Short peptide compounds containing beta-amino ketones and uses thereof

Abstract

A kind of short peptide compound containing beta-amino ketone and its application, specifically disclose the compound shown in formula (II) or its pharmaceutically acceptable salt.

Description

[0001] This application claims the following priority:

[0002] Application number: CN2021115809216, application date: December 22, 2021;

[0003] Application number: CN2022100451335, application date: January 14, 2022;

[0004] Application number: CN2022102216236, application date: March 8, 2022;

[0005] Application number: CN2022110234109, application date: August 24, 2022. Technical Field

[0006] This invention relates to the field of pharmaceutical chemistry, and in particular to a short peptide compound containing a β-amino ketone and its applications. Background Technology

[0007] (3CL pro 3CL proteases belong to the cysteine ​​protease family. Their substrate-binding sites are highly conserved and they possess similar catalytic mechanisms, making them key proteases for catalyzing the cleavage of RNA virus precursor proteins and playing a crucial role in viral replication. As a key enzyme in coronavirus replication, 3CL protease (3CL...) pro 3CL is an important target for treating diseases caused by various coronaviruses, including COVID-19. Utilizing the structural characteristics of the 3CL protease to develop drugs for treating COVID-19 infection has significant clinical value. Summary of the Invention

[0008] This invention provides compounds of formula (II) or pharmaceutically acceptable salts thereof.

[0009]

[0010] in,

[0011] R1 is independently selected from halogens, CN, OH, NH2, and C. 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl or C 1-3 Each alkoxy group can be independently substituted by one, two, or three halogens;

[0012] or,

[0013] Two R1 atoms and the atoms bonded to them form C 3-6 cycloalkyl, the C 3-6 The cycloalkyl group is optionally surrounded by 1, 2 or 3 Rs. a replace;

[0014] R a Each is independently selected from halogens and C. 1-3 alkyl;

[0015] n is selected from 0, 1, 2, or 3;

[0016] R2 is selected from tert-butyl, C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl and phenyl, wherein C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl and phenyl groups are each independently optionally bounded by 1, 2 or 3 R groups. b replace;

[0017] R b Each is independently selected from halogens and C. 1-3 alkyl;

[0018] R3 is selected from C 1-3 Alkyl, C 1-3 Alkoxy, -CH2R4 and -CH2OR4, the C 1-3 Alkyl and C 1-3 Each alkoxy group can be independently substituted by one, two, or three halogens;

[0019] R4 is selected from phenyl and 5-6 heteroaryl groups, each of which is independently optionally substituted by 1, 2 or 3 R groups;

[0020] R is selected from halogen and C. 1-3 alkyl;

[0021] T1 is selected from O and S;

[0022] Ring A is selected from

[0023] A1, A2, A3, and A4 are each independently selected from CH and N.

[0024] In some embodiments of the present invention, R1 is selected from F and methyl, and other variables are as defined in the present invention.

[0025] In some embodiments of the present invention, the two R1s and the atoms connected to them form C. 3-6 Cycloalkyl groups, and other variables as defined in this invention.

[0026] In some embodiments of the present invention, the two R1s and the atoms connected to them form a cyclopropyl group, and other variables are as defined in the present invention.

[0027] In some embodiments of the present invention, the above-mentioned structural unit Selected from Other variables are as defined in this invention.

[0028] In some embodiments of the present invention, R2 is selected from tert-butyl and adamantyl, wherein the adamantyl group is optionally surrounded by 1, 2, or 3 R's. b Replacement, other variables as defined in this invention.

[0029] In some embodiments of the present invention, R2 is selected from tert-butyl and adamantyl, and other variables are as defined in the present invention.

[0030] In some embodiments of the present invention, R2 is selected from tert-butyl, Other variables are as defined in this invention.

[0031] In some embodiments of the present invention, the above-mentioned structural unit Selected from Other variables are as defined in this invention.

[0032] In some embodiments of the present invention, R3 is selected from -CF3, and other variables are as defined in the present invention.

[0033] The present invention also provides compounds shown in (II-1), (II-2) and (II-3) or pharmaceutically acceptable salts thereof.

[0034]

[0035] Where t is selected from 0, 1, and 2;

[0036] R1, R b A1, A2, A3, A4, T1, and n are as defined in this invention.

[0037] The present invention also provides compounds of formula (I) or pharmaceutically acceptable salts thereof.

[0038]

[0039] in,

[0040] R1 is independently selected from halogens, CN, OH, NH2, and C. 1-3 Alkyl or C 1-3 Alkoxy, the C 1-3 Alkyl or C 1-3 The alkoxy groups can be independently replaced by one, two, or three halogens;

[0041] or,

[0042] Two R1 atoms and the atoms bonded to them form C 3-6 cycloalkyl, the C 3-6 The cycloalkyl group is optionally surrounded by 1, 2 or 3 Rs. a replace;

[0043] n is selected from 0, 1, 2, or 3;

[0044] R a Each is independently selected from halogen or C 1-3 alkyl;

[0045] R2 is selected from tert-butyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl or phenyl, wherein C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl and phenyl groups are optionally surrounded by 1, 2 or 3 R groups. b replace;

[0046] R b Each is independently selected from halogen or C 1-3 alkyl;

[0047] R3 is selected from C 1-3 Alkyl, C 1-3 alkoxy, -CH2R4 or -CH2OR4, wherein the C 1-3 Alkyl and C 1-3 The alkoxy group may be optionally replaced by one, two, or three halogens;

[0048] R4 is selected from phenyl or 5-6 heteroaryl, wherein the phenyl and 5-6 heteroaryl are optionally independently substituted by 1, 2 or 3 Rs;

[0049] R is selected from halogen or C. 1-3 alkyl;

[0050] A1, A2, A3 and A4 are each independently selected from CH or N, and 0, 1 or 2 of A1, A2, A3 and A4 are selected from N, and the rest are selected from CH.

[0051] In some embodiments of the present invention, R1 is selected from F or methyl, and other variables are as defined in the present invention.

[0052] In some embodiments of the present invention, the two R1s and the atoms connected to them form C. 3-6 Cycloalkyl groups, and other variables as defined in this invention.

[0053] In some embodiments of the present invention, the two R1s and the atoms connected to them form a cyclopropyl group, and other variables are as defined in the present invention.

[0054] In some embodiments of the present invention, the above-mentioned structural unit Selected from Other variables are as defined in this invention.

[0055] In some embodiments of the present invention, R2 is selected from tert-butyl, and other variables are as defined in the present invention.

[0056] In some embodiments of the present invention, R3 is selected from -CF3, and other variables are as defined in the present invention.

[0057] The present invention also provides the compound shown in (I-1) or a pharmaceutically acceptable salt thereof.

[0058]

[0059] Wherein, R1 and n are as defined in this invention.

[0060] Some solutions in this invention are derived from arbitrary combinations of the above-mentioned variables.

[0061] This invention provides compounds of the following formula or pharmaceutically acceptable salts thereof:

[0062]

[0063]

[0064] The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the above-described compound or a pharmaceutically acceptable salt thereof.

[0065] The present invention also provides the use of the above-described compounds or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating coronavirus infections.

[0066] The present invention also provides the use of the above-described compounds or pharmaceutically acceptable salts thereof or the above-described compositions in the preparation of medicaments for treating coronavirus infection.

[0067] In some embodiments of the present invention, the coronavirus infection described above is selected from COVID-19.

[0068] Technical effect

[0069] The compounds of this invention exhibit good in vitro activity against the novel coronavirus Mpro protease; good in vitro anti-coronavirus activity at the cellular level; and are non-cytotoxic.

[0070] Definitions and Explanations

[0071] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0072] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0073] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound having specific substituents discovered in this invention with a relatively non-toxic acid or base. When the compounds of this invention contain relatively acidic functional groups, a base addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts or similar salts. When the compounds of this invention contain relatively basic functional groups, an acid addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds of this invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0074] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0075] Unless otherwise stated, the term "isomer" is intended to include geometric isomers, cis-trans isomers, stereo isomers, enantiomers, optical isomers, diastereomers and tautomers.

[0076] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0077] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0078] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0079] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0080] Unless otherwise stated, "(+)" indicates right-handed rotation, "(-)" indicates left-handed rotation, and "(±)" indicates racemic rotation.

[0081] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key Or straight dashed key

[0082] Unless otherwise specified, when a group has one or more connectable sites, any one or more of these sites can be connected to other groups by chemical bonds. The chemical bonds connecting these sites to other groups can be straight solid lines. Straight dashed key or wavy line For example, a straight solid line bond in -OCH3 indicates that the oxygen atom in that group is connected to other groups; The straight dashed bond in the diagram indicates that the group is connected to other groups through both ends of the nitrogen atom in the group; The wavy lines in the diagram indicate that the carbon atoms at positions 1 and 2 of the phenyl group are linked to other groups.

[0083] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0084] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0085] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated, and the auxiliary group is cleaved to provide the desired enantiomer in pure form. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0086] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0087] The terms “optional” or “optionally” refer to events or conditions that may occur but are not required to occur as described below, and the description includes both cases where said events or conditions occur and cases where said events or conditions do not occur.

[0088] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified, and the type and number of substituents can be arbitrary on a chemically feasible basis.

[0089] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0090] The term "therapeutic effective amount" means the amount of the compound of the present invention used to treat or prevent a particular disease, condition, or disorder; (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder; or (iii) to prevent or delay the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of the compound of the present invention constituting a "therapeutic effective amount" varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by those skilled in the art based on their own knowledge and the content of this disclosure.

[0091] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0092] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0093] When the number of a substituent is 0, it means that the substituent does not exist. For example, -A-(R)0 means that the structure is actually -A.

[0094] When a substituent is vacant, it means that the substituent does not exist. For example, if X is vacant in AX, it means that the structure is actually A.

[0095] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0096] When a substituent can be cross-bonded to two or more atoms on a ring, this substituent can bond with any atom on that ring, for example, structural units. This indicates that the substituent R can be substituted at any position on the cyclohexyl or cyclohexadiene. When the listed substituents do not specify which atom they are attached to the substituted group, such substituents can be bonded to any of their atoms. For example, a pyridyl group as a substituent can be attached to the substituted group through any carbon atom on the pyridine ring.

[0097] When the listed linking groups do not specify their linking direction, the linking direction is arbitrary, for example, The linker group L is -MW-. In this case, -MW- can connect ring A and ring A in the same direction as the reading order from left to right to form a ring. Alternatively, rings A and A' can be connected in the opposite direction to the left-to-right reading order to form a ring. The combination of linking groups, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0098] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of that group can be connected to other groups by chemical bonds. When the chemical bond connection is non-directional and the connectable site contains H atoms, the number of H atoms at that site will decrease accordingly with the number of chemical bonds connected, resulting in a group with a corresponding valence. The chemical bonds connecting the site to other groups can be straight solid line bonds. Straight dashed key or wavy line For example, a straight solid line bond in -OCH3 indicates that the oxygen atom in that group is connected to other groups; The straight dashed bond in the diagram indicates that the group is connected to other groups through both ends of the nitrogen atom in the group; The wavy lines in the diagram indicate that the phenyl group is connected to other groups through the carbon atoms at positions 1 and 2.

[0099] Unless otherwise specified, the term "C" 1-3 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C 1-3 Alkyl groups include C 1-2 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), etc.

[0100] Unless otherwise specified, the term "C" 1-3"Alkoxy" refers to alkyl groups containing 1 to 3 carbon atoms that are attached to the rest of the molecule by an oxygen atom. The C 1-3 Alkoxy groups include C 1-2 C 2-3 C3 and C2 alkoxy groups, etc. 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

[0101] Unless otherwise specified, the number of atoms in a ring is usually defined as the elemental number of the ring. For example, a “5-7 elemental ring” refers to a “ring” with 5-7 atoms arranged around it.

[0102] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 C 11 and C 12 It also includes any range from n to n+m, such as C 1-12 Including C 1-3 C 1-6 C 1-9 C 3-6 C 3-9 C 3-12 C 6-9 C 6-12 and C 9-12 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-12-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings, and also any range from n to n+m. For example, 3-12-membered rings include 3-6-membered, 3-9-membered, 5-6-membered, 5-7-membered, 6-7-membered, 6-8-membered, and 6-10-membered rings, etc.

[0103] Unless otherwise specified, "C 3-6 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, including monocyclic, bicyclic, and tricyclic systems, wherein bicyclic and tricyclic systems include spirocyclic, fused, and bridged rings. The C 3-6 Cycloalkyl groups include C 3-5 C 4-6 C 4-5 Or C 5-6 etc.; it can be monovalent, divalent, or polyvalent. C 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0104] Unless otherwise specified, "C 3-10 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 10 carbon atoms, including monocyclic, bicyclic, and tricyclic systems, wherein bicyclic and tricyclic systems include spirocyclic, fused, and bridged rings. The C 3-10 Cycloalkyl groups include C 3-8 C 3-6 C 3-5 C 4-10 C 4-8 C 4-6 C 4-5 C 5-8 Or C 5-6 etc.; it can be monovalent, divalent, or polyvalent. C 3-10 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, norbornyl, [2.2.2]bicyclooctane, etc.

[0105] Unless otherwise specified, the term "3-10 membered heterocyclic alkyl" on its own or in combination with other terms respectively refers to a saturated cyclic group consisting of 3 to 10 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, N, P, or Se, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen, sulfur, and phosphorus heteroatoms may optionally be oxidized (i.e., NO, S(O)). p and P(O) p (where p is 1 or 2). It includes monocyclic, bicyclic, and tricyclic systems, with bicyclic and tricyclic systems including spirocyclic, fused, and bridged rings. Furthermore, regarding the "3-10 membered heterocyclic alkyl," the heteroatom can occupy the connection position between the heterocyclic alkyl group and the rest of the molecule. The 3-10 membered heterocyclic alkyl groups include 3-8, 3-6, 3-5, 4-6, 5-6, 4, 5, and 6 membered heterocyclic alkyl groups, etc. Examples of 3-10 membered heterocyclic alkyl groups include, but are not limited to, azirrobutyl, oxacyclobutyl, thiocyclobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperidinyl and 2-piperidinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxane, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl, hexahydropyridazinyl, homopiperidinyl, homopiperidinyl, or dioxaneheptyl, etc.

[0106] Unless otherwise specified, the terms "5-6 membered heteroaryl" and "5-6 membered heteroaryl" are used interchangeably in this invention. The term "5-6 membered heteroaryl" refers to a monocyclic group with a conjugated π-electron system consisting of 5 to 6 ring atoms, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). p (where p is 1 or 2). The 5-6 membered heteroaryl group can be attached to the rest of the molecule via a heteroatom or a carbon atom. The 5-6 membered heteroaryl group includes both 5-membered and 6-membered heteroaryl groups. Examples of the 5-6 membered heteroaryl group include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), and triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl). (and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isooxazolyl, 4-isooxazolyl and 5-isooxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0107] Unless otherwise specified, the term “halogen” or “halogen” itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.

[0108] Unless otherwise stated, when two substituents and their attached atoms form a ring, the "attached atoms" can be the same atom or different atoms. For example, in this invention, "two R1s and their attached atoms form a C..." 3-6 "Cycloalkyl" can form spirocyclic, bridged, or fused rings with ring A.

[0109] The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0110] The structures of the compounds of this invention can be confirmed by conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, that absolute configuration can be confirmed by conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) is used, where the cultured single crystal is used to collect diffraction intensity data using a Bruker D8 venture diffractometer with CuKα radiation as the light source. The scanning method is as follows: After scanning and collecting relevant data, the crystal structure can be further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0111] The solvent used in this invention is commercially available.

[0112] The following abbreviations are used in this invention: HATU represents 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate; THF represents tetrahydrofuran; Boc represents tert-butyloxycarbonyl; n-BuLi represents n-butyllithium; HCl represents hydrochloric acid; ACN represents acetonitrile; MeOH represents methanol; EtOAc or EA represents ethyl acetate; TEA represents triethylamine; DIEA or DIPEA represents N,N-diisopropylethylamine; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; HPLC represents high performance liquid chromatography; TLC represents thin-layer chromatography; PEG represents polyethylene glycol.

[0113] Compounds are named according to conventional naming principles in the field or using Software naming conventions are used; commercially available compounds use supplier catalog names. Detailed Implementation

[0114] The present invention will be described in detail below with reference to embodiments, but this does not imply any adverse limitation on the invention. The present invention has been described in detail, and specific embodiments thereof have been disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope thereof.

[0115] Example 1

[0116]

[0117] Synthesis route:

[0118]

[0119] Step 1: Synthesis of Compounds 1-2

[0120] Compound 1-1 (5 g, 34.93 mmol) was dissolved in methanol (50 mL), and an aqueous sodium hydroxide solution (4 M, 43.66 mL) was added. The mixture was stirred at 20 °C for 4 h. Citric acid (50 mL), saturated brine (40 mL), and ethyl acetate (50 mL) were added to the reaction system for extraction. The organic phase was separated, dried over anhydrous sodium sulfate, and evaporated to dryness to obtain compound 1-2. 1 H NMR(400MHz, DMSO-d6)δ=–2.56-12.41(m,1H),–7.67-7.59(m,1H),–7.26-7.20(m,1H),–3.98-3.88(m,1 H),–3.34-3.30(m,1H),–3.19-3.08(m,2H),–2.36-2.08(m,2H),–1.69-1.50(m,2H),–1.41-1.32(m,9H).

[0121] Step 2: Synthesis of compounds 1-3

[0122] Compounds 1-2 (3.5 g, 12.95 mmol) were dissolved in N,N-dimethylformamide (40 mL), followed by the sequential addition of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (2.11 g, 11.01 mmol), 1-hydroxybenzotriazole (1.63 g, 12.04 mmol), and N,O-dimethylhydroxylamine hydrochloride (1.26 g, 12.95 mmol). The mixture was cooled to 0 °C, and triethylamine (1.22 g, 12.04 mmol, 1.68 mL) was slowly added dropwise. The mixture was then restored to 20 °C and reacted for 16 hours. The pH was adjusted to between 3 and 5 with saturated citric acid (50 mL), and ethyl acetate (80 mL) was added. The ethyl acetate was washed with saturated sodium carbonate solution (60 mL) and then with saturated brine (50 mL). The organic layer was dried and concentrated under reduced pressure to obtain the crude product. Compounds 1-3 were obtained by purification by silica gel column chromatography (dichloromethane:methanol = 1:0 to 20:1). 1 H NMR(400MHz, DMSO-d6)δ=–7.98-7.92(m,2H),–7.22-7.13(m,1H),–4.48-4.37(m,1H),–3.75-3.68(m,2H),–3.33- 3.30(m,1H),–3.19-3.12(m,2H),–3.12-3.08(m,1H),–2.75-2.71(m,5H),–2.52-2.47(m,1H),–1.45-1.30(m,9H).

[0123] Step 3: Synthesis of compounds 1-4

[0124] 2-Bromobenzothiazole (3.39 g, 15.85 mmol) was dissolved in THF (30 mL), purged three times with nitrogen, cooled to -78 °C, and a tetrahydrofuran solution of n-butyllithium (2.5 M, 5.07 mL) was slowly added dropwise. The mixture was stirred for 30 min, followed by the slow addition of a tetrahydrofuran solution of compounds 1-3 (0.5 g, 1.59 mmol) (5 mL). The reaction was allowed to proceed for 1 hour. A saturated ammonium chloride solution (40 mL) was added to the reaction mixture, and the mixture was stirred for 10 min. Ethyl acetate (50 mL) was added, and the mixture was extracted three times. The organic phases were combined, washed with saturated brine (50 mL), and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 1:0–20:1) to obtain compounds 1-4.

[0125] Step 4: Synthesis of compounds 1-5

[0126] Compounds 1-4 (30 mg, 77.03 μmol) were dissolved in hydrochloric acid / ethyl acetate solution (5 mL), stirred at 20 °C for 1 hr, and the reaction solution was directly concentrated under reduced pressure to obtain compounds 1-5.

[0127] MS–ESI m / z:[M+H] + =290.1.

[0128] Step 5: Synthesis of steps 1-7

[0129] Toluene (4 mL) and trimethylsilazomethane (2 M, 1.66 mL) were added to a methanol (2 mL) solution of starting materials 1-6 (400.00 mg, 1.66 mmol) at 0 °C, and the reaction was carried out at 20 °C for 16 h. The reaction solution was concentrated under reduced pressure to obtain the crude product. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 10:1) to obtain products 1-7. 1 H NMR(400MHz, CDCl3)δ=–4.39-4.20(m,1H),–3.87-3.68(m,4H),–2.74-2.62(m,1H),–1.98-1 .87(m,1H),–1.83-1.61(m,3H),–1.56-1.49(m,1H),–1.48-1.36(m,9H),–1.30-1.21(m,1H).

[0130] Step 6: Synthesis of steps 1-8

[0131] Hydrochloric acid / ethyl acetate (4M, 5mL) was added to the reaction flask containing reactants 1-7 (0.28g, 1.10mmol), and the mixture was reacted at 20°C for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the crude product. Compounds 1-8 were then obtained. 1H NMR(400MHz, CD3OD)δ=–4.21-4.07(m,2H),–3.94-3.79(m,3H),–3.03-2.91(m, 1H),–2.04-2.03(m,1H),–2.05-2.00(m,1H),–1.93-1.80(m,3H),1.73(s,2H).

[0132] Step 7: Synthesis of steps 1-9

[0133] At 0 °C, N-Boc-L-tert-leucine (0.21 g, 1.10 mmol), N,N-diisopropylethylamine (426.49 mg, 3.30 mmol, 574.79 μL), and HATU (627.38 mg, 1.65 mmol) were added to a solution of starting materials 1-8 (305.30 mg, 1.32 mmol) in 2 mL of N,N-dimethylformamide. The reaction mixture was reacted at 20 °C for 16 h. The reaction solution was then separated into 5% citric acid solution. The aqueous phase was extracted with ethyl acetate (20 mL × 2), and the organic phase was dried over anhydrous sodium sulfate. The mixture was filtered and concentrated under reduced pressure to obtain the crude product. The crude product was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 3:1) to give compounds 1-9. 1 H NMR (400MHz, CDCl3) δ = 5.21 (br d,J=9.6Hz,1H),–4.48-4.37(m,1H),–4.30-4.21(m,1H),–4.01-3.95(m,1H),–3.68-3.60(m,3H) ,–2.73-2.61(m,1H),–1.97-1.87(m,1H),–1.79-1.56(m,4H),–1.39-1.33(m,10H),0.97(s,9H).

[0134] Step 8: Synthesis of 1-10

[0135] Lithium hydroxide monohydrate (51.25 mg, 1.22 mmol) was added to a solution of starting materials 1-9 (0.3 g, 814.19 μmol) in tetrahydrofuran (2 mL) and water (1 mL), and the reaction was carried out at 20 °C for 16 h. 20 mL of 5% citric acid aqueous solution was added to the reaction mixture, followed by 20 mL of ethyl acetate. The mixture was separated, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Compounds 1-10 were then obtained. 1H NMR(400MHz, CDCl3)δ=5.24(d,J=9.9Hz,1H),–4.51-4.45(m,1H),–4.41-4.33( m,1H),–4.18-4.15(m,1H),–3.06-2.99(m,1H),–1.99-1.89(m,1H),–1.85-1.75 (m,3H),–1.59-1.49(m,2H),–1.46-1.42(m,9H),–1.05-1.01(m,9H).

[0136] Step 9: Synthesis of compounds 1-11

[0137] Compound 1-10 (0.5 g, 1.41 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (1.5 mL) was added. The reaction was carried out at 20 °C for 1 h. The reaction was then directly concentrated to give compound 1-11.

[0138] MS–ESI m / z:[M+H] + =255.3.

[0139] Step 10: Synthesis of compounds 1-12

[0140] Compound 1-11 (0.45 g, 1.55 mmol) was dissolved in methanol (10 mL). Methyl trifluoroacetate (1.98 g, 15.48 mmol) was added to the reaction system, followed by triethylamine (939.57 mg, 9.29 mmol). The reaction was stirred at 38 °C for 12 h. The crude reaction solution was directly concentrated under reduced pressure. The crude product was dissolved in dichloromethane (30 mL) and washed with 1 N hydrochloric acid solution (10 mL). The organic phase was washed with saturated brine (20 mL), filtered, and concentrated to obtain compound 1-12.

[0141] MS–ESI m / z:[M+H] + =351.1.

[0142] Step 11: Synthesis of Compound 1

[0143] Compounds 1-12 (25.1 mg, 77.04 μmol) and 1-5 (29.69 mg, 84.74 μmol) were dissolved in acetonitrile (5 mL). The reaction mixture was cooled to 0 °C, and 1-ethyl-(3-dimethylaminopropyl)carbamate dimethylimine hydrochloride (16.25 mg, 84.74 μmol) was added, followed by dropwise addition of pyridine (24.37 mg, 308.15 μmol). The reaction mixture was carried out at 0 °C for 2 hours. The reaction mixture was extracted with ethyl acetate (10 mL) and 1 M hydrochloric acid (5 mL). The organic layer was washed with saturated sodium bicarbonate aqueous solution (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 1 was purified by preparative TLC (ethyl acetate, Rf = 0.5).

[0144] MS–ESI m / z:[M+H] + =621.67. 1 H NMR (400MHz, CDCl3) δppm–8.16-8.21(m,1H),–7.95-8.03(m,1H),–7.82-7.87(m, 1H),–7.52-7.62(m,2H),–7.00-7.12(m,2H),–5.77-5.91(m,1H),–5.41-5.51(m,1 H),4.67(s,1H),–4.41-4.54(m,1H),4.03(s,1H),–3.31-3.44(m,2H),–2.87-2.96 (m,1H),–2.54-2.73(m,2H),–1.74-2.32(m,7H),–1.23-1.35(m,1H),1.10(s,8H).

[0145] Example 2

[0146]

[0147]

[0148] Step 1: Synthesis of Compound 2-2

[0149] Compound 2-1 (5 g, 54.32 mmol) was dissolved in methanol (50 mL) and refluxed at 70 °C for 48 h. The reaction system was concentrated under reduced pressure to obtain the crude target product. The crude product had high purity and was directly used in the next reaction to obtain compound 2-2. 1 H NMR (400MHz, CDCl3) δ = 4.81 (s, 1H), 3.77 (s, 3H), 3.43 (s, 3H).

[0150] Step 2: Synthesis of compounds 2-3

[0151] Compound 2-2 was dissolved in toluene (3 mL), cooled to 0 °C, and compound (R)-(+)-phenylethylamine (1.5 g, 12.38 mmol, 1.60 mL) was slowly added dropwise. The mixture was stirred at 20 °C for 1 h. Ethyl acetate (60 mL) and saturated brine (30 mL) were added to the reaction system for extraction to obtain the organic phase. The extract was dried over anhydrous sodium sulfate and evaporated to dryness to obtain the crude product. Purification was performed by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0–5:1) to obtain the target compound 2-3. 1 H NMR (400MHz, CDCl3) δ = –7.95-7.56(m,1H), –7.31-7.17(m,5H), –4.71-4.40(m,1H), –3.95-3.71(m,3H), –1.67-1.51(m,3H).

[0152] Step 3: Synthesis of compounds 2-4

[0153] Compound 2-3 (0.5 g, 2.61 mmol) was dissolved in 2,2,2-trifluoroethanol (5 mL), and trifluoroacetic acid (313.04 mg, 2.75 mmol, 203.28 μL) was added. The mixture was cooled to -10 °C and stirred for 1 h. Cyclopentadiene (207.40 mg, 3.14 mmol) was slowly added dropwise while maintaining the temperature at -10 °C, and stirring was continued for 0.5 h. The reaction mixture was concentrated under reduced pressure, and methyl tert-butyl ether (60 mL) and saturated sodium bicarbonate solution (30 mL × 2) were added. The mixture was stirred for 10 min, extracted, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The solution was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0–5:1) to obtain compound 2-4, the configuration of which was confirmed by two-dimensional NMR. 1 HNMR(400 MHz, CDCl3)δ=–7.34-7.18(m,5H),–6.59-6.41(m,1H),6.31(dd,J=1.6,5.6Hz,1H),4.35(br d,J=1.3Hz,1H),3.39(s,3H),–3.18-3.03(m,1H),2.95(br s,1H),–2.33-2.22(m,1H),2.14(br d,J=8.4Hz,1H),–1.54-1.41(m,4H).

[0154] MS–ESI m / z:[M+H] + =258.2.

[0155] Step 4: Synthesis of compounds 2-5

[0156] Compounds 2-4 (100.00 mg, 388.61 μmol) were dissolved in tetrahydrofuran (1.25 mL), cooled to -70 °C, and a borane tetrahydrofuran complex (1 M, 427.47 μL) was slowly added dropwise. The mixture was then slowly heated to 20 °C and stirred for 1 h. The mixture was cooled to 0 °C, and a 10% sodium hydroxide aqueous solution (0.55 mL) and a 30% hydrogen peroxide solution (220.28 mg, 1.94 mmol, 186.68 μL) were added. The mixture was then slowly heated to 20 °C and stirred for 1 h. A saturated sodium thiosulfate solution (10 mL) was added to the reaction system, and the reaction was quenched by stirring for 10 min. The mixture was then extracted with saturated brine (20 mL) and ethyl acetate (60 mL × 2), and the organic phase was separated. A small amount of the sample solution was taken, and the pH was adjusted to less than 8 with 3% citric acid. After testing with starch-potassium iodide test paper and finding negative, the solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure at 30 °C. Compounds 2-5 were obtained by purification by silica gel column chromatography (petroleum ether: ethyl acetate = 1:0 to 5:1). 1 H NMR (400MHz, CDCl3) δ = –7.30-7.13 (m, 5H), 3.93 (br d, J = 6.5Hz, 1H), 3.78 (br s,1H),–3.70-3.54(m,1H),–3.39-3.32(m,1H),–3.31-3.24(m,3H),–2.49-2.40(m,1H),2.26(s,1H),–2.09-2.00(m,1H),1.72(br d,J=10.1Hz,1H),1.46(br d,J=6.5Hz,1H),–1.41-1.33(m,3H).

[0157] MS–ESI m / z:[M+H] + =276.1.

[0158] Step 5: Synthesis of hydrochloride salts of compounds 2-6

[0159] Compound 2-5 (3 g, 10.90 mmol) was dissolved in ethanol (80 mL), and concentrated hydrochloric acid (1.19 g, 32.69 mmol) and wet palladium on carbon (15 g, 10.68 mmol, 5%) were added. The reaction was stirred at 20 °C for 16 h. The reaction solution was filtered through diatomaceous earth and then directly evaporated to dryness to give the crude hydrochloride of compound 2-6.

[0160] MS–ESI m / z:[M+H] + =172.0.

[0161] Step 6: Synthesis of compounds 2-8

[0162] Compound 2-7 (1.87 g, 10.90 mmol) was dissolved in N,N-dimethylformamide (20 mL), and HATU (4.78 g, 12.58 mmol) and diisopropylethylamine (4.34 g, 33.55 mmol) were added. After stirring for 30 min, the hydrochloride salt of compound 2-6 (190 mg, 1.12 mmol) was added. The reaction mixture was stirred at 20 °C for 16 h. Water (15 mL) was added to the reaction mixture, and the mixture was extracted twice with ethyl acetate (60 mL). The organic phases were combined, washed twice with 5% citric acid (30 mL), washed four times with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Compound 2-8 was purified by column chromatography (petroleum ether:ethyl acetate = 3:1). 1 H NMR (400MHz, CDCl3) δ=–5.28-5.16(m,1H),4.50(br s,1H),4.28(d,J=9.8Hz,1H),3.92(s,1H),3.74(s,3H),2.81(s,1H),2.67(s,1H),2.17(br dd,J=6.1,12.7Hz,1H),–1.99-1.93(m,1H),–1.90-1.84(m,1H),1.59(br d,J=13.3Hz,2H),1.43(s,9H),1.04(s,9H).

[0163] MS–ESI m / z:[M+H] + =385.2.

[0164] Step 7: Synthesis of compounds 2-9

[0165] Compounds 2-8 (500 mg, 1.30 mmol) were dissolved in acetonitrile (7.5 mL), and 2-iodobenzoic acid (976.31 mg, 3.49 mmol) was added. The mixture was stirred at 60 °C for 16 h. The reaction solution was filtered directly through diatomaceous earth and evaporated to dryness. Compounds 2-9 were obtained without purification.

[0166] MS–ESI m / z:[M-55] + =327.1.

[0167] Step 8: Synthesis of Compounds 2-10

[0168] Compound 2-9 (480 mg, 1.26 mmol) was dissolved in dichloromethane (4.8 mL), cooled to 0 °C, and diethylaminotrifluoride (1.01 g, 6.28 mmol) was added. The mixture was stirred at 20 °C for 16 h. The reaction solution was then slowly added to a saturated sodium bicarbonate solution (20 mL) at 0 °C, and extracted twice with dichloromethane (20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 2-10 was obtained without purification.

[0169] MS–ESI m / z:[M-100] + =304.0.

[0170] Step 9: Synthesis of Compound 2-11

[0171] Compound 2-10 (475 mg, 1.17 mmol) was dissolved in tetrahydrofuran (5.5 mL), water (1.84 mL), and methanol (1.84 mL). Lithium hydroxide monohydrate (147.84 mg, 3.52 mmol) was added, and the mixture was stirred at 20 °C for 16 h. Water (20 mL) and 5% citric acid (25 mL) were added to the reaction mixture, and the mixture was extracted twice with ethyl acetate (40 mL). The combined organic phases were washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Compound 2-11 was obtained without purification. 1 H NMR (400MHz, CDCl3) δ = 5.18 (br d,J=9.8Hz,1H),–4.65-4.53(m,2H),4.31(d,J=9.9Hz,1H),3.33(brd,J=7. 3Hz,1H),–2.55-2.41(m,1H),–2.33-2.07(m,3H),1.45(s,9H),1.03(s,9H).

[0172] MS–ESI m / z:[M-55] + =335.1.

[0173] Step 10: Synthesis of Compounds 2-12

[0174] Compounds 2-10 (160 mg, 409.81 μmol) and 1-5 (160.23 mg, 491.78 μmol) were dissolved in acetonitrile (2 mL), cooled to 0 °C, and 1-ethyl-(3-dimethylaminopropyl)carbamate dimethylimine hydrochloride (79.35 mg, 413.91 μmol) was added. Then, pyridine (129.66 mg, 1.64 mmol) was added dropwise, and the reaction was stirred at 0 °C for 2 h. Water (15 mL) was added to the reaction solution, and the mixture was extracted twice with ethyl acetate (30 mL). The organic phases were combined, washed twice with 5% citric acid (20 mL), washed once with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Compound 2-12 was purified by column chromatography (dichloromethane:methanol = 20:1). 1 H NMR (400MHz, CDCl3) δ=8.19(s,1H),–8.02-7.97(m,1H),–7.61-7.55(m,2H),5.83(q,J=7.8Hz,1H),5.55(s,1H),5.26(br d,J=10.4Hz,1H),–4.58-4.51(m,2H),4.31(d,J=10.1Hz,1H),–3.40-3.33(m,2H),3.20(br d,J=6.6Hz,1H),–2.67-2.53(m,2H),–2.36-2.16(m,6H),–2.13-2.07(m,1H),1.94(br d,J=10.0Hz,1H),1.45(s,9H),1.05(s,9H).

[0175] MS–ESI m / z:[M+H] + =662.3.

[0176] Step 11: Synthesis of the trifluoroacetate of compound 2-13

[0177] Compound 2-12 (210 mg, 317.34 μmol) was dissolved in dichloromethane (2.1 mL) and trifluoroacetic acid (0.7 mL), and the reaction was stirred at 20 °C for 2 h. The reaction mixture was evaporated to dryness using an oil pump, and the dichloromethane was repeatedly evaporated until a pale yellow foam was formed. The trifluoroacetate salt of compound 2-13 was obtained.

[0178] MS–ESI m / z:[M+H] + =562.2.

[0179] Step 12: Synthesis of Compound 2

[0180] The trifluoroacetate salt of compound 2-13 (170 mg, 302.68 μmol) was dissolved in methanol (3.4 mL), followed by the addition of triethylamine (245.03 mg, 2.42 mmol) and methyl trifluoroacetate (387.59 mg, 3.03 mmol). The reaction mixture was heated to room temperature (38 °C) and stirred for 16 h. The reaction solution was directly evaporated to dryness, dissolved in water (10 mL) and ethyl acetate (10 mL), and the solution was adjusted to acidity with 5% citric acid (10 mL). The mixture was separated, and extracted twice with ethyl acetate (10 mL). The combined organic phases were washed twice with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The solution was then analyzed by preparative TLC (ethyl acetate, R...). f =0.5) was purified to obtain compound 2. 1 H NMR (400MHz, CDCl3) δ = –8.22-8.11(m,1H), –8.01-7.95(m,1H), –7.61-7.52(m ,2H),4.68(d,J=9.2Hz,1H),–4.61-4.53(m,2H),–3.44-3.35(m,2H),3.20(br d,J=5.7Hz,1H),–2.81-2.73(m,1H),–2.33-1.88(m,9H),1.07(s,9H).

[0181] MS–ESI m / z:[M+H] + =658.3.

[0182] Example 3

[0183]

[0184] Synthesis route:

[0185]

[0186] Step 1: Synthesis of compound 3-3

[0187] Compound 3-1 (4.62 g, 21.01 mmol) was dissolved in N,N-dimethylformamide (50 mL), and HATU (9.22 g, 24.24 mmol) and N,N-diisopropylethylamine (8.35 g, 64.64 mmol, 11.26 mL) were added. The mixture was stirred for 0.5 hours, and then compound 3-2 (5 g, 16.16 mmol) was added. The mixture was stirred at 20 °C for 1 hour. The reaction mixture was extracted with water (150 mL) and ethyl acetate (30 mL × 2). The combined organic phases were washed with 5% citric acid (10 mL) and brine (20 mL × 4), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Compound 3-3 was purified by column chromatography (petroleum ether:ethyl acetate = 5:1).

[0188] MS–ESI m / z:[M+H] + =475.6.

[0189] Step 2: Synthesis of compounds 3-4

[0190] Compound 3-3 (6 g, 12.64 mmol) and solvents tetrahydrofuran (90 mL), methanol (30 mL), and water (30 mL) were added to a dry single-necked flask. Lithium hydroxide monohydrate (1.59 g, 37.92 mmol) was then added, and the reaction was carried out at 25 °C for 12 hours. In a conical flask, 1 M hydrochloric acid was slowly added dropwise to adjust the pH to 7. Then, 20 mL of water and 20 mL of dichloromethane were added, and the mixture was extracted three times. The organic phase was separated, washed with 20 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the residue. Compound 3-4 was then obtained.

[0191] MS–ESI m / z:[M+H] + =447.5.

[0192] Step 3: Synthesis of compounds 3-5

[0193] Compounds 3-4 (0.35 g, 783.74 μmol) and 1-5 (0.2 g, 691.20 μmol) were dissolved in acetonitrile (10 mL), cooled to 0 °C, and 1-(3-dimethylaminopropyl)-3-acetaldehyde hydrochloride (150.24 mg, 783.74 μmol) was added. Pyridine (225.43 mg, 2.85 mmol, 230.03 μL) was added dropwise, and the reaction was carried out at 0 °C for 2 hours. Water (15 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 2). The combined organic phases were washed with 5% citric acid (10 mL) and brine (20 mL × 4), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Compound 3-5 was purified by column chromatography (dichloromethane:methanol = 5:1).

[0194] MS–ESI m / z:[M+H] + =718.9.

[0195] Step 4: Synthesis of compound 3-6 trifluoroacetate

[0196] Compound 3-5 (0.3 g, 417.88 μmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (2.5 mL), and stirred at 15 °C for 1 hour. The reaction solution was concentrated under reduced pressure to obtain trifluoroacetate of compound 3-6. The reaction solution was directly added to the next step without purification.

[0197] MS–ESI m / z:[M+H] + =618.8.

[0198] Step 5: Synthesis of Compound 3

[0199] Compound 3-6 trifluoroacetate (0.17 g, 275.17 μmol) was dissolved in dichloromethane (1 mL), cooled to 0 °C, and pyridine (152.36 mg, 1.93 mmol, 155.47 μL) and trifluoroacetic anhydride (231.18 mg, 1.10 mmol, 153.10 μL) were added. The reaction was carried out at 0 °C for 0.5 h. Water (15 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with 5% citric acid (10 mL) and brine (20 mL × 4), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. The crude product was separated by preparative HPLC (column type: Waters Xbridge Prep OBD C18 150*40 mm*10 μm; mobile phase: [water (NH4HCO3)-acetonitrile]; acetonitrile %: 50%-80%, 8 min). The fraction was concentrated under reduced pressure to obtain compound 3.

[0200] MS–ESI m / z:[M+H] + =714.8. 1 H NMR (400MHz, CDCl3) δ=8.25-8.14(m,1H),8.05-7.94(m,1H),7.63-7.51(m,2H),5.87-5.76(m,1H),5.76-5.75(m,1H),5.71-5. 59(m,1H),4.63-4.45(m,1H),4.41-4.19(m,1H),3.99-3.75(m,1H),3.65-3.32(m,3H),2.72-1.95(m,10H),1.75-1.37(m,16H).

[0201] Example 4

[0202]

[0203] Synthesis route:

[0204]

[0205] Step 1: Synthesis of compound 4-2

[0206] In a pre-dried three-necked flask, 10 g (34.93 mmol) of starting compound 1-1 was added, followed by 100 mL of methanol as solvent. The mixture was purged with nitrogen three times. The reactor was placed in an ice bath, and the system temperature was maintained at -10 to -5 °C. Sodium borohydride (5.39 g, 142.47 mmol) was slowly added dropwise. After the addition was complete, the mixture was stirred at 15 °C for 12 hours. Water (100 mL) was added to the reaction mixture, and the mixture was stirred for 10 minutes. The mixture was extracted with ethyl acetate (200 mL × 4). The organic layer was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated under reduced pressure to obtain compound 4-2.

[0207] MS–ESI m / z:[M+H] + =259.2.

[0208] Step 2: Synthesis of Compound 4-3

[0209] Compound 4-2 (8 g, 30.97 mmol) and dichloromethane (80 mL) were added to a dry three-necked flask at 15 °C, and stirring was started. Subsequently, sodium bicarbonate (2.73 g, 32.52 mmol, 1.26 mL) and Desmond-Martin periodide (13.79 g, 32.52 mmol) were added. The reaction solution changed from turbid to clear. After purging with nitrogen three times and stirring for approximately 20 min, the reaction solution became a white suspension. The mixture was stirred at 15 °C for 16 h. The reaction solution was quenched with sodium thiosulfate (100 mL), extracted with dichloromethane (100 mL × 3), and the organic phases were combined. The organic phase was washed with a saturated sodium bicarbonate solution (200 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was separated by an automated column chromatography (gradient elution: dichloromethane: methanol = 100:0-90:10) to obtain compound 4-3.

[0210] Step 3: Synthesis of compounds 4-5

[0211] Compound 4-4 (111 mg, 931.83 μmol) was added to a pre-dried three-necked flask, and the solvent tetrahydrofuran (1 mL) was added. Nitrogen gas was then introduced three times, and the reactor was placed in an ice bath. The system temperature was maintained at 0–5 °C. A tetrahydrofuran solution of the isopropyl magnesium lithium chloride complex (2 M, 465.91 μL) was slowly added dropwise. After the addition was complete, the mixture was stirred at 0 °C for 0.5 h. Subsequently, while maintaining the temperature at 0–5 °C, a mixed solvent of compound 4-3 (0.3 g, 1.17 mmol) and tetrahydrofuran (3 mL) was added to the above system. After the addition was complete, the temperature was slowly raised to 15 °C, and stirring was continued for 2 h. The reaction was quenched by adding 5 mL of saturated ammonium chloride aqueous solution, and the mixture was extracted with ethyl acetate (10 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product was separated by an automated column chromatography (gradient elution: dichloromethane: methanol = 100:0–95:5) to obtain compound 4-5.

[0212] MS–ESI m / z:[M+1] + =376.1.

[0213] Step 4: Synthesis of hydrochloride salts of compounds 4-6

[0214] Substrate compounds 4-5 (0.2 g, 532.74 μmol) were dissolved in hydrochloric acid / ethyl acetate (4 M, 10.00 mL) and stirred for 1 h. The solution was then directly concentrated to give the hydrochloride salt of compound 4-6.

[0215] Step 5: Synthesis of compounds 4-7

[0216] Substrate compounds 3-4 (0.35 g, 783.74 μmol) were dissolved in N,N-dimethylformamide (10 mL), and HATU (447.00 mg, 1.18 mmol) was added. The mixture was stirred for 0.5 h. The hydrochloride salt of compounds 4-6 (258.92 mg) and diisopropylethylamine (405.17 mg, 3.13 mmol) were added separately to the reaction mixture, and the mixture was stirred at 20 °C for 1 h. The reaction mixture was extracted twice with water (150 mL) and ethyl acetate (30 mL). The combined organic phases were washed with 5% citric acid (10 mL) and brine (20 mL × 4), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. Compounds 4-7 were purified by TLC (petroleum ether:ethyl acetate = 3:1, Rf = 0.3) followed by column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain compounds 4-7.

[0217] MS–ESI m / z:[M+H] + =704.4.

[0218] Step 6: Synthesis of compounds 4-8

[0219] Compounds 4-7 (0.2 g, 284.15 μmol) were dissolved in dichloromethane (2 mL), and Dys-Martin periodide (361.55 mg, 852.44 μmol) was added to the reaction system. The mixture was stirred at 15 °C for 1 h. The mixture was washed with saturated sodium thiosulfate solution (10 mL) and saturated sodium bicarbonate solution (10 mL), extracted three times with dichloromethane (10 mL), and the organic phases were combined and washed with saturated brine (30 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compounds 4-8.

[0220] MS–ESI m / z:[M+H] + =702.3.

[0221] Step 7: Synthesis of trifluoroacetate of compound 4-9

[0222] Compound 4-8 (0.09 g, 128.23 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (462.00 mg, 4.05 mmol, 300.00 μL) was added to the reaction system. The reaction was stirred at 15 °C for 2 h. The reaction was directly concentrated to give compound 4-9 trifluoroacetate.

[0223] Step 8: Synthesis of Compound 4

[0224] Substrate compound 4-9 trifluoroacetate (0.034 g, 56.50 μmol) was dissolved in dichloromethane (1 mL), cooled to 0 °C, and pyridine (31.29 mg, 395.52 μmol) and trifluoroacetic anhydride (47.47 mg, 226.01 μmol) were added. The mixture was stirred for 0.2 h. The reaction mixture was washed with saturated sodium bicarbonate solution (10 mL), extracted three times with dichloromethane (10 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Preparative HPLC separation [Column type: Waters Xbridge BEH C18 100*30 mm*10 μm; Mobile phase: [H2O(NH4HCO3)-acetonitrile]; Acetonitrile %: 55%-75%, 8 min] yielded compound 4.

[0225] MS–ESI m / z:[M+H] + =698.3.

[0226] Example 5

[0227]

[0228] Synthesis route:

[0229]

[0230] Step 1: Synthesis of Compound 5-4

[0231] Thiazole[4,5-c]pyridine (2.16 g, 15.86 mmol) was dissolved in THF (15 mL), and the mixture was cooled to -78 °C under nitrogen protection. Then, n-BuLi (2.5 M THF solution, 6.35 mL) was added dropwise, and the mixture was stirred for 30 minutes. Next, a THF solution of compound 5-3 (1 g, 3.17 mmol) in 5 mL was added dropwise to the reaction mixture, and the mixture was stirred at -78 °C for 2 hours. The mixture was then slowly heated to 0 °C and stirred for 0.5 hours. After the reaction was complete, saturated ammonium chloride aqueous solution (20 mL) and water (20 mL) were added sequentially to quench the reaction. The mixture was extracted with ethyl acetate (80 mL * 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (DCM:MeOH = 1:0 to 20:1) and then preparatively separated by high performance liquid chromatography (neutral: column type: Ultimate C18 150*40mm*5μm; mobile phase: [H2O(NH4OH v / v)-ACN]; ACN%: 21%-51%, 20min) to obtain compounds 5-4.

[0232] MS–ESI m / z:[M+H] + =391.1.

[0233] Step 2: Synthesis of the hydrochloride salt of compound 5-5

[0234] Compound 5-4 (100 mg, 256.11 μmol) was dissolved in EtOAc (2 mL), and HCl / EtOAc (4 M, 6 mL) was added dropwise at 20 °C. After the addition was complete, the mixture was stirred at 20 °C for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (20 mL). No further purification was required. Concentration under reduced pressure yielded the hydrochloride salt of compound 5-5.

[0235] MS–ESI m / z:[M+H] + =290.9.

[0236] Step 3: Synthesis of compounds 5-6

[0237] The hydrochloride salt of compound 5-5 (70 mg, 214.20 μmol), compounds 3-4 (96 mg, 214.97 μmol), HATU (126.00 mg, 331.38 μmol), and TEA (32.72 mg, 323.30 μmol, 45 μL) were added to DMF (1 mL), and the mixture was stirred at 25 °C for 12 hours. After the reaction was complete, the mixture was quenched by dilution with water (20 mL) and saturated sodium bicarbonate solution (10 mL) sequentially. The mixture was extracted with ethyl acetate (50 mL * 3), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 5-6. The residue did not require further purification and was used directly in the next step.

[0238] MS–ESI m / z:[M+H] + =719.1.

[0239] Step 4: Synthesis of compound 5-7 hydrochloride

[0240] The crude compound 5-6 (180 mg, 250.38 μmol) was dissolved in EtOAc (2 mL), cooled to 0 °C, and HCl / EtOAc (4 M, 62.60 μL) was added dropwise. After the addition was complete, the mixture was reacted at 20 °C for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure to obtain compound 5-7 hydrochloride. MS–ESI m / z: 619.1 [M+H] + .

[0241] Step 5: Synthesis of Compound 5

[0242] The crude compound 5-7 hydrochloride (200 mg, 323.21 μmol) was dissolved in MeOH (10 mL), and TEA (163.57 mg, 1.62 mmol, 225 μL) and methyl trifluoroacetate (414.02 mg, 3.23 mmol, 326 μL) were added. The mixture was stirred at 38 °C for 12 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The residue was separated by high performance liquid chromatography (column type: Welch Xtimate C18 100*40 mm*3 μm; mobile phase: [H2O(HCl)-ACN]; ACN%: 30%-70%, 9.5 min) to obtain compound 5.

[0243] MS–ESI m / z:[M+H] + =715.1.

[0244] 1 H NMR(400MHz,CD3OD)δppm 9.80-9.53(m,1H),8.98-8.65(m,2H),5.85-5.49(m,1H),4.74-4.47(m,1H), 4.46-3.58(m,4H),3.44-3.35(m,1H),2.87-2.04(m,6H),2.03-1.24(m,22H).

[0245] Example 6

[0246]

[0247] Synthesis route:

[0248]

[0249] Step 1: Synthesis of Compound 6-2

[0250] 2-Bromothiazol[5,4-b]pyridine (340.98 mg, 1.59 mmol) was dissolved in THF (5 mL), and under nitrogen protection, the mixture was cooled to -78 °C. n-BuLi (2.5 M THF solution, 634.18 μL) was added dropwise, and the reaction was stirred for 30 minutes. Then, a THF solution of compound 1-3 (100 mg, 317.09 μmol) was added dropwise to the reaction mixture, and the mixture was stirred at -78 °C for 2 hours. After the reaction was complete, the mixture was quenched with water (20 mL) (internal temperature below -50 °C), extracted with ethyl acetate (30 mL * 3), and the organic phases were combined and concentrated under reduced pressure. The residue was purified by preparative thin-layer chromatography (EtOAc) to obtain compound 6-2.

[0251] MS–ESI m / z:[M+H] + =391.0.

[0252] 1 ¹H NMR (400MHz, CDCl₃) δppm 8.69 (dd, J = 1.5, 4.5Hz, 1H), 8.36 (dd, J = 1.3, 8.3Hz, 1H), 7.48 (dd, J = 4.8, 8.3Hz, 1H), 5.93–5.70 (m, 2H), 5.59–5.43 (m, 1H), 3.38–3.27 (m, 2H), 2.64–2.49 (m, 2H), 2.18–1.90 (m, 3H), 1.37 (s, 9H). Step 2: Synthesis of the hydrochloride salt of compound 6-3.

[0253] Compound 6-2 (0.1 g, 256.11 μmol) was dissolved in EtOAc (1 mL), and HCl / EtOAc (4 M, 64.03 μL) was added. The mixture was stirred at 20 °C for 3 hours. After the reaction was complete, the reaction solution was concentrated to remove the solvent, yielding the hydrochloride salt of compound 6-3, which was used directly in the next step.

[0254] MS-ESI m / z:[M+H] + =291.0.

[0255] Step 3: Synthesis of Compound 6-4

[0256] The hydrochloride salt of compound 6-3 (0.09 g, 309.98 μmol) and compound 3-4 (166.12 mg, 371.98 μmol) were dissolved in DMF (3 mL). HATU (235.73 mg, 619.96 μmol) and DIEA (200.31 mg, 1.55 mmol, 269.97 μL) were added, and the mixture was stirred at 20 °C for 1 hour. After the reaction was complete, EA (20 mL) and water (20 mL) were added to the reaction solution. The mixture was separated, and the aqueous phase was extracted twice with EA (20 mL). The combined organic phases were washed with saturated NaHCO3 (20 mL) and saturated NaCl (20 mL), and the organic phase was concentrated and evaporated to dryness. Compound 6-4 was obtained by column chromatography (DCM:MeOH = 1:0 to 20:1).

[0257] ESI m / z: [M+H] + =719.3.

[0258] Step 4: Synthesis of the hydrochloride salt of compound 6-5

[0259] Compound 6-4 (0.12 g, 166.92 μmol) was dissolved in EtOAc (1.3 mL), and HCl / EtOAc (4 M, 1.23 mL) was added. The mixture was stirred at 20 °C for 3 hours. After the reaction was complete, the reaction solution was concentrated to remove the solvent, yielding the hydrochloride salt of compound 6-5, which could be used directly in the next step without purification.

[0260] MS-ESI m / z:[M+H] + =619.3.

[0261] Step 5: Synthesis of Compound 6

[0262] Compound 6-5 (0.1 g, 161.61 μmol) was dissolved in MeOH (5 mL), and TEA (81.76 mg, 808.03 μmol, 112.47 μL) and methyl trifluoroacetate (206.94 mg, 1.62 mmol, 162.94 μL) were added. The mixture was stirred at 38 °C for 16 h. It was then dissolved in acetonitrile. The reaction solution was purified by prep-HPLC (column: Phenomenex C18 80*40 mm*3 μm; mobile phase: [H2O(NH3H2O)-ACN]; ACN%: 50%-80%, 8 min) to obtain compound 6. MS-ESI m / z: 715.2 [M+H] + . 1H NMR (400MHz, CDCl3) δppm 8.69-8.83(m,1H)8.41-8.53(m,1H)7.51-7.64(m,1H)5.56-5.92(m,2H)4.45-4.62(m,1H)4.20-4.43(m,1H)3.75-4.04(m,1H)3.2 8-3.69(m,3H)3.07-3.19(m,2H)2.70-2.99(m,3H)2.48-2.68(m,1H)2.17-2.28(m,1H)1.96-2.09(m,5H)1.69-1.72(m,1H)1.67(br d,J=16.31Hz,16H)1.40-1.49(m,3H).

[0263] Example 7

[0264]

[0265] Synthesis route:

[0266]

[0267] Step 1: Synthesis of Compound 7-3

[0268] A 50 mL three-necked flask was used. 7-2 (100 mg, 271.40 μmol) and 1-5 (78.53 mg, 271.40 μmol) were dissolved in 10 mL of DMF. Then, HATU (206.39 mg, 542.79 μmol) and DIEA (105.23 mg, 814.19 μmol, 141.82 μL) were added, and the mixture was stirred at 20 °C for 12 hours. After the reaction was complete, 30 mL of water was added to the resulting reaction solution, and the mixture was extracted twice with 30 mL of ethyl acetate. The organic phase was retained, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by vacuum distillation to obtain the crude product 7-3.

[0269] MS–ESI m / z:[M+Na] + =662.1.

[0270] Step 2: Synthesis of compound 7-4 hydrochloride

[0271] A 50 mL three-necked flask was used to dissolve 7-3 (158 mg, 246.95 μmol) in HCl / EtOAc (4 M, 5 mL), and the mixture was stirred at 20 °C for 12 hours. After the reaction was complete, the product 7-4 hydrochloride was obtained by vacuum distillation.

[0272] MS–ESI m / z:[M+H] + =540.0.

[0273] Step 3: Synthesis of Compound 7

[0274] 7-4 hydrochloride (160 mg, 296.47 μmol) was dissolved in 10 mL of MeOH in a 50 mL three-necked flask. Then, methyl trifluoroacetate (379.63 mg, 2.96 mmol, 298.92 μL) and TEA (180.00 mg, 1.78 mmol, 247.59 μL) were added, and the mixture was stirred at 20 °C for 12 hours. After the reaction was complete, the reaction solution was evaporated to dryness to obtain the crude product. The crude product was then separated by high-performance liquid chromatography (HPLC) (column: Ultimate C18 150*40 mm*5 μm; mobile phase: [water (HCl)-ACN]; ACN%: 40%-70%, 10 min) to obtain compound 7.

[0275] MS–ESI m / z:[M+H] + =636.1.

[0276] 1 H NMR (400MHz, CD3OD) δppm 8.24-8.18(m,1H)8.16-8.09(m,1H)7.71-7.56(m,2H)5.78(dd,J=11.8,3.3Hz,1H)4.72(s,1H)4.51(br s,1H)4.39(s,1H)3.41-3.33(m,2H)2.90-2.78(m,1H)2.59-2.47(m,2H)2.30-2.14(m,3H)2.14–1.91(m,4H)1.54(br d,J=9.8Hz,1H)1.12(d,J=7.0Hz,3H)1.08(s,9H).

[0277] Example 8

[0278]

[0279] Step 1: Synthesis of Compound 8-2

[0280] A 100 mL three-necked flask was used. 8-1 (2.2 g, 5.78 mmol) was dissolved in 30 mL of methanol, and then wet palladium on carbon (0.5 g, 10% palladium content) was added. The mixture was stirred at 25 °C for 16 h under a hydrogen atmosphere. After the reaction of the raw materials was complete, the resulting reaction solution was filtered through diatomaceous earth to obtain the crude product 8-2, which was then concentrated under reduced pressure.

[0281] MS–ESI m / z:[M+Na] + =405.1

[0282] Step 2: Synthesis of compound 8-3 hydrochloride

[0283] A 100 mL three-necked flask was used to dissolve 0.5 g (1.31 mmol) of 8-2 in 20 mL of ethyl acetate. Then, HCl / EtOAc (4 M, 3.26 mL) was added, and the mixture was stirred at 25 °C for 2 h. After the reaction of the starting materials was complete, the resulting reaction solution was concentrated under reduced pressure to obtain the crude product 8-3 hydrochloride.

[0284] MS–ESI m / z:[M+H] + =282.9.

[0285] Step 3: Synthesis of Compound 8-4

[0286] A 50 mL three-necked flask was used. 8-3 hydrochloride (471 mg, 1.67 mmol) was dissolved in dichloromethane (5 mL), followed by the addition of methyl trifluoroacetate (2.14 g, 16.68 mmol, 1.68 mL) and TEA (1.01 g, 10.01 mmol, 1.39 mL). The mixture was stirred at 38 °C for 12 hours. After the reaction was complete, the reaction solution was evaporated to dryness to obtain crude 8-4.

[0287] MS–ESI m / z:[M+H] + =379.1.

[0288] Step 4: Synthesis of Compounds 8-5

[0289] A 50 mL three-necked flask was used. 0.5 g (1.32 mmol) of 8-4 was dissolved in 10 mL of methanol. Then, lithium hydroxide monohydrate (110.90 mg, 2.64 mmol) and water (5 mL) were added, and the mixture was stirred at 25 °C for 12 hours. After the reaction was complete, 1 M hydrochloric acid was added to adjust the pH to 4–5. Then, 20 mL of water was added, and the mixture was extracted three times with 20 mL of ethyl acetate. The organic phase was retained, washed with saturated brine, dried over anhydrous sodium sulfate, and distilled under reduced pressure to obtain crude 8-5.

[0290] MS–ESI m / z:[M+H] + =365.0.

[0291] Step 5: Synthesis of Compound 8

[0292] Select a 50 mL three-necked flask, dissolve 8-5 (232 mg, 318.37 μmol, 50% purity) and 6-3 (208.09 mg, 318.37 μmol, 50% purity) in DMF (10 mL), then add HATU (242.11 mg, 636.73 μmol) and DIEA (123.44 mg, 955.10 μmol, 166.36 μL), and stir at 30 °C for 2 hours. After the reaction was complete, 30 mL of water was added to the resulting reaction solution and the mixture was extracted twice with 30 mL of ethyl acetate. The organic phase was retained, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated by vacuum distillation to obtain the crude product. The crude product was then separated by high performance liquid chromatography (column type: Phenomenex C18 80*40mm*3μm; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; ACN%: 45%-75%, 8 min) to obtain product 8.

[0293] MS–ESI m / z:[M+H] + =637.1.

[0294] 1 H NMR (400MHz, DMSO-d6) δppm 8.83 (br d, J=6.3Hz, 1H) 7.75 (dd, J=8.3, 4.6Hz, 1H) 7.67 (s, 1H) 7.17 (br s,3H)5.74-5.39(m,1H)4.58(s,1H)4.41(s,1H)4.22(s,1H)3.20(br t,J=9.1Hz,1H)3.15-3.01(m,1H)2.66(br s,1H)2.33(s,3H)2.13–1.97(m,2H)1.94-1.71(m,3H)1.37(br d,J=9.0Hz,1H)1.23(s,1H)0.97-1.02(s,9H)0.93-0.96(m,3H).

[0295] Example 9

[0296]

[0297] Synthesis route:

[0298]

[0299] Step 1: Synthesis of compound 9-4-1

[0300] The reaction flask was purged with nitrogen, and 4000 mL of n-hexane was added. Then, 1.05 L of diethylzinc solution (1 M n-hexane solution) was added. The mixture was cooled to 0 °C and stirred at 0 °C for 0.2 hr until the reaction system became clear. At 0 °C, 223.82 g of boron trifluoride diethyl ether (1.58 mol) was added and stirred at 0 °C for 0.5 hr. At 0 °C, 563.15 g of diiodomethane (2.10 mol) was added and stirred at 0 °C for 0.5 hr. Finally, 50 g of compound 8-1 (131.41 mmol) was added at 0 °C, and the mixture was heated to 40 °C and stirred for 16 hr. The reaction system was filtered, and the filter cake was washed with n-hexane (1500 mL). The filter cake was dissolved in ethyl acetate (5 L), and water (10 L) was added. 925 g of sodium carbonate was slowly added, followed by 1.85 kg of disodium ethylenediaminetetraacetate. The mixture was stirred for 15 min, allowed to stand for 30 min, and then separated. The aqueous phase was extracted once with ethyl acetate (2 L). The organic phases were combined and washed four times with saturated sodium chloride solution (3 L). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The unpurified solution was used directly in the next reaction to obtain compound 9-4-1.

[0301] MS–ESI m / z:[M+H] + =295.1.

[0302] Step 2: Synthesis of compound 9-4-2

[0303] Compound 9-4-1 (26 g, 78.59 mmol) was dissolved in tetrahydrofuran (260 mL) and water (85 mL). Sodium bicarbonate (13.20 g, 157.17 mmol) and di-tert-butyl dicarbonate (25.73 g, 117.88 mmol) were added, and the mixture was stirred at 20 °C for 16 h. The reaction mixture was extracted with ethyl acetate (200 mL), and the organic phase was washed with saturated sodium chloride solution (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was slurried in petroleum ether (5 V) for 30 min, filtered, and the filter cake was evaporated to dryness. Compound 9-4-2 was obtained.

[0304] MS–ESI m / z:[M+H] + =395.2.

[0305] Step 3: Synthesis of compound 9-4-3

[0306] Compound 9-4-2 (23 g, 58.30 mmol) was dissolved in tetrahydrofuran (140 mL), and water (50 mL), methanol (50 mL), and lithium hydroxide monohydrate (7.34 g, 174.90 mmol) were added. The mixture was stirred at 15 °C for 16 h. Ethyl acetate (200 mL) and water (50 mL) were added to the reaction solution, and the aqueous phase was separated. The aqueous phase was extracted with 5% citric acid (100 mL) and ethyl acetate (200 mL × 2). The organic phase was washed with saturated sodium chloride solution (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 9-4-3.

[0307] MS–ESI m / z:[M+H] + =381.1.

[0308] Step 4: Synthesis of the hydrochloride salt of compound 9-4-4

[0309] Compound 9-4-3 (10 g, 26.28 mmol) was dissolved in dichloromethane (38 mL), and hydrogen chloride / dioxane (4 M, 32.85 mL) was added. The reaction was carried out at 20 °C for 16 h. The solution was concentrated under reduced pressure to give the hydrochloride salt of compound 9-4-4.

[0310] MS–ESI m / z:[M+H] + =281.2.

[0311] Step 5: Synthesis of compound 9-4

[0312] The hydrochloride salt of compound 9-4-4 (8.3 g, 26.20 mmol) was dissolved in methanol (83 mL), and triethylamine (7.95 g, 78.59 mmol, 10.94 mL) and methyl trifluoroacetate (13.42 g, 104.79 mmol) were added. The mixture was stirred at 40 °C for 16 h. The solution was evaporated to dryness, dissolved in ethyl acetate (10 mL) and water (10 mL), and separated. The aqueous phase was extracted with ethyl acetate (10 mL × 2). The combined organic phases were washed with 5% citric acid (20 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness. A slurry of ethyl acetate and petroleum ether at a ratio of 1:5 (5V) was prepared, filtered, and evaporated to dryness. Compound 9-4 was obtained.

[0313] MS–ESI m / z:[M+H] + =377.3.

[0314] Step 6: Synthesis of Compound 9

[0315] Compound 9-4 (95.59 mg, 253.98 μmol) was dissolved in DMF (5 mL), and DIEA (65.65 mg, 507.95 μmol, 88.48 μL) was added. The mixture was stirred at 25 °C for 5 minutes. Then, a DMF (3 mL) solution of HATU (289.71 mg, 761.93 μmol) and 6-3 hydrochloride (83 mg, 253.98 μmol) was added to the reaction solution. After the reaction was complete, water was added to the reaction solution, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine (30 mL x 3), dried over anhydrous magnesium sulfate, filtered, and the filtrate was concentrated under reduced pressure to remove the solvent. The residue was separated by high performance liquid chromatography [column type: Phenomenex C1880*40mm*3μm; mobile phase: [water (containing 0.1% (ammonia water + ammonium bicarbonate)) - acetonitrile]; acetonitrile %: 43%-73%, 8min] to obtain compound 9.

[0316] MS–ESI m / z:[M+H] + =649.1.

[0317] 1 H NMR (400MHz, DMSO-d6) δppm 8.83 (br d, J = 3.51Hz, 1H) 8.78 (br d, J = 7.78Hz, 1H) 8.65 (d, J = 8.28Hz, 1H) 7.79-7.70 (m, 1H) 7.66 (s, 1H) 5.59 (br t,J=8.41Hz,1H)4.64(br s,1H)4.22(s,1H)3.25-3.15(m,1H)3.14-3.07(m,1H)2.67(br s,1H)2.44-2.21(m,2H)2.13-1.94(m,2H)1.89-1.62(m,4H)1.59-1.44(m,1H)1.23(br s,2H)1.04–0.83(m,9H)0.78(br s,1H)0.67(br d,J=3.26Hz,1H)0.52-0.31(m,2H).

[0318] Biological testing:

[0319] Experimental Example 1: Evaluation of the in vitro anti-novel coronavirus Mpro protease activity of the test compound

[0320] 1. Experimental materials:

[0321] 1.1 Reagents and Consumables:

[0322] Table 1. Reagents, consumables and their brands

[0323]

[0324] 1.2 Instruments:

[0325] Table 2. Instruments and their brands

[0326] instrument brand 1 SpectraMax M2e microplate reader Molecular Devices 2 Echo 655 Liquid Workstation Labcyte 3 benchtop high-speed centrifuge Eppendorf

[0327] 2. Experimental Methods:

[0328] The compound was dissolved in DMSO and serially diluted 3-fold using Echo 655 according to the required concentration, resulting in 10 concentration points, with double replicates for each concentration, added to 384-well plates. Mpro protein and substrate were diluted with test buffer (100 mM NaCl, 20 mM Tris-HCl, 1 mM EDTA). Mpro protein was added to the 384-well plate and incubated with the compound at room temperature for 30 min. Then, the substrate was added, with a test concentration of 25 nM for Mpro protein and 25 μM for the substrate. The plate was incubated at 30°C for 60 min. The fluorescence signal at Ex / Em = 340 nm / 490 nm was then detected using a microplate reader. Background wells containing both substrate and compound but without Mpro protein served as controls.

[0329] 3. Data Analysis:

[0330] 1) Calculate the inhibition rate using the following formula:

[0331] Inhibition rate % = [(compound - BG)] 化合物 )-(ZPE-BG ZPE )] / [(HPE-BG HPE )-(ZPE-BG ZPE )]*100%

[0332] # HPE: 100% inhibition control, containing 25 nM Mpro protein + 25 μM substrate + 1 μM GC376

[0333] ZPE: Non-inhibitory control, containing 25 nM Mpro protein + 25 μM substrate, free of compounds.

[0334] Compounds: Test compound wells containing 25 nM Mpro protein + 25 μM substrate + compound

[0335] BG: Background control wells containing 25 μM substrate + compound, free of Mpro protein.

[0336] 2) The inhibition rate data (inhibition rate %) of the compounds were analyzed using GraphPad Prism software using a log(agonist) vs. res-nse-variable slope nonlinear fitting analysis to obtain the IC50 of the compounds. 50 Values. The experimental results are shown in Table 3:

[0337] Table 3: In vitro anti-novel coronavirus Mpro protease activity of the test compounds

[0338] Compound numbering <![CDATA[IC 50 (nM) <!-- 29 -->]]> 1 74 2 38 3 15.1 4 10 5 14 7 11 8 47 9 10

[0339] Conclusion: The compounds of this invention exhibit good in vitro activity against the novel coronavirus Mpro protease.

[0340] Experimental Example 2: Evaluation of the in vitro anticoronavirus activity of compounds using a cytopathic model

[0341] 1. Experimental Materials

[0342] 1.1. Reagents and Consumables

[0343] Table 4. Reagents, Consumables and Their Names

[0344] Names of reagents and consumables brand 1 MEM culture medium Sigma 2 L-Glutamine Gibco 3 Non-essential amino acids Gibco 4 Penicillin-Streptomycin Solution HyClone 5 Fetal bovine serum (FBS) ExCell 6 Phosphate-buffered saline (DPBS) Corning 7 0.25% pancreatic enzyme Gibco 8 CellTiter Glo Cell Viability Assay Kit Promega 9 Remdesivir MCE 10 96-hole plate Grenier

[0345] 1.2. Instruments

[0346] Table 5. Instruments and their brands

[0347] instrument brand 1 ELISA reader BioTek 2 Cell counter Beckman 3 <![CDATA[CO2 Incubator]]> Thermo

[0348] 1.3. Cells and Viruses

[0349] MRC5 cells and coronavirus HCoV OC43 were purchased from ATCC.

[0350] MRC5 cells were cultured in MEM (Sigma) medium supplemented with 10% fetal bovine serum (Excell), 1% penicillin-dextrin (Hyclone), 1% L-glutamine (Gibco), and 1% non-essential amino acids (Gibco). MEM (Sigma) medium supplemented with 5% fetal bovine serum (Excell), 1% penicillin-dextrin (Hyclone), 1% L-glutamine (Gibco), and 1% non-essential amino acids (Gibco) was used as the experimental culture medium.

[0351] 2. Experimental Methods

[0352] Table 6 Virus testing methods used in this study

[0353]

[0354] Cells were seeded at a specific density (Table 2) into 96-well microplates and incubated overnight in a 5% CO2, 37°C incubator. The next day, serially diluted compounds (8 concentration points, duplicate wells) were added at 50 μL per well. Subsequently, diluted virus was added at 100 TCID⁻¹ per well. 50 Cells were added at a density of 50 μL per well. Cell controls (cells, no compound treatment or viral infection), virus controls (cells infected with virus, no compound treatment), and culture medium controls (culture medium only) were set up. The final culture medium volume for this experiment was 200 μL, and the final concentration of DMSO in the culture medium was 0.5%. Cells were cultured for 5 days in a 5% CO2 incubator at 33°C. Cell viability was assessed using the CellTiter Glo (Promega) cell viability assay kit. The cytotoxicity assay was performed under the same conditions as the antiviral assay, but without viral infection.

[0355] 3. Data Analysis

[0356] The antiviral activity and cytotoxicity of the compounds were expressed as the inhibition rate (%) and cell viability (%) of the compounds at different concentrations against virus-induced cytopathic effects, respectively. The calculation formulas are as follows:

[0357] Inhibition rate (%) = (Test well reading - Average value of virus control) / (Average value of cell control - Average value of virus control) × 100

[0358] Cell viability (%) = (Test well reading - Average value of culture medium control) / (Average value of cell control - Average value of culture medium control) × 100

[0359] The inhibition rate and cell viability of the compound were analyzed using nonlinear fitting analysis with GraphPad Prism to calculate the half-maximal effective concentration (EC50) of the compound. 50 ) and half-maximal cytotoxic concentration (CC) 50 )value.

[0360] Conclusion: The compounds of this invention exhibit good in vitro anti-coronavirus activity at the cellular level and are non-cytotoxic.

[0361] Experiment Example 3: Evaluation of the in vivo antiviral efficacy of the test compound using a neonatal rat infection model of coronavirus OC43 virus strain.

[0362] C57BL / 6J suckling mice were infected with a lethal dose of coronavirus via nasal instillation. Two hours prior to infection, the mice were treated with a solvent (5% DMSO + 40% PEG400 + 55% water) and the compound of this invention. During the experiment, the mice's weight, health status, and survival were monitored daily to evaluate the protective effect of the compound of this invention at different doses.

[0363] In the solvent group, the weight of suckling mice continued to decrease 6 days after viral inoculation, with a final survival rate of 0%. Compound 9 (25 mpk) was first administered 2 hours before infection, and the final survival rate was 83.33%. The experimental results are shown in Table 7.

[0364] Table 7. Mouse survival analysis

[0365]

[0366] Experimental conclusion: The compound of this invention has good in vivo anti-coronavirus efficacy.

Claims

1. A compound of Formula (II) or a pharmaceutically acceptable salt thereof, ； wherein, R1is each independently selected from the group consisting of halogen, CN, OH, NH2, C 1-3 alkyl and C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy are each independently optionally substituted with 1, 2, or 3 halogen; or, two R1and the atoms to which they are attached form a C 3-6 cycloalkyl, said C 3-6 cycloalkyl is optionally substituted with 1, 2, or 3 R a substituents; R a each independently is selected from the group consisting of halogen and C 1-3 alkyl; n is selected from 0, 1, 2, and 3; R2is selected from and ; R3is selected from C 1-3 alkyl, C 1-3 alkoxy, -CH2R4and -CH2OR4, said C 1-3 alkyl and C 1-3 each alkyl and C R4 is selected from phenyl and 5-6 membered heteroaryl, optionally independently substituted with 1, 2, or 3 R; R is selected from the group consisting of halogen and C 1-3 alkyl; T1 is selected from O and S; A1, A2, A3, and A4 are each independently selected from N and CH; Ring A is selected from and .

2. The compound or pharmaceutically acceptable salt thereof of claim 1, wherein, R1 is selected from F and methyl.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, two R1and the atoms to which they are attached form C 3-6 cycloalkyl.

4. The compound or pharmaceutically acceptable salt thereof of claim 3, wherein, two R1 and the atoms to which they are attached form a cyclopropyl.

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, Structural unit selected from , , , , and .

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, Structural unit selected from , , , , and .

7. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, R3 is selected from -CF3.

8. The compound of claim 1 or a pharmaceutically acceptable salt thereof, selected from the structures of Formulae (II-2) and (II-3), ； wherein, selected from and ; R1, A1, A2, A3, A4, T1, and n are as defined in claim 1.

9. A compound of the following formula or a pharmaceutically acceptable salt thereof, , , , , , , , , , , , , , , , , and .

10. A pharmaceutical composition comprising a therapeutically effective amount of a compound of any one of claims 1-9 or a pharmaceutically acceptable salt thereof.

11. Use of a compound of any one of claims 1-9 or a pharmaceutically acceptable salt thereof or a composition of claim 10 for the manufacture of a medicament for the treatment of a coronavirus infection.

12. The use of claim 11, wherein the coronavirus is selected from HCoV OC43.

13. A compound of Formula (I) or a pharmaceutically acceptable salt thereof, ； wherein, R1is each independently selected from the group consisting of halogen, CN, OH, NH2, C 1-3 alkyl or C 1-3 alkoxy, said C 1-3 alkyl and C 1-3 alkoxy are each independently optionally substituted with 1, 2, or 3 halogen; or, two R1and the atoms to which they are attached form a C 3-6 cycloalkyl, said C 3-6 cycloalkyl is optionally substituted with 1, 2, or 3 R a substituents; n is selected from 0, 1, 2, or 3; R a each independently is selected from halogen or C 1-3 alkyl; R2is selected from tert-butyl, C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, or phenyl, said C 3-10 cycloalkyl, 3-10 membered heterocycloalkyl, and phenyl are optionally substituted with 1, 2, or 3 R b substituents; R b each independently is selected from halogen or C 1-3 alkyl; R3is selected from C 1-3 alkyl, C 1-3 alkoxy, -CH2R4or -CH2OR4, said C 1-3 alkyl and C 1-3 alkoxy is optionally substituted with 1, 2 or 3 halo; R4 is selected from phenyl or 5-6 membered heteroaryl, optionally independently substituted with 1, 2, or 3 R; R is selected from halo or C 1-3 alkyl; A1, A2, A3, and A4 are each independently selected from N or CH, and 0, 1, or 2 of A1, A2, A3, and A4 are selected from N, and the remainder are selected from CH.

14. The compound or pharmaceutically acceptable salt thereof of claim 13, wherein, R1 is selected from F or methyl.

15. The compound of claim 13, or a pharmaceutically acceptable salt thereof, wherein, two R1and the atoms to which they are attached form C 3-6 cycloalkyl.

16. The compound or pharmaceutically acceptable salt thereof of claim 15, wherein, two R1 and the atoms to which they are attached form a cyclopropyl.

17. The compound of claim 13, or a pharmaceutically acceptable salt thereof, wherein, structural unit selected from , , or .

18. The compound or pharmaceutically acceptable salt thereof of claim 13, wherein, R2 is selected from tert-butyl.

19. The compound or pharmaceutically acceptable salt thereof of claim 13, wherein, R3 is selected from -CF3.

20. The compound of any one of claims 13-17 or a pharmaceutically acceptable salt thereof, selected from the structure of Formula (I-1), ； wherein R1 and n are as defined in any one of claims 13-17.

21. A compound of the following formula or a pharmaceutically acceptable salt thereof, 。

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