Compounds as wrn inhibitors and uses thereof

Abstract

The present application belongs to the field of medicinal chemistry, and relates to a kind of as WRN inhibitor Compound and its application, specifically, the present application provides the compound shown in formula (I) and formula (II) or its pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug, their preparation method and the pharmaceutical composition containing these compounds and the purposes of these compounds or compositions for treating WRN related diseases.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry, specifically relating to compounds as WRN inhibitors or their pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs, methods for their preparation, pharmaceutical compositions containing these compounds, and the use of these compounds or compositions for the treatment of WRN-related diseases. Background Technology

[0002] Microsatellites are short tandemly repetitive DNA sequences in the genome, typically composed of 1-6 nucleotides. Due to variations in the number of repetitions of their core repeat units, microsatellites often exhibit population polymorphism. The phenomenon where new microsatellite alleles appear at a microsatellite locus in a tumor due to the insertion or deletion of repeat units, compared to normal tissue, is called microsatellite instability (MSI). MSI occurs because of functional defects in DNA mismatch repair (dMMR) in tumor tissue. With the advancement of preclinical and clinical research, MSI accompanied by DNA mismatch repair defects has become an important tumor marker in clinical practice. Clinically, dMMR and MSI-H usually have the same clinical significance. Cancers that develop MSI include colorectal cancer, gastric cancer, endometrial cancer, ovarian cancer, bile duct cancer, urethral cancer, brain cancer, and skin cancer. Among these, colorectal cancer is the most common (MSI-H ≈ 15%). In multiple clinical trials, PD-1 monoclonal antibodies, represented by pembrolizumab, have been found to have good therapeutic effects on MSI-H tumors. However, a significant proportion of MSI patients in these studies do not respond to immunotherapy, and some MSI-H tumor patients develop primary resistance to immunotherapy. The clinical needs of these patients remain unmet.

[0003] Recent studies have revealed a strong correlation between WRN (Werner Protein) in the RecQ family of RNA / DNA helicases and the development of MSI-H tumors. Knocking out or downregulating WRN in MSI-H tumor cells significantly inhibits tumor cell proliferation, invasion, and migration, and promotes tumor cell apoptosis. These findings suggest a possible synthetic lethal relationship between WRN and MSI-H tumors. The mechanism by which WRN knockdown promotes apoptosis in MSI-H tumors has been elucidated: TA-dinucleotide repeats are highly unstable in MSI cells and undergo massive amplification. These increased TA-dinucleotide repeats form non-B DNA secondary structures, halting replication forks and activating ATR checkpoint kinases. This process requires the participation of WRN helicase to reactivate DNA replication forks and allow DNA replication to continue. Without WRN, the amplified TA-dinucleotide repeat sequences are easily cleaved by MUS81 nuclease, leading to extensive chromosome fragmentation and inducing apoptosis.

[0004] Therefore, the development of WRN small molecule inhibitors is expected to provide new means of anti-tumor treatment, enabling MSI-H tumor patients to obtain clinical benefits. Summary of the Invention

[0005] This invention provides compounds of general formula (I) or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof.

[0006]

[0007] in,

[0008] X is selected from CH and N;

[0009] Q is selected from =O, =NH, and =NCH3;

[0010] W is selected from halogen, cyano, amino, alkyl, cycloalkyl, heterocyclic, haloalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, alkylamino, aryl, heteroaryl, arylalkyl, heteroarylalkyl, cycloalkylalkyl, and heterocyclicalkyl, optionally substituted by one or more groups selected from halogen, hydroxy, amino, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, aryl, heteroaryl, cycloalkyl, and heterocyclicalkyl;

[0011] R 1 Selected from hydrogen, halogen, cyano, amino, alkyl, oxo group, cycloalkyl, heterocyclic, heterocyclic alkenyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, monoalkylamino, dialkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, aryl, and heteroaryl; and

[0012] R 2 It is selected from alkyl, haloalkyl, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclic, aryl and heteroaryl groups, and is optionally substituted by one or more groups selected from halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, amino, monoalkylamino, alkylacylamino, alkylacyl, alkylsulfonyl, aminoacyl, alkylaminoacyl, dialkylamino, haloalkylacyl, hydroxyalkylacyl, cycloalkyl, heterocyclic, aryl and heteroaryl groups.

[0013] In some embodiments, the compounds of the present invention are compounds of general formula (I) or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof, wherein general formula (I) has the structure of the following general formula (Ia):

[0014]

[0015] Among them, X, Q, W, and R 1 R 2 As defined in general formula (I).

[0016] In some specific embodiments, the compounds of the present invention are compounds of general formula (I) or (Ia) or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof, wherein W is selected from halogens, cyano groups, amino groups, C... 1-6 Alkyl, C 3-10 cycloalkyl, C 3-10 Heterocyclic groups, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, C 1-6 Alkylamino, C 6-10 Aryl, 5-10 heteroaryl, C 6-10 Aryl C 1-6 Alkyl, 5-10-membered heteroaryl C 1-6 Alkyl, C 3-10 cycloalkyl C 1-6 Alkyl and C 3-10 Heterocyclic C 1-6 Alkyl groups, optionally composed of one or more elements selected from halogens, hydroxyl groups, amino groups, C4 groups, and hydroxyl groups. 1-6 Alkyl, Halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, C 6-10 Aryl, 5-10 heteroaryl, C 3-10 cycloalkyl and C 3-10The heterocyclic alkyl group is substituted, wherein the heteroatom is selected from N, O, S, and P. In some specific embodiments, W is selected from methyl, ethyl, ... , , , , and .

[0017] In some embodiments, a compound of formula (I) or (Ia) according to the invention, or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, wherein R 1 Selected from hydrogen, halogen, cyano, amino, C 1-6 Alkyl, oxo group, C 3-10 Cycloalkyl, 3-10 heterocyclic, 3-10 heterocyclic alkenyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, single C 1-6 Alkylamino, bis(C) 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, bis(C) 1-6 Alkylamino, C 6-10 Aryl and 5-10 heteroaryl groups, wherein the heteroatom is selected from N, O, S, and P. In some specific embodiments, R 1 It does not exist. In some other specific implementations, R 1 Selected from fluorine, chlorine, bromine, methyl, ethyl, propyl, oxo group, cyclopropyl, cyclobutyl, difluoromethyl, trifluoromethyl, and amino.

[0018] In some embodiments, a compound of formula (I) or (Ia) according to the invention, or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, wherein R 2 Selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, C 3-10 Cycloalkyl, 3-10 membered heterocyclic groups, C 6-10 aryl and 5-10 heteroaryl groups, optionally separated by one or more elements selected from halogen, hydroxyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6Alkoxy, amino, C 1-6 Monoalkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, bis(C) 1-6 Alkylamino, halogenated C 1-6 Alkyl acyl, hydroxy C 1-6 Alkyl acyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic groups, C 6-12 The aryl group and the 5-12-membered heteroaryl group are substituted, wherein the heteroatom is selected from N, O, S and P;

[0019] More preferably, R 2 Selected from C 1-3 Alkyl, Halogenated C 1-4 Alkyl, Halogenated C 1-3 Alkoxy, hydroxy C 1-3 Alkoxy, C 3-6 Cycloalkyl, 3-10 membered heterocyclic groups, C 6-10 aryl and 5-10 heteroaryl groups, optionally separated by one or more elements selected from halogen, hydroxyl, C 1-3 Alkyl, Halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxy C 1-3 Alkoxy, amino, C 1-3 Monoalkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, C 1-3 alkylsulfonyl, aminoacyl, C 1-3 Alkylaminoacyl, bis(C) 1-3 Alkylamino, halogenated C 1-3 Alkyl acyl, hydroxy C 1-3 Alkyl acyl, C 3-6 cycloalkyl, 3-6 membered heterocyclic, C 6-12 The aryl group and the 5-8-membered heteroaryl group are substituted, wherein the heteroatom is selected from N, O, S and P.

[0020] In some specific embodiments, a compound of formula (I) or (Ia) according to the invention, or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, wherein R 2 Selected from methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, phenyl, difluoromethyl, trifluoromethyl, , , and .

[0021] This invention provides compounds of general formula (II) or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof.

[0022]

[0023] Where Y is selected from O and NH, and n is selected from 0, 1, 2, 3 and 4.

[0024] In some embodiments, the compounds of the present invention are compounds of general formula (II) or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof, wherein general formula (II) has the structure of general formula (IIa):

[0025]

[0026] Where Y and n are as defined in general formula (II).

[0027] This invention provides the following specific compounds or their pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs:

[0028] , , , , , , , , , , , ,

[0029] ,and .

[0030] On the other hand, the present invention provides pharmaceutical compositions comprising the compounds of the present invention or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof.

[0031] In some embodiments, the present invention provides compounds of the present invention or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof, and pharmaceutical compositions comprising compounds of the present invention or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof, said compounds or pharmaceutical compositions for treating diseases associated with WRN.

[0032] In some embodiments, the present invention provides pharmaceutical compositions comprising the compound of the present invention or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug and a pharmaceutically acceptable carrier.

[0033] The compounds of the present invention, or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof, can be mixed with pharmaceutically acceptable carriers, diluents, or excipients to prepare pharmaceutical formulations suitable for oral or parenteral administration. Administration methods include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, and oral routes. The formulations can be administered via any route, such as by infusion or bolus, or by absorption through the epithelium or mucous membranes (e.g., oral mucosa or rectum). Administration can be systemic or local. Examples of oral formulations include solid or liquid dosage forms, specifically including tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The formulations can be prepared by methods known in the art and contain carriers, diluents, or excipients conventionally used in the field of pharmaceutical formulations.

[0034] Thirdly, the present invention provides a method for treating diseases associated with WRN using compounds of formulas (I) and (II) of the present invention or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof, or pharmaceutical compositions comprising such compounds, and their use in the preparation of medicaments for treating diseases associated with WRN.

[0035] In some preferred embodiments, the present invention provides a method for treating WRN-related diseases using compounds of formulas (I) and (II) of the present invention, or pharmaceutically acceptable salts, isomers, solvates, crystals, or prodrugs thereof, or pharmaceutical compositions comprising thereof, and their use in the preparation of medicaments for treating WRN-related diseases, wherein the WRN-related diseases include, but are not limited to, cancer, proliferative disorders, hematologic disorders, or metabolic disorders. In some embodiments, the WRN-related disease of the present invention is cancer.

[0036] In some embodiments, the WRN-related diseases described in this invention 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.

[0037] In some embodiments, the present invention provides the use of compounds of formulas (I) and (II) of the present invention, or pharmaceutically acceptable salts, isomers, solvates, crystals or prodrugs thereof, or pharmaceutical compositions comprising thereof, in the preparation of medicaments for treating and / or preventing microsatellite instability (MSI) or DNA mismatch repair (dMMR) cancers.

[0038] In some embodiments, the microsatellite instability (MSI) or DNA mismatch repair (dMMR) cancers described in this invention include, but are not limited to: colorectal cancer, gastric cancer, prostate cancer, endometrial cancer, adrenocortical cancer, uterine cancer, cervical cancer, esophageal cancer, breast cancer, kidney cancer, and ovarian cancer.

[0039] Terminology Definition

[0040] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0041] In the compounds of this invention, "hydrogen," "carbon," and "oxygen" include all their isotopes. Isotopes should be understood to include those atoms having the same number of atoms but different mass numbers. For example, isotopes of hydrogen include protium, tritium, and deuterium, and isotopes of carbon include... 12 C 13 C and 14 C, oxygen isotopes include 16 O and 18 O etc.

[0042] In this invention, "isomers" refers to molecules with the same atomic composition and bonding but different three-dimensional spatial arrangements, including but not limited to diastereomers, enantiomers, cis-trans isomers, and mixtures thereof, such as racemic mixtures. Many organic compounds exist in optically active forms, meaning they are capable of rotating the plane of polarized light. In describing optically active compounds, the prefixes D, L, or R, S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes D, L, or (+), (-) are used to name the symbols for the plane polarization rotation of the compound; (-) or L indicates that the compound is levorotatory, and the prefix (+) or D indicates that the compound is dextrorotatory. These stereoisomers have the same chemical structure but different stereostructures. Specific stereoisomers can be enantiomers, and mixtures of isomers are usually called enantiomer mixtures. A 50:50 enantiomer mixture is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereodirection during chemical reactions. The terms “racemic mixture” and “racemate” refer to a mixture of two equimolar enantiomers that lack optical activity.

[0043] Depending on the choice of starting materials and methods, the compounds of this invention can exist as one or a mixture of possible isomers, such as racemic mixtures and mixtures of non-corresponding isomers (depending on the number of asymmetric carbon atoms). Optically active (R)- or (S)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

[0044] Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0045] In this invention, "halogen" refers to fluorine, chlorine, bromine, or iodine. "Halogenated" in this invention means substituted with fluorine, chlorine, bromine, or iodine.

[0046] In this invention, "alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group, preferably a straight-chain or branched group containing 1 to 6 carbon atoms, and more preferably a straight-chain or branched group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc. The alkyl group can be substituted or unsubstituted, and when substituted, the substituent can be at any usable connection point.

[0047] In this invention, "carbonyl" and "acyl" both refer to -C(O)-.

[0048] In this invention, "sulfonyl" refers to -S(O)2-.

[0049] In this invention, "sulfonamide group" refers to -S(O)2NH-.

[0050] In this invention, "halogenated alkyl" refers to an alkyl group that is substituted with at least one halogen.

[0051] In this invention, "hydroxyalkyl" refers to an alkyl group that is substituted with at least one hydroxyl group.

[0052] In this invention, "alkoxy" refers to -O-alkyl. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy, etc. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any usable connection point.

[0053] In this invention, "cycloalkyl" refers to a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups can be substituted or unsubstituted monocyclic, bicyclic, or tricyclic saturated hydrocarbon groups having 3-12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0054] The term "heterocyclic group" in this invention refers to a group having a 3- to 15-membered non-aromatic ring system ("3- to 15-membered heterocyclic group") having 1 to 4 ring heteroatoms (each heteroatom independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon). In a heterocyclic group containing one or more nitrogen atoms, the connecting point can be a carbon or nitrogen atom, provided the valence permits. The heterocyclic group can be monocyclic ("monocyclic heterocyclic group") or a fused, bridged, or spirocyclic ring system (e.g., a bicyclic system (also called "bicyclic heterocyclic group")) and can be saturated or partially unsaturated, wherein the bicyclic heterocyclic group includes, but is not limited to, benzo[a]azine heterocyclic group, benzo[ox]oxine heterocyclic group, benzo[thio]sulfine heterocyclic group, benzo[diazepine]azine heterocyclic group, benzo[dioxane]oxine heterocyclic group, benzo[disulfine]sulfine heterocyclic group, benzo[oxane]azine heterocyclic group, and benzo[thio]sulfine heterocyclic group. Suitable heterocyclic groups include, but are not limited to, piperidinyl, aziridine, aziridine propane, tetrahydropyrrole, piperazine, dihydroquinazolinyl, oxetanepropyl, oxetanebutyl, tetrahydrofuranyl, tetrahydropyranyl, dihydrobenzoxycyclohexenyl, dihydroquinolinyl, tetrahydroquinolinyl, dihydroisoquinolinyl, and tetrahydroisoquinolinyl. Each example of a heterocyclic group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any usable connection point.

[0055] In this invention, "aryl" refers to an aromatic system that may comprise a monocyclic or fused polycyclic ring, preferably a monocyclic or fused bicyclic aromatic system, containing 6 to 12 carbon atoms, more preferably about 6 to about 10 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthraceneyl, fluorenyl, and indanyl. The aryl group may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any usable connection point.

[0056] In this invention, "heteroaryl" refers to an aryl group in which at least one carbon atom is replaced by a heteroatom, preferably consisting of 5-12 atoms (5-12-membered heteroaryl), more preferably consisting of 5-10 atoms (5-10-membered heteroaryl), wherein the heteroatom is O, S, or N. The heteroaryl groups include, but are not limited to, imidazolyl, pyrrolyl, furanyl, thiophene, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, indolyl, pyridyl, pyrimidinyl, pyrazinyl, triazinyl, isoindolyl, benzopyrazolyl, benzoimidazolyl, benzofuranyl, benzopyranyl, benzothiophene, benzooxazolyl, benzothiazolyl, benzothiazolyl, benzoisooxazolyl, benzoisothiazolyl, quinolinyl, isoquinolinyl, quinazolinyl, cinolinyl, quinoxazinyl, benzothiazolyl, imidazopyridyl, pyrimidinylpyrazolyl, pyrimidinylimidazolyl, pyrazinylpyrroleyl, pyrazinylpyrroleyl, pyridinylpyrroleyl, pyridinylpyrazolyl, etc. The heteroaryl group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any usable connection point.

[0057] In this invention, "cycloalkylalkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group has been replaced by a cycloalkyl group as defined herein.

[0058] In this invention, "heterocyclic alkoxy" refers to an alkoxy group in which at least one hydrogen atom of the alkoxy group has been replaced by a heterocyclic group as defined herein.

[0059] The term "pharmaceutically acceptable salt" in this invention refers to salts of the compounds of this invention that are safe and effective when used in mammals and possess the intended biological activity.

[0060] In the conventional sense, the term "solvent" in this invention refers to a complex formed by a combination of a solute (such as an active compound or a salt of an active compound) and a solvent (such as water). The solvent refers to a solvent known or readily identifiable to those skilled in the art. If water is present, the solvate is typically referred to as a hydrate, such as a hemihydrate, monohydrate, dihydrate, trihydrate, or a substitute thereof.

[0061] The in vivo effects of compounds having chemical formula (I) can be partially exerted by one or more metabolites formed in the human or animal body after administration of the compound having chemical formula (I). As described above, the in vivo effects of compounds having chemical formula (I) can also be exerted via the metabolism of a prodrug ("prodrug"). The "prodrug" of the present invention refers to a compound that, under physiological conditions in an organism, is converted into the compound of the present invention through reaction with enzymes, gastric acid, etc., i.e., a compound converted into the compound of the present invention through enzymatic oxidation, reduction, hydrolysis, etc., and / or through hydrolytic reactions such as gastric acid, etc.

[0062] The “crystallization” of this invention refers to a solid whose internal structure is formed by the regular repetition of atoms (or groups thereof) in three dimensions, which is different from amorphous solids that do not have such a regular internal structure.

[0063] The term "pharmaceutical composition" as used in this invention refers to a mixture comprising any of the compounds described herein, including corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts or their chemically protected forms, and one or more pharmaceutically acceptable carriers and / or mixtures of other one or more drugs. The purpose of a pharmaceutical composition is to facilitate the administration of the compound to a living organism. Such compositions are typically used in the preparation of medicaments for the treatment and / or prevention of diseases mediated by one or more kinases.

[0064] The "pharmaceutical-grade carrier" of this invention refers to a carrier that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the administered compound. This includes all solvents, diluents or other excipients, dispersants, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc., unless any conventional carrier medium is incompatible with the compounds of this invention. Some examples of pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, as well as cellulose and cellulose acetate; malt, gelatin, etc.

[0065] In this invention, "excipient" refers to an inert substance added to a pharmaceutical composition to further promote the delivery of the compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. Detailed Implementation

[0066] The present invention will be further described in detail below with reference to the embodiments, but the present invention is not limited to these embodiments. Unless otherwise specified, all materials used in the following embodiments are commercially available.

[0067] Intermediate 1: (S,E)-1-cyclopropyl-3-(methanesulfonyl)propyl-2-en-1-amine p-toluenesulfonate

[0068]

[0069] Step 1: Preparation of (S)-(1-cyclopropyl-2-hydroxyethyl)carbamate tert-butyl ester

[0070]

[0071] (S)-2-((tert-Butoxycarbonyl)amino)-2-cyclopropylacetic acid (2.25 g, 10 mmol) was dissolved in tetrahydrofuran (40 mL), followed by the addition of triethylamine (1.38 mL, 10 mmol). The mixture was stirred in an ice-water bath for 15 min, and then 5 mL of a tetrahydrofuran solution of isobutyl chloroformate (1.36 mL, 10 mmol) was added dropwise. The mixture was stirred in an ice-water bath for 1 h, followed by filtration. A 20% tetrahydrofuran aqueous solution of sodium borohydride was added to the filtrate in an ice-water bath, and the mixture was stirred for another 2 h. The reaction was quenched by adjusting the pH to 4-5 with dilute hydrochloric acid, and then the pH was adjusted to 7-8 with a saturated sodium bicarbonate aqueous solution. The mixture was extracted with ethyl acetate, and the organic phases were combined, dried, and concentrated to give 1.91 g of a transparent oil. ESI-MS m / z: 146.1 [M+H-56] + .

[0072] Step 2: Preparation of (S)-(1-cyclopropyl-2-oxoethyl)carbamate tert-butyl ester

[0073]

[0074] Dissolve Dys-Martin oxidant (8.15 g, 19.0 mmol) in dichloromethane (40 mL). Stir for 15 min in an ice-water bath, then add a dichloromethane solution of (S)-(1-cyclopropyl-2-hydroxyethyl)carbamate tert-butyl ester (1.91 g, 9.5 mmol). After the addition is complete, transfer the mixture to room temperature and react for 3 h 10 min later. Filter the reaction mixture, wash the filtrate with saturated sodium chloride solution, and extract with ethyl acetate. Combine the organic phases, dry them, and purify by column chromatography to obtain 1.03 g of a clear oil. ESI-MS m / z: 144.1 [M+H-56] + .

[0075] Step 3: Preparation of (S,E)-(1-cyclopropyl-3-(methylsulfonyl)allyl)carbamate tert-butyl ester

[0076]

[0077] Diethyl ((methanesulfonyl)methyl)phosphonate (1.18 g, 5.12 mmol) was dissolved in tetrahydrofuran (20 mL), and stirred at -30 °C for 15 min. Sodium hydride (400 mg, 10 mmol) was added, and the mixture was stirred for 1 h. Then, a tetrahydrofuran solution of (S)-(1-cyclopropyl-2-oxoethyl)carbamate tert-butyl (1.03 g, 5.12 mmol) was added dropwise, followed by stirring at -30 °C for 2–3 h. The reaction was quenched with water at low temperature, and the mixture was extracted with ethyl acetate. The organic phases were combined, dried, and purified by column chromatography to give 600 mg of an oily substance. ESI-MS m / z: 220.0 [M+H-56] + .

[0078] Step 4: Preparation of (S,E)-1-cyclopropyl-3-(methanesulfonyl)propyl-2-en-1-amine p-toluenesulfonate

[0079]

[0080] (S,E)-(1-cyclopropyl-3-(methanesulfonyl)allyl)carbamate tert-butyl ester (600 mg, 2.18 mmol) was dissolved in acetonitrile (15 mL), and p-toluenesulfonic acid (450.5 mg, 2.62 mmol) was added. The mixture was transferred to 50 °C and reacted overnight. LC-MS was used to confirm the completeness of the reaction, and the reaction mixture was concentrated for later use. ESI-MS m / z: 176.0 [M+H] +.

[0081] Example 1: N-((1S,E)-1-cyclopropyl-3-(S-methylsulfonylimino)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0082]

[0083] Step 1: Preparation of methyl 4-bromo-5-fluoro-2-phenoxybenzoate

[0084]

[0085] Methyl 4-bromo-2,5-difluorobenzoate (5 g, 18.7 mmol) and phenol (2.1 g, 22.4 mmol) were added to a solution of N,N-dimethylformamide (10 mL), followed by the addition of N,N-diisopropylethylamine (4.8 g, 37.4 mmol), and then... o The mixture was stirred at C for 5 hours. It was then extracted with water and ethyl acetate, and the organic phase was passed through a silica gel column to obtain the product. ESI-MS m / z: 325.0 [M+H] + .

[0086] Step 2: Preparation of methyl 4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzoate

[0087]

[0088] Methyl 4-bromo-5-fluoro-2-phenoxybenzoate (20 g, 61 mmol), sodium cyclopropyl sulfinate (12 g, 91.5 mmol), cuprous iodide (2.3 g, 12.2 mmol), and L-proline (1.3 g, 12.2 mmol) were dissolved in dimethyl sulfoxide. The mixture was stirred at 100 °C for 6 hours under argon protection. After the reaction was complete, water was added to quench the reaction mixture, followed by extraction with ethyl acetate, backwashing with water, washing with saturated sodium chloride, concentration under reduced pressure, and chromatography on silica gel to obtain the product. ESI-MS m / z: 350.9 [M+H] + .

[0089] Step 3: Preparation of 4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzoic acid

[0090]

[0091] Methyl 4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzoate (2 g, 5.7 mmol) was dissolved in tetrahydrofuran and methanol, and then an aqueous solution of 6N sodium hydroxide was added. After stirring for 5 hours, the mixture was evaporated to dryness to obtain the product. ESI-MS m / z: 337.0 [M+H]+ .

[0092] Step 4: Preparation of (S)-N-(1-cyclopropyl-2-hydroxyethyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0093]

[0094] 4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzoic acid (1.9 mg, 5.7 mmol), (S)-2-amino-2-cyclopropylethane-1-ol (575 mg, 5.7 mmol), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (2.2 g, 5.7 mmol), and N,N-diisopropylethylamine (200 mg) were dissolved in N,N-dimethylformamide. The mixture was stirred at room temperature for 5 hours, extracted with water and ethyl acetate, washed with saturated sodium chloride, concentrated under reduced pressure, and column chromatography to obtain the product. ESI-MS m / z: 419.9 [M+H] + .

[0095] Step 5: Preparation of (S)-N-(1-cyclopropyl-2-oxoethyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0096]

[0097] (S)-N-(1-cyclopropyl-2-hydroxyethyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide (2.0 g, 5.0 mmol) was dissolved in a 1:1 mixture of acetonitrile and dichloromethane. Dys-Martin reagent (4.24 g, 10.0 mmol) was added in an ice bath, and the mixture was stirred at room temperature for 2 hours. The solution was then concentrated by filtration and column chromatography to obtain the product. ESI-MS m / z: 417.9 [M+H] + .

[0098] Step 6: Preparation of (S,E)-N-(1-cyclopropyl-3-(methylthio)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0099]

[0100] 40 mL of tetrahydrofuran was added to (methylthio)methyl)triphenylphosphine hydrochloride (1.5 g, 4.1 mmol) under argon protection. 2 mL of n-butyllithium (2.5 mol / L in n-hexane) was added at -20 °C. After stirring for half an hour, (S)-N-(1-cyclopropyl-2-oxoethyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide (860 mg, 2.1 mmol) was added. The mixture was stirred at -20 °C for 2 hours, quenched with water, extracted with ethyl acetate, backwashed with water, washed with saturated sodium chloride, concentrated under reduced pressure, and column chromatography to obtain the product. ESI-MS m / z: 461.9 [M+H] + .

[0101] Step 7: Preparation of N-((1S,E)-1-cyclopropyl-3-(S-methylsulfonylimino)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0102]

[0103] (S,E)-N-(1-cyclopropyl-3-(methylthio)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide (650 mg, 1.4 mmol) was dissolved in methanol, and ammonium acetate (323 mg, 4.2 mmol) and iodophenylacetic acid (681 mg, 2.1 mmol) were added. After stirring for 5 hours, the mixture was evaporated to dryness to obtain the product. ESI-MS m / z: 492.9 [M+H] + .

[0104] 1 H NMR (400 MHz, DMSO-d6) δ 8.93 (t, J = 8.0 Hz, 1H), 7.83 (d, J =9.4 Hz, 1H), 7.45 (t, J = 7.9 Hz, 2H), 7.26 – 7.20 (m, 2H), 7.14 (d, J = 8.0Hz, 2H), 6.79 - 6.67 (m, 2H), 4.20 - 4.12 (m, 1H), 3.06-2.99 (m, 1H), 2.82(d, J = 2.8 Hz, 3H), 1.13 (d, J = 6.3 Hz, 4H), 1.04 - 0.94 (m, 1H), 0.53-0.48(m, 1H), 0.44 – 0.33 (m, 2H), 0.32 – 0.22 (m, 1H).

[0105] Example 2: N-((1S,E)-1-cyclopropyl-3-(S-methylsulfonylimino)allyl)-4-(trifluoromethylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0106]

[0107] Step 1: Preparation of methyl 5-fluoro-2-phenoxy-4-((trifluoromethyl)thio)benzoate

[0108]

[0109] Methyl 4-bromo-5-fluoro-2-phenoxybenzoate (1.5 g, 4.6 mmol) was dissolved in 10 mL of N-methylpyrrolidone, followed by the addition of copper trifluoromethylthiocyanate (2.3 g, 13.9 mmol). The mixture was stirred in a microwave oven at 150 °C for 9 hours, and the reaction was monitored by LC-MS until completion. The product was obtained by column chromatography. ESI-MS m / z: 346.8 [M+H] + .

[0110] Step 2: Preparation of methyl 5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzoate

[0111]

[0112] Methyl 5-fluoro-2-phenoxy-4-((trifluoromethyl)thio)benzoate (300 mg, 0.867 mmol) was dissolved in 10 mL dichloromethane / 10 mL acetonitrile / 20 mL water, followed by the addition of sodium periodate (556 mg, 2.6 mmol). The mixture was then cooled to 0 °C, and ruthenium trichloride monohydrate (4 mg, 0.017 mmol) was added. The mixture was stirred at room temperature for 2 hours, and the reaction was monitored by LC-MS until completion. The product was obtained by column chromatography. ESI-MS m / z: 378.9 [M+H] + .

[0113] Step 3: Preparation of 5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzoic acid

[0114]

[0115] Methyl 5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzoate (380 mg, 1.0 mmol) was dissolved in 20 mL of tetrahydrofuran and 20 mL of water, followed by the addition of lithium hydroxide (72 mg, 3.0 mmol). The mixture was stirred at room temperature for 2 hours, and the reaction was monitored by LC-MS until completion. The pH of the aqueous phase was adjusted to approximately 3.0, and the product was extracted with ethyl acetate and concentrated to obtain the final product. ESI-MS m / z: 364.8 [M+H] + .

[0116] Step 4: Preparation of (S)-N-(1-cyclopropyl-2-hydroxyethyl)-5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzamide

[0117]

[0118] The preparation method is the same as that in Example 1, except that the starting material 5-fluoro-2-phenoxy-4-(cyclopropylsulfonyl)benzoic acid is replaced with 5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzoic acid, thus obtaining the title compound. ESI-MS m / z: 448.1 [M+H] + .

[0119] Step 5: Preparation of (S)-N-(1-cyclopropyl-2-oxoethyl)-5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzamide

[0120]

[0121] The preparation method was the same as in Example 1, except that the starting material (S)-N-(1-cyclopropyl-2-hydroxyethyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide was replaced with (S)-N-(1-cyclopropyl-2-hydroxyethyl)-5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzamide to obtain the title compound. ESI-MS m / z: 446.1 [M+H] + .

[0122] Step 6: Preparation of (S,E)-N-(1-cyclopropyl-3-(methylthio)allyl)-4-((trifluoromethyl)sulfonyl)-5-fluoro-2-phenoxybenzamide

[0123]

[0124] The preparation method was the same as in Example 1, except that the starting material (S)-N-(1-cyclopropyl-2-oxoethyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide was replaced with (S)-N-(1-cyclopropyl-2-oxoethyl)-5-fluoro-2-phenoxy-4-((trifluoromethyl)sulfonyl)benzamide to obtain the title compound. ESI-MS m / z: 490.1 [M+H] + .

[0125] Step 7: Preparation of N-((1S,E)-1-cyclopropyl-3-(S-methylsulfonylimino)allyl)-4-(trifluoromethylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0126]

[0127] The preparation method was the same as in Example 1, except that the starting material (S,E)-N-(1-cyclopropyl-3-(methylthio)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide was replaced with (S,E)-N-(1-cyclopropyl-3-(methylthio)allyl)-4-((trifluoromethyl)sulfonyl)-5-fluoro-2-phenoxybenzamide to obtain the title compound. ESI-MS m / z: 521.1 [M+H] + .

[0128] 1 H NMR (400 MHz, DMSO-d6) δ 9.03 (t, J = 7.9 Hz, 1H), 8.07 (d, J =9.7 Hz, 1H), 7.54 – 7.35 (m, 3H), 7.25 (t, J = 7.3 Hz, 1H), 7.16 (d, J = 7.8Hz, 2H), 6.81 – 6.64 (m, 2H), 4.20 – 4.11 (m, 1H), 3.86 (d, J = 6.3 Hz, 1H), 2.82 (d, J = 2.4 Hz, 3H), 1.04 – 0.93 (m, 1H), 0.55-0.48 (m, 1H), 0.45 – 0.33(m, 2H), 0.31 – 0.22 (m, 1H).

[0129] Example 3: N-((1S,E)-1-cyclopropyl-3-(N,S-dimethylsulfonylimino)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0130]

[0131] N-((1S,E)-1-cyclopropyl-3-(S-methylsulfonylimino)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide (200 mg, 0.4 mmol) was dissolved in 10 mL of 1,4-dioxane, followed by the addition of trimethylcycloboroxane (0.06 mL, 0.5 mmol), copper acetate (0.09 g, 0.5 mmol), and pyridine (0.1 mL, 1.2 mmol). The mixture was purged with argon three times, and the reaction was stirred at 100 °C. The reaction was monitored by LC-MS until completion. The product was obtained by extraction with ethyl acetate, backwashing with water, drying on anhydrous sodium sulfate, silica gel column chromatography, and HPLC. ESI-MS m / z: 507.1 [M+H] + .

[0132] 1 H NMR (400 MHz, CDCl3) δ 8.09 (d, J = 10.0 Hz, 1H), 7.92 (d, J = 7.0Hz, 1H), 7.47 – 7.43 (m, 2H), 7.26 (s, 3H), 7.29 – 7.26 (m, 1H), 6.88 – 6.80(m, 1H), 6.25 (d, J = 14.9 Hz, 1H), 4.24 – 4.22 (m, 1H), 2.95 – 2.94 (m, 2H), 2.75 – 2.68 (m, 3H), 1.67 – 1.56 (m, 3H), 1.39 – 1.21 (m, 3H), 1.12 – 0.09(m, 1H), 0.98 (s, 1H), 0.66 – 0.64 (m, 1H), 0.58 – 0.48 (m, 1H), 0.40 – 0.39(s, 1H).

[0133] Example 4: N-((1S,E)-1-cyclopropyl-3-(N,S-dimethylsulfonylimino)allyl)-4-(trifluoromethylsulfonyl)-5-fluoro-2-phenoxybenzamide

[0134]

[0135] The preparation method was the same as in Example 3, except that the starting material N-((1S,E)-1-cyclopropyl-3-(S-methylsulfonylimino)allyl)-4-(cyclopropylsulfonyl)-5-fluoro-2-phenoxybenzamide was replaced with N-((1S,E)-1-cyclopropyl-3-(S-methylsulfonylimino)allyl)-4-(trifluoromethylsulfonyl)-5-fluoro-2-phenoxybenzamide to obtain the title compound. ESI-MS m / z: 535.1 [M+H] + .

[0136] 1 H NMR (400 MHz, DMSO-d6) δ 9.05 (d, J = 8.2 Hz, 1H), 8.06 (d, J = 9.6Hz, 1H), 7.51 – 7.37 (m, 3H), 7.29 – 7.21 (m, 1H), 7.15 (d, J = 8.0 Hz, 2H),6.69 – 6.61 (m, 1H), 6.52 – 6.43 (m, 1H), 4.24 – 4.12 (m, 1H), 2.89 – 2.83(m, 3H), 1.23 (s, 3H), 1.08 – 0.94 (m, 1H), 0.56 – 0.46 (m, 1H), 0.45 – 0.34(m, 2H), 0.33 – 0.24(m, 1H).

[0137] Example 5: (S,E)-N-(1-cyclopropyl-3-(methylsulfonyl)allyl)-6-methyl-5-((trifluoromethyl)sulfonyl)-3-phenoxypyridinecarboxamide

[0138]

[0139] The preparation method was the same as in Example 2, except that methyl 4-bromo-5-fluoro-2-phenoxybenzoate was replaced with methyl 5-bromo-3-phenoxy-6-methylpyridinecarboxylate, and (S)-2-amino-2-cyclopropylethane-1-ol was replaced with (S,E)-1-cyclopropyl-3-(methanesulfonyl)propyl-2-en-1-amine p-toluenesulfonate, to obtain the title compound. ESI-MS m / z: 519.1 [M+H] + .

[0140] 1H NMR (400 MHz, DMSO-d6) δ 9.14 (s, 1H), 9.12 (d, J = 8.5 Hz, 1H), 7.33 (t, J = 8.0 Hz, 2H), 7.09 (t, J = 7.4 Hz, 1H), 6.89 (d, J = 8.3 Hz, 2H),6.69 (s, 1H), 6.63 (d, J = 15.4 Hz, 1H), 4.01-3.79 (m, 1H), 2.96 (s, 3H),2.47 (s, 3H), 1.01 - 0.94 (m, 1H), 0.52- 0.46 (m, 1H), 0.39 - 0.31 (m, 2H),0.18 - 0.13 (m, 1H).

[0141] Following the synthesis methods described in the above embodiments, compounds from Examples 6-14 in Table 1 were prepared using different raw materials.

[0142] Table 1

[0143]

[0144] Experimental Example 1: Evaluation of the in vitro cell activity of compounds

[0145] 1. Experimental Materials

[0146] Experimental cells: Human colorectal cancer cells HCT116 were purchased from Wuhan Pronosei Life Science Technology Co., Ltd.

[0147] Test compounds: The compounds of the present invention prepared in the above examples were each prepared into a 20 mM stock solution using DMSO. The drugs were diluted with complete culture medium, and the final drug concentration was started at 5 μM as the highest concentration. The drugs were then diluted sequentially in a 1:3 ratio to nine concentration gradients: 5 μM, 1.25 μM, 1.11 μM, 0.313 μM, 0.0195 μM, 0.00488 μM, 0.00122 μM, 0.000305 μM, and 0.000076 μM.

[0148] Reagents: McCoy's 5A (Gibco, 16600-082); Fetal Bovine Serum (Gibco, 10099-141); Penicillin and Streptomycin (Gibco, 15140-122); CellTiter-Glo Luminescent Cell ViabilityAssay (Promega, G7571).

[0149] 2. Experimental Methods

[0150] 2.1 Cell resuscitation:

[0151] Remove the HCT116 cell cryovials from the liquid nitrogen tank and place them at 37°C. o In a C2 water bath, gently agitate to thaw quickly. After thawing, remove the cryovials, sterilize with an alcohol swab, unscrew the cap, aspirate the cell suspension into a centrifuge tube, add 1 mL of McCoy's 5A medium containing 10% FBS, mix well, and centrifuge at 1000 rpm for 5 min. Discard the supernatant and resuspend the cells in complete culture medium. Seed the cells at an appropriate concentration in culture dishes. Incubate at 37°C. o Incubate in a 5% CO2 incubator (C).

[0152] 2.2 Cell Culture and Passaging:

[0153] When cells reach approximately 80-90% confluence, discard the original culture medium (McCoy's 5A medium + 10% FBS + 1% penicillin and streptomycin). Wash away any remaining medium with 1 mL of PBS and discard the PBS. Add 1 mL of trypsin digest and digest for 2-3 minutes. Observe under a microscope that the pseudopodia of the cells have retracted and become rounded, but the cells have not yet detached in sheets. At this point, discard the trypsin and terminate digestion with 1-2 mL of complete culture medium. Gently pipette and collect the cell suspension. Centrifuge at 1000 rpm for 5 minutes. Remove the supernatant, resuspend the cells in complete culture medium, and seed them in culture dishes at the desired density. Incubate at 37°C. o Cultured in a 5% CO2 incubator, the culture medium was changed or passaged every 2-3 days depending on cell growth.

[0154] 2.3 Experimental Procedure:

[0155] Day 1 of the experiment:

[0156] After passage, HCT116 cells were resuspended in complete culture medium at the appropriate density and seeded into 96-well plates: the outermost 36 wells were filled with 200 μL of PBS to prevent excessive differences in culture conditions between the inner wells due to rapid evaporation of the medium at the edge; the remaining 60 wells were seeded; each well contained 100 μL of complete culture medium with 750 cells, with two replicates for each concentration. The plates were then placed in a 5% CO2 incubator at 37°C. o Cultured for 24 hours.

[0157] Day 2 of the experiment:

[0158] HCT116 cells were cultured in a 5% CO2 incubator for 4 days after adding 100 μL of (2x) drug to the original culture medium (100 μL). Two replicates were set for each concentration group.

[0159] Day 5 of the experiment:

[0160] Four days after HCT116 cell treatment, the CellTiter-Glo Luminescent CellViabillity Assay was removed 30 min in advance and allowed to equilibrate to room temperature. The culture medium in the wells was discarded, and Celltiter-Glo reagent was diluted 1:1 (equal volume) with complete culture medium. 150 μL of the diluted Celltiter-Glo reagent was added to each well, and the cells were shaken at room temperature for 2 min. After incubation at room temperature for another 10 min, 100 μL of the chemiluminescence signal was taken from each well for detection, followed by shaking. The read detection condition was 500 ms. Based on the AU value exported from the microplate reader, the inhibition rate of each well relative to the solvent control well was calculated: Inhibition (%) = 100 - (AU experimental wells - AU blank wells) / (AU solvent control wells - AU blank wells) * 100. Based on different drug concentrations and their corresponding inhibition rates, a four-parameter logistic sigmoid curve was fitted using GraghPad 6.0 software, and the data were analyzed to derive the IC50. 50 Values. The experimental results are shown in Table 2.

[0161] Table 2

[0162]

[0163] Experimental results show that the compound of the present invention has good inhibitory activity against the proliferation of human colorectal cancer cells HCT116.

[0164] Although the present invention has been described in detail above, those skilled in the art will understand that various modifications and changes can be made to the invention without departing from its spirit and scope. The scope of the invention is not limited to the detailed description above, but should be attributed to the claims.

Claims

1. A compound of general formula (I) or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, in, X is selected from CH and N; Q is selected from =O, =NH, and =NCH3; W is selected from halogen, cyano, amino, alkyl, cycloalkyl, heterocyclic, haloalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, alkylamino, aryl, heteroaryl, arylalkyl, heteroarylalkyl, cycloalkylalkyl, and heterocyclicalkyl, optionally substituted by one or more groups selected from halogen, hydroxy, amino, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, aryl, heteroaryl, cycloalkyl, and heterocyclicalkyl; R 1 Selected from hydrogen, halogen, cyano, amino, alkyl, oxo group, cycloalkyl, heterocyclic, heterocyclic alkenyl, haloalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, monoalkylamino, dialkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, aryl, and heteroaryl; and R 2 It is selected from alkyl, haloalkyl, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclic, aryl and heteroaryl groups, and is optionally substituted by one or more groups selected from halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, amino, monoalkylamino, alkylacylamino, alkylacyl, alkylsulfonyl, aminoacyl, alkylaminoacyl, dialkylamino, haloalkylacyl, hydroxyalkylacyl, cycloalkyl, heterocyclic, aryl and heteroaryl groups.

2. The compound according to claim 1 or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, wherein W is selected from halogen, cyano, amino, C 1-6 Alkyl, C 3-10 cycloalkyl, C 3-10 Heterocyclic groups, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, C 1-6 Alkylamino, C 6-10 Aryl, 5-10 heteroaryl, C 6-10 Aryl C 1-6 Alkyl, 5-10-membered heteroaryl C 1-6 Alkyl, C 3-10 cycloalkyl C 1-6 Alkyl and C 3-10 Heterocyclic C 1-6 Alkyl groups, optionally composed of one or more elements selected from halogens, hydroxyl groups, amino groups, C4 groups, and hydroxyl groups. 1-6 Alkyl, Halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, C 6-10 Aryl, 5-10 heteroaryl, C 3-10 cycloalkyl and C 3-10 The heterocyclic alkyl group is substituted, wherein the heteroatom is selected from N, O, S and P.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, wherein R 1 Selected from hydrogen, halogen, cyano, amino, C 1-6 Alkyl, oxo group, C 3-10 Cycloalkyl, 3-10 heterocyclic, 3-10 heterocyclic alkenyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, single C 1-6 Alkylamino, bis(C) 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, bis(C) 1-6 Alkylamino, C 6-10 Aryl and 5-10 heteroaryl groups, wherein the heteroatom is selected from N, O, S and P.

4. The compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, wherein R 2 Selected from C 1-6 Alkyl, Halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, C 3-10 Cycloalkyl, 3-10 membered heterocyclic groups, C 6-10 aryl and 5-10 heteroaryl groups, optionally separated by one or more elements selected from halogen, hydroxyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, amino, C 1-6 Monoalkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, bis(C) 1-6 Alkylamino, halogenated C 1-6 Alkyl acyl, hydroxy C 1-6 Alkyl acyl, C 3-8 Cycloalkyl, 3-8 membered heterocyclic groups, C 6-12 The aryl group is substituted with a 5-12-membered heteroaryl group, wherein the heteroatom is selected from N, O, S and P.

5. A compound of general formula (II) or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, Where Y is selected from O and NH, and n is selected from 0, 1, 2, 3 and 4.

6. The compound according to claim 1 or 5, or a pharmaceutically acceptable salt, isomer, solvate, crystal, or prodrug thereof, wherein the compound is selected from the group consisting of: ， ， ， ， ， ， ， ， ， ， ， ， ,and .

7. A pharmaceutical composition comprising the compound of any one of claims 1-6 or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug and a pharmaceutically acceptable carrier.

8. The use of the compound of any one of claims 1-6 or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug or the pharmaceutical composition of claim 7 in the preparation of a medicament for treating diseases associated with WRN.

9. The application according to claim 8, wherein the disease is cancer.

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