Hydroxyl-substituted dihydroisoquinolinone compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry, and particularly relates to a hydroxyl-substituted dihydroisoquinolinone compound. The compound has a good KRAS G12V mutant gene inhibition effect, and can be used for treating tumor diseases related to KRAS G12V gene mutation.

Description

[0001] This application claims priority to Chinese patent application No. 202310809298X filed on July 3, 2023. Technical Field

[0002] The present invention belongs to the technical field of pharmaceutical chemistry, and specifically relates to a class of hydroxy-substituted dihydroisoquinolinone compounds. The compounds have good KRAS G12V mutation gene inhibition effects and can be used for tumor diseases mediated by KRAS G12V gene mutations. Background Art

[0003] The RAS gene is one of the most commonly mutated genes in cancer cells, with RAS gene mutations detected in 30% of human tumors. The RAS subfamily includes HRAS, NRAS, and KRAS, with KRAS mutations being the most common in cancer cells, accounting for about 85%. KRAS mutations and / or KRAS wild-type amplification are common in colorectal cancer (about 45% in the United States and about 49% in China), pancreatic cancer (about 90% in the United States and about 87% in China), and non-small cell lung cancer (subtype adenocarcinoma: about 35% in the United States and about 13% in China). KRAS mutations have also been found in some cancers, including gastric cancer, uterine cancer, cervical cancer, bladder cancer, bile duct cancer, diffuse large B-cell lymphoma, multiple myeloma, and squamous cell carcinoma of the skin. The distribution of KRAS alleles varies by tumor type, with G12D (41%), G12C (14%) and G12V (28%) being the most common. G12C is mutated in 13.6% of all lung adenocarcinomas, while G12D and G12V are the two most common alleles in colorectal cancer and pancreatic cancer.

[0004] KRAS protein is a class of guanine nucleotide binding proteins with GTP hydrolase activity, acting as a molecular switch in a series of intracellular signaling pathways. The balance of nucleotide hydrolysis and exchange determines the level of KRAS in the activated state in the cell. When bound to GDP, KRAS is in the "OFF" state, while under the action of guanylate exchange factors (such as SOS1), GTP is exchanged with GDP, which puts KRAS in the "ON" state. In this state, KRAS can activate downstream signaling pathways, including mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) pathways, thereby promoting cell proliferation and survival. In cancer cells, mutations in KRAS disrupt the guanine-promoted exchange process, usually by becoming GAP-independent and "locking" RAS in an active GTP-bound state, thereby activating downstream signaling pathways for tumor cell growth.

[0005] On the one hand, the affinity of KRAS for GTP is at the picomolar level, while the intracellular concentration of GTP is as high as 0.5 μM. Therefore, it is difficult for small molecules to effectively compete with GTP. On the other hand, because the surface of KRAS proteins is relatively smooth, they have a nearly spherical structure, no deep hydrophobic pockets, and no obvious binding sites. Therefore, in the past few decades, targeting KRAS has been considered very challenging. In recent years, candidate compounds targeting KRAS G12C have been developed one after another. As the world's first drug targeting KRAS G12C, Amgen's Sotorasib (AMG510) was approved for marketing by the FDA in May 2021. However, there are currently no candidate molecules on the market for other mutations, so developing KRAS inhibitors for other subtypes of mutations as soon as possible and pushing them into the clinical stage has also become a key development area. Summary of the invention

[0006] The first aspect of the present invention provides a compound represented by formula I or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof;

[0007]

[0008] Wherein, X1, X2, and X3 are selected from N and C-Ra; Ra is selected from H, halogen atoms, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy;

[0009] L1 is selected from O, S, NH;

[0010] Ring B is a 5-6 membered aryl group or a 5-6 membered heteroaryl group, and Ring B may be optionally substituted by one or more groups selected from the following: F, Cl, Br, CN, OH, NH2, C 1-3 Alkyl, C 1-3 Haloalkyl;

[0011] R1 is a 6-12 membered nitrogen-containing hetero-bridged ring substituted C 1-3 The nitrogen-containing hetero bridged ring is a bicyclic bridged ring and may be optionally substituted by one or more groups selected from the following: F, Cl, CN, NH2, OH, C 1-3 Alkyl, C 1-3 Haloalkyl;

[0012] R2 is selected from H, CN, halogen atoms, C 1-3 Alkyl, C 1-3 Haloalkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, hydroxy-C 1-3 alkyl;

[0013] R3 is selected from H, CN, halogen atoms, C 1-3 Alkyl, C1-3 Haloalkyl, C 1-3 Alkoxy, Hydroxyl-C 1-3 alkyl;

[0014] R x R5 is selected from C 1-3 alkyl, R5 substituted 3-7 membered cycloalkyl, R5 substituted 3-7 membered heterocycloalkyl, R5 substituted C6-C 10 aryl, 5-6 membered heteroaryl substituted by R5; R5 is selected from H, CN, OH, halogen atom, C 1-3 Alkyl, C 1-3 Haloalkyl, C 3-6 Cycloalkyl, C 3-6 Heterocycloalkyl;

[0015] Or R3, Rx and the carbon atom to which they are connected are cyclized together to form a 5-6-membered aryl or 5-6-membered heteroaryl; the 5-6-membered aryl or the 5-6-membered heteroaryl may be optionally replaced by R6 and R 6a Substituted C 1-3 alkyl; R6 is selected from H, 5-6 membered aryl, 5-6 membered heteroaryl; and R6 may be optionally substituted by one or more of the following groups: H, NH2, CN, halogen atom, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, Hydroxyl-C 1-3 Alkyl; said R 6a Selected from C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, Hydroxyl-C 1-3 alkyl.

[0016] In one technical solution of formula I of the present invention, ring B is substituted by OH; or ring B is substituted by Cl;

[0017] In any of the above technical solutions of formula I of the present invention, ring B is substituted by OH and Cl at the same time;

[0018] In any of the above technical solutions of formula I of the present invention, at least one of X1 and X2 is N; or both X1 and X2 are N;

[0019] In any of the above technical solutions of Formula I of the present invention, X3 is N or C-Ra; preferably X3 is N; Ra has the same meaning as in the above Formula I, preferably H, C 1-3 alkyl;

[0020] In any of the above technical solutions of formula I of the present invention, X1, X2, and X3 are all N;

[0021] In any of the above technical solutions of formula I of the present invention, L1 is O, or L1 is S, or L1 is NH;

[0022] In any of the above technical solutions of formula I of the present invention, ring B is a benzene ring, and may be optionally substituted by Cl or / and OH;

[0023] In any of the above technical solutions of formula I of the present invention, R1 is the following group substituted by one or more halogen atoms; Preferably, R1 is selected from the following groups:

[0024] In any of the above technical solutions of formula I of the present invention, R2 is selected from methyl, ethyl, isopropyl, and cyclopropyl;

[0025] In any of the above technical solutions of formula I of the present invention, R3 is H;

[0026] In any of the above technical solutions of formula I of the present invention, It has the following structure:

[0027] In any of the above technical solutions of formula I of the present invention, the 5-6 membered heteroaryl group formed by the cyclization of R3, Rx and the carbon atom to which they are connected is selected from imidazole and pyrrole;

[0028] In any of the above technical solutions of formula I of the present invention, The specific structure is as follows: Among them, R6, R 6a has the same meaning as in Formula I; X4 is N or C-Ra, Ra has the same meaning as in Formula I; preferably, X4 is N or CH; R 6a is selected from H, methyl, trifluoromethyl; R 6a is selected from pyridyl and pyrazolyl; in one technical solution, R6 is selected from 1-methylpyrazol-2-yl and 2-aminopyridin-3-yl;

[0029] In any of the above technical solutions of formula I of the present invention, the 5-6 membered aryl group in R6 is a phenyl group;

[0030] In any of the above technical solutions of formula I of the present invention, the 5-6 membered heteroaryl group in R6 is selected from pyridine and pyrazolyl;

[0031] In any of the above technical solutions of formula I of the present invention, R6 is selected from 2-amino-pyridin-3-yl and 1-methylpyrazol-3-yl.

[0032] In a technical solution of Formula I of the present invention, the compound represented by Formula I is preferably a compound of the following general structure:

[0033]

[0034] In any of the above technical solutions of Formula I of the present invention, the compound represented by Formula I is selected from the following compounds or pharmaceutically acceptable salts thereof;

[0035]

[0036]

[0037] The second aspect of the present invention provides a pharmaceutical composition comprising a compound represented by Formula I or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, and any pharmaceutically acceptable carrier.

[0038] The third aspect of the present invention provides use of the compound of formula I or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof in the preparation of a drug for treating a disease associated with the KRAS G12V gene mutation.

[0039] The fourth aspect of the present invention provides use of the pharmaceutical composition described in the second aspect of the present invention in the preparation of a drug for treating diseases associated with KRASG12V gene mutation.

[0040] In a fifth aspect, the present invention provides a method for treating a disease associated with the KRAS G12V gene mutation, the method comprising administering to a patient in need thereof an effective amount of the compound of formula I or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or the pharmaceutical composition of the second aspect of the present invention.

[0041] In the present invention, the disease associated with KRAS G12V gene mutation may be cancer associated with KRAS G12V gene mutation.

[0042] Definition and Explanation

[0043] In the present invention, for some compounds of the present invention, such as compound 12, when a chiral center has a single stereo configuration but the specific absolute configuration cannot be determined, its adjacent atoms are marked with wavy solid lines. Indicates the chemical bond. For example, the chiral atom to which the ethyl group in compound 12 is connected is a single stereo configuration obtained after chiral separation, but the stereo configuration has not yet been determined, that is, " express;

[0044] In the present invention, some of the compounds of the present invention, such as compound 15, may be separated as formate because the mobile phase of preparative HPLC contains formic acid.

[0045] In the present invention, especially in the general formula of the compound of formula I and its sub-compounds, the H refers to the hydrogen element, that is, an atom with a nucleus having 1 proton, which includes three isotopes of protium (P), deuterium (D), and tritium (T);

[0046] In the present invention, the "halogen" or "halogen atom" refers to fluorine, chlorine, bromine, and iodine; the "halo" refers to a group formed after one or more hydrogen atoms in a substituent are replaced by a halogen atom.

[0047] The term "alkyl" refers to a straight or branched hydrocarbon group in which carbon atoms and carbon atoms and hydrogen atoms are connected by single bonds. 1-3 or C 1-6 Alkyl; C 1-3 "Alkyl" means a straight or branched chain alkyl group having 1 to 3 carbon atoms; 1-3 Specific examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl; "C 1-6 "Alkyl" means a straight or branched chain alkyl group having 1 to 6 carbon atoms. 1-6 Examples of alkyl groups include but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl.

[0048] The term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by halogen atoms. 1-3 Haloalkyl, C 1-6 Halogenated alkyl. 1-3 Examples of haloalkyl include, but are not limited to, monofluoromethyl, monochloromethane, difluoromethyl, dichloromethane, trifluoromethyl, trichloromethyl, tribromomethyl.

[0049] The term "alkoxy" refers to an alkyl-O-alkyl group, wherein the alkyl group is as described above and includes C 1-3 Alkoxy or C 1-6 Alkoxy; C 1-3 Specific examples of alkoxy include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy.

[0050] The term "haloalkoxy" refers to an alkoxy group in which one or more hydrogen atoms are replaced by a halogen atom. 1-3 Examples of haloalkoxy include, but are not limited to, trifluoromethoxy, trichloromethoxy, 2,2,2-trifluoroethoxy, 2,2,2-trichloroethoxy.

[0051] The term "cycloalkyl" refers to a cyclic saturated monocyclic or polycyclic ring formed by connecting carbon atoms by single bonds. The ring C atoms are optionally oxo-substituted, i.e., -CH2- is oxo-substituted to form -C(O)-. Specific examples of 3-6-membered cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Specific examples of 3-7-membered cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0052] The term "heterocyclyl" or "heterocycloalkyl" refers to monocyclic and polycyclic groups derived from the replacement of one or more ring carbon atoms in a cycloalkyl group by one or more heteroatoms or heteroatomic groups; the heteroatoms or heteroatomic groups are generally selected from N, N(O), O, S, S(O), S(O)2; the ring C atoms are optionally oxo-substituted, i.e. -CH2- is oxo-substituted to form -C(O)-. Cycloalkyl is as defined above. Specific examples of 3-6 membered heterocycloalkyl groups include oxirane, azetidinyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl; specific examples of 3-7 membered heterocycloalkyl groups include oxirane, azetidinyl, tetrahydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, azepanyl;

[0053] The term "bridged ring" refers to a ring structure formed by two adjacent rings sharing two ring carbon atoms; the term "heterobridged ring" refers to a bridged ring formed by replacing one or more bridged ring atoms of the bridged ring with atoms selected from N, O, and S. Specific examples of 6-12 membered nitrogen-containing heterobridged rings include

[0054] The term "aryl" refers to an unsaturated, usually aromatic hydrocarbon group, which may be a single ring or multiple rings fused together. Examples of aryl include, but are not limited to, phenyl and naphthyl. A specific example of a 5-6 membered aryl group is phenyl.

[0055] The term "heteroaryl" refers to a monocyclic or polycyclic aromatic group in which at least one ring atom is a heteroatom, and the heteroatom is selected from N, O, S. When the heteroaryl contains an N atom, it also includes its nitrogen oxide. The polycyclic heteroaryl is a fused heteroaryl, which includes but is not limited to benzo 5-6 membered heteroaryl, 5-6 membered heteroaryl and 5-6 membered heteroaryl. The heteroaryl includes 5-6 membered heteroaryl, 7-10 membered bicyclic fused heteroaryl, and 8-14 membered fused heteroaryl. Examples of heteroaryl include but are not limited to pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isoxazolyl, pyridyl, pyrimidinyl, indazolyl, indolyl, isoquinolyl, quinoxalinyl, benzoxazolyl, benzofuranyl, benzothienyl, benzothiazolyl, benzimidazolyl, quinolyl, and quinazolinyl. Examples of 5-6 membered heteroaryl groups include pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, pyrazinyl, pyridazinyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isoxazolyl, pyridinyl, and pyrimidinyl. Examples of 7-10 membered bicyclic fused heteroaryl groups include indolyl, isoquinolyl, quinoxalinyl, benzoxazolyl, benzofuranyl, benzothienyl, benzothiazolyl, benzimidazolyl, quinolyl, and quinazolinyl.

[0056] The term "optionally" means that the substituent may be substituted by other substituents, or may not be substituted by other substituents.

[0057] The term "composition" is intended to include products comprising the specified amounts of each specified ingredient, as well as any product resulting directly or indirectly from the combination of the specified amounts of each specified ingredient. Those skilled in the art can modify the actual dosage levels of each active ingredient in the pharmaceutical composition of the present invention so that the amount of active compound obtained can effectively obtain the desired therapeutic response for a specific patient, composition and administration method.

[0058] The term "pharmaceutically acceptable carrier" refers to a medium generally accepted in the art for delivering biologically active agents to animals, especially mammals.

[0059] The term "excipient" generally refers to a carrier, diluent and / or vehicle required to formulate an effective pharmaceutical composition.

[0060] The term "effective amount" refers to a compound of the present invention or a pharmaceutically acceptable salt thereof, which is a sufficient amount of the compound to treat the disorder at a reasonable effect / risk ratio applicable to any medical treatment and / or prevention.

[0061] The term "pharmaceutically acceptable salt" refers to salts that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic response, etc., and are commensurate with a reasonable benefit / risk ratio.

[0062] In the present invention, the substituent Indicates the position of attachment of the substituent to the parent structure or other fragments. The presence of a dash "-" in a substituent structure indicates the point of attachment for the substituent, for example -CH3 is attached through a C atom. and Indicates the absolute configuration of a stereocenter, i.e., R or S configuration.

[0063] The "isomers" described in the present invention include geometric isomers and stereoisomers, such as atropisomers, cis-trans isomers, enantiomers, diastereomers, tautomers, and racemic mixtures and other mixtures thereof, all of which are within the scope of the present invention. The term "enantiomer" refers to stereoisomers that are mirror images of each other. The term "tautomer" refers to a type of functional group isomer that has different hydrogen attachment points through one or more double bond displacements, for example, a ketone and its enol form are keto-enol tautomers. The term "diastereomer" refers to a stereoisomer in which a molecule has two or more chiral centers and the molecules are non-mirror images. The term "cis-trans isomer" refers to different spatial configurations in which double bonds or single bonds of ring carbon atoms in a molecule cannot rotate freely. The term "atropisomer" refers to a stereoisomer that can be separated because single bond rotation is hindered or rotates very slowly.

[0064] Stereoisomers of the compounds of the present invention can be prepared by chiral synthesis or chiral reagents or other conventional techniques. For example, one enantiomer of a compound of the present invention can be prepared by asymmetric catalysis technology or chiral auxiliary derivatization technology. Alternatively, a compound of a single stereo configuration can be obtained from a mixture by chiral resolution technology. Alternatively, it can be directly prepared using chiral starting materials. The separation of optically pure compounds in the present invention is usually completed using preparative chromatography, using a chiral chromatographic column to achieve the purpose of separating chiral compounds.

[0065] The chemical abbreviations used in the present invention and the chemical names they refer to are as follows:

[0066] DETAILED DESCRIPTION

[0067] The present invention is further described in detail below through specific preparation examples and biological experiments, but it should be understood that these examples and biological experiments are only used for specific illustration purposes and should not be understood as limiting the present invention in any form. It is clear to those skilled in the art that, hereinafter, if not specifically stated, the materials used are well known in the art and can be purchased from the market or obtained by those skilled in the art according to published literature or conventional methods. Unless otherwise specified, all reactions of the present invention are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, and the solvent is a dry solvent, wherein: (i) the temperature is expressed in degrees Celsius (°C), and the operation is carried out at room temperature, which generally refers to 15-35°C, preferably 20-30°C, and more preferably 20-25°C; (ii) the removal of the solvent is carried out by reduced pressure evaporation on a rotary evaporator, and the bath temperature is generally not higher than 60°C; (iii) the reaction process is tracked by thin layer chromatography (TLC); (iv) the final product has a satisfactory hydrogen nuclear magnetic resonance spectrum ( 1 H-NMR) and / or mass spectrometry (MS) data.

[0068] Test equipment:

[0069] The structure of the compound of the present invention is determined by nuclear magnetic resonance (NMR) or / and liquid chromatography-mass spectrometry (LC-MS). The NMR chemical shift (δ) is given in parts per million (ppm). The NMR is measured using a Bruker Neo 400M or Bruker Ascend 400 nuclear magnetic instrument, and the measurement solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD) and / or deuterated chloroform (CDCl3), and the internal standard is tetramethylsilane (TMS).

[0070] Liquid chromatography-mass spectrometry (LC-MS) was performed using a Shimadzu 2030 Plus-LCMS 2020 mass spectrometer, an Agilent 1260-6125B single quadrupole mass spectrometer, or a Shimadzu LCMS-2020 mass spectrometer. HPLC was performed using a Shimadzu LCMS-2020 or Agilent 1260 high performance liquid chromatograph.

[0071] Preparative HPLC was performed using a Shimadzu FRC-40 equipped with LC-20AP and PDA-20A (chromatographic column: Synergi Max-RP, 150×30 mm, 4 m) or a GILSON Trilution LC (chromatographic column: SunFirePrep C18, 10 μm, 19×250 mm XBridge Prep C18, 10 μm, 19×250 mm)).

[0072] The model of the microwave synthesizer is Biotage Initiator+.

[0073] Example 1 and Example 2: Preparation of Compound 1 and Compound 2

[0074]

[0075] Step 1: Synthesis of compound 1-2

[0076]

[0077] Compound 1-1 (1.5 g, 8.26 mmol) was dissolved in tetrahydrofuran (20 mL), and then a tetrahydrofuran solution of lithium bis(trimethylsilyl)amide (8.67 mL, 1 mol / L) was slowly added to the reaction solution under a nitrogen atmosphere at -65 °C, and then reacted at -65 °C for 1 hour. Ethyl iodide (1.35 g, 8.67 mmol) was added, and then reacted at -65 °C for 1 hour. 1M dilute hydrochloric acid (50 mL) was added, and extracted with ethyl acetate (20 mL × 3), and the organic phases were combined and washed with saturated brine (50 mL), and the organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 1-2. MS m / z (ESI): 183.0 [M-CN] + .

[0078] Step 2: Synthesis of Compound 1-3

[0079]

[0080] Compound 1-2 (1.5 g, 7.15 mmol) was dissolved in methanol (10 mL), and aqueous ammonia (0.14 mL) and Raney nickel (4.69 g) were added to replace the hydrogen, and then reacted at 25° C. for 18 hours under a hydrogen atmosphere, filtered, and the filtrate was concentrated under reduced pressure and the crude product was purified by rapid silica gel column chromatography (eluent: dichloromethane / methanol) to obtain compound 1-3. MS m / z (ESI): 214.2 [M+H] + .

[0081] 1 H NMR(400MHz,DMSO-d6)δ:7.21(d,J=8.8Hz,1H),6.99(d,J=2.8Hz,1H),6.91(dd,J=8.8,2.8Hz,1H) ,3.81(s,3H),2.91-3.06(m,1H),2.70(d,J=6.8Hz,2H),1.12-1.96(m,4H),0.72(t,J=7.2Hz,3H).

[0082] Step 3: Synthesis of Compound 1-4

[0083]

[0084] Dissolve triphosgene (666 mg, 2.25 mmol) in dichloromethane (10 mL), cool the reaction solution to 0°C under nitrogen protection, add compound 1-3 (800 mg, 3.74 mmol) and diisopropylethylamine (483 mg, 3.74 mmol), and react for 1 hour. Concentrate the reaction solution under reduced pressure, add water (10 mL) to dilute, extract with ethyl acetate (20 mL×3), dry the organic phase with anhydrous sodium sulfate, filter and concentrate to obtain compound 1-4. MS m / z (ESI): 240.0 [M+H] + .

[0085] Step 4: Synthesis of Compound 1-5

[0086]

[0087] Compound 1-4 (830 mg, 3.46 mmol) was dissolved in trifluoromethanesulfonic acid (10 mL), heated to 100 ° C and stirred for 1 hour. The temperature was lowered to 0 ° C, saturated sodium bicarbonate solution was added dropwise to the reaction solution, the pH of the reaction solution was adjusted to 8-9, water (30 mL) was added to dilute, and extracted with ethyl acetate (30 mL × 3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by rapid silica gel column chromatography (eluent: dichloromethane / methanol) to obtain compound 1-5. MS m / z (ESI): 226.0 [M+H] + .

[0088] Step 5: Synthesis of Compound 1-6

[0089]

[0090] Compound 1-5 (300 mg, 1.33 mmol) was dissolved in acetonitrile (5 mL), potassium carbonate (551.2 mg, 3.99 mmol) and benzyl bromide (295.6 mg, 1.73 mmol) were added, and the temperature was raised to 50°C for reaction for 18 hours. The mixture was cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (15 mL × 3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 1-6. MS m / z (ESI): 316.2 [M+H] + .

[0091] Step 6: Synthesis of Compound 1-8

[0092]

[0093] Under a nitrogen atmosphere at 0°C, diisopropyl azodicarboxylate (0.42 mL, 2.12 mmol) was added to a solution of compound 1-7 (200 mg, 1.06 mmol), 3-(benzyloxy)cyclobutanol (188.60 mg, 1.06 mmol) and triphenylphosphine (555.11 mg, 2.12 mmol) in anhydrous tetrahydrofuran (5 mL). After the addition, the mixture was heated to 70°C and stirred for 18 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 1-8. MS m / z (ESI): 349.0 [M+H] + .

[0094] Step 7: Synthesis of Compound 1-9

[0095]

[0096] Compound 1-6 (250 mg, 0.79 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (31.6 mg, 0.79 mmol, purity 60%) was added at 0°C, then stirred for 1 hour, compound 1-8 (414 mg, 1.19 mmol) was added, and the mixture was reacted at 0°C for 1 hour. The reaction solution was added dropwise to a saturated ammonium chloride solution (5 mL) to quench, diluted with water (10 mL), extracted with ethyl acetate (20 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 1-9. MS m / z (ESI): 628.2 [M+H] +.

[0097] Step 8: Synthesis of Compound 1-10 and Compound 1-11

[0098]

[0099] Compound 1-9 (150 mg, 0.24 mmol) was dissolved in dioxane (2 mL), ((2R, 7aS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (57 mg, 0.36 mmol), palladium acetate (5.4 mg, 0.02 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (27.6 mg, 0.05 mmol), cesium carbonate (233.3 mg, 0.72 mmol) were added, and after replacing nitrogen, the mixture was reacted at 130°C in a microwave reactor for 10 hours. Water (10 mL) was added for dilution, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified using a fast silica gel column (eluent: eluent: dichloromethane / methanol) to obtain compound 1-10, MS m / z (ESI): 751.4 [M+H] + ; Compound 1-11, MS m / z (ESI): 717.4 [M+H] + .

[0100] Step 9: Synthesis of compound 1

[0101]

[0102] Compound 1-10 (40 mg, 0.05 mmol) was dissolved in trifluoroacetic acid (3 mL) and heated to 90 °C for 7 hours. The reaction solution was decompressed to remove the solvent and purified by preparative HPLC (chromatographic column: Xtimate C18, 150×40 mm, 5 mm; mobile phase: 0.075% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile 15%-45%, 15 min) to obtain compound 1 (trifluoroacetate). MS m / z (ESI): 571.3 [M+H] + .

[0103] 1H NMR (400MHz, DMSO-d6) δ: 10.80 (s, 1H), 10.40-10.07 (m, 1H), 8.55 (s, 1H), 7.41 (d, J = 2.4Hz, 1H), 7.15 (d, J = 2. 4Hz,1H),5.79-4.94(m,3H),4.67-4.51(m,3H),4.21-4.15(m,2H),3.95-3.82(m,2H),3.82-3.74(m,2H),3.32 (d,J=4.0Hz,1H),3.14(d,J=2.0Hz,1H),2.90-2.78(m,2H),2.69-2.58(m,1H),2.45(d,J=3.6Hz,1H),2.42-2. 40(m,1H),2.38-2.31(m,1H),2.25-2.11(m,2H),2.11-2.01(m,1H),1.74-1.53(m,2H),0.87(t,J=7.2Hz,3H).

[0104] Step 10: Synthesis of Compound 2

[0105]

[0106] Compound 1-11 (40 mg, 0.05 mmol) was dissolved in trifluoroacetic acid (3 mL) and heated to 90 °C for 7 hours. The reaction solution was decompressed to remove the solvent and purified by preparative HPLC (chromatographic column: Xtimate C18, 150×40 mm, 5 mm; mobile phase: 0.075% trifluoroacetic acid aqueous solution-acetonitrile; gradient: acetonitrile 15%-45%, 15 min) to obtain compound 2 (trifluoroacetate). MS m / z (ESI): 537.3 [M+H] + .

[0107] 1H NMR(400MHz,DMSO-d6)δ:10.81-10.60(m,1H),9.73(s,1H),8.54(s,1H),7.40(d,J=2.4Hz,1H),7.29-7 .21(m,1H),7.02(dd,J=2.8,8.4Hz,1H),5.74-5.08(m,3H),4.68-4.49(m,3H),4.23-4.07(m,2H),3.94- 3.81(m,2H),3.80-3.71(m,2H),3.36-3.30(m,1H),2.92-2.81(m,3H),2.71-2.59(m,1H),2.46-2.42(m, 2H),2.40-2.32(m,1H),2.26-2.14(m,2H),2.11-1.99(m,1H),1.73-1.58(m,2H),0.85(t,J=7.2Hz,3H).

[0108] Example 3: Preparation of Compound 3

[0109]

[0110] Step 1: Synthesis of compound 3-2

[0111]

[0112] Compound 3-1 (5.0 g, 34.0 mmol) was dissolved in tetrahydrofuran (100 mL), and lithium bis(trimethylsilyl)amide (18.7 mL, 37.4 mmol, 2 mol / L) was added at -78°C under nitrogen atmosphere, and then stirred for 1 hour, and iodoisopropyl (6.36 g, 37.4 mmol) was added, and the temperature was raised to -40°C for reaction for 1 hour. The temperature was lowered to -78°C, and the mixture was added dropwise to a saturated ammonium chloride solution (100 mL), and diluted with water (50 mL), and extracted with ethyl acetate (100 mL×3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 3-2.

[0113] 1 HNMR(400MHz, CDCl3)δ:7.24–7.17(m,2H),6.92–6.86(m,2H),3.81(s,3H),3. 60(d,J=6.4Hz,1H), 2.09(dt,J=13.2,6.8Hz,1H), 1.03(dd,J=6.8,2.0Hz,6H).

[0114] Step 2: Synthesis of compound 3-3

[0115]

[0116] Compound 3-2 (4.0 g, 21.1 mmol) was dissolved in methanol (50 mL), and aqueous ammonia (1 mL) and Raney nickel-water suspension (4.0 mL) were added to replace the hydrogen gas. The mixture was stirred for 18 hours under a hydrogen atmosphere, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified using a fast silica gel column (eluent: eluent: dichloromethane / methanol) to obtain compound 3-3. MS m / z (ESI): 194.2 [M+H] + .

[0117] 1 HNMR(400MHz, DMSO-d6)δ:7.08–7.04(m,2H),6.86(d,J=8.4Hz,2H),3.73(s,3H),2.87(dd,J=12.4,5.2Hz,1H),2.73(dd ,J=12.4,9.2Hz,1H),2.24(ddd,J=8.8,7.6,5.2Hz,1H),1.93–1.73(m,1H),0.89(d,J=6.8Hz,3H),0.64(d,J=6.8Hz,3H).

[0118] Step 3: Synthesis of compound 3-4

[0119]

[0120] Dissolve triphosgene (2.1 g, 7.2 mmol) in dichloromethane (30 mL), add compound 3-3 (2.0 g, 10.4 mmol) and diisopropylethylamine (4.0 g, 31.2 mmol) at 0°C under nitrogen atmosphere, and stir for 1 hour. Concentrate the reaction solution under reduced pressure, add water (50 mL) to dilute, extract with ethyl acetate (100 mL×3), dry the organic phase with anhydrous sodium sulfate, filter and concentrate to obtain compound 3-4. MS m / z (ESI): 220.2 [M+H] + .

[0121] Step 4: Synthesis of Compound 3-5

[0122]

[0123] Compound 3-4 (2.0 g, 9.1 mmol) was dissolved in trifluoromethanesulfonic acid (10 mL), heated to 100 ° C and stirred for 1 hour. The mixture was cooled to 0 ° C and added dropwise to saturated sodium bicarbonate (20 mL), diluted with water (30 mL), extracted with ethyl acetate (100 mL × 3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by rapid silica gel column chromatography (eluent: dichloromethane / methanol) to obtain compound 3-5. MS m / z (ESI): 206.2 [M+H] + .

[0124] 1 HNMR(400MHz, Methanol-d4)δ:7.24(d,J=2.8Hz,1H),7.01(d,J=8.4Hz,1H),6.83(dd,J=8.4,2.8Hz,1H),3.45(t, J=3.2Hz,2H),2.34(ddd,J=7.2,4.0,2.8Hz,1H),1.84–1.73(m,1H),0.91(d,J=6.8Hz,3H),0.77(d,J=6.8Hz,3H).

[0125] Step 5: Synthesis of Compound 3-6

[0126]

[0127] Compound 3-5 (200 mg, 0.97 mmol) was dissolved in acetonitrile (5 mL), potassium carbonate (403 mg, 2.92 mmol) and benzyl bromide (200 mg, 1.17 mmol) were added, and the temperature was raised to 50°C for reaction for 16 hours. Cooled to room temperature, water (25 mL) was added for dilution, and extracted with ethyl acetate (25 mL×2). The organic phase was washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 3-6. MS m / z (ESI): 296.2 [M+H] + .

[0128] Step 6: Synthesis of Compound 3-7

[0129]

[0130] Compound 3-6 (160 mg, 0.54 mmol) was added to a suspension of sodium hydride (23.8 mg, 0.60 mmol, purity 60%) in N,N-dimethylformamide (1 mL) at 0°C. The reaction solution was stirred at 0°C for 5 minutes. A solution of compound 1-8 (198.6 mg, 0.57 mmol) in N,N-dimethylformamide (1 mL) was added, and the reaction solution was stirred at 0°C for 25 minutes. The reaction solution was poured into 100 mL of water for quenching and extracted with ethyl acetate (20 mL×2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. Compound 3-7 was obtained by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether). MS m / z(ESI):608.4[M+H] + .

[0131] Step 7: Synthesis of Compound 3-8

[0132]

[0133] Compound 3-7 (100 mg, 0.16 mmol) and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (78.5 mg, 0.49 mmol) were dissolved in N,N-dimethylformamide (3 mL), and cesium carbonate (160.7 mg, 0.49 mmol), palladium acetate (5.5 mg, 0.02 mmol) and 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (10.2 mg, 0.02 mmol) were added under nitrogen protection. The reaction solution was heated to 110 ° C and stirred for 2 hours under nitrogen atmosphere protection. The reaction solution was poured into 50 mL of water to quench, and extracted with ethyl acetate (10 mL×2). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness under reduced pressure to obtain a crude product. The crude product was purified by flash silica gel column chromatography (eluent: methanol / dichloromethane) to obtain compound 3-8. MS m / z (ESI): 731.4 [M+H] + .

[0134] Step 8: Synthesis of compound 3

[0135]

[0136] Compound 3-8 (120 mg, 0.16 mmol) was dissolved in trifluoroacetic acid (3 mL), and the reaction solution was stirred at 90 ° C for 6 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain a crude product. Compound 3 was separated by preparative high performance liquid chromatography (chromatographic column: Waters-Xbridge-C18, 19×250 mm, 10 μm, mobile phase: 10 mM ammonium bicarbonate aqueous solution-acetonitrile, gradient: acetonitrile 20%-50%, flow rate: 30 mL / min). MS m / z (ESI): 551.7 [M+H] + .

[0137] 1 H NMR (400MHz, DMSO-d6) δ: 9.77 (s, 1H), 8.45 (s, 1H), 7.38 (d, J = 2.8Hz, 1H), 7.20 (d, J = 8. 4Hz,1H),7.04-6.95(m,1H),5.50-5.30(m,2H),5.22-5.11(m,1H),4.58-4.48(m,1H),4 .35-4.08(m,5H),3.04-2.96(m,1H),2.88-2.77(m,2H),2.68-2.61(m,1H),2.48-2.43( m,3H),2.42-2.18(m,3H),2.17-2.08(m,1H),2.02-1.82(m,4H),0.83(d,J=6.8Hz,6H).

[0138] Example 4: Preparation of Compound 4

[0139]

[0140] Step 1: Synthesis of compound 4-1

[0141]

[0142] Compound 1-7 (2.0 g, 10.6 mmol) was dissolved in N, N-dimethylformamide (20 mL), sodium hydride (520 mg, 13.0 mmol, purity 60%) was added at 0°C under nitrogen atmosphere, then stirred for 1 hour, iodomethane (1.56 g, 11.0 mmol) was added, the temperature was raised to 25°C, and the reaction was continued for 1 hour. The mixture was cooled to 0°C, added dropwise to a saturated ammonium chloride solution (100 mL), diluted with water (50 mL), extracted with ethyl acetate (80 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 4-1. MS m / z(ESI):203.0[M+H] + .

[0143] Step 2: Synthesis of compound 4-2

[0144]

[0145] Sodium hydride (46.1 mg, 1.15 mmol, purity 60%) was added to a solution of compound 3-6 (170 mg, 0.58 mmol) in N, N-dimethylformamide (3 mL) at 0°C, stirred at 0°C for 30 minutes, and compound 4-1 (140.2 mg, 0.69 mmol) was added, and the reaction was continued at 0°C for 2.5 hours. The reaction solution was quenched with saturated ammonium chloride solution (25 mL), extracted with ethyl acetate (50 mL × 3), and the combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain 4-2. MS m / z (ESI): 462.2 [M + H] + .

[0146] Step 3: Synthesis of compound 4-3:

[0147]

[0148] 4-2 (80 mg, 0.17 mmol), ((2R, 7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (82.7 mg, 0.52 mmol), 1,1'-binaphthyl-2,2'-bisdiphenylphosphine (10.8 mg, 0.02 mmol), cesium carbonate (169.3 mg, 0.52 mmol), N,N-dimethylformamide (2 mL) were added to an 8 ml reaction bottle, and palladium acetate (5 mg, 0.02 mmol) was added after purging nitrogen. The mixture was reacted at 110 ° C for 2 hours under nitrogen atmosphere. The reaction mixture was quenched with saturated ammonium chloride solution (25 mL). It was extracted with ethyl acetate (50 mL × 3), and the combined organic layer was washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by rapid silica gel column chromatography (methanol / dichloromethane) to obtain 4-3. MSm / z(ESI):585.3[M+H] + .

[0149] Step 4: Synthesis of compound 4

[0150]

[0151] Palladium hydroxide (20 mg, 0.01 mmol, purity 10%) and palladium carbon (30 mg, 0.01 mmol, purity 10%) were added to a methanol (5 mL) solution of compound 4-3 (75 mg, 0.13 mmol), hydrogen was replaced, and the reaction was allowed to react at 50 ° C for 12 hours under a hydrogen atmosphere. The reaction was cooled to room temperature, filtered through diatomaceous earth, the filter cake was washed with methanol (50 mL × 3), the filtrate was collected and concentrated, and the crude product was purified by preparative high performance liquid chromatography column (chromatographic column: Waters-Xbridge-C18, 19 × 250 mm, 10 μm; mobile phase: 10 mM ammonium bicarbonate aqueous solution-acetonitrile; gradient: acetonitrile 20%-50%, 12.5 min) to obtain compound 4. MS m / z (ESI): 495.2 [M+H] + .

[0152] 1 H NMR(400MHz, DMSO-d6)δ:9.71(s,1H),8.21(s,1H),7.39(d,J=2.8Hz,1H),7.19(d,J=8.4H z,1H),7.04-6.94(m,1H),5.40-5.19(m,1H),4.28-4.19(m,1H),4.18-3.95(m,3H),3.74(s ,3H),3.14-3.04(m,2H),3.04-2.96(m,1H),2.88-2.77(m,1H),2.68-2.61(m,1H),2.22-2. 09(m,1H),2.09-2.04(m,1H),2.04-1.96(m,1H),1.96-1.62(m,4H),0.83(d,J=6.8Hz,6H).

[0153] Example 5: Preparation of Compound 5

[0154]

[0155] Step 1: Synthesis of compound 5-2

[0156]

[0157] Compound 5-1 (5.0 g, 27.5 mmol) was dissolved in tetrahydrofuran (100 mL), and lithium bis(trimethylsilyl)amide (15.2 mL, 30.3 mmol, 2 mol / L) was added at -78°C under nitrogen atmosphere, and then stirred for 1 hour, and iodoisopropyl (5.15 g, 30.3 mmol) was added, and the temperature was raised to -40°C for reaction for 1 hour. The temperature was lowered to -78°C, and added dropwise to a saturated ammonium chloride solution (100 mL), and diluted with water (50 mL), and extracted with ethyl acetate (100 mL×3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the crude product was purified by rapid silica gel column chromatography (ethyl acetate / petroleum ether) to obtain compound 5-2.

[0158] 1 HNMR(400MHz, CDCl3)δ:7.41(d,J=8.4Hz,1H),6.94(d,J=2.8Hz,1H),6.86(dd,J=8.4,2.8Hz,1H),4.13( d,J=5.6Hz,1H),3.81(s,3H),2.16(td,J=6.4,5.6Hz,1H),1.10(d,J=6.8Hz,3H),1.03(d,J=6.8Hz,3H).

[0159] Step 2: Synthesis of compound 5-3

[0160]

[0161] Compound 5-2 (3.7 g, 16.5 mmol) was dissolved in methanol (20 mL), and aqueous ammonia (1 mL) and Raney nickel aqueous solution (3.0 mL) were added to replace the hydrogen, and then stirred for 18 hours, filtered to obtain the filtrate, and the crude product was purified by a rapid silica gel column (eluent: eluent: dichloromethane / methanol) after reduced pressure concentration to obtain compound 5-3. MS m / z (ESI): 228.2 [M+H] + .

[0162] 1 HNMR(400MHz,DMSO-d6)δ:7.23(d,J=8.8Hz,1H),6.99(d,J=2.8Hz,1H),6.91(dd,J=8.8,2.8Hz,1H), 3.76(s,3H),2.91–2.75(m,3H),1.87(q,J=6.8Hz,1H),0.93(d,J=6.8Hz,3H),0.69(d,J=6.8Hz,3H).

[0163] Step 3: Synthesis of compound 5-4

[0164]

[0165] Dissolve triphosgene (1.6 g, 5.3 mmol) in dichloromethane (30 mL), add compound 5-3 (2.0 g, 8.8 mmol) and triethylamine (3.1 g, 30.7 mmol) under nitrogen atmosphere at 0°C, and stir for 1 hour. Concentrate the reaction solution under reduced pressure, add water (50 mL) to dilute, extract with ethyl acetate (100 mL×3), dry the organic phase with anhydrous sodium sulfate, filter and concentrate to obtain compound 5-4. MS m / z (ESI): 254.2 [M+H] + .

[0166] Step 4: Synthesis of compound 5-5

[0167]

[0168] Compound 5-4 (1.8 g, 7.1 mmol) was dissolved in trifluoromethanesulfonic acid (10 mL), heated to 100 °C and stirred for 1 hour. The mixture was cooled to 0 °C and added dropwise to saturated sodium bicarbonate (20 mL), diluted with water (30 mL), extracted with ethyl acetate (100 mL × 3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified using a fast silica gel column (eluent: dichloromethane / methanol) to obtain compound 5-5. MS m / z (ESI): 240.0 [M+H] + .

[0169] 1 HNMR(400MHz, Methanol-d4)δ:7.33(d,J=2.4Hz,1H),7.02(d,J=2.4Hz,1H),3.58(dd,J=13.2,1.6Hz,1H),3.48(dd,J=13.2 ,4.4Hz,1H),2.94(ddd,J=6.4,4.4,1.6Hz,1H),1.96(dd,J=13.6,6.8Hz,1H),0.98(d,J=6.8Hz,3H),0.95(d,J=6.8Hz,3H).

[0170] Step 5: Synthesis of Compound 5-6

[0171]

[0172] Compound 5-5 (105 mg, 0.44 mmol) was dissolved in acetonitrile (10 mL), potassium carbonate (121 mg, 0.88 mmol) and benzyl bromide (90 mg, 0.53 mmol) were added, and the temperature was raised to 70°C for reaction for 5 hours. The mixture was cooled to room temperature, diluted with water (20 mL), extracted with ethyl acetate (50 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 5-6. MS m / z (ESI): 330.2 [M+H] + .

[0173] 1 HNMR(400MHz, Methanol-d4)δ:7.42(d,J=2.8Hz,1H),7.30–7.25(m,2H),7.24–7.18(m,2H),7.18–7.13(m,1H),7.06(d,J=2.8Hz,1H),4.92(s,2H ), 3.43(dd,J=13.6,1.6Hz,1H), 3.30(dd,J=13.6,4.4Hz,1H), 2.80(ddd,J=6.4,4.4,1.6Hz,1H), 1.86–1.78(m,1H), 0.81(dd,J=14.0,6.8Hz,6H). 1.86–1.78(m,1H)

[0174] Step 6: Synthesis of Compound 5-7

[0175]

[0176] Compound 5-6 (144 mg, 0.44 mmol) was dissolved in tetrahydrofuran (5 mL), sodium hydride (23 mg, 0.57 mmol, purity 60%) was added at 0°C under nitrogen atmosphere, then stirred for 1 hour, 4-1 (133 mg, 0.65 mmol) was added, and the mixture was reacted at 0°C for 1 hour. The mixture was added dropwise to a saturated ammonium chloride solution (5 mL), diluted with water (10 mL), extracted with ethyl acetate (20 mL×3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified using a fast silica gel column (eluent: ethyl acetate / petroleum ether) to obtain compound 5-7. MS m / z (ESI): 496.2 [M+H] + .

[0177] Step 7: Synthesis of Compound 5-8

[0178]

[0179] Compound 5-7 (100 mg, 0.20 mmol) was dissolved in dioxane (2 mL), ((2R, 7aS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (64 mg, 0.40 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (23 mg, 0.04 mmol), cesium carbonate (196 mg, 0.60 mmol) were added, and the mixture was reacted at 130°C in a microwave reactor for 3 hours after nitrogen replacement. Water (10 mL) was added for dilution, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by rapid silica gel column chromatography (dichloromethane / methanol) to obtain compound 5-8. MS m / z(ESI):619.4[M+H] + .

[0180] Step 8: Synthesis of compound 5

[0181]

[0182] Compound 5-8 (30 mg, 0.05 mmol) was dissolved in trifluoroacetic acid (2 mL) and heated to 80°C for 7 hours. The reaction solution was decompressed to remove the solvent and purified by preparative HPLC (chromatographic column: Boston Prime C18, 150×30 mm, 5 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 15%-45%, 15 min) to obtain compound 5. MS m / z (ESI): 529.3 [M+H] + . 1 HNMR(400MHz,Methanol-d4)δ:8.24(s,1H),7.57–7.49(m,1H),7.13(d,J=2.8H z,1H),5.55(d,J=52.0Hz,1H),4.72–4.56(m,2H),4.55–4.47(m,1H),4.26–4.16 (m,1H),3.84(d,J=14.8Hz,6H),3.43(d,J=5.2Hz,1H),3.19–3.12(m,1H),2.78– 2.54(m,2H),2.44–2.08(m,5H),0.98(dd,J=6.8,2.4Hz,3H),0.96–0.92(m,3H).

[0183] Example 6: Preparation of Compound 6

[0184]

[0185] Step 1: Synthesis of compound 6-2

[0186]

[0187] At 0°C, thionyl chloride (8.76 mL, 120.83 mmol) was slowly added to a solution of compound 6-1 (3 g, 24.17 mmol) in tetrahydrofuran (40 mL), and the temperature was naturally raised to 25°C and stirred for 18 hours. A large amount of light yellow solid precipitated in the reaction solution, which was filtered out and rinsed with petroleum ether (50 mL), and the solid was dried under reduced pressure to obtain compound 6-2.

[0188] 1 H NMR (400MHz, DMSO-d6) δ: 8.29 (s, 2H), 8.11 (dd, J = 7.2, 1.6 Hz, 1H), 8.04 (dd, J = 6.4, 1.6 Hz, 1H), 6.96-6.89 (m, 1H), 4.86 (s, 2H).

[0189] Step 2: Synthesis of compound 6-3

[0190]

[0191] A solution of compound 6-2 (2.54 g, 14.18 mmol, hydrochloride), compound 6-3 (2.45 g, 14.18 mmol) and potassium carbonate (5.88 g, 42.53 mmol) in acetonitrile (40 mL) was heated to 70 ° C and stirred for 1 hour. The reaction solution was diluted with water (200 mL) and extracted with ethyl acetate (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 6-4. MS m / z (ESI): 278.6 [M + H] + . 1 H NMR (400MHz, DMSO-d6) δ: 8.73 (s, 1H), 7.92 (dd, J = 4.8, 2.0 Hz, 1H), 7.22-7.11 (m, 1H), 6.52 (dd, J = 7.2, 4.8 Hz, 1H), 6.12 (s, 2H), 5.31 (s, 2H).

[0192] Step 3: Synthesis of compound 6-5

[0193]

[0194] A mixed solution of compound 6-4 (500 mg, 1.79 mmol), di-tert-butyl dicarbonate (1.15 mL, 5.38 mmol) and 4-dimethylaminopyridine (109.60 mg, 0.9 mmol) in tetrahydrofuran (8 mL) and N,N-dimethylformamide (2 mL) was heated to 50 ° C and stirred for 3 hours. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 6-5. 1 H NMR (400MHz, DMSO-d6) δ: 8.62 (s, 1H), 8.50 (dd, J = 4.8, 2.0 Hz, 1H), 8.05 (dd, J = 7.6, 2.0 Hz, 1H), 7.51 (dd, J = 7.6, 4.8 Hz, 1H), 5.39 (s, 2H), 1.24 (s, 18H).

[0195] Step 4: Synthesis of compound 6-6

[0196]

[0197] Compound 6-5 (102.36 mg, 0.21 mmol), compound 1-6 (45 mg, 0.14 mmol), cesium carbonate (92.86 mg, 0.28 mmol), palladium acetate (4.74 mg, 0.03 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (16.49 mg, 0.03 mmol) and Molecular sieves (8 mg) were dissolved in 1,4-dioxane (1.5 mL), heated to 90°C in a nitrogen atmosphere, and stirred for 6 hours. The reaction solution was filtered with a filter membrane and purified using a silica gel preparation plate (methanol / dichloromethane) to obtain compound 6-6. MS m / z (ESI): 758.5 ​​[M+H] + .

[0198] Step 5: Synthesis of Compound 6-7

[0199]

[0200] At 25°C, a solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (22.68 mg, 0.14 mmol) in tetrahydrofuran (1 mL) was added with a solution of potassium tert-butoxide in tetrahydrofuran (142.43 μL, 0.14 mmol, 1 M), and the mixture was heated to 30°C and stirred for 10 minutes. A solution of compound 6-6 (36 mg, 0.05 mmol) in tetrahydrofuran (1 mL) was then added at 0°C, and the reaction mixture was stirred at 0°C for 1 minute. The reaction mixture was quenched with trifluoroacetic acid (0.1 mL) and concentrated under reduced pressure to obtain a crude compound 6-7. MS m / z(ESI):797.5[M+H] + .

[0201] Step 6: Synthesis of compound 6

[0202]

[0203] Compound 6-7 (45 mg, 0.06 mmol) was dissolved in trifluoroacetic acid (1 mL), and the solution was heated to 70°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure and then purified by preparative HPLC column (chromatographic column: Xtimate C18, 150×40 mm, 5 μm; mobile phase: 0.01% ammonium bicarbonate aqueous solution-acetonitrile; gradient: acetonitrile 45%-75%, 19 min) to obtain compound 6. MS m / z (ESI): 607.4 [M+H] + .

[0204] 1 H NMR(400MHz, DMSO-d6)δ:10.30(s,1H),8.36(s,1H),7.92(dd,J=5.2,2.0Hz,1H),7.39(d,J=2.8Hz,1H),7 .25(dd,J=7.6,2.0Hz,1H),7.12(d,J=2.4Hz,1H),6.54(dd,J=7.6,4.8Hz,1H),6.16(s,2H),5.27(d,J=54. 0Hz,1H),5.24(s,2H),4.24-3.97(m,4H),3.16-3.05(m,3H),3.00(s,1H),2.87-2.77(m,1H),2.21-2.10( m,1H),2.11-2.03(m,1H),2.05-1.94(m,1H),1.91-1.73(m,3H),1.71-1.53(m,2H),0.87(t,J=7.6Hz,3H).

[0205] Example 7: Preparation of Compound 7

[0206]

[0207] Step 1: Synthesis of compound 7-2

[0208]

[0209] A solution of compound 7-1 (2.90 g, 17.36 mmol, hydrochloride), compound 6-3 (3 g, 17.36 mmol) and potassium carbonate (7.20 g, 52.09 mmol) in acetonitrile (100 mL) was heated to 70°C and stirred for 18 hours. The reaction solution was filtered and concentrated under reduced pressure. The crude product was purified using a silica gel column (eluent: ethyl acetate / petroleum ether) to obtain compound 7-2. MS m / z (ESI): 267.0 [M+H] + .

[0210] 1 H NMR (400MHz, DMSO-d6) δ: 8.75 (s, 1H), 7.64 (d, J = 2.0 Hz, 1H), 6.26 (d, J = 2.0 Hz, 1H), 5.41 (s, 2H), 3.77 (s, 3H).

[0211] Step 2: Synthesis of compound 7-4

[0212]

[0213] Compound 7-3 (7.0 g, 32.55 mmol) was dissolved in tetrahydrofuran (100 mL), and cyclopropylmagnesium bromide (150 mL, 0.5 mol / L) was added at 0°C and reacted for 1 hour. Saturated ammonium chloride (100 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (200 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a filtrate. After reduced pressure concentration, the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 7-4. MS m / z (ESI): 239.2 [M-OCH3] + .

[0214] Step 3: Synthesis of compound 7-5

[0215]

[0216] Compound 7-4 (9.0 g, 35.00 mmol) was dissolved in tetrahydrofuran (70 mL), and the reaction solution was cooled to 0 ° C under nitrogen protection. Sodium hydride (2.8 g, 70.00 mmol, 60%) was added and reacted for 0.5 hours. Methyl iodide (7.45 g, 52.50 mmol) was added and reacted for 1 hour. Saturated ammonium chloride (100 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (200 mL × 3). The organic phase was dried over anhydrous sodium sulfate and filtered, and then concentrated to obtain a filtrate. After reduced pressure concentration, the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 7-5. MS m / z (ESI): 239.2 [M-OCH3] + .

[0217] Step 4: Synthesis of compound 7-6

[0218]

[0219] Compound 7-5 (7.5 g, 27.66 mmol) was dissolved in dichloromethane (100 mL), the reaction solution was cooled to 0°C, ferric chloride (0.45 g, 2.77 mmol) and trimethylsilyl cyanide (8.23 g, 82.98 mmol) were added, and the reaction was continued for 18 hours. After the reaction solution was concentrated under reduced pressure, the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 7-6. MS m / z (ESI): 239.2 [M-CN] + .

[0220] Step 5: Synthesis of compound 7-7

[0221]

[0222] Compound 7-6 (3.8 g, 14.3 mmol) was dissolved in N, N-dimethylformamide (60 mL) and methanol (20 mL), potassium acetate (3.50 g, 35.7 mmol), 1,3-bis(diphenylphosphine)propane (1.18 g, 2.86 mmol) and palladium acetate (0.32 g, 1.43 mmol), carbon monoxide gas was replaced 3 times, and the mixture was reacted for 18 hours under a carbon monoxide gas atmosphere (50 psi) at 80 °C. Water (200 mL) was added for dilution, and the mixture was extracted with ethyl acetate (100 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 7-7. MS m / z (ESI): 246.2 [M+H] + .

[0223] 1H NMR(400MHz, DMSO-d6)δ:7.64(d,J=8.8Hz,1H),7.38(d,J=2.8Hz,1H),7.33-7.22(m,1H),4.46(d,J=8.0Hz, 1H),3.86(s,3H),3.82(s,3H),1.56-1.37(m,1H),0.73-0.61(m,1H),0.59-0.43(m,2H),0.42-0.30(m,1H).

[0224] Step 6: Synthesis of compound 7-8:

[0225]

[0226] Compound 7-7 (2.6 g, 10.6 mmol) was dissolved in methanol (120 mL), 7M ammonia methanol solution (60 mL) and Raney nickel (2.32 g) were added, hydrogen was replaced 3 times, and the mixture was reacted for 18 hours at 50°C under a hydrogen atmosphere (50 psi). The reaction mixture was filtered through diatomaceous earth, the filter cake was rinsed with methanol (300 mL), and the filtrate was concentrated and purified using a fast silica gel column (eluent: ethyl acetate / petroleum ether) to obtain compound 7-8. MS m / z (ESI): 218.1 [M+H] + .

[0227] 1 H NMR(400MHz,DMSO-d6)δ:7.96(br s,1H),7.40(d,J=8.4Hz,1H),7.37(d,J=2.8Hz,1H),7.11-7.06(m,1H),3.78(s,3H),3.50-3.42(m, 1H),3.32-3.24(m,1H),2.15-2.04(m,1H),1.00-0.79(m,1H),0.61-0.42(m,2H),0.41-0.19(m,2H).

[0228] Step 7: Synthesis of Compound 7-9

[0229]

[0230] Boron tribromide (2.18 mL, 23.01 mmol) was added to a dichloromethane solution (25 mL) of compound 7-8 (1 g, 4.60 mmol) at 0°C, and the reaction solution was stirred at 0°C for 2 hours. The reaction solution was quenched with water (40 mL) and extracted with dichloromethane / methanol (10:1, v / v, 50 mL×2). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 7-9. MS m / z (ESI): 204.1 [M+H] + .

[0231] Step 8: Synthesis of Compound 7-10

[0232]

[0233] A mixed solution of compound 7-9 (820 mg, 4.03 mmol), benzyl bromide (0.531 mL, 4.44 mmol), potassium carbonate (1115.17 mg, 8.07 mmol) in acetonitrile (20 mL) and N,N-dimethylformamide (3 mL) was stirred at 50°C for 18 hours. Filter, concentrate the filtrate, and purify the crude product by rapid silica gel column chromatography (eluent: petroleum ether / ethyl acetate) to obtain compound 7-10. MSm / z(ESI):294.2[M+H] + .

[0234] 1 H NMR(400MHz,DMSO-d6)δ:7.95(br s,1H),7.48-7.43(m,3H),7.42-7.37(m,3H),7.35-7.30(m,1H),7.17-7.14(m,1H),5.14(s,2H),3.47-3.43(m,1H),3.30-3. 24(m,1H),2.12-2.07(m,1H),0.95-0.86(m,1H),0.57-0.50(m,1H),0.49-0.42(m,1H),0.39-0.32(m,1H),0.27-0.21(m,1H).

[0235] Step 9: Synthesis of Compound 7-11

[0236]

[0237] Compound 7-2 (90.90 mg, 0.34 mmol), compound 7-10 (100 mg, 0.34 mmol), cesium carbonate (222.12 mg, 0.68 mmol), palladium acetate (5.67 mg, 0.03 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (39.45 mg, 0.07 mmol) and Molecular sieves (20 mg) were dissolved in 1,4-dioxane (1.5 mL), and the mixture was bubbling with nitrogen to remove air for 2 minutes, and then replaced with a nitrogen atmosphere, heated to 115°C and stirred for 2 hours. The reaction solution was filtered with a filter membrane and purified using a silica gel preparation plate to obtain compound 7-11. MS m / z (ESI): 524.4 [M+H] + .

[0238] Step 10: Synthesis of Compound 7-12

[0239]

[0240] At 25°C, a solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (109.46 mg, 0.69 mmol) in tetrahydrofuran (1.5 mL) was added with a solution of potassium tert-butoxide in tetrahydrofuran (687.59 μL, 1 M), and the solution was heated to 30°C and stirred for 10 minutes. A solution of compound 7-11 (120 mg, 0.23 mmol) in tetrahydrofuran (1.5 mL) was then added at 0°C, and the reaction solution was stirred at 0°C for 1 minute. After the reaction was completed, the reaction solution was quenched with trifluoroacetic acid (0.1 mL) and concentrated under reduced pressure to obtain compound 7-12. MS m / z (ESI): 663.5 [M+H] + .

[0241] Step 11: Synthesis of compound 7

[0242]

[0243] Compound 7-12 (130 mg, 0.20 mmol) was dissolved in trifluoroacetic acid (2 mL), and the solution was heated to 70°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure and then purified by preparative HPLC column (chromatographic column: Xtimate C18, 150×40 mm, 5 μm; mobile phase: 0.01% ammonium bicarbonate aqueous solution-acetonitrile; gradient: acetonitrile 30%-95%, 15.7 min) to obtain compound 7. MS m / z (ESI): 573.3 [M+H] + .

[0244] 1H NMR(400MHz, DMSO-d6)δ:9.72(s,1H),8.28(s,1H),7.64(d,J=2.0Hz,1H),7.45-7.38(m,2H),7.03(dd,J =8.4,2.7Hz,1H),6.26(d,J=2.0Hz,1H),5.32(s,2H),5.40-5.21(m,1H),4.24-4.13(m,2H),4.11-4.02(m ,2H),3.78(s,3H),3.20-2.99(m,3H),2.91-2.80(m,1H),2.37-2.29(m,1H),2.21-2.09(m,2H),2.06-1.9 8(m,1H),1.94-1.73(m,3H),1.06-0.94(m,1H),0.60-0.50(m,1H),0.49-0.34(m,2H),0.30-0.20(m,1H).

[0245] Example 8: Preparation of Compound 8

[0246]

[0247] Step 1: Synthesis of compound 8-1

[0248]

[0249] Compound 6-5 (71.83 mg, 0.15 mmol), compound 7-10 (55 mg, 0.19 mmol), cesium carbonate (122.17 mg, 0.37 mmol), palladium acetate (3.12 mg, 0.02 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (21.70 mg, 0.04 mmol) and Molecular sieves (10 mg) were dissolved in 1,4-dioxane (1 mL), heated to 115°C in a nitrogen atmosphere, and stirred for 2 hours. The reaction solution was filtered with a filter membrane and purified using a silica gel preparation plate (methanol / dichloromethane) to obtain compound 8-1. MS m / z (ESI): 736.5 [M+H] + .

[0250] Step 2: Synthesis of compound 8-2

[0251]

[0252] At 25°C, a solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (14.93 mg, 0.09 mmol) in tetrahydrofuran (0.5 mL) was added with a solution of potassium tert-butoxide in tetrahydrofuran (93.77 μL, 0.09 mmol, 1 M), and the solution was heated to 30°C and stirred for 10 minutes. A solution of compound 8-1 (23 mg, 0.03 mmol) in tetrahydrofuran (0.5 mL) was then added at 0°C, and the reaction solution was stirred at 0°C for 1 minute. The reaction solution was quenched with trifluoroacetic acid (0.1 mL) and concentrated under reduced pressure to obtain a crude compound 8-2. MS m / z(ESI):775.5[M+H] + .

[0253] Step 3: Synthesis of compound 8

[0254]

[0255] Compound 8-2 (24 mg, 0.03 mmol) was dissolved in trifluoroacetic acid (1 mL), and the solution was heated to 70°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure and then purified by preparative HPLC column (chromatographic column: Xtimate C18, 150×40 mm, 5 μm; mobile phase: 0.025% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 32%-95%, 15.7 min) to obtain compound 8. MS m / z (ESI): 585.3 [M+H] + .

[0256] 1H NMR(400MHz, DMSO-d6)δ:9.72(s,1H),8.34(s,1H),7.91(dd,J=5.2,1.8Hz,1H),7.45-7.39(m,2H),7.23(dd,J=7.2,1.8Hz,1H),7. 03(dd,J=8.4,2.7Hz,1H),6.53(dd,J=7.2,4.8Hz,1H),6.16(s,2H),5.37-5.19(m,1H),5.24(s,2H),4.24-4.16(m,1H),4.09-4.02 (m,2H),3.11-3.07(m,1H),2.88-2.79(m,1H),2.18-2.11(m,1H),2.10-2.04(m,2H),2.03-1.97(m,1H),1.88-1.76(m,3H),1.25-1 .19(m,1H),1.14(d,J=6.8Hz,1H),1.03-0.95(m,1H),0.92-0.82(m,1H),0.59-0.51(m,1H),0.47-0.35(m,2H),0.29-0.21(m,1H).

[0257] Example 9: Preparation of Compound 9

[0258]

[0259] Step 1: Synthesis of compound 9-1

[0260]

[0261] Compound 7-2 (72.77 mg, 0.27 mmol), compound 5-6 (60 mg, 0.18 mmol), cesium carbonate (118.54 mg, 0.36 mmol), palladium acetate (3.03 mg, 0.02 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (21.05 mg, 0.04 mmol) and Molecular sieves (10 mg) were dissolved in 1,4-dioxane (1.5 mL), and the mixture was bubbling with nitrogen to remove air for 2 minutes, and then replaced with a nitrogen atmosphere, and heated to 115°C and stirred for 3 hours. The reaction solution was filtered with a filter membrane and purified using a silica gel preparation plate to obtain compound 9-1. MS m / z (ESI): 560.4 [M+H] + .

[0262] Step 2: Synthesis of compound 9-2

[0263]

[0264] At 25°C, a solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (46.90 mg, 0.29 mmol) in tetrahydrofuran (1 mL) was added with a solution of potassium tert-butoxide in tetrahydrofuran (294.63 μL, 1 M), and the solution was heated to 30°C and stirred for 10 minutes. A solution of compound 9-1 (55 mg, 0.10 mmol) in tetrahydrofuran (1.5 mL) was then added at 0°C, and the reaction solution was stirred at 0°C for 1 minute. The reaction solution was quenched with trifluoroacetic acid (0.1 mL) and concentrated under reduced pressure to obtain compound 9-2. MS m / z (ESI): 699.3 [M+H] + .

[0265] Step 3: Synthesis of compound 9

[0266]

[0267] Compound 9-2 (69 mg, 0.10 mmol) was dissolved in trifluoroacetic acid (1 mL), and the solution was heated to 70°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure and then purified by preparative HPLC column (chromatographic column: Xtimate C18, 150×40 mm, 5 μm; mobile phase: 0.025% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 15%-35%, 19 min) to obtain compound 9. MS m / z (ESI): 609.2 [M+H] + .

[0268] 1 H NMR (400MHz, DMSO-d6) δ: 10.41 (s, 1H), 8.30 (s, 1H), 7.65 (d, J = 2.2Hz, 1H), 7.41 (d, J = 2.6Hz, 1H), 7. 13(d,J=2.6Hz,1H),6.27(d,J=2.2Hz,1H),5.33(s,2H),5.28(d,J=53.2Hz,1H),4.27(d,J=13.5Hz,1 H),4.29–4.02(m,3H),3.79(s,3H),3.12–3.02(m,3H),3.01(s,1H),2.88–2.78(m,1H),2.19–2.11(m ,1H),2.04(dt,J=18.8,5.0Hz,3H),1.89–1.72(m,3H),0.91(d,J=6.9Hz,3H),0.86(d,J=6.8Hz,3H).

[0269] Example 10: Preparation of Compound 10

[0270]

[0271] Step 1: Synthesis of compound 10-1

[0272]

[0273] Compound 6-5 (104.55 mg, 0.22 mmol), compound 5-6 (60 mg, 0.18 mmol), cesium carbonate (118.54 mg, 0.36 mmol), palladium acetate (3.03 mg, 0.02 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (21.05 mg, 0.04 mmol) and Molecular sieves (10 mg) were dissolved in 1,4-dioxane (1 mL), and the mixture was bubbling with nitrogen to remove air for 2 minutes, and then replaced with a nitrogen atmosphere, and heated to 115°C and stirred for 3 hours. The reaction solution was filtered with a filter membrane and purified using a silica gel preparation plate to obtain compound 10-1. MS m / z (ESI): 772.3 [M+H] + .

[0274] Step 2: Synthesis of compound 10-2

[0275]

[0276] At 25°C, a solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (35.25 mg, 0.22 mmol) in tetrahydrofuran (1.5 mL) was added with a solution of potassium tert-butoxide in tetrahydrofuran (210 μL, 0.21 mmol), and the solution was heated to 30°C and stirred for 10 minutes. A solution of compound 10-1 (57 mg, 0.07 mmol) in tetrahydrofuran (1 mL) was then added at 0°C, and the reaction solution was stirred at 0°C for 1 minute. The reaction solution was quenched with trifluoroacetic acid (0.1 mL) and concentrated under reduced pressure to obtain a crude compound 10-2. MS m / z(ESI):811.4[M+H] + .

[0277] Step 3: Synthesis of compound 10

[0278]

[0279] Compound 10-2 (59 mg, 0.07 mmol) was dissolved in trifluoroacetic acid (1 mL), and the solution was heated to 70°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure and then chromatographed using a preparative HPLC column (chromatographic column: Xtimate C18, 150×40 mm, 5 μm; mobile phase: 0.025% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 10%-30%, 19 min) to obtain compound 10 (formate). MS m / z (ESI): 621.0 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ: 10.23 (s, 1H), 8.39 (s, 1H), 7.95-7.89 (m, 1H), 7.42 (d, J = 2.8Hz, 1H), 7.23 (d ,J=7.2Hz,1H),7.13(d,J=2.8Hz,1H),6.54(dd,J=7.6,4.9Hz,1H),6.18(s,2H),5.45-5.26(m,1H),5. 24(s,2H),4.33-4.07(m,4H),3.26-3.15(m,3H),3.10-3.02(m,1H),2.92(s,1H),2.29-2.19(m,1H),2 .19-2.12(m,1H),2.12-2.00(m,2H),1.96-1.77(m,3H),0.91(d,J=6.8Hz,3H),0.87(d,J=6.8Hz,3H).

[0280] Example 11: Preparation of Compound 11, Compound 11-P1, Compound 11-P2

[0281]

[0282] Step 1: Synthesis of compound 11-1

[0283]

[0284] Compound 6-2 (19 g, 106.12 mmol, hydrochloride) was heated to 100°C in acetic anhydride (150 mL) and reacted for 24 hours. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 11-1. MS m / z (ESI): 227.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ: 8.62–8.56 (m, 1H), 8.16–8.11 (m, 1H), 7.60 (dd, J = 8.0, 4.0Hz, 1H), 4.69 (s, 2H), 2.20 (s, 6H).

[0285] Step 2: Synthesis of compound 11-3

[0286]

[0287] At 0°C, sodium hydride (928 mg, 23.20 mmol, purity 60%) was slowly added to a solution of compound 11-2 (3.86 g, 19.33 mmol) in N, N-dimethylformamide (80 mL), followed by compound 11-1 (4.38 g, 19.33 mmol). The reaction solution was naturally heated to 25°C and stirred for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (100 mL) and diluted with water (500 mL), and then extracted with ethyl acetate (50 mL×3). The combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was purified using a silica gel column (eluent: ethyl acetate / petroleum ether) to obtain compound 11-3. MS m / z(ESI):348.1[M+H] + .

[0288] Step 3: Synthesis of compound 11-4

[0289]

[0290] Compound 11-3 (370 mg, 1.06 mmol), compound 1-6 (280 mg, 0.89 mmol), cesium carbonate (866.65 mg, 2.66 mmol), palladium acetate (25 mg, 0.15 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (102.61 mg, 0.18 mmol) and Molecular sieves (220 mg) were dissolved in 1,4-dioxane (6 mL), and the mixed solution was bubbling with nitrogen to remove air for 2 minutes, and then replaced with a nitrogen atmosphere, heated to 115 ° C and stirred for 4 hours. The reaction solution was filtered through a filter membrane and concentrated, and the obtained crude product was purified using a silica gel column (eluent: ethyl acetate / petroleum ether) to obtain compound 11-4. MS m / z (ESI): 627.3 [M+H] + .

[0291] 1H NMR(400MHz, DMSO-d6)δ:10.32(s,1H),8.35(dd,J=4.8,2.0Hz,1H),7.64-7.61(m,1H), 7.51-7.45(m,3H),7.44-7.39(m,2H),7.38-7.32(m,3H),7.26-7.21(m,1H),6.32(d,J= 3.6Hz,1H),5.35(s,2H),5.23(s,2H),4.43-4.33(m,1H),4.13(dd,J=13.2,3.6Hz,1H), 3.25-3.16(m,1H),2.53(s,3H),2.11(s,3H),1.67-1.56(m,2H),0.89(t,J=7.6Hz,3H).

[0292] Step 4: Synthesis of compound 11-5

[0293]

[0294] A mixture of compound 11-4 (30 mg, 0.05 mmol) and potassium persulfate complex salt (18.44 mg, 0.05 mmol) in tetrahydrofuran (1 mL) and water (0.2 mL) was stirred at 25°C for 18 hours. The reaction solution was diluted with water (3 mL) and extracted with ethyl acetate (2 mL×3). The combined organic phases were washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude compound 11-5. MS m / z (ESI): 643.3 [M+H] + .

[0295] Step 5: Synthesis of compound 11-6

[0296]

[0297] At 25°C, a solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (121.78 mg, 0.76 mmol) in tetrahydrofuran (1 mL) was added with a solution of potassium tert-butoxide in tetrahydrofuran (765 μL, 1 M), and the mixture was heated to 30°C and stirred for 30 minutes. A solution of compound 11-5 (164 mg, 0.25 mmol) in tetrahydrofuran (2 mL) was then added at 0°C, and the reaction mixture was stirred at 0°C for 1 minute. The reaction mixture was quenched with trifluoroacetic acid (0.1 mL) and concentrated under reduced pressure to obtain a crude compound 11-6. MS m / z(ESI):738.5[M+H] + .

[0298] Step 6: Synthesis of compound 11-7

[0299]

[0300] Compound 11-6 (170 mg, 0.23 mmol) was dissolved in trifluoroacetic acid (3 mL), and the solution was heated to 70°C and stirred for 18 hours. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified using a silica gel column (eluent: methanol / dichloromethane) to obtain compound 11-7. MS m / z (ESI): 648.3 [M+H] + .

[0301] Step 7: Synthesis of Compound 11-P1 and Compound 11-P2

[0302]

[0303] Compound 11-7 (150 mg, 0.23 mmol) and sodium carbonate (122.65 mg, 1.16 mmol) were added to methanol (20 mL), and the mixture was heated to 70°C and stirred for 12 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The crude product was separated by preparative supercritical fluid chromatography (chromatographic column: DAICEL CHIRALPAK AD (250×30 mm, 10 μm), mobile phase: carbon dioxide-methanol (0.1% ammonia water); gradient: methanol 40%, flow rate: 130 mL / min), and compound 11-P1 (retention time 1.192 min) and compound 11-P2 (retention time 5.937 min) were separated.

[0304] Compound 11-P1:

[0305] MS m / z(ESI):606.3[M+H] + . 1 H NMR(400MHz,DMSO-d6)δ:10.47(s,1H),7.92-7.85(m,1H),7.44-7.39(m,1H),7.35(d,J=4.0Hz,1H) ,7.13-7.08(m,1H),7.08-7.03(m,1H),6.50(dd,J=8.0,4.0Hz,1H),6.27(d,J=4.0Hz,1H),6.11(s, 2H),5.37-5.13(m,3H),4.40-4.31(m,1H),4.18-3.96(m,3H),3.16-2.99(m,4H),2.87-2.78(m,1H) ,2.17-2.10(m,1H),2.08-1.95(m,2H),1.88-1.71(m,3H),1.62-1.52(m,2H),0.86(t,J=8.0Hz,3H).

[0306] Compound 11-P2:

[0307] MS m / z(ESI):606.3[M+H] + . 1 H NMR(400MHz, DMSO-d6)δ:10.19(s,1H),7.91-7.86(m,1H),7.44(d,J=4.0Hz,1H),7.36(d,J=4.0Hz,1 H),7.13(d,J=4.0Hz,1H),7.08-7.04(m,1H),6.50(dd,J=8.0,4.0Hz,1H),6.28(d,J=4.0Hz,1H),6.11 (s,2H),5.37-5.16(m,3H),4.38-4.31(m,1H),4.14-4.02(m,3H),3.20-2.97(m,4H),2.89-2.78(m,1H ),2.18-2.11(m,1H),2.09-1.98(m,2H),1.89-1.72(m,3H),1.64-1.53(m,2H),0.87(t,J=8.0Hz,3H).

[0308] Example 12: Preparation of Compound 12, Compound 12-P1, Compound 12-P2

[0309]

[0310] Step 1: Synthesis of compound 12-2

[0311]

[0312] Compound 12-1 (10 g, 73.0 mmol) was dissolved in ethanol (100 mL), sodium borohydride (2.85 g, 75.0 mmol) was added at 0 ° C, the mixture was warmed to room temperature and stirred for 3 hours. After the reaction was completed, the reaction solution was concentrated and poured into 100 mL of saturated ammonium chloride for quenching, and extracted with ethyl acetate (100 mL × 3). The organic phases were combined, washed with saturated brine (70 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain compound 12-2. MS m / z (ESI): 139.1 [M + H] + .

[0313] 1H NMR (400MHz, DMSO-d6) δ: 7.82 (dd, J=4.8, 2.0Hz, 1H), 7.39 (dd, J=7.2, 2.0Hz, 1H), 6.53 (dd, J=7.2, 4.8Hz, 1H), 5.70 (s, 2H), 5.25 (d, J = 4.0Hz, 1H), 4.71 (tt, J = 9.2, 4.4Hz, 1H), 1.31 (d, J = 6.4Hz, 3H).

[0314] Step 2: Synthesis of compound 12-3

[0315]

[0316] Compound 12-2 (9.5 g, 68.8 mmol) was dissolved in tetrahydrofuran (100 mL), thionyl chloride (16.4 g, 137.7 mmol) was added at 0°C, the temperature was raised to 25°C, and stirred for 18 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure, and toluene (100 mL×3) was added and concentrated under reduced pressure to obtain compound 12-3.

[0317] 1 H NMR(400MHz, DMSO-d6)δ:8.28(s,2H),8.22(dd,J=7.6,1.6Hz,1H),8.05(dd,J=6.4,1 .6Hz, 1H), 6.96 (dd, J=7.6, 6.4Hz, 1H), 5.52 (q, J=6.8Hz, 1H), 1.84 (d, J=6.8Hz, 3H).

[0318] Step 3: Synthesis of compound 12-4

[0319]

[0320] A solution of compound 12-3 (5.0 g, 25.90 mmol, hydrochloride), compound 6-3 (4.47 g, 25.90 mmol) and potassium carbonate (10.74 g, 77.69 mmol) in acetonitrile (80 mL) was heated to 70 ° C and stirred for 1 hour. The reaction solution was diluted with water (200 mL) and extracted with ethyl acetate (80 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 12-4. MS m / z (ESI): 293.0 [M + H] + .

[0321] Step 4: Synthesis of compound 12-5

[0322]

[0323] A tetrahydrofuran (10 mL) solution of compound 12-4 (680 mg, 2.32 mmol), di-tert-butyl dicarbonate (1.49 mL, 6.97 mmol) and 4-dimethylaminopyridine (141.91 mg, 1.16 mmol) was heated to 50 ° C and stirred for 3 hours. The reaction solution was diluted with water (50 mL) and extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, and the crude product was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 12-5. MS m / z (ESI): 515.2 [M+Na] + .

[0324] 1 HNMR(400MHz,DMSO-d6)δ:8.73(s,1H),8.51(dd,J=4.8,1.6Hz,1H),8.38(dd,J=7.6,1.6Hz,1H),7 .61(dd,J=7.6,4.8Hz,1H),5.86(q,J=7.2Hz,1H),1.94(d,J=7.2Hz,3H),1.46(s,9H),0.92(s,9H).

[0325] Step 5: Synthesis of compound 12-6

[0326]

[0327] Potassium carbonate (3.06 g, 22.16 mmol) and benzyl bromide (1.99 g, 11.63 mmol) were added to a solution of compound 1-5 (2.5 g, 11.08 mmol) in acetonitrile (50 mL) and N,N-dimethylformamide (25 mL) at 25 °C. The reaction mixture was then stirred at 50 °C for 12 hours. The mixture was poured into 100 ml of ice water and extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by fast silica gel column (eluent: ethyl acetate / petroleum ether), and then separated by supercritical fluid chromatography (chromatographic column: DAICEL CHIRALPAKAD (50×4.6mm, 3mm), mobile phase carbon dioxide-methanol (0.05% diethylamine); gradient: methanol 40%, flow rate: 3mL / min) to obtain compound 12-6 (retention time 1.183min), (Note: the retention time of the other stereoisomer is 1.644min). 1H NMR(400MHz, DMSO-d6)δ:7.99(d,J=4.8Hz,1H),7.48-7.37(m,5H),7.37-7.34(m,1H),7.33(d,J=2.8Hz,1H),5.17(s,2H),3.50-3.43(m,1H),3.34

[0328] -3.29(m,1H),2.95(td,J=4.0,8.8Hz,1H),1.61-1.39(m,2H),0.94(t,J=7.2Hz,3H).

[0329] Step 6: Synthesis of compound 12-7

[0330]

[0331] Compound 12-5 (139 mg, 0.28 mmol), compound 12-6 (89 mg, 0.28 mmol), cesium carbonate (275.63 mg, 0.85 mmol), palladium acetate (9.39 mg, 0.06 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (32.63 mg, 0.06 mmol) and Molecular sieves (8 mg) were dissolved in 1,4-dioxane (2 mL), heated to 90°C in a nitrogen atmosphere, and stirred for 6 hours. The reaction solution was filtered with a filter membrane and purified using a silica gel preparation plate (eluent: methanol / dichloromethane) to obtain compound 12-7. MS m / z (ESI): 772.4 [M+H] + .

[0332] Step 7: Synthesis of compound 12-8

[0333]

[0334] At 25°C, a solution of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolazine-7a(5H)-yl)methanol (81.19 mg, 0.17 mmol) in tetrahydrofuran (1 mL) was added with a solution of potassium tert-butoxide in tetrahydrofuran (680 μL, 0.68 mmol, 1 M), and the mixture was heated to 30°C and stirred for 10 minutes. A solution of compound 12-7 (135 mg, 0.17 mmol) in tetrahydrofuran (1 mL) was then added at 0°C, and the reaction mixture was stirred at 0°C for 1 minute. The reaction mixture was quenched with trifluoroacetic acid (0.1 mL) and concentrated under reduced pressure to obtain a crude compound 12-8.

[0335] 1HNMR (400MHz, DMSO-d6) δ: 10.72 (s, 1H), 9.34 (d, J = 6.4Hz, 1H), 8.45 (d, J = 9.2Hz, 1H), 8.41-8.36 (m, 1H), 8.16-8. 07(m,1H),7.58-7.54(m,1H),7.51-7.45(m,3H),7.41(t,J=7.2Hz,2H),7.38-7.32(m,2H),6.07-5.97(m,1H),5.6 9-5.51(m,1H),5.22(s,2H),4.56-4.43(m,2H),4.22-4.11(m,2H),3.93-3.71(m,3H),3.22-3.18(m,1H),2.39-2. 27(m,2H),2.26-2.16(m,2H),2.11-1.94(m,6H),1.70-1.55(m,2H),1.41(d,J=7.2Hz,9H),0.86(t,J=7.2Hz,3H).

[0336] Step 7: Synthesis of Compound 12-P1 and Compound 12-P2

[0337]

[0338] Compound 12-8 (45 mg, 0.06 mmol) was dissolved in trifluoroacetic acid (1 mL), and the solution was heated to 70°C and stirred for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was separated by preparative supercritical fluid chromatography (chromatographic column: ChiralPak AD, 250×30 mm ID, 10 μm, mobile phase: carbon dioxide-isopropanol; gradient: isopropanol 35%, flow rate: 150 mL / min), and compound 12-P1 (retention time 1.161 min) and compound 12-P2 (retention time 1.867 min) were separated.

[0339] Compound 12-P1:

[0340] .MS m / z(ESI):621.3[M+H] + . 1HNMR(400MHz,DMSO-d6)δ:8.48(s,1H),7.93-7.88(m,1H),7.46-7.38(m,1H),7.33-7.27(m,1H),7.05-6 .98(m,1H),6.57(dd,J=7.6,4.8Hz,1H),6.06(s,2H),5.86(q,J=7.2Hz,1H),5.35-5.17(m,1H),4.20-4. 09(m,2H),4.08-4.00(m,2H),3.11-3.04(m,3H),3.01-2.98(m,1H),2.84-2.78(m,1H),2.13-2.08(m,1H ),2.05-1.97(m,2H),1.89(d,3H),1.81-1.73(m,2H),1.61(d,J=14.8,7.6Hz,3H),0.86(t,J=7.6Hz,3H).

[0341] Compound 12-P2:

[0342] MS m / z(ESI):621.5[M+H] + . 1 HNMR(400MHz,DMSO-d6)δ:10.23(s,1H),8.47(s,1H),7.94-7.87(m,1H),7.51-7.44(m,1H),7.43-7.37(m, 1H),7.15-7.11(m,1H),6.57(dd,J=7.6,4.8Hz,1H),6.05(s,2H),5.87(q,J=6.8Hz,1H),5.36-5.17(m,1H), 4.20-4.11(m,2H),4.11-3.99(m,2H),3.16-3.04(m,3H),3.02-2.95(m,1H),2.86-2.78(m,1H),2.15-2.10 (m,1H),2.07-1.96(m,2H),1.94-1.87(m,3H),1.84-1.71(m,3H),1.68-1.52(m,2H),0.87(t,J=7.2Hz,3H).

[0343] Example 13: Preparation of Compound 13, Compound 13-P1, and Compound 13-P2

[0344]

[0345] Step 1: Synthesis of compound 13-1

[0346]

[0347] At room temperature, compound 12-6 (150 mg, 0.47 mmol) and compound 11-3 (171.87 mg, 0.47 mmol) were dissolved in 1,4-dioxane (5 mL), and palladium acetate (10.66 mg, 0.05 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthene (54.97 mg, 0.09 mmol), cesium carbonate (464.28 mg, 1.42 mmol) and molecular sieves (200 mg, 0.6 mmol) were added. The reaction solution was stirred at 115 ° C for 12 hours under a nitrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated. The crude product was purified using a fast silica gel column (eluent: ethyl acetate / petroleum ether) to obtain compound 13-1.

[0348] 1 H NMR(400MHz, CDCl3)δ:9.64-9.29(m,1H),8.48-8.42(m,1H),7.88-7.81(m,1H),7.74(dd,J=2.8,8.4Hz ,1H),7.45-7.31(m,5H),7.24(dd,J=1.2,2.4Hz,1H),7.20-7.13(m,2H),6.42(dd,J=2.4,3.6Hz,1H),6. 27-6.16(m,1H),5.17-5.05(m,2H),4.57-4.42(m,1H),4.21-4.06(m,1H),3.28-3.16(m,1H),2.64(d,J =1.6Hz, 3H), 2.42 (d, J = 13.6Hz, 3H), 1.94 (dd, J = 5.2, 7.2Hz, 3H), 1.75-1.66 (m, 2H), 1.01-0.90 (m, 3H).

[0349] Step 2: Synthesis of compound 13-2

[0350]

[0351] At room temperature, compound 13-1 (170 mg, 0.27 mmol) was dissolved in tetrahydrofuran (2.5 mL) and water (2.5 mL), potassium persulfate complex salt (89.65 mg, 0.15 mmol) was added, and stirred for 1 hour. The mixture was poured into 10 mL of water and extracted with ethyl acetate (10 mL × 2). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude compound 13-2 was obtained.

[0352] 1H NMR(400MHz, CDCl3)δ:8.49-8.40(m,1H),7.83-7.70(m,2H),7.68-7.56(m,1H),7 .46-7.34(m,5H),7.19(m,1H),6.63-6.54(m,1H),6.45-6.24(m,1H),5.16-5.08( m,2H),4.58-4.37(m,1H),4.17-4.05(m,2H),3.26-3.15(m,1H),3.00-2.74(m,3H ), 2.50-2.40 (m, 3H), 2.07-1.99 (m, 4H), 1.26 (t, J = 7.2Hz, 2H), 0.97-0.86 (m, 3H).

[0353] Step 3: Synthesis of compound 13-3

[0354]

[0355] Sodium tert-butoxide (54.84 mg, 0.57 mmol) was added to a tetrahydrofuran solution (5 mL) of compound 13-2 (150 mg, 0.23 mmol) and ((2R, 7aS)-2-fluorotetrahydro-1H-pyrrolizine-7a(5H)-yl)methanol (72.67 mg, 0.46 mmol) and stirred for one hour. The reaction mixture was poured into 5 mL of water and extracted with ethyl acetate (5 mL×2). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude compound 13-3 was obtained. MS m / z (ESI): 752.4

[0356] [M+H] + .

[0357] Step 4: Synthesis of compound 13-4

[0358]

[0359] Compound 13-3 (170 mg, 0.23 mmol) was dissolved in trifluoroacetic acid (2 mL) and stirred at 80 °C for 12 hours. The reaction mixture was concentrated. The crude product was purified using a rapid silica gel column (eluent: methanol / dichloromethane) to obtain compound 13-4. MS m / z (ESI): 662.4 [M+H] + .

[0360] Step 6: Synthesis of Compound 13-P1 and Compound 13-P2

[0361]

[0362] At room temperature, sodium carbonate (80.03 mg, 0.76 mmol) was added to a solution of compound 13-4 (100 mg, 0.15 mmol) in methanol (3 mL). Then stirred at 70 ° C for 12 hours. The reaction solution was filtered and the filtrate was concentrated to obtain a crude product. The crude product (90 mg, 0.15 mmol) was separated by preparative supercritical fluid chromatography (chromatographic column: DAICEL CHIRALPAK AD (100×4.6 mm, 5 mm), mobile phase: carbon dioxide-isopropanol (0.05% diethylamine); gradient: isopropanol 40%, flow rate: 2.5 mL / min), and compound 13-P1 (retention time 2.518 min) and compound 13-P2 (retention time 3.176 min) were separated.

[0363] Compound 13-P1:

[0364] MS m / z(ESI):620.3[M+H] + . 1 H NMR(400MHz,DMSO-d6)δ:7.87(dd,J=1.6,4.8Hz,

[0365] 1H),7.48-7.39(m,2H),7.37-7.28(m,1H),7.04-6.95(m,1H),6.59-6.51(m,1H),6.26(d,J=3.6Hz,1H), 6.02-5.86(m,3H),5.37-5.16(m,1H),4.34-4.24(m,1H),4.13-3.99(m,3H),3.13-3.04(m,3H),3.00(br s,1H),2.86-2.79(m,1H),2.19-2.11(m,1H),2.06(brd,J=2.4Hz,1H),2.03-1.9 8(m,1H),1.88-1.71(m,6H),1.56(td,J=7.2,14.4Hz,2H),0.84(t,J=7.6Hz,3H).

[0366] Compound 13-P2:

[0367] MS m / z(ESI):620.3[M+H] + . 1H NMR(400MHz, DMSO-d6)δ:10.36-10.09(m,1H),7.88(dd,J=1.6,4.8Hz,1H),7.49(d,J=3.6Hz,1H),7.43( d,J=2.4Hz,1H),7.37(dd,J=1.2,7.6Hz,1H),7.13(d,J=2.4Hz,1H),6.56(dd,J=4.8,7.6Hz,1H),6.28(d, J=3.6Hz,1H),6.09-5.88(m,3H),5.38-5.14(m,1H),4.38-4.27(m,1H),4.16-4.02(m,3H),3.17-2.98(m ,4H),2.87-2.76(m,1H),2.13-1.97(m,3H),1.88-1.72(m,6H),1.65-1.51(m,2H),0.87(t,J=7.6Hz,3H).

[0368] Example 14: Preparation of Compound 14, Compound 14-P1, Compound 14-P2

[0369]

[0370] Step 1: Synthesis of compound 14-2

[0371]

[0372] Compound 14-1 (2.10 g, 9.25 mmol) was added to a solution of ((2R, 7aS)-2-fluorotetrahydro-1H-pyrrolazin-7a(5H)-yl)methanol (1.88 mL, 13.21 mmol) and sodium hydride (690 mg, 17.18 mmol, purity 60%) in tetrahydrofuran (40 mL) at -70°C under nitrogen atmosphere. The reaction solution was stirred at -70°C for 1 hour. Saturated ammonium chloride solution (20 mL) was slowly added to the reaction solution to quench, and then extracted with ethyl acetate (20 mL×2). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether) to obtain compound 14-2. MS m / z(ESI):306.0[M+H] + .

[0373] Step 2: Synthesis of compound 14-3

[0374]

[0375] A solution of compound 14-2 (1 g, 3.27 mmol), (R)-3-hydroxypiperidine hydrochloride (449.98 mg, 3.27 mmol) and triethylamine (1.81 mL, 13.08 mmol) in ethanol (12 mL) was heated to 50°C and stirred for 2 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by pulping (ethyl acetate / petroleum ether) to obtain compound 14-3. MS m / z (ESI): 371.2 [M+H] + .

[0376] Step 3: Synthesis of compound 14

[0377]

[0378] Compound 3-5 (100 mg, 0.49 mmol) was dissolved in dioxane (2 mL), 14-3 (181 mg, 0.49 mmol), palladium acetate (5 mg, 0.02 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (20 mg, 0.03 mmol), cesium carbonate (480 mg, 1.47 mmol) were added, and the mixture was heated to 110°C in a nitrogen atmosphere for 2 hours. Water (10 mL) was added for dilution, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and purified by preparative high performance liquid chromatography (chromatographic column: Waters-SunFire-C18, 19×250 mm, 10 μm; mobile phase: 0.1% formic acid aqueous solution-acetonitrile, gradient: acetonitrile 20%-95%, 15 min) to obtain compound 14. MS m / z(ESI):540.4[M+H] + .

[0379] Step 4: Synthesis of Example 14-P1 and Compound 14-P2

[0380]

[0381] Example 14 (80 mg, 0.15 mmol) was separated by preparative supercritical fluid chromatography (chromatographic column: ChiralPakAD, 250×30 mm ID, 10 μm, mobile phase: carbon dioxide-ethanol (0.1% ammonia water); gradient: ethanol 25%, flow rate: 150 mL / min) to give crude products 14-P1 (retention time 3.052 min) and 14-P2 (retention time 3.648 min). The crude products 14-P1 and 14-P2 were purified by preparative high performance liquid chromatography (chromatographic column: Waters-SunFire-C18, 19×250 mm, 10 μm; mobile phase: 0.1% formic acid aqueous solution-acetonitrile, gradient: acetonitrile 20%-95%, 15 min) to give compounds 14-P1 and 14-P2.

[0382] Compound 14-P1:

[0383] MS m / z(ESI):540.4[M+H] + . 1 HNMR(400MHz,Methanol-d4)δ:7.52-7.41(m,1H),7.14(d,J=8.4Hz,1H),7.07(s,1H) ,7.02-6.92(m,1H),5.39(d,J=53.2Hz,1H),4.72(d,J=13.2Hz,1H),4.35(d,J=11.2H z,1H),4.24(d,J=11.2Hz,1H),4.16(d,J=12.8Hz,1H),3.93(d,J=13.2Hz,1H),3.83( dd,J=13.2,3.6Hz,1H),3.73-3.63(m,1H),3.60-3.42(m,3H),3.22-3.10(m,3H),2.61 -2.53(m,1H),2.51-2.30(m,2H),2.27-2.18(m,1H),2.17-2.07(m,2H),2. 06-1.93(m,2H),1.90-1.76(m,2H),1.63-1.50(m,2H),1.02-0.82(m,6H).

[0384] Compound 14-P2 (formate salt):

[0385] MS m / z(ESI):540.4[M+H] + . 1 HNMR(400MHz,Methanol-d4)δ:8.47(s,1H),7.46(d,J=

[0386] 2.4Hz,1H),7.15(d,J=8.4Hz,1H),7.06(s,1H),6.98(dd,J=8.4,2.4Hz,1H),5.48(d,J=52.4Hz,1H), 4.67(dd,J=13.2,2.4Hz,1H),4.42(s,2H),4.12(d,J=12.8Hz,1H),3.92(d,J=13.2Hz,1H),3.86(dd,J =13.2,3.6Hz,1H),3.82-3.63(m,4H),3.39-3.33(m,2H),3.20(dd,J=13.2,8.0Hz,1H),2.67-2.40(m ,3H),2.35-2.17(m,3H),2.13-1.98(m,2H),1.92-1.79(m,2H),1.66-1.49(m,2H),1.01-0.86(m,6H).

[0387] Example 15: Preparation of Compound 15

[0388]

[0389] Step 1: Synthesis of compound 15

[0390]

[0391] Compound 1-5 (100 mg, 0.49 mmol) was dissolved in dioxane (2 mL), 14-3 (246.5 mg, 0.66 mmol), palladium acetate (10 mg, 0.04 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (51.3 mg, 0.09 mmol), cesium carbonate (443.1 mg, 1.33 mmol) were added, and the temperature was raised to 110°C in a nitrogen atmosphere for 2 hours. Water (10 mL) was added for dilution, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was purified by preparative high performance liquid chromatography (chromatographic column: Boston Prime C18, 150×30 mm, 5 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 15%-45%, 17 min) to obtain compound 15 (formate salt). MS m / z(ESI):560.4[M+H] + .

[0392] 1H NMR (400MHz, DMSO-d6)δ:7.38(d,J=2.4Hz,1H),7.23(s,1H),7.08(d,J=2.4Hz,1H),5.41-5.16(m,1H),4.96(br s,1H),4.92-4.82(m,1H),4.18-3.84(m,4H),3.66(d,J=12.4Hz,1H),3.49(d,J=3.6Hz,1H),3.14-3.04(m,4H),3.00(s,1H),2.96-2.77 (m,2H),2.15-2.08(m,1H),2.02(s,1H),1.99-1.94(m,1H),1.92-1.81(m,2H),1.80-1.68(m,3H),1.56-1.36(m,4H),0.99-0.90(m,3H).

[0393] Example 16: Preparation of Compound 16

[0394]

[0395] Step 1: Synthesis of compound 16

[0396]

[0397] Compound 7-9 (150 mg, 0.74 mmol), compound 14-3 (82.21 mg, 0.22 mmol), palladium acetate (9.08 mg, 0.04 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (46.81 mg, 0.08 mmol), cesium carbonate (263.57 mg, 0.81 mmol) and A mixture of molecular sieves (150 mg) and dioxane (5 mL) was stirred at 110°C under a nitrogen atmosphere for 12 hours. The reaction solution was concentrated to obtain a crude product. Compound 16 was obtained by separation and purification using preparative supercritical fluid chromatography (chromatographic column: SynergiMax-RP 150×30 mm, 4 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; flow rate: 30 mL / min; gradient: 25%-100%, 19 min). MS m / z (ESI): 538.3 [M+H] + .

[0398] 1H NMR(400MHz, CDCl3)δ:7.69(s,1H),7.33(dd,J=12.0,8.0Hz,1H),7.04(s,1H),7.01(d,J=8.0H z,1H),5.49-5.36(m,1H),4.64(d,J=12.0Hz,1H),4.42(d,J=12.0Hz,1H),4.23-4.01(m,3H),3 .94-3.84(m,3H),3.71-3.61(m,1H),3.48-3.40(m,3H),3.20-3.18(m,1H),2.63-2.41(m,4H), 1.95-1.86(m,4H),1.71-1.52(m,3H),0.90-0.87(m,1H),0.65-0.48(m,2H),0.33-0.24(m,2H).

[0399] Example 17: Preparation of Compound 17

[0400]

[0401] Step 1: Synthesis of compound 17

[0402]

[0403] Compound 5-5 (80 mg, 0.33 mmol) was dissolved in dioxane (2 mL), 14-3 (150 mg, 0.40 mmol), palladium acetate (7.5 mg, 0.03 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (39 mg, 0.07 mmol), cesium carbonate (220 mg, 0.67 mmol) were added, and the temperature was raised to 110°C in a nitrogen atmosphere for 2 hours. Water (10 mL) was added for dilution, and the mixture was extracted with ethyl acetate (10 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The mixture was purified by preparative high performance liquid chromatography (chromatographic column: Boston Prime C18, 150×30 mm, 5 μm; mobile phase: 0.225% formic acid aqueous solution-acetonitrile; gradient: acetonitrile 25%-35%, 15 min) to obtain compound 17. MS m / z(ESI):574.1[M+H] + .

[0404] 1HNMR(400MHz,DMSO-d6)δ:7.39(d,J=2.4Hz,1H),7.24(s,1H),7.09(d,J=2.4Hz,1 H),5.27(d,J=54.0Hz,1H),5.09–4.89(m,2H),4.16–3.83(m,4H),3.65–3.60(m,1 H),3.08(s,3H),2.98(d,J=8.8Hz,2H),2.91(d,J=9.2Hz,1H),2.82(d,J=7.6Hz,1 H), 2.13–1.71 (m, 10H), 1.41 (s, 2H), 0.91 (d, J = 6.8Hz, 3H), 0.84 (d, J = 6.8Hz, 3H).

[0405] Biological activity test

[0406] Experimental Example 1: SPR (Surface Plasmon Resonance) Test

[0407] Experimental purpose: To test the binding affinity of the compounds of the present invention to KRAS G12V or KRAS WT by SPR method.

[0408] Experimental method: SPR experiments were performed on the Biacore8k system based on the method described by Tran et al. (PNAS, 2020, 117(7): 3363-3364).

[0409] First, the SA chip (GE Healthcare) was pretreated: 1 mol / L NaCl mixed with 50 mM NaOH was injected three times in a row, each time for 1 minute; then equilibrated with HBST buffer (10 mM HEPES, pH 7.5, 150 mM NaCl, 5 mMMgCl2, 0.5 mM TCEP, 5 μM GDP, 0.05% T20). GDP-bound Biotin-labeled KRAS G12V protein (or other proteins, Beijing Kanglong Chemical, His-KRAS (G12V) -BTN, Uniport: P01116-2 (T2-K169)) was injected. The test compound was diluted with HBST buffer to a maximum concentration of 100 μM, with a final DMSO concentration of 2%, and diluted 1:1 in HBST buffer containing 2% DMSO. Multi-cycle dose response: 30 μL / min flow rate, association time of 90 seconds and dissociation time of 360 seconds for each concentration of analyte; single-cycle dose response: 30 μL / min flow rate, association time of 200 seconds and dissociation time of 2500 seconds. All experiments were performed at 25°C. Sensorgrams were solvent corrected and data fitting was performed using Biacore Insight evaluation software. The experimental results are shown in Table 1.

[0410] Table 1 SPR affinity test results of some compounds of the present invention and KRAS G12V

[0411] Test compound KD / nM Example 6 0.59 Example 10 1.41 Example 11-P2 0.35 Example 12-P1 17.8 Example 12-P2 0.1 Example 13-P1 0.8

[0412] The above results show that the compound of the present invention has good affinity with the target mutant protein.

[0413] Experimental Example 2: Nucleotide exchange assay

[0414] Experimental purpose: To determine the effect of the compounds of the present invention on the function of KRAS G12V protein by detecting the inhibitory effect of the compounds of the present invention on the SOS1-mediated KRAS protein nucleotide exchange process.

[0415] Experimental steps:

[0416] Prepare the detection buffer (25mM HEPES (PH 7.5)), 50mM NaCl aqueous solution, 10mM MgCl2 aqueous solution, 0.01% Brij-35). Use Echo550 to prepare the compound in the new 384 detection plate (Corning, Cat#4512), the highest concentration is 100μM, dilute 3 times, dilute 10 points continuously, repeat twice, and the final DMSO concentration is 1%.

[0417] First, add 5 μL of KRAS G12V (Beijing Kanglong Chemical, His-KRAS (G12V)-BTN, Uniport: P01116-2 (T2-K169)) bound to MANT-GDP (Jenabioscience, Catalog No. NU-205S) to the detection plate, spin at 1000 rpm, and centrifuge for 30 seconds. The plate was placed at room temperature for equilibration for 30 minutes. After 30 minutes, the data was recorded (raw data*).

[0418] Second, add 5 μL of SOS1 (cytoskeleton, Catalog No. CS-GE02-XL) and GppNHp (Abcam, Catalog No. ab146659) mixed reagent to the complex wells and negative control wells of the detection plate, and then add 5 μL of GppNHp reagent to the positive control wells, spin at 1000 rpm, and centrifuge for 30 seconds. Incubate the plate at room temperature for 180 minutes.

[0419] Fluorescence signals were read on a SpectraMax (excitation 360 nm, emission 448 nm) (raw data**).

[0420] Normalized data formula: RFU per well = raw data per well** / raw data per well*. GraphpadPrism was used to perform four-parameter fitting on the data to calculate the IC value of the compound. 50 The results are shown in Table 2.

[0421] Table 2 Nucleotide exchange assay test results of some compounds of the present invention

[0422] Test compound <![CDATA[IC 50 / μM]]> Compound 1 2.4 Compound 4 5.1 Compound 5 1.1 Compound 6 0.04 Compound 8 1.9 Compound 9 3.3 Compound 10 0.05

[0423] The detection showed that the compound of the present invention has a significant inhibitory effect on the functional process of the target mutant protein.

[0424] Experimental Example 3: KRAS G12V-Raf1 interaction assay

[0425] Experimental purpose: To detect the effect of the compounds of the present invention on the KRAS G12V-Raf1 interaction.

[0426] In a 5 mL centrifuge tube, 2.5 times the final concentration of KRAS-G12V or KRAS-WT protein (Beijing Kanglong Chemical, His-KRAS (G12V) -BTN, Uniport: P01116-2 (T2-K169)) and GTP (Sigma, G8877) were mixed and diluted with PPI buffer (Perkin Elmer, 61DB10RDF); the plate was incubated at room temperature for 1 hour;

[0427] The compounds were prepared in new white, opaque-bottomed Optiplate-384-well plates (PerkinElmer, 6007299) using an ultrasonic nanoliter liquid handling system Echo650, with the final concentration of the compounds being up to 10 μM, diluted 3-fold, 10 points serially, repeated twice, and the final DMSO concentration being 0.1%;

[0428] Add 8 μL of KRAS-G12V or KRAS-WT protein and GTP mixture to each well of the corresponding position of the 384-well plate and incubate at room temperature for 0.5 hours. Add 4 μL of GST-Raf1-RBD (BPS bioscience, 100519-1) to each well; mix well and incubate the plate at room temperature for 1 hour. Subsequently, add 4 μL of Anti-GST Tb and 4 μL of Anti 6HIS-XL665 (PerkinElmer; 61GSTTLA, 61HISXLA) to each well, mix well, and incubate at room temperature for 30 minutes. Read the fluorescence signal on the Envision HTS MultilabelReader (excitation 337nm, emission 620nm / 665nm).

[0429] The wells containing 1 μM of the control compound Cpd5 were defined as 100% inhibition (PC, positive control), and the wells containing a final concentration of 0.1% DMSO were defined as 0% inhibition (NC, negative control); inhibition rate % = (Avg.NC-RFU.Cpd) / (Avg.NC-Avg.PC) x 100%. (RFU.Cpd refers to the relative fluorescence value corresponding to each well in the compound group, i.e., emission 620nm / 665nm). Graphpad Prism was used to perform a four-parameter fitting on the data to calculate the IC value of the compound. 50 value.

[0430] The structure of the comparative compound Cpd5 is as follows, which can be prepared with reference to Example 5 in WO2022132200A1:

[0431]

[0432] Table 3 KRAS G12V-Raf1 interaction assay test results of some compounds of the present invention

[0433] Test compound <![CDATA[IC 50 / nM]]> Compound 12-P2 11.5 Compound 13-P1 67

[0434] The detection showed that the compound of the present invention has a significant inhibitory effect on the functional process of the target mutant protein.

[0435] Experimental Example 4: Determination of cell p-ERK inhibitory activity

[0436] Experimental purpose: to detect the inhibitory effect of the compounds of the present invention on cell p-ERK and to evaluate the inhibitory effect of the compounds of the present invention on the KRAS target.

[0437] Experimental steps: Human lung cancer cells NCI-H441 cells (No.: HTB-174, ATCC) were cultured with complete culture medium, i.e. RPMI 1640 medium (Gibco, 11415) containing 10% fetal bovine serum (Corning, 35-081-CV) and 1% Penicillin-Streptomycin solution (Gibco, 15140122).

[0438] NCI-H441 cells were seeded in a new white, bottom-impermeable 384-well plate (Corning, 3570) at a density of 1000 cells / well using complete culture medium, with 40 μL of cell suspension per well; the plates were cultured in a 37° C., 5% CO 2 cell culture incubator overnight.

[0439] Take out the 384-well cell culture plate, add 40nL of the compound to be tested to the cell culture plate using the ultrasonic nanoliter liquid handling system Echo650, so that the final concentration of the compound is up to 1μM, dilute 3 times, dilute 10 points continuously, repeat twice, and the final DMSO concentration is 0.1%. Place in a 37℃, 5% CO2 cell culture incubator for 3 hours.

[0440] Advanced Phospho-ERK 1 / 2 (Thr202 / Tyr204) kit (Cisbio, 64AERPEG) was used to detect the intracellular p-ERK content. Prepare lysis buffer (1x, kit components): 1 volume of lysis buffer (4x, meaning four times the final concentration, i.e., the original concentration was diluted four times) and 3 volumes of ultrapure water for use; prepare premixed antibody solution: 1 volume of Eu Cryptate antibody or d2 antibody and 19 volumes of detection buffer for use.

[0441] Remove the 384-well plate and discard the supernatant; add 16μL lysis buffer (1x), incubate at room temperature with shaking for at least 30 minutes; centrifuge at 3000rpm for 2 minutes. Add 4μL Premixed antibody solution to each well and mix for 30 seconds; centrifuge at 1000rpm for 5 minutes. Incubate at room temperature for 2.5 hours. Read the fluorescence signal on the Envision HTS Multilabel Reader (excitation 337nm, emission 620nm / 665nm).

[0442] The wells containing 1 μM Staurosporine (Test Yang Shen, MCE, Catalog No. HY-15141) were defined as 100% inhibition (PC, positive control), and the wells containing a final concentration of 0.1% DMSO were defined as 0% inhibition (NC, negative control); inhibition rate % = (Avg.NC-RFU.Cpd) / (Avg.NC-Avg.PC) x 100% (RFU.Cpd refers to the relative fluorescence value corresponding to each well in the compound group, i.e., emission 620nm / 665nm). The data were analyzed by concentration-inhibition rate nonlinear regression analysis in GraphPad Prism 9 to obtain the IC values ​​of the test compounds. 50 Values, data see Table 4.

[0443] The detection showed that the compound of the present invention has a significant inhibitory effect on the p-ERK function of the target cells.

[0444] Experimental Example 5: Determination of cell antiproliferative activity

[0445] Experimental purpose: To detect the inhibitory effect of the compounds of the present invention on the proliferation of human lung cancer cells NCI-H441.

[0446] Experimental steps: Human lung cancer cells NCI-H441 (No. HTB-174, ATCC) were cultured with complete culture medium, RPMI 1640 medium (Gibco, 22400089) containing 10% fetal bovine serum (Corning, 35-081-CV) and 1% Penicillin-Streptomycin solution (Gibco, 15140122).

[0447] The cells were seeded in a new 384-well cell culture plate (Greiner, 781091) at 1000 cells per well using complete culture medium, with 60 μL of cell suspension per well; cultured overnight in a 37°C, 5% CO2 cell culture incubator. The compound to be tested was diluted with complete culture medium, 20 μL was added to each well, so that the final concentration of the compound was up to 10 μM, diluted 3 times, and diluted 10 points in a row.

[0448] Repeat twice, the final DMSO concentration is 0.25%. Centrifuge at 1000 rpm for 1 min at room temperature. Place in a 37°C, 5% CO2 cell culture incubator for 5 days.

[0449] The 384-well cell culture plate was removed and 40 μL of supernatant was discarded. Then, 20 μL of luminescent cell activity detection reagent (Promega, G7573) was added, and the plate was shaken in the dark for 2 minutes, and then centrifuged at 1000 rpm for 1 minute. The plate was placed in the dark at room temperature for 10 minutes. The luminescent signal value (Lumi.Cpd) was read on the Envision HTS Multilabel Reader.

[0450] The wells containing 1 μM Staurosporine (Test Yang Shen, MCE, Catalog No. HY-15141) were defined as 100% inhibition (PC, positive control), and the wells containing 0.25% DMSO at a final concentration were defined as 0% inhibition (NC, negative control); inhibition rate % = (Avg.NC-Lumi.Cpd) / (Avg.NC-Avg.PC) x 100%. The data were analyzed by concentration-inhibition rate nonlinear regression analysis in GraphPad Prism 9 to obtain the IC value of the compound. 50 The specific experimental data are shown in Table 4.

[0451] The test results show that the compound of the present invention has a significant inhibitory effect on the proliferation process of target cells.

[0452] Table 4 IC values ​​of some compounds of the present invention for NCI H441 p-ERK / CTG 50 Test Results

[0453] Test compound p-ERK / nM CTG / nM Compound 13-P2(0093-P2) 45 63 Compound 14-P1(0096-P1) 72 83

Claims

1. A compound of formula I or a pharmaceutically acceptable salt or stereoisomer thereof; in, X1, X2, X3 are selected from N, C-Ra; Ra is selected from H, halogen atoms, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 alkoxy; L1 is selected from O, S, NH; Ring B is a 5-6 membered aryl group or a 5-6 membered heteroaryl group, and Ring B may be optionally substituted by one or more groups selected from the following groups: F, Cl, Br, CN, OH, NH2, C 1-3 Alkyl, C 1-3 alkyl halide; R1 is a 6-12 membered nitrogen-containing hetero-bridged ring substituted C 1-3 Alkyl; the nitrogen-containing hetero bridged ring is a bicyclic bridged ring, and may be optionally substituted by one or more groups selected from the following: F, Cl, CN, NH2, OH, C 1-3 Alkyl, C 1-3 alkyl halide; R2 is selected from H, CN, halogen atoms, C 1-3 Alkyl, C 1-3 Haloalkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl, hydroxy-C 1-3 alkyl; R3 is selected from H, CN, halogen atoms, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, hydroxy-C 1-3 alkyl; R x R5 is selected from C 1-3 alkyl, 3-7 membered cycloalkyl substituted by R5, 3-7 membered heterocycloalkyl substituted by R5, 5-6 membered aryl substituted by R5, 5-6 membered heteroaryl substituted by R5; R5 is selected from H, CN, OH, halogen atom, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 3-6 Cycloalkyl, C 3-6 heterocycloalkyl; Or R3, Rx and the carbon atom to which they are connected are cyclized together to form a 5-6 membered aryl or 5-6 membered heteroaryl; the 5-6 membered aryl or the 5-6 membered heteroaryl may be optionally replaced by R6 and R 6a Substituted C 1-3 alkyl; R6 is selected from H, 5-6 membered aryl, 5-6 membered heteroaryl; and R6 may be optionally substituted by one or more of the following groups: H, NH2, CN, halogen atoms, C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, hydroxy-C 1-3 Alkyl; the R 6a Selected from C 1-3 Alkyl, C 1-3 Halogenated alkyl, C 1-3 Alkoxy, hydroxy-C 1-3 alkyl.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: Ring B is substituted by OH; or ring B is substituted by Cl; or ring B is substituted by both OH and Cl.

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: At least one of X1 and X2 is N; or both X1 and X2 are N.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: X3 is N or C-Ra; preferably X3 is N; Ra has the same meaning as in the above formula I, preferably H, C 1-3 alkyl.

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: X1, X2, and X3 are all N.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: L1 is O, or L1 is S, or L1 is NH.

7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: Ring B is a benzene ring, and may be optionally substituted by Cl or / and OH.

8. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: Has the following structure:

9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: R1 is the following group substituted by one or more halogen atoms; Preferably, R1 is selected from the following groups:

10. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: R2 is selected from methyl, ethyl, isopropyl, and cyclopropyl.

11. The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: R3 is H.

12. The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The specific structure is as follows: Among them, R6, R 6a has the same meaning as in the above formula I; X4 is N or C-Ra, Ra has the same meaning as in the above formula I; preferably, X4 is N or CH; R 6a Selected from H, methyl, trifluoromethyl; R 6a Selected from pyridyl and pyrazolyl; in one technical solution, R6 is selected from 1-methylpyrazol-2-yl and 2-aminopyridin-3-yl.

13. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The 5-6 membered aryl group in R6 is phenyl; preferably, the 5-6 membered heteroaryl group in R6 is selected from pyridine and pyrazolyl; in any of the above technical solutions of formula I of the present invention, R6 is selected from 2-amino-pyridin-3-yl and 1-methylpyrazol-3-yl.

14. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: The compound has the following structure: Among them, X1, X2, X3, L1, R1, R2, R6, R 6a has the same meaning as in formula I in claim 1.

15. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that: In the present invention, the compound represented by formula I is selected from the following compounds or pharmaceutically acceptable salts thereof: 16 . A pharmaceutical composition comprising the compound according to claim 1 , or a pharmaceutically acceptable salt thereof, a stereoisomer thereof, and any pharmaceutically acceptable carrier.

17. Use of the compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt or stereoisomer thereof in the preparation of a medicament for treating diseases associated with KRAS G12V gene mutation.

18. Use of the pharmaceutical composition according to claim 16 in the preparation of a medicament for treating diseases associated with KRAS G12V gene mutation.

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