A pyrimido-heterocyclic compound, a pharmaceutical composition comprising the same, and use thereof

Abstract

The present application relates to a kind of pyrimidine and heterocyclic compound, the pharmaceutical composition comprising it and its purposes.The compound is the compound shown in formula I or its pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, metabolite, prodrug, wherein, the R1-R4 And X, Y, Y1, Z and L group are as defined in the specification.The compound described in the present application can be used to prepare the drug for treating and / or preventing cancer.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology and relates to a pyrimidine heterocyclic compound, a pharmaceutical composition containing the same, and its uses. Background Technology

[0002] The Ras gene is an important proto-oncogene, named after its discovery in rat sarcoma virus. There are three characteristic genes in the ras gene family associated with human tumors: H-ras, K-ras, and N-ras, located on chromosomes 11, 12, and 1, respectively. The first two are transformation genes of rat sarcoma virus, while the latter was isolated from human neuroblastoma.

[0003] The Ras protein, encoded by the Ras gene, has a relative molecular mass of 2.1 kDa and is located on the inner side of the cell membrane. It consists of 188 or 189 amino acids. The Ras protein exists in two conformations: an active GTP-binding conformation and an inactive GDP-binding conformation. These two conformations can interconvert. By switching between these two active states, the Ras protein regulates multiple signaling pathways, including RAF-MEK-ERK and PI3K / Akt / mTOR, thereby regulating cell growth, survival, migration, and differentiation.

[0004] RAS point mutations occur at codons 12, 13, and 61. Following RAS gene mutations, the exchange frequency between Ras and GDP / GTP is accelerated, allowing Ras to bind to GDP for extended periods. This results in persistently activated RAS signaling, leading to uncontrolled cell proliferation. Therefore, overactivated Ras signaling can ultimately cause cancer. RAS gene mutations are found in approximately one-third of human malignancies. The most common oncogenic mutation in the Ras family is the KRAS mutation (85%), while oncogenic mutations are less common in members of the NRAS (12%) and HRAS (3%) families.

[0005] KRAS mutations are common in a variety of cancers, including pancreatic cancer (95%), colorectal cancer (45%), and lung cancer (25%).

[0006] KRAS mutations include G12C, G12D, and G12V. G12D mutations are prevalent in pancreatic cancer (25%), colorectal cancer (13.3%), non-small cell lung cancer (4.1%), and small cell lung cancer (1.7%). While drug research on KRAS G12C mutations has made significant progress, with several KRAS G12C inhibitors, such as AMG-510 and MRTX-849, entering clinical trials and demonstrating good therapeutic efficacy and safety, there is still no effective treatment for tumors with KRAS G12D mutations, indicating a significant clinical need. Therefore, developing highly active inhibitors targeting KRAS G12D mutations has the potential to treat various cancers and holds a promising market prospect. Summary of the Invention

[0007] The technical problem to be solved by this invention is to provide a novel compound, its pharmaceutical composition, and its uses. The compound of this invention possesses KRAS G12D inhibitory activity, providing a new commercial option for KRAS G12D inhibitors.

[0008] The present invention solves the technical problem through the following technical solution.

[0009] According to a first aspect of the present invention, a compound of Formula I or a pharmaceutically acceptable salt thereof, stereoisomer, tautomer, hydrate, solvate, metabolite, or prodrug is provided:

[0010]

[0011] in,

[0012] X is selected from N or CR5;

[0013] Z is selected from N or CR5;

[0014] Y is selected from single bonds, C1-C3 alkyl groups, oxygen, sulfur, -CO-, or NR5; each of the alkyl groups is optionally substituted with at least one R5 molecule;

[0015] Y1 is selected from single bonds, C1-C3 alkyl groups, oxygen, sulfur, -CO- or NR5, wherein each alkyl group is optionally substituted with at least one R5;

[0016] L is selected from a single bond or a C1-C5 alkyl group, each of which is optionally substituted with at least one R7 molecule;

[0017] n is selected from 0, 1, 2, 3 or 4;

[0018] The R1 is selected from H, halogen, cyano, hydroxyl, -CO2R5, -CON(R5)2, C1-C3 alkyl, C5-C6 heteroaryl, and wherein the alkyl and heteroaryl groups are each optionally substituted by at least one R6.

[0019] R2 is selected from C1-C 12 Alkyl, C1-C 12 Heteroalkyl, C3-C8 cycloalkyl, C3-C8 heterocycloalkyl, C6-C 12 Bridged heterocycloalkyl, C6-C 12 Spirocycloalkyl or C6-C 12 And heterocyclic alkyl, wherein the alkyl, heteroalkyl, cycloalkyl, heterocyclic alkyl, bridged heterocyclic alkyl, spirocyclic alkyl and spirocyclic alkyl are each optionally substituted with at least one R7;

[0020] The R3 is selected from aryl or heteroaryl, wherein the aryl and heterocyclic groups are each optionally replaced by at least one R7;

[0021] The R4 is selected from H, halogen, C1-C3 alkyl, C3-C8 cycloalkyl, C1-C3 alkoxy or C1-C3 haloalkoxy;

[0022] The R5 is independently selected from hydrogen, OH, CN, NH2, halogen, C1-C3 alkyl, C3-C8 cycloalkyl, C1-C3 alkoxy or C1-C3 haloalkoxy;

[0023] The R6 is independently selected from hydrogen, OH, CN, C1-C6 alkyl, C1-C6 cycloalkyl, or C1-C6 alkoxy.

[0024] The R7 is independently selected from hydrogen, halogen, OH, CN, NH2, and C6-C. 10 Aryl, C5-C 10 The aryl group, C1-C8 alkyl group, C1-C8 heteroalkyl group, C1-C3 haloalkyl group, C1-C3 haloalkoxy group, C3-C8 cycloalkyl group, C3-C8 heterocycloalkyl group, C2-C4 alkenyl group, C2-C4 alkynyl group, -S-C1-C8 alkyl group, -O-C1-C8 alkyl group, -S-C1-C3 haloalkoxy group, -O-C1-C3 haloalkoxy group, -N(R5)2, or -CH2C(=O)N(R5)2, wherein the aryl group, heteroaryl group, alkyl group, heteroalkyl group, cycloalkyl group or heterocycloalkyl group is optionally substituted by at least one R8 group;

[0025] The R8 is independently selected from hydrogen, OH, CN, NH2, halogen, C1-C3 alkyl, C3-C8 cycloalkyl, C1-C3 alkoxy or C1-C3 haloalkoxy.

[0026] The term "hetero" in the above-mentioned heteroatom or heteroatomic group refers to heteroalkyl, heterocycloalkyl, bridged heterocycloalkyl, spirocycloalkyl, fused heterocycloalkyl, and heteroaryl groups, where "hetero" is independently selected from N, -O-, -S-, -C(=O)N(R5)-, -N(R5)-, -NH-, -C(=O)O-, -C(=O)-, -C(=S)-, -S(=O)-, -S(=O)2-, and -N(R5)C(=O)N(R5)-.

[0027] Preferably, it is a compound as shown in any of formulas II, III, IV or V.

[0028]

[0029] Secondly, the present invention provides the following specific compounds:

[0030]

[0031] Thirdly, the present invention provides the following specific compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, solvates, metabolites, and prodrugs.

[0032]

[0033]

[0034] Fourthly, in this invention, those skilled in the art can select the groups and substituents described in the general formula of the compound to provide a stable compound or its pharmaceutically acceptable salt, stereoisomer, tautomer, hydrate, solvate, metabolite, or prodrug, including but not limited to the compounds described in the embodiments of this invention.

[0035] Fifthly, the compounds provided by the present invention, or their pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, solvates, metabolites, prodrugs, or pharmaceutical compositions thereof, can be used to prepare KRAS G12D inhibitors for the treatment of diseases caused by KRAS G12D mutations.

[0036] In a sixth aspect, in some embodiments of the present invention, the use of the above-mentioned compounds provided by the present invention or their pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, solvates, metabolites, prodrugs, or pharmaceutical compositions thereof in the preparation of drugs for treating and / or preventing cancer, wherein the compounds described in the present invention can be used to treat and / or prevent cancer, wherein the cancers that can be used to treat and / or prevent include, but are not limited to, pancreatic cancer, colorectal cancer, and lung cancer.

[0037] Some solutions in this invention are derived from arbitrary combinations of the above-mentioned variables.

[0038] Technical effects:

[0039] This invention synthesizes a series of novel pyrimidine heterocyclic compounds. The compounds described in this invention can be used as drugs prepared as KRAS G12D inhibitors for the prevention and / or treatment of diseases with KRAS G12D mutations, and as drugs for the treatment and / or prevention of cancer, wherein the cancers treated and / or prevented include, but are not limited to, pancreatic cancer, colorectal cancer, and lung cancer.

[0040] Terms and Definitions

[0041] Unless otherwise stated, the terms used in the specification and claims shall have the following meanings.

[0042] "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 20 carbon atoms, such as straight-chain and branched groups with 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. In this document, "alkyl" can be a monovalent, divalent, or trivalent group. Non-limiting embodiments include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, and various branched isomers thereof. Non-limiting embodiments also include methylene, methine, ethylene, methine, propylene, methine, butylene, methine, and various branched isomers thereof. Alkyl groups may be optionally substituted or unsubstituted.

[0043] "Cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 12 ring atoms, for example, 3 to 12, 3 to 10, or 3 to 6 ring atoms, or 3, 4, 5, or 6-membered rings. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cyclohepttrienyl, cyclooctyl, etc. The cycloalkyl group may be optionally substituted or unsubstituted.

[0044] "Heterocyclic alkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, for example, 3 to 16, 3 to 12, 3 to 10, or 3 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O). mThe heteroatom (where m is 0, 1, or 2) excluding the -OO-, -OS-, or -SS- ring portion, with the remaining ring atoms being carbon. Preferably, it comprises 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, the heterocyclic alkyl ring comprises 3 to 10 ring atoms; most preferably, it is a 5-membered or 6-membered ring, of which 1 to 4 are heteroatoms; more preferably, 1 to 3 are heteroatoms; and most preferably, 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclic groups include spirocyclic, fused-ring, or bridged-ring heterocyclic groups.

[0045] "Spirocycloheterocyclic group" refers to a polycyclic group with 5 to 18 members, two or more ring structures, in which the monocyclic rings share an atom with each other, and contains one or more double bonds within the rings, but none of the rings has fully conjugated electrons. One or more ring atoms are selected from nitrogen, oxygen, or S(O). P (where p is selected from 0, 1 or 2) heteroatoms, and the remaining ring atoms are carbon. Preferably, it is 6 to 14-membered, more preferably 7 to 10-membered. Spirocyclic groups are classified into monospirocyclic groups, bispirocyclic groups or multispirocyclic groups according to the number of shared spiro atoms between rings, preferably monospirocyclic groups or bispirocyclic groups. More preferably, it is a 3-membered / 6-membered, 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered or 5-membered / 6-membered monospirocyclic groups. Wherein, "a-membered / b-membered monospirocyclic group" refers to a spirocyclic group in which the a-membered monocyclic ring and the b-membered monocyclic ring share one atom with each other. Non-limiting examples of "spirocyclic group" include, but are not limited to: diazaspiro[3.3]heptane.

[0046] "Fused heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system. One or more rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. One or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O)z (where z is an integer from 0 to 2), and the remaining ring atoms are carbon. Preferably, it is 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of constituent rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic group, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include: diazabicyclo[3.1.1]heptane.

[0047] "Bridged heterocyclic group" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two non-directly connected atoms. It may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. One or more ring atoms are heteroatoms selected from nitrogen, oxygen, or S(O)z (where z is an integer from 0 to 2), and the remaining ring atoms are carbon. Preferably, it is 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of constituent rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include: 6-azabicyclo[3.1.1]heptane.

[0048] The heterocyclic ring includes heterocyclic groups (including monocyclic, spirocyclic, fused heterocyclic and bridged heterocyclic) fused to an aryl, heteroaryl or cycloalkyl ring as described above, wherein the ring connected to the parent structure is a heterocyclic group, and non-limiting examples include 2,3-dihydrobenzofuran.

[0049] "Halogenated alkyl" or "halogenated alkoxy" means that the alkyl or alkoxy group is replaced by one or more identical or different halogen atoms. Preferred examples of alkyl or alkoxy groups include, but are not limited to, trifluoromethyl, trifluoroethyl, and trifluoromethoxy.

[0050] "Aryl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic system containing 6-14 ring atoms, wherein at least one ring system is aromatic, and each ring system comprises a ring of 3-7 atoms with one or more bonding sites connected to the rest of the molecule. Examples include, but are not limited to, phenyl, naphthyl, and anthracene. Preferably, the aryl group is a carbocyclic system with 6-10 or 6-7 ring atoms.

[0051] "Heteroaryl" refers to a monocyclic, bicyclic, or tricyclic system containing 5-14 ring atoms, wherein at least one ring system is aromatic, and at least one ring system contains one or more heteroatoms selected from nitrogen, oxygen, and sulfur, wherein each ring system comprises a ring of 5-7 atoms and has one or more bonding sites connected to the rest of the molecule. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic ring" or "heteroaromatic compound." Examples include, but are not limited to: furanyl, imidazolyl, 2-pyridyl, 3-pyridyl, thiazolyl, purinyl, and quinolinyl. Preferably, the heteroaryl group is a ring system with 5-10 ring atoms.

[0052] "Halogen" refers to fluorine, chlorine, bromine and iodine, with fluorine, chlorine and bromine being preferred.

[0053] "Optional" or "optionally" means that the event or circumstance described below may, but does not have to, occur. This description includes the possibility that the event or circumstance may or may not occur. For example, "optionally alkyl-substituted heterocyclic group" means that the alkyl group may, but does not have to, be present. This description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

[0054] "Substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents.

[0055] "Pharmaceutically acceptable salt" refers to the salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of 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 this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of 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, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; as well as salts of amino acids (such as arginine), and salts of organic acids such as glucuronic acid (see Berge et al., “Pharmaceutical Salts”, Journal of Pharmaceutical Science 66:1-19 (1977)). Certain specific compounds of the present invention contain basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt. Preferably, the salt is contacted with a base or acid in a conventional manner, and then the parent compound is separated, thereby regenerating the neutral form of the compound. The parent form of the compound differs from its various salt forms in certain physical properties, such as different solubilities in polar solvents.

[0056] "Pharmaceutical composition" means a mixture containing one or more compounds of Formula I described herein, or pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, solvates, metabolites, prodrugs, and other chemical components, as well as other components such as pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0057] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0058] In addition to the salt form, the compounds provided by this invention also exist in prodrug form. The prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to be converted into the compounds of this invention. Furthermore, the prodrugs can be converted into the compounds of this invention in the in vivo environment via chemical or biochemical methods.

[0059] Some compounds of this invention may exist in non-solventized or solvated forms, including hydrated forms. Generally, solvated and non-solventized forms are equivalent and both are included within the scope of this invention.

[0060] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- and (L)-enantiomers, racemic mixtures thereof, and other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0061] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0062] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0063] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0064] Unless otherwise stated, "(D)" or "(+)" indicates right-handed rotation, "(L)" or "(-)" indicates left-handed rotation, and "(DL)" or "(±)" indicates racemic rotation.

[0065] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by straight solid lines () and straight dashed lines (). The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and straight dashed key

[0066] The compounds of this invention can exist in specific forms. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. If tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomer can be achieved. For example, proton-tautomers (also called prototropic-tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence-tautomers include interconversions involving the rearrangement of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between the two tautomers, pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0067] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" 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%.

[0068] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0069] Optically active (R)- and (S)- isomers, as well as (D)- and (L)- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, 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 a 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 salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography with a chiral stationary phase, optionally combined with chemical derivatization (e.g., from amines to carbamates). The compounds of the present invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, compounds can be labeled with radioactive isotopes, such as tritium ( 3 H), Iodine-125 125 1) or C-14 ( 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0070] Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. Detailed Implementation

[0071] The preparation of the compounds shown in this invention or their pharmaceutically acceptable salts, stereoisomers, tautomers, hydrates, solvates, metabolites, and prodrugs can be accomplished by the exemplary methods described in the following examples and by relevant published literature used by those skilled in the art, but these examples are not intended to limit the scope of this invention.

[0072] The structure of the compounds described in this invention was determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). NMR measurements were performed using a Bruker AVANCE-400 or Varian Oxford-300 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS). Chemical shifts were expressed as 10⁻¹⁰. -6 (ppm) is given as the unit.

[0073] MS measurements were performed using an Agilent SQD (ESI) mass spectrometer (manufacturer: Agilent, model: 6110) or a Shimadzu SQD (ESI) mass spectrometer (manufacturer: Shimadzu, model: 2020).

[0074] HPLC determinations were performed using an Agilent 1200DAD high-performance liquid chromatograph (Sunfirc C18, 150X4.6mm, 5µm column) and a Waters 2695-2996 high-performance liquid chromatograph (Gimini C18, 150X4.6mm, 5µm column).

[0075] The silica gel plates used for thin-layer chromatography are Qingdao Ocean GF254 silica gel plates. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15mm-0.2mm, while the silica gel plates used for thin-layer chromatography separation and purification of products have a diameter of 0.4mm-0.5mm.

[0076] Column chromatography typically uses Qingdao Marine 200-300 mesh silica gel as a carrier.

[0077] The known starting materials of this invention can be synthesized using or according to methods known in the art, such as those from Accela ChemBio Inc. and Beijing Coupled Chemicals.

[0078] Unless otherwise specified in the examples, the reactions were carried out under an argon or nitrogen atmosphere.

[0079] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0080] A hydrogen atmosphere refers to a reaction flask connected to a hydrogen balloon with a volume of approximately 1 L. Hydrogenation reactions are typically carried out under vacuum, filled with hydrogen gas, and repeated three times.

[0081] Unless otherwise specified in the examples, the reaction temperature is room temperature, and the temperature range is 20℃-30℃.

[0082] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent systems used in the reactions were A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvents was adjusted according to the polarity of the compounds.

[0083] Unless otherwise specified in the examples, the preparative HPLC (formic acid) method used refers to the separation of the compound by chromatography in a formic acid system (phase A: H2O + 0.225% formic acid, phase B: acetonitrile).

[0084] The eluent system for column chromatography and the developing solvent system for thin-layer chromatography used to purify the compound include A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and small amounts of triethylamine and acidic or basic reagents can also be added for adjustment.

[0085] The present invention will be described in detail below with reference to examples, but this does not imply any adverse limitation on the invention. The compounds of the present invention can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic 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. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention. The following synthetic schemes describe the steps for preparing the compounds disclosed in the present invention. Unless otherwise stated, each substituent has the definition as described in the present invention.

[0086] Option A:

[0087]

[0088] Compound A1 undergoes high-temperature cyclization with urea to give A2, which then reacts with phosphorus oxychloride to give A3. Compound A3 reacts with the corresponding piperazine derivative under alkaline conditions to give A4. Compound A4 undergoes substitution with the corresponding alcohol, thiol, or amine under a suitable strong base, such as sodium tert-butoxide, to give A5. Compound A5 undergoes coupling reaction with a suitable borate ester, boric acid, or tin reagent to give A6. Compound A6 undergoes deprotection under acidic conditions to give the derivative of formula II.

[0089] Option B:

[0090]

[0091] Compound B1 can be reacted with urea at high temperature for ring closure or with sodium cyanide in an alkaline environment to give B2, which then reacts with phosphorus oxychloride to give B3. Compound B3 reacts with the corresponding piperazine derivative under alkaline conditions to give B4. Compound B4 reacts with the corresponding alcohol, thiol, or amine under a suitable strong base, such as sodium tert-butoxide, to give B5. Compound B5 undergoes a coupling reaction with a suitable borate ester, boric acid, or tin reagent to give B6. Compound A6 undergoes a deprotection reaction under acidic conditions to give the derivative of formula III.

[0092] Option C:

[0093]

[0094] Compound C1 reacts with thionyl chloride to give compound C2, which then reacts with S-methylisothiourea sulfate hydrogen under alkaline conditions to give compound C3. The base used can be sodium hydrogen, sodium hydroxide, or potassium hydroxide, etc. Compound C3 undergoes intramolecular cyclization under alkaline conditions, such as with sodium hydrogen hydrate or cesium carbonate, to give compound C4, which then reacts with phosphorus oxychloride to give C5. Compound C5 reacts with the corresponding piperazine derivative under alkaline conditions to give C6. Compound C6 undergoes oxidation with an oxidizing agent, such as m-chloroperoxybenzoic acid or hydrogen peroxide, to give compound C7, which then undergoes substitution under alkaline conditions, such as with sodium tert-butoxide, to give C8. Compound C8 undergoes coupling reaction with a suitable borate ester, boric acid, or tin reagent to give C9. Compound C9 undergoes deprotection under acidic conditions to give a derivative of formula IV.

[0095] Option D:

[0096]

[0097] Compound D1 reacts with phosphorus pentachloride and phosphorus oxychloride to give compound D2, which then reacts with S-methylisothiourea sulfate hydrogen under alkaline conditions to give compound D3. The base used can be sodium hydrogen, sodium hydroxide, or potassium hydroxide, etc. Compound D3 undergoes intramolecular cyclization under alkaline conditions, such as sodium hydrogen or cesium carbonate, to give compound D4, which then reacts with phosphorus oxychloride to give D5. Compound D5 reacts with the corresponding piperazine derivative under alkaline conditions to give D6. Compound D6 undergoes oxidation with an oxidizing agent, such as m-chloroperoxybenzoic acid or hydrogen peroxide, to give compound D7, which then undergoes substitution under alkaline conditions, such as sodium tert-butoxide, to give D8. Compound C8 undergoes coupling reaction with a suitable borate ester, boric acid, or tin reagent to give D9. Compound D9 undergoes deprotection under acidic conditions to give the derivative of formula V.

[0098] Example 1: Preparation of compound I-1

[0099]

[0100] Synthesis route:

[0101]

[0102] Preparation method:

[0103] Step 1: Synthesize compound 1B

[0104] Compound 1A (25 g, 119.06 mmol) was added to SOCl2 (75 mL), followed by 3 drops of DMF. The reaction mixture was then incubated at 85 °C for 1 h. After the reaction was complete as shown by TLC, the reaction mixture was evaporated to dryness. No further purification was required, and the mixture was directly added to the next step.

[0105] MS(ESI): m / z 228[M+1] + .

[0106] Step 2: Synthesize compound 1C

[0107] Sodium hydroxide (21 g, 525 mmol) was dissolved in water (460 mL), and then cooled to 0 °C. S-methylisothiourea hydrogen sulfate (60 g, 390.64 mmol) was slowly added in batches. After the addition was complete, the mixture was stirred at 0 °C for 30 minutes. Then, compound 1B (27.2 g, 119.06 mmol) obtained in the previous step was dissolved in methyl tert-butyl ether (348 mL), and this solution was slowly added dropwise to the reaction mixture at 0-5 °C. After the addition was complete, the temperature was slowly raised to room temperature and reacted for 1 h. TLC showed that the reaction was complete. The organic layer was separated, and the aqueous layer was further extracted with ethyl acetate (100 mL × 3). The organic layers were combined and washed with saturated sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to give product 1C (33.13 g, pale yellow solid, yield: 98.62%).

[0108] MS(ESI): m / z 282[M+1] + .

[0109] Step 3: Synthesize compound 1D

[0110] Compound 1C (33 g, 116.97 mmol) was dissolved in DMF (330 mL), and then cesium carbonate (38.28 g, 117.44 mmol) was added. After the addition was complete, the reaction solution was heated to 90 °C and stirred for 3 h. After the reaction was completed by TLC, the reaction solution was added to water (1.5 L). The aqueous phase was extracted with ethyl acetate (500 mL × 3), and the aqueous phase was separated. The pH of the obtained aqueous phase was adjusted to 6 with 6N hydrochloric acid, and a large amount of solid precipitated. The solid was filtered to obtain the solid, washed with water, and then dried under vacuum to obtain compound 1D (8 g, yellow solid, yield: 27.84%).

[0111] MS(ESI): m / z 246[M+1] + .

[0112] Step 4: Synthesize compound 1E

[0113] Phosphorus oxychloride (30 mL) was added to compound 1D (3 g, 12.21 mmol), and the reaction solution was heated to 100 °C. After TLC showed no reaction, the solution was evaporated to dryness. The residue was slowly added to a saturated NaHCO3 solution (150 mL). The aqueous phase was extracted with (50 mL × 3) solutions, and the organic phases were combined and dried over anhydrous sodium sulfate. The residue was then evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1 (v / v)) to give compound 1E (3.1 g, yellow solid, yield: 96.12%).

[0114] MS(ESI): m / z 264[M+1] + .

[0115] Step 5: Synthesize compound 1F

[0116] Compound 1E (9 g, 34.4 mmol) was added to acetonitrile (100 mL) at room temperature, followed by (S)-4-N-tert-butoxycarbonyl-2-methylpiperazine (9.6 g, 52 mmol) and DIEA (6.7 g, 52 mmol). The mixture was then refluxed for 3 hours. TLC showed that the reaction was complete. The reaction solution was concentrated under vacuum, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1 (V:V volume ratio)) to give compound 1F (9 g, pale yellow solid), yield: 63.3%.

[0117] MS m / z(ESI): 428[M+1] + .

[0118] Step 6: Synthesize compound 1H

[0119] Compound 1F (1.2 g, 2.80 mmol) and 1G (0.78 g, 3.1 mmol) were dissolved in dioxane (20 mL) and water (4 mL). Pd(dppf)₂Cl₂·DCM (0.11 g, 0.14 mmol) and potassium carbonate (0.78 g, 5.6 mmol) were added, and the reaction mixture was reacted at 100 °C for 12 hours. After the reaction was complete as shown by TLC, the reaction mixture was added to water (100 mL), and the aqueous phase was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and washed with saturated sodium chloride (100 mL × 3). The organic phase was separated and dried over anhydrous sodium sulfate. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 1H (1.2 g, brownish-yellow solid, yield 82%).

[0120] MS(ESI): m / z 520[M+1] + .

[0121] Step 7: Synthesize compound 1I

[0122] Compound 1H (0.12 g, 0.23 mmol) and sodium tert-butoxide (44 mg, 0.46 mmol) were dissolved in ethyl acetate (5 mL). m-CPBA (0.2 g, 0.92 mmol) was added in portions under ice bath conditions, and the reaction was carried out at 0 °C for 1 h. After the reaction was complete as indicated by TLC, the reaction mixture was poured into a saturated sodium thiosulfate aqueous solution (25 mL), extracted with ethyl acetate (15 mL × 3), and the organic phases were combined. The organic phase was washed with saturated sodium chloride (10 mL × 2), dried over anhydrous sodium sulfate, and evaporated to dryness. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1:1 (v / v)) to give compound 1I (0.1 g, brownish-yellow oil, yield 78%).

[0123] MS(ESI): m / z 552[M+1] + .

[0124] Step 8: Synthesize compound 1K

[0125] Compound 1I (0.1 g, 0.18 mmol) and 1J (29 mg, 0.18 mmol) were added to toluene (3 mL), and sodium tert-butoxide (35 mg, 0.36 mmol) was added in portions under ice bath conditions. The reaction was carried out under ice bath conditions for 1 h. After the reaction was completed by TLC, the reaction solution was added to a saturated sodium chloride aqueous solution (30 mL), and the aqueous phase was extracted with ethyl acetate (15 mL × 3). The organic phases were combined and washed with saturated sodium chloride (10 mL × 2). The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness. The residue was purified by prep-TLC (EA:MeOH = 8:1) to give compound 1K (70 mg, yellow solid, yield 61%).

[0126] MS(ESI): m / z 631[M+1] + .

[0127] Step 9: Synthesize compound I-1

[0128] Compound 1K (70 mg, 2.4 mmol) was added to acetonitrile (2 mL), followed by 8% ethyl hydrochloride (1 mL). The reaction mixture was allowed to react at room temperature for 1 h. After the reaction was complete as shown by TLC, the reaction mixture was concentrated under reduced pressure, and the residue was purified by slurrying with ethyl acetate to give compound I-1 (50 mg, yellow solid, 75% yield).

[0129] MS(ESI): m / z 531[M+1] + .

[0130] 1 H NMR (400MHz, DMSO-d6) δ11.90 (s, 1H), 10.13 (d, J = 12Hz, 1H), 9.77–9.60 (m, 1H), 8. 55(d,J=8Hz,1H),8.17-8.09(m,2H),7.78-7.72(m,3H),7.63-7.55(m,2H),5.69-5. 45(m,1H),5.08-5.06(m,1H),4.79-4.66(m,2H),4.46(d,J=16Hz,1H),3.81-3.69(m ,3H),3.38-3.29(m,6H),2.70-2.53(m,2H),2.37-2.08(m,4H),1.61(d,J=8Hz,3H).

[0131] Example 2: Preparation of compound I-2

[0132]

[0133] Synthesis route:

[0134]

[0135] Preparation method:

[0136] Step 1: Synthesize compound 2C

[0137] Compound 2A (3.1 g, 11.74 mmol) was added to DCM (31 mL), followed by compound 2B (2.49 g, 11.74 mmol). The mixture was then placed at 0 °C and DIEA (3.05 g, 23.66 mmol, 2.0 eq) was added dropwise. The reaction mixture was then allowed to warm naturally to room temperature for 1 hour. After the reaction was completed by TLC, the mixture was evaporated to dryness. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1 (v / v)) to give compound 2C (3.76 g yellow solid, yield: 72.81%).

[0138] MS(ESI): m / z 440[M+1] + .

[0139] Step 2: Synthesize compound 2E

[0140] Compound 2C (2 g, 4.55 mmol) and compound 2D (1.15 g, 4.55 mmol) were added to dioxane (20 mL), followed by potassium carbonate (1.26 g, 9.13 mmol), and then Pd(dppf)₂Cl₂DCM (372 mg, 0.45 mmol). The reaction mixture was then purged with nitrogen three times and reacted at 100 °C. After the reaction was complete as shown by TLC, the reaction mixture was added to a saturated sodium chloride aqueous solution (100 mL), and the aqueous phase was extracted with ethyl acetate (50 mL × 3). The organic layers were combined and dried over anhydrous sodium sulfate. The organic layer was then evaporated to dryness, and the residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1 (v / v)) to give compound 2E (1.91 g, brown solid, yield: 79.03%).

[0141] MS(ESI): m / z 532[M+1] + .

[0142] Step 3: Synthesize compound 2F

[0143] Compound 2E (600 mg, 1.13 mmol) was dissolved in DCM (6 mL), followed by the addition of m-CPBA (390 mg, 2.259 mmol q). The reaction was then carried out at 0 °C. After the reaction was complete as shown by TLC, saturated sodium thiosulfate aqueous solution (30 mL) was added. The aqueous phase was then extracted with dichloromethane (15 mL × 3). The organic phases were combined and washed with saturated NaCl solution (15 mL × 3). The organic layer was dried over anhydrous sodium sulfate and evaporated to dryness. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (v / v)) to give compound 2F (210 mg, pale white powder, yield: 33.01%).

[0144] MS(ESI): m / z 564[M+1] + .

[0145] Step 4: Synthesize compound 2H

[0146] Compound 2F (210 mg, 372.57 mmol) and 2G (118.63 mg, 745.15 mmol) were dissolved together in toluene (2 mL, 10 V) at 0 °C, followed by the addition of sodium tert-butoxide (35.84 mg, 373 mmol, 1 eq). The reaction mixture was then placed at 0 °C for 5 min. After the reaction was complete as shown by TLC, the reaction mixture was added to a saturated sodium chloride aqueous solution (20 mL). The aqueous phase was extracted with ethyl acetate (10 mL × 3). The organic phases were combined and washed with saturated NaCl solution (10 mL × 3). The organic layer was then dried over anhydrous sodium sulfate and evaporated to dryness. The residue was purified by pre-TLC (evolving solvent: EA / MeOH = 10 / 1) to give compound 2H (53 mg, yellow solid, yield: 22.13%).

[0147] MS(ESI): m / z 645[M+1] + .

[0148] Step 5: Synthesize compound I-2

[0149] Compound 2H (53 mg, 82.46 mmol, 1 eq) was dissolved in MeCN (2 mL), and then 8% ethyl acetate hydrochloride solution (1 mL) was added. The reaction solution was then naturally heated to room temperature and reacted for half an hour. After the reaction was completed by TLC, the reaction solution was evaporated to dryness, and then EA was added and evaporated to dryness. The solution was then slurried with EA and filtered to obtain compound I-2 (27 mg, yellow solid, 60.34%).

[0150] MS(ESI): m / z 545[M+1] +

[0151] 1H NMR (400MHz, DMSO-d6) δ11.78(s,1H),10.13(d,J=10.0Hz,1H),9.80(d,J=9.6Hz,1 H),8.61(d,J=9.8Hz,1H),8.18(n,J=7.7Hz,1H),8.10(n,J=8.1Hz,1H),7.98–7.51( n,5H),5.60(d,J=52.4Hz,1H),4.69(t,J=11.2Hz,3H),4.19(s,2H),4.07(d,J=13.8 Hz, 2H), 3.85 (s, 4H), 3.27 (d, J = 12.3Hz, 1H), 2.69–2.53 (n, 1H), 2.37–1.92 (n, 7H).

[0152] Example 3: Preparation of compound I-3

[0153]

[0154] Synthesis route:

[0155]

[0156] Preparation method:

[0157] Step 1: Synthesize compound 3B

[0158] Compound 3A (2 g, 12.3 mmol) was dissolved in THF (30 mL), and DMAP (0.15 g, 1.23 mmol) was added. Boc₂O (5.36 g, 24.5 mmol) was added dropwise with stirring at room temperature. The mixture was stirred at 70°C for 0.5 hours, followed by the addition of Boc₂O (5.36 g, 24.5 mmol). The reaction mixture was then stirred at 70°C for 1 hour. After the reaction was complete as shown by TLC, the reaction mixture was evaporated to dryness. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20:1 (v / v)) to give compound 3B (4.2 g, white solid, 94% yield).

[0159] 1H NMR (400MHz, CDCl3) δ7.12 (s, 2H), 1.51 (s, 18H).

[0160] Step 2: Synthesizing compound 3C

[0161] Compound 3B (4.5 g, 12.4 mmol) was dissolved in THF (60 mL), and LDA (2 M, 21.7 mL, 43.4 mmol) was added at -60 °C. The reaction was carried out at -60 °C for 0.5 h. After the reaction was completed by TLC, the reaction was quenched by pouring in saturated ammonium chloride aqueous solution (250 mL). The organic layer was separated, and the aqueous phase was extracted with ethyl acetate (3 × 150 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and evaporated to dryness. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 10:1 (v / v)) to give compound 3C (4 g, yellow solid, yield 89%).

[0162] 1H NMR (400MHz, CDCl3) δ8.99(s,1H),8.27(s,1H),1.58(s,9H)1.02(s,9H).

[0163] Step 3: Synthesizing Compound 3D

[0164] Compound 3C (4 g, 11.0 mmol) was added to ethyl acetate (25 mL), and the mixture was reacted with 8% ethyl acetate hydrochloride (25 mL) in an ice bath at 35 °C for 48 h. After the reaction was complete as shown by TLC, the mixture was evaporated to dryness, and the residue was purified by slurrying with MTBE. The residue was filtered to obtain a solid, which was then dried under vacuum to give compound 3D (2.6 g, white solid, 90% yield).

[0165] MS(ESI): m / z 263[M+1] + .

[0166] Step 4: Synthesize compound 3F

[0167] Compound 3D (2.5 g, 9.5 mmol) was added to THF (30 mL), and 3E (2.7 g, 14.3 mmol) was added dropwise under ice bath conditions. The reaction was carried out at room temperature for 1 h. After the reaction was completed by TLC, the mixture was evaporated to dryness, and the residue was purified by slurrying with petroleum ether. The residue was filtered to obtain a solid, which was then dried under vacuum to give compound 3F (4 g, white solid, 93% yield).

[0168] MS(ESI): m / z 450[M+1] + .

[0169] Step 5: Synthesize compound 3G

[0170] Compound 3F (4.5 g, 10 mmol) was dissolved in MeOH (40 mL), and 7 M NH3 methanol solution (5 mL) was added dropwise under ice bath. The reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC, the mixture was evaporated to dryness, and the residue was purified by slurrying with MTBE. The residue was filtered to obtain a solid, which was then dried under vacuum to give compound 3G (2.2 g, white solid, 95% yield).

[0171] MS(ESI): m / z 232[M+1] + .

[0172] Step 6: Synthesize compound 3H

[0173] Compound 3G (1 g, 4.3 mmol) was dissolved in MeOH (12 mL). 60% sodium hydroxide (0.69 g, 17.2 mmol) was added in portions under ice bath conditions. After the addition was complete, the reaction was allowed to proceed at room temperature for 30 minutes. TLC showed a large amount of reactant remaining. The temperature was then raised to 70 °C and reacted for 12 hours. After the reaction was complete under TLC, methanol was evaporated, and the residue was quenched in ice water. The pH was adjusted to 6 with 2 M hydrochloric acid under ice bath conditions, precipitating a white solid. The filtered solid was washed several times with ice-cold methanol and then dried under vacuum to give compound 3H (0.9 g, white solid, 92% yield).

[0174] MS(ESI): m / z 228[M+1] + .

[0175] Step 7: Synthesize compound 3I

[0176] Compound 3H (500 mg, 2.20 mmol) was added to POCl3 (6 mL), and DIEA (568 mg, 4.39 mmol) was added dropwise under ice bath conditions. The mixture was purged with nitrogen three times, and then the reaction solution was reacted at 110 °C for 2 hours. After the reaction was complete as shown by TLC, POCl3 was evaporated to dryness, and 30 mL of ice water was added to the residue. The pH was adjusted to 7 with saturated sodium bicarbonate aqueous solution, and the aqueous phase was extracted with ethyl acetate (3 × 15 mL). The organic phases were combined, washed with saturated sodium chloride aqueous solution, and dried with anhydrous sodium sulfate. The organic phase was evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5:1 (v / v)) to give compound 3I (520 mg, pale yellow solid, yield 89%).

[0177] MS(ESI): m / z 264[M+1] + .

[0178] Step 8: Synthesize compound 3K

[0179] Compound 3I (1.00 g, 3.78 mmol) was added to DCM (20 mL), followed by 3J (803 mg, 3.78 mmol). DIEA (977 mg, 7.56 mmol) was added dropwise at -40 °C and the reaction was allowed to proceed for 0.5 h. After the reaction was complete as shown by TLC, the mixture was extracted with water and ethyl acetate, washed with brine, and the organic phase was dried over anhydrous sodium sulfate. The residue was evaporated to dryness and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 3K (1 g, pale yellow oil, 60% yield).

[0180] MS(ESI): m / z 440[M+1] + .

[0181] Step 9: Synthesize compound 3M

[0182] Compound 3K (600 mg, 1.36 mmol) was added to tetrahydrofuran (10 mL), followed by 3 L (433 mg, 2.73 mmol). Sodium tert-butoxide (262 mg, 2.73 mmol) was added in portions at -40 °C, and the reaction was carried out for 0.5 h. After the reaction was completed by TLC, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: ethyl acetate / methanol = 20:1 (v / v)) to give compound 3M (430 mg, pale yellow oil, yield 56%).

[0183] MS(ESI): m / z 563[M+1] + .

[0184] Step 10: Synthesize compound 3O

[0185] Compounds 3M (0.25 g, 0.44 mmol) and 3M (0.22 g, 0.44 mmol) were dissolved in dioxane (6 mL) and water (2 mL). Pd(dppf)₂Cl₂·DCM (36 mg, 0.04 mmol) and cesium carbonate (0.29 g, 0.89 mmol) were added. The mixture was purged with nitrogen three times, and then the reaction solution was reacted at 102 °C for 18 hours. After the reaction was complete, water was added and the mixture was extracted with ethyl acetate, washed with brine, and the organic phase was dried over anhydrous sodium sulfate. The mixture was then evaporated to dryness, and the residue was purified by prep-TLC (ethyl acetate / methanol / ammonia = 20:1:0.001, v / v) to give compound 3O (50 mg, brown oil, 13% yield).

[0186] MS(ESI): m / z 895[M+1] + .

[0187] Step 11: Synthesize compound I-3

[0188] Compound 3O (50 mg, 0.06 mmol) was added to DMF (1.0 mL), followed by cesium fluoride (42 mg, 0.28 mmol). The reaction mixture was reacted at room temperature for 1 h. The mixture was extracted with ethyl acetate, washed with brine, and dried over anhydrous sodium sulfate. The solution was then evaporated to dryness to obtain a brown oil. This brown oil was dissolved in acetonitrile (2 mL), and 8% ethyl acetate hydrochloride (1 mL) was added under ice bath conditions. The reaction mixture was reacted at room temperature for 1 h. After the reaction was complete as indicated by TLC, the reaction solution was concentrated under reduced pressure. The residue was purified by prep-HPLC to give compound I-3 (10 mg, pale yellow solid, 30% yield).

[0189] MS(ESI): m / z 581[M+1] + .

[0190] 1H NMR (400MHz, DMSO-d6) δ11.34(s,1H),10.21(s,1H),7.87(d,J=7.8Hz,1H),7.54(d,J=6.4Hz,1H),7.44(t ,J=8.0Hz,1H),7.33(d,J=2.5Hz,1H),7.12(d,J=2.4Hz,1H),6.11(s,1H),5.35-5.20(m,1H),4.15-4.01(m ,2H),3.98-3.85(m,2H),3.82(s,1H),3.49-3.45(m,2H),3.27-3.19(m,2H),3.07(t,J=9.8Hz,2H),3.01( s,1H),2.83-8-2.76(m,1H),2.1-4-2.08(m,1H),2.05-1.94(m,3H),1.87-1.70(m,4H),1.65-1.55(s,2H).

[0191] Example 4: Preparation of compound I-4

[0192]

[0193] Synthesis route:

[0194]

[0195] Preparation method:

[0196] Step 1: Synthesize compound 4B

[0197] Compound 4A (25 g, 193 mmol) was dissolved in concentrated sulfuric acid (50 mL). A mixed solution of concentrated sulfuric acid (45.4 g, 463.17 mmol) and concentrated nitric acid (24.3 g, 386 mmol) was added under ice bath conditions. The reaction was then carried out under ice bath conditions for 1 h, followed by a 1.5 h reaction at room temperature. After the reaction was complete as indicated by TLC, the reaction solution was slowly added to ice water (500 mL), and the pH was adjusted to 7-8 with 15% NaOH aqueous solution. The organic phases were extracted with ethyl acetate (3 × 150 mL), combined, and dried over anhydrous sodium sulfate. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4B (6.0 g, yellow oil, 17% yield).

[0198] 1 H-NMR (400MHz, CDCl3) δ10.53 (s, 1H), 8.14 (d, J = 5.5Hz, 1H), 7.88 (d, J = 5.5Hz, 1H).

[0199] Step 2: Synthesize compound 4C

[0200] Compound 4B (6.0 g, 34.38 mmol) was dissolved in a mixed solvent of CH3CN (53 mL) and MeOH (7 mL). A solution of trimethylsilyl diazomethane-n-hexane (2 M, 34.4 mL, 68.8 mmol) was added dropwise under ice bath conditions, and the reaction was carried out at 0 °C for 0.5 h. After the reaction was completed as shown by TLC, the reaction was quenched in a saturated ammonium chloride aqueous solution (250 mL). The aqueous phase was extracted with ethyl acetate (3 × 150 mL). The organic phases were combined and dried over anhydrous sodium sulfate. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5:1 (v / v)) to give compound 4C (3.2 g, yellow oil, yield 49%).

[0201] MS(ESI): m / z 189[M+1] + .

[0202] Step 3: Synthesize compound 4D

[0203] Compound 4C (3.2 g, 16.97 mmol) was added to AcOH (32 mL), and iron powder (4.74 g, 84.85 mmol) was added with stirring at room temperature. The reaction was carried out at 40 °C for 1 h. After the reaction was completed by TLC, the mixture was evaporated to dryness, and the residue was diluted with ethyl acetate (150 mL). The residue was then added to a saturated sodium bicarbonate aqueous solution (500 mL), and the pH was adjusted to 7-8. The organic phase was separated, and the aqueous phase was extracted again with ethyl acetate (3 × 150 mL). The organic phases were combined and dried with anhydrous sodium sulfate. The mixture was evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4D (2.0 g, yellow oil, yield 74%).

[0204] MS(ESI): m / z 159[M+1] + .

[0205] Step 4: Synthesize compound 4E

[0206] Compound 4D (2.0 g, 12.53 mmol) was added to MeCN (20 mL), and NIS (4.23 g, 18.80 mmol) and p-toluenesulfonic acid monohydrate (240 mg, 1.25 mmol) were added in portions under ice bath conditions. The mixture was then heated to 70 °C and reacted for 16 h. After the reaction was complete as shown by TLC, the mixture was evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4E (3.2 g, yellow solid, 89% yield).

[0207] MS(ESI): m / z 285[M+1] + .

[0208] Step 5: Synthesize compound 4F

[0209] Compound 4E (3.0 g, 10.55 mmol) was added to MeOH (30 mL), followed by the dropwise addition of TEA (2.13 g, 21.1 mmol) and Pd(dppf)₂Cl₂·DCM (396 mg, 0.53 mmol). The reaction was carried out at 70 °C for 16 h under a carbon monoxide atmosphere (15 psi). After the reaction was completed by TLC, the reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4F (1.8 g, yellow solid, 78% yield).

[0210] MS(ESI): m / z 217[M+1] + .

[0211] Step 6: Synthesize compound 4G

[0212] Compound 4F (1.8 g, 8.31 mmol) was dissolved in THF (20 mL), and trichloroacetyl isocyanate (2.35 g, 12.46 mmol) was added dropwise under ice bath conditions. The reaction was allowed to proceed at room temperature for 0.5 h. After the reaction was complete as shown by TLC, the reaction mixture was evaporated to dryness and slurried with PE. The resulting white solid was added to MeOH (15 mL), and a 40% (w / w) methylamine alcohol solution (10 mL) was added dropwise under ice bath conditions. The reaction was allowed to proceed at 0 °C for 1 h, during which a large amount of white solid precipitated. After the reaction was complete as shown by TLC, the reaction mixture was evaporated to dryness, and the resulting solid was slurried with EA. The solid was filtered and dried under vacuum to give compound 4G (1.5 g, white solid, 80% yield).

[0213] MS(ESI): m / z 228[M+1] + .

[0214] Step 7: Synthesize compound 4H

[0215] Compound 4G (1.5 g, 6.6 mmol) was added to phosphorus oxychloride (20 mL), and DIEA (1.7 g, 13.2 mmol) was added dropwise under ice bath conditions. The reaction was carried out at 110 °C for 3 h. After the reaction was complete as shown by TLC, the mixture was evaporated to dryness. The residue was diluted with ethyl acetate (50 mL) and added to a saturated sodium bicarbonate aqueous solution (200 mL). The pH was adjusted to 7-8. The organic phase was separated, and the aqueous phase was extracted with ethyl acetate (3 × 50 mL). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was evaporated to dryness, and the residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5:1 (v / v)) to give compound 4H (920 mg, yellow solid, yield 52%).

[0216] MS(ESI): m / z 264[M+1] + .

[0217] Step 8: Synthesize compound 4J

[0218] Compound 4H (1 g, 3.78 mmol, WO2020146613) was dissolved in DCM (15 mL), followed by the addition of 4I (802 mg, 3.78 mmol). DIEA (977 mg, 7.56 mmol) was then added dropwise under ice bath conditions, and the reaction mixture was allowed to react at room temperature for 0.5 h. After the reaction was complete as shown by TLC, the reaction mixture was added to 50 mL of saturated sodium chloride aqueous solution, and the organic phase was separated. The water was then extracted with dichloromethane (3 × 15 mL). The organic phases were combined and dried over anhydrous sodium sulfate. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3:1 (v / v)) to give compound 4J (1.2 g, white solid, 72% yield).

[0219] MS(ESI): m / z 440[M+1]+.

[0220] Step 9: Synthesize compound 4L

[0221] Compound 4J (400 mg, 0.91 mmol) was dissolved in THF (6 mL), and 4K (144.62 mg, 0.91 mmol) was added. Sodium tert-butoxide (174 mg, 1.82 mmol) was added in portions at 0 °C, and the reaction mixture was brought to room temperature and reacted for 30 mins. After the reaction was complete as shown by TLC, the residue was purified by silica gel column chromatography (eluent: ethyl acetate / methanol = 20:1 (v / v)) to give compound 4L (320 mg, pale yellow solid, yield 62%).

[0222] MS(ESI): m / z 563[M+1] + .

[0223] Step 10: Synthesize compound 4N

[0224] Compounds 4L (0.25 g, 0.44 mmol) and 4M (0.22 g, 0.44 mmol) were dissolved in dioxane (6 mL) and water (2 mL). Pd(dppf)₂Cl₂·DCM (36 mg, 0.04 mmol) and cesium carbonate (0.29 g, 0.89 mmol) were added. The mixture was purged with nitrogen three times, and then the reaction solution was reacted at 102 °C for 18 hours. After the reaction was complete, water was added and the mixture was extracted with ethyl acetate, washed with brine, and the organic phase was dried over anhydrous sodium sulfate. The mixture was then evaporated to dryness, and the residue was purified by prep-TLC (ethyl acetate / methanol / ammonia = 20:1:0.001, v / v) to give compound 4N (50 mg, brown oil, 13% yield).

[0225] MS(ESI): m / z 851[M+1] + .

[0226] Step 11: Synthesize compound I-4

[0227] Compound 4N (51 mg, 0.06 mmol) was added to DMF (1.0 mL), followed by cesium fluoride (42 mg, 0.28 mmol). The reaction mixture was reacted at room temperature for 1 h. The mixture was extracted with ethyl acetate and washed with brine. The organic phase was dried over anhydrous sodium sulfate and evaporated to dryness to obtain a brown oil. This brown oil was then dissolved in acetonitrile (2 mL), and 8% ethyl acetate hydrochloride (1 mL) was added under ice bath conditions. The reaction mixture was reacted at room temperature for 1 h. After the reaction was complete as indicated by TLC, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC to give compound I-4 (13 mg, pale yellow solid, 33% yield).

[0228] MS(ESI): m / z 595[M+1] + .

[0229] 1 H-NMR (400MHz, DMSO-d6) δ11.22(s,1H),10.12(s,1H),9.47(d,J=8.9Hz,1H),9.23(s,1H),9.02(s,1H),7.8 6(dd,J=7.6,1.9Hz,1H),7.46–7.36(m,2H),9.29(d,J=4.0Hz,1H),7.04–7.01(m,1H),5.57(d,J=52.2Hz,1H) ,4.76–4.53(m,3H),4.50-4.45(m,1H),4.23-4.21(m,2H),3.93–3.83(m,3H),3.82(s,3H),3.7-3.75(m,1H), 3.49(s,1H),3.35-3.24(m,2H),2.58-2.53(m,2H),2.34-2.28(m,1H),2.26–2.12(m,2H),2.21-1.85(m,5H).

[0230] Example 5: Preparation of compound I-5

[0231]

[0232] Synthesis route:

[0233]

[0234] Step 1: Synthesize compound 5A

[0235] Compound 2C (2 g, 4.55 mmol, 1.0 eq) and compound 3N (2.2 g, 4.55 mmol, 1.0 eq) were added to dioxane (20 mL, 10 V), followed by potassium carbonate (1.26 g, 9.13 mmol, 2.0 eq), and then Pd(dppf)₂Cl₂DCM (372 mg, 455.88 mmol, 0.1 eq). The reaction mixture was then purged three times with nitrogen and reacted at 100 °C. After the reaction was complete (TLC), the mixture was extracted with water and ethyl acetate, the organic layer was evaporated to dryness, and the residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1 (v / v)) to give compound 5A (1.88 g, brown solid, yield: 49.5%).

[0236] MS(ESI): m / z 772[M+1] + .

[0237] Step 2: Synthesize compound 5B

[0238] Compound 5A (872 mg, 1.13 mmol) was dissolved in DCM (6 mL), followed by the addition of m-CPBA (390 mg, 2.26 mmol). The reaction was then carried out at 0 °C. After the reaction was complete as shown by TLC, 20 mL of saturated sodium thiosulfate aqueous solution was added to quench the reaction. The mixture was then extracted with DCM (3 × 10 mL). The organic phases were combined and washed with 15 mL of saturated NaCl solution. The organic layer was separated and dried over anhydrous sodium sulfate. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 1 (v / v)) to give compound 5B (330 mg, pale white powder, yield: 36.3%).

[0239] MS(ESI): m / z 804[M+1] + .

[0240] Step 3: Synthesize compound 5C

[0241] Compound 5B (300 mg, 0.37 mmol) and 2G (118.63 mg, 0.74 mmol) were added separately to toluene (2 mL), cooled to 0 °C in an ice bath, and then sodium tert-butoxide (35.84 mg, 0.37 mmol) was added. The reaction was then maintained at 0 °C for 5 min. After the reaction was complete as shown by TLC, the mixture was extracted with water and ethyl acetate, and the organic phase was evaporated to dryness. The residue was purified by prep-HPLC (electrolyte: EA / MeOH = 10 / 1, v / v) to give compound 5C (62 mg, yellow solid, yield: 19%).

[0242] MS(ESI): m / z 883[M+1] + .

[0243] Step 4: Synthesize compound I-5

[0244] Compound 5C (53 mg, 0.06 mmol) was added to DMF (1.0 mL), followed by cesium fluoride (42 mg, 0.28 mmol). The reaction mixture was reacted at room temperature for 1 h. The mixture was extracted with ethyl acetate, washed with brine, and dried over anhydrous sodium sulfate. The solution was then evaporated to dryness to obtain a brown oil. This brown oil was dissolved in acetonitrile (2 mL), and 8% ethyl acetate hydrochloride (1 mL) was added under ice bath conditions. The reaction mixture was reacted at room temperature for 1 h. After the reaction was complete as indicated by TLC, the reaction solution was concentrated under reduced pressure. The residue was purified by prep-HPLC to give compound I-5 (27 mg, yellow solid, 77.2%).

[0245] MS(ESI): m / z 583[M+1]+

[0246] 1 H-NMR (400MHz, DMSO) δ9.41(d,J=10.1Hz,1H),9.14(s,1H),8.27(d,J=9.4Hz,1H),7.93(d,J=6.9Hz, 1H),7.46(n,2H),7.39(d,J=2.4Hz,1H),7.12(d,J=2.5Hz,1H),5.58(d,J=53.0Hz,1H),4.65–4.55(m ,2H),4.52(d,J=13.0Hz,1H),4.38(d,J=9.1Hz,1H),4.20(s,2H),3.90(d,J=14.0Hz,1H),3.80(d,J= 13.8Hz,3H),3.65(s,1H),3.33(s,2H),2.31(d,J=15.4Hz,2H),2.17(n,2H),2.01(d,J=26.4Hz,5H).

[0247] Example 6: Preparation of compound I-6

[0248]

[0249] Synthesis route:

[0250]

[0251] Step 1: Synthesis of compound 6B

[0252] Compound 6A (600 mg, 2.03 mmol) was dissolved in DMF (6 mL), and compound 2B (473 mg, 2.23 mmol) was added. DIEA (557 mg, 4.06 mmol) was added dropwise under ice bath conditions, and the reaction was carried out at 0 °C for 30 mins. After the reaction was complete as shown by TLC, 30 mL of water was added to dilute the reaction solution. The aqueous phase was extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The organic phase was washed with saturated sodium chloride (3 × 15 mL), and then dried with anhydrous sodium sulfate. The residue was purified by silica gel column chromatography (eluent: PE:EA = 3:1 (v / v)) to give compound 6B (920 mg, pale yellow solid, 95% yield).

[0253] MS(ESI): m / z 471[M+1] + .

[0254] Step 2: Synthesize compound 6C

[0255] Compound 6B (300 mg, 0.64 mmol) was dissolved in DMF (3 mL), and 2 G (101 mg, 0.64 mmol) was added. Sodium tert-butoxide (122 mg, 1.27 mmol) was added in portions under ice bath conditions, and the reaction was carried out at 0 °C for 20 mins. After the reaction was completed as shown by TLC, 30 mL of water was added to dilute the reaction solution. The aqueous phase was extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The organic phase was washed with saturated sodium chloride (3 × 15 mL), dried over anhydrous sodium sulfate, and the residue was purified by silica gel column chromatography (eluent: EA:MEOH = 20:1 (v / v)) to give compound 6C (330 mg, pale yellow solid, yield 87%).

[0256] MS(ESI): m / z 594 [M+1] + .

[0257] Step 3: Synthesize compound 6D

[0258] Compound 6C (150 mg, 0.25 mmol) and compound 3N (120 mg, 0.25 mmol) were dissolved in dioxane (4 mL) and water (1 mL). Pd(dppf)₂Cl₂·DCM (20 mg, 0.025 mmol) and potassium carbonate (102 mg, 0.76 mmol) were added. The mixture was purged with nitrogen three times, and then the reaction solution was reacted at 90 °C for 16 hours. After the reaction was complete as shown by TLC, 30 mL of water was added to dilute the reaction solution. The aqueous phase was extracted with ethyl acetate (3 × 15 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and evaporated to dryness. The residue was purified by prep-TLC (ethyl acetate / methanol / ammonia = 20:1:0.001, v / v) to give compound 6D (40 mg, brown oil, 18% yield).

[0259] MS(ESI): m / z 882[M+1] + .

[0260] Step 4: Synthesize compound 6E

[0261] Compound 6D (40 mg, 0.05 mmol) was dissolved in DMF (2 mL), and CsF (34 mg, 0.23 mmol) was added. The reaction mixture was then reacted at room temperature for 2 hours. After the reaction was complete as shown by TLC, 30 mL of water was added to dilute the reaction mixture. The aqueous phase was extracted with ethyl acetate (3 × 15 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and evaporated to dryness to give compound 6E (30 mg, brown oil, 91% yield), which was used directly in the next step without further purification.

[0262] MS(ESI): m / z 726[M+1] + .

[0263] Step 5: Synthesize compound I-6

[0264] Compound 6E (30 mg, 0.04 mmol) was added to acetonitrile (1 mL), followed by 8% ethyl acetate hydrochloride (0.5 mL). The reaction mixture was allowed to react at room temperature for 1 h. After the reaction was complete as shown by TLC, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC to give compound I-6 (10 mg, pale yellow solid, yield 41%).

[0265] MS(ESI): m / z 582[M+1] + .

[0266] 1H NMR(400MHz,DMSO-d6)δ8.27(s,1H),7.91-7.79(m,2H),7.72-7.65(m,1H),7.56-7.42(m,3H),7 .25-7.18(m,1H),5.30-5.15(m,1H),4.29(d,J=12.2Hz,2H),4.15(s,1H),4.10(d,J=10.3Hz,1H ),4.01(d,J=10.2Hz,1H),3.64(s,1H),3.55-3.50(m,2H),3.43-3.38(m,1H),3.10-3.04(m,3H) ,2.87-2.76(m,1H),2.19-2.10(m,1H),2.09-1.86(m,3H),1.87-1.74(m,3H),1.72-1.52(m,3H).

[0267] Example 1: Determination of Phospho-ERK inhibitory activity in tumor cells

[0268] Experimental materials:

[0269] AGS cells were purchased from Nanjing Kebai; 1640 culture medium was purchased from Biological Industries; fetal bovine serum was purchased from Biosera; and Advanced Phospho-ERK1 / 2 (THR202 / TYR204) KIT was purchased from Cisbio.

[0270] The control compound BI-2852 (CAS: 2375482-51-0) was purchased from Shanghai Haoyuan Pharmaceutical Co., Ltd.

[0271] Experimental methods:

[0272] AGS cells were seeded in clear 96-well cell culture plates with 80 μL of cell suspension per well, containing 10,000 AGS cells per well. The cell culture plates were placed in a CO2 incubator and incubated overnight at 37°C.

[0273] After culturing cells overnight, remove the cell culture medium and add serum-free culture medium for serum starvation overnight.

[0274] The test compound was diluted to 2 mM with 100% DMSO as the first concentration, and then diluted 5-fold to the eighth concentration, i.e., from 2 mM to 0.026 μM, using a pipette. 2 μL of the compound was added to 78 μL of cell starvation medium, mixed well, and then 20 μL of the compound solution was added to the corresponding well of the cell plate. The cell plate was returned to the CO2 incubator for another hour. At this point, the compound concentration was 10 μM to 0.128 nM, and the DMSO concentration was 0.5%.

[0275] After incubation, discard the cell supernatant and add 50 μL of cell lysis buffer to each well, then incubate at room temperature with shaking for 30 minutes.

[0276] Dilute Eu-labeled ERK1 / 2 phosphorylated antibody and d2-labeled ERK1 / 2 phosphorylated antibody 20-fold using the detection solution; take 16 μL of cell lysate supernatant into each well of a new 384 white microplate, add 2 μL of Eu-labeled ERK1 / 2 phosphorylated antibody dilution buffer and 2 μL of d2-labeled ERK1 / 2 phosphorylated antibody dilution buffer, and incubate at room temperature for 4 hours;

[0277] After incubation, HTRF excitation was read at 320 nm and emission at 615 nm and 665 nm using a multi-label analyzer.

[0278] Data Analysis:

[0279] The original data were converted into inhibition rate using the equation (Sample-Min) / (Max-Min)*100%. The IC50 value can then be obtained by curve fitting using four parameters (obtained in GraphPad Prism using log(inhibitor) vs. response--Variable slope mode). Table 1 shows the inhibitory effect of the compounds of this invention on p-ERK. A represents activity data less than 1 μM, B represents activity data between 1 μM and 10 μM, and C represents activity data greater than 10 μM. Specific experimental results are shown in Table 1.

[0280] Table 1: IC50 data of the compound of Formula I of this invention against phosphorylated ERK (Phospho-ERK) in AGS cells.

[0281] Compound numbering IC50(nM) Reference compound BI-2852 B I-1 A I-2 A I-3 A I-4 50nM I-5 A I-6 A

[0282] Conclusion: As shown in Table 1, the compounds of this invention have a significant inhibitory effect on phosphorylation of human gastric adenocarcinoma cells with high KRAS G12D expression, and can be used as drugs for KRAS G12D inhibitors.

[0283] In the description of this specification, references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0284] Although embodiments of the present invention have been shown and described above, it is understood that these embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions, and variations to the above embodiments within the scope of the present invention without departing from its principles and spirit. It should be noted that all the above embodiments belong to the same inventive concept, and the descriptions of each embodiment have different emphases. Where the description in a particular embodiment is not exhaustive, reference can be made to the descriptions in other embodiments. These are merely preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A compound of Formula I or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof: （I） in, X is selected from CR5; Z is selected from N; Y is selected from oxygen; Y1 is selected from a single bond; L is selected from C1-C5 alkyl groups; n is selected from 0, 1, 2, 3 or 4; The R1 is selected from H, halogen, cyano, hydroxyl, -CO2R5, -CON(R5)2, C1-C3 alkyl; R2 is selected from C6-C. 12 The heterocyclic alkyl group, wherein the heterocyclic alkyl group is substituted with at least one R7 molecule; wherein the R7 molecule is independently selected from halogens, OH, CN, and NH2. The R3 is selected from aryl groups with 6-10 ring atoms, wherein the aryl group is optionally substituted by at least one R7; the R7 is independently selected from hydrogen, halogen, CN, NH2, C1-C8 alkyl, C1-C8 heteroalkyl, C1-C3 haloalkyl, C1-C3 haloalkoxy, C2-C4 alkenyl, C2-C4 alkynyl, -S-C1-C8 alkyl, -O-C1-C8 alkyl; The R4 is selected from H, halogen, C1-C3 alkyl, C3-C8 cycloalkyl, C1-C3 alkoxy or C1-C3 haloalkoxy; The R5 is independently selected from hydrogen, OH, CN, NH2, halogen, C1-C3 alkyl, C3-C8 cycloalkyl, C1-C3 alkoxy or C1-C3 haloalkoxy.

2. The following compounds, or their pharmaceutically acceptable salts, stereoisomers, and tautomers, ， 。 3. A pharmaceutical composition comprising an effective dose of the compound as described in any one of claims 1 to 2 or a pharmaceutically acceptable salt, stereoisomer, tautomer thereof, and at least one pharmaceutically acceptable excipient.

4. Use of a compound as described in any one of claims 1 to 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or pharmaceutical composition of claim 3, in the preparation of a medicament for treating diseases caused by KRAS G12D mutations.

5. Use of a compound as described in any one of claims 1 to 2, or a pharmaceutically acceptable salt, stereoisomer, tautomer, or pharmaceutical composition of claim 3, in the preparation of a KRAS G12D inhibitor medicament.

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