WO2025067316A1 - Benzylamino-substituted tricyclic heterocyclic compound, composition thereof, preparation thereof and use thereof - Google Patents

Abstract

The present invention belongs to the technical field of pharmaceutical chemistry, and relates to a benzylamino-substituted tricyclic heterocyclic compound as represented by formula I, a composition thereof, a preparation thereof and the use thereof. Specifically, the general formula of the tricyclic heterocyclic ring has a structure as represented by formula I. The compound of the present invention has excellent in-vitro inhibitory activity against SOS1, and has good in-vivo inhibitory activity against an NCI-H358 subcutaneous xenograft tumor. In addition, the compound of the present invention has characteristics such as relatively low plasma clearance, relatively long drug half-life, relatively good exposure and relatively high bioavailability, has good pharmacokinetic properties, does not show strong inhibition against various CYP enzyme subtypes, is high in terms of levels of safety, has in-vivo anti-tumor activity, and is relatively low in terms of levels of toxicity.

Description

Benzylamino-substituted tricyclic heterocyclic compounds and compositions, preparations and uses thereof Technical Field

The invention belongs to the technical field of pharmaceutical chemistry, and relates to a series of novel benzylamino-substituted tricyclic heterocyclic compounds, pharmaceutical compositions and pharmaceutical preparations containing the same, and medical uses thereof.

Background Art

KRAS gene mutations are common in pancreatic cancer, lung adenocarcinoma, colorectal cancer, gallbladder cancer, bile duct cancer and thyroid cancer. It is a GTP-binding protein. RAS has two main forms in the body: an inactive state bound to GDP and an activated state bound to GTP. Its activity is regulated by two proteins. Guanylate exchange factors (GEFs) such as SOS1 promote the release of GDP from RAS proteins, allowing GTP to bind and activate RAS; GTPase activating proteins activate the GTPase activity of RAS proteins, hydrolyzing GTP bound to RAS proteins into GDP, inactivating RAS. When in the GTP-bound state, RAS family proteins are active and engage effector proteins (including RAF and PI3K) to promote RAF/MEK/ERK, PI3K/AKT/mTOR and other pathways. These pathways affect a variety of cellular processes, such as proliferation, survival, metabolism, etc.

SOS1 has two binding sites for RAS family proteins: one is a catalytic site that binds to GDP-bound RAS family proteins to promote guanine nucleotide exchange, and the other is an allotropic site that binds to GTP-bound RAS family proteins to further increase the catalytic GEF function of SOS1 (Biochem. Pharmacol., 2011, 82(9): 1049-1056). SOS1 is important in the activation of mutant KRAS and oncogenic signaling in cancer (Nat. Commun., 2012, 3: 1168). In tumor cells carrying KRAS mutations, reducing SOS1 content can reduce the proliferation rate of tumor cells, while no effect is observed in KRAS wild-type cell lines.

As the first confirmed oncogene, RAS is the oncogene with the highest mutation rate, accounting for 25% of human cancers. In recent decades, the interaction between RAS family proteins and SOS1 proteins has been increasingly recognized. Currently, only Boehringer Ingelheim's SOS1 inhibitor BI1701963 has entered Phase I clinical trials, and no SOS1 inhibitors have been developed and marketed. Therefore, the development of new SOS1 inhibitors has great clinical value and broad market prospects.

Summary of the invention

Problem that the invention aims to solve

In order to develop new SOS1 inhibitors, the present invention aims to provide a novel benzylamino-substituted tricyclic heterocyclic compound and its composition, preparation and use, wherein the derivative has excellent SOS1 inhibitory activity in vitro, good in vivo pharmacodynamics and pharmacokinetic properties, and high safety.

Solutions for solving problems

In a first aspect, the present invention provides a compound having a structure of Formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof:

in:

Every independently represent a single bond or a double bond;

X is selected from CR ₆ , C(R ₆ ) ₂ and C(═O);

Y is selected from CR ₇ and C(R ₇ ) ₂ ;

z is selected from N and NR ⁸ , or Z is absent; when Z is absent, Y is directly connected to the phthalazine ring via a single bond;

Ring A is selected from phenyl and thienyl;

each R ¹ is independently selected from methyl, ethyl, phenyl, halogen and cyano, wherein the methyl, ethyl or phenyl is optionally substituted with one or more substituents selected from methylaminomethyl, dimethylaminomethyl, hydroxyl and halogen;

^R2 and ^R3 are independently hydrogen;

^R4 is methyl;

R ⁵ is selected from C _1-8 alkyl, C _3-6 cycloalkyl and 3-8 membered heterocyclyl, wherein the C _1-8 alkyl is optionally substituted by one or more deuterium; the 3-8 membered heterocyclyl contains a heteroatom selected from O, N and S, and the 3-8 membered heterocyclyl is optionally substituted by one or more C _1-8 alkyl;

If present, each R ⁶ is independently hydrogen;

Each R ⁷ is independently selected from hydrogen and methyl; or, two R ⁷ together with the carbon atom to which they are attached form a cyclopropyl group;

If present, R ⁸ is selected from methyl and cyclopropyl, wherein the methyl is optionally substituted with one or more deuterium;

n is 1 or 2;

Furthermore, when Z is absent, Y is C(R ₇ ) ₂ , and X is C(═O), the two R ⁷ are not connected to each other and are different from each other;

Also, does not contain the following compounds:

In a second aspect, the present invention provides the following non-limiting examples of the above-mentioned compounds having the structure of formula I:

(1) 2-methyl-3-((1R)-1-((1,2,4,9-tetramethyl-3-oxo-1,2,3,4-tetrahydropyridazin[4,5-g]quinoxalin-6-yl)amino)ethyl)benzonitrile;

(2) 9-(((R)-1-(3-(1,1-difluoro-2-hydroxyethyl)-2-fluorophenyl)ethyl)amino)-1,3,4,6-tetramethyl-3,4-dihydropyridazin[4,5-g]quinoxalin-2(1H)-one;

(3) 9-(((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-3,6-dimethyl-1,4-bis(methyl-d ₃ )-3,4-dihydropyridazino[4,5-g]quinoxalin-2(1H)-one;

(4) (R)-2-methyl-3-(1-((1,2,2,4,9-pentamethyl-3-oxo-1,2,3,4-tetrahydropyridazin[4,5-g]quinoxalin-6-yl)amino)ethyl)benzonitrile;

(5) (R)-9-((1-(3-(1,1-difluoro-2-hydroxyethyl)-2-fluorophenyl)ethyl)amino)-1,3,3,4,6-pentamethyl-3,4-dihydropyridazin[4,5-g]quinoxalin-2(1H)-one;

(6) (R)-1,3,3,4,6-pentamethyl-9-((1-(5-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-3,4-dihydropyridazin[4,5-g]quinoxalin-2(1H)-one;

(7) (R)-9-((1-(5-(2-((dimethylamino)methyl)phenyl)thiophen-2-yl)ethyl)amino)-1,3,3,4,6-pentamethyl-3,4-dihydropyridazin[4,5-g]quinoxalin-2(1H)-one;

(8) 3-((1R)-1-((4-cyclopropyl-1,2,9-trimethyl-3-oxo-1,2,3,4-tetrahydropyridazin[4,5-g]quinoxalin-6-yl)amino)ethyl)-2-methylbenzonitrile;

(9) 3-((1R)-1-((1-cyclopropyl-2,4,9-trimethyl-3-oxo-1,2,3,4-tetrahydropyridazin[4,5-g]quinoxalin-6-yl)amino)ethyl)-2-methylbenzonitrile;

(10) (R)-2-methyl-3-(1-((1′,4′,9′-trimethyl-3′-oxo-3′,4′-dihydro-1′H-spiro[cyclopropane-1,2′-pyridazinone[4,5-g]quinoxaline]-6′-yl)amino)ethyl)benzonitrile;

(11) (R)-2-methyl-3-(1-((2,4,9-trimethyl-3-oxo-3,4-dihydropyridazin[4,5-g]quinoxalin-6-yl)amino)ethyl)benzonitrile;

(12) N-((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-5-methyl-1-(tetrahydrofuran-3-yl)-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-amine;

(13) 2-methyl-3-((1R)-1-((5-methyl-1-(tetrahydrofuran-3-yl)-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)benzonitrile;

(14) 2-methyl-3-((1R)-1-((5-methyl-1-(tetrahydrofuran-3-yl)-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)benzonitrile;

(15) N-((R)-1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-5-methyl-1-(tetrahydrofuran-3-yl)-1H-pyrrolo[2,3-g]phthalazin-8-amine;

(16) 2-methyl-3-((1R)-1-((3,3,5-trimethyl-1-(tetrahydrofuran-3-yl)-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)benzonitrile;

(17) (R)-3-(1-((1-isopropyl-5-methyl-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)-2-methylbenzonitrile;

(18) (R)-3-(1-((1-cyclobutyl-5-methyl-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)-2-methylbenzonitrile;

(19) (R)-3-(1-((1-cyclopentyl-5-methyl-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)-2-methylbenzonitrile;

(20) 2-methyl-3-((1R)-1-((5-methyl-1-(tetrahydro-2H-pyran-3-yl)-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)benzonitrile;

(21) (R)-2-methyl-3-(1-((5-methyl-1-(tetrahydro-2H-pyran-4-yl)-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)benzonitrile;

(22) 2-methyl-3-((1R)-1-((5-methyl-1-(1-methylpyrrolidin-3-yl)-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-yl)amino)ethyl)benzonitrile; and

(23) 5-Methyl-N-((R)-1-(4-(2-((methylamino)methyl)phenyl)thiophen-2-yl)ethyl)-1-(tetrahydrofuran-3-yl)-2,3-dihydro-1H-pyrrolo[2,3-g]phthalazin-8-amine.

In a third aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned compound having the structure of Formula I or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, and at least one pharmaceutically acceptable carrier.

Preferably, in the above-mentioned pharmaceutical composition, the pharmaceutically acceptable carrier includes (but is not limited to) diluents (or fillers), binders, disintegrants, lubricants, wetting agents, thickeners, glidants, flavoring agents, olfactory agents, preservatives, antioxidants, pH adjusters, solvents, cosolvents, surfactants, opacifiers (opacifiers), etc.

In a fourth aspect, the present invention provides a pharmaceutical preparation, which is made from the above-mentioned compound having the structure of Formula I or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, or is made from the above-mentioned pharmaceutical composition.

In a fifth aspect, the present invention provides the use of the above-mentioned compound having the structure of Formula I or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation in the preparation of a drug for preventing and/or treating a disease mediated at least in part by the SOS1 protein.

Preferably, in the above-mentioned use, the disease at least partially mediated by the SOS1 protein is cancer, in particular a cancer selected from pancreatic cancer, lung cancer, colorectal cancer, bile duct cancer, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial carcinoma, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcoma.

In a sixth aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned compound having a structure of Formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation, and at least one additional cancer therapeutic agent.

In one or more embodiments, the cancer therapeutic agent is selected from trametinib and cediranib.

In the seventh aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned compound having the structure of Formula I or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation, and at least one additional cancer therapeutic agent.

In an eighth aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation, wherein the compound is used As medicine.

In a ninth aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation, which is used to prevent and/or treat a disease mediated at least in part by SOS1 protein.

In the tenth aspect, the present invention provides a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation, which is used for preventing and/or treating cancer.

In an eleventh aspect, the present invention provides a method for preventing and/or treating a disease mediated at least in part by SOS1 protein, comprising administering a preventive and/or therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation to an individual in need thereof.

In a twelfth aspect, the present invention provides a method for preventing and/or treating cancer, which comprises administering a preventive and/or therapeutically effective amount of a compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or the above-mentioned pharmaceutical composition, or the above-mentioned pharmaceutical preparation to an individual in need thereof.

In one or more embodiments, the disease mediated at least in part by the SOS1 protein is cancer, in particular a cancer selected from pancreatic cancer, lung cancer, colorectal cancer, bile duct cancer, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial carcinoma, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcoma.

Effects of the Invention

The compounds of the present invention have excellent in vitro inhibitory activity against SOS1 and good in vivo inhibitory activity against NCI-H358 subcutaneous xenograft tumors; in addition, the compounds of the present invention have the characteristics of low plasma clearance, long drug half-life, good exposure, high bioavailability, etc., have good pharmacokinetic properties, and do not show strong inhibition on various CYP enzyme subtypes, and have high safety. In addition, the compounds of the present invention and the compositions thereof with other anticancer agents have good in vivo tumor inhibitory activity. In addition, the compounds of the present invention have low toxicity to various organs.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 is a TGI graph of tumor growth inhibition rates of compound 1, compound 13, compound 14 and positive control compound 1.

FIG2 is a graph showing the inhibition of tumor volume changes by compound 1, compound 13, compound 14 and positive control compound 1.

DETAILED DESCRIPTION

Before the present invention is further described, it is to be understood that the present invention is not limited to the particular embodiments described herein; it is also to be understood that the terminology used herein is for the purpose of describing only and not limiting the particular embodiments.

[Definition of terms]

Unless otherwise stated, the following terms have the following meanings.

The term "pharmaceutically acceptable salt" refers to a salt of a compound having the structure of Formula I that is substantially non-toxic to an organism. Pharmaceutically acceptable salts generally include (but are not limited to) salts formed by reacting a compound of the present invention with a pharmaceutically acceptable inorganic acid or organic acid, and such salts are also known as acid addition salts. Common inorganic acids include (but are not limited to) hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, sulfuric acid (which can form sulfates or acid sulfates), phosphoric acid (which can form phosphates or acid phosphates), etc. Common organic acids include (but are not limited to) trifluoroacetic acid, citric acid (which can form citric acid monosalt, disalt or trisalt), maleic acid (which can form maleic acid monosalt or disalt), fumaric acid (which can form fumaric acid monosalt or disalt), succinic acid (which can form succinic acid monosalt or disalt), tartaric acid (which can form tartaric acid monosalt or disalt), oxalic acid (which can form oxalic acid monosalt or disalt), malonic acid (which can form malonic acid monosalt or disalt), malic acid (which can form malic acid monosalt or disalt), oxalic acid (which can form oxalic acid monosalt or disalt), lactic acid, pyruvic acid, salicylic acid, formic acid, acetic acid, propionic acid, benzoic acid, glycolic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.

The term "hydrate" refers to a compound of the present invention or a pharmaceutically acceptable salt thereof bound by water through non-covalent intermolecular forces. Common hydrates include (but are not limited to) hemihydrate, monohydrate, dihydrate, trihydrate, etc.

The term "solvate" refers to a substance formed by the combination of a compound of the present invention or a pharmaceutically acceptable salt thereof and at least one solvent molecule through non-covalent intermolecular forces. The term "solvate" includes "hydrate". Common solvates include (but are not limited to) hydrates, ethanolates, acetonates, etc. It should be understood that the present invention encompasses all solvate forms having SOS1 inhibitory activity.

The term "isomers" refers to compounds that have the same number and types of atoms and therefore the same molecular weight, but differ in the arrangement or configuration of the atoms in space.

The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has a vertical asymmetric plane due to at least one chiral factor (including chiral center, chiral axis, chiral plane, etc.), thereby being able to rotate plane polarized light. Since there are asymmetric centers and other chemical structures that may cause stereoisomerism in the compounds of the present invention, the present invention also includes these stereoisomers and mixtures thereof. Since the compounds of the present invention and their salts include asymmetric carbon atoms, they can exist in the form of a single stereoisomer, a racemate, an enantiomer, and a mixture of diastereomers. Generally, these compounds can be prepared in the form of a racemic mixture. However, if desired, such compounds can be prepared or separated to obtain pure stereoisomers, i.e., single enantiomers or diastereomers, or single stereoisomer-enriched (purity ≥98%, ≥95%, ≥93%, ≥90%, ≥88%, ≥85% or ≥80%) mixtures. As described below, a single stereoisomer of a compound is prepared synthetically from an optically active starting material containing the desired chiral center, or by preparing a mixture of enantiomeric products followed by separation or resolution, such as conversion to a mixture of diastereoisomers followed by separation or recrystallization, chromatography, use of a chiral resolution agent, or direct separation of the enantiomers on a chiral chromatographic column. Starting compounds with a specific stereochemistry are either commercially available or prepared as described below and resolved by methods well known in the art. The term "enantiomer" refers to a pair of stereoisomers that are non-superimposable mirror images of each other. The term "diastereomer" or "diastereomers" refers to optical isomers that are not mirror images of each other. The term "racemic mixture" or "racemate" refers to a mixture containing equal parts of a single enantiomer (i.e., an equimolar mixture of two R and S enantiomers). The term "non-racemic mixture" refers to a mixture containing unequal parts of a single enantiomer. Unless otherwise stated, all stereoisomeric forms of the compounds of the present invention are within the scope of the present invention.

The term "tautomer" (or "tautomeric form") refers to structural isomers with different energies that can be interconverted through a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (or prototropic tautomers) include (but are not limited to) interconversions through proton migration, such as keto-enol isomerization, imine-enamine isomerization, amide-imino alcohol isomerization, etc. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

The term "cis-trans isomer" refers to stereoisomers formed by the different positions of atoms (or groups) located on both sides of a double bond or ring system relative to a reference plane; in cis isomers, the atoms (or groups) are located on the same side of the double bond or ring system, and in trans isomers, the atoms (or groups) are located on opposite sides of the double bond or ring system. Unless otherwise indicated, all cis-trans isomeric forms of the compounds of the present invention are within the scope of the present invention.

The term "isotope-labeled substance" refers to a compound formed by replacing a specific atom in a structure with an isotope atom thereof. Unless otherwise indicated, the compounds of the present invention include various isotopes of H, C, N, O, F, P, S, and Cl, such as ² H(D), ³ H(T), ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, ¹⁸ F, ³¹ P, ³² P, ³⁵ S, ³⁶ S, and ³⁷ Cl.

The term "prodrug" refers to a derivative compound that can directly or indirectly provide a compound of the present invention after being applied to a patient. Particularly preferred derivative compounds or prodrugs are compounds that can increase the bioavailability of the compound of the present invention when administered to a patient (e.g., more easily absorbed into the blood), or compounds that promote the delivery of the parent compound to the site of action (e.g., the lymphatic system). Unless otherwise indicated, all prodrug forms of the compounds of the present invention are within the scope of the present invention, and various prodrug forms are well known in the art.

The term "alkyl" refers to a straight or branched chain saturated aliphatic hydrocarbon group of 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8) carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl and various branched chain isomers thereof.

The term "cycloalkyl" refers to a cyclic saturated aliphatic hydrocarbon group of 3 to 6 (e.g., 3, 4, 5, 6) carbon atoms, which may be monocyclic or bicyclic. Non-limiting examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term "heterocyclic group" refers to a 3- to 8-membered (e.g., 3, 4, 5, 6, 7, 8-membered) cyclic saturated and/or unsaturated aliphatic hydrocarbon group, which may be It is a monocyclic or bicyclic ring, which may contain one or more (eg, 1, 2, 3 or 4) heteroatoms selected from N, O or S, and may be a monocyclic, fused, bridged or spirocyclic ring.

The term "methyl" refers to a _-CH3 or -Me group.

The term "ethyl" refers to _{a -CH2CH3} , _-C2H5 or _-Et group.

The term "isopropyl" refers to a -CH(CH ₃ ) ₂ or -i-Pr group.

The term "cyclopropyl" refers to Or a -cyc-Pr group.

The term "cyclobutyl" refers to Or a -cyc-Bu group.

The term "cyclopentyl" refers to Or a -cyc-amyl group.

The term "tetrahydropyrrolyl" or "pyrrolidine" refers to Group; Accordingly, the term "tetrahydropyrrol-3-yl" or "pyrrolidin-3-yl" refers to Group.

The term "tetrahydrofuranyl" refers to group; accordingly, the term "tetrahydrofuran-3-yl" refers to Group.

The term "tetrahydropyranyl" refers to Group; accordingly, the term "tetrahydropyran-3-yl" refers to The term "tetrahydropyran-4-yl" refers to Group.

The term "phenyl" refers to -C ₆ H ₅ or -Ph group.

The term "thienyl" refers to or -C ₄ H ₃ S group; accordingly, the term "thiophen-2-yl" refers to Group.

The term "phthalazine" refers to Accordingly, the term "phthalazine ring" refers to a ring system derived from phthalazine.

The term "halogen" refers to fluorine (F), chlorine (Cl), bromine ( _Br ) and iodine (I) located in main group VII of the periodic table.

The term "hydroxy" refers to an -OH group.

The term "cyano" refers to a -CN group.

The term "methylaminomethyl" refers to a _{-CH2NHCH3 group} ; correspondingly, the term "dimethylaminomethyl" refers to a _-CH2N ( _CH3 ) ₂ group.

The term "single bond" refers to a covalent chemical bond between ^sp3 hybridized atoms for connecting or interacting with each other; correspondingly, the term "double bond" refers to a covalent chemical bond between ^sp2 hybridized atoms.

The term "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes the occurrence of said event or circumstance and The event or situation does not occur. For example, R _4d is "optionally" substituted with halogen, which means that R _4d may be unsubstituted, or may be monosubstituted, polysubstituted or fully substituted with halogen atoms. It will be understood by those skilled in the art that for any group containing one or more substituents, no substitution or substitution pattern that is sterically impossible to exist and/or cannot be synthesized will be introduced.

[Benzylamino-substituted tricyclic heterocyclic compounds]

The present invention provides a series of novel benzylamino-substituted tricyclic heterocyclic compounds or pharmaceutically acceptable forms thereof, such as salts, hydrates, solvates, stereoisomers, tautomers, cis-trans isomers, isotope-labeled substances or prodrugs of the compounds.

In one embodiment of the present invention, the structure of the compound is as shown in Formula I:

in:

Every independently represent a single bond or a double bond;

X is selected from CR ₆ , C(R ₆ ) ₂ and C(═O);

Y is selected from CR ₇ and C(R7) ₂ ;

Z is selected from N and NR ⁸ , or Z is absent; when Z is absent, Y is directly connected to the phthalazine ring via a single bond;

Ring A is selected from phenyl and thienyl (especially thien-2-yl);

each R ¹ is independently selected from methyl, ethyl, phenyl, halogen and cyano, wherein the methyl, ethyl or phenyl is optionally substituted with one or more substituents selected from methylaminomethyl, dimethylaminomethyl, hydroxyl and halogen;

^R2 and ^R3 are independently hydrogen;

^R4 is methyl;

R ⁵ is selected from C _1-8 alkyl, C _3-6 cycloalkyl and 3-8 membered heterocyclyl, wherein the C _1-8 alkyl is optionally substituted by one or more deuterium; the 3-8 membered heterocyclyl contains a heteroatom selected from O, N and S, and the 3-8 membered heterocyclyl is optionally substituted by one or more C _1-8 alkyl;

If present, each R ⁶ is independently hydrogen;

Each R ⁷ is independently selected from hydrogen and methyl; or, two R ⁷ together with the carbon atom to which they are attached form a cyclopropyl group;

If present, R ⁸ is selected from methyl and cyclopropyl, wherein the methyl is optionally substituted with one or more deuterium;

n is 1 or 2;

Furthermore, when Z is absent, Y is C(R ₇ ) ₂ , and X is C(═O), the two R ⁷ are not connected to each other and are different from each other;

Also, does not contain the following compounds:

"If present, each R ⁶ is independently hydrogen" means that when X is selected from CR ₆ or C(R ₆ ) ₂ , each R ⁶ is independently hydrogen.

In a preferred embodiment of the present invention, ring A in formula I is selected from phenyl and thiophen-2-yl.

In a preferred embodiment of the present invention, each R ¹ in Formula I is independently selected from -CH ₃ , -CH ₂ F, -CHF 2 , -CF 3 , -CH _{2 OH, -CH 2 CH 3 , -CHFCH 3 , -CF 2 CH 3} , -CH 2 CH _{2 OH} , -CHFCH 2 OH, -CF ₂ CH 2 _OH , -F, -Cl, -Br, -Br, -Br _{, -Br} , _-Br , -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br _, -Br, _-Br , -Br, _-Br , -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br, -Br, _-

In a more preferred embodiment of the present invention, each R ¹ in Formula I is independently selected from -CH ₃ , -CHF ₂ , -CF ₂ CH ₂ OH, -F,

In a preferred embodiment of the present invention, ring A in formula I is thiophene-2-yl, n is 1, and R ¹ is substituted at any substitutable position of A, preferably as shown in formula IA or formula IA':

in:

Every independently represent a single bond or a double bond;

R ¹ is selected from

X, Y, Z and R ⁵ are as defined in Formula I.

In another preferred embodiment of the present invention, ring A in formula I is phenyl, n is 2, and two R ^1s are independently substituted at any substitutable position of A, preferably as shown in formula IB or formula IB':

in:

Every independently represent a single bond or a double bond;

each R ¹ is independently selected from -CH ₃ , -CHF ₂ , -CF ₂ CH ₂ OH, -F and -CN;

X, Y, Z and R ⁵ are as defined in Formula I.

In a more preferred embodiment of the present invention, Selected from Best

In a preferred embodiment of the present invention, in Formula I, Formula IA, Formula IA', Formula IB or Formula IB' Selected from

In a more preferred embodiment of the present invention, in Formula I, Formula IA, Formula IA', Formula IB or Formula IB' Selected from

In a preferred embodiment of the present invention, ^R5 in formula I is selected from _C1-4 alkyl, _C3-6 cycloalkyl and 3-6 membered heterocyclyl, wherein the _C1-4 alkyl is optionally substituted by one or more deuterium; the 3-6 membered heterocyclyl contains a heteroatom selected from O and N, and the 3-6 membered heterocyclyl is optionally substituted by one or more _C1-4 alkyl.

In a preferred embodiment of the present invention, R ⁵ in Formula I is selected from -CH ₃ , -CH ₂ D, -CHD ₂ , -CD ₃ , isopropyl, cyclopropyl 4-yl, cyclobutyl, cyclopentyl, cyclohexyl, pyrrolidin-3-yl, 1-methylpyrrolidin-3-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl and tetrahydrofuran-3-yl.

In a more preferred embodiment of the present invention, ^R5 in Formula I is selected from _-CH3 , _-CD3 , isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, 1-methylpyrrolidin-3-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl and tetrahydrofuran-3-yl.

In a preferred embodiment of the present invention, R ⁸ in formula I is selected from -CH ₃ , -CH ₂ D, -CHD ₂ , -CD ₃ and cyclopropyl.

In a more preferred embodiment of the present invention, R ⁸ in Formula I is selected from -CH ₃ , -CD ₃ and cyclopropyl.

In a preferred embodiment of the present invention, in Formula IA or Formula IA' for The compound at this time is as shown in Formula IA-1 or Formula IA'-1:

wherein: R ¹ is as defined in Formula IA or Formula IA'.

In a preferred embodiment of the present invention, in Formula IA or Formula IA' for The compound at this time is shown in Formula IA-2 or Formula IA'-2:

wherein: R ¹ is as defined in Formula IA or Formula IA'.

In a preferred embodiment of the present invention, in Formula IB or Formula IB' Selected from In this case, the compound is represented by any one of Formulas IB-1 to IB-16 or any one of Formulas IB'-1 to IB'-16:

wherein: R ¹ is as defined in Formula IB or Formula IB'.

In a more preferred embodiment of the present invention, in Formula IB or Formula IB' Selected from In this case, the compound is represented by any one of Formula IB-1, Formula IB-2 and Formula IB-5 or any one of Formula IB'-1, Formula IB'-2 and Formula IB'-5, wherein: R ¹ is as defined in Formula IB or Formula IB'.

Specifically, the benzylamino-substituted tricyclic heterocyclic compounds of the present invention include (but are not limited to) the following compounds:

[Drug composition]

The term "pharmaceutical composition" refers to a composition that can be used as a medicament, which comprises a pharmaceutically active ingredient (API) and optionally one or more pharmaceutically acceptable carriers.

The term "pharmaceutically acceptable carrier" refers to a pharmaceutical excipient that is compatible with the active ingredient of the drug and is harmless to the subject, including (but not limited to) one or more of a diluent (or filler), a binder, a disintegrant, a lubricant, a wetting agent, a thickener, a glidant, a flavoring agent, an olfactory agent, a preservative, an antioxidant, a pH adjuster, a solvent, a cosolvent, a surfactant, a light-shielding agent (opacifying agent), etc.

The present invention provides a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof.

In one embodiment of the present invention, the above pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier.

The term "pharmaceutical composition" may also include a compound of the invention or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotopically labeled or prodrug thereof, or a pharmaceutical composition of the invention or a pharmaceutical formulation of the invention, and at least one additional cancer therapeutic agent.

The term "cancer therapeutic agent" refers to a pharmaceutical composition or pharmaceutical formulation that is effective in controlling and/or combating cancer.

In one embodiment of the present invention, the compound of the present invention or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope label or prodrug, or the pharmaceutical composition of the present invention or the pharmaceutical preparation of the present invention can be administered alone or in combination with other cancer therapeutic agents (or anti-tumor drugs). The combined therapy can be achieved by administering different cancer therapeutic agents simultaneously, sequentially or separately.

In one embodiment of the present invention, the cancer is pancreatic cancer, non-small cell lung cancer, or colorectal cancer.

[Drug preparations]

The term "pharmaceutical preparation" refers to a finished drug prepared in a certain form for use by patients.

The present invention provides a pharmaceutical preparation, which is prepared from the pharmaceutical composition of the present invention.

In one embodiment of the present invention, the above-mentioned pharmaceutical preparation is a solid preparation for oral administration, including (but not limited to) pharmaceutically acceptable capsules, tablets, pills, powders, granules, etc. The solid preparation can be coated or microencapsulated with a coating or shell material (such as an enteric coating or other materials known in the art). The solid preparation can contain an opacifying agent, and the active ingredient therein can be released in a certain part of the digestive tract in a delayed manner. Examples of embedding components that can be used are polymeric substances and wax substances. In addition, the active ingredient can also be formed into a microcapsule form with one or more of the above-mentioned carriers.

In another embodiment of the present invention, the above-mentioned pharmaceutical preparation is a liquid dosage form for oral administration, including (but not limited to) pharmaceutically acceptable emulsions, solutions, suspensions, syrups, tinctures, etc.

In another embodiment of the present invention, the above-mentioned pharmaceutical preparation is a dosage form for parenteral injection, including (but not limited to) physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions and dispersions.

In another embodiment of the present invention, the above-mentioned pharmaceutical preparation is a dosage form for topical administration, including (but not limited to) ointments, powders, suppositories, drops, sprays, inhalants and the like.

[Medical Use]

Whether it is the compound of the present invention or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope label or prodrug, or the pharmaceutical composition of the present invention, or the pharmaceutical preparation of the present invention, it can show inhibitory activity on SOS1. Therefore, the present invention provides the use of the compound of the present invention or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope label or prodrug, or the pharmaceutical composition of the present invention, or the pharmaceutical preparation of the present invention in the preparation of a medicament for preventing and/or treating a disease mediated at least in part by the SOS1 protein.

The present invention provides use of the compound of the present invention or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, or the pharmaceutical composition of the present invention or the pharmaceutical preparation of the present invention in preparing a drug for preventing and/or treating cancer.

The term "cancer" refers to a cellular disorder characterized by uncontrolled or dysregulated cell proliferation, reduced cell differentiation, inappropriate ability to invade surrounding tissues, and/or the ability to establish new growth at ectopic sites. Non-limiting examples of cancer include, but are not limited to, pancreatic cancer, lung cancer, colorectal cancer, bile duct cancer, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial carcinoma, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B-cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular carcinoma, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer, and sarcoma.

The present invention also provides a compound of the present invention or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or a pharmaceutical composition of the present invention or a pharmaceutical preparation of the present invention, which is used for preventing and/or treating a disease (especially cancer) mediated at least in part by SOS1 protein.

The present invention also provides a method for preventing and/or treating a disease (especially cancer) at least partially mediated by SOS1 protein, comprising administering a preventive and/or therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, or a pharmaceutical composition of the present invention, or a pharmaceutical preparation of the present invention to an individual in need thereof.

In one embodiment of the present invention, the cancer is pancreatic cancer, non-small cell lung cancer, or colorectal cancer.

The technical solutions of the present invention will be further described below in conjunction with specific embodiments. Unless otherwise specified, the following embodiments use The reagents, materials, instruments, etc. can all be obtained through conventional commercial means, and the experimental methods used are conventional methods in the art.

In the following examples, the following abbreviations are used:

THF             tetrahydrofuran

PE Petroleum ether

EA              Ethyl acetate

Diox              1,4-Dioxane

MeOH           Methanol

EtOH

AcOH            acetic acid

DMF             N,N-Dimethylformamide

DIPEA           N,N-Diisopropylethylamine

TEA             Triethylamine

DCM            Dichloromethane

DCE              1,2-Dichloroethane

DMAP            4-Dimethylaminopyridine

Boc              tert-Butyloxycarbonyl

Pyr

Tol              Toluene

MeCN            Acetonitrile

TFA             Trifluoroacetic acid

NBS              N-Bromosuccinimide

Comparative Example 1: Preparation of positive control compound 1

(1) Synthesis route of compound 1-1

Synthesis of compound 1-1b:

Under nitrogen protection, compound 1-1a (20.00 g, 106.36 mmol, 1.0 eq), THF (150 mL), (R)-tert-butylsulfenamide (19.31 g, 159.54 mmol, 1.5 eq) and tetraethyl titanate (74.65 g, 327.26 mmol, 2.0 eq) were added to the reaction bottle in sequence, and the temperature was raised to 80°C and stirred for 4 h. After the reaction system was cooled to room temperature, water (150 mL) was added, and EA was used for extraction (150 mL*2). The organic phase was dried over anhydrous Na ₂ SO ₄ , concentrated, and purified by column chromatography (eluent: PE/EA=5/1, v/v) to obtain compound 1-1b (28.0 g); MS: m/z 292.1[M+1] ⁺ ;

Synthesis of compound 1-1c:

Compound 1-1b (28.00 g, 96.19 mmol, 1.0 eq), THF (400 mL) and water (6 mL) were added to the reaction flask in sequence. After cooling to -60 to -50 °C, sodium borohydride (6.58 g, 173.14 mmol, 1.8 eq) was slowly added. After the addition, the temperature was raised to -5 to 0 °C and the stirring was continued for 30 min. Water (300 mL) was added to the system to quench the reaction, and EA was added for extraction (300 mL*2). The organic phase was dried over anhydrous Na ₂ SO ₄ , concentrated, and purified by column chromatography (eluent: PE/EA=5/1, v/v) to obtain compound 1-1c (22.2 g); MS: m/z 294.1[M+1] ⁺ ;

Synthesis of compound 1-1:

Compound 1-1c (22.20 g, 75.76 mmol, 1.0 eq), Diox (50 mL) and 4 M HCl (40 mL) were added to the reaction flask in sequence, and the mixture was stirred at room temperature for 2 h. The pH value of the system was adjusted to 8-9 using 1 M NaOH aqueous solution, and extracted with EA (100 mL*2). The organic phase was dried with anhydrous Na ₂ SO ₄ and concentrated. The obtained solid was slurried with PE to obtain compound 1-1 (10.0 g); MS: m/z 190.1[M+1] ⁺ .

(2) Synthesis route of compound 1-2

Synthesis of compound 1-2b:

At 0°C, concentrated H ₂ SO ₄ (160 mL) and compound 1-2a (20.0 g, 91.76 mmol, 1.0 eq) were added to the reaction flask in sequence, and KNO ₃ (9.27 g, 91.76 mmol, 1.0 eq) was slowly added in batches. The reaction system was reacted at room temperature for 1 h. After the reaction was completed, the system was slowly poured into ice water (500 mL) and stirred. After returning to room temperature, it was filtered, and the filter cake was rinsed with water twice and dried to obtain compound 1-2b (25.00 g), yield: 100%; MS: m/z 261.8[M-1] ⁺ ;

Synthesis of compound 1-2c:

Compound 1-2b (25.00 g, 95.08 mmol, 1.0 eq), MeOH (400 mL) and concentrated H ₂ SO ₄ (4.26 g, 120.41 mmol, 1.2 eq) were added to the reaction flask in sequence. The reaction system was heated to 70°C. After reacting for 16 h, the system was cooled to room temperature, concentrated to remove most of the solvent, and then filtered. The obtained solid was rinsed twice with water and dried to obtain compound 1-2c (18.00 g). Yield: 71.2%; MS: m/z 277.8 [M+1] ⁺ ;

Synthesis of compound 1-2d:

Compound 1-2c (10.00 g, 36.11 mmol, 1.0 eq), DMF (100 mL), DL-alanine methyl ester hydrochloride (7.56 g, 54.16 mmol, 1.5 eq) and TEA (9.14 g, 90.28 mmol, 2.5 eq) were added to the reaction flask in sequence, the reaction system was heated to 80°C, reacted for 2 h, then the system was cooled to room temperature, water (300 mL) was added and stirred, filtered, the filter cake was rinsed twice with water, and dried to obtain compound 1-2d (12.50 g), yield: 96.9%; MS: m/z 360.8[M+1] ⁺ ;

Synthesis of compound 1-2e:

Compound 1-2d (12.50 g, 34.72 mmol, 1.0 eq), anhydrous EtOH (150 mL) and SnCl ₂ (32.90 g, 173.6 mmol, 5.0 eq) were added to the reaction flask in sequence. The reaction system was heated to 95°C. After reacting for 3 h, the system was cooled to room temperature. A saturated aqueous solution of NaHCO ₃ was added to adjust the pH value to neutral. EA (150 mL) was added and stirred evenly, and then filtered. The filter cake was rinsed with EA (200 mL*3), and the filtrate was separated. The organic phase was washed with water and saturated brine in sequence, and then dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to obtain compound 1-2e (6.10 g). Yield: 59.2%; MS: m/z 299.0[M+1] ⁺ ;

Synthesis of compound 1-2f:

At 0°C, compound 1-2e (6.10 g, 20.47 mmol, 1.0 eq) and DMF (110 mL) were added to the reaction bottle in sequence, and then NaH (4.09 g, 102.35 mmol, 5.0 eq) was slowly added in batches. After stirring for 20 min, CH ₃ I (311.6 g, 81.88 mmol, 4.0 eq) was added. After reacting for 30 min, water (150 mL) was added to the system to quench the reaction, and EA (150 mL*3) was added for extraction. The organic layers were combined, washed with water and saturated brine in sequence, dried with anhydrous Na ₂ SO ₄ , filtered, concentrated, and purified by column chromatography (eluent: PE/EA=3:1, v/v) to obtain compound 1-2f (2.50 g), yield: 37.5%; MS: m/z 327.0[M+1] ⁺ ;

Synthesis of compound 1-2h:

Under nitrogen protection, compound 1-2f (2.50 g, 7.67 mmol, 1.0 eq), Diox (30 mL), tributyl (1-ethoxyethylene) tin (4.15 g, 11.5 mmol, 1.5 eq), Pd (dppf) ₂ Cl ₂ (0.84 g, 1.15 mmol, 0.15 eq) and TEA (2.32 g, 23.01 mmol, 3.0 eq) were added to the reaction bottle in sequence, and the reaction system was heated to 90 ° C. After reacting for 7 hours, the system was cooled to room temperature, and dilute HCl was added to adjust the pH value to weak acidity. The reaction product was monitored by TLC and HR-MS to be converted from compound 1-2g to compound 1-2h, and EA was added for extraction (50 mL * 3). The organic layers were combined, washed with water and saturated brine in sequence, dried by adding anhydrous Na ₂ SO ₄ , filtered, concentrated, and purified by column chromatography (washing Stripping agent: PE/EA = 1:1, v/v), to obtain compound 1-2h (1.79 g), yield: 80.6%; MS: m/z 291.2 [M+1] ⁺ ;

Synthesis of compound 1-2i:

Compound 1-2h (1.79 g, 6.17 mmol, 1.0 eq), anhydrous EtOH (20 mL) and hydrazine hydrate (1.54 g, 24.68 mmol, 4.0 eq) were added to the reaction flask in sequence. The reaction system was heated to 95°C. After reacting for 2 h, the system was cooled to room temperature, concentrated to remove most of the solvent and then filtered. The obtained solid was dried to obtain compound 1-2i (1.50 g). Yield: 89.8%; MS: m/z 273.0 [M+1] ⁺ .

Synthesis of compound 1-2:

Compound 1-2i (1.50 g, 5.51 mmol, 1.0 eq), POCl ₃ (20 mL) and N,N-diethylaniline (1.23 g, 8.26 mmol, 1.5 eq) were added to the reaction flask in sequence. The reaction system was heated to 108°C. After reacting for 2 h, the system was concentrated under low pressure to remove the solvent, ice water (20 mL) was added, and the pH value was adjusted to 7-8 with a saturated aqueous NaHCO ₃ solution. EA was added for extraction (30 mL*3), the organic layers were combined, washed once with water and saturated brine in sequence, dried with anhydrous Na ₂ SO ₄ , filtered, concentrated, and purified by column chromatography (eluent: PE/EA=1:2, v/v) to obtain compound 1-2 (1.21 g), yield: 75.6%; MS: m/z 291.0[M+1] ⁺ .

(3) Synthesis route of positive control compound 1

Under nitrogen protection, compound 1-2 (300.0 mg, 1.03 mmol, 1.0 eq), Tol (6 mL), compound 1-1 (255.9 mg, 2.07 mmol, 2.0 eq), Pd(dba) ₃ (372.1 mg, 0.62 mmol, 0.6 eq), Xantphos (470.2 mg, 1.24 mmol, 1-2 eq) and K ₃ PO ₄ (862.5 mg, 6.20 mmol, 6.0 eq) were added to a microwave tube in sequence, and the reaction was carried out at 110° C. for 2 h. The product was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=20/1, v/v) to obtain positive control compound 1 (83.21 mg) with a yield of 18.3%; MS: m/z 444.2[M+1] ⁺ .

Example 1: Preparation of Compound 1

Except that compound 1-1 was replaced by compound 1-5, the other operations were basically similar to those of positive control compound 1 to synthesize compound 1 (29.5 mg) with a yield of 54%; MS: m/z 415.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ8.57 (s, 1H), 8.23 (d, J=10.2Hz, 1H), 7.88 (t, J=8.7Hz, 1H), 7.66-7.63 (m, 1H), 7.35 (q, J=8.0Hz, 1H), 7.04 (d, J=4.3Hz, 1H), 5.49 (p, J=6 .9Hz, 1H), 4.39-4.29 (m, 1H), 3.57 (d, J=5.1Hz, 3H), 3.07 (d, J=1.6Hz, 3H), 2.72 (s, 3H), 2.69 (s, 3H), 1.69-1.56 (m, 3H), 1.18-1.15 (m, 3H).

Example 2: Preparation of Compound 2

Except for replacing compound 1-1 with compound 2-1, the other operations were basically similar to those of the positive control compound 1 to synthesize compound 2 (18.8 mg) with a yield of 21%; MS: m/z 474.4 [M+1] ⁺ .

Example 3: Preparation of Compound 3

Except that iodomethane was replaced by perdeuterated iodomethane, the other operations were basically similar to those of compound 1-2 to synthesize compound 3-1 (0.88 g) with a yield of 69%; MS: m/z 297.1 [M+1] ⁺ . Except that compound 1-2 was replaced by compound 3-1, the other operations were basically similar to those of positive control compound 1 to synthesize compound 3 (8.8 mg) with a yield of 23%; MS: m/z 450.4 [M+1] ⁺ .

Example 4: Preparation of Compound 4

Except that DL-alanine methyl ester hydrochloride was replaced by 2-aminoisobutyric acid methyl ester, the other operations were basically similar to those of compound 1-2 to synthesize compound 4-1 (0.58 g) with a yield of 71%; MS: m/z 305.1 [M+1] ⁺ . Except that compound 1-2 was replaced by compound 4-1, the other operations were basically similar to those of positive control compound 1 to synthesize compound 4 (18.3 mg) with a yield of 53%; MS: m/z 429.4 [M+1] ⁺ .

Example 5: Preparation of Compound 5

Compound 5 (23.8 mg) was synthesized in a yield of 22%, except that compound 1-2 was replaced by compound 4-1 and compound ^{1-1 was} replaced by compound 2-1 _. ): δ7.82 (t, J=7.0Hz, 1H), 7.63 (t, J=7.5Hz, 1H), 7.38 (t, J=7.8Hz, 1H), 7.23 (s, 1H), 7.07 (s, 1H), 5.22-5.1 1 (m, 2H), 4.38 (q, J=6.6Hz, 1H), 3.37 (s, 3H), 3.01 (s, 3H), 2.74 (s, 3H), 1.37 (s, 6H), 1.27 (d, J=6.7Hz, 3H).

Example 6: Preparation of Compound 6

Under nitrogen protection, compound 4-1 (100.0 mg, 0.34 mmol, 1.0 eq), Tol (6 mL), compound 1-3 (227.7 mg, 0.66 mmol, 2.0 eq), Pd(dba) ₃ (180.7 mg, 0.21 mmol, 0.6 eq), Xantphos (228.4 mg, 41.3 mmol, 1.2 eq) and K ₃ PO ₄ (418.9 mg, 2.07 mmol, 6.0 eq) were added to a microwave tube in sequence, and the mixture was reacted at 110° C. for 2 h. The mixture was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=20/1, v/v) to obtain compound 6a (35.11 mg, 0.06 mmol). DCM (5 mL) was added as solvent, TFA (0.2 mL) was added, and the mixture was stirred at room temperature for 10 min. NaHCO ₃ The aqueous solution was adjusted to pH 8, extracted with DCM (10 mL*3), the organic phases were combined, concentrated in vacuo, and purified by column chromatography (EA/MeOH=4/1, v/v) to give compound 6 (2.35 mg), yield: 7.99%; MS: m/z 515.3 [M+1] ⁺ .

Example 7: Preparation of Compound 7

Under nitrogen protection, compound 4-1 (100.0 mg, 0.34 mmol, 1.0 eq), Tol (6 mL), compound 1-4 (171.1 mg, 0.66 mmol, 2.0 eq), Pd(dba) ₃ (180.7 mg, 0.21 mmol, 0.6 eq), Xantphos (228.4 mg, 41.3 mmol, 1.2 eq) and K ₃ PO ₄ (418.9 mg, 2.07 mmol, 6.0 eq) were added into a microwave tube in sequence, and the mixture was reacted at 110° C. for 2 h. The mixture was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=4/1, v/v) to obtain compound 7 (5.11 mg). The yield was 2.94%; MS: m/z 529.4[M+1] ⁺ .

Example 8: Preparation of Compound 8

(1) Synthesis route of compound 8-1

Synthesis of compound 8-1a:

Compound 1-2e (4.00 g, 13.42 mmol, 1.0 eq), (Boc) ₂ O (3.80 g, 17.45 mmol, 1.3 eq), DMAP (655.9 mg, 5.37 mmol, 0.4 eq) and DCM (40 mL) were added to the reaction bottle in sequence, and the reaction was allowed to proceed overnight at room temperature. The reaction system was poured into water (100 mL), and extracted with DCM (120 mL*3). The organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=5/14, v/v) to obtain compound 8-1a (3.44 g), with a yield of 64.4%; MS: m/z 343.2 [M- ^t Bu+1] ⁺ ;

Synthesis of compound 8-1b:

Compound 8-1a (1.87 g, 4.70 mmol, 1.0 eq), cyclopropylboronic acid (1.21 _g , 14.10 mmol, 3.0 eq), Cu(OAc) ₂ (0.85 g, 4.70 mmol, 1.0 eq), Pyr (2.23 g, 28.20 mmol, 6.0 eq) and MeCN (20 mL) were added to the reaction bottle in sequence, and the mixture was reacted at 60°C for 40 h. The reaction system was poured into cold water (20 mL), extracted with EA (50 mL*3), the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=3/1, v/v) to obtain compound 8-1b (1.70 g), yield: 82.9%; MS: m/z 383.2[M- ^t Bu+1] ⁺ ;

Synthesis of compound 8-1c:

Compound 8-1b (1.70 g, 3.88 mmol, 1.0 eq), DCM (35 mL) and TFA (5 mL) were added to the reaction bottle in sequence, stirred at room temperature for 2 h, the reaction system was spin-dried, redissolved with DCM (15 mL), the pH value was adjusted to 8 with saturated NaHCO ₃ aqueous solution, extracted with DCM (20 mL*3), the organic phases were combined, dried with anhydrous Na ₂ SO ₄ , filtered, and spin-dried to obtain compound 8-1c (1.29 g), yield: 98.5%; MS: m/z 339.1[M+1] ⁺ ;

The synthesis operation of compound 8-1d to compound 8-1 is basically the same as the synthesis operation of compound 1-2e to compound 1-2 in Comparative Example 1.

Compound 8-1d (yield 90.5%), MS: m/z 353.0 [M+1] ⁺ ; Compound 8-1e (yield 81.2%), MS: m/z 345.2 [M+1] ⁺ ; Compound 8-1f (yield 85.1%), MS: m/z 317.1 [M+1] ⁺ ; Compound 8-1g (yield 91.2%), MS: m/z 353.0 [M+1] ⁺ ; Compound 8-1 (yield 73.5%), MS: m/z 317.1 [M+1] ⁺ ;

(2) Synthesis route of compound 8

Under nitrogen protection, compound 8-1 (90.0 mg, 0.28 mmol, 1.0 eq), Tol (6 mL), compound 1-5 (91.0 mg, 0.56 mmol, 2.0 eq), Pd(dba) ₃ (155.7 mg, 0.17 mmol, 0.6 eq), Xantphos (198.0 mg, 0.34 mmol, 1.2 eq) and K ₃ PO ₄ (362.7 mg, 1.68 mmol, 6.0 eq), microwave 110° C. for 2 h, filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=20/1, v/v) to give compound 8 (20.21 mg), yield: 16.2%; MS: m/z 441.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ8.73(d, J=15.2Hz, 1H), 8.44-8.39(m, 1H), 8.05-7.94(m, 1H), 7.87-7.67(m , 1H), 7.48 (s, 1H), 6-27-6.17 (m, 1H), 4.63-4.52 (m, 1H), 3.35-3.28 (m, 1H), 3.2 2(d, J=3.8Hz, 3H), 3.12 (d, J=2.5Hz, 3H), 2.56 (d, J=7.0Hz, 2H), 2.09-2.03 (m, 3 H), 1.81-1.70 (m, 3H), 1.67-1.60 (m, 4H), 1.45-1.34 (m, 1H), 0.91-0.84 (m, 1H).

Example 9: Preparation of Compound 9

(1) Synthesis route of compound 9-1

Synthesis of compound 9-1a:

Compound 1-2c (10.00 g, 36.11 mmol, 1.0 eq), DCE (50 mL), and cyclopropylamine (4.12 g, 72.22 mmol, 2.0 eq) were added to the reaction flask in sequence. The reaction system was heated to 80°C. After reacting for 2 h, the system was cooled to room temperature, water (100 mL) was added and stirred, and the mixture was extracted with DCM (150 mL*3). The mixture was dried over anhydrous Na ₂ SO ₄ , filtered, and concentrated to obtain compound 9-1a (10.2 g). Yield: 89.5%; MS: m/z 315.1[M+1] ⁺ ;

Synthesis of compound 9-1b:

Compound 9-1a (10.0 g, 31.85 mmol, 1.0 eq), anhydrous EtOH (100 mL), and anhydrous SnCl ₂ (18.11 g, 95.54 mmol, 3 eq) were added to the reaction flask in sequence, and the mixture was reacted at 90°C for 2 h. The mixture was cooled to room temperature, and the reaction system was poured into water/EA (1000 mL/1000 mL) and stirred. The mixture was filtered, and the filter cake was rinsed with EA for 3 times. The liquid phase was retained and extracted with EA (150 mL*5). The organic phases were combined, dried with anhydrous Na ₂ SO ₄ , filtered, and concentrated in vacuo to obtain compound 9-1b (5.20 g). Yield: 57.5%; MS: m/z 285.1[M+1] ⁺ ;

Synthesis of compound 9-1c:

Under nitrogen protection, compound 9-1b (5.2 g, 18.31 mmol, 1.0 eq) and DIPEA (8.5 mL, 36.62 mmol, 2 eq) were added to the reaction bottle in sequence, and anhydrous DCM (60 mL) was used as solvent. Then, the mixture was stirred under an ice bath, and 2-bromopropionyl bromide (3.74 g, 27.46 mmol, 1.5 eq) was added. After the addition, the mixture was moved to room temperature for reaction for 2 h. The reaction system was poured into cold water (100 mL), extracted with DCM (100*3), the organic phases were combined, dried with anhydrous Na ₂ SO ₄ , filtered, and spin-dried. Then, MeCN (20 mL) was added, and DIPEA (9 mL) was added and the temperature was raised to 80°C for reaction overnight. After the reaction was completed, the reaction system was poured into water (30 mL), extracted with DCM (50 mL*3), the organic phases were combined, and the organic phases were purified by anhydrous Na ₂ SO ₄ Drying, filtration, concentration, and column chromatography purification (PE/EA=1/1, v/v) to obtain compound 9-1c (2.9 g), yield: 46.8%; MS: m/z 339.1 [M+1] ⁺ ;

The synthesis operation of compound 9-1d to compound 9-1 is basically the same as the synthesis operation of compound 1-2e to compound 1-2 in Comparative Example 1.

Compound 9-1d (yield 88.2%), MS: m/z 353.0 [M+1] ⁺ ; Compound 9-1e (yield 82.8%), MS: m/z 316.1 [M+1] ⁺ ; Compound 9-1f (yield 86.4%), MS: m/z 299.1 [M+1] ⁺ ; Compound 9-1 (yield 75.6%), MS: m/z 317.1 [M+1] ⁺ ;

(2) Synthesis route of compound 9

Under nitrogen protection, compound 9-1 (100.0 mg, 0.32 mmol, 1.0 eq), Tol (6 mL), compound 1-5 (101.0 mg, 0.64 mmol, 2.0 eq), Pd(dba) ₃ (172.8 mg, 0.20 mmol, 0.6 eq), Xantphos (219.8 mg, 0.38 mmol, 1.2 eq) and K ₃ PO ₄ (398.9 mg, 1.92 mmol, 6.0 eq) were added into a microwave tube in sequence, and the reaction was carried out at 110° C. for 2 h. The product was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=20/1, v/v) to obtain compound 9 (22.11 mg). The yield was 15.9%; MS: m/z 441.4[M+1] ⁺ .

Example 10: Preparation of Compound 10

(1) Synthesis route of compound 10-1

Synthesis of compound 10-1a:

The synthesis operation of compound 1-2d in Comparative Example 1 was basically the same to obtain compound 10-1a (5.84 g) with a yield of 95%; MS: m/z 373.1 [M+1] ⁺ ;

Synthesis of compound 10-1b:

Compound 10-1a (3.20 g, 8.60 mmol, 1.0 eq) was added to a reaction bottle, and iron powder (1.45 g, 3 eq) was added, and AcOH (30 mL) was used as solvent. The reaction was carried out at 90°C for 1 h, and the reaction system was poured into cold water (100 mL). The pH value was adjusted to 8 with potassium carbonate, and extracted with EA (100 mL*3). The organic phases were combined, washed three times with saturated brine, dried with anhydrous Na ₂ SO ₄ , filtered, and concentrated to obtain compound 10-1b (2.80 g), yield: 100%; MS: m/z 311.2[M+1] ⁺ ;

The synthesis operation of compound 10-1c to compound 10-1 is basically the same as the synthesis operation of compound 1-2e to compound 1-2 in Comparative Example 1.

10-1c (yield 82.4%), MS: m/z 339.0 [M+1] ⁺ ; 10-1d (yield 86.5%), MS: m/z 303.1 [M+1] ⁺ ; 10-1e (yield 83.5%), MS: m/z 285.1 [M+1] ⁺ ; 10-1 (yield 77.8%), MS: m/z 303.1 [M+1] ⁺ ;

(2) Synthesis route of compound 10

Under nitrogen protection, compound 10-1 (100.0 mg, 0.33 mmol, 1.0 eq), Tol (6 mL), compound 1-5 (106.0 mg, 0.66 mmol, 2.0 eq), Pd(dba) ₃ (181.9 mg, 0.20 mmol, 0.6 eq), Xantphos (229.8 mg, 0.40 mmol, 1.2 eq) and K ₃ PO ₄ (412.6 mg, 1.98 mmol, 6.0 eq) were added to a microwave tube in sequence, and the mixture was reacted at 110° C. for 2 h. The mixture was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=20/1, v/v) to obtain compound 10 (55.01 mg), with a yield of 39.1%; MS: m/z 427.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ7.91 (s, 1H), 7.78 (d, J=7.9Hz, 1H), 7.62 (d, J=7.6Hz, 2H), 7.33 (t, J=7.8Hz, 1H), 7.00 (s, 1H), 5.59 (t, J=7.0Hz, 1 H), 3.52 (s, 3H), 2.82 (s, 3H), 2.69 (s, 3H), 2.60 (s, 3H), 1.58 (d, J=7.0Hz, 3H), 1.45-1.31 (m, 2H), 1.29-1.08 (m, 2H).

Example 11: Preparation of Compound 11

(1) Synthesis route of compound 11-1

The synthesis operation of compound 11-1a to compound 11-1 is basically the same as the synthesis operation of compound 1-2f to compound 1-2 in Comparative Example 1.

Compound 11-1a (yield 89.7%), MS: m/z 413.0 [M+1] ⁺ ; Compound 11-1b (yield 85.5%), MS: m/z 405.2 [M+1] ⁺ ; Compound 11-1c (yield 84.7%), MS: m/z 377.1 [M+1] ⁺ ; Compound 11-1d (yield 65.7%), MS: m/z 359.2 [M+1] ⁺ ; Compound 11-1 (yield 77.4%), MS: m/z 277.1 [M+1] ⁺ ;

(2) Synthesis route of compound 11

Synthesis of compound 11-1e:

Under nitrogen protection, compound 11-1 (100.0 mg, 0.36 mmol, 1.0 eq), Tol (6 mL), compound 1-5 (116.0 mg, 0.72 mmol, 2.0 eq), Pd(dba) ₃ (199.0 mg, 0.22 mmol, 0.6 eq), Xantphos (251.6 mg, 0.43 mmol, 1.2 eq) and K ₃ PO ₄ (4461.3 mg, 2.16 mmol, 6.0 eq) were added to a microwave tube in sequence, and the mixture was reacted at 110° C. for 2 h. The mixture was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=10/1, v/v) to obtain compound 11-1e (23.11 mg), with a yield of 15.9%; MS: m/z 401.4[M+1] ⁺ ;

Synthesis of compound 11:

Compound 11-1e (23.0 mg, 0.06 mmol, 1 eq), MnO ₂ (50.0 mg, 0.6 mmol, 10 eq) and DCM (5 mL) were added to the reaction bottle, stirred at room temperature for 1 h, filtered through celite, rinsed with DCM (30 mL), and the filtrate was dried by spin drying. The product was purified by column chromatography (EA/MeOH=10/1, v/v) to obtain compound 11 (7.35 mg), yield: 32.1%; MS: m/z 399.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ8.36 (s, 1H), 8.31 (s, 1H), 7.87 (d, J = 6.6Hz, 1H), 7.80 (d, J = 7.9Hz, 1H), 7.63 (d, J = 7.7Hz, 1H), 7.34 (t, J =7.7Hz, 1H), 5.67-5.63 (m, 1H), 3.80 (s, 3H), 2.71 (s, 3H), 2.69 (s, 3H), 2.54 (s, 3H), 1.62 (d, J = 7.0Hz, 3H).

Comparative Example 2: Preparation of positive control compound 2

Except for replacing compound 1-5 with compound 1-1, the other operations were basically similar to compound 11 to synthesize positive control compound 2 (39.5 mg) with a yield of 24%; MS: m/z 428.2 [M+1] ⁺ .

Example 12: Preparation of Compound 12

(1) Synthesis route of compound 12-1

Synthesis of compound 12-1b:

At 10°C, compound 12-1a (20.0 g, 114.2 mmol, 1.0 eq) and AcOH (150 mL) were added to the reaction bottle, and then sodium cyanoborohydride (32.6 g, 342.7 mmol, 3 eq) was slowly added in batches. After the addition, the mixture was moved to room temperature for 2 h. The reaction system was poured into ice water (500 mL), and the pH value was adjusted to 8 with solid sodium carbonate. EA (300 mL*3 extractions) was used, and the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=3/1, v/v) to obtain compound 12-1b (11.3 g), with a yield of 55.9%; MS: m/z 178.2[M+1] ⁺ ;

Synthesis of compound 12-1c:

Compound 12-1b (7.00 g, 39.5 mmol, 1.0 eq), tetrahydrofuran-3(2H)-one (6.81 g, 779.1 mmol, 2 eq), MeOH (80 mL) and AcOH (2 mL) were added to the reaction flask in sequence, and sodium triacetoxyborohydride (22.43 g, 118.6 mmol, 3 eq) was slowly added in batches at 0°C-10°C. After the addition, the temperature was naturally raised to room temperature. The reaction was allowed to react overnight. The reaction system was spin-dried, diluted with water (100 mL), extracted with EA (150 mL*3), the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=5/1, v/v) to obtain compound 12-1c (4.16 g), yield: 42.6%; MS: m/z 248.2[M+1] ⁺ ;

Synthesis of compound 12-1d:

Compound 12-1c (4.1 g, 16.6 mmol, 1.0 eq) and DMF (40 mL) were added to the reaction flask, and NBS (3.25 g, 18.3 mmol, 1.1 eq) was slowly added under an ice bath. After the addition, the mixture was moved to room temperature for reaction for 2 h. The reaction system was poured into cold water (50 mL), extracted with EA (50 mL*3), the organic phases were combined, washed with saturated sodium chloride aqueous solution (60 mL*3), dried over anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=2/1, v/v) to obtain compound 12-1d (3.9 g), yield: 72.4%; MS: m/z 326.0[M+1] ⁺ ;

The synthesis operation of compound 12-1e to compound 12-1 is basically the same as the synthesis operation of compound 1-2g to compound 1-2 in Comparative Example 1.

Compound 12-1e (yield 74.4%), MS: m/z 318.2 [M+1] ⁺ ; Compound 12-1f (yield 85.3%), MS: m/z 290.1 [M+1] ⁺ ; Compound 12-1g (yield 84.5%), MS: m/z 272.1 [M+1] ⁺ ; Compound 12-1 (yield 77.4%), MS: m/z 290.1 [M+1] ⁺ ;

(2) Synthesis route of compound 12

Under nitrogen protection, compound 12-1 (30.0 mg, 0.11 mmol, 1.0 eq), Tol (6 mL), compound 1-1 (39.2 mg, 0.22 mmol, 2.0 eq), Pd(dba) ₃ (57.03 mg, 0.07 mmol, 0.6 eq), Xantphos (72.1 mg, 0.13 mmol, 1.2 eq) and K ₃ PO ₄ (132.2 mg, 0.65 mmol, 6.0 eq) were added to a microwave tube in sequence, and the mixture was reacted at 110° C. for 2 h. The mixture was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=10/1, v/v) to obtain compound 12 (2.00 mg), with a yield of 4.4%; MS: m/z 443.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ7.75 (s, 1H), 7.70-7.66 (m, 2H), 7.58 (s, 1H), 7.48 (t, J=7.1Hz, 1H), 7.26 ( t, J=8.2Hz, 1H), 7.25 (t, J=54.1Hz, 1H), 5.66-5.59 (m, 1H), 4.80-4.74 (m, 1H) , 4.01-3.93(m, 1H), 3.92-3.83(m, 2H), 3.81-3.65(m, 3H), 3.18(t, J=8.3Hz, 2 H), 2.57 (s, 3H), 2.40-2.29 (m, 1H), 2.10-1.99 (m, 1H), 1.66 (d, J=7.0Hz, 3H).

Example 13: Preparation of Compound 13

Except that compound 1-1 was replaced by compound 1-5, the rest was synthesized by the same operation as compound 12 (19.9 mg), with a yield of 51%; MS: m/z 414.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ7.94 (s, 1H), 7.80 (d, J=7.9 Hz, 1H), 7.74 (s, 1H), 7.64 (d, J=7.6 Hz, 1H), 7.55 (s, 1H), 7.35 (t, J=7.8 Hz, 1H), 5.44-5.38 (m, 1H), 4.82-4.77 (m, 1H), 3.99 (q, J=7.7 Hz, 1H); .7Hz, 1H), 3.94-3.84(m, 2H), 3.78(q, J＝8.5Hz, 3H), 3.21(t, J＝8.3Hz, 2H), 2.69( s, 3H), 2.61 (s, 3H), 2.40-2.30 (m, 1H), 2.09-2.04 (m, 1H), 1.61 (d, J=6.9Hz, 3H).

Example 14: Preparation of Compound 14

Compound 13 (25.0 mg, 0.06 mmol, 1 eq), MnO ₂ (50.0 mg, 0.6 mmol, 10 eq) and DCM (5 mL) were added to the reaction bottle, stirred at room temperature for 1 h, filtered through celite, rinsed with DCM (30 mL), and the filtrate was dried by spin drying. The product was purified by column chromatography (EA/MeOH=10/1, v/v) to give compound 14 (10.66 mg), yield: 42.8%; MS: m/z 412.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ8.95 (s, 1H), 8.37 (s, 1H), 7.98-7.94 (m, 2H), 7.86 (d, J=8.0, 1H), 7.61 ( d, J=7.6Hz, 1H), 7.32 (t, J=7.8Hz, 1H), 6.90 (d, J=3.3Hz, 1H), 5.61 (t, J=6.5 Hz, 1H), 5.575.49 (m, 1H), 4.22-4.10 (m, 2H), 4.03-3.91 (m, 2H), 2.75 (s, 3H ), 2.70 (s, 3H), 2.69-2.61 (m, 1H), 2.30-2.19 (m, 1H), 1.64 (d, J=6.9Hz, 3H).

Example 15: Preparation of Compound 15

Compound 15 (9.9 mg) was synthesized by the same operation as compound 14, except that compound 13 was replaced by compound 12. The yield was 43%. MS: m/z 441.4 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ9.17 (s, 1H), 8.41 (s, 1H), 8.21 (s, 1H), 7.97 (dd, J=3.4, 1.2 Hz, 1H), 7.76 (t, J=7.7 Hz, 1H), 7.49 (t, J=7.1 Hz, 1H), 7.27 (s, 1H), 7.25 (t, J=54.2 Hz, 1H), 6.92 ( d, J=3.3Hz, 1H), 5.78-5.71 (m, 1H), 5.64-5.58 (m, 1H), 4.20-4.11 (m, 2H), 4.04- 3.88 (m, 2H), 2.77 (s, 3H), 2.74-2.61 (m, 1H), 2.26 (s, 1H), 1.73 (d, J=7.0Hz, 3H).

Example 16: Preparation of Compound 16

(1) Synthesis route of compound 16-1

Synthesis of compound 16-1b:

Compound 16-1a (5.0 g, 26.2 mmol, 1.0 eq), CH ₃ I (7.43 g, 52.4 mmol, 2.0 eq) and DMF (100 mL) were added to the reaction flask, the reaction system was cooled to 0°C, 60% NaH (2.10 g, 25.4 mmol, 2.0 eq) was slowly added in batches, and the reaction was maintained at 0°C for 1 h. The reaction solution was added to ice water (200 mL), extracted with EA (200 mL*3), the organic phases were combined, washed with saturated sodium chloride aqueous solution (300 mL*3), dried with anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=2/1, v/v) to obtain compound 16-1b (5.1 g), yield: 89.1%; MS: m/z 220.0[M+1] ⁺ ;

Synthesis of compound 16-1c:

Under nitrogen protection, compound 16-1b (5.0 g, 22.8 mmol, 1.0 eq) and anhydrous THF (100 mL) were added to the reaction bottle, the system temperature was lowered to -15 °C, sodium borohydride (2.6 g, 68.7 mmol, 3 eq) was slowly added in batches, and then boron trifluoride ether solution (11.3 g, 79.6 mmol, 3.5 eq) was slowly added to the system, and the temperature was raised to room temperature for 4 h after the addition. The reaction system was lowered to 0 °C, the pH value was adjusted to 4 with 1M HCl, stirred at room temperature for 2 h, extracted with EA (150 mL*3), the organic phases were combined, dried with anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=2/1, v/v) to obtain compound 16-1c (3.88 g), yield: 82.9%; MS: m/z 206.1[M+1] ⁺ ;

Synthesis of compound 16-1d:

Compound 16-1c (3.8 g, 18.5 mmol, 1.0 eq) and MeCN (50 mL) were added to the reaction flask, and NBS (3.3 g, 18.5 mmol, 1.0 eq) was slowly added at -30 °C. After reacting for 0.5 h, the reaction system was poured into cold water (50 mL), extracted with EA (50 mL*3), the organic phases were combined, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=5/1, v/v) to obtain compound 16-1d (3.0 g), yield: 72.4%; MS: m/z 283.8[M+1] ⁺ ;

Synthesis of compound 16-1e:

Compound 16-1d (3.0 g, 10.5 mmol, 1.0 eq), tetrahydrofuran-3(2H)-one (1.82 g, 21.1 mmol, 2 eq), MeOH (40 mL) and AcOH (0.5 mL) were added to the reaction bottle in sequence, and sodium cyanoborohydride (2.6 g, 41.99 mmol, 4 eq) was slowly added in batches at 0°C-10°C. After the addition, the temperature was naturally raised to room temperature and reacted overnight. The reaction system was spin-dried, diluted with water (100 mL), extracted with EA (150 mL*3), the organic phases were combined, dried over _{anhydrous Na2SO4} , filtered, concentrated in vacuo, and purified by column chromatography (PE/EA=10/1, v/v) to obtain compound 16-1e (4.1 g), yield: 80.1%; MS: m/z 353.9 [M+1] ⁺ ;

The synthesis operation of compound 16-1f to compound 16-1 is basically the same as the synthesis operation of compound 1-2f to compound 1-2 in Comparative Example 1.

Compound 16-1f (yield 73.3%), MS: m/z 346.2 [M+1] ⁺ ; Compound 16-1g (yield 82.8%), MS: m/z 317.2 [M+1] ⁺ ; Compound 16-1h (yield 78.4%), MS: m/z 300.1 [M+1] ⁺ ; Compound 16-1 (yield 74.6%), MS: m/z 318.1 [M+1] ⁺ ;

(2) Synthesis route of compound 16

Under nitrogen protection, compound 16-1 (100.0 mg, 0.315 mmol, 1.0 eq), Tol (6 mL), compound 1-5 (101.0 mg, 0.63 mmol, 2.0 eq), Pd ₂ (dba) ₃ (173.3 mg, 0.19 mmol, 0.6 eq), Xantphos (218.9 mg, 0.38 mmol, 1.2 eq) and K ₃ PO ₄ (401.6 mg, 1.89 mmol, 6.0 eq) were added into a microwave tube in sequence, and the reaction was carried out at 110° C. for 2 h. The product was filtered, concentrated to remove the solvent, and purified by column chromatography (EA/MeOH=20/1, v/v) to obtain compound 16 (32.51 mg). The yield was 23.4%; MS: m/z 442.4[M+1] ⁺ .

Example 17: Preparation of Compound 17

Except that tetrahydrofuran-3(2H)-one was replaced by acetone and compound 1-1 was replaced by compound 1-5, other conditions were basically the same as those in Example 12 to prepare compound 17 (18.7 mg) with a yield of 24.7%; MS: m/z 386.3[M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ7.85-7.81 (m, 1H), 7.77 (d, J=8.1Hz, 1H), 7.71 (s, 1H), 7.64 (d, J=7.9Hz, 1H), 7.40 (s, 1H), 7.35 (t, J=7.9Hz, 1H), 5.44-5.38 (m, 1H), 4. 30-4.23 (m, 1H), 3.71 (t, J=8.4Hz, 2H), 3.20 (t, J=8.5Hz, 2H), 2.68 (s, 3H), 2.58 (d, J=7.8Hz, 3H), 1.59 (d, J=7.0Hz, 3H), 1.30-1.26 (m, 6H).

Compound 17-1a (yield 76.5%), MS: m/z 220.1 [M+1] ⁺ ; Compound 17-1b (yield 77.1%), MS: m/z 298.0 [M+1] ⁺ ; Compound 17-1c (yield 80.3%), MS: m/z 290.2 [M+1] ⁺ ; Compound 17-1d (yield 84.8%), MS: m/z 262.1 [M+1] ⁺ ; Compound 17-1e (yield 72.5%), MS: m/z 244.1 [M+1] ⁺ ; Compound 17-1f (yield 73.9%), MS: m/z 262.1 [M+1] ⁺ .

Example 18: Preparation of Compound 18

Except that tetrahydrofuran-3(2H)-one was replaced by cyclobutanone and compound 1-1 was replaced by compound 1-5, other conditions were basically the same as those in Example 12 to prepare compound 18 (15.3 mg) with a yield of 20.8%; MS: m/z 398.3[M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ8.05 (d, J=6.6Hz, 1H), 7.83 (d, J=7.9Hz, 1H), 7.71 (s, 1H), 7.63 (d, J=7.7, 1H), 7.47 (s, 1H), 7.35 (t, J=7.8Hz, 1H), 5.44-5.34 (m, 1H), 4.59 -4.53(m, 1H), 3.87-3.82(m, 2H), 3.21(t, J=8.3Hz, 2H), 2.68(s, 3H), 2. 61 (s, 3H), 2.39-2.23 (m, 4H), 1.79-1.70 (m, 2H), 1.61 (d, J=6.9Hz, 3H).

Compound 18-1a (yield 72.5%), MS: m/z 232.1 [M+1] ⁺ ; Compound 18-1b (yield 77.4%), MS: m/z 310.0 [M+1] ⁺ ; Compound 18-1c (yield 81.4%), MS: m/z 302.2 [M+1] ⁺ ; Compound 18-1d (yield 85.3%), MS: m/z 274.1 [M+1] ⁺ ; Compound 18-1e (yield 70.5%), MS: m/z 256.1 [M+1] ⁺ ; Compound 18-1f (yield 71.4%), MS: m/z 274.1 [M+1] ⁺ .

Example 19: Preparation of Compound 19

Except that tetrahydrofuran-3(2H)-one was replaced by cyclopentanone and compound 1-1 was replaced by compound 1-5, other conditions were basically the same as Example 12 to prepare compound 19 (25.3 mg) with a yield of 18.3%; MS: m/z 412.2 [M+1] ⁺ .

Compound 19-1a (yield 68.8%), MS: m/z 246.1 [M+1] ⁺ ; Compound 19-1b (yield 71.5%), MS: m/z 324.0 [M+1] ⁺ ; Compound 19-1c (yield 78.3%), MS: m/z 316.2 [M+1] ⁺ ; Compound 19-1d (yield 88.5%), MS: m/z 288.1 [M+1] ⁺ ; Compound 19-1e (yield 72.7%), MS: m/z 270.1 [M+1] ⁺ ; Compound 19-1f (yield 73.6%), MS: m/z 288.1 [M+1] ⁺ .

Example 20: Preparation of Compound 20

Except that tetrahydrofuran-3(2H)-one was replaced by tetrahydro-2H-pyran-3-one and compound 1-1 was replaced by compound 1-5, other conditions were basically the same as Example 12 to prepare compound 20 (15.3 mg) with a yield of 11.8%; MS: m/z 428.2 [M+1] ⁺ .

Compound 20-1a (yield 77.4%), MS: m/z 262.1 [M+1] ⁺ ; Compound 20-1b (yield 76.6%), MS: m/z 340.0 [M+1] ⁺ ; Compound 20-1c (yield 78.3%), MS: m/z 332.2 [M+1] ⁺ ; Compound 20-1d (yield 88.2%), MS: m/z 304.1 [M+1] ⁺ ; Compound 20-1e (yield 72.5%), MS: m/z 286.1 [M+1] ⁺ ; Compound 20-1f (yield 75.3%), MS: m/z 304.1 [M+1] ⁺ .

Example 21: Preparation of Compound 21

Except that tetrahydrofuran-3(2H)-one was replaced by tetrahydro-2H-pyran-4-one and compound 1-1 was replaced by compound 1-5, other conditions were basically the same as Example 12 to prepare compound 21 (9.3 mg) with a yield of 8.6%; MS: m/z 428.2 [M+1] ⁺ .

Compound 21-1a (yield 78.8%), MS: m/z 262.1 [M+1] ⁺ ; Compound 21-1b (yield 75.8%), MS: m/z 340.0 [M+1] ⁺ ; Compound 21-1c (yield 79.3%), MS: m/z 332.2 [M+1] ⁺ ; Compound 21-1d (yield 87.4%), MS: m/z 304.1 [M+1] ⁺ ; Compound 21-1e (yield 75.3%), MS: m/z 286.1 [M+1] ⁺ ; Compound 21-1f (yield 73.9%), MS: m/z 304.1 [M+1] ⁺ .

Example 22: Preparation of Compound 22

Except that tetrahydrofuran-3(2H)-one was replaced by N-Cbz-pyrrolidin-3-one, other conditions were basically the same as those in Example 12 to obtain compound 22a (45 mg), MS: m/z 547.3 [M+1] ⁺ ;

Compound 22a (45 mg), palladium carbon (20 mg) and methanol (5 mL) were added to a reaction bottle, and the hydrogen was replaced three times. The reaction was carried out at 55°C for 48 h. The reaction was completed by mass spectrometry monitoring and purification (dichloromethane/methanol: 5/1, v/v) to obtain compound 22 (5.3 mg). MS: m/z 427.2 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ7.94 (s, 1H), 7.80 (d, J = 7.9Hz, 1H), 7.74 (s, 1H), 7.64 (d, J = 7.6Hz, 1H), 7.55 (s, 1H), 7.35 (t, J=7.8Hz, 1H), 5.44-5.38 (m, 1H), 4.82-4.77 (m, 1H), 3.99 (q, J=7.7Hz, 1 H), 3.94-3.84 (m, 2H), 3.78 (q, J=8.5Hz, 3H), 3.57 (s, 3H), 3.21 (t, J=8.3Hz, 2H), 2. 69 (s, 3H), 2.61 (s, 3H), 2.40-2.30 (m, 1H), 2.09-2.04 (m, 1H), 1.61 (d, J=6.9Hz, 3H).

Compound 22-1a (yield 69.7%), MS: m/z 381.2 [M+1] ⁺ ; Compound 22-1b (yield 67.4%), MS: m/z 459.1 [M+1] ⁺ ; Compound 22-1c (yield 71.5%), MS: m/z 451.2 [M+1] ⁺ ; Compound 22-1d (yield 84.4%), MS: m/z 423.2 [M+1] ⁺ ; Compound 22-1e (yield 79.5%), MS: m/z 405.2 [M+1] ⁺ ; Compound 22-1f (yield 70.7%), MS: m/z 423.1 [M+1] ⁺ .

Example 23: Preparation of Compound 23

Except that 1-1 was replaced by 1-3, other conditions were basically the same as those in Example 12 to obtain compound 23a (18.0 mg) with a yield of 26.5%; MS: m/z 600.3 [M+1] ⁺ ;

Compound 23a (18 mg) was dissolved in dichloromethane (1.5 mL), trifluoroacetic acid (0.5 mL) was added, and the mixture was reacted at room temperature for 2 h. After the reaction was completed, saturated sodium bicarbonate solution was added to adjust the pH to 7-8, and the mixture was extracted with dichloromethane (10 mL*3), concentrated, and purified by thin layer chromatography (dichloromethane/methanol=5/1, v/v) to obtain compound 23 (13 mg) with a yield of 87%; MS: m/z 500.2[M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ): δ7.94 (s, 1H), 7.82-7.80 (m, 2H), 7.74 (s, 1H), 7.66-7.63 (m, 2H), 7.55 (s, 1H), 7 .32(s, 1H), 7.25(s, 1H), 5.44-5.38(m, 1H), 4.82-4.77(m, 1H), 4.25(s, 1H), 3.99(q , J=7.7Hz, 1H), 3.94-3.84 (m, 4H), 3.78 (q, J=8.5Hz, 3H), 3.46 (s, 3H), 3.21 (t, J=8. 3Hz, 2H), 2.69 (s, 3H), 2.40-2.30 (m, 1H), 2.09-2.04 (m, 1H), 1.61 (d, J=6.9Hz, 3H).

Experimental Example 1: KRAS-G12C/SOS1 IC ₅₀ Activity Test

The inhibition rate of KRAS-G12C/SOS1 was detected by using the BindingAssay method to evaluate the inhibitory effect of the compound of the present invention on KRAS-G12C/SOS1.

1.1 Experimental Materials

1.1.1 Reagents and consumables

1.1.2 Instruments

Centrifuge (Manufacturer: Eppendorf, Model: 5430)

Microplate reader (Manufacturer: Perkin Elmer, Model: Envision)

Echo 550 (Manufacturer: Labcyte, Model: Echo 550)

1.2 Kinase reaction process

(1) Compound preparation: The concentration gradient of the test compound is 500 nm, 125 nm, 31.25 nm, 7.81 nm, 1.95 nm, 0.488 nm, 0.122 nm and 0.03 nm. The test is performed in duplicate wells and diluted to 200 times the final concentration of 100% DMSO solution in a 384-well plate. Use a dispenser Echo550 to transfer 50 nl of the 200 times final concentration of the compound to the target 384-well plate. Add 50 nl of 100% DMSO to the negative control well and the positive control well, respectively.

(2) Prepare a Tag1-SOS1 solution with a 4-fold final concentration using dilution buffer.

(3) Add 2.5 μl of Tag1-SOS1 solution at 4 times the final concentration to a 384-well plate.

(4) Prepare a Tag2-KRAS-G12C solution with a 4-fold final concentration using dilution buffer.

(5) Add 2.5 μl of Tag2-KRAS-G12C solution at 4 times the final concentration to the compound wells; add 2.5 μl of dilution buffer to the negative control wells.

(6) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, shake to mix, and incubate at room temperature for 15 minutes.

(7) Prepare 1x final concentration of Anti-Tag1-TB3+ solution and 1x final concentration of Anti-Tag2-XL665 solution using detection buffer. After mixing the two solutions, add 5 μl of Mix solution to each well.

(8) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, shake to mix, and incubate at room temperature for 120 minutes.

(9) Read the results using Envision microplate reader and record Em665/620.

1.3 Data Analysis

1.3.1 Inhibition rate calculation formula

Among them: Min signal represents the average absorbance of pure DMSO wells; Max signal represents the average absorbance of enzyme, substrate, detection reagent, and DMSO mixture wells; Compound signal represents the average absorbance of sample wells.

1.3.2 Fitting the dose-effect curve

The log value of the compound concentration was used as the X-axis and the percentage inhibition rate was used as the Y-axis. The log (inhibitor) vs. response-Variable slope of the analysis software GraphHPad Prism 5 was used to fit the dose-effect curve to obtain the IC ₅₀ value of each compound on the enzyme activity.

The fitting formula is: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC ₅₀ -X) × HillSlope))

Among them: Top represents the top platform, and the Top standard of the curve is generally between 80% and 120%; Bottom represents the bottom platform, and the Bottom of the curve is generally between -20% and 20%.

The results of in vitro inhibitory activity test (expressed as IC ₅₀ ) are shown in Table 1.

Table 1. Effects of the compounds of the present invention on KRAS-G12C/SOS1

As can be seen from Table 1, the compounds of the present invention have strong inhibitory activity against KRAS-G12C/SOS1, and are significantly superior to positive control compound 2 and positive control compound 3 (positive control compound 3 is derived from compound 107 of patent CN115536660A).

Experimental Example 2: Effects of compounds on ERK phosphorylation of SOS1 downstream protein in NCI-H358 and MKN1 cells

1. Experimental materials, consumables and instruments

1.1 Materials

1.2 Consumables

1.3 Instruments

2. Experimental Methods

The final concentrations of the test compounds were 10 μM, 3.33 μM, 1.11 μM, 0.37 μM, 0.123 μM, 0.041 μM, 0.0137 μM, 0.0045 μM, 0.0015 μM and 0.00051 μM. After the gradient dilution compounds were incubated with the cells overnight for 3 hours, the cells were fixed, permeated and blocked, and incubated overnight in the primary antibody. After that, the secondary antibody labeled with infrared fluorescent dye was used to detect the fluorescence signal using the Odyssey two-color infrared fluorescence imaging system.

The specific steps are as follows:

Day 1: NCI-H358/MKN1 cells were seeded in a 384-well cell culture plate and cultured overnight in a 37°C, 5% carbon dioxide cell culture incubator.

The next day: Add the compound to the plate using Echo550 and continue to culture in a 37°C, 5% CO2 cell culture incubator for 3 hours. Finally, the cell culture plate was fixed with paraformaldehyde, permeabilized with methanol, and blocked with blocking solution, and then the primary antibody mixture (rabbit anti pERK, mouse anti GAPDH) was added and incubated at 4°C overnight.

Day 3: Discard the primary antibody and add the secondary antibody mixture (goat anti rabbit 800CW, goat anti mouse 680RD), incubate at room temperature in the dark, wash with PBST, centrifuge the cell culture plate upside down for 1 minute, and read the fluorescence signal value with Odyssey CLx.

3. Data Analysis

ERK phosphorylation was calculated according to the following formula:

Relative inhibition rate (%) = (fluorescence signal ratio of compound - average value of positive control) / (average value of negative control - average value of positive control) * 100%;

Wherein: the negative control is DMSO, and the positive control is the positive control compound 1 mentioned above.

XLFit 5.0 calculates _IC50 values using the parameter fitting formula:

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)×HillSlope));

Wherein: X is the log value of compound concentration, Y is the average relative inhibition rate of p-ERK between duplicate wells, Bottom is the minimum inhibition rate of the compound, and Top is the maximum inhibition rate of the compound.

4. Experimental Results

The IC ₅₀ results of the compounds of the present invention are shown in Table 2.

Table 2. Effects of the compounds of the present invention on phosphorylation of ERK, a downstream protein of SOS1 in NCI-H358 and MKN1 cells

As shown in Table 2, the compounds of the present invention can significantly inhibit the phosphorylation of pERK downstream of NCI-H358 and MKN1, and have excellent anti-cell proliferation activity.

Experimental Example 3: Effects of Compounds on Cytochrome P450 Enzymes CYP3A4, CYP2B6 and CYP1A2

1. Experimental Materials

Human liver microsomes, catalog number 452117, purchased from Corning;

NADPH, product number BD11658, was purchased from Bidler.

2. Experimental Methods

(1) Add 1 μL of 2 mM test compound or quality control inhibitor (ketoconazole for CYP3A4 and CYP2B6 inhibitors, furafylline for CYP1A2 inhibitor), and the final concentration of the test compound or quality control inhibitor is 10 μM; DMSO is used as a negative control, and all experiments are performed in duplicate;

(2) Prepare a mixed solution system containing phosphate, ultrapure water, _MgCl2 and human liver microsomes according to the following table;

(3) Take 1 μL of the substrate stock solution prepared as shown in the table below and add it to the above mixed solution;

(4) Preheat the reaction system in a 37°C water bath for 5 min, then add 20 μL of 10 mM NADPH solution to start the reaction, with a final NADPH concentration of 1 mM, and incubate at 37°C for the time shown in the table above;

(5) After the incubation, at the specified time point, 400 μL of cold methanol (internal standard, 500 nM Labetalol, 100 nM Alprazolam and 2 μM Ketoprofen) was added to terminate the reaction. The sample was centrifuged at 3220 g for 60 min to precipitate the protein. 100 μL of the supernatant was taken and diluted with 100 μL ultrapure water (according to the peak shape and signal intensity of LC-MS/MS) for LC-MS/MS analysis.

3. Experimental results

The IC ₅₀ results of the compounds of the present invention are shown in Table 3.

Table 3. Results of CYP enzyme inhibition rate of the compounds of the present invention

As shown in Table 3, under the condition of 10 μM, the compounds of the present invention did not show strong inhibition on each CYP enzyme subtype, and the DDI risk was low.

Experimental Example 4: Plasma kinetic study of compounds in CD-1 mice

1. Experimental Animals

CD-1 mice, SPF grade, male, were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd.

2. Experimental Methods

The compounds were tested for pharmacokinetic properties in rodents following oral and intravenous administration using standard protocols.

In the experiment, the candidate compound was prepared into a clear solution and given to mice orally and intravenously in a single dose, with the solvent being pH 4.5 acetate buffer. Six mice were used for oral and intravenous administration of each compound, with an oral dose of 10 mg/kg (drug solution concentration of 1 mg/mL and administration volume of 10 μL/g); the intravenous dose was 3 mg/kg (drug solution concentration of 0.3 mg/mL and administration volume of 10 μL/g).

After administration, samples were collected at 0.0833h, 0.25h, 0.5h, 1h, 2h, 4h, 8h and 24h. Whole blood samples were immediately placed on ice and centrifuged at 4°C, 2000g for 5min within 0.5h to separate plasma. The upper layer samples were collected into sample tubes and then lysed within 0.5h. Freeze in a -10 to -30°C refrigerator and transfer to a -60 to -90°C refrigerator within 24 hours.

3. Data Analysis

The concentration of each prototype drug in plasma samples at each time point was detected by LC-MS/MS method, and PK parameters such as plasma clearance (CL), peak time (T _max ), half-life (t _1/2 ), peak concentration (C _max ), area under the concentration-time curve (AUC _0-t ), tissue distribution volume (vd), mean residence time (MRT _0-t ) and bioavailability (F) were calculated using WinNonlin software.

4. Experimental Results

The pharmacokinetic test results of the compounds of the present invention are shown in Table 4. Table 4. Pharmacokinetic test results of the compounds of the present invention

As shown in Table 4, the compounds of the present invention have the characteristics of low plasma clearance, long drug half-life, good exposure and high bioavailability, and have good pharmacokinetic properties. The results of compounds 1, 13 and 14 are better than those of the positive control compound 1.

Experimental Example 5: In vivo pharmacodynamic study of the compound in the human lung cancer NCI-H358 subcutaneous xenograft tumor model

1. Experimental Animals

NCG mice, SPF grade, female, were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.

2. Experimental Methods

NCI-H358 tumor cells (5×10 ⁷ /mL, 100 μL/mouse) in the logarithmic growth phase were inoculated into 6-8 week old female NCG nude mice (weighing about 20 g) and all mice were subcutaneously inoculated. The mice were cultured in an SPF-grade experimental environment and all mice had free access to commercially certified standard diets.

When the average tumor volume of mice grew to about 150 ^mm3 , they were divided into groups, with 6 mice in each group, and the test compound was orally administered daily (twice a day, 50 mpk each time, the solvent was 5% DMSO, 5% Tween 80, 40% PEG400, 50% water); blank group (5% DMSO, 5% Tween 80, 40% PEG400, 50% water).

The tumor volume was measured every two days with a two-dimensional caliper, and the animals were weighed every day. After 21 consecutive days of administration, the inhibition rate (TGI/100%) was calculated based on the final tumor volume.

The tumor volume was calculated using the following formula:

V = 1/2*a*b ² ;

Where: a represents the long diameter of the tumor, and b represents the short diameter of the tumor.

Table 5. In vivo pharmacodynamics test results of the compounds of the present invention

As shown in FIG1 , FIG2 and Table 5 , the compounds of the present invention have good in vivo efficacy and are superior to the positive control compound 1.

Although the present invention has been described by way of specific examples above, it should not be construed as being limited thereto, but rather the invention encompasses the general aspects previously disclosed and is susceptible to numerous modifications and embodiments without departing from the spirit and scope of the invention.

Experimental Example 6: In vivo pharmacodynamic study of the compound of the present invention in the human pancreatic cancer NCI-H358 subcutaneous xenograft tumor model

1. Experimental Animals

NCG mice, SPF grade, female, were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.

2. Experimental Methods

NCI-H358 tumor cells (5×10 ⁷ /mL, 100 μL/mouse) in the logarithmic growth phase were inoculated into 6-8 week old female NCG nude mice (weighing about 20 g) and all mice were subcutaneously inoculated. The mice were cultured in an SPF-grade experimental environment and all mice had free access to commercially certified standard diets.

When the average tumor volume of mice grew to about 130 mm ³ , they were divided into groups, 6 mice in each group, and the test compound was orally administered daily (the drug solvent was 5% DMSO, 5% Tween 80, 40% PEG400, 50% water).

The tumor volume was measured every two days with a two-dimensional caliper, and the animals were weighed every day. After 21 consecutive days of administration, the inhibition rate (TGI/100%) was calculated based on the final tumor volume.

The tumor volume was calculated as follows:

V = 1/2 × a × b ² ;

Where: a represents the long diameter of the tumor, and b represents the short diameter of the tumor.

Table 6. In vivo pharmacodynamics test results of the compounds of the present invention

As shown in Table 6, the combined administration of compound 14 of the present invention and trametinib or cediranib has better in vivo efficacy, and the tumor inhibition rate is more than 90%.

Experimental Example 7: In vivo pharmacodynamic study of the compound of the present invention in a human pancreatic cancer Miapaca2 subcutaneous xenograft tumor model

1. Experimental Animals

NCG mice, SPF grade, female, were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.

2. Experimental Methods

Miapaca2 tumor cells (5×10 ⁷ /mL, 100 μL/mouse) in the logarithmic growth phase were inoculated into 6-8 week old female NCG nude mice (weighing about 20 g) and all mice were subcutaneously inoculated. The mice were cultured in an SPF-grade experimental environment and all mice had free access to commercially certified standard diets.

When the average tumor volume of mice grew to about 130 ^mm3 , they were divided into groups, with 6 mice in each group, and the test compound was orally administered daily (twice a day, compound 14, 50 mpk each time, the solvent was 5% DMSO, 5% Tween 80, 40% PEG400, 50% water; trametinib, 0.125 mpk each time, the solvent was 5% DMSO, 5% Tween 80, 40% PEG400, 50% water); blank group (5% DMSO, 5% Tween 80, 40% PEG400, 50% water).

The tumor volume was measured every two days with a two-dimensional caliper and the animals were weighed every day. After 26 consecutive days of administration, the inhibition rate (TGI/100%) was calculated based on the final tumor volume.

The tumor volume was calculated as follows:

V = 1/2 × a × b ² ;

Where: a represents the long diameter of the tumor, and b represents the short diameter of the tumor.

Table 7. In vivo pharmacodynamics test results of the compounds of the present invention

As shown in Table 6, the combined administration of compound 14 of the present invention and trametinib has better in vivo efficacy, and the tumor inhibition rate reaches 90.1%.

Experimental Example 8: Toxicity test of SD rats given the compound of the present invention by oral gavage for repeated administration for 2 weeks

21 SD rats were randomly divided into 3 dose groups according to body weight, including vehicle control group, low-dose group and high-dose group of compound 14 (dosage was 0, 200, 300 mg/kg/day, respectively), toxicity test group and toxicity satellite group. There were 2 rats/group/sex in the toxicity test group; there were 3 male rats/group in the toxicity satellite group. Each dose group of compound 14 was given the corresponding concentration of compound 14 dosage preparation by oral gavage at a dosage volume of 10 mL/kg, and the vehicle control group was given 5% DMSO + 5% Tween 80 + 40% PEG400 + 50% pH 4.5 acetate buffer at the same dosage volume. The drug was administered twice a day for 14 consecutive days (28 times in total), and the day of the first administration was defined as the first day of the experiment (D1).

During the experiment, the general condition of the animals in each group was observed every day, and the body weight was measured twice a week; the food intake of the animals in the toxicity test group was measured once a week; at the end of administration (D15), the hematology, coagulation, and blood biochemistry of the animals in the toxicity test group were tested, and 2 animals/group/sex were dissected, and gross anatomical observation, organ weighing, and histopathological examination were performed after euthanasia.

Blood samples were collected from animals in the toxic satellite group compound 14 group before the first and last administration and 0.5h, 1h, 2h, 4h, 4.5h, 5h, 8h, and 24h after administration, and blood samples were collected from animals in the toxic satellite group vehicle control group before the first and last administration and 1h after administration to detect the concentration of compound 14 in plasma. Phoenix WinNonlin was used to calculate toxicokinetic parameters such as C _max , T _max , and AUC _last .

Under the conditions of this experiment, SD rats were given 200 mg/kg/day and 300 mg/kg/day of compound 14 by oral gavage twice a day for 2 consecutive weeks (a total of 28 times), and no animals in each group were dying/dead or had other serious toxic reactions. During the experiment, no obvious abnormal changes with toxicological significance related to compound 14 were found in the general condition, body weight, food intake, hematology, blood coagulation, blood biochemistry, organ weight and coefficient of the animals at each test point.

Toxicokinetic results showed that after the first and last administration, the AUC _last and C _max of compound 14 in SD rats increased with the increase of administration dose (at the first and last administration of 200 mg/kg/day, the AUC of male rats were 62996 h·ng/mL and 85232 h·ng/mL, respectively; at the first and last administration of 300 mg/kg/day, the AUC of male rats were 112966 h·ng/mL and 121246 h·ng/mL, respectively); the ratios of AUClast (last/first) were 1.35 and 1.07, respectively, indicating that the compound had no obvious accumulation trend after 14 days of continuous administration.

The tissue distribution results showed that at the end of the dosing period, the concentrations of compound 14 in the liver, kidney, pancreas, lung, heart, spleen and stomach of SD rats increased with increasing doses, and the distribution order was: stomach > liver > kidney > lung > spleen ≈ pancreas > heart.

Gross autopsy showed that there were no obvious test substance-related histopathological changes in the heart, liver, spleen, lung, kidney, stomach, and pancreas of the animals in the high-dose group of the toxicity test group of this experiment.

In summary, SD rats were given 200 mg/kg/day and 300 mg/kg/day of compound 14 by oral gavage twice a day for 2 consecutive weeks (28 times in total), and the maximum tolerated dose (MTD) was > 300 mg/kg/day.

Claims (17)

A compound of formula I or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof,
in: Every independently represent a single bond or a double bond; X is selected from CR ₆ , C(R ₆ ) ₂ and C(═O); Y is selected from CR ₇ and C(R ₇ ) ₂ ; Z is selected from N and NR ⁸ , or Z is absent; when Z is absent, Y is directly connected to the phthalazine ring via a single bond; Ring A is selected from phenyl and thienyl (especially thien-2-yl); each R ¹ is independently selected from methyl, ethyl, phenyl, halogen and cyano, wherein the methyl, ethyl or phenyl is optionally substituted with one or more substituents selected from methylaminomethyl, dimethylaminomethyl, hydroxyl and halogen; ^R2 and ^R3 are independently hydrogen; ^R4 is methyl; R ⁵ is selected from C _1-8 alkyl, C _3-6 cycloalkyl and 3-8 membered heterocyclyl, wherein the C _1-8 alkyl is optionally substituted by one or more deuterium; the 3-8 membered heterocyclyl contains a heteroatom selected from O, N and S, and the 3-8 membered heterocyclyl is optionally substituted by one or more C _1-8 alkyl; If present, each R ⁶ is independently hydrogen; Each R ⁷ is independently selected from hydrogen and methyl; or, two R ⁷ together with the carbon atom to which they are attached form a cyclopropyl group; If present, R ⁸ is selected from methyl and cyclopropyl, wherein the methyl is optionally substituted with one or more deuterium; n is 1 or 2; Furthermore, when Z is absent, Y is C(R ₇ ) ₂ , and X is C(═O), the two R ⁷ are not connected to each other and are different from each other; Also, does not contain the following compounds:
The compound according to claim 1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: Ring A is selected from phenyl and thiophen-2-yl. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, characterized in that: Each R ¹ is independently selected from -CH ₃ , -CH ₂ F, -CHF ₂ , -CF ₃ , -CH ₂ OH, -CH ₂ CH ₃ , -CHFCH ₃ , -CF ₂ CH ₃ , -CH ₂ CH ₂ OH, -CHFCH ₂ OH, -CF ₂ CH ₂ OH, -F, -Cl, -Br, -CN, Preferably, each R ¹ is independently selected from -CH ₃ , -CHF ₂ , -CF ₂ CH ₂ OH, -F, -CN, The compound according to claim 1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: The compound is shown in Formula IA or Formula IA':
in: Every independently represent a single bond or a double bond; R ¹ is selected from X, Y, Z and R ⁵ are as defined in claim 1. The compound according to claim 1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: The compound is shown in Formula IB or Formula IB':
in: Every independently represent a single bond or a double bond; each R ¹ is independently selected from -CH ₃ , -CHF ₂ , -CF ₂ CH ₂ OH, -F and -CN; X, Y, Z and R ⁵ are as defined in claim 1. The compound according to claim 5 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: Selected from Best The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof, characterized in that: Selected from Preferably, Selected from The compound according to claim 1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: R ⁵ is selected from C _1-4 alkyl, C _3-6 cycloalkyl and 3-6 membered heterocyclyl, wherein the C _1-4 alkyl is optionally substituted by one or more deuterium; the 3-6 membered heterocyclyl contains a heteroatom selected from O and N, and the 3-6 membered heterocyclyl is optionally substituted by one or more C _1-4 alkyl; Preferably, R ⁵ is selected from -CH ₃ , -CH ₂ D, -CHD ₂ , -CD ₃ , isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, pyrrolidin-3-yl, 1-methylpyrrolidin-3-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl and tetrahydrofuran-3-yl; More preferably, ^R5 is selected from -CH3, _-CD3 , _isopropyl , cyclopropyl, cyclobutyl, cyclopentyl, 1-methylpyrrolidin-3-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl and tetrahydrofuran-3-yl. The compound according to claim 1 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: R ⁸ is selected from -CH ₃ , -CH ₂ D, -CHD ₂ , -CD ₃ and cyclopropyl; Preferably, R ⁸ is selected from -CH ₃ , -CD ₃ and cyclopropyl. The compound according to claim 4 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: The compound is shown in Formula IA-1 or Formula IA'-1:
Wherein: R ¹ is as defined in claim 4. The compound according to claim 4 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: The compound is shown in Formula IA-2 or Formula IA'-2:
Wherein: R ¹ is as defined in claim 4. The compound according to claim 5 or its pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug, characterized in that: The compound is as shown in any one of formulas IB-1 to IB-16 or any one of formulas IB'-1 to IB'-16, preferably any one of formulas IB-1, IB-2 and IB-5 or any one of formulas IB'-1, IB'-2 and IB'-5:

Wherein: R ¹ is as defined in claim 5. A compound or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, An isotope-labeled substance or prodrug, characterized in that The compound is selected from the following compounds:

A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled substance or prodrug thereof; Preferably, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier. A pharmaceutical preparation, which is prepared from the pharmaceutical composition according to claim 14; Preferably, the pharmaceutical preparation is a solid preparation for oral administration, a liquid dosage form for oral administration, a dosage form for parenteral injection or a dosage form for topical administration. Use of a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotope-labeled or prodrug thereof, a pharmaceutical composition according to claim 14, or a pharmaceutical preparation according to claim 15 in the preparation of a medicament for preventing and/or treating a disease mediated at least in part by SOS1 protein; Preferably, the disease at least partly mediated by SOS1 protein is cancer, such as pancreatic cancer, non-small cell lung cancer and colorectal cancer. A pharmaceutical composition comprising a compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt, hydrate, solvate, stereoisomer, tautomer, cis-trans isomer, isotopically labeled or prodrug thereof, a pharmaceutical composition according to claim 14, or a pharmaceutical preparation according to claim 15, and at least one additional cancer therapeutic agent; Preferably, the cancer therapeutic agent is selected from trametinib and cediranib.