Cycloalkylaminoalkoxy-substituted arylpyrone compounds and uses thereof

Abstract

The present invention discloses novel compounds represented by general formula I and their pharma- ceutically acceptable salts, solvates, and stereoisomers, including mixtures thereof in various ratios, and pharmaceutical compositions containing said compounds, 1a , R 1b , R 1c , R2, R 3a , R 3b , R 4a , R 4b , R5, R6, X1, X2, X3, A ring, etc. have the meanings given in the specification. The invention also relates to a process for the preparation of such compounds, and to the use of said type of compounds and their pharma- ceutically acceptable salts, solvates, stereoisomers, including mixtures in various ratios, particularly in the preparation of antiviral medicaments. [Formula 1] JPEG2025504696000077.jpg4993

Description

[Technical field]

[0001] The present invention is a novel compound and its pharmaceutically acceptable salts, solvates, stereoisomers including mixtures in various ratios and pharmaceutical compositions containing said compound.The present invention further relates to the use of said type of compound and its pharmaceutically acceptable salts, solvates, stereoisomers including mixtures in various ratios, in particular in the preparation of antiviral drugs. [Background technology]

[0002] In this field, a series of antiviral drugs have been developed that target influenza viruses, such as baloxavir marboxil, peramivir, laninamivir octanoate, oseltamivir phosphate, zanamivir, rimantadine, and amantadine. However, there are no drugs that are effective against respiratory viruses such as parainfluenza virus, respiratory syncytial virus, adenovirus, and coronavirus, and it is necessary to use broad-spectrum antiviral drugs such as ribavirin or interferon. At the same time, the drugs commonly used for new coronavirus pneumonia are lopinavir / ritonavir, chloroquine phosphate, arbidol, darunavir, hydroxychloroquine, favipiravir, remdesivir, etc. The above antiviral drugs still have many shortcomings in terms of therapeutic effect and safety, so there is still a need to develop new antiviral therapeutic drugs in this field. Summary of the Invention [Problem to be solved by the invention]

[0003] An object of the present invention is to provide isoflavone derivatives and pharma- ceutically acceptable salts thereof. [Means for solving the problem]

[0004] A first aspect of the present invention provides a compound of formula I, or a pharma- ceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof, [ka] wherein X1 and X2 are each independently selected from the group consisting of O and NR7, wherein R7 is selected from the group consisting of hydrogen, an alkyl group, a substituted alkyl group, an arylalkyl group, a substituted arylalkyl group, a cycloalkyl group, and a substituted cycloalkyl group; X3 is selected from the group consisting of O and NR8, where R8 is selected from the group consisting of hydrogen and an alkyl group; R 1a , R 1b , R 1c are each independently selected from the group consisting of hydrogen, a hydroxyl group, and an alkoxy group; R2 is selected from the group consisting of a substituted or unsubstituted aryl group, or a substituted or unsubstituted heteroaryl group; R 3a , R 3b are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R 4a , R 4b are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R5 is selected from the group consisting of hydrogen or an alkoxy group; R6 is hydrogen, a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C2-C6 acyl group, a substituted or unsubstituted alkoxycarbonyl group, a substituted or unsubstituted carbamoyl group, or (CH2) tR7, where t is 1, 2, 3, 4, 5, or 6, where R7 is selected from the group consisting of a C2-C6 acyl group, a C2-C6 amido group, an aryl group, a heteroaryl group, a cycloalkyl group, a heterocyclyl group, and NR8COR9, where R8 is selected from the group consisting of hydrogen, an alkyl group, and R 9 is selected from the group consisting of alkyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, and heterocyclyl groups; Ring A is a substituted or unsubstituted cycloalkyl group; n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; Here, each substitution means that a hydrogen atom on the group is replaced by one or more substituents selected from the group consisting of cyano groups, halogens, alkyl groups, hydroxyl groups, alkoxy groups, alkenyl groups, alkynyl groups, aryl groups, and heteroaryl groups;

[0005] The compound of formula I is also [ka] Not including, In the above formulas, unless otherwise specified, the alkyl group is a C1-C6 alkyl group and the aryl group is a C6-C 10 The aryl group is a C3-C8 cycloalkyl group, the alkoxy group is a C1-C6 alkoxy group, the alkenyl group is a C2-C6 alkenyl group, the alkynyl group is a C2-C6 alkynyl group, and the heteroaryl group is a 5-12 membered (preferably 5-7 membered) heteroaryl group.

[0006] In another preferred embodiment, X1 is O, and / or X2 is O. In another preferred embodiment, R7 is selected from the group consisting of hydrogen and an alkyl group. In another preferred embodiment, X3 is O. In another preferred embodiment, R 1a , R 1b , R1c are each independently selected from hydrogen. In another preferred example, R2 is a substituted or unsubstituted aryl group, preferably a substituted or unsubstituted phenyl group. In another preferred embodiment, R 3a , R 3b are each independently selected from the group consisting of hydrogen, a C1-C4 alkyl group, and halogen. In another preferred embodiment, R 4a , R 4b are each independently selected from the group consisting of hydrogen, a C1-C4 alkyl group, and halogen. In another preferred example, R5 is selected from hydrogen or a C1-C4 alkoxy group.

[0007] In another preferred embodiment, R6 is hydrogen, a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C2-C6 acyl group, or (CH2) t R7, where t is 1, 2, 3, 4, 5 or 6; R7 is selected from the group consisting of a C2-C6 acyl group, a C2-C6 amido group, an aryl group, and NR8COR9; R8 is selected from the group consisting of hydrogen, an alkyl group; R 9 is selected from the group consisting of an alkyl group and an aryl group. In another preferred example, the A ring is a substituted or unsubstituted C3-C7 cycloalkyl group, preferably a C3-C6 cycloalkyl group. In another preferred embodiment, n is 1, 2, 3 or 4. In another preferred embodiment, R6 is selected from the group consisting of hydrogen and a substituted or unsubstituted alkyl group. In another preferred embodiment, R6 is selected from the group consisting of hydrogen and an unsubstituted alkyl group. In another preferred embodiment, the compound of formula I has the structure of formula II: [ka] Here, R 9a , R 9b , R 9c are each independently hydrogen, a hydroxyl group, a halogen, a cyano group, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C6-C 10 aryl groups, and 5- to 12-membered heteroaryl groups; R 10a , R 10b , R 10c , R 10d , R 10e are each independently selected from the group consisting of hydrogen, a hydroxyl group, a C1-C6 alkoxy group, a halogen, and a cyano group; m is 1, 2, 3, 4 or 5. In another preferred embodiment, the compound according to formula II is selected from the group consisting of: [ka] In another preferred embodiment, the compound of formula I has the structure of formula III: [ka] R 11a , R 11b , R 11c are each independently hydrogen, a hydroxyl group, a halogen, a cyano group, a C1-C6 alkyl group, a C2-C6 alkenyl group, a C2-C6 alkynyl group, a C1-C6 alkoxy group, a C6-C 10 aryl groups, and 5- to 12-membered heteroaryl groups; R 12 is a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C2-C6 acyl group, a substituted or unsubstituted alkoxycarbonyl group, a substituted or unsubstituted carbamoyl group, or (CH2) tR7, where t is 1, 2, 3, 4, 5, or 6, where R7 is selected from the group consisting of a C2-C6 acyl group, a C2-C6 amido group, an aryl group, a heteroaryl group, a cycloalkyl group, a heterocyclyl group, and NR8COR9, where R8 is selected from the group consisting of hydrogen, an alkyl group, and R 9 is selected from the group consisting of alkyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, and heterocyclyl groups; R 13a , R 13b , R 13c , R 13d , R 13e are each independently selected from the group consisting of hydrogen, a hydroxyl group, an alkoxy group, a halogen, and a cyano group.

[0008] In another preferred embodiment, the compound according to formula III is selected from the group consisting of: [ka]

[0009] A second aspect of the present invention provides a pharmaceutical composition, said pharmaceutical composition comprising a compound according to the first aspect, [ka] or a pharma- ceutically acceptable salt, solvate, optically pure isomer or stereoisomer thereof, as well as a pharma- ceutically acceptable carrier.

[0010] A third aspect of the present invention provides the use of a compound of formula I, or a pharma- ceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof, of a pharmaceutical composition according to the second aspect of the present invention, said use being for the preparation of a pharmaceutical composition, [ka] X1 and X2 are each independently selected from the group consisting of O and NR7, where R7 is selected from the group consisting of hydrogen, an alkyl group, a substituted alkyl group, an arylalkyl group, a substituted arylalkyl group, a cycloalkyl group, and a substituted cycloalkyl group; X3 is selected from the group consisting of O and NR8, where R8 is selected from the group consisting of hydrogen and an alkyl group; R 1a , R 1b , R 1c are each independently selected from the group consisting of hydrogen, a hydroxyl group, and an alkoxy group; R2 is selected from the group consisting of a substituted or unsubstituted aryl group, or a substituted or unsubstituted heteroaryl group; R 3a , R 3b are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R 4a , R 4b are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R5 is selected from the group consisting of hydrogen or an alkoxy group;

[0011] R6 is hydrogen, a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C2-C6 acyl group, a substituted or unsubstituted alkoxycarbonyl group, a substituted or unsubstituted carbamoyl group, or (CH2) t R7, where t is 1, 2, 3, 4, 5, or 6, where R7 is selected from the group consisting of a C2-C6 acyl group, a C2-C6 amido group, an aryl group, a heteroaryl group, a cycloalkyl group, a heterocyclyl group, and NR8COR9, where R8 is selected from the group consisting of hydrogen, an alkyl group, and R 9 is selected from the group consisting of alkyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, and heterocyclyl groups; Ring A is a substituted or unsubstituted cycloalkyl group; n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; Here, each substitution means that a hydrogen atom on the group is replaced by one or more substituents selected from the group consisting of cyano groups, halogens, alkyl groups, hydroxyl groups, alkoxy groups, alkenyl groups, alkynyl groups, aryl groups, and heteroaryl groups;

[0012] And the pharmaceutical composition comprises (1) ameliorating, alleviating, or reversing a disease or condition caused by a viral infection; (2) acting as an antagonist of the SARS-Cov-2 virus ACE-2 receptor; (3) acting as a proinflammatory cytokine inhibitor after infection. In another preferred embodiment, the virus is selected from the group consisting of influenza virus, respiratory syncytial virus, coronavirus (eg, SARS-COV-2 virus), and parainfluenza virus. In another preferred embodiment, the infection is a viral infection. In another preferred embodiment, the virus is selected from the group consisting of influenza virus, respiratory syncytial virus, coronavirus (eg, SARS-COV-2 virus), and parainfluenza virus. Effect of the Invention

[0013] It should be understood that within the scope of the present invention, the above technical features of the present invention and the technical features specifically described below (e.g., in the Examples) can be combined with each other to form new or preferred technical solutions, which will not be repeated here due to space limitations. [Brief description of the drawings]

[0014] [Figure 1] A line graph showing the efficacy detection results of compound 4 against the new coronavirus is shown. [Diagram 2]1 is a cell membrane chromatogram of a compound of the present invention. [Diagram 3] This shows the results of calcium imaging of compound 4 in ACE2-293T cells. [Figure 4] 1 shows a bar graph illustrating the effect of compound 4 on INF-β content in lung tissue of a mouse pneumonia model infected with influenza virus FM1. [Diagram 5] 1 shows a bar graph illustrating the effect of Compound 4 on TNF-α content in lung tissue of a mouse pneumonia model infected with influenza virus FM1. [Figure 6] 1 shows a bar graph illustrating the effect of Compound 4 on IL-1 content in lung tissue of a mouse pneumonia model infected with human coronavirus 229E. [Figure 7] 1 shows a bar graph illustrating the effect of Compound 4 on IL-10 content in lung tissue of a mouse pneumonia model infected with human coronavirus 229E. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] After extensive and thorough research, the inventors unexpectedly found that the compound of the present invention has excellent effects in treating diseases caused by various virus infections, and therefore carried out comparative experiments between the compound of the present invention and various known antiviral drugs. Based on this, the present invention was completed. definition Definitions of terms used herein are as follows: Unless otherwise stated, the initial definition of a group or term provided herein applies to that group or term as used herein, either alone or as part of another group.

[0016] The term "substituted" includes any of the following: halogen, nitro, cyano, carboxyl, oxo, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, substituted heterocycle, hydroxyl, alkoxy, aryloxy, alkanoyloxy, aroyloxy, amino, alkanoylamino, arolylamino, arylalkylacylamino, heteroarylalkylacylamino, aminoalkylacylamino, alkylaminoalkylacylamino, dialkylaminoalkylacylamino, alkylamino, aryloxy ... The term "amino group" refers to any substituent mentioned in the specification of the present invention, including, but not limited to, an amino group, an arylalkylamino group, a disubstituted amine (wherein the two amino group substituents are selected from an alkyl group, an aryl group, or an arylalkyl group), an alkanoyl group, a substituted alkanoyl group, an aroyl group, a heteroaroyl group, a carboxyl group, an alkoxycarbonyl group, an aryloxycarbonyl group, an alkylaminocarbonyl group, an arylaminocarbonyl group, an arylalkoxycarbonyl group, an alkylsulfonyl group, an arylsulfonyl group, an arylalkylsulfonyl group, a carbamoyl group, a substituted carbamoyl group, a substituted alkylcarbamoyl group, an amide, a substituted amide, a sulfonamide group, and a substituted sulfonamide group.

[0017] The terms "halogen" or "halo" refer to fluorine, chlorine, bromine and iodine. The term "alkyl group" refers to a straight or branched chain unsubstituted hydrocarbon group having 1 to 20 carbon atoms, preferably 1 to 7 carbon atoms. Examples of "alkyl group" include, but are not limited to, methyl group, ethyl group, 1-propyl group, 2-propyl group, 1-butyl group, 2-butyl group, and the like. The term "substituted alkyl group" refers to an alkyl group substituted with 1 to 4 substituents, including halogen, nitro, cyano, carboxyl, oxo, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocycle, substituted heterocycle, hydroxyl, alkoxy, aryloxy, alkanoyloxy, aroyloxy, amino, alkylamino, arylamino, arylalkylamino, disubstituted amine (wherein the two amino group substituents are selected from alkyl, aryl, or arylalkyl), alkanoyl, substituted alkanoyl, alkoxycarbonyl, arylalkoxycarbonyl, alkylsulfonyl, arylsulfonyl, arylalkylsulfonyl, carbamoyl, substituted carbamoyl, amide, substituted amide, sulfonamide, and substituted sulfonamide.

[0018] The term "alkenyl group" refers to a straight or branched chain hydrocarbon group having 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, and most preferably 2 to 8 carbon atoms and having 1 to 4 double bonds. The term "substituted alkenyl group" refers to an alkenyl group substituted with one or two substituents, such as halogen, nitro, cyano, aryl, substituted aryl, heteroaryl, substituted heteroaryl, alkoxy, aryloxy, alkanoyloxy, aroyloxy, amino, alkylamino, arylamino, arylalkylamino, or disubstituted amine, wherein the two amino group substituents are selected from alkyl, aryl, or arylalkyl groups. The term "alkynyl group" refers to a straight or branched chain hydrocarbon group having 2 to 20 carbon atoms, preferably 2 to 15 carbon atoms, and most preferably 2 to 8 carbon atoms and having 1 to 4 triple bonds. The term "substituted alkynyl group" refers to an alkynyl group substituted by the following substituents: halogen, nitro, cyano, aryl, substituted aryl, heteroaryl, substituted heteroaryl, hydroxyl, alkoxy, aryloxy, alkanoyloxy, aroyloxy, amino, alkylamino, arylamino, arylalkylamino, disubstituted amine, wherein the two amino group substituents are selected from alkyl, aryl, or arylalkyl groups.

[0019] The term "aryl group" refers to a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 12 carbon atoms in the ring portion. Aryl groups include bicyclic groups that contain a ring fused to a saturated or partially unsaturated aromatic ring or an aromatic carbocyclic or heterocyclic ring. Common aryl groups include, but are not limited to, benzene, naphthalene, anthracene, biphenyl groups, 1,2-dihydronaphthalene, 1,2,3,4-tetrahydronaphthyl groups, and the like. The term "substituted aryl group" refers to an aryl group substituted with 1 to 4 substituents, and the substituents can be, for example, a halogen, a nitro group, a cyano group, a ureido group, a carboxyl group, a trifluoromethoxy group, a trifluoromethyl group, an alkyl group, a substituted alkyl group, a cycloalkyl group, a substituted cycloalkyl group, an alkenyl group, a substituted alkenyl group, an alkynyl group, a substituted alkynyl group, an aryl group, a substituted aryl group, a heteroaryl group, a substituted heteroaryl group, a heterocycle, a substituted heterocycle, a hydroxyl group, an alkoxy group, an aryl ... Examples of the aryloxy group include an aryloxy group, an aroyloxy group, an amino group, an alkylamino group, an arylamino group, an arylalkylamino group, a disubstituted amine (wherein the two amino group substituents are selected from an alkyl group, an aryl group, or an arylalkyl group), an alkanoyl group, a substituted alkanoyl group, an alkoxycarbonyl group, an arylalkoxycarbonyl group, an alkylsulfonyl group, an arylsulfonyl group, an arylalkylsulfonyl group, a carbamoyl group, a substituted carbamoyl group, an amide, a substituted amide, a sulfonamide group, and a substituted sulfonamide group. The term "cycloalkyl group" refers to a non-aromatic, saturated or partially unsaturated cyclic hydrocarbon group, which may be substituted with one or more substituents described herein and has a monocyclic ring of 3 to 30 carbon atoms or a bicyclic ring of 7 to 12 carbon atoms. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, cycloheptyl, and cyclooctyl. Exemplary bridged bicyclic cycloalkyl groups include, but are not limited to, bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, and bicyclo[3.2.2]nonane.

[0020] The terms "heterocycle", "heterocyclic" and "heterocyclyl group" refer to an optionally substituted, fully saturated or unsaturated, aromatic or non-aromatic cyclic group, which may be, for example, a 4- to 7-membered monocyclic ring, a 7- to 11-membered bicyclic ring, or a 10- to 15-membered tricyclic ring system having at least one heteroatom in at least one ring containing carbon atoms. Each ring of a heteroatom-containing heterocyclyl group may have 1, 2, or 3 heteroatoms selected from nitrogen, oxygen, and sulfur atoms. The "heterocyclyl group" may be optionally substituted by one or more substituents described herein, and examples of the "heterocyclyl group" include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, morpholino, thiomorpholino, piperazinyl, homopiperazinyl, propylene oxide, imidazolidinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, N-pyridylurea, pyrimidinone, and 1,1-dioxo-thiomorpholinyl. The term "heteroaryl group" refers to a monovalent aromatic group having 5, 6, 7, 8, 9, or 10 ring members, and includes fused systems containing 5 to 20 atoms including one or more heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur, and may be optionally substituted with one or more substituents described herein. Examples of "heteroaryl groups" include, but are not limited to, pyridyl, imidazolyl, imidazopyridyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, thiazolyl, quinolyl, indolyl, and the like.

[0021] The term "oxo" refers to the divalent group =O. The term "carbamoyl group" refers to the group -OC(=O)NH2. The term "amide" refers to a -C(=O)NH2 group. The term "sulfonamide group" refers to a -SO2NH2 group. The terms "substituted carbamoyl group", "substituted amide", and "substituted sulfonamide group" refer to an amide, sulfonamide, or carbamate, respectively, having at least one hydrogen replaced by a group selected from an alkyl group, a substituted alkyl group, an alkenyl group, a substituted alkenyl group, a cycloalkyl group, a substituted cycloalkyl group, an aryl group, a substituted aryl group, a heteroaryl group, a substituted heteroaryl group, a heterocyclyl group, and a substituted heterocyclyl group. The term "acceptable salt" refers to a pharma- ceutically acceptable organic or inorganic salt of a compound of the present invention. Exemplary salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, acid sulfate, isonicotinate, lactate, salicylate, acid citrate, succinate, maleate, fumarate, gluconate, formate, mesylate, and pamoate. An "acceptable salt" can refer to the inclusion of another molecule, such as maleate or other counterion. The counterion stabilizes the charge of the parent compound. An "acceptable salt" can include more than one charged electron, and multiple charged electrons can have multiple counterions.

[0022] When the compound of the invention is a base, the desired "acceptable salt" may be prepared in a suitable manner, for example by treatment of the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or an organic acid such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, salicylic acid, pyranosyl acids such as glucuronic acid or galacturonic acid, α-hydroxy acids such as citric acid or tartaric acid, amino acids such as glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, or sulfonic acids such as methanesulfonic acid or p-toluenesulfonic acid. If the compound of the invention is an acid, the desired "acceptable salt" can be prepared in a suitable manner, for example, by treating the free acid with an inorganic or organic base, such as an amine, an alkali metal hydroxide or an alkaline earth metal hydroxide, etc. Illustrative examples of suitable salts include, but are not limited to, organic salts formed with amino acids, primary, secondary and tertiary amine salts, cyclic amine salts, such as piperidine, morpholine, piperazine, and the like, and inorganic salts formed with sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium. Solvate refers to a complex or complex of one or more solvent molecules and the compound of the present invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethylsulfoxide, ethyl acetate, acetic acid, and ethanolamine. The compound of the present invention can exist in unsolvated form or in solvated form with pharma- ceutically acceptable solvents such as water, ethanol, etc., and therefore the present invention includes both solvated and unsolvated forms.

[0023] The compounds of the present invention contain asymmetric or chiral centers and therefore can exist in different stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and stereoisomers, and mixtures thereof, such as, for example, ceraminic mixtures, form part of the present invention. In the present specification, if the stereochemistry of any particular chiral atom is not determined, all stereoisomers are considered. Furthermore, the present invention relates to all geometric and positional isomers. The compounds of the present invention can exist in different tautomeric forms, and all these forms are included within the scope of the present invention. All stereoisomers of the compounds of the present invention are intended to include in mixture form or in pure or essentially pure form. They can be resolved by physical methods, such as, for example, fractional crystallization, separation or crystallization of diastereomeric derivatives, or separation by chiral column chromatography. The compounds of the present invention can be used alone or in combination with other therapeutic agents. Combination treatments can result in synergistic effects, i.e. the effect achieved when the active ingredients are used together is greater than the sum of the effects obtained when the compounds are used individually. Said combination treatment can be administered simultaneously or in a sequential regimen.When administered sequentially, said combination can be administered in two or more ways.The compounds can be administered together in a single pharmaceutical combination or separately, and when administered separately, they can be administered simultaneously or sequentially in any order. The compounds of the present invention can be administered by any route suitable for the condition to be treated. Suitable routes include, but are not limited to, oral, parenteral (including subcutaneous, intramuscular, intravenous, intraarterial, intradermal), vaginal, intraperitoneal, intrapulmonary and intranasal. It should be understood that the preferred route can vary, for example, depending on the condition of the patient. When the compound is administered orally, it can be formulated into pills, capsules, tablets, etc. with a pharmaceutically acceptable carrier or excipient. When the compound is formulated parenterally, it can be formulated with a pharmaceutically acceptable parenteral carrier.

[0024] The present invention can administer the compound in any convenient formulation form, and the term "formulation" as used herein refers to a dosage form containing the compound of general formula I of the present invention that is advantageous for drug delivery, such as, but not limited to, aqueous solution injection, powder injection, pill, powder, tablet, patch, suppository, emulsion, cream, gel, granule, capsule, aerosol, spray, powder mist, sustained release and controlled release. These pharmaceutical excipients can be those commonly used in various formulations, such as, but not limited to, isotonicity agents, buffer solutions, flavorings, excipients, fillers, binders, disintegrants and lubricants, and can be selected in consideration of compatibility with the above substances, such as emulsifiers, solubilizers, bacteriostatic agents, analgesics and antioxidants, and such excipients can effectively improve the stability and solubility of the compound contained in the composition, or change the release rate and absorption rate of the compound, thereby improving the metabolism of the compound of the present invention in vivo and enhancing the drug administration effect. In addition, they can be used to achieve specific drug administration objectives or methods, such as sustained release drug administration, controlled drug administration, and pulsed drug administration, and the excipients used include, but are not limited to, gelatin, albumin, chitosan, polyether and polyester polymer materials, such as polyethylene glycol, polyurethane, polycarbonate and copolymers thereof, etc. The main symptoms of the so-called "benefits of drug administration" include, but are not limited to, improved therapeutic efficacy, improved bioavailability, reduced toxic side effects, and improved patient compliance.

[0025] In the following examples, only some examples of the present invention are shown to illustrate the method of the present invention. However, these examples do not limit the scope of the present invention, and simple modifications of the manufacturing method of the present invention based on the concept of the present invention are within the protection scope of the present invention. That is, when the present invention is described in combination with the enumerated embodiments, it should be understood that there is no intention to limit the present invention to those embodiments. On the contrary, the present invention is intended to include all changes, modifications and equivalent forms. Those skilled in the art will recognize many methods and materials similar or equivalent to those described in this application that can be used in realizing the present invention. In the examples described below, all temperatures are given in degrees Celsius unless otherwise stated. Reagents are purchased or customized from commercial suppliers such as Sinopharm, Accela, Energy Chemical, TCI, Sigma, etc. unless otherwise stated.

[0026] Example 1. Synthesis of Compound 1 [ka] Weigh out the substrate (0.5mmol) in a 50mL round bottom flask, dissolve in 27mL acetonitrile, add organic amine (5.0mmol), add K2CO3 (1.0mmol), heat to reflux overnight, and monitor the reaction completion by TLC. Spin-dry the solvent, add 50mL water and 100mL ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain the target compound, the yield is 38%.

[0027] Example 2. Synthesis of Compound 2 [ka] Weigh out the substrate (0.5mmol) in a 50mL round bottom flask, dissolve in 27mL acetonitrile, add organic amine (5.0mmol), add K2CO3 (1.0mmol), heat to reflux overnight, and monitor the reaction completion by TLC. Spin-dry the solvent, add 50mL water and 100mL ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain the target compound, the yield is 44%. 1 H NMR(500MHz,CDCl3)δ 8.23(d,J=8.9Hz,1H),7.97(s,1H),7.58(dt,J=3.1,1.8Hz,2H),7.53-7.34(m,3H),6.98(dt,J=28.2,14.1Hz,1H),6.90(d,J=2. 3Hz,1H),4.47-4.06(m,2H),4.03-3.59(m,2H),2.86-2.72(m,1H),2.66-2.47(m,2H),1.18(t,J=7.4Hz,3H),1.04-0.74(m,4H). 13 C NMR(126MHz,DMSO)δ 174.65,163.09,159.02,157.43,153.50,130.10,126.95,124.09,123.36,117.55,115.10,113. 63,101.04,68.31,55.17,47.62,40.02,39.85,39.69,39.52,39.35,39.19,39.02,30.04,6.20.

[0028] Example 3. Synthesis of Compound 3 [ka] Weigh out the substrate (0.5mmol) in a 50mL round bottom flask, dissolve in 27mL acetonitrile, add organic amine (5.0mmol), add K2CO3 (1.0mmol), heat to reflux overnight, and monitor the reaction completion by TLC. Spin-dry the solvent, add 50mL water and 100mL ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain the target compound, the yield is 42%.

[0029] Example 4. Synthesis of Compound 4 [ka] Step 1: Add compound 7-hydroxyisoflavone (0.040 mol), 400 mL of anhydrous acetone, 1,2-dibromoethane (0.403 mol) and potassium carbonate (0.080 mol) to a 1L round bottom flask and heat to reflux overnight. Follow the completion of the reaction by TLC, spin-dry the solvent, add 10 mL of water and 300 mL of ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain 12.98 g of white solid 7-(2-bromoethoxy)isoflavone, the yield is 94%. Step 2: Add compound 7-(2-bromoethoxy)isoflavone (0.5 mmol), 27 mL of CH3CN and cyclopropylamine (1.0 mmol) in turn to a 50 mL round bottom flask, add K2CO3 (1.0 mmol), and heat to reflux overnight. Follow the completion of the reaction by TLC. The solvent was spin-dried, 50mL of water and 100mL of ethyl acetate were added, the organic layer was separated, dried over anhydrous Na2SO4, the organic layer was concentrated to dryness, and subjected to silica gel column chromatography to obtain the target compound, the yield is 53%. 1HNMR(400MHz,DMSO)δ 8.46(s,1H),8.04(d,J=8.8Hz,1H),7.64-7.35(m,5H),7.16(s,1H),7.09(d,J=8.8Hz,1 H),4.17(t,J=5.4Hz,2H),2.99(t,J=5.3Hz,2H),2.23-2.09(m,1H),0.46-0.18(m,4H). 13CNMR(101MHz,DMSO)δ 174.4,163.1,157.4,154.1,132.0,128.9,128.1,127.8,127.0,123.8,117.6,115.1,101.1,68.3,47.6,30.0,6.2. HRMS(ESI):m / z,[M+H] + Calculated value for [M+H] + ):322.1438, Actual value(found):322.1439.

[0030] Example 5. Synthesis of Compound 5 [ka] Add compound 7-(2-bromoethoxy)isoflavone (0.5mmol), 27mL of CH3CN and 1-methyl-cyclopropylamine (1.0mmol) to a 50mL round bottom flask, add K2CO3 (1.0mmol), heat to reflux overnight, and follow the reaction completion by TLC. Spin-dry the solvent, add 50mL of water and 100mL of ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain the target compound, the yield is 37%. 1H NMR(500MHz,CDCl3)δ 8.20(d,J=8.9Hz,1H),7.93(s,1H),7.63-7.49(m,2H),7.47-7.40(m,2H),7.40-7.33(m,1H),6.99(dd,J=8.9,2.4Hz,1H) ,6.85(d,J=2.3Hz,1H),4.14(t,J=5.3Hz,2H),3.13(t,J=5.3Hz,2H),1.32(s,3H),0.71-0.57(m,2H),0.48-0.36(m,2H). 13 C NMR(126MHz,CDCl3)δ 175.73,163.36,157.97,152.80,132.06,129.09,128.58,128.22,127.9 1,125.38,118.61,115.00,100.81,68.75,44.82),34.98,22.10,14.60. HRMS(ESI):C 21 H 22 NO3+ m / z [M+H] + Calculated value: 326.1600, measured value: 336.1529.

[0031] Example 6. Synthesis of Compound 6 [ka] Weigh out the substrate (0.5 mmol) in a 50 mL round bottom flask, dissolve in 27 mL acetonitrile, add organic amine (5.0 mmol), add K2CO3 (1.0 mmol), heat to reflux overnight, and monitor the reaction completion by TLC. Spin-dry the solvent, add 50 mL water and 100 mL ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain the target compound, the yield is 32%.

[0032] Example 7. Synthesis of Compound 7 [ka] Add compound 7-(2-bromoethoxy)isoflavone (0.5mmol), 27mL of CH3CN and cyclohexylamine (1.0mmol) to a 50mL round bottom flask, add K2CO3 (1.0mmol), heat to reflux overnight, and follow the reaction completion by TLC. Spin-dry the solvent, add 50mL of water and 100mL of ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain the target compound, the yield is 45%. 1 H NMR(400MHz,DMSO)δ 8.46(s,1H),8.03(d,J=8.9Hz,1H),7.72-7.30(m,5H),7.17(s,1H),7.09(d,J=8.9Hz,1H),4.16(t,J=5.4Hz,2H ),2.95(t,J=5.4Hz,2H),2.43(t,J=10.0Hz,1H),2.33(d,J=65.9Hz,1H),1.88-1.49(m,5H),1.26-0.98(m,5H). 13 C NMR(101MHz,DMSO)δ 174.4,163.1,157.4,154.2,132.0,128.9,128.1,127.8,127.0,123.8,117.6,115.2,101.1,68.6,56.0,44.7,32.6,25.8,24.4. HRMS(ESI):C 23 H 26 NO3+ m / z [M+H] + Calculated value: 364.1907, Measured value: 364.1909

[0033] Example 8. Synthesis of Compound 8 [ka] Weigh out the substrate (0.5 mmol) in a 50 mL round bottom flask, dissolve in 27 mL acetonitrile, add organic amine (5.0 mmol), add K2CO3 (1.0 mmol), heat to reflux overnight, and monitor the reaction completion by TLC. Spin-dry the solvent, add 50 mL water and 100 mL ethyl acetate, separate the organic layer, dry with anhydrous Na2SO4, concentrate the organic layer to dryness, and subject to silica gel column chromatography to obtain the target compound, the yield is 32%.

[0034] Example 9. Synthesis of Compound 9 [ka] Compound 4 (2 mmol) is added sequentially to a 50 mL round-bottom flask, dissolved in 20 ml of methanol, then paraformaldehyde (20 mmol), sodium cyanoborohydride (10 mmol) and acetic acid (4 mmol) are added, stirred at room temperature overnight, and the reaction is monitored for completion by TLC. The solvent is spin-dried, 100 ml of saturated sodium carbonate solution and 200 mL of ethyl acetate are added, the organic layer is separated, washed with saturated sodium chloride, dried with anhydrous Na2SO4, the organic layer is concentrated to dryness, and subjected to silica gel column chromatography to obtain the target compound, the yield is 72%. 1 H NMR(500MHz,CDCl3)δ 8.22(d,J=5.7Hz,1H),7.86(d,J=91.6Hz,1H),7.43(dd,J=82.0,59.1Hz,5H),7.03(s,1H) ,6.88(s,1H),4.22(s,2H),3.04(s,2H),2.50(s,3H),1.78(s,1H),0.51(d,J=16.3Hz,4H). 13 C NMR(126MHz,CDCl3)δ 175.63,163.31,157.91,152.67,132.02,129.01,128.49,128.13,127. 77,125.26,118.46,115.03,100.76,66.55,56.67,43.70,38.66,6.91. HRMS(ESI):C 21 H22 NO3+ m / z [M+H] + Calculated value: 336.16, measured value: 336.1672.

[0035] Example 10. Synthesis of Compound 10 [ka] Compound 4 (2 mmol) is added sequentially to a 50 mL round-bottom flask, dissolved in 20 ml pyridine, then acetic anhydride (20 mmol) is added, stirred at room temperature overnight, and the reaction is monitored by TLC for completion. The solvent is spin-dried, 100 ml 1M hydrochloric acid and 200 mL ethyl acetate are added, the organic layer is separated, washed with 1M hydrochloric acid, washed with saturated sodium bicarbonate, washed with saturated sodium chloride, dried with anhydrous Na2SO4, the organic layer is concentrated to dryness, and subjected to silica gel column chromatography to obtain the target compound, the yield is 81%. 1 H NMR(500MHz,CDCl3)δ 8.25(t,J=29.2Hz,1H),7.96(s,1H),7.55(t,J=18.2Hz,2H),7.45(t,J=7.4Hz,2H),7.39(t,J=7.3Hz,1H),6.98(dt,J=21.9,10.9Hz, 1H),6.90(d,J=1.9Hz,1H),4.47-4.11(m,2H),3.78(dt,J=30.5,5.4Hz,2H),2.99-2.63(m,1H),2.35-1.98(m,3H),1.09-0.68(m,4H). 13 C NMR(126MHz,CDCl3)δ 175.63,174.16,163.08,157.95,152.76,131.98,129.02,128.51,128.16, 127.87,125.29,118.59,114.91,100.72,66.40,46.77,32.17,22.82,9.79.

[0036] Example 11. Synthesis of Compound 11 [ka] Compound 4 (2 mmol) is added sequentially to a 50 mL round-bottom flask, dissolved in 20 ml pyridine, then propionic anhydride (20 mmol) is added, stirred at room temperature overnight, and monitored for reaction completion by TLC. The solvent is spin-dried, 100 ml 1M hydrochloric acid and 200 mL ethyl acetate are added, the organic layer is separated, washed with 1M hydrochloric acid, washed with saturated sodium bicarbonate, washed with saturated sodium chloride, dried with anhydrous Na2SO4, the organic layer is concentrated to dryness, and subjected to silica gel column chromatography to obtain the target compound, the yield is 83%. 1 H NMR(500MHz,CDCl3)δ 8.23(d,J=8.9Hz,1H),7.97(s,1H),7.58(dt,J=3.1,1.8Hz,2H),7.53-7.34(m,3H),6.98(dt,J=28.2,14.1Hz,1H),6.90(d,J=2. 3Hz,1H),4.47-4.06(m,2H),4.03-3.59(m,2H),2.86-2.72(m,1H),2.66-2.47(m,2H),1.18(t,J=7.4Hz,3H),1.04-0.74(m,4H). 13 C NMR(126MHz,CDCl3)δ 177.35,175.70,163.17,158.02,152.79,132.03,129.07,128.56,128.21,127 .93,125.36,118.62,114.97,100.75,66.53,47.02,31.41,27.56,9.73,9.23.

[0037] Example 12. Synthesis of Compound 12 [ka] Compound 4 (2 mmol) is added sequentially to a 50 mL round-bottom flask, dissolved in 20 ml of methanol, then acetone (20 mmol), sodium cyanoborohydride (10 mmol) and acetic acid (4 mmol) are added, stirred at room temperature overnight, and monitored for reaction completion by TLC. The solvent is spin-dried, 100 ml of saturated sodium carbonate solution and 200 mL of ethyl acetate are added, the organic layer is separated, washed with saturated sodium chloride, dried with anhydrous Na2SO4, the organic layer is concentrated to dryness, and subjected to silica gel column chromatography to obtain the target compound, the yield is 75%. 1 H NMR(500MHz,CDCl3)δ 8.21(d,J=8.9Hz,1H),7.95(s,1H),7.60-7.52(m,2H),7.44(dd,J=10.2,4.7Hz,2H),7.41-7.35(m,1H),6.99(dd,J=8.9,2 .3Hz,1H),6.86(d,J=1.6Hz,1H),4.15(s,2H),3.14(s,1H),3.03(s,2H),1.98(s,1H),1.10(s,6H),0.49(d,J=37.0Hz,4H). 1 H NMR(500MHz,MeOD)δ 8.25(s,1H),8.13(d,J=9.3Hz,1H),7.58-7.52(m,2H),7.44(t,J=7.4Hz,2H),7.41-7.36(m,1H),7.11-7.06(m,2H),4.25(t,J=6.1 Hz,2H),3.22-3.13(m,1H),3.07(t,J=6.1Hz,2H),2.11-1.92(m,1H),1.14(d,J=6.7Hz,6H),0.59-0.52(m,2H),0.49-0.43(m,2H). 13 C NMR(126MHz,CDCl3)δ 175.71,163.51,158.03,152.69,132.10,129.08,128.55,128.17,127.84, 125.33,118.44,115.09,100.76,68.10,53.25,49.13,34.54,18.97,7.23. HRMS(ESI):C 23 H 26NO3+ m / z [M+H] + Calculated value: 364.1913, measured value: 364.1906.

[0038] The chemical structures shown in the tables or figures below are as follows: [ka]

[0039] Example 13: Therapeutic effect of the compound of the present invention on a mouse pneumonia model infected with influenza A H1N1 virus FM1 strain 1. Test Materials 1.1. Test drugs: Compound 4, Compound F1, Compound F2, Compound F3 1.2. Positive control Ribavirin Granules: Production unit: Sichuan Baili Pharmaceutical Co., Ltd. Indications: This product is suitable for viral pneumonia, bronchitis, and cutaneous herpes virus infections caused by RS virus. Main ingredient: Ribavirin. Properties: White or off-white soluble granules. Specification: 50 mg / bag, 18 bags / box. Usage and dosage: Oral, 150 mg, 3 times a day. Storage conditions: Store in a sealed, dry place. [Table A-1] [Table A-2] [Table A-3]

[0040] 2. Test Method 2.1. Dose design and preparation [Table A-4]

[0041] 2.2. Modeling and Administration ICR mice were collected and randomly divided into normal control group, model control group and ribavirin control group according to body weight level. Compound 4, compound F1, compound F2 and compound F3 were each divided into two dose groups, each group had 10 mice. Except for the normal control group, the mice were lightly anesthetized with ether and 15 LD 50 Infect the animals intranasally with 30 μl of H1N11 influenza virus liquid FM1 strain per animal. Administration begins on the day of infection, and the animals are gavaged once a day for 4 consecutive days according to 0.2 ml / 10 g each time. The normal control group and the model control group are gavaged with distilled water under similar conditions. On the 5th day, the animals are weighed and then dissected, the lungs are weighed, and the lung index and the inhibition rate of the lung index are calculated. The results are statistically processed using t-tests for comparison between groups. Lung index (%) = Lung wet weight (g) x 100 / body weight (g)

number

[0042] 3. Test Results [Table 1] According to the results in Table 1, after the mice were infected with the H1N1 influenza virus FM1 strain virus, the lung index of the mice increased obviously, with a significant difference (P<0.01) compared with the normal control group; after treatment with compound 4, compound F1, compound F2, and compound F3 for 4 days starting from the day of infection, compound 4 can obviously reduce the lung index of the mice after infection in the 50mg / kg / d dose group, with a significant difference (P<0.05) compared with the model control group, indicating that 50mg / kg of compound F1 tends to reduce the lung index, and compound F2 and compound F3 have no obvious effect on the mouse lung index. The efficacy of compound 4 is stronger than that of compound F1, which indicates that the 7-position substituent has a very important effect on the anti-influenza virus efficacy of such isoflavone derivatives; the efficacy of compound 4 is stronger than that of compound F2 and compound F3, which indicates that the position and structure of the substituent have a very important effect on the anti-influenza virus efficacy of such isoflavone derivatives.

[0043] Example 14: Therapeutic effect of the compound of the present invention on a mouse pneumonia model infected with influenza A H1N1 virus FM1 strain, PR8 1. Test Materials 1.1. Test drugs: Compound 4, Compound F4, Compound F5, Compound 5, Compound 7 1.2. Positive control Oseltamivir phosphate capsules (Tamiflu), production unit: Hoffmann-Roche Co., Ltd., Basel, Switzerland. Indications: Used for the treatment of influenza A and B in adults and children aged 1 year or older, and for the prevention of influenza A and B in adults and adolescents aged 13 years or older. Main ingredient: Oseltamivir phosphate. Properties: This product is a gray and light yellow capsule, and the contents are a white to yellowish white powder that may contain lumps. Specifications: Oseltamivir, 75 mg / tablet, calculated as 10 tablets / box. Dosage and administration: Adults and adolescents aged 13 years or older, 75 mg each time, administered orally twice a day for a total of 5 days. Storage conditions: Store in a sealed container. [Table B-1] [Table B-2] [Table B-3]

[0044] 2. Test Method 2.1. Dose design and preparation [Table B-4]

[0045] 2.2 Test procedure ICR mice were collected and randomly divided into normal control group, model control group and oseltamivir control group according to body weight level. Compound 4, Compound F4, Compound F5, Compound 5 and Compound 7 were divided into two dose groups, each group had 10 mice. Except for the normal control group, the mice were lightly anesthetized with ether and 15 LD 50 The animals are infected intranasally with 30 μl of H1N11 influenza virus fluid (FM1 / PR8 strain) per animal. Administration begins on the day of infection, and the animals are gavaged once a day for 4 consecutive days at 0.2 ml / 10 g each time. The normal control group and the model control group are gavaged with distilled water under similar conditions. On the 5th day, the animals are weighed and then dissected, the lungs are weighed, and the lung index and the inhibition rate of the lung index are calculated. The results are statistically processed using t-tests for comparison between groups. Lung index (%) = Lung wet weight (g) x 100 / body weight (g)

number

[0046] 3. Test Results [Table 2] According to the results in Table 2, after mice are infected with H1N1 influenza virus FM1 strain virus, the lung index of mice increases obviously, with a significant difference (P<0.01) compared with the normal control group; after treatment with the compounds of the present invention for 4 days starting from the day of infection, compound 4 can obviously reduce the lung index of mice after infection at a dose of 40mg / kg / d, with a significant difference (P<0.01) compared with the model control group, and the inhibition rate of the lung index is 54.04%, indicating that the 40mg / kg / d dose of compound 7 tends to reduce the lung index. However, compound F4, compound F5, and compound 5 have no effect on reducing the lung index of mice after infection. The efficacy of compounds F4, F5, 5 and 7 was reduced compared to that of compound 4, where compounds F4 and F5 are metabolites of compound 4 in vivo, compound 5 is a derivative having a 2-(1-methylcyclopropylamino)ethoxy group substituent at the 7-position, and compound 7 is a derivative having a 2-(cyclohexylamino)ethoxy group substituent at the 7-position, proving that the 2-(cyclopropylamino)ethoxy group substituent at the 7-position has a very significant effect on the efficacy of such isoflavone derivatives. [Table 3] According to the results in Table 3, after the mice were infected with the H1N1 influenza virus PR8 strain, the lung index of the mice increased obviously, with a significant difference (P<0.01) compared with the normal control group; after treatment with compound 4 for 4 days starting from the day of infection, compound 4 can obviously reduce the lung index of the mice after infection at a dose of 15 mg / kg / d, with a significant difference (P<0.01) compared with the model control group, indicating that the inhibition rate of the lung index is 60.27%, and the inhibition rate of oseltamivir at a dose of 27.5 mg / kg / d is 58.89%.

[0047] Example 15: Therapeutic effect of the compound of the present invention on a mouse pneumonia model infected with human coronavirus 229E 1. Test Materials 1.1. Test drugs: Compound 4, Compound F1, Compound F2, Compound F3. 1.2. Positive drug: Chloroquine phosphate tablets, Sichuan Sunnyhope Pharmaceutical Co., Ltd. Specification: 0.25g / tablet, Dosage and administration: 0.5g / 60kg / d, orally administered. 1.3. Laboratory animals [Table C-1] 1.4. Virus strains and cells: Human coronavirus 229E (HCoV-229E) was provided by the Institute of Medical Biotechnology, Chinese Academy of Medical Sciences, and passaged in our laboratory and stored in a refrigerator at -80℃. Human fetal lung fibroblast MRC5 was purchased from Beijing Bena Culture Collection, Institute of Biological Technology, and passaged in our laboratory and stored in liquid nitrogen. 1.5. Test Equipment [Table C-2] 2. Test Method 2.1. Drug Dose Design and Preparation [Table C-3]

[0048] 2.2 Virus passaging 25 cm grown on monolayer MRC-5 cells 2 Take the culture flask, discard the culture medium, and use cell maintenance liquid to wash the cell surface three times. Then add 5ml of cell maintenance liquid, then add 200μl of HCoV-229E virus liquid, and culture in a 37℃, 5% CO2 incubator for 72-96 hours. Observe the cell lesion status under an inverted microscope every day until 80% of the cells show obvious lesions (CPE). Place the cell culture flask in a -80℃ low-temperature refrigerator for freezing and storage. The virus liquid is frozen and thawed three times before being used for virus titer measurement. 2.3. Measurement of viral titer Take the 96-well plate with monolayer MRC5 cells grown, discard the culture medium, and wash the cells three times using cell maintenance liquid. Then inoculate HCoV-229E virus liquid with different titers at 10-fold dilution rates (10-1 to 10-8), a total of eight dilutions, at 100 μl / well, and create four duplicate wells for each dilution, while setting up normal cell controls. Place the 96-well plate in a 37°C, 5% CO2 incubator and culture for 72 to 96 hours. Observe the cell lesion status under an inverted microscope every day and record the cell lesion status of each well. 50% cell lesion concentration (TCID 50 ) to calculate 2.4. Construction and administration of a human coronavirus pneumonia mouse model BALB / c mice were randomly divided into a normal control group, a model control group, and a chloroquine phosphate control group according to body weight. Compound 4, Compound F1, Compound F2, and Compound F3 were divided into two dose groups, with 10 mice in each group, half male and half female. Except for the normal control group, the mice in each group were lightly anesthetized with ether and then injected with 100 TCID 50 Intranasally infect the mice using HCoV-229E, 50 μl / mouse, once every other day, for a total of two infections. On the day of the first infection, each treatment group is administered once a day for four consecutive days. After weighing the body weight one day after the final administration, the lungs are dissected and weighed, and the lung index and inhibition rate of the mice are calculated. Lung index = wet lung weight (g) x 100 / body weight (g)

number

[0049] 3. Test Results [Table 4] According to the results in Table 4, after the mice were infected with human coronavirus 229E, the mouse lung index of the model control group increased significantly, with a significant difference (P<0.01) compared with the normal control group, and compound 4 significantly reduced the mouse lung index in the 50 mg / kg and 40 mg / kg dose groups, with a significant difference (P<0.05) compared with the model control group, with the inhibition rates being 41.00% and 41.85%, respectively. Compounds F1, F2, and F3 have no obvious effect on the mouse lung index. The efficacy of compound 4 is stronger than that of compound F1, which indicates that the 7-position substituent has a very important effect on the anti-coronavirus efficacy of such isoflavone derivatives, and the efficacy of compound 4 is stronger than that of compounds F2 and F3, which indicates that the position and structure of the substituent have a very important effect on the anti-coronavirus efficacy of such isoflavone derivatives.

[0050] Example 16: Effect of compounds of the present invention on a murine pneumonia model infected with coronavirus 229E The test reagents, test equipment, and test method are the same as those described above. Test samples: Compound 4, Compound F4, Compound F5, Compound 5, Compound 7 Animal certificate: BALB / c mouse 1100112011066562881 / 718 [Table 5] According to the results in Table 5, after the mice were infected with human coronavirus 229E, the mouse lung index increased obviously, with a significant difference (P<0.01) compared with the normal control group; after the compounds of the present invention were administered for 4 days starting from the day of infection, compound 4 at a dose of 30 mg / kg / d, 15 mg / kg / d, and 7.5 mg / kg / d can obviously reduce the lung index of the mice after infection, with all significant differences (P<0.01, P<0.05) compared with the model control group, and the inhibition rates of the lung index were 86.46%, 76.52%, and 44.34%, respectively; compound 7 at a dose of 15 mg / kg / d can reduce the lung index, with a significant difference (P<0.05) compared with the model control group, and the inhibition rate was 41.56%, indicating that compound F4, compound F5, and compound 5 have no effect on the reduction of the lung index of the mice after infection. The efficacy of compounds F4, F5, 5, and 7 was reduced compared to that of compound 4, demonstrating that the 2-(cyclopropylamino)ethoxy group substituent at the 7-position has a very significant effect on the anti-coronavirus efficacy of such isoflavone derivatives.

[0051] Example 17: Therapeutic effect of Compound 4 on a mouse pneumonia model infected with human coronavirus OC43 The test reagents, test equipment, and test method are the same as those described above. Test sample: Compound 4 Animal certificate: BALB / c mouse 110011211110038632 [Table 6] According to the results in Table 6, after the mice were infected with human coronavirus OC43 virus, the mouse lung index increased obviously, with a significant difference (P<0.01) compared with the normal control group; after treatment with compound 4 for 4 days starting from the day of infection, compound 4 can obviously reduce the lung index of the mice after infection at a dose of 15 mg / kg / d, with all significant differences (P<0.05) compared with the model control group, showing that the inhibition rate of the lung index is 47.43%, and the inhibition rate of chloroquine phosphate at a dose of 90 mg / kg / d is 42.72%.

[0052] Example 18: Effect of Compound 4 of the Present Invention on a Mouse Pneumonia Model Infected with Parainfluenza Virus 1. Test materials 1.1. Test drug: Compound 4 1.2. Ribavirin Granules, Production Unit: Sichuan Baili Pharmaceutical Co., Ltd. Indications: This product is suitable for viral pneumonia, bronchitis, and cutaneous herpes virus infections caused by respiratory syncytial virus. Main ingredient: Ribavirin. Properties: White or off-white soluble granules. Specification: 50 mg / bag, 18 bags / box. Dosage and administration: Oral, 150 mg, 3 times a day. Storage conditions: Store in a sealed, dry place. 1.3. Test animals [Table D-1] 1.4. Viral strains: Parainfluenza viruses were purchased from ATCC, USA, and stored at -80°C in our laboratory. 1.5. Test Equipment [Table D-2]

[0053] 2. Test Method 2.1. Dose design and preparation [Table D-3] 2.2. Modeling and Administration Sixty ICR mice, SPF level, weighing 13-15g, were randomly divided into six groups according to body weight: normal control group, model control group, ribavirin group, and three dose groups of Compound 4. Each group contained 10 mice, half male and half female. Except for the normal control group, the mice in each group were lightly anesthetized with ether and then infused with 100TCID 50Each mouse was intranasally infected with 45 μl of Sendai strain parainfluenza virus. After infection, the mice in each group were administered the corresponding drugs, and the normal control group and model control group were administered distilled water under the same conditions for 4 consecutive days. On the 5th day, the mice were weighed, and the lung tissue was isolated and the lungs were weighed. Pulmonary index = lung weight × 100 / body weight,

number

[0054] Example 19: Compound 4 of the present invention has anti-new coronavirus (SARS-CoV-2) effect in vitro 1. Test Materials 1.1. Test drug: Compound 4 1.2. Cell line: VeroE6 cells are stored in the Virus Laboratory of the National Laboratory of Respiratory Diseases, Guangzhou Respiratory Health Research Institute. 1.3.Virus strain: SARS-CoV-2, titer is TCID 50 = 10-6 / 100μL and stored at -80℃ in the National Key Laboratory for Biosafety Detection (P3 Laboratory) of the Satellite Quarantine Institute of Guangzhou Customs Technical Center. The virus titer used is 100TCID 50 It is. 2. Test Method 2.1. Sample Preparation Drug names, experimental concentrations and groupings [Table E]

[0055] 2.2. Antiviral experiments of test drugs [1] In a sterile 96-well culture plate, add 100 µL of 2 x 10 5 VeroE6 cells were added at a concentration of 1000 cells / mL and cultured at 37°C and 5% CO2 for 24 hours. [2] 100TCID 50 Add 100 μL of virus solution per well and allow to adsorb for 2 hours in a 37℃, 5% CO2 incubator. [3] After 2 hours, discard the cell culture medium in the 96-well culture plate, dilute the test drug to each concentration in Table 1, prepare three duplicate wells for each concentration, and add 100 μl of the above drug solution to each well. [4] At the same time, set up a cell control, a blank control (solvent control), and a virus control (negative control). [5] Incubate the cells in a 37°C, 5% CO2 incubator for 3-4 days. [6] Cellular lesions (CPE) were observed under an optical microscope, and the degree of cellular lesions was recorded according to the following 6-level scale: "-" indicates the absence of lesions, "±" indicates less than 10% cellular lesions, "+" indicates approximately 25% cellular lesions, "++" indicates approximately 50% cellular lesions, "+++" indicates approximately 75% cellular lesions, and "++++" indicates 75% or more cellular lesions. The median effective concentration (IC 50 ) is calculated. Criteria for determining efficacy: The concentration that inhibits viral CPE by 50% is defined as the effective concentration. 3. Test Results By observing cytopathic cell injury (CPE) and recording the experimental results, the median effective concentration (IC) was calculated using the Reed-Muench method or GraphPad Prism 5.0. 50 ) was calculated and the experimental results are as follows: [Table 8] The efficacy detection results of compound 4 against the new coronavirus are shown in Figure 1. The results in Table 8 show that drug compound 4 has an inhibitory effect on the cytopathology of novel coronavirus (SARS-CoV-2) infection in Vero E6 cells in vitro.

[0056] Example 20: Study on the in vitro mechanism of action of compound 4 of the present invention against SARS-Cov-2 1. Binding effect to ACE2 receptor of 293T cells Test objective: To observe the binding effect of the test sample with the ACE-2 receptor and find out the possible mechanism of action. Equipment and Materials Test drugs: Compound 4, Compound F1, Compound F2, Compound F3 SHIMADZU LC-2040C 3D high-performance liquid chromatograph (Japan), Sartorius 1712 type electronic analytical balance, Fram FCR1002-UF ultrapure water preparation machine (Qingdao Fram Science and Technology Co., Ltd.), SHZ-D (III) type circulating water vacuum pump (Guinyi City Yingyu Yuhua Equipment Factory), SB-5200DTD ultrasonic cleaning device (Ningbo Scientz Bioscience Technology Co., Ltd.), pipette (Germany Eppendorf). A 293T cell line that highly expresses the ACE2 receptor is constructed in the laboratory. Methanol was chromatography grade, water was ultrapure, and all other reagents were analytical grade. Test Method Binding effect of samples with 293T cell ACE2 receptor Sample preparation Accurately weigh out 5 mg of each of compound 4, compound F1, compound F2, and compound F3 of the present invention, dissolve them completely in chromatography grade methanol, transfer them to a 10 mL measuring flask, and adjust the volume to obtain a pharmaceutical stock solution of the compound series of the present invention with a concentration of 0.5 mg / mL. Store these in a dark place in a refrigerator to reserve them, and dilute them to the appropriate concentration when used. Prepare the stock solution fresh every week. CMC Chromatography Conditions ACE2-293T / CMC cell membrane chromatography column (10×2.0 mm), mobile phase is water, flow rate is 0.2 mL / min, column temperature is 37° C., sample size is 10 μL, DAD detection. Positive control: chloroquine phosphate. Test results The cell membrane chromatogram of the compound of the present invention is as shown in FIG. [Table 9] 2. Evaluation of the functional effects of compound 4 on ACE2-293 T cells Test method: Intracellular Ca 2+ Detection of changes in flow rate: Insert the cells into a 96-well plate, discard the medium after overnight, wash with CIB, add Fluo-3, incubate the AM incubation solution in an incubator for 40 minutes, then discard the incubation solution, wash with CIB, and under a fluorescent microscope, add drugs to each well sequentially after discarding the CIB, observe the changes in the fluorescence intensity of the cells before and after adding the drugs, observe continuously for 2 minutes under a fluorescent microscope, and add the drugs to stimulate 10 seconds after the photographing observation. Test results: Intracellular Ca 2+ Changes in flux: Compound 4 can stimulate calcium mobilization in ACE2-293T cells at 10 μg / mL, and as the administration concentration increases, the degree of calcium fluctuation in ACE2-293T cells is dose-dependently enhanced. The results of calcium imaging of Compound 4 in ACE2-293T cells are shown in Figure 3.

[0057] Example 21: Effect of Compound 4 on the content of inflammatory factors in lung tissue of mice infected with influenza virus Lung tissue samples: Therapeutic effect of compound 4 on a mouse pneumonia model infected with influenza virus H1N1 / FM1 strain Test method: Detection of inflammatory factors in mouse lung tissue (Elisa method) Tissue homogenate sample: After weighing the mouse lung tissue, collect the mouse lung tissue and store it at -4℃. After weighing 50mg of lung tissue and adding 500μL of physiological saline, homogenize the tissue using an ultrasonic cell disrupter and centrifuge at -4℃ and 1000xg for 10 minutes using a low-temperature high-speed centrifuge. Aspirate the supernatant and then dispense it into aliquots, and store it in a refrigerator at -80℃ for storage. Avoid repeated freezing and thawing. Sample preparation: The sample is detected after diluting it two-fold with diluent: 50 μL serum + 50 μL diluent. Remove the microwell plate from the sealed bag equilibrated to room temperature and add 100 μL per well of each standard, experimental sample or quality control product at various concentrations to the corresponding wells. Seal the reaction wells with sealing tape and incubate at room temperature for 2 hours. Wash the plate with washing solution and repeat 4 times. After the last plate wash is completed, invert the plate, dab dry all remaining liquid on absorbent paper and add 100 μL of enzyme-labeled detection antibody to each microwell. Seal the reaction wells with sealing tape and incubate at room temperature for 2 hours. Repeat the plate washing procedure in step 4, add 100 μL of color-developing substrate to each microwell and incubate at room temperature for 30 minutes. Be careful to avoid light. Measure the absorbance value at 450 nm using a microplate reader within 30 minutes after adding 100 μL of stop solution to each microwell. Calculate the results. The results in Figures 4 and 5 show that after mice were infected with influenza virus H1N1 / FM1 strain, the IFN-β and TNF-a contents in the mouse lung tissue increased, with significant difference (p<0.01) compared with the normal control group, and after 4 days of administration, compound 4 can obviously reduce the IFN-β and TNF-a contents in the mouse lung tissue after infection, with significant difference (p<0.01) compared with the model control group.

[0058] Example 22: Effect of Compound 4 on the content of inflammatory factors in lung tissue of mice infected with coronavirus 229E Lung tissue samples: Therapeutic effects on a mouse pneumonia model infected with human coronavirus 229E According to the results in Figures 6 and 7, after mice were infected with human coronavirus 229E, the IL-1 and IL-10 contents in the mouse lung tissue increased, with a significant difference (p<0.01) compared with the normal control group; after 4 days of administration, compound 4 could obviously reduce the IL-1 and IL-10 contents in the mouse lung tissue after infection, with a significant difference (p<0.01) compared with the model control group.

[0059] Example 23. Therapeutic effect of the compound of the present invention on a mouse pneumonia model infected with human coronavirus 229E 1. Test materials 1.1. Test drugs: Compound 4, Compound 9, Compound 10, Compound 11, Compound F1. 1.2. Positive drug: Chloroquine phosphate tablets, Sichuan Sunnyhope Pharmaceutical Co., Ltd. Specification: 0.25g / tablet, Dosage and administration: 0.5g / 60kg / d, orally administered. 1.3. Laboratory animals [Table F-1] 1.4. Virus strains and cells: Human coronavirus 229E (HCoV-229E) was provided by the Institute of Medical Biotechnology, Chinese Academy of Medical Sciences, and passaged in our laboratory and stored in a refrigerator at -80℃. Human fetal lung fibroblast MRC5 was purchased from Beijing Bena Culture Collection, Institute of Biological Technology, and passaged in our laboratory and stored in liquid nitrogen. 1.5. Test Equipment [Table F-2]

[0060] 2. Test Method 2.1. Drug Dose Design and Preparation [Table F-3] 2.2. Virus Passage 25 cm grown on monolayer MRC-5 cells 2Take the culture flask, discard the culture medium, and use cell maintenance liquid to wash the cell surface three times. Then add 5ml of cell maintenance liquid, then add 200μl of HCoV-229E virus liquid, and culture in a 37℃, 5% CO2 incubator for 72-96 hours. Observe the cell lesion status under an inverted microscope every day until 80% of the cells show obvious lesions (CPE). Place the cell culture flask in a -80℃ low-temperature refrigerator for freezing and storage. The virus liquid is frozen and thawed three times before being used for virus titer measurement. 2.3. Measurement of viral titer The 96-well plate containing monolayer MRC5 cells was taken, the culture medium was discarded, and the cells were washed three times using cell maintenance solution. Then, HCoV-229E virus liquids with different titers were diluted 10-fold (10 -1 ~10 -8 ), a total of 8 dilutions were inoculated at 100 μl / well, and 4 duplicate wells were prepared for each dilution, and normal cell controls were also set up. The 96-well plate was placed in a 37°C, 5% CO2 incubator and cultured for 72-96 hours. The cell lesions were observed every day under an inverted microscope, and the cell lesions in each well were recorded. The 50% cell lesion concentration (TCID 50 ) to calculate 2.4. Construction and administration of a human coronavirus pneumonia mouse model BALB / c mice were randomly divided into a normal control group, a model control group, and a chloroquine phosphate control group according to body weight. Compound 4, Compound 9, Compound 10, and Compound 11 were each a single dose group, with 10 mice in each group, half male and half female. Except for the normal control group, the mice in each group were lightly anesthetized with ether and then 100 TCID 50 Intranasally infect the mice using HCoV-229E, 50 μl / mouse, once every other day, for a total of two infections. On the day of the first infection, each treatment group is administered once a day for four consecutive days. After weighing the body weight one day after the final administration, the lungs are dissected and weighed, and the lung index and inhibition rate of the mice are calculated. Lung index = wet lung weight (g) x 100 / body weight (g)

number

[0061] 3. Test Results [Table 10] The results in Table 10 show that after the mice were infected with human coronavirus 229E, the mouse lung index of the model control group increased significantly, with a significant difference (P<0.05) compared with the normal control group, and the mouse lung index of the 7.5 mg / kg / d dose group of compound 4 and compound 9 decreased significantly, with a significant difference (P<0.05) compared with the model control group.

[0062] Example 24. Effects of the compounds of the present invention on a mouse pneumonia model infected with parainfluenza virus 1. Test Materials 1.1. Test drugs: Compound 4, Compound 9, Compound 10, Compound 11, Compound 12, Compound 2. 1.2. Ribavirin Granules, Production Unit: Sichuan Baili Pharmaceutical Co., Ltd. Indications: This product is suitable for viral pneumonia, bronchitis, and cutaneous herpes virus infections caused by respiratory syncytial virus. Main ingredient: Ribavirin. Properties: White or off-white soluble granules. Specification: 50 mg / bag, 18 bags / box. Dosage and administration: Oral, 150 mg, 3 times a day. Storage conditions: Store in a sealed, dry place. 1.3. Test animals [Table G-1] 1.4. Viral strains: Parainfluenza viruses were purchased from ATCC, USA, and stored at -80°C in our laboratory. 1.5. Test Equipment [Table G-2] 1.6. Test location: ABSL-2 Laboratory of the Institute of Traditional Chinese Medicine, China Academy of Traditional Chinese Medicine

[0063] 2. Test Method 2.1. Dose design and preparation [Table G-3] 2.2. Modeling and Administration 150 ICR mice, SPF level, weighing 13-15g, were randomly divided into 15 groups according to their body weight: normal control group, model control group, ribavirin group, and two dose groups of Compound 4, Compound 9, Compound 10, Compound 11, Compound 12, and Compound 2. Each group contained 10 mice, half male and half female. Except for the normal control group, the mice in each group were lightly anesthetized with ether and then infused with 100TCID 50 Each mouse was intranasally infected with 45 μl of Sendai strain parainfluenza virus. After infection, the mice in each group were administered the corresponding drugs, and the normal control group and model control group were administered distilled water under the same conditions for 4 consecutive days. On the 5th day, the mice were weighed, and the lung tissue was isolated and the lungs were weighed. Pulmonary index = lung weight in grams × 100 / body weight in grams

number

[0064] Example 25. Therapeutic effect of the compound of the present invention on mouse pneumonia models infected with influenza A viruses H1N1 / PR8 and H1N1 / FM1 1. Test materials 1.1. Test drugs: Compound 9, Compound 12, Compound 2. 1.2. Positive control Oseltamivir phosphate capsules (Tamiflu), production unit: Hoffmann-Roche Co., Ltd., Basel, Switzerland. Indications: Used for the treatment of influenza A and B in adults and children aged 1 year or older, and for the prevention of influenza A and B in adults and adolescents aged 13 years or older. Main ingredient: Oseltamivir phosphate. Properties: This product is a gray and light yellow capsule, and the contents are a white to yellowish white powder that may contain lumps. Specifications: Oseltamivir, 75 mg / tablet, calculated as 10 tablets / box. Dosage and administration: Adults and adolescents aged 13 years or older, 75 mg each time, administered orally twice a day for a total of 5 days. Storage conditions: Store in a sealed container. 1.3. Test animals [Table H-1] 1.4.Virus strains [Table H-2] 1.5. Test Equipment [Table H-3] 1.6. Test location: ABSL-2 Laboratory of the Institute of Traditional Chinese Medicine, China Academy of Traditional Chinese Medicine

[0065] 2. Test Method 2.1. Dose design and preparation [Table H-4] 2.2. Test Procedure ICR mice were collected and randomly divided into nine groups according to body weight level, including a normal control group, a model control group, an oseltamivir phosphate control group, and two dose groups of the drugs Compound 9, Compound 12, and Compound 2, each with 10 mice. Except for the normal control group, the mice were lightly anesthetized with ether and 15 LD 50The mice are intranasally infected with 30 μl of H1N11 / PR8 influenza virus liquid per mouse. The administration starts on the day of infection, and the mice are gavaged once a day for 4 consecutive days according to 0.2 ml / 10 g each time. The normal control group and the model control group are gavaged with distilled water under the same conditions. On the 5th day, the mice are weighed and then dissected, and the lungs are weighed, and the lung index and the inhibition rate of the lung index are calculated. The results are statistically processed using t-test for comparison between groups. Lung index (%) = Lung wet weight (g) x 100 / body weight (g)

number

[0066] 3. Test Results [Table 12] The results in Table 12 show that after the mice were infected with influenza A H1N1 / PR8 virus, the mouse lung index increased obviously, with a significant difference (P<0.01) compared with the normal control group; after the mice were treated with compound 9, compound 12, and compound 2 for 4 days starting from the day of infection, the lung index decreased significantly, with a significant difference (P<0.01) compared with the model control group. [Table 13] The results in Table 13 show that after the mice were infected with influenza A H1N1 / FM1 virus, the mouse lung index increased obviously, with a significant difference (P<0.01) compared with the normal control group; after the mice were treated with compound 9, compound 12, and compound 2 for 4 days starting from the day of infection, the lung index decreased significantly, with a significant difference (P<0.01) compared with the model control group.

[0067] Example 26: Therapeutic effect of compound 12 on a mouse pneumonia model infected with human coronavirus 229E The experimental materials and methods were the same as those in Example 23. [Table 14] The results in Table 14 show that after the mice were infected with human coronavirus 229E, the mouse lung index of the model control group increased significantly, with a significant difference (P<0.05) compared with the normal control group, and the mouse lung index of the 15 mg / kg / d and 7.5 mg / kg / d dose groups of compound 12 decreased significantly, with a significant difference (P<0.01, P<0.05) compared with the model control group.

[0068] Example 27: Acute toxicity test of compound 4 in mice 1. Test Method Seventy ICR mice, weighing 17-21g, were randomly divided into seven groups according to their fasting weights: a normal control group, and six dose groups of the test drug (compound 4) at 475, 633, 844, 1125, 1500, and 2000mg / kg, with 10 mice in each group, half male and half female. After fasting for about 15 hours, each treatment group was given a single oral gavage dose of 40ml / kg, and the control group was given an equal amount of 0.5% sodium carboxymethylcellulose solution once a day, and the animals were observed for death for 14 consecutive days after administration. Immediately after administration, the animals are observed for the presence or absence of toxic reactions, the symptoms of toxicity and their severity, the time of appearance and disappearance of toxicity, side effects are recorded, and gross dissection of dead animals is performed, including the observation of the animals' fur, eyes, ears, mouth, nose, genital orifices, anus and its mucous membranes. 2. Experimental Results 2.1. Observation of mouse mortality and side effects Compound 4 was administered orally to animals once daily at doses of 475, 633, 844, 1125, 1500, and 2000 mg / kg of the test article, and the main toxic reaction observed was acute death. In the groups dosed with 1125 mg / kg or more of the test product, no animal deaths occurred after administration, and in the groups dosed with 844 mg / kg or less of the test product, all animal deaths occurred within 4 days after administration, with the peak of deaths occurring within 24 to 48 hours after administration. The mortality rates of animals in the six dose groups were 0%, 0%, 0%, 20%, 80% and 100%, respectively. 2.2. In addition to acute death, other macroscopic side effects mainly include decreased spontaneous activity, immobility, ataxia, tremors, convulsions, proneness, loss of orthostatic reflex and piloerection, etc., and the 633, 844, 1125, 1500 and 2000 mg / kg dose groups of the test product showed various degrees of changes in the above indications, with incidence rates of 40%, 60%, 80%, 100% and 100%, respectively, occurring within 60 minutes to 5 days after administration. In the 475 mg / kg dose group of the test product, there were no macroscopic side effects. Then, until the end of the 14-day observation period, no abnormalities were found in the animals' general condition, activity, walking, breathing, eating, drinking, defecation, fur, etc. In summary, the LD50 calculated using the Bliss method is 1299.054 mg / kg of the test article, with 95% confidence limits of 1152.937-1463.820 mg / kg of the test article, the maximum non-lethal dose is 844 mg / kg of the test article, and the maximum non-adverse effect dose is 475 mg / kg of the test article.

[0069] Example 28: Acute toxicity study of compound 12 in mice 1. Test Method Thirty ICR mice, weighing 17-21g, were randomly divided into three groups according to their fasting weight: a normal control group and test drug (compound 12) groups at doses of 2000mg and 4000mg / kg, with 10 mice in each group, half male and half female. After fasting for about 15 hours, each treatment group was forced to receive a single dose of 40ml / kg of carboxymethylcellulose sodium solution, and the control group was forced to receive an equal amount of 0.5% carboxymethylcellulose sodium solution once a day, and the animals were observed for death for 14 consecutive days after administration. Immediately after administration, the animals are observed for the presence or absence of toxic reactions, the symptoms of toxicity and their severity, the time of appearance and disappearance of toxicity, side effects are recorded, and gross dissection of dead animals is performed, including the observation of the animals' fur, eyes, ears, mouth, nose, genital orifices, anus and its mucous membranes. 2. Experimental Results Compound 12 was administered orally to the animals once a day at doses of 2000 and 4000 mg / kg, and the animals were observed from immediately after administration until the end of the 14-day observation period, and no abnormalities were found in the animals' general condition, activity, walking, breathing, eating, drinking, defecation, fur, etc. No animals died during the experiment.

[0070] In conjunction with the above examples, the therapeutic effect of the compound of the present invention on the mouse pneumonia model infected with influenza virus H1N1 / FM1 strain, as well as the effect of the compound of the present invention on the mouse pneumonia model infected with human coronavirus 229E, the efficacy of compound 4 is stronger than that of the less potent modified precursor 7-hydroxyisoflavone, indicating that the modification strategy of the 7-hydroxyisoflavone at the 7-position in the present invention has a very important effect on the antiviral efficacy of such isoflavone derivatives, and the efficacy of compound 4 is stronger than that of compound F2 (glycosylation strategy derivative at the 7-position), compound F4 (alkoxyalkoxy group derivative at the 7-position), and compound F5 (amide alkoxy group derivative at the 7-position), indicating that the antiviral effect of compound 4 obtained by the conversion strategy of the cycloalkylaminoalkoxy group strategy is superior. The antiviral efficacy of compound 4 is stronger than that of compound 5 and compound 7, indicating that the 2-(cyclopropylamino)ethoxy group substituted derivative at the 7-position is more effective. This indicates that the position and structure of the substituent have a very important effect on the antiviral efficacy of 7-hydroxyisoflavone derivatives, and experiments also show that the antiviral efficacy of introducing a 2-(cyclopropylamino)ethoxy group substituent at the 7-position is the best. Compounds 9 and 12 derived from compound 4 also have very strong pharmacological effects, but they are weaker than those of compounds 10 and 11 derived from compound 4, which indicates that the structure of the cyclopropyl group and R6 in general formula (I) has a very important effect on the antiviral efficacy. When retaining a cyclopropylamine group substituent, derivatives in which R6 is an alkyl group or a substituted alkyl group have very strong pharmacological activity. At the same time, the results of preliminary toxicity studies showed that the LD50 of compound 4 in acute toxicity experiments in mice was 1299.054 mg, and the maximum concentration of compound 12 that can be administered orally by force was 4000 mg / kg, and no animal deaths occurred, indicating that compared with compound 4, compound 12 has a larger safety margin and better pharmacological efficacy, which indicates that the derivatives in which R6 is an alkyl group or a substituted alkyl group not only maintain better pharmacological activity, but also have higher safety. All documents mentioned in the present invention are incorporated by reference in this application as if each document was incorporated by reference individually. Furthermore, after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and these equivalents are also included in the scope defined by the appended claims of this application.

Claims

1. A compound of formula I, or a pharma- ceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof, 【Chemistry 1】 X １ , X ２ are each independently O, NR ７ wherein R ７ is selected from the group consisting of hydrogen, an alkyl group, a substituted alkyl group, an arylalkyl group, a substituted arylalkyl group, a cycloalkyl group, and a substituted cycloalkyl group; X ３ , O, NR ８ wherein R ８ is selected from the group consisting of hydrogen, alkyl groups, R １ａ , R １ｂ , R １ｃ are each independently selected from the group consisting of hydrogen, a hydroxyl group, and an alkoxy group; R ２ is selected from the group consisting of a substituted or unsubstituted aryl group, or a substituted or unsubstituted heteroaryl group; R ３ａ , R ３ｂ are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R ４ａ , R ４ｂ are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R ５ is selected from the group consisting of hydrogen or an alkoxy group; R ６ is a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C ２ -C ６ an acyl group, a substituted or unsubstituted alkoxycarbonyl group, a substituted or unsubstituted carbamoyl group, or (CH ２ ) ｔ R ７ where t is 1, 2, 3, 4, 5 or 6, and ７ is C ２ -C ６ Acyl group, C ２ -C ６ Amide group, aryl group, heteroaryl group, cycloalkyl group, heterocyclyl group, NR ８ C.O.R. ９ wherein R ８ is selected from the group consisting of hydrogen, alkyl groups, R ９ is selected from the group consisting of alkyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, and heterocyclyl groups; Ring A is a substituted or unsubstituted cycloalkyl group; n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; Here, each substitution means that a hydrogen atom on the group is replaced by one or more substituents selected from the group consisting of cyano groups, halogens, alkyl groups, hydroxyl groups, alkoxy groups, alkenyl groups, alkynyl groups, aryl groups, and heteroaryl groups; The compound of formula I is also 【Chemistry 2】 Not including, In the above formulas, unless otherwise stated, the alkyl group is C １ -C ６ The alkyl group is C ６ -C １０ The aryl group is a cycloalkyl group. ３ -C ８ The cycloalkyl group is an alkoxy group. １ -C ６ The alkoxy group is an alkenyl group. ２ -C ６ The alkynyl group is C ２ -C ６ The compound represented by formula I, characterized in that the aryl group is an alkynyl group and the heteroaryl group is a 5-12 membered (preferably 5-7 membered) heteroaryl group, or a pharma- ceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof.

2. X １ is O, and / or X ２ is O, In another preferred embodiment, the R ７ is selected from the group consisting of hydrogen, alkyl groups, In another preferred embodiment, X ３ is O, In another preferred embodiment, R １ａ , R １ｂ , R １ｃ are each independently selected from hydrogen. The compound of claim 1.

3. R ２ is a substituted or unsubstituted aryl group, preferably a substituted or unsubstituted phenyl group; In another preferred embodiment, R ３ａ , R ３ｂ are each independently hydrogen, C １ -C ４ selected from the group consisting of alkyl groups, halogens; In another preferred embodiment, R ４ａ , R ４ｂ are each independently hydrogen, C １ -C ４ selected from the group consisting of alkyl groups, halogens; In another preferred embodiment, R ５ is hydrogen or C １ -C ４ alkoxy groups; In another preferred embodiment, R ６ is a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C ２ -C ６ Acyl group, or (CH ２ ) ｔ R ７ where t is 1, 2, 3, 4, 5, or 6; R ７ is C ２ -C ６ Acyl group, C ２ -C ６ Amide group, aryl group, NR ８ C.O.R. ９ wherein R ８ is selected from the group consisting of hydrogen, alkyl groups, R ９ is selected from the group consisting of an alkyl group and an aryl group; In another preferred embodiment, the A ring is a substituted or unsubstituted C ３ -C ７ is a cycloalkyl group, preferably C ３ -C ６ is a cycloalkyl group, In another preferred embodiment, n is 1, 2, 3, or 4; In another preferred embodiment, the R ６ is selected from the group consisting of hydrogen, substituted or unsubstituted alkyl groups; In another preferred embodiment, the R ６ is selected from the group consisting of hydrogen and unsubstituted alkyl groups. The compound of claim 1.

4. The compound of formula I has the structure shown in formula II: 【Chemistry 3】 R ９ａ , R ９ｂ , R ９ｃ are each independently a hydrogen atom, a hydroxyl group, a halogen atom, a cyano group, １ -C ６ Alkyl group, C ２ -C ６ Alkenyl group, C ２ -C ６ Alkynyl group, C ６ -C １０ aryl groups, and 5- to 12-membered heteroaryl groups; R １０ａ , R １０ｂ , R １０ｃ , R １０ｄ , R １０ｅ are each independently hydrogen, a hydroxyl group, or C １ -C ６ is selected from the group consisting of an alkoxy group, a halogen, and a cyano group; m is 1, 2, 3, 4 or 5; The compound of claim 1.

5. The compound according to formula II is characterized in that it is selected from the group consisting of: The compound according to claim 4. 【Chemistry 4】

6. The compound of formula I has the structure shown in formula III: 【Chemistry 5】 R １１ａ , R １１ｂ , R １１ｃ are each independently a hydrogen atom, a hydroxyl group, a halogen atom, a cyano group, １ -C ６ Alkyl group, C ２ -C ６ Alkenyl group, C ２ -C ６ Alkynyl group, C １ -C ６ Alkoxy group, C ６ -C １０ aryl groups, and 5- to 12-membered heteroaryl groups; R １２ is a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C ２ -C ６ an acyl group, a substituted or unsubstituted alkoxycarbonyl group, a substituted or unsubstituted carbamoyl group, or (CH ２ ) ｔ R ７ where t is 1, 2, 3, 4, 5 or 6, and ７ is C ２ -C ６ Acyl group, C ２ -C ６ Amide group, aryl group, heteroaryl group, cycloalkyl group, heterocyclyl group, NR ８ C.O.R. ９ wherein R ８ is selected from the group consisting of hydrogen, alkyl groups, R ９ is selected from the group consisting of alkyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, and heterocyclyl groups; R １３ａ , R １３ｂ , R １３ｃ , R １３ｄ , R １３ｅ are each independently selected from the group consisting of hydrogen, a hydroxyl group, an alkoxy group, a halogen, and a cyano group. The compound of claim 1.

7. The compound according to formula III is characterized in that it is selected from the group consisting of: The compound according to claim 6. 【Chemistry 6】

8. 1. A pharmaceutical composition comprising: The pharmaceutical composition comprises a compound according to any one of claims 1 to 7, 【Chemistry 7】 or a pharma- ceutically acceptable salt, solvate, optically pure isomer or stereoisomer thereof, and a pharma- ceutically acceptable carrier.

9. 9. Use of a compound of formula I, or a pharma- ceutically acceptable salt, solvate, optically pure isomer, stereoisomer or mixture thereof, or a pharmaceutical composition according to claim 8, comprising: Used in the preparation of a pharmaceutical composition, 【Chemistry 8】 X １ , X ２ are each independently O, NR ７ wherein R ７ is selected from the group consisting of hydrogen, an alkyl group, a substituted alkyl group, an arylalkyl group, a substituted arylalkyl group, a cycloalkyl group, and a substituted cycloalkyl group; X ３ , O, NR ８ wherein R ８ is selected from the group consisting of hydrogen, alkyl groups, R １ａ , R １ｂ , R １ｃ are each independently selected from the group consisting of hydrogen, a hydroxyl group, and an alkoxy group; R ２ is selected from the group consisting of a substituted or unsubstituted aryl group, or a substituted or unsubstituted heteroaryl group; R ３ａ , R ３ｂ are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R ４ａ , R ４ｂ are each independently selected from the group consisting of hydrogen, an alkyl group, and a halogen; R ５ is selected from the group consisting of hydrogen or an alkoxy group; R ６ is a hydrogen atom, a substituted or unsubstituted alkyl group, a substituted or unsubstituted arylalkyl group, an aminoalkyl group, a substituted or unsubstituted cycloalkyl group, a substituted or unsubstituted heterocyclyl group, a substituted or unsubstituted C ２ -C ６ an acyl group, a substituted or unsubstituted alkoxycarbonyl group, a substituted or unsubstituted carbamoyl group, or (CH ２ ) ｔ R ７ where t is 1, 2, 3, 4, 5 or 6, and ７ is C ２ -C ６ Acyl group, C ２ -C ６ Amide group, aryl group, heteroaryl group, cycloalkyl group, heterocyclyl group, NR ８ C.O.R. ９ wherein R ８ is selected from the group consisting of hydrogen, alkyl groups, R ９ is selected from the group consisting of alkyl groups, aryl groups, heteroaryl groups, cycloalkyl groups, and heterocyclyl groups; Ring A is a substituted or unsubstituted cycloalkyl group; n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; Here, each substitution means that a hydrogen atom on the group is replaced by one or more substituents selected from the group consisting of cyano groups, halogens, alkyl groups, hydroxyl groups, alkoxy groups, alkenyl groups, alkynyl groups, aryl groups, and heteroaryl groups; And the pharmaceutical composition comprises (1) ameliorating, alleviating or reversing a disease or condition caused by a viral infection; (2) acting as a SARS-Cov-2 virus ACE-2 receptor antagonist; (3) The above use, characterized in that it is used for an application selected from the group consisting of acting as an inhibitor of inflammatory cytokines after infection.

10. the virus is selected from the group consisting of influenza virus, respiratory syncytial virus, coronavirus (e.g., SARS-COV-2 virus), and parainfluenza virus; In another preferred embodiment, the infection is a viral infection, In another preferred embodiment, the virus is selected from the group consisting of influenza virus, respiratory syncytial virus, coronavirus (e.g., SARS-COV-2 virus), and parainfluenza virus.

10. The use according to claim 9.

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