated, and the aqueous phase was extracted with ethyl acetate (30 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 4-7, and the crude product was directly used in the next step. MS(ESI)m/z:498.3[M+H ⁺ ].

Step G: Compound 1-8 (72.81mg, 326.20μmol) was dissolved in dichloromethane (5mL), and oxalyl chloride (159.25mg, 1.25mmol) and N,N-dimethylformamide ( 1.83mg, 25.09μmol), reacted at 25°C for half an hour. The solution was concentrated to obtain the corresponding acid chloride and dissolved in dichloromethane (5mL). 4-7 (124.85mg, 250.92μmol) and pyridine (99.24mg, 1.25mmol) were added to the solution at 0°C. The resulting mixture was kept at 25°C. Stir for half an hour. Water (20 mL) was added to the reaction solution for liquid separation, and the aqueous phase was extracted with dichloromethane (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 4-8, and the crude product was directly used in the next step. MS(ESI)m/z:703.2[M+H ⁺ ].

Step H: Compound 4-8 (150 mg, 213.45 μmol) was dissolved in dichloromethane (5 mL), then trifluoroacetic acid (1 mL) was added to the reaction solution, and the resulting mixture was stirred at 20° C. for 1 hour. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative high performance liquid chromatography (column: Waters Xbridge 150*25mm*5μm; mobile phase: [water (0.05% ammonia v/v)-ACN]; B%: 38%-68%, 10 minutes) Compound 4 was obtained. ¹ H NMR (400MHz, DMSO-d6) δ = 10.39 (s, 1H), 10.01 (s, 1H), 8.40 (d, J = 5.6 Hz, 1H), 7.99 (s, 1H), 7.88 (dd, J ＝2.4,13.2Hz,1H),7.77(s,1H),7.65(dd,J＝5.2,9.1Hz,2H),7.54-7.47(m,2H),7.35(t,J＝9.2Hz,1H) ,7.16(t,J=8.8Hz,2H),6.92(d,J=16.0Hz,1H), 6.86(d,J=2.4Hz,1H), 6.74(dd,J=2.4,5.6Hz,1H) , 4.14 (t, J = 6.0 Hz, 2H), 3.35 (t, J = 5.6 Hz, 2H), 3.22 (s, 3H), 2.92 (t, J = 6.0 Hz, 2H), 2.65 (t, J = 5.6 Hz, 2H), 1.54-1.41 (m, 4H); MS (ESI) m/z: 603.2 [M+H ⁺ ].

Example 5: Compound 5

Step A: Dissolve compound 5-1 (1.6g, 8.25mmol) in acetonitrile (20mL), add potassium carbonate (2.28g, 16.50mmol), potassium iodide (136.93mg, 824.86μmol) and 5-2 (1.94g, 8.66 mmol), and the resulting mixture was stirred at 80°C for 12 hours. Water (100 mL) was added to the reaction solution, a white solid precipitated out, filtered and collected the filter cake, and concentrated under reduced pressure to obtain compound 5-3. MS(ESI)m/z:338.2[M+H ⁺ ].

Step B: Combine compound 3-6 (0.4g, 1.54mmol), compound 5-3 (1.04g, 3.08mmol), palladium acetate (69.15mg, 308.00μmol), triethylamine (935.00mg, 9.24mmol), lithium bromide (401.25mg, 4.62mmol), P(o-tolyl) ₃ (468.73mg, 1.54mmol) was dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and stirred at 125°C 36 hours. Water (40mL) and ethyl acetate (40mL) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (40mL×3). The combined organic phases were washed with saturated sodium chloride solution (50mL×3), anhydrous It was dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column (ethyl acetate/petroleum ether=20%-80%) to obtain 5-4. MS(ESI)m/z:470.0[M+H ⁺ ].

Step C: Dissolve compound 5-4 (370 mg, 788.13 μmol) in ethanol (6 mL), then add iron powder (440.17 mg, 7.88 mmol), saturated ammonium chloride solution (0.5 mL), and stir the resulting mixture at 80°C 2 hours. After filtration, the filtrate was collected and concentrated to remove the organic solvent, then diluted with ethyl acetate (20 mL) and water (20 mL), separated, and the aqueous phase was extracted with ethyl acetate (30 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 5-5, and the crude product was directly used in the next step. MS(ESI)m/z:440.3[M+H ⁺ ].

Step D: Dissolve compound 1-8 (211.27mg, 946.57μmol) in dichloromethane (10mL), add oxalyl chloride (462.11mg, 3.64mmol, 318.69μL) and N,N-dimethyl at 0°C Formamide (5.32mg, 72.81μmol), reacted at 25°C for half an hour. The solution was concentrated to obtain the corresponding acid chloride and dissolved in dichloromethane (5mL), 5-5 (320mg, 728.13μmol) and pyridine (287.98mg, 3.64mmol) were added to the solution at 0℃. The resulting mixture was stirred at 25℃ half an hour. Water (20 mL) was added to the reaction solution for liquid separation, and the aqueous phase was extracted with dichloromethane (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 5-6, and the crude product was directly used in the next step. MS(ESI)m/z:645.2[M+H ⁺ ].

Step E: Compound 5-6 (450 mg, 698.04 μmol) was dissolved in dichloromethane (20 mL), then trifluoroacetic acid (2 mL) was added to the reaction solution, and the resulting mixture was stirred at 20° C. for 3 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative high performance liquid chromatography (column: Phenomenex Synergi C18 150*25*10μm; mobile phase: [water (0.225% FA)-ACN]; B%: 8%-38%, 10 minutes) to obtain the compound 5 formate. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.40 (br d, J = 2.4 Hz, 1H), 10.01 (br s, 1H), 8.40 (d, J = 5.6 Hz, 1H), 8.30 (br s ,1H),8.02(s,1H),7.91-7.81(m,2H),7.65(dd,J=5.0,9.2Hz,2H),7.58-7.44(m,2H),7.35(t,J=9.2 Hz, 1H), 7.16 (t, J = 8.8 Hz, 2H), 6.95 (d, J = 16.0 Hz, 1H), 6.87 (d, J = 2.4 Hz, 1H), 6.75 (dd, J = 2.4, 5.6 Hz,1H),4.22(br t,J=6.0Hz,2H),3.16-3.03(m,2H),1.55-1.38(m,4H); MS(ESI)m/z:545.3[M+H ⁺ ].

Example 6: Compound 6

Step A: Dissolve compound 3-1 (7g, 47.63mmol) in tetrahydrofuran (100mL), add sodium hydride (2.48g, 61.92mmol, 60% content) in portions at 0°C and stir at 0°C for 30 minutes, 6-1 (9.93 g, 59.53 mmol) was added, and the resulting mixture was stirred at 25°C for 12 hours. Saturated ammonium chloride (50mL) was carefully added to quench the reaction at 0°C, the layers were separated, the aqueous phase was extracted with ethyl acetate (50mL×3), the organic phases were combined, washed with saturated sodium chloride (50mL), and anhydrous sulfuric acid It was dried over sodium, filtered, and concentrated under reduced pressure to obtain compound 6-2.

Step B: Combine compound 6-2 (1.28 g, 4.61 mmol), compound 3-6 (1 g, 3.84 mmol), palladium acetate (86.28 mg, 384.29 μmol), triethylamine (1.94 g, 19.21 mmol), lithium bromide ( 1.00g, 11.53mmol), P(o-tolyl) ₃ (1.17g, 3.84mmol) was dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and stirred at 125°C 36 Hour. Water (50mL) and ethyl acetate (50mL) were added to the reaction solution, and the layers were separated. The aqueous phase was extracted with ethyl acetate (50mL×3). The combined organic phases were washed with saturated sodium chloride solution (50mL) and anhydrous sodium sulfate Dry, filter and concentrate under reduced pressure to obtain the crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=10/1～2/1) to obtain 6-3. MS(ESI)m/z:457.2[M+H ⁺ ].

Step C: Compound 6-3 (580mg, 1.27mmol) was dissolved in ethanol (7mL), then iron powder (709.47mg, 12.70mmol) and saturated ammonium chloride solution (0.7mL) were added, and the resulting mixture was stirred at 80°C 10 hours. After filtration, the filtrate was collected and concentrated to remove the organic solvent, then diluted with ethyl acetate (30 mL) and water (30 mL), separated, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 6-4, and the crude product was directly used in the next step. MS(ESI)m/z:427.1[M+H ⁺ ].

Step D: Dissolve compound 1-8 (285.70mg, 1.28mmol) in dichloromethane (5mL), add oxalyl chloride (624.87mg, 4.92mmol) and N,N-dimethylformamide ( 7.20mg, 98.46μmol), react at 25°C for half an hour. The solution was concentrated to obtain the corresponding acid chloride and dissolved in dichloromethane (5mL), 6-4 (420mg, 984.63μmol) and pyridine (389.42mg, 4.92mmol) were added to the solution at 0℃. The resulting mixture was stirred at 25℃ half an hour. Water (30 mL) and dichloromethane (30 mL) were added to the reaction solution for liquid separation, and the aqueous phase was extracted with dichloromethane (50 mL×3). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 6-5, and the crude product was directly used in the next step. MS(ESI)m/z:632.3[M+H ⁺ ].

Step E: Compound 6-5 (700 mg, 1.11 mmol) was dissolved in dichloromethane (14 mL), then trifluoroacetic acid (7 mL) was added to the reaction solution, and the resulting mixture was stirred at 25° C. for 2 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product passed preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 150*25mm*5μm; mobile phase: [water (0.05% ammonia v/v)-ACN]; B%: 36%-66%, 10 minutes ) Purification to obtain compound 6. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 12.94 (br s, 1H), 10.38 (s, 1H), 10.00 (s, 1H), 8.40 (d, J = 5.6 Hz, 1H), 8.09-7.76 (m, 3H), 7.65 (dd, J = 5.2, 9.2 Hz, 2H), 7.56 (d, J = 16.0 Hz, 1H), 7.49 (dd, J = 1.2, 8.8 Hz, 1H), 7.35 (t, J = 9.2 Hz, 1H), 7.16 (t, J = 8.8 Hz, 2H), 6.94 (d, J = 16.0 Hz, 1H), 6.84 (d, J = 2.4 Hz, 1H), 6.75 (dd, J = 2.4, 5.6 Hz, 1H), 1.53-1.43 (m, 4H); MS (ESI) m/z: 502.3 [M+H ⁺ ].

Example 7: Compound 7

Step A: Dissolve compound 1-1 (3.5g, 26.61mmol), 7-1 (3.70g, 26.61mmol), potassium carbonate (7.36g, 53.22mmol) in N,N-dimethylformamide (10mL) The resulting mixture was stirred at 100°C for 12 hours. Water (100 mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (100 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column (petroleum ether: ethyl acetate = 20/1 to 5/1) and purified to obtain 7-2. MS(ESI)m/z:250.8[M+H ⁺ ].

Step B: The compound 7-2 (4.5g, 17.95mmol), 1-2 (3.32g, 21.55mmo), tetrakis (triphenylphosphorus) palladium (2.07g, 1.80mmol), sodium carbonate (3.81g, 35.91 mmol) was dissolved in 1,4-dioxane (50 mL) and water (10 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 80°C for 3 hours. The reaction solution was quenched by adding water (50mL) and extracted with ethyl acetate (50mL×2). The combined organic phases were washed with saturated sodium chloride solution (25mL×2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain Crude. The crude product was separated by silica gel column (petroleum ether: ethyl acetate = 1/0 ~ 1/2) and purified to obtain 7-3. MS(ESI)m/z:243.0[M+H ⁺ ].

Step C: The compound 7-3 (1g, 4.13mmol), 3-4 (1.26g, 4.13mmol), palladium acetate (92.68mg, 412.83μmol), triethylamine (2.09g, 20.64mmol), lithium bromide (1.08 g, 12.38 mmol) was dissolved in toluene (15 mL), the resulting mixture was replaced with nitrogen three times and stirred at 120°C for 12 hours. Water (100 mL) was added to the reaction solution and extracted with ethyl acetate (100 mL×2). The combined organic phase was washed with saturated sodium chloride solution (25 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 1/0 ~ 1/2) to obtain 7-4. MS(ESI) m/z: 467.0 [M+H ⁺ ].

Step D: Dissolve compound 7-4 (1.5g, 3.21mmol) in ethanol (10mL) and water (2mL), then add iron powder (1.80g, 32.15mmol), ammonium chloride (859.78mg, 16.07mmol) . The resulting mixture was stirred at 90°C for 2 hours. The reaction solution was filtered and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 7-5. MS(ESI) m/z: 437.3 [M+H ⁺ ].

Step E: Dissolve compound 1-8 (500mg, 1.15mmol) in dichloromethane (10mL) and add 7-5 (330.09mg, 2.29mmol), T3P (2.91g, 4.58mmol, 2.72mL, 50% purity, ) And DIEA (888.00mg, 6.87mmol, 1.20mL,), react at 25°C for 12 hours. Water (50 mL) was added to the reaction solution and extracted with dichloromethane (25 mL×2). The combined organic phase was washed with saturated sodium chloride solution (25 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 7-6. MS(ESI) m/z: 563.1 [M+H ⁺ ].

Step F: Compound 7-6 (550mg, 977.38μmol) was dissolved in tetrahydrofuran (2mL) and water (0.5mL), and then lithium hydroxide monohydrate (82.02mg, 1.95mmol) was added to the reaction solution. The resulting mixture was Stir at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure, water (10 mL) was added, the pH was adjusted to 6 with 1 mol/L hydrochloric acid, and the mixture was extracted with ethyl acetate (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 7-7. MS(ESI) m/z: 549.1 [M+H ⁺ ].

Step G: Dissolve compound 7-7 (536.29 mg, 977.38 μmol,) in tetrahydrofuran (5 mL) and then add 7-8 (108.60 mg, 977.38 μmol), T ₃ P (1.24 g, 1.95 mmol, 50% ethyl acetate) Solution) and DIEA (505.26mg, 3.91mmol), and react at 25°C for 12 hours. Water (50 mL) was added to the reaction solution and extracted with ethyl acetate (25 mL×2). The combined organic phase was washed with saturated sodium chloride solution (25 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 7-9. MS(ESI) m/z: 642.1 [M+H ⁺ ].

Step H: Compound 7-9 (480mg, 747.89μmol) was dissolved in dichloromethane (2mL), then trifluoroacetic acid (4.26g, 37.39mmol) was added to the reaction solution, and the resulting mixture was stirred at 25°C for half an hour . The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography (column chromatography: Phenomenex Luna C18 75*30*3μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 13%-35%, 10 minutes) and purified. Compound 7. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.17(s,1H), 10.04(s,1H), 8.38(d,J=5.6Hz,1H), 7.97(s,1H), 7.77(s, 1H), 7.73 (d, J = 8.8 Hz, 2H), 7.69-7.61 (m, 2H), 7.50 (d, J = 16.0 Hz, 1H), 7.22-7.07 (m, 4H), 6.90 (d, J = 16.0Hz, 1H), 6.84 (d, J = 2.4 Hz, 1H), 6.71 (dd, J = 2.4, 5.6 Hz, 1H), 5.12-4.78 (m, 1H), 4.12 (t, J = 5.6 Hz , 2H), 3.73 (q, J = 5.2 Hz, 2H), 1.47 (s, 4H); MS (ESI) m/z: 528.1 [M+H ⁺ ].

Example 8: Compound 8

Step A: Dissolve compound 3-1 (3g, 20.41mmol) in N,N-dimethylformamide (50mL), add 8-1 (3.12g, 22.45mmol), cesium carbonate (9.98 g, 30.62 mmol) and TBAI (1.51 g, 4.08 mmol), and the resulting mixture was stirred at 90°C for 4 hours. The reaction solution was quenched by adding water (50 mL) and extracted with dichloromethane (25 mL×2). The combined organic phase was washed twice with saturated sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography (column: Waters Xbridge BEH C18 250*50mm*10μm; mobile phase: [water (0.05% ammonia v/v)-ACN]; B%: 15%-40%, 20 Min) Purification to obtain 8-2. MS(ESI) m/z: 206.8 [M+H ⁺ ].

Step B: Compound 8-2 (1g, 4.88mmol), 3-6 (1.27g, 4.88mmol), palladium acetate (109.49mg, 487.68μmol), P(o-tolyl) ₃ (1.48g, 4.88mmol) , Triethylamine ((2.47g, 24.38mmol, 3.39mL) and lithium bromide (1.27g, 14.63mmol) were dissolved in toluene (10mL), the resulting mixture was replaced with nitrogen three times and stirred at 110°C for 12 hours. In the reaction solution It was quenched by adding water (100 mL) and extracted with ethyl acetate (100 mL×2). The combined organic phases were washed twice with saturated sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was passed Separation by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1-0:1) to obtain 8-3. MS (ESI) m/z: 384.9 [M+H ⁺ ].

Step C: Compound 8-3 (230 mg, 598.40 μmol) was dissolved in water (2 mL) and ethanol (10 mL), and iron powder (334.20 mg, 5.98 mmol) and ammonium chloride (160.04 mg, 2.99) were added to the reaction system. mmol), the resulting mixture was stirred at 90°C for 3 hours. After the reaction solution was filtered through Celite, the filtrate was extracted with ethyl acetate (50mL×2). The combined organic phases were washed with saturated sodium chloride solution (50mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 8-4 . MS(ESI) m/z: 355.1 [M+H ⁺ ].

Step D: Compound 8-4 (100mg, 282.19μmol) was dissolved in tetrahydrofuran (5mL), and 1-8 (125.97mg, 564.37μmol), T3P (538.71mg, 846.56μmol, 503.47μL, 50 % Purity) and DIEA (182.35 mg, 1.41 mmol, 245.75 μL), and the resulting mixture was stirred at 25° C. for 12 hours. The reaction solution was quenched by adding water (50 mL) and extracted with dichloromethane (25 mL×2). The combined organic phase was washed twice with saturated sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by high performance liquid chromatography (column information: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: water(0.1%TFA)-ACN]; B%: 30%-40%, 7 minutes) to obtain compound 8 . ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.45(s,1H),9.97(s,1H),8.54(d,J=6.4Hz,1H),8.06(s,1H),7.92(br dd ,J=1.6,13.2Hz,1H),7.84(s,1H),7.70(br d,J=16.4Hz,1H),7.63(br dd,J=5.2,8.8Hz,2H),7.52(br s ,1H),7.42(s,1H),7.34(br s,1H),7.16(br t,J=8.8Hz,2H),7.11-7.05(m,1H),6.98(d,J=16.4Hz, 1H), 4.28(br t,J=5.2Hz,2H), 3.69(br d,J=5.2Hz,2H),3.25-3.21(m,3H),1.48(br d,J=13.2Hz,4H) ;MS(ESI)m/z:560.1[M+H ⁺ ].

Example 9: Compound 9

Step A: Dissolve compound 9-1 (4.75g, 30.21mmol), 2-2 (4.5g, 30.21mmol), potassium carbonate (8.35g, 60.41mmol) in N,N-dimethylformamide (100mL) The resulting mixture was stirred at 80°C for 12 hours. Water (300 mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (100 mL×3). The combined organic phase was washed with saturated sodium chloride solution (200 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 9-2. MS(ESI)m/z:270.1[M+H ⁺ ].

Step B: The compound 9-2 (7g, 25.96mmol), 1-2 (4.80g, 31.16mmol), tetrakis (triphenylphosphorus) palladium (3.00g, 2.60mmol,), sodium carbonate (5.50g, 51.93 mmol) was dissolved in 1,4-dioxane (120 mL) and water (24 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 84°C for 12 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (100 mL×3). The combined organic phase was washed with saturated sodium chloride solution (100 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by a silica gel column (petroleum ether: ethyl acetate = 1/0 ~ 3/5) to obtain 9-3. MS(ESI)m/z:262.1[M+H ⁺ ].

Step C: Compound 9-3 (200mg, 765.67μmol,), 9-4 (539.47mg, 1.53mmol), palladium acetate (17.19mg, 76.57μmol,), triethylamine (387.39mg, 3.83mmol, 532.86μL) ), lithium bromide (199.48mg, 2.30mmol), P(o-tolyl) ₃ (233.04mg, 765.67μmol) was dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and heated at 70 Stir at °C for 12 hours. Water (30 mL) was added to the reaction solution to quench the dilution with ethyl acetate (30 mL), filtered through Celite, and the filtrate was extracted with ethyl acetate (30 mL×3). The combined organic phase was washed with saturated sodium chloride solution (30 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 1/0 ~ 1/2) to obtain 9-5. MS(ESI) m/z: 486.1 [M+H ⁺ ].

Step D: Dissolve compound 9-5 (200mg, 411.88μmol) in ethanol (5mL) and water (1mL), then add iron powder (230.01mg, 4.12mmol), ammonium chloride (110.16mg, 2.06mmol), The resulting mixture was stirred at 90°C for 1 hour. The reaction solution was filtered and extracted with ethyl acetate (20 mL×2). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 9-6. MS(ESI) m/z: 456.1 [M+H ⁺ ].

Step E: Compound 1-8 (82.79mg, 370.94μmol) was dissolved in dichloromethane (3mL) and oxalyl chloride (181.08mg, 1.43mmol), N,N-dimethylformamide ((2.09mg , 28.53μmol), the resulting mixture was stirred for half an hour at 25°C. The crude acid chloride was obtained by concentration under reduced pressure, and then the acid chloride was dissolved in dichloromethane (3mL), and pyridine (112.85mg, 1.43mmol) was added to the solution at zero degrees. Then 9-6 (130mg, 285.34μmol) dissolved in dichloromethane (3mL) was added dropwise to the mixed solution, the resulting mixture was stirred at 0°C for ten minutes, and then at 25°C for half an hour. Water was added to the reaction solution (10 mL) The reaction was quenched and extracted with dichloromethane (10 mL×2). The combined organic phases were washed with saturated sodium chloride solution (20 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 9-7. MS (ESI) m/z: 661.2 [M+H ⁺ ].

Step F: Compound 9-7 (170 mg, 257.27 μmol) was dissolved in dichloromethane (2 mL), then trifluoroacetic acid (2 mL) was added to the reaction solution, and the resulting mixture was stirred at 25° C. for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (column information: Phenomenex luna C18 150*40*15μm; mobile phase: [water (0.1%TFA)-ACN]; ACN%: 30%-60%, 10 minutes) to obtain compound 9 The trifluoroacetate. ¹ H NMR (400MHz, DMSO-d ₆ )δ = 10.33 (s, 1H), 10.01 (s, 1H), 8.63 (d, J = 5.6 Hz, 1H), 8.07 (s, 1H), 7.87-7.77 ( m, 2H), 7.68-7.57 (m, 3H), 7.49-7.43 (m, 1H), 7.38-7.31 (m, 1H), 7.15 (t, J = 8.8 Hz, 2H), 6.94 (d, J = 6.0Hz, 1H), 6.72 (d, J = 15.6Hz, 1H), 4.13-4.09 (m, 2H), 3.71 (t, J = 5.6Hz, 2H), 1.47 (br d, J = 4.0Hz, 4H ); MS (ESI) m/z: 547.1 [M+H ⁺ ].

Example 10: Compound 10

Step A: Dissolve compound 10-1 (2g, 12.20mmol) in N,N-dimethylformamide (20mL), add 10-2 (3.88g, 24.39mmol), potassium carbonate (7.95g, 24.39 mmol), and the resulting mixture was stirred at 70°C for 12 hours. Water (20 mL) was added to the reaction solution, and it was extracted with ethyl acetate (20 mL×2). The combined organic phase was washed twice with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate=1:0-5 /1/,v/v) to get 10-3. MS(ESI)m/z:303.0[M+H ⁺ ].

Step B: Compound 10-3 (700 mg, 2.31 mmol), 1-2 (426.87 mg, 2.77 mmol), tetrakis (triphenylphosphorus) palladium (266.90 mg, 230.97 μmol), sodium carbonate (489.60 mg, 4.62 mmol) ) Was dissolved in 1,4-dioxane (10 mL) and water (2 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 85°C for 12 hours. The reaction solution was diluted with water (10 mL) and ethyl acetate (10 mL), and extracted with ethyl acetate (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product is separated by the column (

10g

Fast silica gel column, the eluent is ethyl acetate/petroleum ether=0-10%) for purification to obtain 10-4. MS(ESI)m/z:295.0[M+H ⁺ ].

Step C: Compound 10-4 (0.65g, 2.21mmol), 3-4 (1.35g, 4.41mmol), palladium acetate (49.52mg, 220.59μmol), triethylamine (1.12g, 11.03mmol), tetrabutyl Amine bromide (617.07mg, 1.91mmol), P(o-tolyl) ₃ (671.40mg, 2.21mmol), lithium bromide (574.71mg, 6.62mmol) dissolved in N,N-dimethylacetamide (10mL) The resulting mixture was replaced with nitrogen three times and stirred at 125°C for 12 hours. The reaction solution was diluted with water (10 mL) and ethyl acetate (10 mL), and extracted with ethyl acetate (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 1/0 ~ 1/1) to obtain 10-5. MS(ESI) m/z: 519.0 [M+H ⁺ ].

Step D: Dissolve compound 10-5 (270mg, 520.19μmol) in ethanol (10mL) and water (2mL), then add iron powder (290.50mg, 5.20mmol), ammonium chloride (139.13mg, 2.60mmol), The resulting mixture was stirred at 90°C for 1 hour. After the reaction solution was filtered, the filtrate was extracted with ethyl acetate (20 mL×2). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 10-6. MS(ESI) m/z: 489.2 [M+H ⁺ ].

Step E: Compound 1-8 (94.93mg, 425.31μmol) was dissolved in dichloromethane (5mL), and N,N-dimethylformamide (2.39mg, 32.72μmol) was added to the reaction. Oxalyl chloride (207.63 mg, 1.64 mmol) was added to the reaction system, and the resulting mixture was stirred at 25°C for 0.5 hours. Concentrate under reduced pressure to obtain crude acid chloride. The crude acid chloride was dissolved in dichloromethane (5 mL), and the temperature was reduced to 0°C. At this temperature, pyridine (129.39 mg, 1.64 mmol) was added to the reaction system, and then 10 was added dropwise to the reaction. -6 (160mg, 327.16μmol) in dichloromethane (1mL) solution, the reaction system after dripping was stirred at 0°C for 10 minutes, and then heated to 25°C and stirred for 30 minutes. The reaction solution was diluted with water (10mL) and dichloromethane (20mL), and extracted with dichloromethane (20mL×3). The combined organic phases were washed with saturated sodium chloride solution (20mL), dried over anhydrous sodium sulfate, filtered and then depressurized Concentrate to obtain the crude product, which is separated by reversed-phase column (

10g

Fast silica gel column, the eluent is ethyl acetate/petroleum ether=0-50%) for purification to obtain 10-7. MS(ESI) m/z: 694.4 [M+H ⁺ ].

Step F: Compound 10-7 (100 mg, 144.04 μmol) was dissolved in dichloromethane (5 mL), then trifluoroacetic acid (1 mL) was added to the reaction solution, and the resulting mixture was stirred at 25° C. for 1 hour. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (column information: Phenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (0.1%TFA)-ACN]; B%: 42%-52%, 7 minutes) and purified. Compound 10 trifluoroacetate salt. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.40(d,J=2.4Hz,1H), 10.00(s,1H), 8.47-8.31(m,1H), 8.18(d,J=22.8Hz, 1H), 7.92-7.88 (m, 1H), 7.76 (d, J = 15.6Hz, 1H), 7.64 (t, J = 6.8Hz, 3H), 7.51 (br d, J = 8.8Hz, 1H), 7.43 -7.31(m,2H),7.16(t,J=8.8Hz,2H),6.77-6.67(m,1H),6.63-6.54(m,1H),4.14(td,J=5.6,16.8Hz,2H ),3.78-3.67(m,2H),1.47(br d,J=4.8Hz,4H); MS(ESI)m/z:601.1[M+H ⁺ ].

Example 11: Compound 11

Step A: Dissolve compound 11-1 (2g, 13.61mmol) in N,N-dimethylformamide (10mL), add 2-bromoethanol (1.87g, 14.97mmol, 1.06mL), carbonic acid to the reaction system Cesium (6.65g, 20.41mmol,), TBAI (1.01g, 2.72mmol), the resulting mixture was stirred at 80°C for 12 hours. The reaction solution was quenched by adding water (50 mL) and extracted with dichloromethane (20 mL×3). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude compound 11-2.

Step B: Dissolve compound 11-2 (3g, 15.70mmol) in N,N-dimethylformamide (20mL), add TBSCl (2.84g, 18.85mmol), imidazole (2.14g, 31.41) to the reaction system mmol), the resulting mixture was stirred at 25°C for 12 hours. The reaction solution was quenched by adding water (20 mL) and ethyl acetate (50 mL) and extracted with ethyl acetate (50 mL×3). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by a reverse phase column (

10g

Fast silica gel column, the eluent is 0-10% petroleum ether/ethyl acetate) to obtain 11-3. MS(ESI)m/z:305.1[M+H ⁺ ].

Step C: The compound 3-6 (1g, 3.84mmol), 11-3 (2.35g, 7.69mmol), palladium acetate (86.28mg, 384.29μmol), triethylamine (1.94g, 19.21mmol), lithium bromide (1.00 g, 11.53mmol), P(o-tolyl) ₃ (1.17g, 3.84mmol) was dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and stirred at 125°C for 12 hours . The reaction solution was diluted with water (10 mL) and ethyl acetate (10 mL), and extracted with ethyl acetate (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product is separated by a reversed-phase column (

10g

Fast silica gel column, the eluent is ethyl acetate/petroleum ether = 0-100%) for purification to obtain 11-4. MS(ESI)m/z:485.2[M+H ⁺ ].

Step D: Dissolve compound 11-4 (500mg, 1.03mmol) in ethanol (10mL) and water (2mL), then add iron powder (576.20mg, 10.32mmol), ammonium chloride (1551.91mg, 10.32mmol), The resulting mixture was stirred at 90°C for 1 hour. After the reaction solution was filtered, the filtrate was extracted with ethyl acetate (20 mL×2). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 11-5. MS(ESI)m/z:455.2[M+H ⁺ ].

Step E: Dissolve compound 1-8 (287.22mg, 1.29mmol) in dichloromethane (10mL), add N,N-dimethylformamide (7.24mg, 98.99μmol) to the reaction, and keep at 25℃ Oxalyl chloride (628.19 mg, 4.95 mmol) was added to the reaction system, and the resulting mixture was stirred at 25°C for 0.5 hours. Concentrate under reduced pressure to obtain crude acid chloride. The crude acid chloride was dissolved in dichloromethane (10 mL), and the temperature was reduced to 0°C. At this temperature, pyridine (391.49 mg, 4.95 mmol) was added to the reaction system, and then 11 was added dropwise to the reaction. -5 (450mg, 989.86μmol) in dichloromethane (1mL) solution, the reaction system after dripping was stirred at 0°C for 10 minutes, and then heated to 25°C and stirred for 30 minutes. The reaction solution was diluted with water (10 mL) and dichloromethane (20 mL), and extracted with dichloromethane (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by a chromatography plate (petroleum ether/ethyl acetate = 1/2) to obtain 11- 6. MS(ESI) m/z: 660.2 [M+H ⁺ ].

Step F: Compound 11-6 (.45g, 682.03μmol) was dissolved in dichloromethane (10mL), then trifluoroacetic acid (77.77mg, 682.03μmol) was added to the reaction solution, and the resulting mixture was stirred at 25°C for 5 Hour. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by (PPhenomenex Gemini-NX C18 75*30mm*3μm; mobile phase: [water (0.1%TFA)-ACN]; B%: 30%-40%, 7 minutes) to obtain the trifluoroacetate salt of compound 11 . ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.44 (s, 1H), 9.98 (s, 1H), 8.58 (d, J = 6.4 Hz, 1H), 7.92 (dd, J = 2.4, 13.2 Hz, 1H), 7.79-7.59 (m, 4H), 7.58-7.38 (m, 3H), 7.26-7.05 (m, 4H), 6.62 (d, J = 2.4 Hz, 1H), 4.17 (br t, J = 5.6 Hz,2H),3.75(t,J=5.6Hz,2H),1.53-1.41(m,4H); MS(ESI)m/z:546.2[M+H ⁺ ]

Example 12: Compound 12

Step A: Dissolve compound 12-1 (5g, 34.02mmol), 2-bromoethanol (4.68g, 37.42mmol), cesium carbonate (16.63g, 51.03mmol), TBAI (2.51g, 6.80mmol) in N, N -In dimethylacetamide (50 mL) and water (20 mL), the resulting mixture was stirred at 90°C for 12 hours. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (100 mL×2). The combined organic layer was washed with saturated sodium chloride solution (100 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure , Get 12-2. MS(ESI) m/z: 192.03 [M+H ⁺ ].

Step B: Dissolve compound 12-2 (6.5g, 34.03mmol), TBSCl (6.15g, 40.83mmol), imidazole (4.63g, 68.05mmol) in N,N-dimethylformamide (50mL), The resulting mixture was stirred at 25°C for 12 hours. The reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (100 mL×2). The combined organic layer was washed with saturated sodium chloride solution (50 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure , Get the crude product. The crude product is separated by a silica gel column (

80g

Fast silica gel column, eluent: ethyl acetate/petroleum ether=0-100%, flow rate 60mL/min) to obtain 12-3. MS(ESI) m/z: 307.0 [M+H ⁺ ].

Step C: The compound 12-3 (1g, 3.84mmol), 3-6 (2.34g, 7.68mmol), palladium acetate (86.28mg, 384.00μmol), tetrakis (triphenylphosphorus) palladium (1.17g, 3.84mmol) ), anhydrous lithium bromide (1.00g, 11.52mmol, 289.38μL), triethylamine (1.94g, 19.20mmol), dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and in Stir at 125°C for 12 hours. The reaction mixture was diluted with 300 mL of water and extracted with ethyl acetate (100 mL×2). The combined organic layer was washed with saturated sodium chloride solution (100 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain Crude. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=10/1 to 0/1) to obtain 12-4. MS(ESI) m/z: 485.3 [M+H ⁺ ].

Step D: Dissolve compound 12-4 (560mg, 1.16mmol) in ethanol (5mL) and water (1mL), then add iron powder (645.41mg, 11.56mmol), ammonium chloride (309.07mg, 5.78mmol), The resulting mixture was stirred at 90°C for 2 hours. The reaction solution was filtered and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 12-5. MS(ESI)m/z:455.2[M+H ⁺ ].

Step E: Dissolve compound 1-8 (294.58mg, 1.32mmol) in dichloromethane (5mL), add oxalyl chloride (698.01mg, 5.50mmol) and N,N-dimethylformamide ( 8.04mg, 109.98μmol), reacted at 25°C for half an hour. The solution was concentrated to obtain the corresponding acid chloride and dissolved in dichloromethane (5 mL), and 12-5 (500 mg, 1.10 mmol) and pyridine (434.99 mg, 5.50 mmol) were added to the solution at 0°C. The resulting mixture was stirred at 25°C for half an hour. Water (20 mL) was added to the reaction solution for liquid separation, and the aqueous phase was extracted with dichloromethane (30 mL×2). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 12-6, and the crude product was directly used in the next step. MS(ESI)m/z:660.3[M+H ⁺ ].

Step F: Compound 12-6 (450mg, 682.03μmol) was dissolved in dichloromethane (4mL), then trifluoroacetic acid (3.89g, 34.10mmol) was added to the reaction solution, and the resulting mixture was stirred at 25°C for 1 hour . The reaction solution was concentrated to obtain a crude product. Use high performance liquid chromatography to separate (column information: Phenomenex Luna C18 150*25mm*10μm; mobile phase: [water (0.225%FA)-ACN]; B%: 18%-48%, 9 minutes) to obtain the crude product, Continue to use supercritical fluid chromatography to separate (column information: DAICEL CHIRALCEL OD (250mm*30mm, 10μm); mobile phase: [0.1% ammonia in ethanol]; B%: 60%-60%, 3.9min; 100min) purified to obtain Compound 12. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.37(s,1H), 10.00(s,1H), 8.39(d,J=5.6Hz,1H), 8.14(s,1H), 7.87(dd, J = 2.4, 13.2 Hz, 1H), 7.69-7.61 (m, 3H), 7.52-7.46 (m, 2H), 7.42 (s, 1H), 7.37-7.30 (m, 1H), 7.16 (t, J = 8.8Hz, 2H), 7.07 (d, J = 16 Hz, 1H), 7.01 (d, J = 2.4 Hz, 1H), 6.67 (dd, J = 2.4, 5.6 Hz, 1H), 4.99 (br t, J = 5.2Hz, 1H), 4.00 (t, J = 5.6 Hz, 2H), 3.66 (q, J = 5.2 Hz, 2H), 1.47 (br d, J = 4.4 Hz, 4H); MS (ESI) m/z :546.0[M+H ⁺ ].

Example 13: Compound 13

Step A: Dissolve compound 1-8 (2.63g, 11.80mmol) in dichloromethane (15mL), add oxalyl chloride (2.50g, 19.67mmol) and N,N-dimethylformamide ( (28.75mg, 393.34μmol), reacted for half an hour at 25°C. The solution was concentrated to obtain the corresponding acid chloride and dissolved in tetrahydrofuran (15mL), and 2-2((2g, 15.73mmol) was added to the solution at 0°C. The result was obtained. The mixture was stirred at 70°C for two hours. Water (300 mL) was added to the reaction solution, the layers were separated, and the aqueous phase was extracted with dichloromethane (100 mL×2). The combined organic phases were washed with saturated sodium chloride solution (200 mL), and anhydrous It was dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain 13-7, which was directly used in the next step. MS (ESI) m/z: 333.1 [M+H ⁺ ].

Step B: Dissolve compound 13-7 (2g, 6.02mmol) in N,N-dimethylformamide (15mL), slowly add sodium hydride (262.63mg, 6.57mmol, 60% content) at 0°C, the mixture After reacting at 25°C for half an hour, compound 1-1 (719.69 mg, 5.47 mmol) was added to the reaction, and the resulting mixture was stirred at 120°C for 12 hours. The reaction mixture was diluted with water (300 mL) and then diluted with ethyl acetate (150 mL× 2) Extraction, the mixed organic layer was washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product is separated by a silica gel column (

40g

Fast silica gel column, eluent: ethyl acetate/petroleum ether = 0-100%) purified to obtain 13-6. MS(ESI) m/z: 444.2 [M+H ⁺ ].

Step C: Dissolve compound 3-4 (500mg, 1.64mmol), cuprous iodide (31.19mg, 163.78μmol), Xantphos (94.77mg, 163.78μmol), and palladium chloride (29.04mg, 163.78μmol) in triethyl The resulting mixture in the amine (50 mL) was replaced with nitrogen three times, and 13-1 (3.22 g, 32.76 mmol) was added to the resulting mixture and stirred at 65°C for 12 hours. The reaction mixture was filtered with Celite, diluted with water (200 mL), and extracted with ethyl acetate (100 mL×2). The mixed organic layer was washed with saturated sodium chloride solution (200 mL), and dried over anhydrous sodium sulfate. Filter and concentrate under reduced pressure to obtain 13-2. MS(ESI) m/z: 323.2 [M+H ⁺ ].

Step D: Compound 13-2 (2g, 6.20mmol) and potassium carbonate (1.71g, 12.40mmol) were dissolved in methanol (20mL), and the resulting mixture was stirred at 60°C for 2 hours. The reaction mixture was diluted with water (300 mL) and extracted with ethyl acetate (150 mL×2). The combined organic layer was washed with saturated sodium chloride solution (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain Crude. The crude product is separated by a silica gel column (

24g

Fast silica gel column, the eluent is ethyl acetate/petroleum ether = 0-100%) for purification to obtain 13-3.MS (ESI) m/z: 251.2 [M+H ⁺ ].

Step E: Dissolve compound 13-4 (1.570g, 6.18mmol), cuprous chloride (15.81mg, 159.74μmol), Xantphos (92.43mg, 159.74μmol) in tetrahydrofuran (5mL), and replace the resulting mixture with nitrogen Three times, add potassium tert-butoxide in tetrahydrofuran (1M, 1.60mL), then add 13-3 (400mg, 1.60mmol) and methyl iodide (453.46mg, 3.19mmol), the mixture was replaced with nitrogen once, the resulting mixture was at 25°C Stir for 12 hours. After the reaction was completed, a saturated aqueous ammonium chloride solution was added to the reaction system and stirred for one hour to quench. The mixture was diluted with water (50ml), extracted with ethyl acetate (100ml), the combined organic layer was washed with saturated sodium chloride solution (50mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product . The crude product was purified by thin layer chromatography to obtain 13-5. MS(ESI) m/z: 393.3 [M+H ⁺ ].

Step F: Compound 13-5 (130 mg, 331.28 μmol), compound 13-6 (176.44 mg, 397.54 μmol), tetrakis (triphenylphosphorus) palladium (38.28 mg, 33.13 μmol), sodium carbonate (70.23 mg, 662.56 μmol) was dissolved in 1,4-dioxane (1 mL) and water (0.4 mL), the mixture was replaced with nitrogen three times, and the resulting mixture was stirred at 85°C for 4 hours. The reaction mixture was diluted with 50 mL of water and extracted with 100 mL of ethyl acetate (50 mL×2). The combined organic layer was washed with saturated sodium chloride solution (25 mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. , Get the crude product. The crude product was purified by thin layer chromatography to obtain 13-9. MS(ESI) m/z: 674.3 [M+H ⁺ ].

Step G: Compound 13-9 (100mg, 148.41μmol) was dissolved in dichloromethane (1mL), and trifluoroacetic acid (846.07mg, 7.42mmol) was added to the reaction system. The resulting mixture was stirred at 25°C for 0.5 hours. The reaction was quenched by adding aqueous sodium bicarbonate solution (30 mL) to the solution and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by high performance liquid chromatography (column information: water Xbridge BEH C18 250*50mm*10μm; mobile phase: [water (0.05% ammonia)-ACN]; ACN%: 40%-60%, 20 minutes) to obtain the compound 13. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.38(s,1H),10.01(s,1H),8.45(d,J=5.6Hz,1H),7.96(s,1H),7.87(dd, J = 2.4, 13.2 Hz, 1H), 7.78 (s, 1H), 7.67-7.61 (m, 2H), 7.51-7.46 (m, 1H), 7.34 (t, J = 8.8 Hz, 1H), 7.16 (t ,J = 8.8Hz, 2H), 6.81 (dd, J = 1.6, 5.3 Hz, 2H), 6.74 (dd, J = 2.4, 5.7 Hz, 1H), 4.92 (br s, 1H), 4.11 (t, J = 5.6 Hz, 2H), 3.73 (t, J = 5.6 Hz, 2H), 2.45 (d, J = 1.2 Hz, 3H), 1.52-1.42 (m, 4H). MS(ESI)m/z:560.1[M+H ⁺ ].

Example 14: Compound 14

Step A: Compound 2-4 (855.62mg, 3.28mmol), 3-4 (1g, 3.28mmol), palladium acetate (73.54mg, 327.56μmol), triethylamine (1.66g, 16.38mmol), P(o -tolyl) ₃ (996.98 mg, 3.28 mmol), lithium bromide (853.41 mg, 9.83 mmol) was dissolved in toluene (30 mL), the resulting mixture was replaced with nitrogen three times and stirred at 110°C for 12 hours. Water (100 mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=10/1～0/1) to obtain 14-1. MS(ESI) m/z: 486.3 [M+H ⁺ ].

Step B: Dissolve compound 14-1 (300mg, 617.81μmol) in ethanol (10mL) and water (2mL), then add iron powder (345.02mg, 6.18mmol), ammonium chloride (165.24mg, 3.09mmol), The resulting mixture was stirred at 90°C for 6 hours. The reaction solution was filtered and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 14-2. MS(ESI) m/z: 456.1 [M+H ⁺ ].

Step C: Dissolve compound 14-2 (100 mg, 219.49 μmol) in dichloromethane (5 mL), and then add 1-8 (97.98 mg, 438.98 μmol), T ₃ P (419.03 mg, 658.47 μmol), DIEA ( 141.84mg, 1.10mmol). The mixture was stirred at 25°C for 12 hours. The reaction solution was quenched by adding water (50 mL) and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 14-3. MS(ESI) m/z: 661.1 [M+H ⁺ ].

Step D: Dissolve compound 14-3 (30 mg, 45.40 μmol) in dichloromethane (2 mL), add trifluoroacetic acid (709.19 mg, 6.22 mmol) to the reaction system, and stir the resulting mixture at 25°C for 1 hour. The reaction mixture Concentrate under reduced pressure to obtain crude product. The crude product was separated by high performance liquid chromatography (column information: Xtimate C18 150*40mm*10μm; mobile phase: [water (0.05% ammonia)-ACN]; ACN%: 32%-62%, 10 minutes) to obtain compound 14 . ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.32 (s, 1H), 10.02 (s, 1H), 8.62 (s, 1H), 8.09 (s, 1H), 7.87-7.77 (m, 3H), 7.67-7.62 (m, 2H), 7.44 (dd, J = 1.2, 8.8 Hz, 1H), 7.33 (t, J = 8.8 Hz, 1H), 7.16 (t, J = 8.8 Hz, 2H), 7.10 (s ,1H),6.97(d,J=16.0Hz,1H),4.94(t,J=5.2Hz,1H),4.16(t,J=5.6Hz,2H),3.74(q,J=5.2Hz,2H ),1.47(br d,J=2.8Hz,4H); MS(ESI)m/z:547.1[M+H ⁺ ].

Example 15: Compound 15

Step A: Dissolve compound 15-1 (3g, 23.16mmol), compound 15-2 (4.07g, 23.16mmol), potassium carbonate (6.40g, 46.32mmol) in N,N-dimethylformamide (20mL) The resulting mixture was stirred at 100°C for 12 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL×2). The mixed organic layer was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain Crude product, crude product is separated by silica gel column (

120g

Fast silica gel column, the eluent is ethyl acetate/petroleum ether = 0-100%) to obtain 15-3. MS (ESI) m/z: 284.7 [M+H ⁺ ].

Step B: Combine compound 15-3 (3g, 10.52mmol), 1-2 (1.94g, 12.63mmol), tetrakis (triphenylphosphorus) palladium (1.22g, 1.05mmol), sodium carbonate (2.23g, 21.05mmol) ) Was dissolved in 1,4-dioxane (30 mL) and water (6 mL), and the resulting mixture was replaced with nitrogen three times and stirred at 84°C for 4 hours. The reaction solution was quenched by adding water (100 mL) and extracted with ethyl acetate (100 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 20/1 to 5/1) to obtain 15-4. MS(ESI) m/z: 277.0 [M+H ⁺ ].

Step C: Compound 15-4 (906.28mg, 3.28mmol), 3-4 (1g, 3.28mmol), palladium acetate (73.54mg, 327.56μmol), triethylamine (1.66g, 16.38mmol), P(o -tolyl) ₃ (997.00 mg, 3.28 mmol), lithium bromide (853.46 mg, 9.83 mmol) was dissolved in toluene (10 mL), the resulting mixture was replaced with nitrogen three times and stirred at 110°C for 12 hours. Water (100 mL) was added to the reaction solution to quench the reaction and extracted with ethyl acetate (100 mL×2). The combined organic phase was washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=10/1～0/1) to obtain 15-5. MS(ESI) m/z: 501.2 [M+H ⁺ ].

Step D: Dissolve compound 15-5 (400mg, 798.33μmol) in ethanol (10mL) and water (2mL), then add iron powder (445.86mg, 7.98mmol), ammonium chloride (213.51mg, 3.99mmol), The resulting mixture was stirred at 90°C for 6 hours. The reaction solution was filtered and extracted with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude product 15-6. MS(ESI) m/z: 471.2 [M+H ⁺ ].

Step E: Compound 1-8 (56.86mg, 254.74μmol) was dissolved in dichloromethane (5mL), and oxalyl chloride (134.73mg, 1.06mmol) and N,N-dimethylformamide ( 1.55mg, 21.23μmol), reacted at 25°C for half an hour. The solution was concentrated to obtain the corresponding acid chloride and dissolved in dichloromethane (10 mL), and 15-6 (100 mg, 212.28 μmol) and pyridine (83.96 mg, 1.06 mmol) were added to the solution at 0°C. The resulting mixture was stirred at 25°C for half an hour. Water (20 mL) was added to the reaction solution for liquid separation, and the aqueous phase was extracted with dichloromethane (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 15-7, and the crude product was directly used in the next step. MS(ESI)m/z:676.3[M+H ⁺ ].

Step F: Dissolve compound 15-7 (120mg, 177.45μmol) in dichloromethane (2mL), add trifluoroacetic acid (2.77g, 24.31mmol) to the reaction system, and stir the resulting mixture at 25°C for 1 hour. The reaction mixture Concentrate under reduced pressure to obtain crude product. The crude product was separated by high performance liquid chromatography (column information: Xtimate C18 150*40mm*10μm; mobile phase: [water (0.05% ammonia)-ACN]; ACN%: 37%-67%, 10 minutes) to obtain compound 15 . ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.34(s,1H),10.01(s,1H),8.40(d,J=5.6Hz,1H), 8.06(d,J=2.4Hz,1H) ,7.97(s,1H),7.77(s,1H),7.68-7.62(m,3H),7.52(d,J=16Hz,1H),7.35(d,J=8.8Hz,1H),7.16(t ,J=8.8Hz,2H), 6.92(d,J=16Hz,1H), 6.81(d,J=2.4Hz,1H), 6.69(dd,J=2.4,5.6Hz,1H), 4.91(br s ,1H), 4.12 (t, J = 5.6 Hz, 2H), 3.73 (br d, J = 4.4 Hz, 2H), 1.47 (br d, J = 5.6 Hz, 4H); MS (ESI) m/z: 562.4[M+H ⁺ ].

Example 16: Compound 16

Step A: Dissolve compound 6 (300mg, 598.23μmol), 16-1 (239.80mg, 1.79mmol), cesium carbonate (584.74mg, 1.79mmol), potassium iodide (9.93mg, 59.82μmol) in N,N-dimethyl In methylacetamide (2 mL), the resulting mixture was stirred at 100°C for 12 hours. The reaction mixture was diluted with water (10 mL) and ethyl acetate (20 mL), separated, the aqueous phase was extracted with ethyl acetate (20 mL×2), the mixed organic layer was washed with saturated brine (20 mL×2), and anhydrous sulfuric acid It was dried with sodium, filtered, and concentrated under reduced pressure to obtain the crude product, which was separated by preparative thin layer chromatography (dichloromethane/methanol=5/1) to obtain compound 16. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.40(s,1H), 10.01(s,1H), 8.39(d,J=5.6Hz,1H), 8.07(s,1H), 7.87(dd, J = 2.2, 13.2 Hz, 1H), 7.78 (s, 1H), 7.67-7.60 (m, 2H), 7.54-7.45 (m, 2H), 7.39-7.31 (m, 1H), 7.20-7.12 (m, 2H), 6.92 (d, J = 16.0 Hz, 1H), 6.83 (d, J = 2.4 Hz, 1H), 6.74 (dd, J = 2.4, 5.6 Hz, 1H), 4.16-4.05 (m, 1H), 2.88 (br d, J = 11.2Hz, 2H), 2.24 (s, 3H), 2.17-2.04 (m, 2H), 2.03-1.89 (m, 4H), 1.51-1.41 (m, 4H); MS (ESI ) m/z: 599.4 [M+H ⁺ ].

Example 17: Compound 17

Step A: Dissolve compound 6 (50mg, 99.70μmol), 17-1 (33.28mg, 119.64μmol), cesium carbonate (97.46mg, 299.10μmol) in N,N-dimethylacetamide (2mL), The resulting mixture was stirred at 60°C for 2 hours. The reaction mixture was diluted with water (10 mL) and ethyl acetate (20 mL), separated, the aqueous phase was extracted with ethyl acetate (20 mL×2), the mixed organic layer was washed with saturated brine (20 mL×2), and anhydrous sulfuric acid It was dried with sodium, filtered, and concentrated under reduced pressure to obtain a crude product, which was separated by preparative thin-layer chromatography (dichloromethane/methanol=5/1) to obtain compound 17-2. MS (ESI) m/z: 699.2 [M+H ⁺ ].

Step B: Dissolve compound 17-2 (70mg, 100.18μmol) in ethyl acetate (3mL), add hydrochloric acid methanol solution (4mol/L, 3mL) to the reaction system, and stir the mixture for 30 minutes at 20°C. The reaction mixture Concentrate under reduced pressure to obtain crude product. The crude product is separated by high performance liquid chromatography (column information: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225%FA)-ACN]; B%: 15%-35%, 10 minutes) Purification provides compound 17-3. MS (ESI) m/z: 599.3 [M+H ⁺ ].

Step C: Compound 17-3 (97.71 mg, 163.22 μmol) was dissolved in tetrahydrofuran (10 mL), and 37 aqueous formaldehyde solution (49.01 mg, 1.63 mmol) and sodium triacetyl borohydride (172.97 mg, 816.10 μmol) were added to obtain a mixture Stir at 20°C for 12 hours. The reaction was quenched with saturated sodium bicarbonate (5mL), diluted with water (5mL), separated, the aqueous phase was extracted with ethyl acetate (20mL×3), the mixed organic layer was washed with saturated brine (20mL), anhydrous Dry over sodium sulfate, filter, and concentrate under reduced pressure to obtain the crude product. The crude product is separated by preparative high performance liquid chromatography (column information: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225％FA)- ACN]; B%: 15%-35%, 10 minutes) to obtain the formate salt of compound 17. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.41 (s, 1H), 10.02 (s, 1H), 8.40 (d, J = 5.6 Hz, 1H), 8.27 (s, 1H), 7.98 (s, 1H), 7.88(dd,J=2.4,13.2Hz,1H),7.78(s,1H),7.65(dd,J=5.2,9.2Hz,2H),7.55-7.45(m,2H),7.35(t ,J=9.2Hz,1H), 7.16(t,J=8.8Hz,2H), 6.92(d,J=16.0Hz,1H), 6.86(d,J=2.4Hz,1H), 6.74(dd,J =2.4,5.6Hz,1H),3.99(br d,J=7.2Hz,2H), 2.92(br d,J=11.2Hz,2H), 2.30(s,3H), 2.12(br t,J=11.2 Hz, 2H), 1.83 (br dd, J = 3.6, 6.8 Hz, 1H), 1.57-1.41 (m, 6H), 1.28 (q, J = 10.8 Hz, 2H); MS (ESI) m/z: 613.3 [M+H ⁺ ].

Example 18: Compound 18

Step A: Dissolve compound 6 (50mg, 99.70μmol), 18-1 (34.96mg, 119.65μmol), cesium carbonate (97.46mg, 299.10μmol) in N,N-dimethylacetamide (2mL), The resulting mixture was stirred at 60°C for 2 hours. The reaction mixture was diluted with water (10 mL) and ethyl acetate (20 mL), separated, the aqueous phase was extracted with ethyl acetate (20 mL×2), the mixed organic layer was washed with saturated brine (20 mL×2), and anhydrous sulfuric acid It was dried with sodium, filtered, and concentrated under reduced pressure to obtain the crude product, which was separated by preparative thin layer chromatography (dichloromethane/methanol=5/1) to obtain compound 18-2. MS(ESI) m/z: 713.2 [M+H ⁺ ].

Step B: Dissolve compound 18-2 (70 mg, 98.21 μmol) in ethyl acetate (3 mL), add hydrochloric acid methanol solution (4 mol/L, 3 mL) to the reaction system, and stir the mixture for 30 minutes at 20°C. The reaction mixture Concentrate under reduced pressure to obtain crude product. The crude product was separated by preparative high performance liquid chromatography (column information: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225％FA)-ACN]; ACN%: 15%-35%, 10 Min) Purification to obtain compound 18-3. MS(ESI) m/z: 613.3 [M+H ⁺ ].

Step C: Compound 18-3 (100mg, 163.22μmol) was dissolved in tetrahydrofuran (10mL), 37 formaldehyde aqueous solution (49.01mg, 1.63mmol) and sodium triacetyl borohydride (172.97mg, 816.10μmol) were added, and the resulting mixture was Stir at 20°C for 12 hours. The reaction was quenched with saturated sodium bicarbonate (5mL), diluted with water (5mL), separated, the aqueous phase was extracted with ethyl acetate (20mL×3), the mixed organic layer was washed with saturated brine (20mL), anhydrous Dry over sodium sulfate, filter, and concentrate under reduced pressure to obtain the crude product. The crude product is separated by preparative high performance liquid chromatography (column information: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225％FA)- ACN]; ACN%: 15%-35%, 10 minutes) to obtain the formate salt of compound 18. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.42 (br s, 1H), 10.03 (s, 1H), 8.40 (d, J = 5.6 Hz, 1H), 8.31 (s, 1H), 8.01 (s ,1H),7.88(dd,J=2.3,13.2Hz,1H),7.77(s,1H),7.68-7.62(m,2H),7.54-7.47(m,2H),7.35(t,J=9.0 Hz, 1H), 7.16 (t, J = 9.0 Hz, 2H), 6.92 (d, J = 15.9 Hz, 1H), 6.86 (d, J = 2.4 Hz, 1H), 6.74 (dd, J = 2.4, 5.6 Hz, 1H), 4.12 (br t, J = 7.1 Hz, 2H), 2.93 (br d, J = 11.7 Hz, 2H), 2.31 (s, 3H), 2.16 (br t, J = 10.9 Hz, 2H) , 1.79-1.60 (m, 4H), 1.54-1.39 (m, 4H), 1.33-1.08 (m, 3H); MS (ESI) m/z: 627.3 [M+H ⁺ ].

Example 19: Compound 19

Step A: Dissolve compound 6 (50mg, 99.70μmol), 19-1 (35.08mg, 119.64μmol), cesium carbonate (97.46mg, 299.10μmol) in N,N-dimethylacetamide (2mL), The resulting mixture was stirred at 60°C for 2 hours. The reaction mixture was diluted with water (10 mL) and ethyl acetate (20 mL), separated, the aqueous phase was extracted with ethyl acetate (20 mL×2), the mixed organic layer was washed with saturated brine (20 mL×2), and anhydrous sulfuric acid It was dried with sodium, filtered, and concentrated under reduced pressure to obtain the crude product, which was separated by preparative thin layer chromatography (dichloromethane/methanol=5/1) to obtain compound 19-2. MS(ESI) m/z: 714.2 [M+H ⁺ ].

Step B: Dissolve compound 19-2 (70mg, 98.07μmol) in ethyl acetate (3mL), add hydrochloric acid methanol solution (4mol/L, 3mL) to the reaction system, and stir the mixture for 30 minutes at 20°C. The reaction mixture Concentrate under reduced pressure to obtain crude product. The crude product is separated by high performance liquid chromatography (column information: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225%FA)-ACN]; B%: 15%-35%, 10 minutes) Purification afforded compound 19-3. MS(ESI) m/z: 614.3 [M+H ⁺ ].

Step C: Compound 19-3 (100 mg, 162.96 μmol) was dissolved in tetrahydrofuran (10 mL), and 37% aqueous formaldehyde solution (48.94 mg, 1.63 mmol) and sodium triacetyl borohydride (172.69 mg, 814.79 μmol) were added to obtain a mixture Stir at 20°C for 12 hours. The reaction was quenched with saturated sodium bicarbonate (5mL), diluted with water (5mL), separated, the aqueous phase was extracted with ethyl acetate (20mL×3), the mixed organic layer was washed with saturated brine (20mL), anhydrous Dry over sodium sulfate, filter, and concentrate under reduced pressure to obtain the crude product, which is separated by high performance liquid chromatography (column information: Unisil 3-100 C18 Ultra 150*50mm*3μm; mobile phase: [water (0.225% FA)-ACN] ; B%: 15%-35%, 10 minutes) to obtain the formate salt of compound 19. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.40 (s, 1H), 10.02 (s, 1H), 8.40 (d, J = 5.6 Hz, 1H), 8.23 (s, 1H), 7.99 (s, 1H), 7.88(dd,J=2.0,13.2Hz,1H),7.77(s,1H),7.65(dd,J=5.2,8.8Hz,2H),7.56-7.45(m,2H),7.35(t ,J=9.2Hz,1H), 7.16(t,J=8.8Hz,2H), 6.91(d,J=16.0Hz,1H), 6.87(d,J=2.0Hz,1H), 6.74(dd,J =2.4,5.6Hz,1H), 4.19(br t,J=6.4Hz,2H), 2.70(br t,J=6.4Hz,2H), 2.50-2.31(m,8H), 2.20(s,3H) , 1.57-1.39 (m, 4H); MS (ESI) m/z: 628.3 [M+H ⁺ ].

Example 20: Compound 20

Step A: Dissolve compound 20-1 (5.08g, 19.10mmol), compound 20-2 (5.02g, 21.00mmol), cesium carbonate (12.44g, 38.19mmol) in N,N-dimethylacetamide (50mL In), the resulting mixture was stirred at 60°C for 12 hours. The reaction mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL×2). The mixed organic layer was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. Using silica gel Column separation (petroleum ether/ethyl acetate=1:0～5:1) was purified to obtain compound 20-3. MS(ESI) m/z: 425.3 [M+H ⁺ ].

Step B: Compound 20-3 (2.4g, 5.66mmol) was dissolved in ethanol (20mL), sodium borohydride (641.91mg, 16.97mmol) was added to the reaction system, the resulting mixture was stirred at 90°C for 12 hours, and hydrochloric acid was added (1mol/L, 10mL) The reaction was quenched, water (50mL) was added, and the mixture was extracted with ethyl acetate (50mL×2). The mixed organic layer was washed with saturated brine (50mL), dried with anhydrous sodium sulfate, and filtered. Concentrate under reduced pressure to obtain compound 20-4. MS(ESI) m/z: 383.3 [M+H ⁺ ].

Step C: Dissolve compound 20-4 (2.9g, 7.59mmol) in N,N-dimethylacetamide (15mL), add TBSCl (1.37g, 9.10mmol) and imidazole (1.03g, 15.17mmol) to obtain The mixture was stirred at 25°C for 12 hours. Dilute with water (50mL) and extract with ethyl acetate (50mL×2). The mixed organic layer was washed with saturated brine (80mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. Separation by high performance liquid chromatography (column information: Phenomenex luna C18 (250*70mm, 15μm); mobile phase: [water (0.1%TFA)-ACN]; B%: 70ACN%-100ACN%, 30 minutes) purification to obtain the compound 20-5. MS(ESI) m/z: 497.4 [M+H ⁺ ].

Step D: Combine compound 3-6 (655.04mg, 2.52mmol), compound 20-5 (1.25g, 2.52mmol), palladium acetate (56.51mg, 251.73μmol), triethylamine (1.27g, 12.59mmol), lithium bromide (655.87mg, 7.55mmol), P(o-tolyl) ₃ (766.18mg, 2.52mmol) was dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and stirred at 110°C 12 hours. Water (50 mL) was added to the reaction solution and extracted with ethyl acetate (50 mL×2). The combined organic phases were washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=5:1) to obtain 20-6. MS(ESI)m/z:629.5[M+H ⁺ ].

Step E: Dissolve compound 20-6 (1.5g, 2.39mmol) in ethanol (15mL), then add iron powder (1.33g, 23.85mmol), ammonium chloride solution (127.59mg, 2.39mmol, 2.5mL water), The resulting mixture was stirred at 80°C for 12 hours. Filter, collect the filtrate, add water (50 mL), and extract with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 20-7, and the crude product was directly used in the next step. MS(ESI)m/z:599.5[M+H ⁺ ].

Step F: Dissolve compound 1-8 (339.14mg, 1.52mmol) in dichloromethane (10mL), add oxalyl chloride (741.79mg, 5.84mmol) and N,N-dimethylformamide ( 8μL), react at 25°C for half an hour. The solution was concentrated to obtain the corresponding acid chloride and dissolved in dichloromethane (10 mL), 20-7 (700 mg, 1.17 mmol) and pyridine (462.27 mg, 5.84 mmol) were added to the solution at 0°C. The resulting mixture was stirred at 25°C half an hour. Water (30 mL) was added to the reaction solution for liquid separation, and the aqueous phase was extracted with dichloromethane (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 20-8, and the crude product was directly used in the next step. MS(ESI)m/z:804.2[M+H ⁺ ].

Step G: Dissolve compound 20-8 (0.9g, 1.12mmol) in dichloromethane (20mL), then add hydrochloric acid methanol solution (4mol/L, 5mL) and ethyl acetate (5mL) to the reaction solution to obtain The mixture was stirred at 25°C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was separated by preparative high performance liquid chromatography (column information: Phenomenex luna C18 150*40mm*15μm; mobile phase: [water (0.225% FA)-ACN]; ACN%: 15%-45%, 10 minutes) and purified. Compound 20. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.36 (s, 1H), 9.99 (s, 1H), 8.39 (d, J = 5.6 Hz, 1H), 8.00 (s, 1H), 7.86 (dd, J = 2.4, 13.2 Hz, 1H), 7.66-7.59 (m, 3H), 7.48 (dd, J = 1.2, 8.8 Hz, 1H), 7.38-7.31 (m, 1H), 7.15 (t, J = 8.8 Hz ,2H),6.91-6.84(m,2H),6.71(dd,J=2.4,5.6Hz,1H),5.08(t,J=5.2Hz,1H),4.91(t,J=5.2Hz,1H) ,4.49(d,J=5.2Hz,2H),4.07(t,J=5.6Hz,2H),3.72(q,J=5.6Hz,2H),1.50-1.43(m,4H); MS(ESI) m/z:576.3[M+H ⁺ ].

Example 21: Compound 21

Step A: Compound 3-6 (735.87mg, 2.83mmol), compound 20-3 (1.2g, 2.83mmol), palladium acetate (63.49mg, 283.00μmol), triethylamine (1.43g, 14.15mmol), lithium bromide (736.80mg, 8.49mmol), P(o-tolyl) ₃ (860.72mg, 2.83mmol) was dissolved in N,N-dimethylacetamide (10mL), the resulting mixture was replaced with nitrogen three times and stirred at 110°C 12 hours. Water (50 mL) was added to the reaction solution and extracted with ethyl acetate (50 mL×2). The combined organic phases were washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated and purified by silica gel column (petroleum ether/ethyl acetate=1:1-0:1) to obtain 21-1. MS(ESI) m/z: 557.4 [M+H ⁺ ].

Step B: Dissolve compound 21-1 (1.36g, 2.44mmol) in ethanol (25mL), then add iron powder (1.21g, 21.60mmol), ammonium chloride solution (1.34g, 24.97mmol, 2.5mL water), The resulting mixture was stirred at 80°C for 12 hours. Filter, collect the filtrate, add water (50 mL), and extract with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 21-2, and the crude product was directly used in the next step. MS(ESI) m/z: 527.4 [M+H ⁺ ].

Step C: Dissolve compound 1-8 (330.56 mg, 1.48 mmol) in dichloromethane (10 mL), add oxalyl chloride (723.01 mg, 5.70 mmol) and N,N-dimethylformamide ( 8μL), react at 25°C for half an hour. The solution was concentrated to obtain the corresponding acid chloride and dissolved in dichloromethane (10 mL). 21-2 (600 mg, 1.14 mmol) and pyridine (450.56 mg, 5.70 mmol) were added to the solution at 0°C. The resulting mixture was stirred at 25°C half an hour. Water (30 mL) was added to the reaction solution for liquid separation, and the aqueous phase was extracted with dichloromethane (20 mL×3). The combined organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 21-3, and the crude product was directly used in the next step. MS(ESI) m/z: 732.3 [M+H ⁺ ].

Step D: Compound 21-3 (1g, 1.37mmol) was dissolved in methanol (5mL) and water (5mL), and then lithium hydroxide (65.45mg, 2.73mmol) was added to the reaction solution. The resulting mixture was kept at 25°C. Stir for 18 hours. Use hydrochloric acid (1mol/L) to adjust the pH to about 3, add water (50mL), and extract with ethyl acetate (50mL×2). The combined organic phase was washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain 21-4, and the crude product was directly used in the next step. MS(ESI) m/z: 590.2 [M+H ⁺ ].

Step E: Dissolve compound 21-4 (100mg, 169.62μmol) in N,N-dimethylacetamide (2mL), add ammonium chloride (18.15mg, 339.24μmol), PyBOP (105.92mg, 203.55μmol) , DIEA (87.69 mg, 678.49 μmol) and HOBt (27.50 mg, 203.55 μmol), and the resulting mixture was stirred at 25°C for 12 hours. Water (20 mL) was added, and extraction was performed with ethyl acetate (20 mL×2). The combined organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was separated by high performance liquid chromatography (column information: Phenomenex Luna C18 150*25mm*10μm; mobile phase : [water (0.225% FA)-ACN]; ACN%: 22%-52%, 11 minutes) to obtain compound 21. ¹ H NMR(400MHz,DMSO-d ₆ )δ=10.39(s,1H),10.02(s,1H), 8.42(d,J=5.6Hz,1H), 8.23(s,1H), 8.12(d, J = 16.4 Hz, 1H), 7.87 (dd, J = 2.4, 13.2 Hz, 1H), 7.68-7.60 (m, 2H), 7.49 (dd, J = 1.2, 8.8 Hz, 1H), 7.43 (s, 1H) ), 7.35(t,J=9.2Hz,1H),7.26(s,1H),7.19-7.11(m,2H),7.00(d,J=16.4Hz,1H),6.80(d,J=2.4Hz ,1H), 6.76(dd,J=2.4,5.6Hz,1H), 4.16(t,J=5.2Hz,2H), 3.78(t,J=5.2Hz,2H),1.49–1.41(m,4H) ;MS(ESI)m/z:589.4[M+H ⁺ ].

Example 22: Compound 22

Step A: Dissolve compound 21-4 (380mg, 644.56μmol) in acetonitrile (9mL), add methylamine hydrochloride (87.04mg, 1.29mmol), HATU (269.59mg, 709.02μmol), DIEA (333.21mg, 2.58 mmol) and N,N-dimethylacetamide (0.9 mL), and the resulting mixture was stirred at 25°C for 12 hours. Water (50 mL) was added, and extraction was performed with dichloromethane (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was separated by high performance liquid chromatography (column information: Waters Xbridge 150*25mm*5μm; mobile phase: [water(10mM NH4HCO3)-ACN]; ACN%: 27%-57%, 9 minutes) purified to obtain compound 22.1H NMR(400MHz,DMSO-d6)δ=10.36(s,1H),9.99(s,1H ), 8.42 (d, J = 5.6 Hz, 1H), 8.23 (s, 1H), 8.11 (d, J = 16.4 Hz, 1H), 8.05 (d, J = 4.8 Hz, 1H), 7.86 (dd, J ＝2.4,13.2Hz,1H),7.67-7.60(m,2H),7.49(dd,J＝1.2,8.8Hz,1H),7.38-7.32(m,1H),7.19-7.11(m,2H), 7.00 (d, J = 16.4 Hz, 1H), 6.81 (d, J = 2.4 Hz, 1H), 6.75 (dd, J = 2.4, 5.6 Hz, 1H), 4.99 (t, J = 5.2 Hz, 1H), 4.16(t,J=5.6Hz,2H), 3.78(q,J=4.8Hz,2H), 2.74(d,J=4.8Hz,3H), 1.49-1.43(m,4H); MS(ESI)m /z:603.2[M+H+].

Example 23: Compound 23

Step A: Dissolve compound 21-4 (100mg, 169.62μmol) in N,N-dimethylacetamide (2mL), add hydroxylamine hydrochloride (23.57mg, 339.24μmol), PyBOP (105.92mg, 203.55μmol), DIEA (109.61 mg, 848.11 μmol) and HOBt (27.50 mg, 203.55 μmol), the resulting mixture was stirred at 25°C for 12 hours. Water (20 mL) was added, and extraction was performed with ethyl acetate (20 mL×2). The combined organic phase was washed with saturated sodium chloride solution (30mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was separated by high performance liquid chromatography (column information: Unisil 3-100 C18 Ultra 150*50mm*3μm ; Mobile phase: [water (0.225% FA)-ACN]; ACN%: 25%-45%, 10 minutes) to obtain compound 23. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.88 (s, 1H), 10.37 (s, 1H), 10.00 (s, 1H), 8.94 (d, J = 1.2 Hz, 1H), 8.43 (d, J = 5.6Hz, 1H), 8.23 (s, 1H), 7.99 (d, J = 16.4 Hz, 1H), 7.87 (dd, J = 2.4, 13.2 Hz, 1H), 7.70-7.60 (m, 2H), 7.49 (dd, J = 1.6, 8.8 Hz, 1H), 7.40-7.32 (m, 1H), 7.16 (t, J = 9.2 Hz, 2H), 7.02 (d, J = 16.4 Hz, 1H), 6.83-6.73 (m, 2H), 4.98 (t, J = 5.6 Hz, 1H), 4.16 (t, J = 5.2 Hz, 2H), 3.78 (q, J = 5.2 Hz, 2H), 1.51-1.44 (m, 4H) ;MS(ESI)m/z:605.4[M+H ⁺ ].

Example 24: Compound 24

Step A: Dissolve compound 21-4 (100mg, 169.62μmol) in N,N-dimethylacetamide (2mL), add hydroxyethylamine (20.72mg, 339.24μmol), PyBOP (105.92mg, 203.55μmol) ), DIEA (109.61 mg, 848.11 μmol) and HOBt (27.50 mg, 203.55 μmol), and the resulting mixture was stirred at 25°C for 12 hours. Water (50 mL) was added, and extraction was performed with ethyl acetate (50 mL×2). The combined organic phase was washed with saturated sodium chloride solution (50mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product, which was separated by high performance liquid chromatography (column information: Phenomenex Gemini-NX C18 75*30mm*3μm; Mobile phase: [water (0.225% FA)-ACN]; ACN%: 15%-45%, 7 minutes) to obtain compound 24 for purification. ¹ H NMR (400MHz, DMSO-d ₆ ) δ = 10.42 (s, 1H), 9.99 (s, 1H), 8.50 (d, J = 6.0 Hz, 1H), 8.27 (s, 1H), 8.17 (d, J = 16.4 Hz, 1H), 7.98 (t, J = 5.8 Hz, 1H), 7.90 (dd, J = 2.4, 13.2 Hz, 1H), 7.67-7.60 (m, 2H), 7.52 (dd, J = 1.2 ,8.8Hz,1H),7.40(t,J=8.8Hz,1H),7.19-7.11(m,3H),7.09(s,1H),6.96(s,1H),4.20(t,J=5.2Hz ,2H),3.79(t,J=5.2Hz,2H),3.53-3.48(m,4H),1.52-1.42(m,4H); MS(ESI)m/z:633.1[M+H ⁺ ].

Experimental example 1: Vascular endothelial cell growth factor receptor 2 (VEGFR-2/KDR) kinase activity inhibition test (in vitro evaluation)

Experimental Materials:

KDR Kinase Enzyme System (KDR kinase enzyme system) was purchased from Promega. Nivo multi-label analyzer (PerkinElmer).

experimental method:

Use the kinase buffer in the kit to dilute the enzyme, substrate, ATP and inhibitor. The compound to be tested was diluted 5 times with a discharge gun to the 8th concentration, that is, diluted from 50 μM to 0.65 nM, the final concentration of DMSO was 5%, and the double-well experiment was set up. Add 1μL of each concentration gradient of inhibitor, 2μL of KDR enzyme (1.5ng), 2μL of a mixture of substrate and ATP (138.8μM ATP, 0.2μg/μl Poly E ₄ Y ₁ ) to the microplate, and the final concentration gradient of the compound Dilute to 0.13nM from 10μM. The reaction system was placed at 25°C for 60 minutes. After the reaction, add 5μl of ADP-Glo reagent to each well, continue the reaction at 25°C for 40 minutes, add 10μL of kinase detection reagent to each well after the reaction is over, react at 25°C for 30 minutes, use PerkinElmer Envision multi-label analyzer to read chemiluminescence, and integrate The time is 0.5 seconds.

data analysis:

Using the equation (Sample-Min)/(Max-Min)*100% to convert the original data into inhibition rate, _{the value of IC 50} can be obtained by curve fitting with four parameters (log (inhibition) vs. response in GraphPad Prism --The variable slope mode is derived).

The experimental results are shown in Table 1.

Experimental conclusion: The compound of the present invention has good inhibitory activity on VEGFR2/KDR kinase.

Experimental example 2: Axl kinase activity inhibition test (in vitro evaluation)

Experimental Materials:

AXL Kinase Enzyme System (AXL Kinase Enzyme System) was purchased from Promega. Nivo multi-label analyzer (PerkinElmer). experimental method:

Use the kinase buffer in the kit to dilute the enzyme, substrate, ATP, and inhibitor.

The compound to be tested was diluted 5-fold to the 8th concentration with a discharge gun, that is, diluted from 5 μM to 0.065 nM, with a DMSO concentration of 5%, and a double-well experiment was set up. Add 1μL of each concentration gradient of inhibitor, 2μl of AXL enzyme (6ng), 2μL of a mixture of substrate and ATP (89.37μM of ATP, 0.2μg/μL of Axltide) to the microplate, and the final concentration gradient of the compound is 1μM diluted to 0.013 nM. The reaction system was placed at 25°C for 60 minutes. After the reaction, add 5μL of ADP-Glo reagent to each well, continue the reaction at 25°C for 40 minutes, add 10μL of kinase detection reagent to each well after the reaction, react for 30 minutes at 25°C, use PerkinElmer Envision multi-label analyzer to read chemiluminescence, and integrate The time is 0.5 seconds.

data analysis:

Using the equation (Sample-Min)/(Max-Min)*100% to convert the original data into inhibition rate, _{the value of IC 50} can be obtained by curve fitting with four parameters (log (inhibition) vs. response in GraphPad Prism -The variable slope mode is derived). Table 1 provides the AXL enzymatic inhibitory activity of the compounds of the present invention.

The experimental results are shown in Table 1.

Table 1 Kinase activity inhibition test results

受试化合物Test compound	Axl(nM)Axl(nM)	VEGFR2(nM)VEGFR2(nM)
化合物1Compound 1	23.1623.16	25.8425.84
化合物2的甲酸盐Compound 2 formate	36.6236.62	44.1244.12
化合物3的甲酸盐Compound 3 formate	11.3411.34	37.3737.37
化合物4Compound 4	19.1419.14	31.5931.59
化合物6Compound 6	37.9837.98	96.4896.48
化合物7Compound 7	20.1720.17	76.9876.98
化合物8Compound 8	13.2513.25	//
化合物9的三氟乙酸盐Compound 9 trifluoroacetate	34.2334.23	74.2874.28
化合物11的三氟乙酸盐Compound 11 trifluoroacetate	33.0733.07	42.6942.69
化合物12Compound 12	11.4811.48	74.9474.94
化合物13Compound 13	97.2597.25	224.2224.2
化合物14Compound 14	25.3825.38	46.946.9
化合物15Compound 15	17.0917.09	//
化合物16Compound 16	8.58.5	17.0917.09
化合物17的甲酸盐Compound 17 formate	12.4412.44	20.5420.54
化合物18的甲酸盐Compound 18 formate	//	37.1737.17
化合物19的甲酸盐Formate of compound 19	13.8913.89	20.5120.51
化合物20Compound 20	//	33.5633.56
化合物21Compound 21	1.791.79	19.4719.47
化合物22Compound 22	4.224.22	12.7612.76
化合物23Compound 23	//	48.7148.71
化合物24Compound 24	//	14.8114.81

"/" means that the test result has not been obtained yet.

Experimental conclusion: The compound of the present invention has good inhibitory activity on AXL and VEGFR2 kinase.

Experimental example 3: n-octanol/water partition coefficient pH7.4 (Log D _7.4 )

Experimental Materials:

DMSO stock solution to prepare 10Mm DMSO stock solution of test compounds and controls (nadolol, mexetine, propranolol, quinidine, amitriptyline, chlorpromazine)

Buffer solution 100mM phosphate buffer, pH 7.4, DMSO%: 1%; n-octanol saturated buffer (pH 7.4) solution; buffer (pH 7.4) saturated n-octanol solution

Experimental operation:

1. Test compound (10mM DMSO solution, 2μL/well) and QC sample (10mM DMSO solution; 2μL/well) were added from the stock solution to a 96-well polypropylene tube;

2. Add n-octanol saturated buffer solution (150μL/well) and buffer (pH7.4) saturated n-octanol (150μL/well) solution to each hole;

3. Stir vigorously for one minute, then shake, 600 revolutions per minute, and the temperature is 28°C;

4. Centrifuge at 2500 rpm for 10 minutes

5. The n-octanol layer is diluted Y times (200 times), and the buffer layer is diluted X times (20 times)

6. Determine the test mixture concentration of the filtrate by LCMS/MS method.

The formula for calculating the Log D value of the compound is as follows:

O-S: sample peak area in the n-octanol layer

O-IS: Internal standard peak area in the n-octanol layer

B-S: sample peak area in the buffer layer

B-IS: Internal standard peak area in the buffer layer

Y: The dilution factor of the sample in the n-octanol layer

X: The dilution factor of the sample in the buffer layer

Note: The sample analysis is carried out with a three-pole quadrupole mass spectrometer. The diluted sample is injected into a small cylindrical high performance liquid chromatography column with an equal volume of water-loaded solvent, and then flushed into the mass spectrometer with a rapid gradient to obtain an organic elution solvent. The peak area is calibrated by the dilution factor, combined with the internal standard, and the result (Log D value) is calculated by the ratio of the calibrated peak area.

Experimental results: see Table 2.

Table 2 Log D _7.4 test results

受试化合物Test compound	SitravatinibSitravatinib	化合物3的甲酸盐Compound 3 formate
Log D _7.4 Log D _7.4	>4.0>4.0	3.653.65

Experimental conclusion: Log D _7.4 is the oil-water distribution coefficient of the compound at pH 7.4, which represents the physical and chemical properties of the compound. Log D _7.4 around 3 is considered to have the best druggability. Therefore, the compound of the present invention has better physical and chemical properties than Sitravatinib.

Experimental example 4: In vitro pharmacokinetic study

(1) MDR1-MDCK cell two-way permeability evaluation experiment

In this study, the MDR1-MDCKII cell line authorized by the Piet Borst Laboratory of the Netherlands Cancer Institute was used as the experimental in vitro model. It is a Madin-Darby canine kidney cell transfected with the human multi-drug resistance gene (MDR1). Cells can stably express the efflux transporter P-gp, so it is suitable for screening P-gp substrates or inhibitors, and predicts that compounds have high efflux effects in the duodenum, blood-brain barrier, hepatocyte nucleus and kidney units, etc. The permeability of the barrier. The purpose of this study is to use MDR1-MDCK II cells to determine the two-way permeability of the compound of the present invention through the MDR1-MDCK II cell model.

Experimental operation: The experimental standard conditions are as follows:

-Test concentration: 2μM (DMSO≤1%);

-Repeat: n=2;

-Direction: Two-way transfer, including two directions: A→B and B→A;

-Incubation time: a single time point, 2.5 hours;

-Transport buffer: HBSS buffer containing 10mM Hepes, pH7.4;

-Incubation conditions: 37°C, 5% CO ₂ .

After the incubation, the sample solution in the dosing hole and the receiving hole is immediately mixed with the cold acetonitrile solution containing the internal standard. The LC/MS/MS method was used to analyze the concentration of the test compound in all samples (including the initial dosing solution, the supernatant of the dosing hole, and the receiving solution). And calculate the apparent permeability coefficient, efflux ratio and other parameters.

Experimental results: Table 3 lists the permeability coefficients of Sitravatinib and compound 3 in MDR1-MDCK II monolayer cells. The results show that compound 3 has moderate permeability, while Sitravatinib has low permeability.

Table 3 Permeability test results

(2) Study on the inhibition of cytochrome P450 isoenzymes

Experimental purpose: To determine the inhibitory effect of the test compound on the activity of human liver microsomal cytochrome P450 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4).

Experimental operation: First, the test compound (10mM) is gradient, and the working solution (100×final concentration) is prepared. The concentration of the working solution are: 5, 1.5, 0.5, 0.15, 0.05, 0.015 and 0.005mM. At the same time, prepare P450 co-workers Enzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) each positive inhibitor and its specific substrate mixture (5in 1) working solution; put the human liver microsomes frozen in the refrigerator at -80℃ on ice to thaw and wait for others The liver microsomes are completely dissolved and diluted with PB to prepare a working solution of a certain concentration (0.253mg/ml); add 20μL of the substrate mixture to the reaction plate (add 20μl PB to the Blank well) and 158μL of human liver microsomes The working solution is added to the reaction plate, and the reaction plate is placed on ice for use; at this time, 2μl of each concentration of the test compound (N=1) and specific inhibitor (N=2) are added to the corresponding wells, no inhibition Add the corresponding organic solvent to the agent (test compound or positive inhibitor) group as a control sample (test compound control sample is 1:1 DMSO: methanol, positive control sample is 1:9 DMSO: methanol,); in a 37°C water bath After pre-incubation for 10 minutes, add 20μl of Coenzyme Factor (NADPH) solution to the reaction plate and incubate in a 37°C water bath for 10 minutes; add 400μL of cold acetonitrile solution (internal standard: 200ng/mL Tolbutamide and Labetalol) to stop the reaction; Place the plate on a shaker and shake for 10 minutes; centrifuge at 4,000 rpm for 20 minutes; add 200 μL of supernatant to 100 μL of water to dilute the sample; finally, seal the plate, shake, shake, and perform LC/MS/MS detection.

Experimental results: Both the compound of the present invention and Sitravatinib have no obvious inhibitory effect on CYP1A2, and have a moderate inhibitory effect on CYP2C9 and CYP2C19. The compound of the present invention has weak and no inhibitory effects on CYP2D6 and CYP3A4 (substrate is midazolam), and Sitravatinib has strong inhibitory effects on CYP2D6 and CYP3A4 (substrate is midazolam).

Table 4 Experimental results of cytochrome P450 isoenzyme inhibition

(3) Study on plasma protein binding rate

Experimental purpose: to determine the protein binding rate of the test compound in the plasma of human, CD-1 mice and SD rats

Experimental operation: Take 796μL of blank plasma from human, CD-1 mice and SD rats, and add 4μL of test compound working solution (400μM) or warfarin working solution (400μM) to make the test compound in the plasma sample and warfarin The final Lin concentration is 2μM. Mix the sample thoroughly. The final concentration of DMSO in the organic phase is 0.5%; pipette 50μL of the test compound and warfarin plasma sample into the sample receiving plate (three parallel), immediately add the corresponding volume of the corresponding blank plasma or buffer to make each sample well The final volume is 100μL, the volume ratio of plasma:dialysis buffer is 1:1, and then 400μL of stop solution is added to these samples. This sample will be used as a T0 sample for recovery and stability determination. Store the T0 sample at 2-8°C and wait for subsequent processing with other dialyzed samples; add 150μL of test compound and warfarin plasma sample to the dosing end of each dialysis hole, and receive it at the corresponding dialysis hole Add 150μL of blank dialysis buffer to the end. Then, the dialysis plate was sealed with a gas-permeable membrane and placed in a humidified, 5% CO ₂ incubator, and incubated at 37° C. with shaking at about 100 rpm for 4-hr. After the dialysis, transfer 50 μL of the dialysis buffer sample and the dialysis plasma sample to a new sample receiving plate. Add a corresponding volume of blank plasma or buffer to the sample so that the final volume of each sample well is 100 μL, and the volume ratio of plasma:dialysis buffer is 1:1. All samples were analyzed by LC/MS/MS after protein precipitation, and the protein binding was calculated by the formula: %Unbound=100*FC/TC; %Bound=100-%Unbound; %Recovery=100*(FC+TC)/T0 Rate and recovery rate.

Experimental conclusions: Both the formate and Sitravatinib of compound 3 showed a higher plasma protein binding rate. The protein binding rate of compound 3 in the plasma of human, CD-1 mouse and SD rat were 99.7%, 99.7% and 99.8%, respectively. The protein binding rate of Sitravatinib in the plasma of human, CD-1 mouse and SD rat They are 99.95%, 99.97% and NA respectively (the value cannot be calculated because no free compound is detected at the free end).

(4) Studies on the metabolic stability of microsomes

Experimental purpose: to determine the stability of the test compound in liver microsomes of human, CD-1 mice and SD rats.

Experimental operation: First prepare 8 96-well plates, named T0, T5, T10, T20, T30, T60, NCF60 and BLANK; except for BLANK (add 10μL/well buffer), add 10μL/well of drug to each plate Solution; add the prepared microsomes to 7 plates (80μL/well), except for the T0 plate; add 10μL/well buffer to the NCF60 plate, incubate in a 37°C water bath, and start timing:

Then aliquot the prepared NADPH cofactor into a 96-well shallow-well plate as a sampling tank, and use a 96-channel pipette to divide 10 μL/well into a 37°C water bath to incubate to start the reaction:

Add 300 (μL/well) stop solution (cold acetonitrile, containing 100ng/mL methenine and 100ng/mL labetalol as internal standard) at each time point to stop the reaction, mix well; centrifuge at 4000 rpm for 20 minutes to precipitate Protein; After centrifugation, use a 96-channel pipette to transfer 100μL/well of the supernatant to a new plate with 300μL/well of HPLC water, mix well, and send it to the analyst for detection by LC/MS/MS.

Experimental results: The remaining percentages of the formate of compound 3 after 60 minutes of incubation in human, CD-1 mouse and SD rat liver microsomes are: 65.7%, 63.3% and 64.8%, respectively. Sitravatinib is in human and CD-1 The remaining percentages of mouse and SD rat liver microsomes incubated for 60 minutes were: 77.3%, 10.2%, and 34.4%, respectively. Compound 3 is metabolized at a moderate rate in the three species, Sitravatinib is metabolized at a moderate rate in human and SD rat liver microsomes, and it is metabolized faster in CD-1 mouse liver microsomes.

The evaluation results of in vitro DMPK properties are summarized in Table 5:

Table 5 Summary of evaluation results of in vitro DMPK properties

Experimental conclusion: The compound of the present invention is significantly better than the clinical phase III AXL&VEGFR2 inhibitor Sitravatinib in liver microsome stability (MMS), plasma protein binding rate (PPB), drug interaction (DDI) and in vitro permeability, which means This series of compounds have better metabolic stability; the improved permeability means that this series of compounds will be more easily absorbed, and at the same time, the risk of drug resistance due to active efflux is smaller; the CYP inhibitory activity of this series of compounds is low, and the drugs interact with each other. It has good properties of action, the safety risk of clinical combination with other drugs is smaller, and the developability is stronger.

Experimental Example 5: In vivo pharmacokinetic study

Experimental purpose: To study the pharmacokinetics of CD-1 mice after oral and intravenous injection of compound 3 or Sitravatinib

Experimental operation: Weigh appropriate amount of formate or Sitravatinib of compound 3, respectively dissolve them in a solvent of 5% DMSO/95% (20% HP-β-CD), stir and sonicate to prepare a clear solution of 0.4 mg/mL. Use the microporous membrane to filter for later use. CD-1 male mice aged 7-9 weeks were selected and administered intravenously with a volume of 5 mL/kg and a dose of 2 mg/kg.

Weigh appropriate amounts of compound 3 or Sitravatinib, respectively dissolve them in a solvent of 5% DMSO/95% (20% HP-β-CD), stir and sonicate to prepare a clear solution of 1 mg/mL. CD-1 male mice aged 7-9 weeks were selected for oral administration with a volume of 10 mL/kg and a dose of 10 mg/kg.

Collect whole blood for a certain period of time, prepare plasma, analyze drug concentration by LC-MS/MS method, and use Phoenix WinNonlin software (Pharsight, USA) to calculate pharmacokinetic parameters.

Experimental results:

After a single intravenous injection of 2 mg/kg Sitravatinib and compound 3 in male CD-1 mice, the plasma clearance (CL) was 7.21 and 6.18 mL/min/kg, and the steady-state apparent volume of distribution (Vd _ss ) was 2.24 And 0.72L/kg, the elimination half-life (T _1/2 ) is 3.87 and 1.58h, respectively, and the area under the plasma concentration curve (AUC _0-last ) from 0 o’clock to the last quantifiable time point is 7284 and 9645 nM·h, respectively .

After a single intragastric administration of 10 mg/kg Sitravatinib and compound 3 to male CD-1 mice, the bioavailability was 65.8% and 109%, and the AUC _0-last was 24003 and 53755 nM·h, respectively. The peak concentration (C _max ) Are 2530 and 10600 nM, and the peak time appears at 2.00 and 1.00 h after administration. The evaluation results of in vivo PK properties are summarized in Table 6.

Table 6 Summary of evaluation results of in vivo PK properties

Experimental results:

The compound of the present invention has good oral maximum plasma concentration C _max , half-life, clearance rate (CL), AUC and oral bioavailability.

Claims

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

in,

Ring A is selected from 5-membered heteroaryl groups, which contain 2 N heteroatoms;

Alternatively, ring A is selected from 6-membered heteroaryl groups, which contain up to 2 N atoms;

T 1 and T 2 are independently selected from C(R 5 ) and N;

R 1 and R 3 are each independently selected from H, F and CH 3 ;

R 2 chooses H, F and Cl;

R 4 is each independently selected from H, C 1-6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, and -LR 6 , the C 1-6 6 alkyl, C 1-6 heteroalkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl and -LR 6 are optionally substituted with 1, 2 or 3 R;

R 5 is selected from H and Cl;

R 6 is selected from H, OH, 5- to 6-membered heterocycloalkyl and C 1-3 alkyl;

L is selected from -CH 2 -, -CH 2 -CH 2 -and

n is selected from 0, 1, 2 and 3;

R is independently selected from F, Cl, Br, I, OH, CN, COOH, NH 2 , -NHCH 3 , -N(CH3) 2 , CH 3 , CH 2 CH 3 , CF 3 , -OCH 3 ,- OCH 2 CH 3 , -O-CH(CH 3 ) 2 , -C(=O)OCH 3 , -C(=O)CH 3 and -C(=O)CH 2 CH 3 ;

The 5-membered heteroaryl, 6-membered heteroaryl, C 1-6 heteroalkyl and 3-6 heterocycloalkyl each independently comprise 1, 2 or 3 independently selected from O, S, N and NH heteroatom or heteroatom group.
The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein ring A is selected from pyridyl, pyrazolyl and imidazolyl.
The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 4 is each independently selected from H, C 1-4 alkyl, C 1-4 heteroalkyl, 5--6 membered hetero Cycloalkyl and -LR 6 , the C 1-4 alkyl, C 1-4 heteroalkyl, 5-6 membered heterocycloalkyl and -LR 6 are optionally substituted with 1, 2 or 3 Rs.
The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 4 is each independently selected from H,

Said

Optionally substituted by 1, 2 or 3 R.
The compound or a pharmaceutically acceptable salt thereof according to claim 4, wherein R 4 is each independently selected from H,
The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the structural unit
Selected from
The compound or a pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein the structural unit
Selected from
The compound or a pharmaceutically acceptable salt thereof according to claim 7, wherein the structural unit
Selected from
The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein the compound is selected from

in,

R 41 is selected from H, C 1-4 alkyl, C 1-4 heteroalkyl, 6-membered heterocycloalkyl and -LR 6 , the H, C 1-4 alkyl, C 1-4 heteroalkyl , 6-membered heterocycloalkyl and -LR 6 are optionally substituted with 1, 2 or 3 R;

R 42 is selected from H and -LR 6 , said -LR 6 is optionally substituted by 1, 2 or 3 R;

R 6 is selected from H, OH, 6-membered heterocycloalkyl and C 1-3 alkyl;

R 2 , L and R are as defined in claim 1.
The compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein the compound is selected from

R 1 , R 2 , R 3 and R 4 are as defined in claims 1-5.
The compound or a pharmaceutically acceptable salt thereof, which is selected from
A pharmaceutical composition comprising a therapeutically effective amount of the compound or pharmaceutically acceptable salt according to any one of claims 1 to 11 as an active ingredient and a pharmaceutically acceptable carrier.
Use of the compound or pharmaceutically acceptable salt according to any one of claims 1 to 11 or the composition according to claim 12 in the preparation of Axl and/or VEGFR-2 inhibitors.
The application according to claim 13, wherein the Axl and/or VEGFR-2 inhibitor is a medicine for treating cancer.