Compounds containing phenylsulfonamide

Abstract

The present invention discloses a class of compounds containing aromatic sulfonamides, and specifically discloses a compound represented by formula (II), a pharmaceutically acceptable salt thereof or an isomer thereof.

Description

[0001] This application claims the priority of Chinese patent application CN202010363156.1 filed on April 30, 2020 and Chinese patent application CN202110145140.8 filed on February 2, 2021. This application cites the full text of the above Chinese patent application. Technical Field

[0002] The present invention relates to a class of compounds containing phenylsulfonamide, and specifically to a compound represented by formula (II), a pharmaceutically acceptable salt thereof or an isomer thereof. Background Art

[0003] The mitogen-activated protein kinase (MAPK) pathway exists in a series of cellular processes such as cell proliferation, differentiation, apoptosis and stress response. Among them, the RAS-RAF-MEK-ERK pathway is one of the most well-known MAPK pathways. This pathway first starts with the binding of extracellular growth factors (PDGF or EGF) to transmembrane receptors (EGFR or PDGFR, etc.), activating the receptors. The activated receptors bind to GTP and activate the RAS protein in the membrane through the guanine exchange factor (SOS); the activated RAS further indirectly phosphorylates RAF; the activated RAF is phosphorylated on two serine residues of MEK1 / 2; the activated MEK1 / 2 in turn activates its downstream substrate ERK1 / 2; the phosphorylated ERK1 / 2 dimerizes and moves to the nucleus and accumulates. ERK in the nucleus is involved in many cellular functions, including nuclear transport, signal transduction, DNA repair, and mRNA processing and translation. If the genes involved in the pathway mutate or growth factors, downstream signaling proteins or protein kinases are overexpressed, the cell pathway will be continuously activated, cell proliferation will be uncontrolled and eventually lead to tumor formation. For example, about 30% of human cancer cells have RAS mutations, of which KRAS mutations are the most common subtype of RAS mutations. KRAS-mutated tumors account for about 22% of all human tumor cells, of which 70-90% of pancreatic cancers, 10-20% of non-small cell lung cancers and 25-35% of colorectal cancers have KRAS mutations; about 8% of tumors have BRAF mutations, of which 50-60% of melanomas and 40-60% of papillary thyroid cancers have BRAF mutations.

[0004] Extracellular signal-regulated kinase (ERK 1 / 2) is an important member of the MAPK family. As the "final manager" downstream of the RAS / RAF / MEK / ERK pathway, targeted inhibition of ERK1 / 2 is expected to be used to treat carcinogenesis caused by abnormal activation of the MAPK pathway (activation mutations such as RAS / RAF / MEK), and may also be effective for patients who develop resistance to RAF or MEK inhibitors due to reactivation of ERK1 / 2. According to many preclinical reports, MAPK pathway inhibitors can effectively inhibit cancer cells with BRAF and RAS mutations. For example, BRAF inhibitors vemurafenib and dabrafenib and MEK inhibitor trametinib have been approved for the treatment of BRAF-mutated melanoma. However, these drugs still have resistance issues. The mechanisms of BRAF inhibitor resistance have been confirmed, including MEK transactivation of CRAF, RTK, upregulation of NRAS signaling, and MEK activating mutations; MEK inhibitor resistance mechanisms include MEK mutations that reduce its binding to drugs or enhance the activity of MEK itself, as well as BRAF or KRAS amplification. Whether it is resistance to RAF inhibitors or MEK inhibitors, the RAS-RAF-MEK-ERK pathway will be reactivated, leading to the continued proliferation of cancer cells. Therefore, the development of a new type of dual-mechanism ERK inhibitor will not only be effective for patients with mutations in the MAPK signaling pathway, but also for patients who are resistant to BRAF and MEK inhibitors. Summary of the invention

[0005] The present invention provides a compound represented by formula (II) or a pharmaceutically acceptable salt thereof,

[0006]

[0007] in,

[0008] T1 is CH or N;

[0009] n is 1 or 2;

[0010] R1 and R2 are each independently H, D, F, Cl or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br and I;

[0011] or R1 and R2 together with the carbon atom to which they are attached form R3 and R4 are each independently H or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH;

[0012] R5 and R6 are each independently H or C 1-3Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3;

[0013] R7 is phenyl or pyridyl, wherein the phenyl and pyridyl groups are optionally substituted by 1, 2, 3 or 4 R a replaced by;

[0014] R8 is H, F, Cl or Br;

[0015] R9 is tetrahydro-2H-pyranyl, wherein the tetrahydro-2H-pyranyl is optionally substituted with 1, 2, 3 or 4 R b replaced by;

[0016] Each R a are independently F, Cl, Br, I, C 1-3 Alkyl, C 1-3 Alkoxy, NH-C 1-3 Alkyl or N-(C 1-3 alkyl)2, wherein the C 1-3 Alkyl, C 1-3 Alkoxy, -NH-C 1-3 Alkyl and -N-(C 1-3 alkyl)2 are each independently optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH;

[0017] Each R b are independently F, Cl, Br, I, D or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH.

[0018] The present invention also provides a compound represented by formula (II) or a pharmaceutically acceptable salt thereof,

[0019]

[0020] in,

[0021] T1 is CH or N;

[0022] n is 1 or 2;

[0023] R1 and R2 are each independently H, F, Cl or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br and I;

[0024] or R1 and R2 together with the carbon atom to which they are attached form R3 and R4 are each independently H or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH;

[0025] R5 and R6 are each independently H or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3;

[0026] R7 is phenyl or pyridyl, wherein the phenyl and pyridyl groups are optionally substituted by 1, 2, 3 or 4 R a replaced by;

[0027] R8 is H, F, Cl or Br;

[0028] R9 is tetrahydro-2H-pyranyl, wherein the tetrahydro-2H-pyranyl is optionally substituted with 1, 2, 3 or 4 R b replaced by;

[0029] Each R a are independently F, Cl, Br, I, C 1-3 Alkyl, C 1-3 Alkoxy, NH-C 1-3 Alkyl or N-(C 1-3 alkyl)2, wherein the C 1-3 Alkyl, C 1-3 Alkoxy, -NH-C 1-3 Alkyl and -N-(C 1-3 alkyl)2 are each independently optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH;

[0030] Each R b are independently F, Cl, Br, I, D or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH.

[0031] The present invention also provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

[0032]

[0033] Where n is 1 or 2;

[0034] R1 and R2 are each independently H, F, Cl or C 1-3 Alkyl, wherein the C 1-3The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br and I;

[0035] or R1 and R2 together with the carbon atom to which they are attached form

[0036] R3 and R4 are each independently H or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br and I;

[0037] R5 and R6 are each independently H or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3;

[0038] R7 is phenyl, wherein the phenyl group is optionally substituted by 1, 2, 3 or 4 R a replaced by;

[0039] R8 is H, F or Cl;

[0040] R9 is tetrahydro-2H-pyranyl, wherein the tetrahydro-2H-pyranyl is optionally substituted with 1, 2, 3 or 4 R b replaced by;

[0041] Each R a are independently F, Cl, Br, I, C 1-3 Alkyl or C 1-3 Alkoxy, wherein the C 1-3 Alkyl and C 1-3 The alkoxy group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH;

[0042] Each R b are independently F, Cl, Br, I or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH.

[0043] In some embodiments of the present invention, the above compound has a structure shown in formula (I-1) or (I-2):

[0044]

[0045] wherein R1, R2, R3, R4, R5, R6, R7, R8 and R9 are as defined in the present invention.

[0046] In some embodiments of the present invention, each of the above R bindependently represents F, Cl, Br, I, D or -CH3, and the other variables are as defined herein.

[0047] In some embodiments of the present invention, each of the above R b independently F, Cl, Br, I or -CH3, and the other variables are as defined herein.

[0048] In some embodiments of the present invention, the above R9 is It is stated Optional 1, 2, 3 or 4 R b Replaced by R b and other variables are as defined in the present invention.

[0049] In some embodiments of the present invention, the above R9 is Other variables are as defined in the present invention.

[0050] In some embodiments of the present invention, the above R9 is R b and other variables are as defined in the present invention.

[0051] In some embodiments of the present invention, the above-mentioned compound has a structure shown in formula (III-1) or (III-2):

[0052]

[0053] in,

[0054] m is 0, 1, 2, 3 or 4;

[0055] T1, R b , R1, R2, R3, R4, R5, R6, R7 and R8 are as defined in the present invention.

[0056] In some embodiments of the present invention, the above compound has a structure shown in formula (I-3) or (I-4):

[0057]

[0058] wherein R1, R2, R3, R4, R5, R6, R7 and R8 are as defined in the present invention.

[0059] In some embodiments of the present invention, the above R1 and R2 are each independently H, D, F, Cl or -CH3, and other variables are as defined in the present invention.

[0060] In some embodiments of the present invention, the above R1 and R2 are each independently H, F, Cl or -CH3, and other variables are as defined in the present invention.

[0061] In some embodiments of the present invention, the above R1 and R2 are each independently H, and other variables are as defined in the present invention.

[0062] In some embodiments of the present invention, the above R1 and R2 together with the carbon atom to which they are attached form Other variables are as defined herein.

[0063] In some embodiments of the present invention, the above R3 and R4 are each independently H or -CH3, wherein the -CH3 is optionally substituted by 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH, and other variables are as defined in the present invention.

[0064] In some embodiments of the present invention, the above R3 and R4 are each independently H or -CH3, wherein the -CH3 is optionally substituted by 1, 2 or 3 substituents independently selected from F, Cl, Br and I, and other variables are as defined in the present invention.

[0065] In some embodiments of the present invention, the above R3 and R4 are each independently H or -CH3, and other variables are as defined in the present invention.

[0066] In some embodiments of the present invention, the above R5 and R6 are each independently H or -CH3, wherein the -CH3 is optionally substituted by 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3, and other variables are as defined in the present invention.

[0067] In some embodiments of the present invention, the above R5 and R6 are each independently H, -CH3 or Other variables are as defined in the present invention.

[0068] In some embodiments of the present invention, R5 and R6 are each independently H or Other variables are as defined herein.

[0069] In some embodiments of the present invention, the above compound has a structure shown in formula (III-3) or (III-4):

[0070]

[0071] in,

[0072] m is 0, 1, 2, 3 or 4;

[0073] T1, R b , R7 and R8 are as defined in the present invention;

[0074] R3 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH;

[0075] R5 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3.

[0076] In some embodiments of the present invention, the above compound has a structure shown in formula (I-5) or (I-6):

[0077]

[0078] Wherein, R7 and R8 are as defined in the present invention;

[0079] R3 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH;

[0080] R5 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3.

[0081] In some embodiments of the present invention, the above compound has a structure shown in formula (I-5) or (I-6):

[0082]

[0083] Wherein, R7 and R8 are as defined in the present invention;

[0084] R3 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br and I;

[0085] R5 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3.

[0086] In some embodiments of the present invention, each of the above R a are independently F, Cl, Br, I, -CH3, -OCH3, -NH-CH3 or Other variables are as defined herein.

[0087] In some embodiments of the present invention, each of the above R a independently represents F, Cl, Br, I, -CH3 or -OCH3, and other variables are as defined herein.

[0088] In some embodiments of the present invention, each of the above R a are independently F, Cl, -CH3, -OCH3 or Other variables are as defined herein.

[0089] In some embodiments of the present invention, each of the above R a independently F or -OCH3, and the other variables are as defined herein.

[0090] In some embodiments of the present invention, the above R7 is R a and other variables are as defined in the present invention.

[0091] In some embodiments of the present invention, the above R7 is R a and other variables are as defined in the present invention.

[0092] In some embodiments of the present invention, the above R7 is Other variables are as defined in the present invention.

[0093] In some embodiments of the present invention, the above R7 is Other variables are as defined in the present invention.

[0094] In some embodiments of the present invention, the above R7 is Other variables are as defined in the present invention.

[0095] In some embodiments of the present invention, the above R7 is Other variables are as defined in the present invention. In the present invention, there are also some schemes which are obtained by any combination of the above variables.

[0096] The present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof:

[0097]

[0098]

[0099] The present invention provides a compound of the following formula or a pharmaceutically acceptable salt thereof:

[0100]

[0101]

[0102] The present invention also provides the use of the above compound and pharmaceutically acceptable salts thereof in the preparation of ERK1 / 2 inhibitor drugs.

[0103] Technical Effects

[0104] The compound of the present invention has good inhibitory activity on ERK1 / 2. It is expected to be used for carcinogenesis caused by abnormal activation of MAPK signaling pathway (RAS / RAF / MEK and other activation mutations), and may also be effective for patients who develop resistance to RAF or MEK inhibitors due to reactivation of ERK1 / 2; the compound of the present invention has good oral absorption in mice and dogs, low clearance rate, high exposure, and good bioavailability; the compound of the present invention has a significant inhibitory effect on the growth of tumor-bearing mice in a subcutaneous xenograft tumor model of human lung cancer Calu-6 cells.

[0105] Definition and Description

[0106] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

[0107] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0108] The term "pharmaceutically acceptable salt" refers to salts of compounds of the invention, prepared from compounds of the invention having specific substituents with relatively nontoxic acids or bases. When the compounds of the invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When the compounds of the invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid and methanesulfonic acid, and 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, and thus can be converted into any base or acid addition salt.

[0109] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0110] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention.

[0111] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0112] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability of a ring carbon atom to rotate freely about double bonds or single bonds forming a ring.

[0113] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0114] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0115] Unless otherwise specified, the key is a solid wedge. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond. and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Indicates a wedge-shaped solid key or dotted wedge key Or use a wavy line Represents a straight solid bond and straight dashed key

[0116] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0117] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0118] Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (for example, a carbamate is generated from an amine). The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more atoms that constitute the compound. For example, the compound can be labeled with a radioactive isotope, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For another example, deuterium can be substituted for hydrogen to form a deuterated drug. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing therapeutic effects, and extending the biological half-life of drugs. All isotopic composition changes of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention. "Optional" or "optionally" means that the event or situation described later may but does not necessarily occur, and the description includes the case where the event or situation occurs and the case where the event or situation does not occur.

[0119] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include a variant of deuterium and hydrogen, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or not substituted, and unless otherwise specified, the type and number of the substituents may be arbitrary on the basis of chemical achievable.

[0120] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice. In addition, combinations of substituents and / or variants thereof are permitted only if such combinations result in stable compounds.

[0121] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0122] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0123] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A. When the listed substituent does not specify which atom it is connected to the substituted group through, the substituent can be bonded through any atom of it. For example, pyridyl as a substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

[0124] When the listed linking group does not indicate its linking direction, its linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of the reading order from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0125] 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. The chemical bonds connecting the sites to other groups can be straight solid bonds. Straight dotted key or wavy line For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy lines in the phenyl group represent the connection to other groups through the carbon atoms at positions 1 and 2 in the phenyl group.

[0126] Unless otherwise specified, the number of atoms in a ring is generally defined as the ring member number, for example, a "5-7 membered ring" refers to a "ring" having 5-7 atoms arranged around it.

[0127] Unless otherwise specified, "5-membered ring" means a cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl or heteroaryl consisting of 5 ring atoms. The ring includes a monocyclic ring, and also includes a bicyclic ring system such as a spirocyclic ring, a cyclic ring and a bridged ring. Unless otherwise specified, the ring optionally contains 1, 2 or 3 heteroatoms independently selected from O, S and N. The term "ring" also includes a ring system containing at least one ring, each of which "ring" independently meets the above definition.

[0128] Unless otherwise specified, the term “C 1-3 "Alkyl" is used to refer to a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl, etc.; it may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

[0129] Unless otherwise specified, the term “C 1-3 "Haloalkyl" refers to monohaloalkyl and polyhaloalkyl groups containing 1 to 3 carbon atoms. 1-3 Haloalkyl includes C 1-2 , C 2-3 , C3, C2 and C1 haloalkyl, etc. 1-3 Examples of haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, pentafluoroethyl, pentachloroethyl, 3-bromopropyl, and the like.

[0130] Unless otherwise specified, the term “C 1-3 "Alkoxy" refers to those alkyl groups containing 1 to 3 carbon atoms which are attached to the rest of the molecule via an oxygen atom. 1-3 Alkoxy includes C 1-2 , C 2-3, C3 and C2 alkoxy, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), and the like.

[0131] Unless otherwise specified, the term “C 6-10 Aromatic rings" and "C 6-10 Aryl" can be used interchangeably, the term "C 6-10 Aromatic ring" or "C 6-10 "Aryl" means a cyclic hydrocarbon group composed of 6 to 10 carbon atoms with a conjugated π electron system, which can be a monocyclic, fused bicyclic or fused tricyclic system, in which each ring is aromatic. It can be monovalent, divalent or polyvalent, C 6-10 Aryl includes C 6-9 , C9, C 10 and C6 aryl, etc. 6-10 Examples of aryl groups include, but are not limited to, phenyl, naphthyl (including 1-naphthyl and 2-naphthyl, etc.).

[0132] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 Etc.; similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, 3-12-membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring, 10-membered ring, 11-membered ring, and 12-membered ring, and also includes any range from n to n+m, for example, 3-12-membered ring includes 3-6-membered ring, 3-9-membered ring, 5-6-membered ring, 5-7-membered ring, 6-7-membered ring, 6-8-membered ring, and 6-10-membered ring, etc.

[0133] The term "D" refers to deuterium, an isotope of hydrogen, also known by the chemical symbol 2 H, also known as heavy hydrogen, is composed of one proton, one neutron and one electron.

[0134] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., an affinity substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chlorine, bromine, iodine; sulfonate groups, such as mesylate, tosylate, p-brosylate, p-toluenesulfonate, etc.; acyloxy groups, such as acetoxy, trifluoroacetoxy, etc.

[0135] The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group" or "thiol protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions of the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and tert-butyl; acyl groups, for example, alkanoyl groups (such as acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like.

[0136] 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, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention.

[0137] The structure of the compound of the present invention can be confirmed by conventional methods known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data of the cultured single crystal using a Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is: / ω scanning, after collecting relevant data, the direct method (Shelxs97) is further used to analyze the crystal structure to confirm the absolute configuration.

[0138] The solvent used in the present invention is commercially available.

[0139] The present invention uses the following abbreviations: BF3·Et2O represents boron trifluoride ether adduct; DMSO represents dimethyl sulfoxide; DMF represents N,N-dimethylformamide; DPBS represents Dulbecco's phosphate buffer; EDCI represents 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide; HOBt represents 1-hydroxybenzotriazole; HPLC represents high pressure liquid chromatography; LCMS represents liquid chromatography-mass spectrometry; MeOH represents methanol; NMM represents nitrogenmethylmorpholine; Pd(dppf)Cl2·CH2Cl2 represents [1,1'-bis( [(1,2-diphenylphosphino)ferrocene]palladium dichloride dichloromethane adduct; Pd(PPh3)4 represents tetrakistriphenylphosphine palladium; PBS represents phosphate buffered saline; HATU represents O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; PMB represents p-methoxybenzyl; NMP represents nitrogenmethylpyrrolidone; DIEA represents N,N-diisopropylethylamine; LiAlD4 represents lithium aluminum tetrahydride deuterate; MsCl represents methanesulfonyl chloride; BNS represents N-bromosuccinimide; AIBN represents azobisisobutyronitrile. BRIEF DESCRIPTION OF THE DRAWINGS

[0140] Figure 1 : Tumor volume of each group at different time points;

[0141] Figure 2 : Effects of the test substances on the body weight of mice. DETAILED DESCRIPTION

[0142] The present invention is described in detail below by way of examples, but it is not intended to impose any adverse limitations on the present invention. The present invention has been described in detail herein, and specific embodiments thereof are also disclosed therein. It will be apparent to those skilled in the art that various changes and modifications may be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0143] Example 1

[0144]

[0145] Compound 1A:

[0146]

[0147] Compound SM1 (4 g, 17.53 mmol) and palladium / carbon (400 mg, 17.53 mmol, 10% purity) were dissolved in methanol (40 ml), and the reaction solution was replaced with hydrogen three times to expel air, and then stirred at 15° C. for 2 hours under the protection of hydrogen (15 psi). The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain crude product 1A, which was directly used in the next reaction. 1H NMR (400MHz, DMSO-d6) δ = 7.59 (d, J = 8.4Hz, 1H), 6.94 (d, J = 2.0Hz, 1H), 6.89 (dd, J = 1.9, 8.5Hz, 1H).

[0148] Compound 1B:

[0149]

[0150] At 0°C, a solution of compound 1A (2.66 g, 13.42 mmol) in acetonitrile (30 ml) was added dropwise to a solution of copper bromide (3.00 g, 13.43 mmol, 628.93 μl) and tert-butyl nitrite (3.38 g, 32.79 mmol, 3.9 ml) in acetonitrile (30 ml). After the addition, the reaction solution was stirred at 0°C for 1 hour. Then, it was stirred at 15°C for 15 hours. The reaction solution was diluted with water (100 ml), and hydrochloric acid solution (2 mol) was added to adjust the pH to below 2. Then, it was extracted with ethyl acetate (100 ml x 2 times). The organic phases were combined and concentrated under reduced pressure to obtain compound 1B. 1 H NMR (400MHz, DMSO-d6) δ = 8.51 (d, J = 1.2 Hz, 1H), 8.09 ( dd, J = 1.7, 8.2 Hz, 1H), 7.87 ( d, J = 8.2 Hz, 1H).

[0151] Compound 1C:

[0152]

[0153] Compound 1B (4.26 g, 16.25 mmol) was dissolved in tetrahydrofuran (50 ml), and sodium borohydride (6.40 g, 169.05 mmol) was added. The reaction solution was cooled to -8°C, and then BF3·Et2O (26.45 g, 186.36 mmol, 23 ml) was added dropwise at -8°C. The mixture was stirred at -8°C for 10 minutes. Then the mixture was stirred at 80°C for 2 hours. The reaction solution was poured into ice water (120 ml), and the pH of the reaction solution was adjusted to 10 with a 2 molar sodium hydroxide (100 ml) solution. The reaction solution was extracted with ethyl acetate (100 ml). The organic phase was extracted with a 2 molar sodium hydroxide (50 ml × 3 times) aqueous solution. The combined aqueous phase was adjusted to pH 2 with a 2 molar hydrochloric acid (500 ml) solution, and then extracted with ethyl acetate (300 ml × 2 times), and the combined organic phases were concentrated under reduced pressure to obtain a crude compound 1C. 1H NMR (400MHz, DMSO-d6) δ = 8.12 (d, J = 1.6 Hz, 1H), 7.97 ( br s, 1H), 7.87 ( dd, J = 1.8, 8.2 Hz, 1H), 7.53 ( d, J = 8.3 Hz, 1H), 4.37 ( br s, 2H).

[0154] Compound 1D:

[0155]

[0156] Under nitrogen protection, compound 1C (3.1 g, 12.50 mmol), ethyl 2-bromo-(2S) propionate (3.40 g, 18.78 mmol) and potassium carbonate (5.06 g, 36.65 mmol) were dissolved in DMF (30 ml), and the mixture was stirred at 20°C for 1 hour. The reaction solution was diluted with 50 ml of water and then extracted with ethyl acetate (50 ml x 3 times). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-5:1) to obtain compound 1D. 1 H NMR (400MHz, CDCl3) δ = 7.93 (d, J = 1.5Hz, 1H), 7.72 (dd, J = 1.8, 8.2Hz, 1H), 7.30 (d, J = 8.3Hz, 1H), 4.82 (d,J=14.1Hz,1H),4.59-4.46(m,2H),4.20(dq,J=1.4,7.2Hz,2H),1.63(d,J=7.3Hz,3H),1.26(s,3H).

[0157] Compound 1E:

[0158]

[0159] Under nitrogen protection, bispinacol borate (1.19 g, 4.69 mmol), compound 1D (1.36 g, 3.91 mmol), Pd(dppf)Cl2·CH2Cl2 (319 mg, 390.63 μmol) and potassium acetate (767 mg, 7.82 mmol) were dissolved in dioxane (14 ml), and the mixture was stirred at 90°C for 16 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude compound 1E. Compound 1F:

[0160]

[0161] Under nitrogen protection, compound 1E (2.53 g, 6.40 mmol), 2,4,5-trichloropyrimidine (2.35 g, 12.81 mmol), Pd(PPh3)4 (740 mg, 640.38 μmol) and sodium carbonate (1.36 g, 12.83 mmol) were dissolved in dioxane (24 ml) and water (8 ml), and the mixture was stirred at 90°C for 2 hours. The reaction solution was filtered, ethyl acetate (50 ml) was added to the filtrate, and washed with saturated sodium chloride solution (50 ml × 3 times). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-4:1) to obtain compound 1F. 1 H NMR (400MHz, CDCl3) δ = 8.72 (s, 1H), 8.42 (d, J = 1.1Hz, 1H), 8.19 (dd, J = 1.7, 8.1Hz, 1H), 7.59 (d, J = 8.1Hz, 1H), 4 .99(d,J=14.5Hz,1H),4.71-4.54(m,2H),4.22(dq,J=1.5,7.2Hz,2H),1.67(d,J=7.2Hz,3H),1.30-1.26(m,3H).

[0162] Compound 1G:

[0163]

[0164] Under nitrogen protection, compound 1F (865 mg, 2.08 mmol), tetrahydropyran-4-amine (420 mg, 4.15 mmol), DIEA (675.22 mg, 5.22 mmol, 910 μL) were dissolved in dioxane (9 mL), and the mixture was stirred at 90°C for 16 hours. The reaction solution was diluted with ethyl acetate (20 mL) and washed with 1M hydrochloric acid (20 mL × 1 time) aqueous solution, the aqueous phase was extracted with ethyl acetate (20 mL × 2 times), the combined organic phase was washed with brine (20 mL × 1 time), dried, and concentrated under reduced pressure to obtain crude compound 1G. 1H NMR (400MHz, CDCl3) δ = 8.33 (s, 1H), 8.27 (s, 1H), 8.08 (dd, J = 1.3, 8.1Hz, 1H), 7.52 (d, J = 8.1Hz, 1H), 5.34 (br s,1H),4.95(d,J=14.3Hz,1H),4.64-4.58(m,2H),4.22(dq,J=1.4,7.1Hz,2H),4.03-3.98(m,2H),3.56( dt,J=1.7,11.5Hz,2H),2.07-2.02(m,2H),1.67(d,J=7.4Hz,3H),1.61-1.55(m,2H),1.31-1.27(m,3H).

[0165] Compound 1H:

[0166]

[0167] To a solution of compound 1G (1.03 g, 2.14 mmol) in tetrahydrofuran (10 ml) and methanol (10 ml) was added lithium hydroxide monohydrate (4 mol, 1.1 ml). The mixture was stirred at 20°C for 16 hours. The reaction solution was concentrated under reduced pressure, and the pH was adjusted to 4 with 2 molar hydrochloric acid (2 ml) solution, and then extracted with ethyl acetate (20 ml x 3 times). The combined organic phase was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude compound 1H. 1 H NMR (400MHz, CDCl3) δ = 8.32 (br s, 1H), 8.23 ​​(s, 1H), 8.03 (br d, J = 7.5Hz, 1H), 7.52 (d, J = 8.0Hz, 1H), 6.15-6.07 (m, 1H), 4.83 (br d,J=13.7Hz,1H),4.67-4.55(m,3H),4.02-3.96(m,3H),3.58-3.51(m,2H),2.02(br d, J=10.7Hz, 2H), 1.69 (d, J=7.3Hz, 3H), 1.27 (t, J=7.2Hz, 2H).

[0168] Compound 1:

[0169]

[0170] Under nitrogen protection, compound 1H (247 mg, 545.36 μmol), (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (101 mg, 545.37 μmol), HOBt (82 mg, 606.85 μmol), NMM (61 mg, 603.08 μmol, 66.30 μl) and EDCI (115 mg, 599.89 μmol) were dissolved in a mixed solvent of DMF (3 ml) and dichloromethane (6 ml), and the mixture was stirred at 20°C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain a residue, which was diluted with dichloromethane (40 ml) and then washed with aqueous hydrochloric acid solution (2 mol, 40 ml × 2 times). The organic phases were combined and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (trifluoroacetic acid system) to obtain the free base of compound 1. 1 H NMR (400MHz, MeOH-d4) δ = 8.38 (s, 1H), 8.24 (d, J = 0.9Hz, 1H), 8.16 (dd, J = 1.5, 8.1Hz, 1H), 7.71 (d, J = 8.1Hz, 1H), 6.77 (s, 1H), 6.70 (br d,J=9.4Hz,1H),6.65-6.55(m,1H),4.99-4.91(m,2H),4.71(d,J=14.7Hz,1H),4.64-4.59(m, 1H),4.09-3.95(m,3H),3.81(s,3H),3.79-3.67(m,2H),3.55(dt,J=1.9,11.6Hz,2H),2.01(br dd,J=2.0,12.5Hz,2H),1.70-1.57(m,5H). LCMS(ESI)m / z:620.3[M+1].

[0171] Example 2

[0172]

[0173] Compound 2B:

[0174]

[0175] Compound 2A (74.76 mg, 480.45 μmol, HCl) and DIEA (186.28 mg, 1.44 mmol 251.06 μl) were added to a solution of compound 1F (200 mg, 480.45 μmol) in dioxane (3 ml), and the reaction solution was stirred at 90° C. for 24 hours. After the reaction, the reaction solution was diluted with ethyl acetate (20 ml) and washed with a 1 molar hydrochloric acid aqueous solution (20 ml × 1 time), the aqueous phase was extracted with ethyl acetate (20 ml × 2 times), and the combined organic phase was washed with brine (20 ml × 1 time), dried, and concentrated under reduced pressure to obtain a crude compound 2B. 1 H NMR (400MHz, CDCl3) δ = 8.37 (s, 1H), 8.29 (s, 1H), 8.09 (dd, 1H, J = 1.5, 8.2Hz), 7.53 (d, 1H, J = 8.1Hz), 5.32 (br d,1H,J=8.1Hz),4.96(d,1H,J=14.3Hz),4.6-4.7(m,2H),4.46(s,2H),4.1-4.2(m,2H),3.8-3.9(m,1H) ,3.5-3.6(m,2H),2.2-2.4(m,1H),1.67(d,3H,J=7.3Hz),1.3-1.3(m,3H).LCMS(ESI):m / z:416.2[M+1].

[0176] Compound 2C:

[0177]

[0178] To a solution of compound 2B (60 mg, 120.25 μmol) in tetrahydrofuran (3 ml) was added water and lithium hydroxide (15.14 mg, 360.75 μmol) in water (0.5 ml), and the reaction solution was stirred at 25°C for 1 hour. After the reaction was complete, the reaction solution was concentrated under reduced pressure, adjusted to pH = 4 with 2 molar hydrochloric acid (2 ml) solution, and then extracted with ethyl acetate (20 ml × 3 times). The combined organic phase was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude compound 2C. LCMS (ESI): m / z: 471.0 [M+1].

[0179] Compound 2C:

[0180]

[0181] To a solution of compound 2C (50 mg, 106.18 μmol) in dichloromethane (3 ml) were added (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (28.24 mg, 127.42 μmol) and DIEA (41.17 mg, 318.54 μmol, 55.48 μl), the mixture was stirred at 0°C for 15 minutes, and then HATU (48.45 mg, 127.42 μmol) was added to the mixture, and the reaction solution was stirred at 0°C for 1 hour. The reaction solution was adjusted to pH <7 with a hydrochloric acid (2M) aqueous solution, then extracted with dichloromethane (10 ml × 2 times), and then washed with water (10 × 1 time). The organic phases were combined and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 2. 1 H NMR (CD3OD, 400MHz) δ = 8.41 (s, 1H), 8.26 (d, 1H, J = 1.1Hz), 8.18 (dd, 1H, J = 1.6, 8.1Hz), 7.72 (d, 1H, J = 8.1 Hz),6.77(s,1H),6.70(td,1H,J=1.7,9.3Hz),6.6-6.6(m,1H),5.0-5.0(m,1H),4.72(d,1H,J=14.8Hz),4 .5-4.7(m,4H),4.2-4.4(m,1H),4.07(dt,1H,J=3.9,11.2Hz),3.9-4.0(m,1H),3.81(s,3H),3.7-3.8(m,2 H),3.5-3.6(m,2H),2.1-2.3(m,1H),1.7-1.8(m,1H),1.62(d,3H,J=7.1Hz).LCMS(ESI):m / z:638.0[M+1].

[0182] Example 3

[0183]

[0184]

[0185] Compound 3B:

[0186]

[0187] Under nitrogen protection, compound 1E (500 mg, 1.26 mmol), 3A (454 mg, 1.99 mmol, 255.06 μL), Pd(PPh3)4 (145 mg, 125.48 μM) and sodium carbonate (409 mg, 3.86 mmol) were dissolved in dioxane (8 ml) and water (2 ml), and the mixture was stirred at 100°C for 1 hour. The reaction solution was filtered, ethyl acetate (30 ml) was added to the filtrate, and washed with saturated sodium chloride solution (30 ml × 1 time). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound 3B. 1 H NMR (400MHz, CDCl3) δ = 8.84 (s, 1H), 8.35 (d, J = 1.3Hz, 1H), 8.12 (dd, J = 1.6, 8.1Hz, 1H), 7.58 (d, J = 8.1Hz, 1H), 4.98 (d, J = 1 4.4Hz,1H),4.67-4.57(m,2H),4.25-4.18(m,2H),1.67(d,J=7.3Hz,3H),1.31-1.27(m,3H).LCMS(ESI):m / z:527.2[M+1]. LCMS(ESI):m / z:461.9[M+1].

[0188] Compound 3C:

[0189]

[0190] DIEA (61 mg, 471.98 μmol, 82.21 μL) and tetrahydropyran-4-amine (66 mg, 652.52 μmol) were added to a solution of compound 3B (100 mg, 217.05 μmol) in dioxane (3 ml). The reaction solution was stirred at 90°C for 16 hours. After the reaction was completed, the mixture was diluted with water (10 ml) and extracted with ethyl acetate (10 ml x 2 times). The combined organic phases were concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1 elution) to obtain compound 3C. LCMS (ESI): m / z: 527.2 [M+1].

[0191] Compound 3D:

[0192]

[0193] To a solution of compound 3C (100 mg, 190.33 μmol) in tetrahydrofuran (1 ml) and ethanol (1 ml) was added water and a solution of lithium hydroxide (12 mg, 285.96 μmol) in water (1 ml), and the reaction solution was stirred at 25°C for 1 hour. After the reaction was complete, the reaction solution was concentrated under reduced pressure, the reaction solution was adjusted to pH = 2 with an aqueous solution of hydrochloric acid (2 mol), and then extracted with ethyl acetate (10 ml × 3 times). The combined organic phase was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude compound 3D. LCMS (ESI): m / z: 497.2 [M+1].

[0194] Compound 3:

[0195]

[0196] To a solution of compound 3D (70 mg, 140.74 μmol) in dichloromethane (3 ml) were added (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (58 mg, 261.67 μmol) and DIEA (74.20 mg, 574.11 μmol, 100 μl), and the reaction solution was stirred at 0°C for 5 minutes. HATU (70 mg, 184.10 μmol) was then added to the reaction solution and the reaction was continued for 2 hours. The reaction solution was adjusted to pH <7 with a hydrochloric acid (2M) aqueous solution, then extracted with dichloromethane (10 ml × 2 times), and then washed with water (10 ml × 1 time). The combined organic phases were filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 3. 1 H NMR (400MHz, CD3OD) δ = 8.46 (s, 1H), 8.15 (d, J = 1.1Hz, 1H), 8.08 (dd, J = 1.6, 8.1Hz, 1H), 7.68 (d ,J=8.1Hz,1H),6.75(s,1H),6.72-6.65(m,1H),6.59(td,J=2.3,10.8Hz,1H),4.95-4.90(m,2H ),4.69(d,J=14.6Hz,1H),4.60(s,1H),4.07-3.93(m,3H),3.79(s,3H),3.75-3.66(m,2H),3.5 2(dt,J=1.9,11.6Hz,2H),2.03-1.94(m,2H),1.68-1.53(m,5H).LCMS(ESI):m / z:666.3[M+1].

[0197] Example 4

[0198]

[0199]

[0200] Compound 4B:

[0201]

[0202] Under nitrogen protection, compound 1E (100 mg, 252.99 μmol), compound 4A (97.69 mg, 379.48 μmol), Pd(dppf)Cl2.CH2Cl2 (10.33 mg, 12.65 μmol) and sodium carbonate (53.63 mg, 505.97 μmol) were dissolved in dioxane (5 ml) and water (1 ml), and the reaction solution was stirred at 80°C for 2 hours. After the reaction was completed, 0.5 ml of hydrochloric acid (2 mol) was added to the reaction solution to quench the reaction, and the mixed solution was diluted with water (10 ml) and extracted with ethyl acetate (10 ml × 2 times). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound 4B. 1 H NMR (400MHz, CDCl3) δ = 8.27 (s, 1H), 7.83 (s, 1H), 7.65 (dd, 1H, J = 1.4, 8.0Hz), 7.49 (d, 1H, J = 8.1Hz), 6.90 (d, 1H, J = 2.5Hz) ,4.90(d,1H,J=14.1Hz),4.5-4.6(m,2H),4.1-4.2(m,2H),1.6-1.6(m,3H),1.2-1.3(m,3H).LCMS(ESI):m / z:647.0[M+1].

[0203] Compound 4C:

[0204]

[0205] Under nitrogen protection, compound 4B (50 mg, 0.125 mmol), tetrahydropyran-4-amine (25.36 mg, 0.251 mmol), DIEA (48.61 mg, 0.376 mmol, 65.51 μL) were dissolved in NMP (3 mL), and the mixture was stirred at 140° C. for 4 hours. The reaction solution was diluted with ethyl acetate (10 mL) and washed with a 2M aqueous solution of hydrochloric acid (10 mL×1 time), the organic phase was extracted with ethyl acetate (10 mL×2 times), the combined organic phase was washed with brine (10 mL×1 time), and the combined organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (eluted with petroleum ether:ethyl acetate=10:1-4:1) to obtain compound 4C. 1H NMR (400MHz, CDCl3) δ = 8.07 (s, 1H), 7.77 (s, 1H), 7.6-7.7 (m, 1H), 7.4-7.5 (m, 1H), 6.26 (s, 1H), 4.87 (d, 1H, J = 13.9Hz), 4.5-4.6 (m, 2H), 4.43 (br d,1H,J=7.7Hz),4.1-4.2(m,2H),3.9-4.0(m,2H),3.7-3.9(m,1H),3.48(br t,2H,J=10.8Hz),1.9-1.9(m,2H),1.60(d,3H,J=7.3Hz),1.42(br d,2H,J=3.9Hz),1.2-1.2(m,3H).LCMS(ESI):m / z:480.1[M+1].

[0206] Compound 4D:

[0207]

[0208] To a solution of compound 4C (50 mg, 104.17 μmol) in tetrahydrofuran (5 ml) was added water and a solution of lithium hydroxide (21.86 mg, 520.86 μmol) in water (1 ml), and the reaction solution was stirred at 25°C for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, the pH of the reaction solution was adjusted to 2 with a 2-molecular hydrochloric acid aqueous solution, and then extracted with ethyl acetate (10 ml x 3 times). The combined organic phase was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude compound 4D. 1 H NMR (400MHz, CD3OD) δ = 8.06 (s, 1H), 7.86 (s, 1H), 7.79 (dd, 1H, J = 1.4, 8.0Hz), 7.6-7.7 (m, 1H), 6.59 (s, 1H), 4.94 (br s,2H),4.72(d,1H,J=14.5Hz),4.54(q,1H,J=7.3Hz),4.0-4.0(m,2H),3.5-3.6(m,2H),3.4- 3.5(m,2H),2.0-2.1(m,2H),1.7-1.7(m,3H),1.5-1.6(m,2H).LCMS(ESI):m / z:452.0[M+1].

[0209] Compound 4:

[0210]

[0211] At 0°C, DIEA (34.32 mg, 265.53 μmol, 46.25 μl) was added to a solution of compound 4D (40 mg, 88.51 μmol) and (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (21.58 mg, 97.36 μmol) in dichloromethane (4 ml), and the mixture was stirred at 0°C for 20 minutes, and then HATU (40.39 mg, 106.21 μmol) was added and continued to stir at 0°C for 40 minutes. The reaction solution was adjusted to pH <7 with aqueous hydrochloric acid (2 mol), then extracted with dichloromethane (10 ml × 2 times), and then washed with water (10 × 1 time). The organic phases were combined and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 4. 1 H NMR (400MHz, CD3OD) δ=8.06(s,1H),7.88(s,1H),7.80(dd,1H,J=1.3,8.1Hz),7.69(d,1H,J=7.9Hz),6.77(s,1H),6.70(br d,1H,J=9.9Hz),6.61(td,1H,J=2.2,10.8Hz),6.57(s,1H),4.9-5.0(m,2H),4.6-4.7(m,2H),3.9-4.1 (m,3H),3.81(s,3H),3.7-3.8(m,2H),3.5-3.6(m,2H),2.0-2.1(m,2H),1.61(d,3H,J=7.1Hz),1.56(br dd,2H,J=3.5,11.8Hz).LCMS(ESI):m / z:619.0[M+1].

[0212] Example 5

[0213]

[0214] Compound 5C:

[0215]

[0216] Under nitrogen protection, at -15°C, a solution of isopropylmagnesium bromide (1.3 mol, 3.75 ml) in tetrahydrofuran (12 ml) was slowly added with a solution of compound 5A (1 g, 4.77 mmol) in tetrahydrofuran (4 ml). The mixture was stirred at -15°C for 2 hours, and then a solution of compound 5B (1.33 g, 4.78 mmol) in tetrahydrofuran (4 ml) was added, and the reaction solution was stirred at -15°C for 2 hours. After the reaction was completed, a saturated ammonium chloride solution (20 ml) was added to quench the reaction solution, which was then diluted with water (10 ml), extracted with ethyl acetate (10 ml × 2 times), and the organic phase was washed with brine (10 ml × 1 time). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was separated by high performance liquid chromatography (formic acid system) to obtain compound 5C. 1 H NMR (400MHz, DMSO-d6) δ = 7.32 (br t, J = 4.5Hz, 2H), 7.23 (br d,J=9.5Hz,1H),5.24(d,J=5.9Hz,1H),4.36(q,J=6.2Hz,1H),3.83(dd,J=6.1,9.9Hz,1H),3.71(dd,J=7 .2,10.0Hz,1H),1.14-1.10(m,9H),0.79(s,9H),-0.07(d,J=12.1Hz,6H).LCMS(ESI):m / z:408.4[M+1].

[0217] Compound 5D:

[0218]

[0219] To a solution of compound 5C (280 mg, 686.20 μmol) in dioxane (3 ml) was added hydrochloric acid / dioxane (4 mol, 1.5 ml). The reaction mixture was stirred at 25°C for 0.5 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to obtain crude product 5D which was used directly in the next step.

[0220] Compound 5:

[0221]

[0222] At 0°C, compound 5D (55 mg, 243.28 μmol) and DIEA (81.62 mg, 631.54 μmol, 110 μl) were added to a solution of compound 1H (70 mg, 154.56 μmol) in dichloromethane (3 ml). The reaction solution was stirred at 0°C for 5 minutes, and then HATU (79 mg, 207.77 μmol) was added to continue the reaction for 2 hours. After the reaction was completed, the reaction solution was diluted with water (10 ml) and extracted with dichloromethane (10 ml × 3 times), washed with water (20 ml × 1 time), the organic phases were combined and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 5. 1 H NMR (400MHz, CD3OD) δ=8.36(s,1H),8.22(s,1H),8.15(dd,J=1.5,8.0Hz,1H),7.69(d,J=8.2Hz,1H),7.23(s,1H),7.14-7.05(m,2H),4.94(br t,J=5.8Hz,2H),4.69(d,J=14.8Hz,1H),4.63-4.57(m,1H),4.09-3.93(m,3H),3.79-3.69(m,2H),3.58-3.48(m,2H),1.99(br dd,J=1.8,12.8Hz,2H),1.68-1.55(m,5H).LCMS(ESI):m / z:624.4[M+1].

[0223] Example 6

[0224]

[0225]

[0226] Compound 6B:

[0227]

[0228] To a solution of compound 6A (5 g, 32.44 mmol) in dichloromethane (80 ml) were added copper sulfate (12.94 g, 81.10 mmol, 12.45 ml) and (S)-2-methylpropyl-2-sulfenamide (4.72 g, 38.93 mmol). The reaction solution was stirred at 50°C for 16 hours. After the reaction was completed, the reaction solution was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1-10:1 elution) to obtain compound 6B. 1H NMR (400MHz, CDCl3) δ = 8.50 (d, J = 1.0Hz, 1H), 7.20-7.15 (m, 2H), 6.77 (td, J = 2.3, 10.3Hz, 1H), 3.86 (s, 3H), 1.27 (s, 9H).

[0229] Compound 6C:

[0230]

[0231] Methylmagnesium bromide (3 mol, 17.1 ml) was added to a solution of compound 6B (5.3 g, 20.60 mmol) in tetrahydrofuran (100 ml) at -78°C under nitrogen protection, and the reaction solution was stirred at 15°C for 16 hours. After the reaction, a saturated ammonium chloride solution (30 ml) was added to the reaction solution, the organic phase was extracted with ethyl acetate (10 ml × 2 times), the combined organic phase was washed with brine (10 ml × 1 time), and the combined organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-1:1) to obtain compound 6C. LCMS (ESI): m / z: 547.2 [M+1].

[0232] Compound 6D:

[0233]

[0234] To a solution of compound 6C (3.67 g, 13.43 mmol) in dioxane (72 ml) was added hydrochloric acid / dioxane (4 mol, 36 ml). The reaction mixture was stirred at 20°C for 1 hour. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of crude product 6D which was used directly in the next step.

[0235] Compound 6F:

[0236]

[0237] Compound 6D hydrochloride (1.05 g, 5.11 mmol) and DIEA (2.37 g, 18.37 mmol, 3.2 ml) were added to a solution of compound 6E (900 mg, 4.39 mmol) in dichloromethane (20 ml) at 0°C. HATU (2.2 g, 5.79 mmol) was then added, and the reaction mixture was stirred at 0°C for 2 hours. After the reaction was complete, the reaction mixture was diluted with water (50 ml), the organic phase was extracted with dichloromethane (50 ml × 3 times), the combined organic phases were washed with brine (10 ml × 1 time), and the combined organic phases were concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (dichloromethane: methanol = 50:0-50:1) to obtain compound 6F. LCMS (ESI): m / z: 442.3 [M+1].

[0238] Compound 6G:

[0239]

[0240] Trifluoroacetic acid (1.92 g, 16.88 mmol, 1.25 ml) was added to a solution of compound 6F (300 mg, 841.78 μmol) in dichloromethane (5 ml) at 0°C. The reaction mixture was stirred at 20°C for 3 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain crude product 6G which was used directly in the next step. LCMS (ESI): m / z: 257.1 [M+1].

[0241] Compound 6I:

[0242]

[0243] DIEA (430.36 mg, 3.33 mmol, 580 μL) and compound 6H (435 mg, 1.25 mmol) were added to a solution of compound 6G (310 mg, 837.17 μmol) in acetonitrile (5 ml). The reaction solution was stirred at 80°C for 16 hours. After the reaction was completed, the reaction solution was diluted with water (20 ml), the organic phase was extracted with dichloromethane (20 ml × 3 times), the combined organic phase was washed with brine (10 ml × 1 time), and the combined organic phase was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (dichloromethane: methanol = 10:1) to obtain compound 6I. 1 H NMR(400MHz,DMSO-d6)δ=8.70(br d,J=7.9Hz,1H),8.20(d,J=1.6Hz,1H),7.91(dd,J=1.8,8.3Hz,1H),7.60 (d,J=8.3Hz,1H),6.79-6.60(m,3H),5.24(t,J=5.4Hz,1H),4.94(d,J=15. 0Hz,1H),4.89-4.83(m,1H),4.61(d,J=15.0Hz,1H),4.34(t,J=6.7Hz,1H ), 3.84-3.70 (m, 5H), 1.32 (d, J = 7.0Hz, 3H). LCMS (ESI): m / z: 489.0 [M+1].

[0244] Compound 6J:

[0245]

[0246] Under nitrogen protection, compound 6I (130 mg, 266.75 μmol), bispinacol borate, Pd(dppf)Cl2.CH2Cl2 (12 mg, 14.69 μmol), potassium tert-butoxide (80 mg, 815.16 μmol) were dissolved in dioxane (3 ml), and the reaction solution was stirred at 90°C for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure to obtain crude compound 6J, which was directly used in the next step. LCMS (ESI): m / z: 453.1 [M+1].

[0247] Compound 6K:

[0248]

[0249] Under nitrogen protection, compound 6J (140 mg, 261.97 μmol), 2,4,5-trichloropyrimidine (114 mg, 621.51 μmol, 56.29 μL), Pd(PPh3)4 (30 mg, 25.96 μmol) and sodium carbonate (70 mg, 660.44 μmol) were dissolved in dioxane (4 mL) and water (1 mL), and the reaction solution was stirred at 100°C for 1 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the filtrate was diluted with water (20 mL) and extracted with ethyl acetate (20 mL × 3 times). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain compound 6K. LCMS (ESI): m / z: 555.1 [M+1].

[0250] Compound 6:

[0251]

[0252] To a solution of compound 6K (70 mg, 126.03 μmol) in dioxane (3 ml) was added DIEA (44.52 mg, 344.48 μmol, 60 μl) and tetrahydropyran-4-amine (40 mg, 395.47 μmol). The reaction solution was stirred at 90°C for 16 hours. The reaction solution was diluted with water (20 ml) and extracted with ethyl acetate (15 ml × 3 times), the organic phase was washed with brine (30 ml × 1 time), the organic phases were combined, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 6. 1H NMR (400MHz, CD3OD) δ = 8.37 (s, 1H), 8.22 (d, J = 1.0Hz, 1H), 8.15 (dd, J = 1.6, 8.1Hz, 1H),7.69(d,J=8.1Hz,1H),6.75(s,1H),6.71-6.65(m,1H),6.54(td,J=2.2,10.7Hz ,1H),5.04(d,J=14.8Hz,1H),5.00-4.92(m,1H),4.76(d,J=14.9Hz,1H),4.47(t,J =6.6Hz,1H),4.08-3.93(m,5H),3.79(s,3H),3.53(dt,J=1.9,11.6Hz,2H),1.99(br dd, J=2.2, 12.4Hz, 2H), 1.68-1.56 (m, 2H), 1.44 (d, J=7.0Hz, 3H). LCMS (ESI): m / z: 620.5 [M+1].

[0253] Example 7

[0254]

[0255] Compound 7:

[0256]

[0257] Compound 7A (65.78 mg, 350.52 μmol) and DIEA (118.72 mg, 918.60 μmol, 160 μl) were added to a solution of compound 1H (100 mg, 220.79 μmol) in dichloromethane (3 ml) at 0°C, and the reaction solution was stirred at 0°C for 10 minutes. Then HATU (112 mg, 294.56 μmol) was added and stirring was continued for 2 hours. The reaction solution was diluted with water (10 ml) and extracted with dichloromethane (10 ml × 3 times), the organic phase was washed with brine (20 ml × 1 time), the organic phases were combined, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 7. 1H NMR (400MHz, CD3OD) δ = 8.36 (s, 1H), 8.22 (d, J = 1.1Hz, 1H), 8.15 (dd, J = 1.6, 8.1Hz, 1H) ,7.69(d,J=8.4Hz,1H),7.25-7.19(m,1H),7.16(s,1H),7.10(dd,J=7.6,15.3Hz,2H),4 .96-4.93(m,1H),4.93-4.89(m,1H),4.69(d,J=14.8Hz,1H),4.59(q,J=7.0Hz,1H),4. 10-3.93(m,3H),3.78-3.66(m,2H),3.53(dt,J=1.9,11.7Hz,2H),2.33(s,3H),1.99(br dd,J=1.9,12.6Hz,2H),1.68-1.55(m,5H).LCMS(ESI):m / z:586.1[M+1].

[0258] Example 8

[0259]

[0260] Compound 8B:

[0261]

[0262] To a solution of compound 8A (9 g, 48.39 mmol) in dichloromethane (300 ml) were added copper sulfate (19.31 g, 120.96 mmol, 18.57 ml) and (S)-2-methylpropyl-2-sulfenamide (7 g, 57.76 mmol). The reaction solution was stirred at 50°C for 48 hours. The reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 40:1-10:1) to obtain compound 8B.

[0263] Compound 8C:

[0264]

[0265] Under nitrogen protection at -78°C, a solution of allylmagnesium bromide (1 mol, 21.78 ml) in tetrahydrofuran (40 ml) was added with a solution of compound 8B (3 g, 10.37 mmol) in tetrahydrofuran (20 ml), and the reaction solution was stirred at -78°C for 1 hour. After the reaction, the mixture was quenched with a saturated ammonium chloride solution (30 ml), then diluted with water (50 ml), extracted with ethyl acetate (50 ml × 3 times), the organic phase was washed with brine (50 ml × 1 time), and the organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was separated by high performance liquid chromatography (formic acid system) to obtain compound 8C. LCMS (ESI): m / z: 319.0 [M+1].

[0266] Compound 8D:

[0267]

[0268] Compound 8C (1 g, 3.15 mmol) was dissolved in dichloromethane (10 ml) and methanol (10 ml) at -78°C under ozone (3.15 mmol, 15 psi), sodium borohydride (240.00 mg, 6.34 mmol) was added to the reaction solution, and stirred at -78°C for 3 hours. After the reaction, a saturated ammonium chloride solution (30 ml) was added to quench the reaction solution, which was then diluted with water (30 ml), extracted with ethyl acetate (50 ml × 2 times), and the organic phase was washed with brine (80 ml × 1 time). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain compound 8D. LCMS (ESI): m / z: 337.0 [M+1].

[0269] Compound 8E:

[0270]

[0271] Compound 8D (100.00 mg, 311.30 μmol) was dissolved in dimethylamine (2 mol, 3 ml) and reacted at 120°C for 4 hours under microwave conditions. After the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (eluted with dichloromethane: methanol = 10:1) to obtain compound 8E. LCMS (ESI): m / z: 286.1 [M+1].

[0272] Compound 8F:

[0273]

[0274] To a solution of compound 8E (90 mg, 315.34 μmol) in dioxane (2 ml) was added hydrochloric acid / dioxane (4 mol, 1 ml). The reaction mixture was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude hydrochloride of compound 8F which was used directly in the next step. 1 H NMR (400MHz, DMSO-d6) δ = 8.33 (br s, 3H), 7.57 (brt, J = 7.9Hz, 1H), 6.73-6.58 (m, 2H), 4.26 (br d, J = 1.6Hz, 1H), 3.84-3.66 (m, 2H), 3.07 (s, 6H).

[0275] Compound 8:

[0276]

[0277] At 0°C, compound 8F (54.00 mg, 248.06 μmol, HCl) and DIEA (118.72 mg, 918.60 μmol, 160 μl) were added to a solution of compound 1H (100 mg, 220.79 μmol) in dichloromethane (3 ml). The reaction solution was stirred for 10 minutes, and HATU (110 mg, 289.30 μmol) was added and continued to stir for 2 hours. The reaction solution was diluted with water (10 ml) and extracted with dichloromethane (10 ml × 3 times). The organic phase was washed with brine (20 ml × 1), the combined organic phases were filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system), and then the free base of compound 8 was obtained by chiral separation method (0.1% ammonia methanol system, retention time 3.6 minutes, ee value: 100%). 1 H NMR (400MHz, MeOH-d4) δ = 8.37 (s, 1H), 8.22 (d, J = 1.1Hz, 1H), 8.16 (dd, J = 1.6, 8.1Hz, 1H), 7.70 (d, J = 8 .1Hz,1H),7.45(dd,J=7.3,8.5Hz,1H),6.56(d,J=7.2Hz,1H),6.49(d,J=8.4Hz,1H),4.92-4.89(m,1H ),4.81-4.65(m,2H),4.60(q,J=7.1Hz,1H),4.12-3.93(m,3H),3.80(dq,J=5.6,10.9Hz,2H),3.53(dt ,J=1.9,11.6Hz,2H),2.91(s,6H),2.04-1.95(m,2H),1.66-1.56(m,5H).LCMS(ESI):m / z:616.3[M+1].

[0278] Example 9

[0279]

[0280] Compound 9C:

[0281]

[0282] Compound 9A (2 g, 14.58 mmol, 1.89 ml) was dissolved in methanol (2 ml), and compound 9B (2.08 g, 13.13 mmol, 1.85 ml) was added, and the mixture was stirred at 10° C. for 16 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (eluted with petroleum ether: ethyl acetate = 4:1-2:1) to obtain compound 9C. 1 H NMR (400MHz, CDCl3) δ = 7.24 (d, J = 8.5Hz, 2H), 6.92-6.79 (m, 2H), 3.80 (s, 3H), 3.75 (s, 2H), 3.69 (s, 6H), 3.44 (quin, J = 6.2Hz, 1H), 2.59 (d, J = 6.3Hz, 4H).

[0283] Compound 9D:

[0284]

[0285] To a solution of compound 9C (2.85 g, 9.65 mmol) in tetrahydrofuran (30 ml) were added (Boc)2O (2.53 g, 11.57 mmol, 2.66 ml) and DIEA (1.77 g, 13.68 mmol, 2.38 ml). The mixture was stirred at 10°C for 16 hours. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (eluted with petroleum ether: ethyl acetate = 10:1-5:1) to obtain compound 9D. 1 H NMR (400MHz, CDCl3) δ = 7.22 (br s, 2H), 6.85 (d, J = 8.6Hz, 2H), 4.42 (br s,2H),4.35-4.20(m,1H),3.85-3.73(m,3H),3.61(s,6H),2.86-2.50(m,4H),1.50(br d,J=16.0Hz,9H). LCMS(ESI):m / z:296.1[M+1].

[0286] Compound 9E:

[0287]

[0288] Under nitrogen protection, LiAlD4 (910 mg, 23.98 mmol, 1.24 ml) was added to a solution of compound 9D (3.16 g, 7.99 mmol) in tetrahydrofuran (35 ml). The reaction solution was stirred at 0°C for 20 minutes. After the reaction was completed, 15% aqueous sodium hydroxide (0.91 ml) was added to the mixture to quench, and then filtered. The filtrate was concentrated under reduced pressure to obtain crude product 9E which was directly used in the next step. 1 H NMR (400MHz, CDCl3) δ = 7.21 (br d, J = 7.4Hz, 2H), 6.88-6.82 (m, 2H), 4.52-4.32 (m, 1H), 4.26 (br s, 2H), 3.80 (s, 3H), 1.67 (br d,J=6.9Hz,4H),1.50-1.46(m,9H).

[0289] Compound 9F:

[0290]

[0291] Potassium tert-butoxide (975.00 mg, 8.69 mmol) was added to a solution of compound 9E (1.5 g, 4.37 mmol) in tetrahydrofuran (20 ml), and after stirring for 15 minutes, MsCl (555.00 mg, 4.85 mmol, 375.00 μL) and potassium tert-butoxide (495.00 mg, 4.41 mmol) were added. The mixture was stirred at 25°C for 2 hours. After the reaction was completed, a saturated ammonium chloride solution (10 ml) was added to the reaction solution, and then extracted with ethyl acetate (50 ml x 3 times). The organic phases were combined and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-5:1) to obtain compound 9F. 1 H NMR (400MHz, DMSO-d6) δ = 7.15 (d, J = 8.7Hz, 2H), 6.87 (d, J = 8.7Hz, 2H), 4.29 (br s, 2H), 4.08-3.75 (m, 1H), 3.71 (s, 3H), 1.63 (br t,J=12.3Hz,2H),1.46-1.30(m,11H)

[0292] Compound 9G:

[0293]

[0294] To a solution of compound 9F (560 mg, 1.72 mmol) in methanol (2 ml) was added hydrochloric acid / dioxane (4 mol, 2 ml). The reaction mixture was stirred at 25°C for 2 hours. After the reaction was completed, the reaction mixture was poured into methyl tert-butyl ether (10 ml), filtered to obtain a filter cake, which was dissolved in water (10 ml), adjusted to pH = 11 with sodium hydroxide (2 mol) aqueous solution, and then extracted with ethyl acetate (10 ml x 2 times). The organic phases were combined and concentrated under reduced pressure to obtain a crude product 9G which was used directly in the next step. 1 H NMR (400MHz, CDCl3) δ = 7.25 (d, J = 8.6Hz, 2H), 6.90-6.83 (m, 2H), 3.81 (s, 3H), 3 .78(s,2H),2.73(tt,J=4.1,10.5Hz,1H),1.84(dd,J=4.1,13.3Hz,2H),1.45(br d,J=11.7Hz,2H).

[0295] Compound 9H:

[0296]

[0297] Palladium / carbon (50 mg, 10% purity) was added to a solution of compound 9G (150 mg, 665.72 μmol) in methanol (10 ml). The reaction solution was stirred at 50°C for 16 hours under hydrogen (0.8 MPa). After the reaction was complete, the mixture was filtered, the filtrate was concentrated under reduced pressure and then dissolved in methanol (1 ml), the pH of the mixture was adjusted to 1 with hydrochloric acid / dioxane (4 mol), and then methyl tert-butyl ether (5 ml) was added to the mixture and filtered to obtain crude product 9H which was directly used in the next step. 1 H NMR (400MHz, DMSO-d6) δ = 8.11 (br s, 2H), 3.29-3.15 (m, 1H), 1.82 (dd, J = 4.2, 13.0Hz, 2H), 1.52 (br t, J = 12.0Hz, 2H).

[0298] Compound 9I:

[0299]

[0300] To a solution of compound 1F (1.18 g, 2.83 mmol) in tetrahydrofuran (5 ml) and water (5 ml) was added lithium hydroxide monohydrate (472.00 mg, 11.25 mmol). The reaction solution was stirred at 0°C for 1 hour. After the reaction was completed, water (10 ml) was added to the reaction solution, and then extracted with ethyl acetate (10 ml × 1 time), the aqueous phase was adjusted to pH = 2 with hydrochloric acid (2 mol), and extracted with dichloromethane (10 ml × 2 times), the organic phases were combined and filtered, and concentrated under reduced pressure to obtain crude product 9I which was directly used in the next step. 1 H NMR (400MHz, CDCl3) δ = 8.73 (s, 1H), 8.42 (d, J = 1.3Hz, 1H), 8.20 (dd, J = 1.5, 8.1Hz, 1H ),7.59(d,J=8.1Hz,1H),4.94-4.86(m,1H),4.70-4.60(m,2H),1.72(d,J=7.3Hz,3H). LCMS(ESI):m / z:387.9[M+1].

[0301] Compound 9J:

[0302]

[0303] At 0°C, HATU (585.85 mg, 1.54 mmol), (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (280 mg, 1.26 mmol) and DIEA (406.27 mg, 3.14 mmol, 547.54 μL) were added to a solution of compound 9I (450 mg, 1.04 mmol, 89.44% purity) in dichloromethane (10 mL). The reaction solution was stirred at 15°C for 30 minutes. The reaction solution was diluted with water (2 mL) and extracted with ethyl acetate (2 mL × 3 times). The organic phases were combined and filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (eluted with petroleum ether: ethyl acetate = 2:1) to obtain compound 9F. 1 H NMR (400MHz, CDCl3) δ=8.68-8.64(m,2H),8.39-8.32(m,2H),8.19-8.12(m,2H),7.56-7.44(m,2H),6.60-6.24(m, 7H),4.99-4.90(m,2H),4.74-4.67(m,1H),4.56-4.50(m,3H),3.70(s,3H),1.97-1.92(m,3H),1.57-1.50(m,7H). LCMS(ESI):m / z:555.0[M+1].

[0304] Compound 9:

[0305]

[0306] To a solution of compound 9J (90 mg, 162.04 μmol) and 9H (70 mg, 393.05 μmol) in dioxane (3 ml) was added DIEA (83.85 mg, 648.75 μmol, 113 μl). The reaction solution was stirred at 100°C for 16 hours. The reaction solution was diluted with water (5 ml) and extracted with ethyl acetate (2 ml x 3 times). The combined organic phases were dried, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 9. 1 H NMR (400MHz, MeOH-d4) δ = 8.36 (s, 1H), 8.22 (s, 1H), 8.15 (dd, J = 1.5, 8.1Hz, 1H), 7.69 (d, J = 8.3Hz, 1H), 6.75 (s, 1H), 6.68 (br d,J=9.3Hz,1H),6.59(td,J=2.2,10.7Hz,1H),4.92(d,J=7.1Hz,1H),4.83-4.79(m,1H),4.70(d,J=15.0Hz,1H),4.59 (q,J=7.1Hz,1H),4.10-3.97(m,1H),3.79(s,3H),3.76-3.69(m,2H),1.98(dd,J=4.1,13.4Hz,2H),1.63-1.55(m,5H). LCMS(ESI):m / z:624.0[M+1].

[0307] Example 10

[0308]

[0309]

[0310] Compound 10A:

[0311]

[0312] Under nitrogen protection, compound 1C (400 mg, 1.61 mmol), ethyl-2-bromoacetate (404 mg, 2.42 mmol, 267.55 μL), potassium carbonate (450 mg, 3.26 mmol) were dissolved in DMF (6 mL), and the reaction solution was stirred at 25°C for 16 hours. After the reaction was completed, it was diluted with water (30 mL) and extracted with ethyl acetate (20 mL × 3 times). The organic phase was washed with brine (50 mL × 1 time), and the combined organic phases were concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-5:1) to obtain compound 10A. 1H NMR (400MHz, CDCl3) δ = 7.96 (d, J = 1.6Hz, 1H), 7.74 (dd, J = 1.8, 8.3Hz, 1H), 7.30 (d, J = 8.2Hz, 1H), 4. 59 (s, 2H), 4.24 (q, J = 7.1Hz, 2H), 4.08 (s, 2H), 1.30 (t, J = 7.2Hz, 3H). LCMS (ESI): m / z: 320.0 [M+1].

[0313] Compound 10B:

[0314]

[0315] Under nitrogen protection, compound 10A (360 mg, 1.08 mmol), bispinacol borate (410 mg, 1.61 mmol), Pd(dppf)Cl2.CH2Cl2 (45 mg, 55.10 μmol) and potassium acetate (320 mg, 3.26 mmol) were dissolved in dioxane (4 ml), and the reaction solution was stirred at 90°C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain crude compound 10B, which was directly used in the next step. LCMS (ESI): m / z: 300.1 [M+1].

[0316] Compound 10C:

[0317]

[0318] Under nitrogen protection, compound 10B (322 mg, 1.08 mmol), 2,4,5-trichloropyrimidine (355 mg, 1.94 mmol), Pd(PPh3)4 (62 mg, 53.65 μmol) and sodium carbonate (235 mg, 2.22 mmol) were dissolved in dioxane (6 ml) and water (1.5 ml), and the reaction solution was stirred at 90°C for 2 hours. After the reaction, the reaction solution was filtered, the filtrate was diluted with water (20 ml) and extracted with ethyl acetate (20 ml × 3 times). The organic phase was washed with brine (30 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1 elution) to obtain compound 10C. LCMS (ESI): m / z: 374.0 [M+1].

[0319] Compound 10D:

[0320]

[0321] Tetrahydropyran-4-amine (140 mg, 1.38 mmol, 694.34 μL) and DIEA (148.40 mg, 1.15 mmol, 200 μL) were added to compound 10C (185 mg, 459.91 μmol) in dioxane (4 mL), and the reaction solution was stirred at 90°C for 16 hours. Water (10 mL) and ammonium chloride (20 mL) were added to the reaction solution, and then extracted with ethyl acetate (20 mL × 3 times). The organic phase was washed with brine (20 mL × 1 time), dried, filtered, and concentrated under reduced pressure to obtain crude product 10D, which was directly used in the next step. LCMS (ESI): m / z: 467.1 [M+1].

[0322] Compound 10E:

[0323]

[0324] To a solution of compound 10D (140 mg, 299.83 μmol) in tetrahydrofuran (2 ml) and ethanol (1 ml) was added a solution of lithium hydroxide monohydrate (20 mg, 476.64 μmol) in water (1 ml), and the reaction solution was stirred at 20°C for 0.5 hours. The reaction solution was diluted with water (10 ml) and extracted with ethyl acetate (10 ml x 1 time), and the aqueous phase was added with hydrochloric acid (2 mol) aqueous solution to adjust pH = 2, and then extracted with ethyl acetate (10 ml x 3 times). The combined organic phases were dried, filtered, and concentrated under reduced pressure to obtain a crude product 10E which was directly used in the next step. LCMS (ESI): m / z: 438.9 [M+1].

[0325] Compound 10:

[0326]

[0327] To a solution of compound 10E (120 mg, 273.42 μmol) in dichloromethane (4 ml) were added (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (90 mg, 406.03 μmol) and DIEA (148.40 mg, 1.15 mmol, 200 μl) at 0°C, and the reaction solution was stirred at 0°C for 10 minutes. Then HATU (142 mg, 373.46 μmol) was added and the reaction was continued for 2 hours. The reaction solution was diluted with water (10 ml) and extracted with ethyl acetate (10 ml × 3 times). The organic phase was washed with brine (20 ml × 1 time), dried, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (formic acid system) to obtain the free base of compound 10. 1H NMR (400MHz, CD3OD) δ=8.36(s,1H),8.24(d,J=1.1Hz,1H),8.15(dd,J=1.5,8.2Hz,1H),7.68(d,J=8.2Hz,1H),6.77(s,1H),6.71(br d,J=9.4Hz,1H),6.58(td,J=2.3,10.8Hz,1H),4.99(dd,J=5.3,6.9Hz,1H),4.79-4.6 2(m,2H),4.19-3.93(m,5H),3.83-3.68(m,5H),3.53(dt,J=2.0,11.7Hz,2H),1.99(br dd,J=2.1,12.4Hz,2H),1.68-1.55(m,2H).LCMS(ESI):m / z:606.1[M+1].

[0328] Embodiment 11

[0329]

[0330] Compound 11B:

[0331]

[0332] Chlorosulfonic acid (61.32 g, 526.21 mmol, 35.04 ml) was slowly added to a solution of compound 11A (30 g, 175.40 mmol, 21.58 ml) in dichloromethane (20 ml) at 0°C, and the mixture was stirred at 20°C for 3 hours. After the reaction was complete, the reaction solution was slowly added to ice water, and then extracted with dichloromethane (100 ml × 2 times), and the organic phases were combined and washed with water (100 ml × 2 times), dried, filtered, and concentrated under reduced pressure to obtain a crude product 11B, which was directly used in the next step. 1 HNMR (CDCl3, 400MHz) δ8.22 (d, 1H, J = 2.1Hz), 7.75 (dd, 1H, J = 2.0, 8.2Hz), 7.33 (d, 1H, J = 8.2Hz), 2.76 (s, 3H).

[0333] Compound 11C:

[0334]

[0335] To a solution of compound 11B (4 g, 14.84 mmol) in dichloromethane (100 ml) were added NBS (2.91 g, 16.32 mmol) and AIBN (24.37 mg, 148.40 μmol), and the reaction solution was stirred at 80°C for 3 hours. After the reaction, the reaction solution was filtered and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (eluted with petroleum ether) to obtain compound 11C. 1 H NMR (CDCl3, 400MHz) δ8.23 (d, 1H, J = 2.1Hz), 7.86 (dd, 1H, J = 2.0, 8.3Hz), 7.63 (d, 1H, J = 8.4Hz), 4.94 (s, 2H).

[0336] Compound 11D:

[0337]

[0338] To a solution of compound 11C (1 g, 2.87 mmol) and compound (R)-tert-butyl-2-aminopropyl ester (521.35 mg, 2.87 mmol) in acetonitrile (10 ml) was added a solution of sodium carbonate (912.55 mg, 8.61 mmol) in water (2 ml), and the mixture was stirred at 20°C for 1 hour, and then stirred at 80°C for 13 hours. After the reaction was complete, the reaction solution was diluted with water (20 ml) and extracted with ethyl acetate (20 ml × 3 times). The organic phase was washed with brine (20 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude compound 11D which was directly used in the next step. 1 H NMR (CDCl3, 400MHz) δ7.94 (s, 1H), 7.72 (d, 1H, J = 8.2Hz), 7.29 (d, 1H, J = 8.2Hz) ), 4.86 (d, 1H, J = 13.9Hz), 4.4-4.5 (m, 2H), 1.61 (d, 3H, J = 7.3Hz), 1.45 (s, 9H). LCMS(ESI):m / z:377.9[M+1].

[0339] Compound 11E:

[0340]

[0341] Under nitrogen protection, compound 11D (15 g, 39.87 mmol), bispinacol borate (15.19 g, 59.80 mmol), Pd(dppf)Cl2.CH2Cl2 (1.63 g, 1.99 mmol) and potassium acetate (7.82 g, 79.73 mmol) were dissolved in dioxane (70 ml), and the reaction solution was stirred at 90°C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain crude compound 11E, which was directly used in the next step. LCMS (ESI): m / z: 341.7 [M+1].

[0342] Compound 11F:

[0343]

[0344] Under nitrogen protection, compound 11E (4.2 g, 12.31 mmol), 2,4,5-trichloropyrimidine (4.52 g, 24.62 mmol), Pd(PPh3)4 (711.25 mg, 615.50 μmol) and sodium carbonate (2.61 g, 24.62 mmol) were dissolved in dioxane (20 ml) and water (4 ml), and the reaction solution was stirred at 90°C for 16 hours. After the reaction, the reaction solution was filtered, the filtrate was diluted with water (30 ml) and extracted with ethyl acetate (30 ml × 3 times). The organic phase was washed with brine (30 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-3:1 elution) to obtain compound 11F. 1 H NMR (CDCl3, 400MHz) δ8.71 (s, 1H), 8.41 (d, 1H, J = 1.1Hz), 8.18 (dd, 1H, J = 1.5, 8.1Hz), 7.58 (d, 1H, J = 8.1Hz) ,5.01(d,1H,J=14.5Hz), 4.62(d,1H,J=14.5Hz), 4.48(d,1H,J=7.3Hz), 1.64(d,3H,J=7.3Hz), 1.46(s,9H).

[0345] Compound 11H:

[0346]

[0347] Compound 11F (1.3 g, 2.93 mmol) in dioxane (4 ml) was added with compound 11g (682.86 mg, 4.39 mmol) and DIEA (1.89 g, 14.63 mmol, 2.55 ml), and the reaction solution was stirred at 100°C for 16 hours, and the reaction solution was diluted with ethyl acetate (30 ml) and washed with 1 molar hydrochloric acid (30 ml × 1 time) aqueous solution, and the aqueous phase was extracted with ethyl acetate (30 ml × 3 times), and the combined organic phase was washed with brine (30 ml × 1 time), dried, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-2:1) to obtain compound 11H. 1 H NMR (CDCl3, 400MHz) δ8.2-8.3(m,1H),8.18(br s,1H),7.99(dd,1H,J=1.4,8.1Hz),7.44(d,1H,J=8.3Hz),5.47(br d,1H,J=9.0Hz),4.91(d,1H,J=14.4Hz),4.6-4.7(m,1H),4.52(d,1H,J=14.3Hz),4.40(q,1H,J=7.3Hz),4.1-4.3(m,2H),3.99(br dd,1H,J=4.4,11.6Hz),3.4-3.6(m,2H),1.9-2.0(m,1H),1.8-1.9(m,1H),1.56(d,3H,J=7.4Hz),1.38(s,9H), LCMS(ESI):m / z:526.9[M+1].

[0348] Compound 11I:

[0349]

[0350] Trifluoroacetic acid (1.84 g, 16.13 mmol, 1.19 ml) was added to a solution of compound 11H (850 mg, 1.61 mmol) in dichloromethane (10 ml), and the reaction solution was stirred at 50°C for 16 hours. After the reaction was complete, the reaction solution was diluted with water (30 ml) and extracted with ethyl acetate (30 ml × 2 times). The organic phase was washed with brine (30 ml × 1 time), dried, filtered, and concentrated under reduced pressure to give a crude compound 11I which was directly used in the next step. LCMS (ESI): m / z: 470.8 [M+1].

[0351] Compound 11:

[0352]

[0353] At -5°C, (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (361.3 mg, 1.63 mmol) and DIEA (526.65 mg, 4.07 mmol, 200 μL) were added to a solution of compound 11I (639.6 mg, 1.36 mmol) in dichloromethane (10 ml), and the reaction solution was stirred at 0°C for 10 minutes. Then HATU (774.70 mg, 2.04 mmol) was added and the reaction was continued for 1 hour. The reaction solution was diluted with water (10 ml) and extracted with dichloromethane (10 ml × 3 times). The organic phase was washed with brine (20 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (trifluoroacetic acid system) to obtain the free base of compound 11. 1 H NMR(CD3OD,400MHz)δ8.43(s,1H),8.25(s,1H),8.17(dd,1H,J=1.5,8.2Hz),7.72(d,1H,J=8.1H z),6.77(s,1H),6.7-6.7(m,1H),6.61(td,1H,J=2.3,10.8Hz),4.9-4.9(m,1H),4.7-4.7(m,1H), 4.6-4.6(m,1H),4.2-4.4(m,1H),4.1-4.2(m,1H),4.0-4.1(m,1H),3.81(s,3H),3.7-3.8(m,2H), 3.4-3.6(m,2H),2.0-2.1(m,1H),1.7-1.9(m,1H),1.5-1.7(m,3H).LCMS(ESI):m / z:684.1[M+1].

[0354] Example 12

[0355]

[0356] Compound 12A:

[0357]

[0358] Under nitrogen protection, compound 11E (14.8 g, 43.38 mmol), 2,4-dichloropyrimidine-5-amine (14.23 g, 86.76 mmol), Pd(PPh3)4 (2.51 g, 2.17 mmol) and sodium carbonate (9.2 g, 86.76 mmol) were dissolved in dioxane (50 ml) and water (10 ml), and the reaction solution was stirred at 90°C for 16 hours. After the reaction, the reaction solution was filtered, the filtrate was diluted with water (100 ml) and extracted with ethyl acetate (100 ml × 2 times). The organic phase was washed with brine (100 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1-3:1 elution) to obtain compound 12A. LCMS (ESI): m / z: 424.8 [M+1].

[0359] Compound 12B:

[0360]

[0361] To a solution of cuprous bromide (50.64 mg, 353.02 μmol, 10.75 μL) and tert-butyl nitrite (60.67 mg, 588.37 μmol, 69.98 μL) in acetonitrile (3 mL) was added a solution of compound 12A (100 mg, 235.35 μmol) in acetonitrile (3 mL), and the reactants were stirred at 20°C for 18 hours. The reaction solution was diluted with water (10 mL) and extracted with ethyl acetate (10 mL × 2 times). The organic phase was washed with brine (10 mL × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1 elution) to obtain compound 12B. 1 H NMR (CDCl3, 400MHz) δ8.84 (s, 1H), 8.35 (d, 1H, J = 1.1Hz), 8.11 (dd, 1H, J = 1.6, 8.1Hz), 7.57 (d, 1H, J = 8.4Hz), 5.01 (d, 1H, J = 14.5Hz), 4.62 (d, 1H, J = 14.5Hz), 4.49 (d, 1H, J = 7.4Hz), 1.64 (d, 3H, J = 7.4Hz), 1.46 (s, 9H). LCMS (ESI): m / z: 489.8 [M+1]. Compound 12C:

[0362]

[0363] Compound 9H (869.3 mg, 6.14 mmol) and DIEA (1.19 g, 9.21 mmol, 2.55 ml) were added to a solution of compound 12B (1.5 g, 3.07 mmol) in dioxane (15 ml), and the reaction solution was stirred at 90°C for 16 hours. The reaction solution was diluted with ethyl acetate (30 ml) and washed with a 1 molar hydrochloric acid aqueous solution (30 ml × 1 time), the aqueous phase was extracted with ethyl acetate (30 ml × 3 times), the combined organic phase was washed with brine (30 ml × 1 time), dried, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-2:1) to obtain compound 12C. 1 H NMR (CDCl3, 400MHz) δ8.66 (d, 1H, J = 5.1Hz), 8.4-8.5 (m, 1H), 8.36 (dd, 1H, J = 1.6, 8.1Hz), 7.63 (d, 1H, J = 5.1Hz), 7 .5-7.5(m,1H),4.9-5.0(m,1H),4.54(d,1H,J=14.6Hz),4.40(q,1H,J=7.3Hz),1.56(d,3H,J=7.3Hz),1.38(s,9H). LCMS(ESI):m / z:559.1[M+1].

[0364] Compound 12D:

[0365]

[0366] Trifluoroacetic acid (1.23 g, 10.76 mmol, 796.86 ml) was added to a solution of compound 12C (600 mg, 1.08 mmol) in dichloromethane (10 ml), and the reaction solution was stirred at 50°C for 6 hours. After the reaction was complete, the reaction solution was diluted with water (30 ml) and extracted with ethyl acetate (30 ml x 2 times). The organic phase was washed with brine (30 ml x 1 time), dried, filtered, and concentrated under reduced pressure to give a crude compound 12D which was used directly in the next step. LCMS (ESI): m / z: 502.19 [M+1].

[0367] Compound 12:

[0368]

[0369] To a solution of compound 12D (400 mg, 797.79 μmol) in dichloromethane (10 ml) were added (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (212.2 mg, 957.34 μmol) and DIEA (309.32 mg, 2.39 mmol, 416.87 μl) at -5°C, and the reaction solution was stirred at 0°C for 10 minutes. HATU (455.01 mg, 1.2 mmol) was then added and the reaction was continued for 1 hour. The reaction solution was diluted with water (10 ml) and extracted with dichloromethane (10 ml × 3 times). The organic phase was washed with brine (20 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (trifluoroacetic acid system) to obtain the free base of compound 12. 1 H NMR (CD3OD, 400MHz) δ8.48 (s, 1H), 8.17 (s, 1H), 8.0-8.1 (m, 1H), 7.70 (d, 1H, J = 8.1Hz), 6.77 (s, 1H), 6.70 (br d,1H,J=9.5Hz),6.6-6.7(m,1H),5.0-5.0(m,1H),4.69(s,1H),4.6-4.6(m,1H),4.0-4.1(m,1H),3.81 (s,3H),3.7-3.8(m,2H),1.99(dd,2H,J=4.2,13.3Hz),1.5-1.7(m,6H).LCMS(ESI):m / z:670.0[M+1].

[0370] Embodiment 13

[0371]

[0372]

[0373] Compound 13A:

[0374]

[0375] Compound 11G (955.37 mg, 6.14 mmol) and DIEA (1.98 g, 15.35 mmol, 2.67 ml) were added to a solution of compound 12B (1.5 g, 3.07 mmol) in dioxane (10 ml), and the reaction solution was stirred at 90°C for 16 hours. The reaction solution was diluted with ethyl acetate (30 ml) and washed with a 1 molar aqueous solution of hydrochloric acid (30 ml × 1 time), the aqueous phase was extracted with ethyl acetate (30 ml × 3 times), the combined organic phase was washed with brine (30 ml × 1 time), dried, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1-2:1 elution) to obtain compound 13A. LCMS (ESI): m / z: 572.9 [M+1]. Compound 13B:

[0376]

[0377] Trifluoroacetic acid (1.2 g, 10.5 mmol, 777.39 ml) was added to a solution of compound 13A (1.2 g, 2.1 mmol) in dichloromethane (15 ml), and the reaction solution was stirred at 50°C for 16 hours. After the reaction was complete, the reaction solution was diluted with water (30 ml) and extracted with ethyl acetate (30 ml × 2 times). The organic phase was washed with brine (30 ml × 1 time), dried, filtered, and concentrated under reduced pressure to give a crude compound 13B which was directly used in the next step. LCMS (ESI): m / z: 516.9 [M+1].

[0378] Compound 13:

[0379]

[0380] At -5°C, (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (464.51 mg, 2.1 mmol) and DIEA (677.12 mg, 5.24 mmol, 912.56 μl) were added to a solution of compound 13B (900 mg, 1.75 mmol) in dichloromethane (20 ml), and the reaction solution was stirred at -5°C for 10 minutes. Then HATU (455.01 mg, 1.2 mmol) was added and the reaction was continued for 1 hour. The reaction solution was diluted with water (10 ml) and extracted with dichloromethane (20 ml × 3 times). The organic phase was washed with brine (20 ml × 1 time), dried, filtered and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (trifluoroacetic acid system) to obtain the free base of compound 13. 1H NMR (CD3OD, 400MHz) δ8.52(s,1H),8.18(s,1H),8.10(dd,1H,J=1.5,8.1Hz),7.71(d,1H,J=8.1Hz),6.77(s,1H),6.70(br d,1H,J=9.3Hz),6.61(td,1H,J=2.1,10.7Hz),5.0-5.0(m,2H),4.7-4.8(m,2H),4.61(q,1H,J=7.1Hz),4.2-4.4(m,1H),4.12(br t,1H,J=12.3Hz),4.0-4.1(m,1H),3.81(s,3H),3.7-3.8(m,2H),3.5-3.7(m,2H),2.05(dq,1H,J=4.3,12.6Hz),1.83(br dd, 1H, J = 3.5, 13.3Hz), 1.61 (d, 3H, J = 7.1Hz). LCMS(ESI):m / z:684.1[M+1].

[0381] Embodiment 14

[0382]

[0383] Compound 14A:

[0384]

[0385] Sodium borodeuteride (646.80 mg, 17.10 mmol) was added to a tetrahydrofuran (10 ml) solution of compound 1B (1.4 g, 5.34 mmol), and then the temperature of the reaction solution was lowered to -5°C, and boron trifluoride ether (2.43 g, 17.09 mmol, 2.11 ml) was slowly added dropwise to the reaction solution, and the mixture was stirred at 80°C for two hours. After the reaction was completed, the reaction solution was quenched with an aqueous ammonium chloride solution (20 ml) at 0°C, and then ethyl acetate (60 ml) was added and filtered, and the filtrate was extracted with ethyl acetate (20 ml × 2 times), and the combined organic phase was washed with brine (30 ml × 1 time), dried, and concentrated under reduced pressure to obtain a crude product. The crude product was stirred at 25°C for 0.5 hours with petroleum ether: ethyl acetate = 3:1, and the filter cake obtained by filtration was compound 14A. 1 HNMR (400MHz, DMSO-d6) δ = 8.12 (d, J = 1.3Hz, 1H), 7.95 (s, 1H), 7.87 (dd, J = 1.3, 8.2Hz, 1H), 7.54 (d, J = 8.3Hz, 1H). LCMS(ESI):m / z:250.11.

[0386] Compound 14B:

[0387]

[0388] Under nitrogen protection, compound 14A (990 mg, 3.60 mmol), ethyl 2-bromo-(2S) propionate (990 mg, 5.14 mmol) and potassium carbonate (990 mg, 5.47 mmol) were dissolved in DMF (10 ml), and the mixture was stirred at 20°C for 16 hours. The reaction solution was diluted with 30 ml of water and then extracted with ethyl acetate (30 ml x 2 times). The combined organic phase was washed with water (40 ml x 2 times) and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 20:1-15:1) to obtain compound 14B. 1 H NMR (400MHz, CDCl3, 400MHz) δ = 7.86 (d, J = 1.7Hz, 1H), 7.65 (dd, J = 1.7, 8.3Hz, 1H), 7.20 (d, J = 8.7Hz, 1H), 4.47 ( q, J=7.3Hz, 1H), 4.17-4.07 (m, 2H), 1.56 (d, J=7.3Hz, 3H), 1.19 (t, J=7.1Hz, 3H). LCMS (ESI): m / z: 350.22[M+1].

[0389] Compound 14C:

[0390]

[0391] Under nitrogen protection, compound 14B (550 mg, 1.57 mmol), bispinacol borate (630 mg, 2.48 mmol), Pd(dppf)Cl2.CH2Cl2 (80 mg, 97.96 μmol) and potassium acetate (314 mg, 3.2 mmol) were dissolved in dioxane (8 ml), and the reaction solution was stirred at 100°C for 2 hours. The reaction solution was filtered and concentrated under reduced pressure to obtain crude compound 14C, which was directly used in the next step. LCMS (ESI): m / z: 315.15 [M+1].

[0392] Compound 14D:

[0393]

[0394] Under nitrogen protection, compound 14C (494 mg, 1.57 mmol), 2,4-dichloropyrimidine-5-amine (462 mg, 2.82 mmol), Pd(PPh3)4 (96 mg, 83.08 μmol) and sodium carbonate (334 mg, 3.15 mmol) were dissolved in dioxane (5 ml) and water (1 ml), and the reaction solution was stirred at 100° C. for 2 hours. After the reaction was completed, the reaction solution was filtered, dried, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1-1:1) to obtain compound 14D. 1 H NMR (CDCl3, 400MHz) δ = 8.20 (d, J = 1.1Hz, 1H), 8.13 (s, 1H), 8.00 (dd, J = 1.4, 8.0Hz, 1H), 7.51 (d, J = 8.0Hz, 1H), 4. 51(q,J=7.3Hz,1H),4.19-4.09(m,2H),1.59(d,J=7.4Hz,3H),1.24-1.17(m,3H).LCMS(ESI):m / z:398.86[M+1].

[0395] Compound 14E:

[0396]

[0397] At 0°C, a solution of compound 14D (500 mg, 1.25 mmol) in acetonitrile (3 ml) was added to a solution of cuprous bromide (360 mg, 2.51 mmol, 76.43 μL) and tert-butyl nitrite (387 mg, 3.75 mmol, 446.37 μL) in acetonitrile (3 ml), and the reactants were stirred at 25°C for 12 hours. The reaction solution was diluted with water (10 ml) and extracted with ethyl acetate (10 ml × 3 times). The organic phase was washed with brine (10 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1-3:1 elution) to obtain compound 14E. 1 H NMR (CDCl3, 400MHz) δ = 8.76 (s, 1H), 8.27 (d, J = 1.1Hz, 1H), 8.04 (dd, J = 1.7, 8.1Hz, 1H), 7.50 (d, J =8.1Hz, 1H), 4.52 (q, J = 7.3Hz, 1H), 4.18-4.10 (m, 2H), 1.59 (d, J = 7.3Hz, 3H), 1.23-1.18 (m, 3H). LCMS(ESI):m / z:462.74[M+1].

[0398] Compound 14F:

[0399]

[0400] To a solution of compound 14E (370 mg, 799.58 μmol) in dioxane (5 ml) were added compound tetrahydropyranamine (202 mg, 2.00 mmol) and DIEA (207 mg, 1.60 mmol, 278.98 μl). The reaction solution was stirred at 90°C for 4 hours. The reaction solution was diluted with water (10 ml) and extracted with ethyl acetate (10 ml x 3 times). The combined organic phases were dried and concentrated under reduced pressure to give a crude product 14F which was directly used in the next step. 1 H NMR (CDCl3, 400MHz) δ = 8.37 (s, 1H), 8.13 (d, J = 1.0Hz, 1H), 7.94 (dd, J = 1.6, 8.0Hz, 1H), 7.53-7.38 (m, 1H), 4.52 (q, J = 7.3Hz, 1H), 4.2 0-4.10(m,2H),3.96-3.89(m,2H),3.52-3.42(m,2H),2.01-1.91(m,2H),1.59(d,J=7.4Hz,3H),1.51-1.47(m,3H),1.22-1.20(m,3H). LCMS(ESI):m / z:527.43[M+1].

[0401] Compound 14G:

[0402]

[0403] To a solution of compound 14F (370 mg, 701.52 μmol) in tetrahydrofuran (2 ml) was added water and a solution of lithium hydroxide (59 mg, 1.4 mmol) in water (2 ml), and the reaction solution was stirred at 20°C for 0.5 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure, the pH of the reaction solution was adjusted to 2 with a 2-mol aqueous hydrochloric acid solution, and then extracted with ethyl acetate (10 ml x 3 times). The combined organic phase was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude compound 14G. 1 H NMR (400MHz, DMSO-d6) δ = 8.54 (s, 1H), 8.08 (s, 1H), 8.04-7.95 (m, 1H), 7.76 (d, J = 8.0Hz, 1H), 4.41 (q, J = 7.3Hz, 1H), 3.97-3.90 (m, 1H), 3.86 (br d,J=10.8Hz,2H),3.42-3.38(m,2H),1.84(brd,J=10.1Hz,2H),1.56-1.47(m,5H).

[0404] Compound 14:

[0405]

[0406] HATU (376 mg, 988.88 μmol) and DIEA (170 mg, 1.32 mmol, 229.11 μl) were added to a solution of compound 14G (330 mg, 660.83 μmol) and (2S)-2-amino-2-(3-fluoro-5-methoxy-phenyl)ethanol (176 mg, 794.02 μmol) in dichloromethane (5 ml), and the reaction solution was stirred at 20°C for 1 hour. The reaction solution was diluted with water (10 ml) and extracted with dichloromethane (20 ml × 3 times). The organic phase was washed with brine (20 ml × 1 time), dried, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was separated by high performance liquid chromatography (trifluoroacetic acid system) to obtain the free base of compound 14. 1 H NMR (400MHz, CD3OD) δ = 8.48 (s, 1H), 8.17 (d, J = 1.0Hz, 1H), 8.10 (dd, J = 1.6, 8.1Hz, 1H) ,7.70(d,J=8.4Hz,1H),6.77(s,1H),6.73-6.67(m,1H),6.61(td,J=2.3,10.8Hz,1H),4 .98-4.91(m,1H),4.61(q,J=7.0Hz,1H),4.10-4.02(m,1H),4.01-3.94(m,2H),3.83-3. 79(m,3H),3.78-3.68(m,2H),3.59-3.49(m,2H),2.04-1.96(m,2H),1.68-1.56(m,5H). LCMS(ESI):m / z:666.10[M+1].

[0407] Activity test

[0408] Experimental Example 1: Calu-6 (Kras Q61K ) Antiproliferative activity assay

[0409] Experimental Materials:

[0410] name Brand No. Calu-6 cells ATCC-HTB-56 RPMI1640 medium Cibco-22400-089 Fetal bovine serum Cellmax-BL100-02 L-Glutamine Gibco-35050-061 DPBS Corning-21-031-CVR Trypsin Gibco-25200-072 Double antibiotics (penicillin, streptomycin) Merck-TMS-AB2-C CellTiterGlo Promega-g7573 Cell Plate Greiner-781091 Echo Shallow Orifice Plate Labcyte-LP-0200

[0411] Experimental steps:

[0412] Cell seeding:

[0413] (1) Cell culture medium: 88% RPMI-1640, 10% fetal bovine serum, 1% L-glutamine and 1% penicillin-streptomycin;

[0414] (2) Preheat the culture medium, trypsin, and DPBS in a 37°C water bath;

[0415] (3) Remove the original culture medium in the cell culture flask and wash once with 6 ml of PBS;

[0416] (4) Add 3.5 ml of pancreatin to the cell culture flask, shake gently to allow the pancreatin to fully contact the cells, then remove the pancreatin, and place the culture flask in a 37°C incubator containing 5% CO2 for about 1 minute;

[0417] (5) Resuspend the cells in 10 ml of cell culture medium and take out about 0.6 ml of cell suspension for counting (ViCell XR);

[0418] (6) Dilute the cell suspension with culture medium to the cell density required for plating 2.5×10 4 cells per ml;

[0419] (7) Add 100 μl of PBS to each well around the cell plate, add 40 μl of cell suspension to other wells, and culture in a 37°C incubator containing 5% CO2 overnight.

[0420] (8) Take the required amount of cells and culture medium and continue culturing in a new T75 culture flask.

[0421] Dosing:

[0422] (1) Prepare the test compound into a 10 mM solution using DMSO;

[0423] (2) 9 μl of compound was added to an Echo shallow well plate (Labcyte, #LP-0200) and the shallow well plate was centrifuged at 1000 rpm for 10 seconds;

[0424] (3) Aspirate the DPBS outside the cell plate;

[0425] (4) Use Echo to perform gradient dilution and drug addition of the compounds. Dilute each compound into 10 concentration gradients and add 100 nanoliters to the 384 cell plate. Then, place the cell plate back into the incubator and culture for three days.

[0426] (5) Add 100 μl of DPBS to the periphery of the cell plate.

[0427] Read the plate and analyze the data:

[0428] (1) Add CTG and read the plate: Add 20 μl of CellTiterGlo to each well of the cell plate, shake in the dark for 10 min, and read the plate on Envision.

[0429] Experimental results:

[0430]

[0431] Experimental conclusion: The compound of the present invention has certain anti-proliferation activity on Calu-6 cells.

[0432] Experimental Example 2: HCT116 (Kras G13D ) Antiproliferative activity experiment Experimental materials:

[0433] 1) Experimental reagents and consumables:

[0434] name Brand No. Mc'Coy 5A Medium BI-01-075-1ACS Fetal bovine serum Biosera-FB-1058 / 500 0.25% Trypsin Source-S310KJ Double antibiotics (penicillin, streptomycin) Procell-PB180120 CellTiter Glo Promega-G7573 Cell Plate Corning-3610

[0435] 2) Experimental instruments:

[0436] name Brand No. Cell counting chamber Seeking Refinement Victor Nivo PerkinElmer

[0437] Experimental steps:

[0438] Cell seeding:

[0439] (1) Cell culture medium: 89% Mc'Coy 5A, 10% fetal bovine serum and 1% penicillin-streptomycin;

[0440] (2) Culture medium and trypsin were preheated in a 37°C water bath;

[0441] (3) Remove the culture medium from the cell culture flask and wash once with 1 ml of trypsin;

[0442] (4) Add 1 ml of pancreatin to the cell culture flask, shake gently to allow the pancreatin to fully contact the cells, then remove the pancreatin. Place the culture flask in a 37°C incubator containing 5% CO2 for about 1 min.

[0443] (5) Resuspend the cells in 2 ml of cell culture medium and take out about 0.01 ml of cell suspension for counting;

[0444] (6) Dilute the cell suspension with culture medium to the cell density required for plating 2.5×10 4 cells per ml;

[0445] (7) Add 100 μl of culture medium to each well around the cell plate, add 80 μl of cell suspension to the other wells, and place

[0446] The cells were cultured overnight in a 37°C incubator with 5% CO2.

[0447] (8) Take the required amount of cells and culture medium and continue culturing in a new T75 culture flask.

[0448] Dosing:

[0449] (1) Prepare the test compound into a 10 mmol solution using DMSO:

[0450] (2) The compound was diluted 3-fold in 9 concentration gradients, i.e., from 6 mmol to 2.7 μmol. A double-dip assay was set up, 78 μl of culture medium was added to the middle plate, and then 2 μl of the gradient dilution compound per well was transferred to the middle plate according to the corresponding position. After mixing, 20 μl of each well was transferred to the cell plate. The final concentration of the compound transferred to the cell plate was 30 μmol to 13.7 nanomolar. The cell plate was placed in a CO2 incubator and incubated for another 3 days.

[0451] 1) Read the plate and analyze the data:

[0452] (1) Add CTG and read the plate: Add 20 μL of CellTiter Glo to each well of the cell plate and shake in the dark for 10 min;

[0453] (2) Read the plate on a Victor Nivo;

[0454] Data Analysis:

[0455] The raw data were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by four-parameter curve fitting (obtained by "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism).

[0456] Experimental results:

[0457]

[0458] Experimental conclusion: The compounds of the present invention have good anti-proliferation activity against HCT116 cells.

[0459] Experimental Example 3: A375 (BRAF V600E ) Antiproliferative activity assay

[0460] Experimental Materials:

[0461] Cell line A375 (purchased from Pronocell), DMEM medium, penicillin / streptomycin antibiotics were purchased from Vicente, fetal bovine serum was purchased from Biosera, and CellTiter-glo (cell viability chemiluminescence detection reagent) reagent was purchased from Promega.

[0462] Experimental steps:

[0463] A375 cells were seeded in a white 96-well plate, with 80 micromolar cell suspension per well, containing 2000 A375 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture. The compound to be tested was diluted 3 times to the 9th concentration, i.e., from 6 millimolar to 0.91 micromolar, with a double-well experiment. 78 micromolar culture medium was added to the middle plate, and then 2 micromolar per well of the gradient dilution compound was transferred to the middle plate according to the corresponding position, and 20 micromolar per well was transferred to the cell plate after mixing. The concentration range of the compound transferred to the cell plate was 30 micromolar to 4.57 nanomolar. The cell plate was placed in a carbon dioxide incubator for 5 days. Prepare another cell plate, and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis. Add 25 micromolar cell viability chemiluminescent detection reagent to each well of this cell plate, and incubate at room temperature for 10 minutes to stabilize the luminescent signal. Reading was performed using a multi-label analyzer. After the cell plate with the compound added was incubated, 25 micromoles of cell viability chemiluminescent detection reagent was added to each well of the cell plate, and incubated at room temperature for 10 minutes to stabilize the luminescent signal. The reading was performed using a multi-label analyzer.

[0464] Data Analysis:

[0465] The raw data were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by four-parameter curve fitting (obtained in "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism). Experimental results:

[0466]

[0467] Experimental conclusion: The compounds of the present invention have good anti-proliferation activity against A375 cells.

[0468] Experimental Example 4: Colo205 (BRAF V600E ) Antiproliferative activity assay

[0469] Experimental Materials:

[0470] Cell line COLO205 (purchased from Pronocell), RPMI1640 culture medium, penicillin / streptomycin antibiotics were purchased from Vicente, fetal bovine serum was purchased from Biosera, and CellTiter-glo (cell viability chemiluminescence detection reagent) reagent was purchased from Promega.

[0471] Experimental methods:

[0472] COLO205 cell anti-proliferation experiment:

[0473] COLO205 cells were seeded in a white 96-well plate, with 80 micromolar cell suspension per well, containing 3000 COLO205 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture. The compound to be tested was diluted 3 times to the 9th concentration, i.e., from 200 micromolar to 0.03 micromolar, using a lance, and a double-well experiment was set up. 78 micromolar culture medium was added to the middle plate, and then 2 micromolar gradient dilution compounds per well were transferred to the middle plate according to the corresponding position, and 20 micromolar per well were transferred to the cell plate after mixing. The concentration range of the compound transferred to the cell plate was 1 micromolar to 0.15 nanomolar. The cell plate was placed in a carbon dioxide incubator for 3 days. Prepare another cell plate, and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis. Add 25 micromolar cell viability chemiluminescent detection reagent to each well of this cell plate, and incubate at room temperature for 10 minutes to stabilize the luminescent signal. Reading was performed using a multi-label analyzer. After the cell plate with the compound added was incubated, 25 micromoles of cell viability chemiluminescent detection reagent was added to each well of the cell plate, and incubated at room temperature for 10 minutes to stabilize the luminescent signal. The reading was performed using a multi-label analyzer.

[0474] Data Analysis:

[0475] The raw data were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by four-parameter curve fitting (obtained in "log(inhibitor)vs.response--Variable slope" mode in GraphPad Prism). Experimental results:

[0476]

[0477] Experimental conclusion: The compound of the present invention has good anti-proliferation activity against Colo205 cells.

[0478] Experimental Example 5: Calu-6 (Kras Q61K )ERK phosphorylation inhibition assay

[0479] Experimental Materials:

[0480] Reagents and consumables:

[0481]

[0482] Main instruments:

[0483] instrument Manufacturer model Biological Safety Cabinets AIRTECH BSC-1304IIA2 Carbon dioxide incubator Thermo 311 Cell Counter BECKMAN Vi-cellXR ELISA reader PerkinElmer Envision Centrifuge Eppendorf Centrifuge 5810R

[0484] Compound information (other compounds are derived from the compound master solution prepared by L1000)

[0485] Cell Information:

[0486] Cell Name Culture conditions source Part Number Calu6 1640+10%FBS ATCC ATCC-HTB-56

[0487] Experimental steps and methods:

[0488] 1) Resuscitate cells and culture to the logarithmic growth phase, digest with trypsin, seed 96-well plates with 30,000 cells per well, and incubate in an incubator overnight.

[0489] 2) Add 1 μl of the gradient compound dissolved in DMSO to the 96-well plate and return it to the incubator for incubation for 1 hour.

[0490] 3) Take out the cell plate, pour off the culture medium, and add 50 μl of cell lysis buffer (containing 1% blocking peptide) to each well.

[0491] 4) Mix the lysate plate at 500 rpm and incubate at room temperature for 30 minutes.

[0492] 5) Prepare the antibody mixture in the kit at a ratio of 1:1:38.

[0493] 6) Transfer 16 μl of cell lysate to each well of the HTRF plate, and then add 4 μl of the prepared antibody mixture.

[0494] 7) After overnight incubation, the plate was read using Envision, and the fitting curve was obtained according to the ratio (ratio of Ex665 / Ex615 fluorescence intensity) and the four-parameter fitting formula of Graphpad was Y=Bottom+(Top-Bottom) / (1+10^((Log EC 50 -X)*HillSlope)) to calculate EC 50 .

[0495] Experimental results:

[0496]

[0497] Experimental conclusion: The compounds of the present invention have good Calu-6 ERK phosphorylation inhibitory activity.

[0498] Experimental Example 6 Pharmacokinetic study of single intravenous and oral administration in mice and dogs

[0499] Purpose:

[0500] This experiment was designed to study the pharmacokinetics of the test compound in mice and dogs after a single oral administration.

[0501] Sample collection and preparation:

[0502] After intravenous or oral administration, blood samples were collected and the actual blood collection time was recorded. After blood sample collection, it was immediately transferred to a labeled centrifuge tube containing K2-EDTA, and then centrifuged to obtain plasma. The plasma was transferred to a pre-cooled centrifuge tube, quickly frozen in dry ice, and stored in an ultra-low temperature freezer at -70±10℃ until LC-MS / MS analysis.

[0503] Pharmacokinetic data analysis:

[0504] The pharmacokinetic software was used to process the plasma drug concentration data of the compound using a non-compartmental model. max ) and peak time (T max ) and the quantifiable end time, which can be directly obtained from the plasma drug concentration-time graph. The following pharmacokinetic parameters were calculated using the log-linear trapezoidal method: half-life (T 1 / 2 ), apparent volume of distribution (V dss ) and clearance (Cl), the area under the time-plasma concentration curve from point 0 to the terminal time point (AUC 0-last ).

[0505] 1. Pharmacokinetic data of single intravenous and oral administration in mice

[0506] Pharmacokinetic parameters of the compounds of the present invention after single intravenous administration to mice

[0507]

[0508] Pharmacokinetic parameters of the compounds of the present invention after single oral administration to mice

[0509]

[0510] Experimental conclusion: The compound of the present invention is well absorbed orally in mice and has a high exposure amount.

[0511] 2. Pharmacokinetic data of single intravenous and oral administration in dogs

[0512] Pharmacokinetic parameters of the compounds of the present invention after single intravenous and oral administration in dogs

[0513]

[0514] Experimental conclusion: The compound of the present invention has good oral absorption in dogs, low clearance rate, high exposure, and good bioavailability.

[0515] Experimental Example 7: In vivo pharmacodynamics experiment of human lung cancer Calu-6 cell subcutaneous xenograft tumor BALB / c nude mouse model Experimental materials:

[0516] 1.1 Experimental animals and breeding environment

[0517] 1.1.1 Experimental animals

[0518] Species: Mouse

[0519] Strain: BALB / c nude mice

[0520] Arrival age: 6-8 weeks old

[0521] Gender: Female

[0522] 1.1.2 Rearing environment

[0523] Animals were kept in SPF animal rooms in IVC (independent ventilation system, constant temperature and humidity) cages (3-5 per cage)

[0524] Temperature: 20-26℃

[0525] Humidity: 40-70%

[0526] 1.2 Tumor tissue or cell information

[0527] Cells: Human lung cancer Calu-6 cells were cultured in vitro, with 0.2 Units / ml bovine insulin and 10% fetal bovine serum added to EMEM medium, and cultured in a 37°C 5% CO2 incubator. The cells were routinely digested and subcultured with trypsin-EDTA twice a week. When the cell saturation reached 80%-90% and the number reached the required level, the cells were harvested, counted, and inoculated.

[0528] 1.3 Other reagent information

[0529] name Manufacturer Part Number Storage conditions Fetal bovine serum Hyclone SV30087.03 -20℃ Trypsin Gibco 25200-072 -20℃ EMEM medium ATCC ATCC30-2003 2-8℃

[0530] 1.4 Instrument information

[0531] name Manufacturer model Carbon dioxide incubator Thermo Fisher Heracell240i Low temperature high speed centrifuge Eppendorf 5810R Analytical Balance Sartorius SECURA225D-1CN Ordinary balance Changzhou Tianzhiping Instrument Equipment Co., Ltd. EL-2KJ Digital Vernier Caliper Mitutoyo 0~150mm

[0532] Experimental methods and steps:

[0533] 2.1 Tumor cell inoculation

[0534] Cell inoculation: 0.2 ml of Calu-6 cells (1:1 ratio with Matrigel) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached 173 mm 3 Start group dosing.

[0535] 2.2 Grouping

[0536] Table 1 Experimental animal groups and dosing regimen

[0537]

[0538] Note: 1: Number of mice in each group; 2: Dosing volume parameters: 10 μL / g based on the weight of the mouse. If the body weight drops by more than 15%, stop the drug until the body weight returns to within 10%; 3: 0.5% MC (methylcellulose).

[0539] 2.3 Tumor measurements and experimental parameters

[0540] Tumor diameter was measured with a vernier caliper twice a week. The formula for calculating tumor volume was: V = 0.5a × b 2 , a and b represent the long and short diameters of the tumor, respectively.

[0541] The anti-tumor efficacy of the compound was evaluated by TGI (%) or relative tumor proliferation rate T / C (%). Relative tumor proliferation rate T / C (%) = T RTV / C RTV ×100%(T RTV : Average RTV value of the treatment group; C RTV : The average RTV value of the negative control group). The relative tumor volume (RTV) was calculated based on the results of tumor measurement. The calculation formula is RTV = V t / V0, where V0 is the tumor volume measured at the time of group administration (i.e., D0), V t is the tumor volume at a certain measurement, T RTV With C RTV Get data from the same day.

[0542] TGI (%) reflects the tumor growth inhibition rate. TGI (%) = [1-(average tumor volume at the end of drug administration in a treatment group - average tumor volume at the beginning of drug administration in the treatment group) / (average tumor volume at the end of treatment in the solvent control group - average tumor volume at the beginning of treatment in the solvent control group)] × 100%.

[0543] 2.5 Statistical analysis

[0544] Statistical analysis was performed using SPSS software based on the RTV data at the end of the experiment. One-way ANOVA was used to analyze the inter-group comparisons. If the variance was not homogeneous (the F value was significantly different), the Games-Howell method was used for testing. p<0.05 was considered to be significantly different.

[0545] 3. Experimental results

[0546] 3.1 Inhibitory effect of the test substance on the growth of subcutaneous transplanted tumors of human lung cancer in nude mice

[0547] This experiment evaluated the efficacy of the test substance in a human lung cancer xenograft tumor model, with the solvent control group as a reference. The tumor volume of each group at different time points was as follows Figure 1As shown in the figure, the TGI of compound 1, compound 3, compound 9 and compound 11 in the administration group was 107%, 110%, 108% and 109%, respectively, which had a significant tumor inhibition effect (P<0.01).

[0548] 3.2 Weight changes

[0549] The weight and condition of mice were not significantly abnormal. Figure 2 .

[0550] Experimental conclusion:

[0551] The compound of the present invention has a significant inhibitory effect on the growth of tumor-bearing mice in a subcutaneous xenograft tumor model of human lung cancer Calu-6 cells.

Claims

1. A compound represented by formula (II) or a pharmaceutically acceptable salt thereof, in, T1 is CH or N; n is 1 or 2; R1 and R2 are each independently H, D, F, Cl or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br and I; R3 and R4 are each independently H or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH; R5 and R6 are each independently H or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3; R7 is phenyl or pyridyl, wherein the phenyl and pyridyl groups are optionally substituted by 1, 2, 3 or 4 R a replaced by; R8 is H, F, Cl or Br; R9 is tetrahydro-2H-pyranyl, wherein the tetrahydro-2H-pyranyl is optionally substituted with 1, 2, 3 or 4 R b replaced by; Each R a are independently F, Cl, Br, I, C 1-3 Alkyl, C 1-3 Alkoxy, NH-C 1-3 Alkyl or N-(C 1-3 alkyl)2, wherein the C 1-3 Alkyl, C 1-3 Alkoxy, -NH-C 1-3 Alkyl and -N-(C 1-3 alkyl)2 are each independently optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH; Each R b are independently F, Cl, Br, I, D or C 1-3 Alkyl, wherein the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH.

2. The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound has a structure represented by formula (I-1) or (II-1): in, R1, R2, R3, R4, R5, R6, R7, R8 and R9 are as defined in claim 1.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein each R b are independently F, Cl, Br, I, D or -CH3.

4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein R9 is It is stated Optional 1, 2, 3 or 4 R b replaced.

5. The compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein R9 is 6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure represented by formula (III-1): in, m is 0, 1, 2, 3 or 4; T1, R b , R1, R2, R3, R4, R5, R6, R7 and R8 are as defined in claim 1.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure represented by formula (I-3): in, R1, R2, R3, R4, R5, R6, R7 and R8 are as defined in claim 6.

8. The compound according to any one of claims 1, 2, 6 or 7, or a pharmaceutically acceptable salt thereof, wherein R1 and R2 are each independently H, D, F, Cl or -CH3.

9. A compound according to any one of claims 1, 2, 6 or 7, or a pharmaceutically acceptable salt thereof, wherein R3 and R4 are each independently H or -CH3, wherein the -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH.

10. The compound according to claim 9 or a pharmaceutically acceptable salt thereof, wherein R3 and R4 are each independently H, -CH3 or 11. A compound according to any one of claims 1, 2, 6 or 7, or a pharmaceutically acceptable salt thereof, wherein R5 and R6 are each independently H or -CH3, wherein the -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3.

12. The compound according to claim 11 or a pharmaceutically acceptable salt thereof, wherein R5 and R6 are each independently H, -CH3 or 13. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure represented by formula (III-3): in, m is 0, 1, 2, 3 or 4; T1, R b , R7 and R8 are as defined in claim 1; R3 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH; R5 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3.

14. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure represented by formula (I-5): in, R7 and R8 are as defined in claim 1; R3 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I and -OH; R5 is -CH3, wherein said -CH3 is optionally substituted with 1, 2 or 3 substituents independently selected from F, Cl, Br, I, -OH and -OCH3.

15. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein each R a are independently F, Cl, Br, I, -CH3, -OCH3, -NH-CH3 or 16. A compound according to any one of claims 1, 2, 6, 7, 12 or 13 or a pharmaceutically acceptable salt thereof, wherein R7 is 17. The compound according to claim 16 or a pharmaceutically acceptable salt thereof, wherein R7 is 18. A compound of the following formula or a pharmaceutically acceptable salt thereof:

19. A compound of the following formula or a pharmaceutically acceptable salt thereof:

20. Use of the compound according to any one of claims 1 to 19 or a pharmaceutically acceptable salt thereof in the preparation of ERK1 / 2 inhibitor drugs.

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