Ketone amide derivatives and uses thereof

Abstract

Disclosed are ketone amide derivatives and applications thereof, and specifically disclosed are compounds of formula (IV) and pharmaceutically acceptable salts thereof.

Description

Technical Field

[0001] This invention relates to a class of ketoamide derivatives and their applications, specifically to compounds of formula (IV) and their pharmaceutically acceptable salts. Background Technology

[0002] The main protease of coronaviruses, also known as the 3CL protease, is a key protein in viral replication, primarily functioning to hydrolyze two polyproteins expressed by the virus. Sequence analysis suggests that the 3CL protease may be a key target for drug design. Developing its inhibitors to block coronavirus replication is of significant value and importance for the prevention and control of coronavirus infection.

[0003] PF-07321332 is a potent orally active SARS-CoV3CL PRO inhibitor, with the structure shown below:

[0004] . Summary of the Invention

[0005] This invention provides compounds of formula (IV) or pharmaceutically acceptable salts thereof, selected from:

[0006]

[0007] in,

[0008] R3 is selected from and R4;

[0009] R1 is independently selected from F, Cl, Br, I, and OR. 11 CN, CH3S(O) m -and NHR 12 and C 1-3 Alkyl, the C 1-3 The alkyl group may be optionally substituted with 1, 2 or 3 F atoms;

[0010] R 11 Selected from H, C 1-3 Alkyl group, CH3(OCH2CH2) p - and H(OCH2CH2) q -, the C 1-3 The alkyl group may be optionally substituted with 1, 2 or 3 F atoms;

[0011] R 12 Selected from C 1-3 Alkyl, CH3CO- and CH3SO2-, wherein the C 1-3 Alkyl groups, CH3CO-, and CH3SO2- may be independently substituted with 1, 2, or 3 F atoms;

[0012] m is selected from 0, 1, and 2;

[0013] p and q are selected from 1, 2, 3, 4, 5, and 6;

[0014] n is selected from 0, 1, 2, 3, and 4;

[0015] R2 is selected from C 1-4 Alkyl, C 3-6 cycloalkyl and benzyl, the C 1-4 Alkyl, C 3-6 The cycloalkyl and benzyl groups may be optionally substituted with 1, 2 or 3 F groups;

[0016] R4 is selected from C, which can be replaced by 1, 2, or 3 Fs. 1-8 alkyl;

[0017] Ring A is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl and phenyl;

[0018] Ring B is selected from C 3-8 Cycloalkyl and 5-membered heterocycloalkyl; the C 3-8 Cycloalkyl and 5-membered heterocycloalkyl groups are optionally surrounded by 1 or 2 R groups. a replace;

[0019] R a Each is independently selected from H and C. 1-3 alkyl;

[0020] The “5-membered heterocyclic alkyl” comprises 1, 2 or 3 heteroatoms or groups independently selected from O, S, SO2, N, P and Se.

[0021] In some embodiments of the present invention, R1 is selected from F and methyl, and other variables are as defined in the present invention.

[0022] In some embodiments of the present invention, the ring A is selected from... , , , and Other variables are as defined in this invention.

[0023] In some embodiments of the present invention, the above-mentioned structural unit Selected from , , , , and Other variables are as defined in this invention.

[0024] In some embodiments of the present invention, R4 is selected from tert-butyl, and other variables are as defined in the present invention.

[0025] In some embodiments of the present invention, the above-mentioned Ra Selected from H and methyl, other variables are as defined in this invention.

[0026] In some embodiments of the present invention, the ring B is selected from... and Other variables are as defined in this invention.

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

[0028] This invention provides compounds of the following formula or pharmaceutically acceptable salts thereof, selected from:

[0029]

[0030] in,

[0031] R1 is independently selected from F, Cl, Br, I, and OR. 11 CN, CH3S(O) m -and NHR 12 and C 1-3 Alkyl, the C 1-3 The alkyl group may be optionally substituted with 1, 2 or 3 F atoms;

[0032] R 11 Selected from H, C 1-3 Alkyl group, CH3(OCH2CH2) p - and H(OCH2CH2) q -, the C 1-3 The alkyl group may be optionally substituted with 1, 2 or 3 F atoms;

[0033] R 12 Selected from C 1-3 Alkyl, CH3CO- and CH3SO2-, wherein the C 1-3 The alkyl group may be optionally substituted with 1, 2 or 3 F atoms;

[0034] m is selected from 0, 1, and 2;

[0035] p and q are selected from 1, 2, 3, 4, 5, and 6;

[0036] n is selected from 0, 1, 2, 3, and 4;

[0037] R2 is selected from C 1-4 Alkyl, C 3-6 cycloalkyl and benzyl, the C 1-4 Alkyl, C 3-6 The cycloalkyl and benzyl groups may be optionally substituted with 1, 2 or 3 F groups;

[0038] Ring A is selected from C 3-10 Cycloalkyl, 3-10 membered heterocyclic alkyl and phenyl;

[0039] Ring B is selected from C 3-6 Cycloalkyl and 5-membered heterocycloalkyl; the C 3-6 Cycloalkyl and 5-membered heterocycloalkyl groups are optionally surrounded by 1 or 2 R groups. a replace;

[0040] R a Each is independently selected from H and C. 1-3 alkyl;

[0041] The “5-membered heterocyclic alkyl” comprises 1, 2 or 3 heteroatoms or groups independently selected from O, S, SO2, N, P and Se.

[0042] In some embodiments of the present invention, R1 is selected from F and methyl, and other variables are as defined in the present invention.

[0043] In some embodiments of the present invention, the ring A is selected from... , , , and Other variables are as defined in this invention.

[0044] In some embodiments of the present invention, the above-mentioned structural unit Selected from , , , and Other variables are as defined in this invention.

[0045] In some embodiments of the present invention, the above-mentioned R a Selected from H and methyl, other variables are as defined in this invention.

[0046] In some embodiments of the present invention, the above-mentioned structural unit Selected from or Other variables are as defined in this invention.

[0047] In some embodiments of the present invention, the compounds are selected from the structures shown in formulas (I-1), (IV-1), and (IV-2).

[0048] , ,

[0049] ,

[0050] Wherein, R1, R2, R3, n and ring A are as defined in this invention.

[0051] In some embodiments of the present invention, the compounds are selected from the structures shown in formulas (I-1a), (IV-1a), and (IV-2a).

[0052] , , ,

[0053] Wherein, R1, R2, R3, n and ring A are as defined in this invention.

[0054] Some solutions in this invention are derived from arbitrary combinations of the above-mentioned variables.

[0055] This invention also provides the following compounds or pharmaceutically acceptable salts thereof, selected from:

[0056] .

[0057] This invention also provides the following compounds or pharmaceutically acceptable salts thereof, selected from:

[0058] .

[0059] The present invention provides a method of combined administration, comprising administering to a subject requiring treatment a therapeutically effective amount of any of the compounds of the present invention or a pharmaceutically acceptable salt thereof, and a therapeutically acceptable dose of another antiviral drug.

[0060] The present invention also provides a method for treating coronavirus infection, the method comprising administering to a subject requiring the treatment a therapeutically effective amount of any of the compounds of the present invention or a pharmaceutically acceptable salt thereof, and a dose of other therapeutically acceptable antiviral drugs.

[0061] In some embodiments of the present invention, the other antiviral drugs mentioned above are ritonavir, indinavir, nelfinavir, saquinavir, amprenavir, or lopinavir. In the aforementioned method for treating coronavirus infection, the mass ratio of the compound or its pharmaceutically acceptable salt described in any of the technical solutions of the present invention to ritonavir, indinavir, nelfinavir, saquinavir, amprenavir, or lopinavir is 1:1 to 5:1, specifically 1:1, 2:1, 3:1, 4:1, or 5:1, etc. Experiments have surprisingly shown that the two therapeutic components within this ratio range are beneficial for achieving synergistic effects, resulting in better overall therapeutic efficacy. Furthermore, the method can administer the therapeutic components contained in the same unit formulation, i.e., compound formulation administration, or it can administer formulations containing different therapeutic components separately, i.e., clinical combination therapy.

[0062] In some embodiments of the present invention, the aforementioned coronavirus infection is HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, or SARS-CoV-2 and its variants.

[0063] In some embodiments of the present invention, the aforementioned coronavirus infection is SARS-CoV-2 and its variants.

[0064] The present invention also provides the following synthetic route:

[0065]

[0066] Technical effect

[0067] 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 no cytotoxicity. In pharmacokinetic studies, the compounds of this invention showed significantly higher plasma exposure, slower clearance rate, and longer half-life compared to the reference molecule PF-07321332, demonstrating better pharmacokinetic properties.

[0068] Definitions and Explanations

[0069] 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.

[0070] 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.

[0071] 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, base addition salts can be obtained by contacting a neutral form of such compound with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts or similar salts. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting a neutral form of such compound with a sufficient amount of acid in a pure solution or a suitable inert solvent.

[0072] 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.

[0073] "Pharmaceutical composition" means containing one or more of the compounds described in this application, their isomers or pharmaceutically acceptable salts thereof, and other components such as physiologically / pharmaceuticalally 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.

[0074] The term "therapeutic effective amount" refers to an amount of a compound, when administered, sufficient to stop or slow the progression of one or more symptoms or conditions of a disease to a certain extent. The term "therapeutic effective amount" also refers to an amount of a compound sufficient to detect a biological or pharmaceutical response (e.g., protein, enzyme, RNA, or DNA) in a biomolecule, cell, tissue, system, animal, or human. This response is desired by researchers, veterinarians, physicians, or clinicians.

[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 envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, ( R )- and(S - Enantiomers, diastereomers, ( D )-Isomer, ( L (Isomers, racemic mixtures thereof, and other mixtures, such as mixtures enriched with enantiomers or diastereomers, are all 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 ( ) represents the absolute configuration of a solid center, using a straight solid line key ( ) and straight dashed key ( The relative configuration of the center of a solid is represented by a wavy line ( ). ) indicates a 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 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%.

[0083] 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%.

[0084] Optically active materials can be prepared through chiral synthesis, chiral reagents, or other conventional techniques. R )-and( S )-Isomers and D and L Isomers. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide a pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, and then the diastereomeric isomer is resolved by conventional methods known in the art, and the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0085] 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.

[0086] 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.

[0087] 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 resulting 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.

[0088] 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.

[0089] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0090] 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.

[0091] 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.

[0092] 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.

[0093] 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. or 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.

[0094] 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 the linking group, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0095] 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 lines ( () indicates that the oxygen atom in the group is bonded to another group. For example, a straight solid line bond in -OCH3 indicates that the oxygen atom in the group is bonded to another group. 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.

[0096] 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.

[0097] 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 9-12Similarly, 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.

[0098] Unless otherwise specified, the term "C" 1-8 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 8 carbon atoms. The C 1-8 Alkyl groups include C 1-6 C 1-5 C 1-4 C 1-3 C 1-2 C 2-6 C 2-4 C8, C7, C6, and C5 alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). 1-8 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), and propyl (including...). n -propyl and isopropyl), butyl (including n -Butyl, Isobutyl s -Butyl and t -Butyl), pentyl (including n -pentyl, isopentyl and neopentyl), hexyl, heptyl, octyl, etc.

[0099] Unless otherwise specified, the term "C" 1-4 "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 4 carbon atoms. The C 1-4 Alkyl groups include C 1-2 C 1-3 and C 2-3 Alkyl groups, etc.; they can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-4 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), and propyl (including...). n -propyl and isopropyl), butyl (including n -Butyl, Isobutyl s -Butyl and t (-Butyl) etc.

[0100] 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 C1-2 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), and propyl (including...). n -propyl and isopropyl, etc.

[0101] 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, norbornelalkyl, [2.2.2]bicyclooctane, [4.4.0]bicyclodecane, etc.

[0102] Unless otherwise specified, "C 3-8 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 8 carbon atoms, including monocyclic, bicyclic, and tricyclic systems, wherein bicyclic and tricyclic systems include spirocyclic, fused, and bridged rings. The C 3-8 Cycloalkyl groups include C 3-8 C 3-6 C 3-5 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 3-8 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, spiro[2.4]cyclohexane, 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 3-4 C 3-5 C 4-5 C 5-8Or 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, the term "3-10 membered heterocyclic alkyl" on its own or in combination with other terms 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, and 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 3-9, 3-8, 3-6, 5-9, 5, 6, 7, 8, and 9 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.

[0105] Unless otherwise specified, the term "5-membered heterocyclic alkyl" on its own or in combination with other terms refers to a saturated cyclic group consisting of 5 ring atoms, wherein 1, 2, or 3 of the ring atoms are heteroatoms independently selected from O, S, N, P, and 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). Examples of 5-membered heterocyclic alkyl groups include, but are not limited to, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, etc.

[0106] 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.

[0107] The structures of the compounds of this invention can be confirmed using conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed using conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) can be used. Diffraction intensity data of the grown single crystals are collected using a Bruker D8 venture diffractometer with CuKα radiation as the light source and a φ / ω scan mode. After collecting the relevant data, the crystal structure can be further analyzed using the direct method (Shelxs 97) to confirm the absolute configuration.

[0108] The solvent used in this invention is commercially available.

[0109] This invention uses the following abbreviations:

[0110] ACN represents acetonitrile; Boc represents tert-butyloxycarbonyl; Bn represents benzyl; DCM represents dichloromethane; DMSO represents dimethyl sulfoxide; ℃ represents degrees Celsius; hr represents hours; LiBH4 represents sodium borohydride; THF represents tetrahydrofuran; Ts represents p-toluenesulfonyl; Ac represents acetyl; Me represents methyl; Et represents ethyl.

[0111] Compounds are named according to conventional naming principles in the art or using ChemDraw® software; commercially available compounds are named according to the supplier catalog. Detailed Implementation

[0112] 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.

[0113] Example 1

[0114]

[0115] Synthesis route:

[0116]

[0117] Step 1: Synthesis of hydrochloride salts of compounds 1-2

[0118] Compound 1-1 (500 mg, 1.75 mmol) was dissolved in ethyl acetate (5 mL), and a solution of hydrogen chloride in ethyl acetate (10 mL, 4 N) was added. The reaction was stirred at 20 °C for 2 hours. The solution was concentrated under reduced pressure without purification to give the hydrochloride salt of compound 1-2. 1 H NMR (400 MHz, CD3OD) δ = 4.28 - 4.20 (m, 1H), 3.91 - 3.81 (m, 3H), 3.45- 3.35 (m, 2H), 2.86 - 2.74 (m, 1H), 2.48 - 2.36 (m, 1H), 2.29 - 2.19 (m,1H), 2.02 - 1.94 (m, 1H), 1.93 - 1.80 (m, 1H).

[0119] Step 2: Synthesis of compounds 1-4

[0120] The compound Boc-L-cyclohexylglycine (1 g, 3.89 mmol) was added to N,N-dimethylformamide (10 mL), followed by the addition of 2-(7-azobenzotriazole)- N,N,N,N Tetramethylurea hexafluorophosphate (1.77 g, 4.66 mmol) was reacted and stirred for 0.5 h. Diisopropylethylamine (1.26 g, 9.72 mmol) and the hydrochloride salts of compounds 1-3 (1.02 g, 4.66 mmol) were added, and the reaction was stirred at 20 °C for 16 h. Methyl tert-butyl ether (50 mL) was added to the reaction mixture, followed by washing with water (20 mL), 3% citric acid (20 mL × 2), and saturated sodium chloride solution (20 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) yielded compounds 1-4. 1 H NMR (400MHz, CDCl3) δ = 5.22 - 5.11 (m, 1H), 4.36 (d, J =3.9 Hz, 1H), 4.27 (dd, J =6.9, 9.3Hz, 1H), 4.21 - 4.12 (m, 2H), 3.83 (dd, J =7.8, 10.4 Hz, 1H), 3.70 (br dd, J=3.6, 10.4 Hz, 1H), 2.81 - 2.61 (m, 2H), 1.82 - 1.70 (m, 6H), 1.68 - 1.61 (m,4H), 1.56 - 1.48 (m, 2H), 1.46 - 1.38 (m, 9H), 1.29 - 1.22 (m, 4H), 1.21 -0.98 (m, 4H).

[0121] Step 3: Synthesis of compounds 1-5

[0122] Compounds 1-4 (1.41 g, 3.34 mmol) were added to tetrahydrofuran (14 mL), followed by a 5 mL solution of lithium hydroxide monohydrate (LiOH·H₂O) (280.03 mg, 6.67 mmol) in water. The reaction was stirred at 20 °C for 16 hours. The crude product was neutralized with 3% citric acid solution (50 mL), extracted with ethyl acetate (50 mL), and the organic phase was washed with saturated sodium chloride solution (30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Without purification, compounds 1-5 were obtained. 1 H NMR (400MHz, DMSO-d6) δ = 12.58 - 12.23 (m, 1H), 6.92 - 6.82 (m, 1H), 4.11 - 3.94 (m, 2H), 3.82 - 3.76 (m, 1H), 3.72 - 3.62 (m, 1H), 2.73 - 2.64 (m, 1H), 2.62 - 2.55 (m, 1H), 1.92 - 1.42 (m, 12H), 1.40 - 1.32 (m, 9H), 1.18 - 1.06 (m, 3H), 1.00- 0.81 (m, 2H).

[0123] Step 4: Synthesis of compounds 1-6

[0124] Compounds 1-5 (650 mg, 1.65 mmol) were added to 2-butanone (7 mL), followed by 1-hydroxybenzotriazole (222.63 mg, 1.65 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (379.03 mg, 1.98 mmol), and diisopropylethylamine (638.84 mg, 4.94 mmol). The reaction mixture was stirred at 20 °C for 0.5 hr. Then, the hydrochloride of compound 1-2 (366.88 mg, 1.65 mmol) was added, and the reaction mixture was stirred at 20 °C for 16 h. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane:methanol (30 mL × 2, 10:1). The organic phases were combined, washed with 3% citric acid (20 mL × 2), washed with saturated sodium chloride solution (20 mL), and dried over anhydrous sodium sulfate. The mixture was then filtered and concentrated under reduced pressure. Compounds 1-6 were obtained by silica gel column chromatography (dichloromethane:methanol = 20:1). 1 H NMR (400 MHz, CDCl3) δ = 7.49 - 7.42(m, 1H), 6.23 - 6.05 (m, 1H), 5.28 - 5.17 (m, 1H), 4.64 - 4.51 (m, 1H), 4.43- 4.24 (m, 2H), 3.92 - 3.81 (m, 1H), 3.78 - 3.70 (m, 3H), 3.39 - 3.27 (m,2H), 2.94 - 2.75 (m, 2H), 2.57 - 2.36 (m, 2H), 2.24 - 2.07 (m, 1H), 1.94 -1.50 (m, 14H), 1.49 - 1.41 (m, 9H), 1.27 - 0.95 (m, 6H).

[0125] Step 5: Synthesis of compounds 1-7

[0126] Compounds 1-6 (3.10 g, 5.51 mmol) were dissolved in tetrahydrofuran (31 mL), and lithium borohydride (240.02 mg, 11.02 mmol) was added at 0 °C. The mixture was then slowly heated to 20 °C and reacted for 2 h. Water (10 mL) and ethyl acetate (20 mL) were added to the reaction solution, and the mixture was stirred for 10 min. A white solid precipitated out, and the mixture was filtered to obtain the filter cake, which was the crude product 1-7. [M+1]+ = 535.4.

[0127] Step 6: Synthesis of compounds 1-8

[0128] Compounds 1-7 (0.5 g, 935.13 μmol) were dissolved in dichloromethane (10 mL), and then Dys-Martin oxidant (594.94 mg, 1.40 mmol) was added to the reaction system. The reaction was stirred at 25 °C for 16 h. Saturated sodium thiosulfate (15 mL) and saturated sodium bicarbonate solution (15 mL) were added to the reaction system, and the mixture was stirred for 10 min. The mixture was extracted with dichloromethane (50 mL × 2), and the organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude compounds 1-8. [M+1]+ = 533.4.

[0129] Step 7: Synthesis of compounds 1-9

[0130] Compounds 1-8 (436 mg, 818.52 μmol) were dissolved in dichloromethane (5 mL), and glacial acetic acid (58.98 mg, 982.22 mmol) and cyclopentyl isocyanate (94.44 mg, 982.22 μmol) were added to the reaction system. The reaction was stirred at 25 °C for 2 h. A saturated ammonium chloride solution (10 mL) was added to the reaction system and stirred for 10 min. Dichloromethane (20 mL) was added for extraction. The organic phase was washed with water (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification was achieved by silica gel column chromatography (dichloromethane:methanol = 10:1) to give compounds 1-9. [M+1]+ = 688.4.

[0131] Step 8: Synthesis of compounds 1-10

[0132] Compounds 1-9 (190 mg, 276.22 μmol) were dissolved in methanol (3 mL), followed by a solution of potassium carbonate (95.44 mg, 690.54 μmol) in water (2 mL). The reaction was stirred at 20 °C for 16 h. 3% citric acid (20 mL) was added to the reaction mixture, and the mixture was extracted three times with dichloromethane (40 mL). The organic phase was washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude 1-10. [M+1] + = 646.5.

[0133] Step 9: Synthesis of compounds 1-11

[0134] Compound 1-10 (238.00 mg, 368.52 μmol) was dissolved in dichloromethane (24 mL), and then Dysmartin oxidant (203.19 mg, 479.08 μmol) was added. The reaction was stirred at 20 °C for 18 h. Sodium thiosulfate (15 mL) and sodium bicarbonate solution (15 mL) were added to the reaction system, and the mixture was stirred for 10 min. Extraction was performed with dichloromethane (50 mL × 2). The organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was achieved by silica gel column chromatography (dichloromethane:methanol = 20:1) to give product 1-11. [M+1] + = 644.5.

[0135] Step 10: Synthesis of compounds 1-12

[0136] Compound 1-11 (125 mg, 194.16 μmol) was dissolved in tetrahydrofuran (3 mL), and then ethyl acetate hydrochloride (4 M, 2.91 mL) was added. The reaction was stirred at 20 °C for 1 h. The reaction solution was directly rotary evaporated using an oil pump, and the rotary evaporation was repeated with a small amount of dichloromethane to give compound 1-12. [M+1] + =544.4.

[0137] Step 11: Synthesis of Compound 1

[0138] Compounds 1-12 (125 mg, 229.91 μmol) were dissolved in tetrahydrofuran (2.5 mL). Trifluoroacetic anhydride (193.15 mg, 919.63 μmol) and pyridine (127.30 mg, 1.61 mmol) were added at 0 °C. The reaction mixture was stirred at 20 °C for 16 h. Water (20 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (40 mL × 2). The organic phase was washed successively with 3% citric acid (40 mL) and saturated sodium chloride solution (40 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by preparative HPLC to obtain compound 1. [M+1] + = 640.0; 1H NMR (400 MHz, CD3OD) δ ppm 0.94 - 1.10 (m, 2 H),1.13 - 1.32 (m, 3 H) 1.32 - 1.46 (m, 1 H), 1.47 - 1.57 (m, 3 H), 1.59 - 1.68(m, 4 H), 1.69 - 1.81 (m, 6 H), 1.83 - 2.00 (m, 5 H), 2.01 - 2.17 (m, 1 H), 2.19 - 2.38 (m, 1 H), 2.49 - 2.57 (m, 1 H), 2.58 - 2.70 (m, 1 H), 2.73 - 2.89 (m, 1 H), 3.20 - 3.26 (m, 1 H), 3.37 - 3.45 (m, 1 H), 3.73 - 3.86 (m, 1 H), 3.88 - 3.97 (m, 1 H), 4.03 - 4.10 (m, 1 H), 4.11 - 4.18 (m, 1 H), 4.19 - 4.29 (m, 1 H), 4.29 - 4.37 (m, 1 H), 4.39 - 4.47 (m, 1 H), 4.57 - 4.60 (m, 2 H).

[0139] Example 2

[0140]

[0141] Synthesis route:

[0142]

[0143] Step 1: Synthesis of Compound 2-1

[0144] Compound 1-8 (630 mg, 1.18 mmol) was dissolved in dichloromethane (7 mL), and glacial acetic acid (85.23 mg, 1.42 mmol) and compound benzyl isocyanate (166.26 mg, 1.42 mmol) were added to the reaction system. The reaction was stirred at 20 °C for 16 h. The reaction was quenched with saturated ammonium chloride solution (20 mL) and extracted with dichloromethane (40 mL × 2). The organic phases were combined and washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (dichloromethane:methanol = 20:1) yielded compound 2-1. [M+1] + = 710.4.

[0145] Step 2: Synthesis of Compound 2-2

[0146] Compound 2-1 (519 mg, 731.12 μmol) was dissolved in anhydrous methanol (7.8 mL), and then a solution of potassium carbonate (252.61 mg, 1.83 mmol) in water (5.2 mL) was added to the reaction system. The reaction was stirred at 20 °C for 16 h. Water (10 mL) was added to the reaction mixture, and the mixture was extracted twice with dichloromethane (20 mL). The organic phase was washed with 3% citric acid (10 mL), then with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 2-2 was obtained without purification. [M+1]+ = 668.3.

[0147] Step 3: Synthesis of compounds 2-3

[0148] Compound 2-2 (420 mg, 628.90 μmol) was dissolved in dichloromethane (4.2 mL), and Dys-Martin periodide (346.76 mg, 817.57 μmol) was added to the reaction mixture. The mixture was stirred at 20 °C for 16 h. Sodium thiosulfate (15 mL) and saturated sodium bicarbonate solution (20 mL) were added to the reaction mixture, and the mixture was extracted twice with dichloromethane (50 mL). The combined organic phases were dried over anhydrous sulfuric acid, filtered, and concentrated. The solution was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to give compound 2-3. [M+1] + = 666.4.

[0149] Step 4: Synthesis of hydrochloride salts of compounds 2-4

[0150] Compound 2-3 (50 mg, 75.10 μmol) was dissolved in tetrahydrofuran (0.5 mL), and a 4 M hydrogen chloride solution in ethyl acetate (1.13 mL) was added to the reaction mixture. The reaction was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure and repeatedly rotary evaporated with a small amount of dichloromethane until a white foam was formed. The hydrochloride salt of compound 2-4 was obtained. [M+1] + = 566.4.

[0151] Step 5: Synthesis of Compound 2

[0152] The hydrochloride salts of compounds 2-4 (42.98 mg, 75.98 μmol) were dissolved in tetrahydrofuran (0.5 mL), cooled to 0 °C, and pyridine (42.07 mg, 531.83 μmol) and trifluoroacetic anhydride (63.83 mg, 303.91 μmol) were added. The reaction mixture was heated to room temperature (20 °C) and stirred for 16 h. The reaction solution was extracted with water (10 mL) and dichloromethane (10 mL × 2). The organic phases were combined and washed with 3% citric acid (10 mL), followed by washing with saturated brine (10 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 2 was obtained from the crude product by preparative HPLC. [M+1] + = 662.0.

[0153] Example 3

[0154]

[0155] Synthesis route:

[0156]

[0157] Step 1: Synthesis of Compound 3-1

[0158] Compound 1-8 (223 mg, 418.65 μmol) was dissolved in dichloromethane (2.5 mL), and glacial acetic acid (30.17 mg, 502.37 μmol) and tert-butyl isocyanate (41.76 mg, 502.37 μmol) were added to the reaction system. The reaction was stirred at 25 °C for 2 h. A saturated ammonium chloride solution (5 mL) was added to the reaction system and stirred for 10 min. Dichloromethane (10 mL) was added for extraction. The organic phase was washed with water (5 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Purification by silica gel column chromatography (dichloromethane:methanol = 10:1) yielded compound 3-1. [M+1] + = 676.4.

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

[0160] Compound 3-1 (122 mg, 180.51 μmol) was dissolved in methanol (2.5 mL), and then a solution of potassium carbonate (62.37 mg, 451.28 μmol) in water (1.5 mL) was added. The reaction was stirred at 20 °C for 16 h. 3% citric acid (10 mL) was added to the reaction mixture, and the mixture was extracted three times with dichloromethane (20 mL). The organic phase was washed with saturated sodium chloride solution (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound 3-2. [M+1] + = 634.4.

[0161] Step 3: Synthesis of compound 3-3

[0162] Compound 3-2 (1.2 g, 1.89 mmol) was dissolved in dichloromethane (15 mL), and then Dysmartin oxidant (1.04 g, 2.46 mmol) was added. The reaction was stirred at 20 °C for 18 h. Sodium thiosulfate (60 mL) and sodium bicarbonate solution (60 mL) were added to the reaction system, and the mixture was stirred for 10 min. Extraction was performed with dichloromethane (120 mL × 2). The organic phase was washed with saturated brine (60 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (DCM:MeOH = 20:1) yielded compound 3-3. [M+1] + = 632.5.

[0163] Step 4: Synthesis of compounds 3-4

[0164] Compound 3-3 (380 mg, 601.46 μmol) was dissolved in tetrahydrofuran (10 mL), and then ethyl acetate hydrochloride (4 M, 9.02 mL) was added. The reaction was stirred at 20 °C for 1 h. The reaction mixture was directly pumped to dryness and repeatedly concentrated under reduced pressure with a small amount of dichloromethane until white granules were obtained, yielding compound 3-4. [M+1]+ = 532.4.

[0165] Step 5: Synthesis of Compound 3

[0166] Compounds 3-4 (380 mg, 714.71 μmol) were dissolved in tetrahydrofuran (10 mL), and then trifluoroacetic anhydride (395.73 mg, 5 mmol) and pyridine (600.44 mg, 2.86 mmol) were added at 0 °C. The reaction mixture was stirred at 20 °C for 16 h. Water (60 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (120 mL × 2). The organic phase was washed successively with 3% citric acid (120 mL) and saturated sodium chloride solution (120 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 3 was obtained from the crude product by preparative HPLC. 1H NMR (400 MHz, CD3OD) δ ppm 0.88 - 1.33 (m, 6 H) 1.34- 1.45 (m, 9 H) 1.47 - 2.01 (m, 15 H) 2.19 - 2.44 (m, 1 H) 2.51 - 2.90 (m, 3H) 3.18 - 3.28 (m, 1 H) 3.69 - 4.03 (m, 2 H) 4.16 - 4.31 (m, 1 H) 4.37 - 4.48 (m, 1 H) 4.53 - 4.67 (m, 1 H). [M+1] + = 628.4.

[0167] Example 4

[0168]

[0169] Synthesis route:

[0170]

[0171] Step 1: Synthesis of compound 4-2

[0172] Compound 4-1 was dissolved in N,N-dimethylformamide (5 mL), and O-(7-azabenzotriazol-1-yl)- was added. N,N,N,N -Tetramethylurea hexafluorophosphate (1.24 g, 3.27 mmol) was stirred for 0.5 h, followed by the addition of diisopropylethylamine (1.41 g, 10.90 mmol, 1.90 mL) and compound 1-3 hydrochloride (527.06 mg, 2.40 mmol), and the reaction was carried out at 20 °C for 2 h. The reaction mixture was extracted with ethyl acetate (200 mL) and 3% citric acid solution (100 mL), and the organic phase was separated. The organic phase was then washed with semi-saturated brine (100 mL) until neutral, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:0~10:1) to obtain compound 4-2. 1H NMR (400MHz, CDCl3) δ = 5.44 -5.24 (m, 1H), 4.44 (dd, J=6.6, 8.8 Hz, 1H), 4.36 (d, J=3.8 Hz, 1H), 4.20 -4.14 (m, 2H), 3.85 (dd, J=7.9, 10.3 Hz, 1H), 3.66 (dd, J=3.5, 10.3 Hz, 1H), 2.75 - 2.64 (m, 2H), 1.98 - 1.59 (m, 13H), 1.50 - 1.43 (m, 9H), 1.27 - 1.25(m, 3H).

[0173] Step 2: Synthesis of compound 4-3

[0174] 4-2 (0.8 g, 2.03 mmol) was dissolved in tetrahydrofuran (10 mL) and water (6 mL), and lithium hydroxide monohydrate (170.19 mg, 4.06 mmol) was added. The mixture was stirred at 20 °C for 16 h. The reaction system was extracted with dichloromethane (200 mL) and 3% citric acid (100 mL), and the organic phase was separated. The organic phase was washed with saturated brine (100 mL) until neutral, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 4-3. 1 H NMR (400MHz, CD3OD) δ = 4.36 (br d, J =8.3 Hz, 1H), 4.29 -4.23 (m, 1H), 3.95 - 3.77 (m, 2H), 3.73 (br t, J =6.2 Hz, 1H), 2.76 - 2.61 (m,2H), 2.02 - 1.57 (m, 13H), 1.44 (s, 9H).

[0175] Step 3: Synthesis of compound 4-4

[0176] 4-3 (0.7 g, 1.91 mmol) was dissolved in 2-butanone (15 mL), and 1-hydroxybenzotriazole (258.11 mg, 1.91 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (439.43 mg, 2.29 mmol) were added. The mixture was stirred for 0.5 h, followed by the addition of diisopropylethylamine (1.23 g, 9.55 mmol, 1.66 mL) and compound 1-2 hydrochloride (467.88 mg, 2.10 mmol). The reaction mixture was then reacted at 20 °C for 16 h. The reaction mixture was extracted with 3% citric acid (100 mL) and dichloromethane (200 mL) to obtain the organic phase. The organic phase was then extracted again with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Compound 4-4 was obtained by purification by silica gel column chromatography (dichloromethane:methanol = 1:0~10:1). 1 H NMR (400MHz, CD3OD) δ = 8.65 (br d, J =8.4 Hz, 1H), 7.99 - 7.69 (m, 1H), 7.61 -7.44 (m, 1H), 4.58 - 4.44 (m, 1H), 4.34 (d, J =8.0 Hz, 1H), 4.22 (d, J =4.0 Hz, 1H), 3.92 (dd, J =7.9, 10.3 Hz, 1H), 3.78 (br dd, J =3.6, 10.3 Hz, 1H), 3.72 (s,3H), 3.30 - 3.26 (m, 2H), 2.89 - 2.77 (m, 1H), 2.75 - 2.50 (m, 3H), 2.36 -2.25 (m, 1H), 2.21 - 2.08 (m, 1H), 2.05 - 1.52 (m, 14H), 1.49 - 1.39 (m, 9H).

[0177] Step 4: Synthesis of compounds 4-5

[0178] Compound 4-4 (0.75 g, 1.40 mmol) was dissolved in tetrahydrofuran (7.5 mL), cooled to 0 °C, and lithium borohydride (91.66 mg, 4.21 mmol) was added. The mixture was slowly heated to 20 °C and reacted for 1 h. Semi-saturated NH4Cl (30 mL) was added to the reaction system and stirred for 30 min to quench the reaction. Dichloromethane (60 mL × 2) was then added for extraction, and the organic phase was separated, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 4-5. 1 H NMR (400MHz, CD3OD) δ = 4.35 (br d, J =7.9 Hz,1H), 4.22 - 4.14 (m, 1H), 4.04 - 3.89 (m, 2H), 3.85 - 3.66 (m, 2H), 3.50 (brt, J =5.2 Hz, 2H), 3.29 - 3.25 (m, 1H), 2.91 - 2.78 (m, 1H), 2.74 - 2.56 (m,3H), 2.44 (br d, J =7.6 Hz, 1H), 2.36 - 2.25 (m, 1H), 2.00 - 1.74 (m, 14H), 1.44 (s, 9H).

[0179] Step 5: Synthesis of compounds 4-6

[0180] Compounds 4-5 (0.55 g, 1.09 mmol) were dissolved in dichloromethane (20 mL), and Dys-Martin oxidant (506.49 mg, 1.19 mmol) was added. The mixture was stirred at 25 °C for 16 h. The reaction mixture was then extracted with saturated sodium thiosulfate solution (30 mL × 2) and saturated sodium bicarbonate solution (30 mL × 2) and dichloromethane (60 mL). The organic phase was separated, dried over anhydrous sodium sulfate, filtered, and concentrated to give compounds 4-6. 1 H NMR (400MHz, CDCl3) δ = 9.56 - 9.44 (m, 1H),8.01 (br d, J =6.0 Hz, 1H), 6.14 - 5.97 (m, 1H), 5.50 - 5.35 (m, 1H), 4.55 -4.29 (m, 3H), 3.94 - 3.83 (m, 1H), 3.65 (br d, J=7.3 Hz, 1H), 3.42 - 3.29 (m,2H), 2.94 - 2.76 (m, 2H), 2.74 - 2.68 (m, 1H), 2.60 - 2.49 (m, 1H), 2.39 (td, J =3.2, 5.9 Hz, 1H), 1.97 - 1.60 (m, 15H), 1.44 (s, 8H).

[0181] Step 6: Synthesis of compounds 4-7

[0182] Compounds 4-6 (0.75 g, 1.49 mmol) were dissolved in dichloromethane (7.5 mL), and glacial acetic acid (107.10 mg, 1.78 mmol) and tert-butyl isocyanate (148.27 mg, 1.78 mmol) were added. The mixture was stirred at 20 °C for 2 h. The reaction mixture was extracted with saturated ammonium chloride solution (30 mL) and dichloromethane (60 mL), and the organic phase was separated, dried over anhydrous sodium sulfate, filtered, and concentrated to give compounds 4-7. [M+1] + =648.2.

[0183] Step 7: Synthesis of compounds 4-8

[0184] Compound 4-7 (0.8 g, 1.23 mmol) was dissolved in methanol (6.5 mL) and water (4 mL), and potassium carbonate (426.69 mg, 3.09 mmol) was added. The mixture was stirred at 20 °C for 2 h. A saturated sodium bicarbonate solution (30 mL) was added to the reaction mixture, and the mixture was stirred for 10 min. Dichloromethane (60 mL) was added for extraction, and the organic phase was separated, dried over anhydrous sodium sulfate, filtered, and concentrated. The solution was purified by silica gel column chromatography (dichloromethane:methanol = 1:0–10:1) to give compound 4-8. 1 H NMR (400MHz, CDCl3) δ = 7.08(br d, J =8.3 Hz, 1H), 6.87 - 6.62 (m, 1H), 6.07 - 5.94 (m, 1H), 5.98 (br s,1H), 5.35 (br d, J =8.6 Hz, 1H), 4.52 - 4.30 (m, 2H), 4.28 - 3.98 (m, 3H), 3.74- 3.57 (m, 1H), 3.33 (br d, J =8.3 Hz, 2H), 2.81 - 2.65 (m, 3H), 2.54 (td, J=8.0, 15.5 Hz, 1H), 2.41 - 2.31 (m, 1H), 2.03 - 1.69 (m, 13H), 1.63 - 1.52 (m,2H), 1.46 - 1.41 (m, 9H), 1.38 - 1.32 (m, 9H).

[0185] Step 8: Synthesis of compounds 4-9

[0186] Compounds 4-8 (0.65 g, 1.07 mmol) were dissolved in dichloromethane (12 mL), and Dys-Martin oxidant (682.67 mg, 1.61 mmol) was added. The reaction mixture was reacted at 20 °C for 16 h. The reaction mixture was then extracted with saturated sodium thiosulfate solution (30 mL × 2) and saturated sodium bicarbonate solution (30 mL × 2) and dichloromethane (60 mL). The organic phase was separated, dried over anhydrous sodium sulfate, filtered, and concentrated. Compounds 4-9 were purified by silica gel column chromatography (dichloromethane:methanol = 1:0–10:1). [M+1] + =604.5.

[0187] Step 9: Synthesis of trifluoroacetates of compounds 4-10

[0188] Compound 4-9 (450 mg, 745.34 μmol) was dissolved in dichloromethane (6 mL), and trifluoroacetic acid (1.5 mL) was added. The mixture was reacted at 20 °C for 1 h. The reaction system was concentrated under reduced pressure to give the trifluoroacetate of the target compound 4-10. 1 H NMR (400MHz, DMSO-) d6 ) δ = 8.51 (d, J =8.4 Hz, 1H), 8.04 (br s, 3H), 7.64 (s, 1H), 5.75 (s,1H), 5.23 - 5.05 (m, 1H), 4.29 - 4.22 (m, 1H), 4.20 - 4.10 (m, 1H), 3.79 -3.65 (m, 1H), 3.63 - 3.55 (m, 1H), 3.15 - 3.04 (m, 1H), 2.69 (br d, J =7.4 Hz,3H), 2.44 - 2.34 (m, 1H), 2.25 - 2.17 (m, 1H), 1.85 - 1.51 (m, 13H), 1.48 -1.33 (m, 2H), 1.30 (s, 9H).

[0189] Step 10: Synthesis of Compound 4

[0190] 4–10% trifluoroacetate (275 mg, 546.03 μmol) was dissolved in dichloromethane (2.7 mL). Pyridine (431.91 mg, 5.46 mmol) and trifluoroacetic anhydride (286.71 mg, 1.37 mmol) were added at 0 °C, and the reaction was carried out at 20 °C for 2 h. The reaction mixture was extracted with 3% citric acid solution (30 mL) and dichloromethane (60 mL x 2), and the organic phase was separated. The mixture was washed with saturated brine (60 mL) until neutral, dried over anhydrous sodium sulfate, filtered, and concentrated. The solution was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to give compound 4. 1 H NMR (400MHz, CDCl3) δ = 8.09 (br d, J =5.9 Hz, 1H), 7.40 (brd, J =8.6 Hz, 1H), 6.89 - 6.71 (m, 1H), 5.92 - 5.79 (m, 1H), 5.41 - 5.20 (m,1H), 4.90 - 4.68 (m, 1H), 4.28 (d, J =3.5 Hz, 1H), 4.01 - 3.82 (m, 1H), 3.61 -3.46 (m, 1H), 3.40 - 3.29 (m, 2H), 2.86 (br d, J =11.0 Hz, 1H), 2.81 - 2.74 (m,1H), 2.55 - 2.42 (m, 1H), 2.00 - 1.62 (m, 17H), 1.40 - 1.37 (m, 9H).

[0191] Example 5

[0192]

[0193] Synthesis route:

[0194]

[0195] Step 1: Synthesis of Compound 5-2

[0196] Compound 5-1 (0.65 g, 2.67 mmol) was dissolved in N,N-dimethylformamide (20 mL), and then O-(7-azabenzotriazol-1-yl)- N,N,N,N Tetramethylurea hexafluorophosphate (1.22 g, 3.21 mmol) was added to the reaction system, and the mixture was stirred at 20 °C for 0.5 h. Then, diisopropylethylamine (863.20 mg, 6.68 mmol) and compound 1-3 hydrochloride (704.37 mg, 3.21 mmol) were added separately to the reaction system, and the reaction was stirred at 20 °C for 5 h. The reaction mixture was directly diluted with 3% citric acid (30 mL), extracted with ethyl acetate (60 mL), washed with saturated brine (30 mL), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The solution was purified by column chromatography, and the fraction was directly concentrated under reduced pressure to give compound 5-2. [M+1] + =409.2.

[0197] Step 2: Synthesis of Compound 5-3

[0198] Compound 5-2 (1 g, 2.11 mmol) was dissolved in tetrahydrofuran (18 mL) and water (6 mL). Lithium hydroxide monohydrate (176.83 mg, 4.21 mmol) was added to the reaction mixture. The reaction was stirred at 25 °C for 12 h. The reaction mixture was diluted with 3% citric acid (30 mL) and extracted with ethyl acetate (60 mL). The organic phases were combined and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The product required no purification. Compound 5-3 was obtained. [M+1] + =381.2.

[0199] Step 3: Synthesis of Compounds 5-4

[0200] Compound 5-3 (1 g, 2.63 mmol) was dissolved in 2-butanone (40 mL), then 1-hydroxybenzotriazole (355.13 mg, 2.63 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (604.61 mg, 3.15 mmol) were added to the reaction system, and the reaction was stirred at 25 °C for 0.5 h. Then, diisopropylethylamine (1.36 g, 10.51 mmol) and compound 1-2 hydrochloride (702.28 mg, 3.15 mmol) were added to the reaction system, and the reaction was stirred for another 16 h. The reaction mixture was extracted with dichloromethane (30 mL), washed with 3% citric acid (30 mL), washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by column chromatography yielded compound 5-4. [M+1] + = 549.3.

[0201] Step 4: Synthesis of compound 5-5

[0202] Compound 5-4 (1.24 g, 2.26 mmol) was dissolved in tetrahydrofuran (30 mL), and then lithium borohydride (147.67 mg, 6.78 mmol) was added to the reaction system. The reaction was stirred at 25 °C for 3 h. The reaction mixture was extracted with dichloromethane (30 mL), the organic phase was quenched with saturated ammonium chloride (30 mL), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 5-5. [M+1] + = 521.3.

[0203] Step 5: Synthesis of compounds 5-6

[0204] Compound 5-5 (0.5 g, 960.32 μmol) was dissolved in dichloromethane (20 mL), and then Dysmart reagent (448.04 mg, 1.06 mmol) was added to the reaction system. The reaction was stirred at 25 °C for 16 h. The reaction was quenched with saturated sodium bicarbonate solution (20 mL) and sodium thiosulfate solution (20 mL), and extracted with dichloromethane (60 mL). The solution was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 5-6. [M+1] + = 519.3.

[0205] Step 6: Synthesis of compounds 5-7

[0206] Compounds 5-6 (0.6 g, 1.16 mmol) and glacial acetic acid (83.36 mg, 1.39 mmol) were dissolved in dichloromethane (20 mL), and then tert-butyl isocyanate (115.40 mg, 1.39 mmol) was added to the reaction mixture. The reaction mixture was stirred at 25 °C for 3 h. The reaction mixture was directly diluted with water (20 mL), extracted with dichloromethane (60 mL), and the organic phases were combined, dried, and concentrated. The mixture was purified by column chromatography to give compound 5-7. [M+1] + = 662.4.

[0207] Step 7: Synthesis of compounds 5-8

[0208] Compound 5-7 (0.4 g, 604.39 μmol) was dissolved in methanol (12 mL), and then an aqueous solution of potassium carbonate (208.83 mg, 1.51 mmol) (6 mL) was added to the reaction system. The reaction mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with water (50 mL), extracted with dichloromethane (60 mL), and the organic phases were combined and washed with 3% citric acid (15 mL) and then with saturated brine (20 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 5-8. [M+1] + = 620.4.

[0209] Step 8: Synthesis of compounds 5-9

[0210] Compound 5-8 (0.35 g, 564.71 μmol) was dissolved in dichloromethane (10 mL), and then Dys-Martin reagent (359.27 mg, 847.06 μmol) was added to the reaction system. The reaction was stirred at 25 °C for 12 h. The reaction was quenched with saturated sodium bicarbonate solution (20 mL) and sodium thiosulfate solution (20 mL), extracted with dichloromethane (20 mL × 3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated to give compound 5-9. [M+1] + = 618.4.

[0211] Step 9: Synthesis of compounds 5-10

[0212] Compound 5-9 (240 mg, 388.49 μmol) was dissolved in dichloromethane (6 mL), and trifluoroacetic acid (1.5 mL) was added. The reaction was carried out at 20 °C for 1 h. The reaction mixture was concentrated to give compound 5-10. [M+1] + =518.4.

[0213] Step 10: Synthesis of Compound 5

[0214] Compound 5-10 (180 mg, 347.72 μmol) was dissolved in dichloromethane (3 mL). Pyridine (275.05 mg, 3.48 mmol) and trifluoroacetic anhydride (182.58 mg, 869.30 μmol) were added at 0 °C, and the reaction was carried out at 20 °C for 2 h. The reaction mixture was then extracted with dichloromethane (60 mL) and 3% citric acid solution (30 mL) to obtain the organic phase. This phase was further extracted with saturated brine (30 mL), dried over anhydrous sodium sulfate, and concentrated. Compound 5 was purified by column chromatography. 1 H NMR (400 MHz, DMSO-) d6) δ ppm 9.79 - 9.92 (m, 1 H) 8.46 (d, J=8.13 Hz, 1 H) 7.12 -7.99 (m, 2 H) 5.95 - 6.29 (m, 1 H) 4.89 - 5.14 (m, 1 H) 4.02 - 4.38 (m, 2 H)3.57 - 3.86 (m, 2 H) 2.87 - 3.30 (m, 3 H) 2.63 - 2.76 (m, 1 H) 2.01 - 2.42(m, 3 H) 1.36 - 1.99 (m, 15 H) 1.26 - 1.33 (m, 9 H) 1.15 - 1.26 (m, 2 H).

[0215] Example 6

[0216]

[0217] Synthesis route:

[0218]

[0219] Step 1: Synthesis of Compound 6-2

[0220] Sodium borohydride (10.42 g, 275.44 mmol) was dissolved in tetrahydrofuran (300 mL), purged three times with nitrogen, and cooled to 0 °C. Compound 6-1 (20 g, 140.65 mmol) was dissolved in tetrahydrofuran (100 mL) and slowly added dropwise to the system, followed by slow dropwise addition of boron trifluoride diethyl ether (281.30 mmol, 34.72 mL). The reaction mixture was heated to 20 °C and stirred for 2 h. Ethanol (1 L) was added, and the mixture was stirred for 15 min. The reaction mixture was concentrated, and water (100 mL) was added. The mixture was extracted with dichloromethane (100 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 6-2. 1 H NMR (400 MHz, CDCl3) δ ppm 3.6 (s, 2 H), 1.40 - 1.56 (m, 5 H), 1.23 - 1.37 (m, 5 H), 0.93 (s, 3 H).

[0221] Step 2: Synthesis of Compound 6-3

[0222] Compound 6-2 (17.99 g, 140.32 mmol) was dissolved in dichloromethane (180 mL), and pyridine chlorochromate (45.37 g, 210.47 mmol) and silica gel (45 g, 748.59 mmol) were added. The reaction mixture was stirred at 20 °C for 16 h. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure at 20 °C to obtain crude product 6-3, which was directly used in the next reaction step. 1 H NMR (400 MHz, CDCl3) δ ppm 9.37 (s, 1 H), 1.16 - 1.54 (m, 10 H), 0.93 (s, 3 H).

[0223] Step 3: Synthesis of Compound 6-4

[0224] Compound 6-3 (17 g, 134.71 mmol) was dissolved in chloroform (150 mL), and R-phenylglycine (18.48 g, 134.71 mmol) was added. The reaction mixture was stirred at 20 °C for 2 h, cooled to 0 °C, and trimethylcyanosilane (26.73 g, 269.42 mmol, 33.71 mL) was added. The reaction mixture was stirred at 20 °C for 16 h. The reaction mixture was concentrated to dryness under reduced pressure. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1) to give compound 6-4. 1 H NMR (400 MHz, CDCl3) δppm 7.13- 7.27 (m, 5 H), 3.89 - 3.96 (m, 1 H), 3.61 - 3.69 (m, 1 H), 3.37 -3.47 (m, 1 H), 2.92 - 3.02 (m, 1 H), 1.03- 1.41 (m, 10 H), 0.93 (s, 3 H). [M+1] + = 273.2.

[0225] Step 4: Synthesis of compounds 6-5

[0226] Compound 6-4 (10 g, 36.71 mmol) was dissolved in methanol (100 mL) and dichloromethane (100 mL), cooled to 0 °C, and lead tetraacetate (13.56 g, 27.53 mmol) was added. Nitrogen gas was purged three times, and the reaction was stirred at 0 °C for 2 h. A saturated sodium bicarbonate solution (200 mL) was added, and the mixture was extracted with dichloromethane (45 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude 6-5. [M+1] + = 241.0.

[0227] Step 5: Synthesis of Compound 6-6

[0228] Compound 6-5 (5.4 g, 22.47 mmol) was dissolved in hydrochloric acid (6 M, 490 mL), and the reaction solution was heated to 100 °C and stirred for 24 h. The reaction solution was cooled to room temperature, extracted with chloroform (300 mL × 3), and concentrated under reduced pressure in aqueous phase to give compound 6-6. 1 H NMR (400 MHz, CD3OD) δ ppm 4.46 (s, 1 H), 1.41- 1.74 (m, 10 H), 1.20 (s, 3H).

[0229] Step 6: Synthesis of compounds 6-7

[0230] Compound 6-6 (3.6 g, 17.33 mmol) was dissolved in methanol (36 mL), and triethylamine (52.00 mmol, 7.24 mL) and di-tert-butyl dicarbonate (26.00 mmol, 5.97 mL) were added. The reaction mixture was stirred at 20 °C for 4 h. The pH of the reaction mixture was adjusted to 3 with citric acid aqueous solution (3%), and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 6-7.

[0231] Step 7: Synthesis of compounds 6-8

[0232] Compounds 6-7 (1.5 g, 5.53 mmol) and the hydrochloride salts of compounds 1-3 (1.46 g, 6.63 mmol) were dissolved in ethyl acetate (15 mL), and then... N,N -Diisopropylethylamine (2.86 g, 22.11 mmol, 3.85 mL) and n-propylphosphoric anhydride (5.28 g, 8.29 mmol, 4.93 mL, 50% ethyl acetate solution) were reacted and stirred at 55 °C for 16 h. Water (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 100:0 ~ 10:1) to give compounds 6-8. [M+1] + = 437.1.

[0233] Step 8: Synthesis of compounds 6-9

[0234] Compounds 6-8 (1.52 g, 3.48 mmol) were dissolved in methanol (15 mL) and water (5 mL), and lithium hydroxide monohydrate (166.77 mg, 6.96 mmol) was added. The reaction mixture was stirred at 20 °C for 16 h. The pH of the reaction mixture was adjusted to 3 with 3% citric acid aqueous solution, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compounds 6-9. [M-56+H] + = 353.2.

[0235] Step 9: Synthesis of compounds 6-10

[0236] Compounds 6-9 (1.42 g, 3.48 mmol) and 1-1 (1.50 g, 4.18 mmol, TsOH) were dissolved in ethyl acetate (15 mL), and then... N,N -Diisopropylethylamine (1.80 g, 13.92 mmol, 2.42 mL) and n-propylphosphoric anhydride (5.28 g, 8.29 mmol, 4.93 mL, 50% ethyl acetate solution) were reacted at 55 °C with stirring for 16 h. 30 mL of water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. Purification by silica gel column chromatography (dichloromethane:methanol = 100:0 - 10:1) gave compound 6-10. [M+1] + = 577.4.

[0237] Step 10: Synthesis of Compounds 6-11

[0238] Compound 6-10 (605 mg, 1.05 mmol) was dissolved in THF (12 mL), and LiBH4 (45 mg, 2.1 mmol) was slowly added at 0 °C. The mixture was then slowly heated to 20 °C and reacted for 2 h. A saturated ammonium chloride solution (10 mL) was slowly added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 6-11. [M+1] + = 549.1.

[0239] Step 11: Synthesis of compounds 6-12

[0240] Compound 6-11 (575 mg, 1.05 mmol) was dissolved in dichloromethane (10 mL), and Dysmart reagent (533.35 mg, 1.26 mmol) was added. The reaction mixture was stirred at 20 °C for 16 h. Saturated sodium thiosulfate solution (10 mL) and saturated sodium bicarbonate solution (10 mL) were added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 100:0-10:1) to give compound 6-12. [M+1] + = 547.1.

[0241] Step 12: Synthesis of compounds 6-13

[0242] Compound 6-12 (330 mg, 603.63 μmol) was dissolved in dichloromethane (3 mL), and acetic acid (72.50 mg, 1.21 mmol, 69.04 μL) and cyclopentyl isocyanate (68.92 mg, 724.35 μmol, 80.14 μL) were added. The reaction mixture was stirred at 20 °C for 2 h. A saturated ammonium chloride aqueous solution (5 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (10 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 100:0 to 10:1) to give compound 6-13. [M+1] + = 702.4.

[0243] Step 13: Synthesis of compounds 6-14

[0244] Compound 6-13 (304 mg, 433.12 μmol) was dissolved in methanol (3 mL) and water (2 mL), and potassium carbonate (149.65 mg, 1.08 mmol) was added. The reaction mixture was stirred at 20°C for 4 h. The pH of the reaction mixture was adjusted to 3 with 3% citric acid aqueous solution, and the mixture was extracted with ethyl acetate (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude compound 6-14, which was used directly in the next reaction. [M+1] + = 660.2.

[0245] Step 14: Synthesis of compounds 6-15

[0246] Compound 6-14 (285.7 mg, 432.97 μmol) was dissolved in dichloromethane (6 mL), and Dysmart reagent (220.37 mg, 519.57 μmol) was added. The reaction mixture was stirred at 20°C for 16 h. Saturated sodium thiosulfate solution (10 mL) and saturated sodium bicarbonate solution (10 mL) were added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 100:0 to 10:1) to give 6-15. [M+1] + = 658.4.

[0247] Step 15: Synthesis of trifluoroacetate of compound 6-16

[0248] Compound 6-15 (265.8 mg, 404.05 μmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (921.40 mg, 8.08 mmol, 598.31 μL) was added. The reaction mixture was stirred at 20 °C for 2 h. The reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate of compound 6-16, which was used directly in the next reaction. [M+1] + =558.3.

[0249] Step 16: Synthesis of Compound 6

[0250] The trifluoroacetate of compound 6-16 (271 mg, 403.43 μmol) was dissolved in dichloromethane (3 mL). Pyridine (223.38 mg, 2.82 mmol, 227.94 μL) and trifluoroacetic anhydride (127.10 mg, 605.14 μmol, 84.17 μL) were added at 0 °C. The reaction mixture was slowly heated to 20 °C and stirred for 2 h. Water (10 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined, washed with 3% citric acid (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Xtimate C18 100*30mm*3μm; mobile phase: [A: water (formic acid)-B: acetonitrile]; acetonitrile%: 40%-80%, 8min) to obtain compound 6. 1H NMR(400 MHz, CDCl3) δ ppm 7.01 - 7.27 (m, 1 H), 6.76 - 6.99 (m, 1 H), 6.05 -6.46 (m, 1 H), 5.02 - 5.43 (m, 1 H), 4.59 - 4.75 (m, 1 H), 4.25 - 4.49 (m, 1H), 4.09 - 4.23 (m, 1 H), 3.75-4.08 (m, 1 H), 3.59 - 3.73 (m, 1 H), 3.25 -3.54(m, 2 H), 2.42 - 2.92 (m, 3 H), 1.82-2.37 (m, 8 H), 1.25 - 1.80 (m, 20H), 0.90-1.20 (m, 4 H). [M+1] + = 654.4.

[0251] Example 7

[0252]

[0253] Synthesis route:

[0254]

[0255]

[0256] Step 1: Synthesis of Compound 7-2

[0257] 7-1 (90 g, 348.41 mmol) was dissolved in dichloromethane (900 mL), and Dysmart oxidant (162.55 g, 383.26 mmol) was added. The reaction mixture was stirred at 20 °C for 2 h. The reaction mixture was quenched with a saturated sodium thiosulfate aqueous solution (900 mL), and the pH was adjusted to 7-8 with a sodium carbonate aqueous solution. The mixture was separated, and the aqueous phase was extracted with dichloromethane (900 mL). The organic phases were combined and concentrated under reduced pressure to give compound 7-2. The crude product was used directly in the next reaction.

[0258] Step 2: Synthesis of Compound 7-3

[0259] N-cyclopentylformamide (2.79 g, 24.68 mmol) was dissolved in dichloromethane (55 mL), and Burgess reagent (7.67 g, 32.19 mmol) was added at 20 °C. The reaction mixture was stirred at 20 °C for 1 hour. Water (1.93 mL) was added and the mixture was stirred for 30 minutes. Then 7-2 (5.5 g, 21.46 mmol) and acetic acid (2.45 mL) were added, and the reaction mixture was stirred at 20 °C for 12 hours. A 2.5% sodium hypochlorite aqueous solution (105.62 mL) was added to the reaction mixture, and the mixture was stirred for 30 minutes. The reaction mixture was separated, and the aqueous phase was extracted with dichloromethane (50 mL × 2). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the crude product 7-3 was used directly in the next step.

[0260] Step 3: Synthesis of Compound 7-4

[0261] 7-3 (2 g, 5.41 mmol) was dissolved in methanol (60 mL) and water (30 mL), and potassium carbonate (5.04 g, 36.45 mmol) was added. The reaction mixture was stirred at 20 °C for 2 hours. The pH of the reaction mixture was adjusted to 5-6 with saturated citric acid solution, and then extracted with dichloromethane (100 mL). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was added to ethyl acetate (50 mL), and n-heptane (50 mL) was added dropwise with stirring. The mixture was stirred for 2 hours, the solid was filtered, and the filter cake was dried under vacuum to give compound 7-4.

[0262] [M-100+1] + = 269.9.

[0263] Step 4: Synthesis of hydrochloride salts of compounds 7-5

[0264] 7-4 (10 g, 24.30 mmol) was dissolved in ethyl acetate (20 mL), and ethyl acetate hydrochloride solution (4 M, 5.41 mL) was added. The reaction mixture was stirred at 25 °C for 12 hours. The reaction mixture was filtered, and the filtrate was dried under vacuum to give the hydrochloride salt of compound 7-5. [M+1] + = 270.0.

[0265] Step 5: Synthesis of Compound 7-7

[0266] Compound 7-6 (2.00 g, 6.04 mmol) was dissolved in tetrahydrofuran (30 mL), and zinc powder (3.25 g, 49.70 mmol), zirconium dichloroethylene (2.19 g, 7.24 mmol), and dibromomethane (1.15 g, 6.64 mmol) were added. The reaction mixture was heated to 80 °C and stirred for 5 hours. After the reaction was complete, the mixture was cooled to room temperature, water (5 mL) was added, and the mixture was filtered. The filtrate was collected and extracted with methyl tert-butyl ether (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by rapid silica gel chromatography (ISCO®; 12 g SepaFlash® rapid silica gel column, eluent 0–20% ethyl acetate / petroleum ether, flow rate 30 mL / min) to obtain compound 7-7. [M+1] + = 330.1.

[0267] Step 6: Synthesis of compounds 7-8

[0268] Under nitrogen protection, diethylzinc (1 M, THF, 37.95 mL) was added to dichloromethane (45 mL), and the reaction mixture was stirred at 0°C. Then, trifluoroacetic acid (4.33 g, 37.95 mmol) was added, and the mixture was stirred for 30 minutes. A dichloromethane solution of compound 7-7 (2.50 g, 7.59 mmol) was added (70 mL), and the reaction mixture was slowly heated to 25°C and stirred for 16 hours. A saturated ammonium chloride aqueous solution (100 mL) was added, and the mixture was separated. The aqueous phase was extracted with dichloromethane (100 mL × 3), and the organic phases were combined and concentrated under reduced pressure. The crude product was purified to compound 7-8 by rapid silica gel chromatography (ISCO®; 12 g SepaFlash® rapid silica gel column, eluent 0-20% ethyl acetate / petroleum ether, flow rate 30 mL / min). [M+1] + = 344.1.

[0269] Step 7: Synthesis of compounds 7-9

[0270] Compounds 7-8 (600.00 mg, 1.75 mmol) were dissolved in ethanol (30 mL), and dioxane hydrochloride solution (4 M, 500.00 μL) and dry palladium on carbon (200 mg, 10%) were added. The reaction mixture was stirred at 25 °C for 12 hours under hydrogen (15 Psi). The reaction mixture was filtered, and the filtrate was collected and concentrated under reduced pressure to obtain compounds 7-9. The crude product was used directly in the next step.

[0271] Step 8: Synthesis of compounds 7-10

[0272] Compounds 7-9A (852.41 mg, 3.69 mmol) and 7-9 (735.00 mg, 3.51 mmol) were dissolved in N,N-dimethylformamide (30 mL) and dichloromethane (40 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (1.81 g, 4.77 mmol) and N,N-diisopropylethylamine (1.36 g, 10.53 mmol) were added. The reaction mixture was stirred at 25°C for 16 hours under nitrogen protection. The reaction mixture was then poured into 200 mL of 4M citric acid aqueous solution, extracted with dichloromethane (100 mL × 3), and the organic phases were combined and concentrated under reduced pressure. The crude product was purified by rapid silica gel chromatography (ISCO®; 12g SepaFlash® rapid silica gel column, eluent 0-20% ethyl acetate / petroleum ether, flow rate 30 mL / min) to give compound 7-10. [M+Na] + = 445.2

[0273] Step 9: Synthesis of Compounds 7-11

[0274] Compound 7-10 (1.30 g, 3.08 mmol) was dissolved in tetrahydrofuran (6 mL), water (6 mL), and methanol (6 mL). Lithium hydroxide monohydrate (387.30 mg, 9.23 mmol) was added, and the reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was diluted in 100 mL of dichloromethane, and then 50 mL of 1N hydrochloric acid aqueous solution was added. The organic phase was extracted separately, and the aqueous phase was extracted with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound 7-11. The crude product was used directly in the next step. [M-tBu+1] + = 339.1.

[0275] Step 10: Synthesis of the hydrochloride salt of compound 7-12

[0276] Compound 7-11 (1.20 g, 3.04 mmol) was dissolved in tert-butyl methyl ether (10 mL), and ethyl acetate hydrochloride (4 M, 760.45 μL) was added. The reaction mixture was stirred at 25 °C for 0.5 h. The reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of compound 7-12, and the crude product was used directly in the next step. [M+1] + = 295.1.

[0277] Step 11: Synthesis of compounds 7-13

[0278] The hydrochloride salt of compound 7-12 (1.20 g, 3.63 mmol) was dissolved in methanol (10 mL), and triethylamine (1.84 g, 18.14 mmol) and ethyl trifluoroacetate (1.03 g, 7.25 mmol) were added. The reaction mixture was stirred at 25 °C for 12 hours. The reaction mixture was concentrated under reduced pressure, and dichloromethane (100 mL) was added. The mixture was washed with 100 mL of 5% citric acid aqueous solution, and the liquid-liquid extraction was performed. The aqueous phase was extracted with dichloromethane (200 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate and concentrated to dryness. The crude product was stirred at 25 °C for 1 hour with a mixed solvent of methyl tert-butyl ether and petroleum ether (1:6, 10 mL), and filtered to obtain crude product 7-13, which was used directly in the next step. [M +1] + = 390.9.

[0279] Step 12: Synthesis of compounds 7-13A and 7-13B

[0280] Compound 7-13 (100 mg, 256.15 μmol) was purified by preparative HPLC (column: Xtimate C18150*40mm*5μm; mobile phase: [A: water (hydrochloric acid)-B: acetonitrile]; acetonitrile%: 43%-63%, 10 min) to obtain 7-13A (first peak, retention time: 4.198 min, [M+1]). + = 391.2) and 7-13B (later peak, retention time: 4.377 min, [M+1]) + =391.2). Analytical method: Column: ChromCore 120 C18 3μm, 3.0*30mm; Mobile phase: [A: water (trifluoroacetic acid) - B: acetonitrile]; Acetonitrile%: 10%-80% over 6 minutes, then 80% for 0.5 minutes, flow rate 0.8 mL / min.

[0281] Step 13: Synthesis of compound 7-14A

[0282] Compound 7-13A (35 mg, 89.65 μmol) and the hydrochloride of 7-5 (41.12 mg) were dissolved in DCM (0.5 mL). Tris(1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride) (20.62 mg, 107.58 μmol), 2-hydroxypyridine-1-oxide (2.49 mg, 22.41 μmol), and N,N-diisopropylethylamine (34.76 mg, 268.96 μmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was diluted with dichloromethane (5 mL), washed with saturated citric acid aqueous solution (1 mL) and saturated brine (1 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated to obtain crude product 7-14A, which was used directly in the next step. [M +1] + = 642.2

[0283] Step 14: Synthesis of compound 7-14B

[0284] Compounds 7-13B (25.00 mg, 64.04 μmol) and 7-5 hydrochloride (29.37 mg) were dissolved in DCM (0.5 mL). Tris(1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride) (14.73 mg, 76.84 μmol), 2-hydroxypyridine-1-oxide (1.78 mg, 16.01 μmol), and N,N-diisopropylethylamine (24.83 mg, 192.11 μmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was diluted with dichloromethane (5 mL), washed with saturated citric acid aqueous solution (1 mL) and saturated brine (1 mL), dried over anhydrous sodium sulfate, and the organic phase was concentrated to dryness to obtain crude 7-14B, which was used directly in the next step. [M +1] + = 642.2

[0285] Step 15: Synthesis of Compound 7A

[0286] Compound 7-14A (50 mg, 77.92 μmol) was dissolved in DCM (0.5 mL), and Dysmartin oxidant (39.66 mg, 93.50 μmol) was added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was diluted with dichloromethane (10 mL) and washed with saturated sodium thiosulfate aqueous solution (2 mL), saturated sodium bicarbonate aqueous solution (2 mL), and saturated brine (2 mL), respectively. The solution was dried over anhydrous sodium sulfate, and the organic phase was concentrated to dryness. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1 to 0:1) to give compound 7A. [M +1] + = 640.2.

[0287] Step 16: Synthesis of Compound 7B

[0288] Compound 7-14B (35 mg, 54.54 μmol) was dissolved in DCM (0.5 mL), and Dysmartin oxidant (27.76 mg, 65.45 μmol) was added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was diluted with dichloromethane (10 mL) and washed with saturated sodium thiosulfate aqueous solution (2 mL), saturated sodium bicarbonate aqueous solution (2 mL), and saturated brine (2 mL), respectively. The mixture was dried over anhydrous sodium sulfate, and the organic phase was concentrated to dryness. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1 to 0:1) to obtain compound 7B. 1 H NMR (400 MHz, CDCl3) δ ppm 8.95 (br d, J=4.02 Hz, 1H), 7.31 (br d, J=9.29 Hz, 1H), 6.76 (br d, J=7.78 Hz, 1H), 5.56 (s, 1H), 5.00-5.10 (m, 1H), 4.71 (d, J=9.03 Hz, 1H), 4.44 (s, 1H), 4.10-4.19 (m, 1H), 4.05(t, J=9.91 Hz, 1H), 3.66 (dd, J=6.40, 10.67 Hz, 1H), 3.28-3.37 (m, 2H), 3.02(q, J=7.11 Hz, 1H), 2.37-2.53 (m, 3H), 1.95-2.25 (m, 5H), 1.81-1.92 (m, 2H), 1.64-1.76 (m, 1H), 1.63-1.78 (m, 4H), 1.39-1.51 (m, 3H), 1.20-1.35 (m, 3H), 1.06-1.08 (m, 1H), 1.08 (s, 9H), 0.56-0.59 (m, 3H), 0.45-0.48 (m, 1H). [M +1] + = 640.2.

[0289] Biological testing

[0290] Experimental Example 1: Evaluation of the in vitro anti-novel coronavirus Mpro protease activity of the test compound

[0291] 1. Experimental materials:

[0292] 1.1 Reagents, consumables and their sources:

[0293] Tris: Sigma;

[0294] EDTA: Sigma;

[0295] NaCl: Sigma;

[0296] 384 well Plate: Perkin Elmer;

[0297] Dimethyl sulfoxide (DMSO): Sigma;

[0298] Substrate(Dabcyl-KTSAVLQSGFRKM-(Edans)):GenScript;

[0299] SARS-CoV-2 Mpro: WuXi AppTec;

[0300] GC376: TargetMol.

[0301] 1.2 Instruments and their sources:

[0302] SpectraMax M2e microplate reader: Molecular Devices;

[0303] Echo 655 Liquid Workstation: Labcyte;

[0304] Benchtop high-speed centrifuge: Eppendorf.

[0305] 2. Experimental Methods:

[0306] 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). The 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 the test concentration of Mpro protein at 25 nM and the test concentration of substrate at 25 μM. The plate was incubated at 30°C for 60 min. The fluorescence signal values ​​at Ex / Em = 340 nm / 490 nm were then detected using a microplate reader. Background wells containing both substrate and compound but without Mpro protein served as controls.

[0307] 3. Data Analysis:

[0308] 1) Calculate the inhibition rate using the following formula:

[0309] Inhibition rate % = [(compound-BG)] 化合物 )-(ZPE-BGZPE )] / [(HPE-BG HPE )-(ZPE-BG ZPE )] * 100%

[0310] # HPE: 100% inhibition control, containing 25 nM Mpro protein + 25 μM substrate + 1 μM GC376

[0311] ZPE: Non-inhibitory control, containing 25 nM Mpro protein + 25 μM substrate, without compounds.

[0312] Compound: Test compound wells. Contains 25 nM Mpro protein + 25 μM substrate + compound.

[0313] BG: Background control wells. Contains 25 μM substrate + compound, Mpro protein-free.

[0314] 2) The inhibition rate data (inhibition rate %) of the compounds were analyzed using GraphPad Prism software using a log(agonist) vs. response-variable slope nonlinear fitting analysis to obtain the IC50 of the compounds. 50 The values ​​and experimental results are shown in Table 1.

[0315] Table 1: In vitro anti-novel coronavirus Mpro protease activity of the test compounds

[0316]

[0317] Conclusion: The compounds of this invention exhibit good in vitro activity against the novel coronavirus Mpro protease.

[0318] Experimental Example 2: Evaluation of the in vitro anticoronavirus activity of compounds using a cytopathic model

[0319] 1. Experimental Materials

[0320] 1.1. Reagents, consumables and their sources

[0321] MEM medium: Sigma;

[0322] L-Glutamine: Gibco;

[0323] Non-essential amino acid: Gibco;

[0324] Double antibody (Penicillin-Streptomycin Solution): HyClone;

[0325] Fetal bovine serum (FBS): ExCell;

[0326] Phosphate-buffered saline (DPBS): Corning;

[0327] 0.25% Pancreatic Enzyme: Gibco

[0328] CellTiter Glo Cell Viability Assay Kit: Promega;

[0329] Remdesivir: MCE;

[0330] 96-well plate: Grenier.

[0331] 1.2. Instruments and Sources

[0332] Microplate reader: BioTek;

[0333] Cell counter: Beckman;

[0334] CO2 incubator: Thermo.

[0335] 1.3. Cells and Viruses

[0336] MRC5 cells and coronavirus HCoV OC43 were purchased from ATCC.

[0337] 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.

[0338] 2. Experimental Methods

[0339] Table 2 Virus testing methods used in this study

[0340]

[0341] 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. 50Cells 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.

[0342] 3. Data Analysis

[0343] The antiviral activity and cytotoxicity of the compounds were expressed as the inhibition rate (%) and cell viability (%) of the compounds against virus-induced cytopathic effects at different concentrations, respectively. The calculation formulas are as follows:

[0344] Inhibition rate (%) = (Test well reading - Average value of virus control) / (Average value of cell control - Average value of virus control) × 100

[0345] Cell viability (%) = (Test well reading - Average value of culture medium control) / (Average value of cell control - Average value of culture medium control) × 100

[0346] The inhibition rate and cell viability of the compound were analyzed using nonlinear fitting analysis with GraphPad Prism. The half-maximal effective concentration (EC50) and half-maximal cytotoxic concentration (CC50) of the compound were calculated. The experimental results are shown in Table 3.

[0347] Table 3: Evaluation of the in vitro anticoronavirus activity of compounds using cytopathic models

[0348]

[0349] Conclusion: The compounds of this invention exhibit good in vitro anti-coronavirus activity at the cellular level and are non-cytotoxic.

[0350] Experimental Example 3: Mouse Pharmacokinetic Study

[0351] This study used male C57BL / 6J mice as test animals and employed LC / MS / MS to quantitatively determine the plasma concentrations of the test compounds administered intravenously and by gavage at different time points in mice to evaluate the pharmacokinetic characteristics of the test drugs in mice.

[0352] The test compound solution was administered orally and by gavage to mice (fasted overnight, 6-8 weeks old). Following intravenous administration, 40 μL of blood was collected from the saphenous vein of mice at 0.083 h, 0.25 h, 0.5 h, 1.0 h, 2.0 h, 4.0 h, 8.0 h, and 24.0 h, and placed in anticoagulant tubes containing EDTA-K2. The plasma was collected by centrifugation at 3200 g for 10 min at 4°C. After processing, the plasma samples were analyzed by LC-MS / MS to determine the drug concentration. Similarly, following gavage administration, 40 μL of blood was collected from the saphenous vein of mice at 0.25 h, 0.5 h, 1.0 h, 2.0 h, 4.0 h, 6.0 h, 8.0 h, and 24.0 h, and placed in anticoagulant tubes containing EDTA-K2. The plasma was collected by centrifugation at 3200 g for 10 min at 4°C. After processing, the plasma samples were analyzed by LC-MS / MS to determine the drug concentration. The experimental results are shown in Tables 4 and 5.

[0353] Table 4. Pharmacokinetic parameters in mice after intravenous injection

[0354]

[0355] Table 5. Pharmacokinetic parameters in mice after gavage administration

[0356]

[0357] ND indicates not detected, NA indicates not detected.

[0358] Conclusion: The compound of the present invention has significantly higher plasma exposure, slower clearance rate, and longer half-life than the reference molecule PF-07321332, and exhibits better pharmacokinetic properties.

Claims

1. A pharmaceutical composition comprising a compound of formula (IV) or a pharmaceutically acceptable salt thereof, and other components. in, R3 is selected from Or R4; Structural unit Selected from , , , , or R2 is selected from C 1-4 Alkyl, C 3-6 cycloalkyl or benzyl; R4 is selected from tert-butyl; Ring B is selected from or .

2. The pharmaceutical composition according to claim 1, wherein, Structural unit Selected from , or .

3. The pharmaceutical composition according to any one of claims 1-2, wherein the compound of formula (IV) has a structure selected from those of formulas (I-1), (IV-1), and (IV-2). 、 、 ， in, R2, R3 and As defined in any one of claims 1-2.

4. The pharmaceutical composition according to claim 3, wherein the compound of formula (IV) has a structure selected from those of formulas (I-1a), (IV-1a), and (IV-2a). 、 、 ， in, R2, R3 and As defined in claim 3.

5. A pharmaceutical composition comprising a compound of the following formula or a pharmaceutically acceptable salt thereof, 。 6. The pharmaceutical composition according to claim 5, wherein the compound is selected from, 。

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