Macrocyclic compound and application thereof in medicine

Abstract

The invention belongs to the technical field of medicines, and relates to a macrocyclic compound and application thereof in medicines. Specifically, the invention relates to a compound shown in a general formula (I) or a stereoisomer, a tautomer, a nitrogen oxide, a solvate, a metabolite, a pharmaceutically acceptable salt or a prodrug thereof, and a pharmaceutical composition containing the compound, the invention relates to a compound and a pharmaceutical composition thereof, and application of the compound and the pharmaceutical composition in preparation of a medicine for preventing and / or treating diseases or symptoms caused by coronavirus, especially diseases or symptoms caused by novel coronavirus (SARS-CoV-2), and the variables in the formula (I) are defined in the specification. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and relates to the use of a macrocyclic compound in preparing a drug for treating and / or preventing a disease or symptom caused by a coronavirus, and in particular, to the use of a macrocyclic compound in preparing a drug for treating and / or preventing a disease or symptom caused by a novel coronavirus (SARS-CoV-2). The present invention also relates to the use of a pharmaceutical composition composed of these macrocyclic compounds and other antiviral drugs in preparing a drug for treating and / or preventing a disease or symptom caused by a coronavirus such as SARS-CoV-2. Background Art

[0002] Coronaviruses (CoVs) belong to the order Nidovirales, family Coronaviridae, subfamily Orthocoronavirus; this subfamily includes four genera: α, β, γ, and δ. Before 2019, only six CoVs could infect humans and cause respiratory diseases: HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1 only induced mild upper respiratory tract diseases, and in rare cases, caused severe infections in infants, young people, and the elderly; SARS-CoV and MERS-CoV can infect the lower respiratory tract and cause severe respiratory syndrome in humans.

[0003] The new coronavirus is the seventh coronavirus discovered so far that can infect humans and cause disease. It belongs to the beta coronavirus, which is highly contagious and can also infect the lower respiratory tract and cause pneumonia. The WHO named it severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The genome of SARS-CoV-2 is a single-stranded positive-strand RNA with a genome size of 29903nt, containing a 5′-end cap structure and a 3′-end polyadenylic acid tail (A new coronavirus associated with human respiratory disease in China [J]. Nature, 2020, 579 (7798): 265-269). Under an electron microscope, the virus is mostly spherical, some are polymorphic, and have a diameter of 60 to 140nm. The virus particles have obvious spikes, about 9 to 12nm, which makes the virus appear corona-shaped.

[0004] The clinical manifestations of novel coronavirus pneumonia are mainly fever, fatigue, and dry cough. A small number of patients have symptoms such as nasal congestion, runny nose, myalgia, and diarrhea. Severe cases often develop dyspnea after one week, and severe cases rapidly progress to acute respiratory distress syndrome, septic shock, difficult-to-correct metabolic poisoning, coagulation dysfunction, and multiple organ failure.

[0005] 3Clpro (3C-like cysteine ​​protease, 3CL protease) has become a potential effective target for the development of anti-broad-spectrum coronavirus drugs. Currently reported 3Clpro inhibitors (3CL protease inhibitors) include covalent peptidomimetic inhibitors and non-covalent small molecule inhibitors. Although peptidomimetic covalent inhibitors have significant inhibitory activity against 3Clpro, covalent inhibitors have poor target selectivity, unpredictable toxic side effects, and poor metabolic stability. Non-covalent small molecule inhibitors are a better choice, but the non-covalent small molecule inhibitors reported so far are very scarce, and have problems such as single structure, weak enzyme inhibition activity, and poor drugability. Therefore, there is still an urgent need to develop new, safe and effective drugs for the treatment and / or prevention of SARS-CoV-2. Summary of the invention

[0006] The macrocyclic compound described in the present invention is a class of 3CL protease inhibitors, which have good inhibitory activity against coronaviruses, especially SARS-CoV-2, and have the advantages of good pharmacokinetic properties, good solubility, good liver microsomal stability, basically no induction effect on liver drug enzymes and low toxicity. It has good application prospects in drugs for treating and / or preventing diseases or symptoms caused by coronaviruses, especially SARS-CoV-2.

[0007] In one aspect, the present invention relates to a compound, which is a compound as represented by formula (I) or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof of the compound as represented by formula (I),

[0008]

[0009] Where L is C 2-8 Alkylene or C 2-12 Alkenylene, wherein the C 2-12 Alkylene and C 2-8 Each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 groups independently selected from deuterium, F, Cl, Br and C 1-4 substituted by an alkyl substituent;

[0010] Each R 1 , R 2 , R 3 , R 4 H, deuterium, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Haloalkyl;

[0011] R 5 H, deuterium, F, Cl, Br, -CN, C 1-6Alkyl, deuterated C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Haloalkyl or C 3-6 Cycloalkyl;

[0012] R 6 C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Haloalkyl;

[0013] R is H, deuterium, F, Cl, Br, -CN, -NO2, -COOH, -C 1-6 Alkylene -COOH, amino, -C(=O)OC 1-6 Alkyl, -C 1-6 Alkylene-C(=O)OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 6-14 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6 Cycloalkyl, heterocyclic group consisting of 3-10 ring atoms, bicyclic heteroaryl consisting of 9-10 ring atoms, -C 2-6 Alkenylene-C(=O)OR 6 or -C(=O)CH2-OC(=O)R 7 , wherein the -C 1-6 Alkylene -COOH, amino, -C(=O)OC 1-6 Alkyl, -C 1-6 Alkylene-C(=O)OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 6-14 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6 Cycloalkyl, heterocyclic group consisting of 3-10 ring atoms, bicyclic heterocyclic group consisting of 9-10 ring atoms, -C 2-6 Alkenylene-C(=O)OR 6 C 2-6 Alkenylene and -C(=O)CH2-OC(=O)R 7 The CH2 in each of the above is independently unsubstituted or substituted by 1, 2 or 3 independently selected from deuterium, F, Cl, Br, -OH, CN, C 1-6 Alkyl, C 1-6Haloalkyl, -COOH, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy and C 1-6 substituted with a hydroxyalkoxy substituent;

[0014] R 7 C 6-14 aryl, a monocyclic heteroaryl consisting of 5-6 ring atoms, or a bicyclic heteroaryl consisting of 9-10 ring atoms, wherein the C 6-14 The aryl group, the monocyclic heteroaryl group consisting of 5-6 ring atoms and the bicyclic heteroaryl group consisting of 9-10 ring atoms are each independently unsubstituted or substituted with 1, 2, 3 or 4 R w1 replaced by;

[0015] Each R w1 are independently deuterium, F, Cl, Br, -CN, -NO2, -COOH, amino, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 Halogenated alkoxy.

[0016] In some embodiments of the present invention, wherein L is C 4-12 Alkylene or C 4-12 Alkenylene, wherein the C 4-12 Alkylene and C 4-12 Each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 groups independently selected from deuterium, F, Cl, Br and C 1-3 The alkyl group is substituted with an alkyl substituent.

[0017] In some embodiments of the present invention, L is C 6-12 Alkylene or C 6-12 Alkenylene, wherein the C 6-12 Alkylene and C 6-12 Each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl.

[0018] In some embodiments of the present invention, L is C 8-11 Alkylene or C 8-11 Alkenylene, wherein the C 8-11 Alkylene and C 8-11 Each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl.

[0019] In some embodiments of the present invention, L is C 9-10 Alkylene or C 9-10 Alkenylene, wherein the C9-10 Alkylene and C 9-10 Each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl.

[0020] In some embodiments of the present invention, L is -(CH2)9-, -(CH2) 10 -, -(CH2)6-CH=CHCH2-, -(CH2)7-CH=CHCH2-, -(CH2)5-CH=CHCH2CH2- or -(CH2)-CH2CH=CHCH2-, wherein -(CH2)9-, -(CH2) 10 -, -(CH2)6-CH=CHCH2-, -(CH2)7-CH=CHCH2-, -(CH2)5-CH=CHCH2CH2- and -(CH2)-CH2CH=CHCH2- are each independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl.

[0021] In some embodiments of the present invention, each R 1 , R 2 , R 3 and R 4 H, deuterium, C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 2-4 Alkenyl or C 1-4 Halogenated alkyl.

[0022] In some embodiments of the present invention, each R 1 , R 2 , R 3 and R 4 and are independently H, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CD3, CHD2, -CH2CD3, vinyl, propenyl, allyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2 or -CH2CH2CF3.

[0023] In some embodiments of the present invention, R 5 H, deuterium, F, Cl, Br, -CN, C 1-4 Alkyl, deuterated C 1-4 Alkyl, C2-4 Alkenyl, C 1-4 haloalkyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0024] In some embodiments of the present invention, R 5 It is H, F, Cl, Br, -CN, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CD3, CHD2, -CH2CD3, vinyl, propenyl, allyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0025] In some embodiments of the present invention, R 6 C 1-4 Alkyl, C 2-4 Alkenyl or C 1-4 Halogenated alkyl.

[0026] In some embodiments of the present invention, R 6 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, vinyl, propenyl, allyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2 or -CH2CH2CF3.

[0027] In some embodiments of the present invention, R is H, deuterium, F, Cl, Br, -CN, -NO2, -COOH, -C 1-4 Alkylene -COOH, amino, -C(=O)OC 1-4 Alkyl, -C 1-4 Alkylene-C(=O)OC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 6-12 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6Cycloalkyl, heterocyclic group consisting of 3-6 ring atoms, bicyclic heteroaryl consisting of 9-10 ring atoms, -C 2-4 Alkenylene-C(=O)OR 6 or -C(=O)CH2-OC(=O)R 7 , wherein the -C 1-4 Alkylene -COOH, amino, -C(=O)OC 1-4 Alkyl, -C 1-4 Alkylene-C(=O)OC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, C 1-4 Hydroxyalkyl, C 6-12 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6 Cycloalkyl, heterocyclic group consisting of 3-6 ring atoms, bicyclic heteroaryl consisting of 9-10 ring atoms, -C 2-4 Alkenylene-C(=O)OR 6 C 2-4 Alkenylene and -C(=O)CH2-OC(=O)R 7 The CH2 in each of the above is independently unsubstituted or substituted by 1, 2 or 3 independently selected from deuterium, F, Cl, Br, -OH, CN, C 1-4 Alkyl, C 1-4 Haloalkyl, -COOH, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy and C 1-4 The hydroxyl group is substituted with an alkoxy group.

[0028] In some embodiments of the present invention, R is H, deuterium, F, Cl, Br, -CN, -NO2, -COOH, -C 1-2 Alkylene -COOH, amino, -C(=O)OC 1-3 Alkyl, -C 1-2 Alkylene-C(=O)OC 1-3Alkyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-l-propoxy, 2-butoxy, C 1-3 Haloalkoxy, C 1-3 hydroxyalkyl, phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, propyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, thietanyl, oxirane, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, dihydropyridine, morpholinyl, thiomorpholinyl, piperazinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothiophenyl, indolyl, purinyl, quinolinyl, isoquinolinyl, -CH=CH-C(=O)OR 6 、-CH=CHCH2-C(=O)OR 6 、-CH2CH=CH-C(=O)OR 6 、-CH2CH=CHCH2-C(=O)OR 6 、-CH=CHCH2CH2-C(=O)OR 6 、-CH2CH2CH=CH-C(=O)OR 6 or -C(=O)CH2-OC(=O)R 7 , wherein the -C 1-2 Alkylene -COOH, amino, -C(=O)OC 1-3 Alkyl, -C 1-2 Alkylene-C(=O)OC 1-3Alkyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, C 1-3 Haloalkoxy, C 1-3 hydroxyalkyl, phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, propyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, thietanyl, oxirane, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, dihydropyridine, morpholinyl, thiomorpholinyl, piperazinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothiophenyl, indolyl, purinyl, quinolinyl and isoquinolinyl, -CH=CH-C(=O)OR 6 CH=CH、-CH=CHCH2-C(=O)OR 6 CH=CHCH2、-CH2CH=CH-C(=O)OR 6 CH2CH=CH、-CH2CH=CHCH2-C(=O)OR 6 CH2CH=CHCH2、-CH=CHCH2CH2-C(=O)OR 6 CH=CHCH2CH2、-CH2CH2CH=CH-C(=O)OR 6 CH2CH2CH=CH and -C(=O)CH2-OC(=O)R 7wherein CH2 is independently unsubstituted or replaced by 1, 2 or 3 independently selected from deuterium, F, Cl, Br, -OH, CN, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -COOH, C 1-3 Hydroxyalkyl, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy and C 1-3 The hydroxyl group is substituted with an alkoxy group.

[0029] In some embodiments of the present invention, R 7 is phenyl, naphthyl, a monocyclic heteroaryl consisting of 5 ring atoms, a monocyclic heteroaryl consisting of 6 ring atoms, a bicyclic heteroaryl consisting of 9 ring atoms, or a bicyclic heteroaryl consisting of 10 ring atoms, wherein the phenyl, naphthyl, monocyclic heteroaryl consisting of 5 ring atoms, monocyclic heteroaryl consisting of 6 ring atoms, bicyclic heteroaryl consisting of 9 ring atoms, and bicyclic heteroaryl consisting of 10 ring atoms are each independently unsubstituted or substituted with 1, 2, 3 or 4 R w1 replaced.

[0030] In some embodiments of the present invention, R 7 is phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothienyl, indolyl, purinyl, quinolyl or isoquinolyl, wherein the phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridinyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothienyl, indolyl, purinyl, quinolyl and isoquinolyl are each independently unsubstituted or substituted with 1, 2, 3 or 4 R w1 replaced.

[0031] In some embodiments of the present invention, each R w1are independently deuterium, F, Cl, Br, -CN, -NO2, -COOH, amino, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy or C 1-4 Halogenated alkoxy.

[0032] In some embodiments of the present invention, each R w1 are independently deuterium, F, Cl, Br, -CN, -NO2, -COOH, amino, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy or C 1-3 Halogenated alkoxy.

[0033] In some embodiments of the present invention, the compound of the present invention is a compound as shown in formula (II) or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof of the compound as shown in formula (II),

[0034]

[0035] wherein each --- is independently a single bond or a double bond; n is 1 or 2, and R has the meaning described in the present invention.

[0036] In some embodiments of the present invention, the compound of the present invention is one of the following compounds or its stereoisomers, tautomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts or prodrugs:

[0037]

[0038]

[0039] In yet another aspect, the present invention relates to a pharmaceutical composition comprising the compound of the present invention, and optionally, further comprising a pharmaceutically acceptable excipient or any combination thereof.

[0040] In some embodiments of the composition of the present invention, the pharmaceutical composition further comprises other antiviral drugs, and the other antiviral drugs are enstevir, remdesivir, favipiravir, namatevir, ribavirin, monoupivir, baricitinib, monoupivir, paxlovid, azithromycin, camostat, nafamostat, ritonavir, lopinavir, nitazoxanide, chloroquine, hydroxychloroquine, darunavir, arbidol, resveratrol, interferon α, interferon β, chlorpromazine, imatinib, oseltamivir, loperamide, darunavir, nelfinavir, Jinhua Qinggan Granules, Shufeng Jiedu Capsules, Lianhua Qingwen Capsules or any combination thereof.

[0041] In another aspect, the present invention relates to the use of the compound or the pharmaceutical composition of the present invention in the preparation of a medicament for preventing, treating, curing or alleviating a disease or symptom caused by a coronavirus, wherein the coronavirus is HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV or SARS-CoV-2.

[0042] In another aspect, the present invention also relates to a pharmaceutical composition comprising the compound of the present invention.

[0043] In some embodiments, the pharmaceutical composition described in the present invention further comprises a pharmaceutically acceptable excipient or a combination of such excipients.

[0044] In some embodiments, the pharmaceutical composition of the present invention further comprises other antiviral drugs.

[0045] In some embodiments, the other antiviral drugs described in the present invention are remdesivir, favipiravir, namatevir, ribavirin, monoupivir, baricitinib, monoupivir, paxlovid, azithromycin, camostat, nafamostat, ritonavir, lopinavir, nitazoxanide, chloroquine, hydroxychloroquine, darunavir, arbidol, resveratrol, interferon α, interferon β, chlorpromazine, imatinib, oseltamivir, loperamide, darunavir, nelfinavir, Jinhua Qinggan Granules, Shufeng Jiedu Capsules, Lianhua Qingwen Capsules, or any combination thereof.

[0046] On the other hand, the present invention relates to the use of the compound described in the present invention or the pharmaceutical composition described in the present invention in the preparation of a medicament for preventing, treating, curing or alleviating coronavirus.

[0047] In some embodiments, the coronavirus described in the present invention is HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV or SARS-CoV-2.

[0048] In some embodiments, the present invention relates to the use of a compound represented by formula (I) or (II) or a pharmaceutical composition comprising a compound represented by formula (I) or (II) in the preparation of a medicament for preventing, treating or alleviating diseases or symptoms caused by HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV or SARS-CoV-2.

[0049] In some embodiments, the present invention relates to a method for preventing, treating or alleviating a disease or symptom caused by HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV or SARS-CoV-2, the method comprising administering to a patient a pharmaceutically acceptable effective dose of a compound represented by formula (I) or (II) of the present invention or a pharmaceutical composition comprising a compound represented by formula (I) or (II).

[0050] The foregoing description only summarizes certain aspects of the present invention, but is not limited to these aspects. These aspects and other aspects will be described in more detail and complete below.

[0051] Detailed description of the present invention

[0052] Definitions and general terms

[0053] The present invention will list the literature corresponding to the determined specific content in detail, and the embodiments are accompanied by diagrams of structural formulas and chemical formulas. The present invention is expected to cover all options, variants and equivalents, which may be included in the existing invention field as defined in the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which can be applied to the practice of the present invention. The present invention is by no means limited to the description of methods and materials. There are many documents and similar materials that are different from or conflict with the present application, including but not limited to the definition of terms, the usage of terms, the technology described, or the scope described in the present application.

[0054] The following definitions shall apply to the present invention unless otherwise indicated. For the purposes of the present invention, the chemical elements are those according to the Periodic Table of the Elements, CAS version and the Handbook of Chemicals, 75, thEd, 1994. In addition, the general principles of organic chemistry are defined in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, and all contents are incorporated by reference.

[0055] As described herein, the compounds of the present invention may optionally be substituted with one or more substituents, such as the general formula compounds above, or as specific examples in the embodiments, subclasses, and classes of compounds encompassed by the present invention. In general, the term "substituted" means that one or more hydrogen atoms in a given structure are replaced by a specific substituent. Unless otherwise indicated, an optional substituted group may have a substituent substituted at each substitutable position of the group. When more than one position in a given structural formula can be substituted with one or more substituents selected from a specific group, the substituents may be substituted at each position in the same or different manner.

[0056] In various parts of this specification, the substituents of the compounds disclosed in the present invention are disclosed according to group types or ranges. It is particularly pointed out that the present invention includes each independent subcombination of the individual members of these group types and ranges. For example, the term "C 1-6 "Alkyl" specifically refers to methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl and C6 alkyl as disclosed independently.

[0057] The term "alkyl" used in the present invention includes 1-20 carbon atoms saturated straight or branched monovalent hydrocarbon groups, wherein the alkyl group can be independently optionally substituted with one or more substituents described in the present invention. In some embodiments, the alkyl group contains 1-12 carbon atoms, in other embodiments, the alkyl group contains 1-10 carbon atoms, in other embodiments, the alkyl group contains 1-8 carbon atoms, in other embodiments, the alkyl group contains 1-6 carbon atoms, in other embodiments, the alkyl group contains 1-4 carbon atoms, and in other embodiments, the alkyl group contains 1-3 carbon atoms. Further examples of alkyl groups include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), n-propyl (n-Pr, -CH2CH2CH3), isopropyl (i-Pr, -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), 2-methylpropyl or isobutyl (i-Bu, -CH2CH(CH3)2), 1-methylpropyl or sec-butyl (s-Bu, -CH(CH3)CH 2CH3), tert-butyl (t-Bu, -C(CH3)3), n-pentyl (-CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3) 2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), n-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)C H2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), n-heptyl, n-octyl, and the like.

[0058] The term "alkylene" means a saturated divalent or polyvalent hydrocarbon radical derived from a saturated straight or branched hydrocarbon radical by removing two or more hydrogen atoms. Unless otherwise specified, an alkylene group contains 1-12 carbon atoms. In some embodiments, an alkylene group contains 4-12 carbon atoms; in some embodiments, an alkylene group contains 6-12 carbon atoms; in some embodiments, an alkylene group contains 8-11 carbon atoms; in some embodiments, an alkylene group contains 9-10 carbon atoms; in some embodiments, an alkylene group contains 2-8 carbon atoms; in some embodiments, an alkylene group contains 1-6 carbon atoms; in some embodiments, an alkylene group contains 2-8 carbon atoms; in some embodiments, an alkylene group contains 3-7 carbon atoms; in some embodiments, an alkylene group contains 4-6 carbon atoms; in other embodiments, an alkylene group contains 1-4 carbon atoms; in still other embodiments, an alkylene group contains 1-3 carbon atoms; in still other embodiments, an alkylene group contains 1-2 carbon atoms. Examples of alkylene groups include, but are not limited to, methylene (-CH2-), ethylene (-CH2CH2-), isopropylene (-CH(CH3)CH2-), -(CH2)9-, -(CH2) 10 -etc.

[0059] The terms "hydroxyalkyl" and "hydroxyalkoxy" respectively represent an alkyl or alkoxy group, as the case may be, substituted with one or more hydroxy groups, wherein "hydroxyalkyl" and "hydroxyalkyl" are used interchangeably, such examples include, but are not limited to, hydroxymethyl (-CH2OH), hydroxyethyl (-CH2CH2OH, -CH(OH)CH3), hydroxypropyl (-CH2CH2CH2OH, -CH2CH(OH)CH3, -CH(OH)CH2CH3), hydroxymethoxy (-OCH2OH), and the like.

[0060] The term "haloalkyl", "haloalkenyl" or "haloalkoxy" means that an alkyl, alkenyl or alkoxy group is substituted with one or more halogen atoms. Examples include, but are not limited to, difluoroethyl (-CH2CHF2, -CF2CH3, -CHFCH2F), trifluoroethyl (-CH2CF3, -CF2CH2F, -CFHCHF2), trifluoromethyl (-CF3), trifluoromethoxy (-OCF3), etc.

[0061] The term "alkoxy" means an alkyl group attached to the rest of the molecule via an oxygen atom, wherein the alkyl group has the meaning as described herein. Unless otherwise specified, the alkoxy group contains 1-12 carbon atoms. In some embodiments, the alkoxy group contains 1-8 carbon atoms; in other embodiments, the alkoxy group contains 1-6 carbon atoms; in other embodiments, the alkoxy group contains 1-4 carbon atoms; in yet other embodiments, the alkoxy group contains 1-3 carbon atoms. The alkoxy group may be optionally substituted with one or more substituents described herein.

[0062] Examples of alkoxy groups include, but are not limited to, methoxy (MeO, -OCH3), ethoxy (EtO, -OCH2CH3), 1-propoxy (n-PrO, n-propoxy, -OCH2CH2CH3), 2-propoxy (i-PrO, i-propoxy, -OCH(CH3)2), 1-butoxy (n-BuO, n-butoxy, -OCH2CH2CH2CH3), 2-methyl-l-propoxy (i-BuO, i-butoxy, -OCH2CH(CH3)2), 2-butoxy (s-BuO, s-butoxy, -OCH(CH3)CH2CH3), 2-methyl-2- Propoxy (t-BuO, t-butoxy, -OC(CH3)3), 1-pentyloxy (n-pentyloxy, -OCH2CH2CH2CH2CH3), 2-pentyloxy (-OCH(CH3)CH2CH2CH3), 3-pentyloxy (-OCH(CH2CH3)2), 2-methyl-2-butoxy (-OC(CH3)2CH2CH3), 3-methyl-2-butoxy (-OCH(CH3)CH(CH3)2), 3-methyl-l-butoxy (-OCH2CH2CH(CH3)2), 2-methyl-l-butoxy (-OCH2CH(CH3)CH2CH3) and the like.

[0063] The term "alkenyl" means a straight or branched monovalent hydrocarbon radical containing 2-12 carbon atoms, or 4-12 carbon atoms, or 6-12 carbon atoms, or 8-11 carbon atoms, or 9-10 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, wherein at least one position of CC is sp 2 Double bond, wherein the alkenyl group can be independently unsubstituted or substituted with one or more substituents described in the present invention, including "cis", "trans" or "Z", "E" isomers, specific examples of which include, but are not limited to, vinyl (-CH=CH2), propenyl (-CH=CHCH3), allyl (-CH2CH=CH2) and the like, wherein the alkenyl group can be independently unsubstituted or substituted with one or more substituents described in the present invention.

[0064] The term "alkenylene" refers to a divalent or multivalent alkenyl group derived from a straight or branched alkenyl group by removing two or more hydrogen atoms. Unless otherwise specified, an alkenylene group contains 2-12 carbon atoms. In some embodiments, an alkenylene group contains 2-6 carbon atoms; in some embodiments, an alkenylene group contains 2-8 carbon atoms; in some embodiments, an alkenylene group contains 3-7 carbon atoms; in some embodiments, an alkenylene group contains 4-6 carbon atoms. Examples of alkenylene groups include, but are not limited to -(CH2)6-CH=CHCH2-, -(CH2)7-CH=CHCH2-, -(CH2)5-CH=CHCH2CH2-, -(CH2)-CH2CH=CHCH2-, -(CH2)6-CH=CHCH2-, -(CH2)7-CH=CHCH2-, -(CH2)5-CH=CHCH2CH2-, and -(CH2)-CH2CH=CHCH2-, and the like.

[0065] The term "cycloalkyl" refers to a monovalent or polyvalent saturated monocyclic, bicyclic or tricyclic ring system containing 3 to 12 carbon atoms. In one embodiment, the cycloalkyl contains 3 to 12 carbon atoms; in another embodiment, the cycloalkyl contains 3 to 8 carbon atoms; in another embodiment, the cycloalkyl contains 3 to 7 carbon atoms; in yet other embodiments, the cycloalkyl contains 3 to 6 carbon atoms. Examples of cycloalkyl further include, but are in no way limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0066] The term "consisting of M-M1 ring atoms" means that the cyclic group is composed of M-M1 ring atoms, and the ring atoms include carbon atoms and / or heteroatoms such as O, N, S, and P. For example, "a monocyclic heteroaryl group consisting of 5-6 ring atoms" represents a monocyclic heteroaryl group consisting of 5 or 6 ring atoms; for example, "a bicyclic heteroaryl group consisting of 9-10 ring atoms" represents a bicyclic heteroaryl group consisting of 5 or 6 ring atoms.

[0067] The term "n ring atoms", where n is an integer, typically describes the number of ring atoms in a molecule, where the number of ring atoms is n. For example, piperidinyl is a heterocyclyl group consisting of 6 ring atoms.

[0068] The term "heterocyclyl" refers to a non-aromatic, saturated or partially unsaturated monocyclic, bicyclic or tricyclic ring system containing 3-12 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur or oxygen atoms. Wherein, the heterocyclyl group may be optionally substituted by one or more substituents described in the present invention. Unless otherwise specified, the heterocyclyl group may be a carbon group or a nitrogen group, and the -CH2- group may be optionally replaced by -C(=O)- or -C(=S)-. The sulfur atom of the ring may be optionally oxidized to S-oxide. The nitrogen atom of the ring may be optionally oxidized to N-oxide. In some embodiments, the heterocyclyl group is a heterocyclyl group consisting of 3-12 ring atoms. In some embodiments, the heterocyclic group is a heterocyclic group consisting of 3-10 ring atoms, in some embodiments, the heterocyclic group is a heterocyclic group consisting of 5-10 ring atoms, in some embodiments, the heterocyclic group is a heterocyclic group consisting of 3-6 ring atoms, in some embodiments, the heterocyclic group is a heterocyclic group consisting of 4-6 ring atoms, in some embodiments, the heterocyclic group is a heterocyclic group consisting of 5-6 ring atoms, in other embodiments, the heterocyclic group is a heterocyclic group consisting of 4 atoms, which refers to a monovalent or polyvalent, saturated or partially unsaturated, non-aromatic monocyclic ring containing 4 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. In other embodiments, the heterocyclic group is a heterocyclic group consisting of 5 atoms, which refers to a monovalent or polyvalent, saturated or partially unsaturated, non-aromatic monocyclic ring containing 5 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. In other embodiments, the heterocyclic group is a 6-atom heterocyclic group, which refers to a monovalent or polyvalent, saturated or partially unsaturated, non-aromatic monocyclic ring containing 6 ring atoms, wherein at least one ring atom is selected from nitrogen, sulfur and oxygen atoms. "Heterocyclic group" also includes groups formed by condensing a heterocyclic group with a saturated or partially unsaturated ring or a heterocyclic ring. Examples of heterocyclic rings include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, piperazinyl, homopiperazinyl, azetidinyl, oxetanyl, thiidine, homopiperidinyl, glycidyl, azepanyl, oxetanyl, thiidine, oxazepinyl, diazepinyl, thiazepinyl, 2-pyrrolinyl, 3- pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolane, pyrazolinyl, dithianyl, dithiolanyl, dihydrothienyl, pyrazolidinylimidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, 3H-indolylquinolizinyl and N-pyridylurea.Examples of heterocyclic groups also include 1,1-dioxothiomorpholinyl, examples in which the carbon atoms on the ring are replaced by oxo (=O) include, but are not limited to, pyrimidinedione, 1,2,4-thiadiazole-5(4H)-one, 1,2,4-oxadiazole-5(4H)-one, 1H-1,2,4-triazole-5(4H)-one, and the like, examples in which the carbon atoms on the ring are replaced by a =S group include, but are not limited to, 1,2,4-oxadiazole-5(4H)-thione, 1,3,4-oxadiazole-2(3H)-thione, and the like.

[0069] The term "heteroatom" means one or more O, S, N, P and Si, including any oxidation state of N, S and P; in the form of primary, secondary, tertiary amines and quaternary ammonium salts; or in the form of a nitrogen atom in a heterocyclic ring in which the hydrogen is substituted, for example, N (such as N in 3,4-dihydro-2H-pyrrolyl), NH (such as NH in pyrrolidinyl) or NR (such as NR in N-substituted pyrrolidinyl, wherein R represents a substituent as described in the present invention).

[0070] The term "halogen" or "halogen atom" refers to F, Cl, Br or I.

[0071] As used herein, the term "unsaturated" means that the moiety contains one or more degrees of unsaturation.

[0072] The term "aryl" refers to a monocyclic, bicyclic or tricyclic carbon ring system containing 6-14 ring atoms, or 6-12 ring atoms, or 6-10 ring atoms, wherein at least one ring system is aromatic, wherein each ring system comprises a ring consisting of 3-7 atoms and has one or more points of attachment to the rest of the molecule. The term "aryl" can be used interchangeably with the term "aromatic ring". Examples of aryl groups can include phenyl, naphthyl and anthracenyl. The aryl groups can be independently and optionally substituted with one or more substituents described herein.

[0073] The term "heteroaryl" refers to monocyclic, bicyclic and tricyclic ring systems containing 5-12 ring atoms, or 5-10 ring atoms, or 9-10 ring atoms, or 9 ring atoms, or 5-6 ring atoms, wherein at least one ring is aromatic, and at least one aromatic ring contains one or more heteroatoms, wherein each ring system contains 5-7 atoms and has one or more points of attachment to the rest of the molecule. The term "heteroaryl" can be used interchangeably with the terms "aromatic heterocycle", "heteroaromatic ring" or "heteroaromatic compound". In some embodiments, heteroaryl is a heteroaryl composed of 5-12 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, sulfur and oxygen. In other embodiments, heteroaryl is a heteroaryl composed of 5-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, sulfur and oxygen. In other embodiments, heteroaryl is a bicyclic heteroaryl consisting of 9-10 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, sulfur and oxygen. In other embodiments, heteroaryl is a heteroaryl consisting of 9 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, sulfur and oxygen. In other embodiments, heteroaryl is a monocyclic heteroaryl consisting of 5-6 atoms containing 1, 2, 3 or 4 heteroatoms independently selected from nitrogen, sulfur and oxygen. And the heteroaryl can be substituted or unsubstituted, and the aryl group can be independently and optionally substituted with one or more substituents described in the present invention.

[0074] Examples of heteroaromatic rings include, but are not limited to, the following monocyclic heteroaryl groups: 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl, pyridazinyl (such as 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (such as 5-tetrazolyl), triazolyl (such as 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyranyl, pyrazolyl (such as 2-pyrazolyl), isothiazolyl, The following bicyclic heteroaryl groups are also included, but are by no means limited to these bicyclic groups: benzothiazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl, indolyl (such as 2-indolyl), purinyl, quinolyl (such as 2-quinolyl, 3-quinolyl, 4-quinolyl), isoquinolyl (such as 1-isoquinolyl, 3-isoquinolyl or 4-isoquinolyl), benzopyrazolyl, benzopyrrolyl, benzoxazolyl, benzisoxazolyl, etc.

[0075] In addition, it should be noted that, unless explicitly stated otherwise, the description methods used throughout this article, "each... and... are independently", "... and... are each independently", and "... and... are respectively independently" are interchangeable and should be understood in a broad sense, which can mean that in different groups, the specific options expressed by the same symbols do not affect each other, or that in the same group, the specific options expressed by the same symbols do not affect each other.

[0076] The term "room temperature" refers to 10°C to 40°C, in some embodiments, "room temperature" refers to 10°C to 30°C; in some embodiments, "room temperature" refers to 20°C to 30°C.

[0077] Unless otherwise indicated, the structural formulas described in the present invention include all isomeric forms (such as enantiomers, diastereomers, and geometric isomers (or conformational isomers): for example, R, S configurations containing asymmetric centers, (Z), (E) isomers of double bonds, and (Z), (E) conformational isomers. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers, or geometric isomers (or conformational isomers) are all within the scope of the present invention.

[0078] The term "prodrug" as used in the present invention refers to a compound that is converted into a compound represented by formula (I) or (II) in vivo. Such conversion is affected by the prodrug being hydrolyzed in the blood or being converted into the parent structure by enzymes in the blood or tissues. The prodrug compound of the present invention may be an ester. In the prior invention, esters that can be used as prodrugs include phenyl esters, aliphatic (C 1-24 ) esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters. For example, a compound of the present invention contains a hydroxyl group, which can be acylated to obtain a prodrug form of the compound. Other prodrug forms include phosphates, such as these phosphate compounds obtained by phosphorylation of the hydroxyl group on the parent. For a complete discussion of prodrugs, please refer to the following literature: T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series, Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, J. Rautio et al, Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270, and SJ Hecker et al, Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51, 2328-2345.

[0079] Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are included within the scope of the present invention. In addition, unless otherwise indicated, the structural formulas of the compounds described in the present invention include enriched isotopes of one or more different atoms.

[0080] "Metabolite" refers to a product obtained by the metabolism of a specific compound or salt thereof in vivo. The metabolites of a compound can be identified by techniques known in the art, and their activity can be characterized by experimental methods as described herein. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic cleavage, etc. Accordingly, the present invention includes metabolites of compounds, including metabolites produced by contacting a compound of the present invention with a mammal for a period of time.

[0081] The definitions and conventions of stereochemistry used in this invention are generally based on the following references: SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomers. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, constitute a part of the present invention. Many organic compounds exist in optically active forms, that is, they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefixes D, L or R, S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes d, l or (+), (-) are used to name the sign of rotation of plane polarized light of a compound. (-) or l means that the compound is levorotatory, and the prefix (+) or d means that the compound is dextrorotatory. The chemical structures of these stereoisomers are identical, but their stereostructures are different. Specific stereoisomers can be enantiomers, and a mixture of isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may result in no stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers that lacks optical activity.

[0082] The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can be interconverted through a low energy barrier. For example, proton tautomers (i.e., prototropic tautomers) include interconversions through proton migration, such as keto-enol and imine-enamine isomerization. Valence (chemical valence) tautomers include interconversions by reorganization of bonding electrons. Unless otherwise indicated, all tautomeric forms of the compounds of the invention are within the scope of the invention.

[0083] One or more hydrogen atoms, carbon atoms or other atoms of the compounds of formula (I) or (II) of the present invention may be replaced by isotopes of hydrogen atoms, carbon atoms or other atoms. The compounds of formula (I) or (II) include all radioactive labels of the compounds of formula (I) or (II). Such "radioactive labeling", "radioactive labels" and the like of the compounds of formula (I) or (II) are respectively included in the present invention and can be used for pharmacokinetic studies and studies in binding assays and / or as diagnostic tools. Examples of isotopes that can be incorporated into the compounds of formula (I) or (II) of the present invention include 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36 Cl, such hydrogen atom, carbon atom, nitrogen atom, oxygen atom, phosphorus atom, sulfur atom, fluorine atom and chlorine atom. The radiolabeled compound of the present invention can be prepared by a method known in the art.

[0084] The "pharmaceutically acceptable salt" used in the present invention refers to the organic salt and inorganic salt of the compound of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in the literature: SM Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19, 1977. Pharmaceutically acceptable salts formed by non-toxic acids include, but are not limited to, inorganic acid salts formed by reaction with amino groups, such as hydrochlorides, hydrobromides, phosphates, sulfates, perchlorates, and organic acid salts such as acetates, oxalates, maleates, tartrates, citrates, succinates, malonates, or other methods described in books and literature, such as ion exchange methods, to obtain these salts. Other pharmaceutically acceptable salts include adipate, malate, 2-hydroxypropionate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C 1-4 The present invention also contemplates quaternary ammonium salts formed by any compound containing a N group. Water-soluble or oil-soluble or dispersible products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium salts and amine cations formed by counter ions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C 1-8 Sulfonates and aromatic sulfonates.

[0085] The "solvate" of the present invention refers to an association formed by one or more solvent molecules and the compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol. The term "hydrate" refers to an association formed when the solvent molecule is water.

[0086] Detailed description of the compounds of the present invention

[0087] The compounds involved in the present invention and their pharmaceutically acceptable compositions can effectively inhibit diseases or symptomatic infections caused by SARS-CoV-2.

[0088] Unless otherwise indicated, all stereoisomers, geometric isomers, tautomers, N-oxides, hydrates, solvates, metabolites, salts and pharmaceutically acceptable prodrugs of the compounds used in the pharmaceutical use of the present invention are within the scope of the present invention.

[0089] In particular, the salts are pharmaceutically acceptable salts. The term "pharmaceutically acceptable" includes that the substance or composition must be suitable chemically and toxicologically with the other ingredients that make up the formulation and with the mammal to be treated.

[0090] The salts of the compounds of the present invention also include salts of intermediates used in the preparation or purification of the compounds of formula (I) or (II) or separated enantiomers of the compounds of formula (I) or (II), but are not necessarily pharmaceutically acceptable salts.

[0091] If the compound of the present invention is basic, the desired salt can be prepared by any suitable method provided in the literature, for example, using inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid, etc. Or using organic acids such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, malic acid, 2-hydroxypropionic acid, citric acid, oxalic acid, glycolic acid and salicylic acid; pyranose acids such as glucuronic acid and galacturonic acid; α-hydroxy acids such as citric acid and tartaric acid; amino acids such as aspartic acid and glutamic acid; aromatic acids such as benzoic acid and cinnamic acid; sulfonic acids such as p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, ethanesulfonic acid, trifluoromethanesulfonic acid, etc. or combinations thereof.

[0092] If the compound of the present invention is acidic, the desired salt can be prepared by a suitable method, for example, using an inorganic base or an organic base, such as ammonia (primary, secondary, tertiary), an alkali metal hydroxide, ammonium, N + (R 14 )4 and alkaline earth metal hydroxides, and the like. Suitable salts include, but are not limited to, organic salts derived from amino acids, such as glycine and arginine, amines, such as primary, secondary and tertiary amines, N + (R 14 )4 salts, such as R 14 It is H, C 1-4 Alkyl, C 6-10 Aryl, C 6-10 Aryl C 1-4alkyl, etc., and cyclic amines such as piperidine, morpholine and piperazine, etc., and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium. Also included are appropriate, non-toxic ammonium, quaternary ammonium salts and amine cations that counteract the formation of counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C 1-8 Sulfonates and aromatic sulfonates.

[0093] Pharmaceutical compositions, formulations, administration and uses of the compounds and pharmaceutical compositions of the present invention

[0094] The pharmaceutical composition of the present invention comprises a compound of formula (I) or (II), a compound listed in the present invention, or an example compound, and a pharmaceutically acceptable excipient. The pharmaceutical composition of the present invention can effectively inhibit coronaviruses and is suitable for the treatment of coronavirus infections, especially for diseases or symptoms caused by HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV or SARS-CoV-2.

[0095] The compounds described in the present invention exist in free form, or are suitable as pharmaceutically acceptable derivatives. According to the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable prodrugs, salts, esters, salts of esters, or any other adducts or derivatives that can be directly or indirectly administered to patients as needed, compounds described in other aspects of the present invention, their metabolites or their residues.

[0096] As described in the present invention, the pharmaceutical composition of the present invention comprises any one of the compounds represented by formula (I) or (Ia), (II-a), (II-b), (III-a) or (III-b) of the present invention, and further comprises pharmaceutically acceptable excipients, such as those used in the present invention, including any solvent, solid excipient, diluent, binder, disintegrant, or other liquid excipient, dispersant, flavoring agent or suspending agent, surfactant, isotonic agent, thickener, emulsifier, preservative, solid binder or lubricant, etc., suitable for a specific target dosage form. As described in the following document: In Remington: The Science and Practice of Pharmacy, 21st edition, 2005, ed. DB Troy, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JC Boylan, 1988-1999, Marcel Dekker, New York, the content of the document is integrated here, showing that different adjuvants can be applied to the preparation of pharmaceutically acceptable compositions and their known preparation methods. Except the scope that any conventional adjuvant is incompatible with the compound of the present invention, for example any adverse biological effect produced or the interaction produced in a harmful manner with any other component of pharmaceutically acceptable compositions, their use is also the scope considered by the present invention.

[0097] Substances that can be used as pharmaceutically acceptable excipients include, but are not limited to, ion exchangers; aluminum; aluminum stearate; lecithin; serum proteins, such as human serum albumin; buffer substances such as phosphates; glycine; sorbic acid; potassium sorbate; partial glyceride mixtures of saturated vegetable fatty acids; water; salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts; colloidal silicon; magnesium trisilicate; polyvinyl pyrrolidone; polyacrylates; waxes; polyethylene-polyoxypropylene-blocking polymers; lanolin; sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as carboxymethyl cellulose sodium cellulose, ethylcellulose and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycol compounds such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; phosphate buffer solution; and other non-toxic suitable lubricants such as sodium lauryl sulfate and magnesium stearate; colorants; release agents; coatings; sweeteners; flavorings; fragrances; preservatives and antioxidants.

[0098] The pharmaceutical composition provided by the present invention also contains other antiviral drugs.

[0099] The other antiviral drugs are remdesivir, favipiravir, ribavirin, camostat, nafamostat, ritonavir, lopinavir, nitazoxanide, chloroquine, hydroxychloroquine, darunavir, arbidol, resveratrol, interferon α, interferon β, chlorpromazine, imatinib, oseltamivir, loperamide, darunavir, nelfinavir, Jinhua Qinggan Granules, Shufeng Jiedu Capsules, Lianhua Qingwen Capsules, etc., or any combination thereof.

[0100] The compound or pharmaceutical composition of the present invention can be administered in any of the following ways: oral administration, spray inhalation, topical administration, rectal administration, nasal administration, topical administration, vaginal administration, parenteral administration such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal, or intracranial injection or infusion, or medication via an explanted reservoir. The preferred mode is oral administration, intramuscular injection, intraperitoneal administration or intravenous injection.

[0101] The compounds of the present invention or pharmaceutically acceptable compositions thereof may be administered in a unit dosage form. The dosage form may be a liquid dosage form or a solid dosage form. The liquid dosage form may be a true solution, a colloid, a microparticle dosage form, or a suspension dosage form. Other dosage forms include tablets, capsules, dripping pills, aerosols, pills, powders, solutions, suspensions, emulsions, granules, suppositories, freeze-dried powder injections, inclusion compounds, implants, patches, liniments, etc.

[0102] Tablets and capsules for oral administration may contain excipients such as binders, such as syrup, gum arabic, sorbitol, tragacanth, or polyvinyl pyrrolidone; fillers, such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, aminoacetic acid; lubricants, such as magnesium stearate, talc, polyethylene glycol, silica; disintegrants, such as potato starch; or acceptable wetting agents such as sodium lauryl sulfate. Tablets may be coated by methods known in the pharmaceutical industry.

[0103] Oral liquid can be made into suspension, solution, emulsion, syrup or elixir of hydrated oil, or it can be made into dry product, and water or other suitable medium is supplemented before use. Such liquid preparations can contain conventional additives, such as suspending agent, sorbitol, methyl cellulose, glucose syrup, gel, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel, hydrogenated edible oil, emulsifier, such as lecithin, sorbitan monooleate, gum arabic; or non-aqueous excipients (may contain edible oil), such as almond oil, oil such as glycerol, ethylene glycol, or ethanol; preservative, such as methyl or propyl parahydroxybenzoate, sorbic acid. Flavoring agent or coloring agent can be added as needed.

[0104] Suppositories may contain conventional suppository bases such as cocoa butter or other glycerides.

[0105] For extragastric administration, liquid dosage forms are usually made of the compound and a sterile excipient. The excipient is preferably water. Depending on the selected excipient and the drug concentration, the compound can be dissolved in the excipient or made into a suspension solution. When making an injection solution, the compound is first dissolved in water, filtered and sterilized, and then filled into a sealed bottle or ampoule.

[0106] When applied topically to the skin, the compounds of the present invention can be made into the form of suitable ointments, lotions, or creams, in which the active ingredient is suspended or dissolved in one or more excipients, wherein the excipients that can be used for ointment preparations include but are not limited to: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; the excipients that can be used for lotions and creams include but are not limited to: mineral oil, sorbitan monostearate, Tween 60, hexadecyl esters wax, hexadecene aromatic alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0107] In general, it has proven to be advantageous that the total amount of the active compound of the invention administered per 24 hours is about 0.0005-500 mg / kg body weight, preferably 0.5-100 mg / kg body weight, if appropriate, in a plurality of single doses to achieve the desired effect, both in human and veterinary medicine. The amount of active compound contained in a single dose is preferably about 1-80 mg / kg body weight, more preferably 1-50 mg / kg body weight, but it may not be in accordance with the above dosage, i.e., it depends on the type and body weight of the treated subject, the nature and severity of the disease, the type of preparation and the mode of administration of the drug, as well as the administration cycle or time interval.

[0108] Coronavirus infected drugs can be administered separately from the composition comprising the compounds of the present invention as part of a multiple dosing regimen. Alternatively, those drugs can be part of a single dosage form, mixed with the compounds of the present invention to form a single composition. If the administration is part of a multiple dosing regimen, the two active agents can be delivered to each other simultaneously, continuously or over a period of time, thereby obtaining the target agent activity.

[0109] The amount of the compounds and compositions (those containing a composition such as described herein) that can be combined with the excipient materials to produce a single dosage form will vary depending on the indication and the particular mode of administration. Normally, the amount of the composition of the present invention will not exceed the amount normally administered for a composition containing the agent as the only active agent. On the other hand, the amount of the presently disclosed composition ranges from about 50% to 100% of the normal amount of existing compositions containing the agent as the only active therapeutic agent. In those compositions containing the composition will act synergistically with the compounds of the present invention.

[0110] General Synthesis Methods of Compounds Described in the Present Invention

[0111] Generally, the compounds of the present invention can be prepared by the methods described herein, unless otherwise specified, wherein the substituents are defined as shown in formula (I) or (II). The following reaction schemes and examples are provided to further illustrate the present invention.

[0112] In the examples described below, all temperatures are in degrees Celsius (°C) unless otherwise indicated. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, Arco Chemical Company and Alfa Chemical Company and were used without further purification unless otherwise indicated. Common reagents were purchased from Shantou Xilong Chemical Factory, Guangdong Guanghua Chemical Reagent Factory, Guangzhou Chemical Reagent Factory, Tianjin Haoyuyu Chemical Co., Ltd., Qingdao Tenglong Chemical Reagent Co., Ltd., and Qingdao Ocean Chemical Factory.

[0113] The chromatographic column used was a silica gel column, and the silica gel (200-300 mesh) was purchased from Qingdao Ocean Chemical Plant. The NMR spectra were CDC13, DMSO-d6, CD3OD or acetone-d6 as solvents (reported in ppm), and TMS (0 ppm) or chloroform (7.26 ppm) was used as reference standards. When multiple peaks appeared, the following abbreviations were used: s (singlet), d (doublet), t (triplet), m (multiplet), br (broadened), dd (doublet of doublets), dt (doublet of triplets), br.s (broadened singlet), q (quartet),. The coupling constant J is expressed in Hertz (Hz).

[0114] Low-resolution mass spectrometry (MS) data were determined by an Agilent 6320 series LC-MS spectrometer equipped with a G1312A binary pump and a G1316A TCC (column temperature was maintained at 30 °C). A G1329A autosampler and a G1315B DAD detector were used for analysis, and an ESI source was applied to the LC-MS spectrometer.

[0115] Low-resolution mass spectrometry (MS) data were determined by an Agilent 6120 series LC-MS spectrometer equipped with a G1311A quaternary pump and a G1316A TCC (column temperature was maintained at 30 °C). A G1329A autosampler and a G1315D DAD detector were used for analysis, and an ESI source was applied to the LC-MS spectrometer.

[0116] Both spectrometers were equipped with an Agilent Zorbax SB-C18 column with a size of 2.1×30 mm, 5 μm. The injection volume was determined by the sample concentration; the flow rate was 0.6 mL / min; the HPLC peak was recorded and read at UV-Vis wavelengths at 210 nm and 254 nm. The mobile phase was a 0.1% formic acid acetonitrile solution (phase A) and a 0.1% formic acid ultrapure aqueous solution (phase B). The purity of the compound was evaluated by an Agilent 1100 series high performance liquid chromatograph (HPLC).

[0117] The following abbreviations are used throughout this invention:

[0118]

[0119] DETAILED DESCRIPTION

[0120] The following examples are used to illustrate the present invention but are not intended to limit the scope of the present invention.

[0121] Preparation Example

[0122] In the following preparation examples, the inventors describe in detail the preparation process of the compounds of the present invention by taking some of the compounds of the present invention as examples.

[0123] Example 1: Synthesis of compound (1)

[0124]

[0125] Step 1: Synthesis of compound 1-2

[0126] Compound 1-1 (2.19 g, 8.07 mmol) was dissolved in DMF (15 mL), and cesium carbonate (1.31 g, 4.04 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Methyl iodide (1.00 mL, 16.14 mmol) was added, and the mixture was stirred at room temperature for 3.5 h. After the reaction was completed, water (100 mL) was added to the reaction solution, and EA (50 mL × 3) was used for extraction. The organic phases were combined, washed with saturated saline solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 1 / 9) to obtain the title compound 1-2 (2.17 g, 94.22%), a colorless oily liquid. MS (ESI, pos.ion) m / z: 186.2 [M + H-Boc] + .

[0127] Step 2: Synthesis of Compound 1-3

[0128] Compound 1-2 (2.17 g, 7.60 mmol) was dissolved in 1,4-dioxane solution of HCl (19 mL, 76 mmol, 4 M) and stirred at room temperature for 2 h. The reaction was stopped, concentrated under reduced pressure, and deacidified with DCM to obtain the title compound 1-3 (1.68 g, 99.70%) as a white solid, which was used directly in the next step.

[0129] Step 3: Synthesis of Compound 1-4

[0130] Compound 1-3 (1.68 g, 7.58 mmol) was dissolved in DCM (40 mL), and DIPEA (6.28 mL, 37.9 mmol) was added. After stirring at room temperature for 10 minutes, N-Boc-L-tert-leucine (1.75 g, 7.58 mmol) was added, and the temperature was lowered to 0°C, HATU (5.76 g, 15.16 mmol) and HOAT (2.06 g, 15.16 mmol) were added, and stirring was continued for 20 minutes. After the reaction was completed, a mixture of water and HCl solution (1N) was added (V / V=1 / 1, 50 mL), and the resulting mixture was extracted with DCM (50 mL×3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 1 / 9) to obtain the title compound 1-4 (2.60 g, 86.07%) as a colorless liquid. MS (ESI, pos.ion) m / z: 299.4 [M+H-Boc] + .

[0131] Step 4: Synthesis of Compound 1-5

[0132] Compound 1-4 (2.92 g, 7.33 mmol) was dissolved in 1,4-dioxane solution of HCl (18.32 mL, 73.3 mmol, 4 mol / L), stirred at room temperature for 1 h, concentrated under reduced pressure, and deacidified with DCM to obtain the title compound 1-5 (2.44 g, 99.40%) as a white solid, which was used directly in the next step.

[0133] Step 5: Synthesis of Compound 1-6

[0134] Compound 1-5 (2.44 g, 7.29 mmol) was dissolved in DMF (25 mL), potassium carbonate (5.04 g, 36.47 mmol) and sodium iodide (1.09 g, 7.29 mmol) were added, and then 4-bromo-1-butene (1.97 g, 14.58 mmol) was added dropwise. The reaction mixture was heated to 60 °C and stirred for 16.5 h. After the reaction was completed, water (50 mL) was added, and then extracted with EA (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 3 / 17) to obtain the title compound 1-6 (2.17 g, 84.44%), a yellow oily liquid. MS (ESI, pos.ion) m / z: 353.3 [M+H] + .

[0135] Step 6: Synthesis of Compounds 1-7

[0136] Under nitrogen protection, compound 1-6 (0.50 g, 1.42 mmol) was dissolved in DCE (700 mL), ultrasonicated for 5 minutes, heated to 70 ° C, and a solution of Hoveyda-Grubbs catalyst (0.36 g, 0.57 mmol) in DCE (10 mL) was added dropwise under stirring. The reaction mixture was heated to 90 ° C and stirred for 4 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 1 / 4) to obtain the title compound 1-7 (0.18 g, 39.07%), a colorless liquid. MS (ESI, pos.ion) m / z: 325.3 [M+H] + .

[0137] Step 7: Synthesis of Compounds 1-8

[0138] Compound 1-7 (0.18 g, 0.55 mmol) was dissolved in a mixture of THF (4 mL) and methanol (4 mL), lithium hydroxide (0.14 g, 3.31 mmol) and water (0.2 mL) were added in sequence, and the temperature was raised to 60°C and stirred for 1 h. The mixture was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in a mixture of methanol and dichloromethane (MeOH / DCM (V / V) = 1 / 9), and the pH was adjusted to 6 with a cation exchange resin, filtered, and the filtrate was concentrated under reduced pressure to obtain the title compound 1-8 (0.16 g, 92.91%) as a white solid. MS (ESI, pos.ion) m / z: 311.3 [M+H] + .

[0139] Step 8: Synthesis of Compound 1-10

[0140] Compound 1-8 (0.23 g, 0.73 mmol) was dissolved in DMF (5 mL), cooled to 0 ° C, HATU (0.50 g, 1.32 mmol) was added, and then NMM (0.22 mL, 1.98 mmol) was added dropwise, and stirring continued at this temperature for 10 minutes. Finally, compound 1-9 (0.16 g, 0.66 mmol) was added and stirring continued for 50 minutes. After the reaction was completed, water (40 mL) was added, and the reaction mixture was extracted with EA (15 mL × 3). The organic phases were combined, washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the title compound 1-10 (0.32 g, 97.54%) as a light yellow solid. MS (ESI, pos.ion) m / z: 497.3 [M + H] + .

[0141] Step 9: Synthesis of compound (1)

[0142] 2,6-Bistrifluoromethylbenzoic acid (0.99 g, 3.84 mmol) was dissolved in DMF (10 mL), sodium tert-butoxide (0.18 g, 1.87 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Then compound 1-10 (0.32 g, 0.64 mmol) and sodium iodide (0.19 g, 1.27 mmol) were added in sequence, and the reaction mixture was stirred at room temperature overnight. After the reaction was completed, water (40 mL) was added, and the resulting mixture was extracted with EA (20 mL × 3). The organic phases were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative liquid separation (CH3CN / 1‰ FA aqueous solution (V / V) = 3 / 2) to obtain the title compound (1) (46 mg, 9.94%) as a white solid. MS (ESI, pos.ion) m / z: 719.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.77(t,J=7.4Hz,2H),8.53(s,2H),8.26(dd,J=8.1,3.2Hz,2H),8.02(t,J=8.1Hz,1H),7 .67(d,J=7.0Hz,1H),5.82–5.60(m,1H),5.47(dd,J=15.0,7.4Hz,1H),5.32–5.04(m,2H),4.55–4.42(m,2H),3.8 3(d,J=10.7Hz,2H),3.31–2.98(m,3H),2.41(d,J=7.0Hz,1H),2.28(q,J=15.2,12.4Hz,2H),2.20–2.06(m,2H),1 .98(dd,J=18.1,8.4Hz,2H),1.66(q,J=9.3,8.6Hz,4H),1.48(s,2H),1.43–1.34(m,3H),1.06(d,J=10.8Hz,9H).

[0143] Example 2: Synthesis of compound (2)

[0144]

[0145] Step 1: Synthesis of compound 2-2

[0146] Compound 2-1 (2.00 g, 6.99 mmol) was dissolved in methanol (15 mL), transferred to 0 ° C, sodium borohydride (1.62 g, 41.94 mmol) was slowly added, and the reaction mixture was transferred to room temperature and stirred for 12 h. After the reaction was completed, it was concentrated under reduced pressure, diluted with saturated aqueous ammonium chloride solution (30 mL), and then extracted with EA (50 mL × 3), the organic phases were combined, washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (MeOH / DCM (V / V) = 7 / 93) to obtain the title compound 2-2 (1.72 g, 95.26%) as a white solid, which was directly used in the next step.

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

[0148] Compound 2-2 (1.03 g, 3.99 mmol) was dissolved in THF (20 mL), and Dess-Martin periodinane (1.86 g, 4.39 mmol) and sodium bicarbonate (0.34 g, 3.99 mmol) were added and stirred at room temperature for 1 h. After the reaction, sodium thiosulfate pentahydrate solution (50 mL) was added, and then extracted with EA (50 mL × 3), the organic phases were combined, washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (MeOH / DCM (V / V) = 1 / 24) to obtain the title compound 2-3 (701 mg, 68.55%) as a white foamy solid. MS (ESI, pos.ion) m / z: 157.3 [M-Boc+H] + .

[0149] Step 3: Synthesis of compound 2-4

[0150] Under nitrogen protection, sodium hydride (132 mg, 3.29 mmol) was diluted in anhydrous THF (15 mL), and triethyl phosphoacetate (626.82 mg, 2.74 mmol) was slowly added dropwise at -78 ° C. Stirring was continued at this temperature for 30 min, and then a solution of compound 2-3 (701 mg, 2.74 mmol) in anhydrous THF (15 mL) was slowly added dropwise. After the addition was complete, the reaction mixture was heated to -50 ° C and stirred for 2 h. After the reaction was completed, saturated ammonium chloride (5 mL) was added to quench the reaction. The reaction mixture was transferred to room temperature, saturated ammonium chloride (15 mL) was added, and EA (30 mL × 2) was used for extraction. The organic phases were combined, washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (MeOH / DCM (V / V) = 1 / 19) to obtain the title compound 2-4 (325 mg, 36.34%) as a white foamy solid. MS (ESI, pos.ion) m / z: 227.4 [M-Boc+H] + .

[0151] Step 4: Synthesis of Compound 2-5

[0152] Compound 2-4 (240 mg, 0.74 mmol) was dissolved in DCM (1 mL), and a solution of HCl in 1,4-dioxane (1.85 mL, 7.4 mmol, 4 mol / L) was added, and the reaction mixture was stirred at room temperature for 1 h. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain the title compound 2-5 (190 mg, 97.73%) as a white solid, which was used directly in the next step.

[0153] Step 5: Synthesis of compound (2)

[0154] Compound 2-5 (190 mg, 0.72 mmol) and DIPEA (0.61 mL, 3.60 mmol) were dissolved in DCM (5 mL), and then compound 1-8 (80 mg, 0.26 mmol) and HATU (558 mg, 1.44 mmol) were added in sequence at -20 °C, and stirring was continued at this temperature for 15 h. After the reaction was completed, aqueous hydrochloric acid solution (1N) was added to adjust the pH to 5-6. Water (20 mL) was added, and then washed with DCM (20 mL × 2), and the pH of the aqueous phase was adjusted to 7-8 with saturated sodium bicarbonate aqueous solution, and then extracted with EA (30 mL × 2), and the organic phases were combined, and the combined organic phases were washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by preparative liquid phase separation to obtain the title compound (2) (30 mg, 22.44%), a light yellow solid. MS (ESI, pos.ion) m / z: 519.3 [M+H] + . 1H NMR(400MHz, CD3OD)δ6.90(dd,J=15.7,5.1Hz,1H),5.91(dd,J=15.7,1.4Hz,1H),5.71–5.62(m,1H),5.56–5.47(m,1H),4.67–4.59(m,1H) ,4.46(dd,J=11.2,3.4Hz,1H),4.18(q,J=7.1Hz,2H),3.66(s,1H),3.26–3.19(m,1H),2.92–2.85(m,1H),2.60–2.50(m,2H),2.29(ddd,J= 12.0,7.8,6.0Hz,2H),2.19(dd,J=9.4,5.9Hz,1H),2.14–2.06(m,1H),2.07–2.00(m,1H),2.00–1.92(m,1H),1.81(ddt,J=17.7,16.1,11. 0Hz,3H),1.73–1.65(m,1H),1.64–1.54(m,2H),1.52–1.43(m,1H),1.42–1.32(m,4H),1.28(dd,J=13.4,6.4Hz,7H),1.15(d,J=7.2Hz,9H).

[0155] Example 3: Synthesis of compound (3)

[0156]

[0157] Step 1: Synthesis of compound 3-2

[0158] Compound 1-8 (0.14 g, 0.45 mmol) was dissolved in DMF (8 mL), and HOPO (14 mg, 0.13 mmol) and compound 3-1 (111 mg, 0.53 mmol) were added at 0°C, followed by DIPEA (0.22 mL, 1.35 mmol) and EDCI (107 mg, 0.56 mmol). The reaction mixture was transferred to room temperature and stirred for 12 h. After the reaction, water (30 mL) was added, and then extracted with EA (15 mL × 3). The organic phases were combined, washed with saturated brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 1 / 9) to obtain the title compound 3-2 (0.085 g, 40.65%) as a white solid. MS (ESI, pos.ion) m / z: 464.6 [M+H] + .

[0159] Step 2: Synthesis of compound (3)

[0160] Compound 3-2 (85 mg, 0.18 mmol) was dissolved in DCM (6 mL), and Burgess reagent (108 mg, 0.45 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, saturated sodium bicarbonate solution (20 mL) was added, and DCM (10 mL×2) was used for extraction. The organic phases were combined and concentrated under reduced pressure. The residue was separated and purified by preparative liquid phase separation (CH3CN / 1‰ TFA aqueous solution (V / V) = 3 / 7) to obtain the title compound (3) (10 mg, 12.24%) as a white solid. MS (ESI, pos.ion) m / z: 446.3 [M+H] + . 1 H NMR (400MHz, CD3OD) δ5.65(dd,J=14.7,7.4Hz,1H),5.52(dd,J=16.1,8.6Hz,1H),5.02(dd,J=10.1, 5.9Hz,1H),4.43(dd,J=10.7,4.0Hz,1H),3.68(d,J=5.1Hz,1H),3.35(s,1H),3.30–3.21(m,2H),2.9 1(s,1H),2.60–2.47(m,2H),2.26(tdd,J=28.1,19.7,8.8Hz,4H),2.11(s,1H),1.94–1.71(m,4H),1. 69–1.52(m,2H),1.47(d,J=11.5Hz,2H),1.36(d,J=15.5Hz,3H),1.28(d,J=9.2Hz,3H),1.13(s,9H).

[0161] Example 4: Synthesis of Compound (4)

[0162]

[0163] Step 1: Synthesis of compound 4-2

[0164] Compound 4-1 (10.00 g, 65.97 mmol) was dissolved in methanol (40 mL), and thionyl chloride (23.55 g, 197.91 mmol) was added dropwise under ice bath, and stirring was continued for 10 min at this temperature, and then the reaction mixture was transferred to 70°C and stirred for 5 h. After the reaction was complete, the mixture was concentrated under reduced pressure to obtain the title compound 4-2 (9.20 g, 84.21%) as a white solid, which was used directly in the next step.

[0165] Step 2: Synthesis of compound 4-3

[0166] Compound 4-2 (9.20 g, 55.55 mmol) and N-Boc-L-tert-leucine (4.19 g, 18.11 mmol) were dissolved in DCM (30 mL), and DIPEA (15.31 mL, 90.55 mmol) was added, and the mixture was transferred to 0 °C. After that, HATU (10.54 g, 27.16 mmol) and HOAT (3.70 g, 27.16 mmol) were added, and the reaction mixture was transferred to room temperature and stirred for 12 h. After the reaction, hydrochloric acid solution (1M) was added to adjust the pH to 5-6, and then water (50mL) was added to dilute, and extracted with DCM (100mL×3), and the organic phases were combined, washed with saturated brine (100mL×2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 1 / 9) to obtain the title compound 4-3 (4.69g, 75.61%) as a white solid. MS (ESI, pos.ion) m / z: 243.2 [M-Boc+H] + .

[0167] Step 3: Synthesis of compound 4-4

[0168] Compound 4-3 (2.00 g, 5.84 mmol) was dissolved in DCM (2 mL), HCl in 1,4-dioxane (14.71 mL, 58.4 mmol) was added, and the reaction mixture was stirred at room temperature for 1 h. After the reaction was complete, it was concentrated under reduced pressure to give the title compound 4-4 (1.63 g, 100.11%) as a white solid, which was used directly in the next step.

[0169] Step 4: Synthesis of compound 4-5

[0170] Compound 4-4 (1.63 g, 5.85 mmol) and 9-bromo-1-nonene (1.84 g, 8.77 mmol) were dissolved in DMF (15 mL), and potassium carbonate (4.13 g, 29.25 mmol) and sodium iodide (0.88 g, 5.85 mmol) were added. The reaction mixture was heated to 60 °C and stirred. After the reaction was completed, it was cooled to room temperature, diluted with water (30 mL), and then extracted with EA (50 mL × 3). The organic phases were combined, washed with saturated brine (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 1 / 9) to obtain the title compound 4-5 (1.41 g, 65.79%), a yellow oily liquid. MS (ESI, pos.ion) m / z: 367.3 [M+H] + .

[0171] Step 5: Synthesis of Compound 4-6

[0172] Under nitrogen protection, compound 4-5 (0.82 g, 2.24 mmol) was dissolved in DCE (600 mL), heated to 90 ° C and refluxed for 40 min, then a solution of Hoveyda-Grubbs catalyst (0.56 g, 0.57 mmol) in DCE (40 mL) was slowly added dropwise, and the mixture was stirred at the same temperature for 4 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, diluted with hydrochloric acid solution (10 mL, 1N) and water (10 mL), and then washed with EA (30 mL), the pH value of the aqueous phase was adjusted to 7-8, and then extracted with EA (30 mL × 2), the organic phases were combined, washed with saturated brine (20 mL × 2), and dried over anhydrous sodium sulfate. The residue was separated and purified by silica gel column chromatography (EA / PE (V / V) = 2 / 8) to obtain the title compound 4-6 (190 mg, 25.09%), a light yellow oily liquid. MS (ESI, pos.ion) m / z: 339.3 [M+H] + .

[0173] Step 6: Synthesis of Compounds 4-7

[0174] Compound 4-6 (190 mg, 0.56 mmol) was dissolved in a mixture of methanol (3 mL), THF (3 mL) and water (0.5 mL), and then lithium hydroxide monohydrate (75 mg, 1.79 mmol) was added. The reaction mixture was transferred to 60°C and stirred for 2 h. After the reaction was completed, it was concentrated under reduced pressure. The residue was dissolved in a mixed solvent of methanol and dichloromethane (20 mL, MeOH / DCM (V / V) = 1 / 9), and then adjusted to pH 6-7 with a cation exchange resin (pH = 5-6), filtered, and concentrated under reduced pressure to obtain the title compound 4-7 (164 mg, 90.05%) as a yellow solid. MS (ESI, pos.ion) m / z: 325.3 [M+H] + .

[0175] Step 7: Synthesis of Compounds 4-8

[0176] Compound 4-7 (160 mg, 0.49 mmol) and compound 3-1 (140 mg, 0.67 mmol) were dissolved in DMF (5 mL), HOPO (10 mg, 0.088 mmol) was added, and DIPEA (0.57 mL, 3.35 mmol) and EDCI (145 mg, 0.74 mmol) were added at 0°C. The reaction mixture was transferred to room temperature and stirred for 12 h. After the reaction was completed, water (20 mL) was added, and then extracted with EA (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (MeOH / DCM (V / V) = 7 / 93) to obtain the title compound 4-8 (124 mg, 52.65%) as a yellow solid. MS (ESI, pos.ion) m / z: 478.3 [M+H] + .

[0177] Step 8: Synthesis of compound (4)

[0178] Compound 4-8 (124 mg, 0.26 mmol) was dissolved in THF (5 mL), and (1E)-1-methoxy-N-(triethylammonium)sulfonylformimide boronate (160 mg, 0.65 mmol) was added. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, saturated sodium bicarbonate aqueous solution (10 mL) was added to quench the reaction, and then water (10 mL) was added. The reaction mixture was extracted with EA (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (MeOH / DCM (V / V) = 1 / 99) to obtain the title compound (4) (35 mg, 29.33%) as a white solid. MS (ESI, pos.ion) m / z: 460.4 [M+H] + . 1 H NMR (400MHz, CD3OD) δ5.59(dd,J=14.7,7.3Hz,1H),5.47–5.35(m,1H),5.00(dd,J=10.4,5.4Hz,1H),4.32(dd,J=10.3,4.1Hz,1H),3.28 –3.19(m,1H),2.93(s,1H),2.65–2.25(m,8H),2.17–1.99(m,3H),1.96–1.72(m,3H),1.65–1.51(m,2H),1.51–1.34(m,7H),0.96(s,9H).

[0179] Biological tests

[0180] 1. In vitro biochemical reaction evaluation of the inhibitory activity of the compounds of the present invention on SARS-COV-2Mpro / 3CLpro protease

[0181] The fluorescence resonance energy transfer (FRET) method is used, and the detection principle is as follows: Edans is a fluorescent donor, Dabcyl is a fluorescent acceptor or quenching group, and the absorption spectra of these two fluorescent groups overlap to a certain extent. When the distance between the two fluorescent groups is appropriate, the fluorescent energy is transferred from the donor to the acceptor, resulting in the attenuation of the fluorescence intensity of the donor fluorescent molecule itself. Edans and Dabcyl are connected to the two ends of the natural substrate of SARS-COV-2Mpro / 3CLpro protease, namely Dabcyl-KTSAVLQSGFRKME-Edans. When the SARS-COV-2Mpro / 3CLpro protease does not cleave the substrate, the two groups are close enough to cause fluorescence resonance energy transfer, that is, Dabcyl can quench the fluorescence of Edans and cause the fluorescence to be undetectable; when the substrate is cleaved by the SARS-COV-2Mpro / 3CLpro protease, the head and tail ends of the polypeptide are separated, the two groups are separated, and the fluorescence of Edans is no longer quenched by Dabcyl, so the fluorescence of Edans can be detected. In this way, the enzymatic activity of SARS-COV-2Mpro / 3CLpro protease can be detected very sensitively through fluorescence detection. If an inhibitor of SARS-COV-2Mpro / 3CLpro is added to the reaction system, the generation of fluorescence will be inhibited, and the fluorescence intensity is inversely proportional to the inhibitory effect of the inhibitor, so the inhibitory effect of the SARS-COV-2Mpro / 3CLpro protease inhibitor can be detected. The maximum excitation wavelength of Edans is 340nm, and the maximum emission wavelength is 490nm.

[0182] Compound preparation: Dissolve the compound in DMSO to prepare a 20 mM stock solution, further dilute with DMSO to a 200 μM working solution, and then perform 4-fold dilution to 8 concentration points.

[0183] Enzyme activity assay: Use the modified Bio-Tech "New Coronavirus Mpro / 3CLpro Inhibitor Screening Kit" to assay compound activity. Thaw the Assay Buffer and Substrate in the kit and equilibrate to room temperature before use. Place the SARS-COV-2Mpro / 3CLpro protease on ice. Add 92.75μL of Assay Buffer and 0.25μL of SARS-COV-2Mpro / 3CLpro protease to each well. Calculate the required total amount of Assay Buffer and SARS-COV-2Mpro / 3CLpro protease according to the number of samples (including relevant controls). After fully mixing, add 93μL to each well of the 96-well plate. For the blank control, add 93μL of Assay Buffer to each well without adding enzyme. For the 100% enzyme activity control, add 92.75μL of Assay Buffer and 0.25μL of SARS-COV-2Mpro / 3CLpro protease to each well of the sample well. Then add 5 μL of the above serially diluted compound working solution to each sample well, set up 2 replicates for each concentration, and use Shionogi 3CLpro inhibitor S-217622 as the positive control compound. Add 5 μL DMSO to each well of the blank control and 100% enzyme activity control. Incubate the 96-well plate at 37°C for 10 minutes. Quickly add 2 μL of matrix to each well and mix well. The total reaction system is 100 μL, and the highest final concentration of the compound tested is 10 μM. After incubation at 37°C in the dark for 25 minutes, use a multifunctional enzyme label to measure the fluorescence intensity. The excitation wavelength is 340 nm and the emission wavelength is 490 nm. Calculate the inhibition percentage of each sample. The calculation formula is as follows:

[0184] Inhibition rate (%) = (RFU 100% enzyme activity control - RFU sample) / (RFU 100% enzyme activity control - RFU blank control) × 100%

[0185] RFU, Relative Fluorescence Unit (relative fluorescence value).

[0186] Graphpad Prism 7 software was used to process the concentration-inhibition rate, and the IC of the compound inhibiting SARS-COV-2Mpro / 3CLpro protease was calculated by a four-parameter nonlinear regression model. 50 The compounds of the present invention have good inhibitory activity against SARS-COV-2Mpro / 3CLpro protease. The experimental results of some of the compounds of the present invention are shown in Table A.

[0187] Table A: Experimental results of the inhibitory activity of some compounds of the present invention on SARS-COV-2Mpro / 3CLpro protease

[0188] Test compound <![CDATA[IC 50 (μM) <!-- 23 -->]]> Compound (1) 0.01 Compound (2) 0.50 Compound (3) 0.18 Compound (4) 0.38

[0189] Conclusion: The experimental results show that the compounds of the present invention have good inhibitory activity against SARS-COV-2Mpro / 3CLpro protease.

[0190] 2. Anti-SARS-CoV-2 activity detection of samples

[0191] Methods and steps: Vero E6 was inoculated in a 96-well cell culture plate at 1.0×104 cells per well and cultured overnight at 37°C for later use. After adding the gradient diluted drugs, SARS-CoV-2 was added to infect the cells, and the MOI was about 0.01. After culturing in a 37°C cell culture incubator for 72 hours, the cytopathic effect (CPE) was observed under a microscope, 100 μL of the culture supernatant was removed, 100 μL of CellTiter-GloTM reagent was added, and the chemiluminescent signal was detected after mixing. The experiment set up blank control wells (normal cells), virus control wells (no drug was added after virus infection), and positive drug control wells (remdesivir was added after infection).

[0192] Inhibition rate (%) = (100-sample well) / (blank control-virus control) × 100

[0193] The calculated cell viability and inhibition rate were plotted using Graphpad Prism 8. 50 The calculation formula of (cell half pathological concentration) is: Nonlin fit / Sigmoidal dose-response (variable slope). The experimental results show that the compounds of the present invention have good inhibitory activity against SARS-CoV-2.

[0194] 2. Toxicity test of Vero E6 cells of samples

[0195] The experiment used the CellTiter-GloTM (Promega) kit to detect the cytotoxic effects of samples.

[0196] Experimental principle: The CellTiter-GloTM kit detects the number of live cells in the culture by quantitatively measuring ATP. The respiration and other life activities of metabolically active cells can produce ATP. The kit uses a stable glow signal generated by luciferase, and luciferase requires the participation of ATP in the luminescence process. Add CellTiter-GloTM reagent to the cell culture medium and measure the luminescence value. The light signal is proportional to the amount of ATP in the system, and the amount of ATP is positively correlated with the number of live cells. Therefore, the light signal value can reflect the number of live cells.

[0197] Experimental steps: Vero E6 was inoculated in a 96-well cell culture plate at 1.0×104 cells per well and cultured overnight at 37°C. After adding the gradient diluted drugs, the cell plate was cultured in a 37°C cell culture incubator for 72 hours, 100 μl of the culture supernatant was removed, 100 μl of CellTiter-GloTM reagent was added, and the chemiluminescent signal was detected after mixing.

[0198] CC 50 The calculation formula of (cell half pathological concentration) is: Nonlin fit / Sigmoidal dose-response (variable slope). Conclusion: The experimental results show that the compounds of the present invention have low toxicity to cells.

[0199] 2. Liver microsomal stability test of the compounds of the present invention in different species

[0200] Experimental method: Add 30 μL of the mixed solution of the test compound and liver microsomes to a 96-well plate and make two samples in parallel. After pre-incubation at 37°C for 10 minutes, add 15 μL of NADPH solution (6 mM) at the time point. The final concentration of the test compound is 1 μM, the concentration of liver microsomes is 0.5 mg / mL, and the final concentration of NADPH is 2 mM. Incubate for 0, 15, 30, and 60 minutes respectively. After the incubation, add 150 mL of acetonitrile (containing internal standard) to the mixed system. The sample diluted with acetonitrile is centrifuged at 4000 rpm for 5 minutes, and 150 μL of supernatant is taken for LC-MS / MS analysis. The compounds of the present invention have good stability in liver microsomes of different species. Conclusion: The compounds of the present invention have good stability in liver microsomes of different species.

[0201] 3. Test of the tissue distribution of the compounds of the present invention in mice.

[0202] Experimental method: 12 male C57BL / 6 mice were gavaged with 10 mg / kg of the drug. The plasma, whole blood and tissues (heart, liver, spleen, lung, kidney, brain, stomach, small intestine, skin, muscle, fat, testis, epididymis) of the animals were collected at 0.5h, 2h, 7h and 24h after administration. Three animals were collected at each time point. After tissue collection, the tissues were weighed and homogenized according to the ratio of adding 500μL or 1000μL of methanol to 100mg of tissue, and then centrifuged. After centrifugation, the supernatant was taken to obtain tissue homogenate samples. 20μL of plasma, whole blood and tissue homogenate samples were taken, 150μL of acetonitrile was added to precipitate proteins, vortexed and mixed, and the supernatant was taken by centrifugation. 100μL of supernatant was added with 150μL of ultrapure water for dilution and LC-MS / MS analysis was performed.

[0203] Conclusion: The compounds of the present invention have higher concentrations in the main target organs of male C57BL / 6 mice, such as the lung and heart.

[0204] 4. Test on the inhibitory effect of the compounds of the present invention on human liver microsomal CYP enzymes.

[0205] Experimental purpose: To evaluate the inhibitory effects of test compounds on the main metabolic enzymes CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4 in microsomes using the human liver microsome system.

[0206] Experimental method: The concentration of the test substance in the final incubation system is 10 μM, and it is incubated with a suspension of human liver microsomes containing cytochrome P450 enzymes CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6 and CYP3A4. It is known that the substrate is metabolized and converted by the corresponding single enzyme to generate specific metabolites, and the metabolites are determined by the UPLC-MS / MS method. The relative inhibition rate (%) is calculated by the percentage reduction of metabolite generation in the sample group and the solvent control group. If the relative inhibition rate (%) is greater than 40%, the test substance may have the possibility of inhibiting the corresponding metabolic enzyme. The experimental results show that the compounds of the present invention have basically no inhibitory effect on the main human liver microsomal CYP enzymes.

[0207] 5. Pharmacokinetics of the compounds of the present invention in beagle dogs, mice and rats

[0208] (1) Beagle dog PK test

[0209] PK determination experimental method of the compound of the present invention in beagle dogs (purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., weight 10-12 kg, male, age 10-12 months, oral administration 3 per group, intravenous injection 3 per group)

[0210] Beagle dogs were given 2.5 mg / kg or 5 mg / kg of the test compound by oral gavage or 0.5 mg / kg or 1 mg / kg or 2 mg / kg by intravenous injection. After administration, venous blood was collected at time points (0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours) and collected in anticoagulant tubes with EDTA-K2. After liquid-liquid extraction, the plasma samples were quantitatively analyzed by multiple reaction ion monitoring (MRM) on a triple quadrupole tandem mass spectrometer. The pharmacokinetic parameters were calculated using the non-compartmental model method using WinNonlin 6.3 software. The experimental results show that the compounds of the present invention have good pharmacokinetic properties in beagle dogs.

[0211] (2) Mouse PK test experiment

[0212] PK determination of the compound of the present invention in ICR mice (purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., body weight 20-25g, male, age 45-60 days, oral administration 3 mice per group, intravenous injection 3 mice per group) Experimental method: ICR mice were given 10 mg / kg of the test compound by oral gavage or 2 mg / kg or 10 mg / kg by tail vein injection. Blood was collected from the orbital vein at time points (0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours) after administration and collected in anticoagulation tubes with EDTA-K2. After liquid-liquid extraction, the plasma samples were quantitatively analyzed on a triple quadrupole tandem mass spectrometer in a multiple reaction ion monitoring (MRM) mode. The pharmacokinetic parameters were calculated using the non-compartmental model method using WinNonlin 6.3 software. The experimental results show that the compound of the present invention has good pharmacokinetic properties in ICR mice.

[0213] (3) PK test experiment in SD rats

[0214] The PK determination experimental method of the compound of the present invention in SD rats (purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., body weight 200-250g, male, age 2-3 months, oral administration 3 rats per group, intravenous injection 3 rats per group) is as follows: the rats are given 2.5mg / kg or 5mg / kg of the test compound by oral gavage or 1mg / kg by intravenous injection. After administration, venous blood is collected at time points (0.083, 0.25, 0.5, 1, 2, 5, 7 and 24 hours) and collected in anticoagulation tubes with EDTA-K2. After liquid-liquid extraction, the plasma samples are quantitatively analyzed by multiple reaction ion monitoring (MRM) on a triple quadrupole tandem mass spectrometer. The pharmacokinetic parameters are calculated by the non-compartmental model method using WinNonlin 6.3 software. The pharmacokinetic experimental data show that the exposure of the compound of the present invention in SD rats is large, indicating that the compound of the present invention is well absorbed in SD rats and has good bioavailability.

[0215] (4) PK test experiment in cynomolgus monkey

[0216] PK determination of the compound of the present invention in cynomolgus monkeys (purchased from Guangdong Chunsheng Biotechnology Development Co., Ltd., body weight 3-6kg, male, age 4-6 years old, 3 per group for oral administration, 3 per group for intravenous injection) Experimental method: 2.5 mg / kg or 5 mg / kg of the test compound was administered orally to cynomolgus monkeys or 0.5 mg / kg or 1 mg / kg was injected intravenously. After administration, venous blood was collected at time points (0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours) and collected in anticoagulant tubes with EDTA-K2. After liquid-liquid extraction, the plasma samples were quantitatively analyzed by multiple reaction ion monitoring (MRM) on a triple quadrupole tandem mass spectrometer. The pharmacokinetic parameters were calculated by the non-compartmental model method using WinNonlin 6.3 software. The pharmacokinetic experimental data showed that the compound of the present invention had good pharmacokinetic properties in cynomolgus monkeys.

[0217] Although the present invention has been described in detail above by means of general description, specific implementation methods and tests, it is obvious to those skilled in the art that some modifications or improvements may be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A compound, which is a compound as represented by formula (I) or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof of the compound as represented by formula (I), in, L is C 2-8 Alkylene or C 2-12 Alkenylene, wherein the C 2-12 Alkylene and C 2-8 Each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 groups independently selected from deuterium, F, Cl, Br and C 1-4 substituted by an alkyl substituent; Each R 1 , R 2 , R 3 , R 4 H, deuterium, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Haloalkyl; R 5 H, deuterium, F, Cl, Br, -CN, C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 2-6 Alkenyl, C 1-6 Haloalkyl or C 3-6 Cycloalkyl; R 6 C 1-6 Alkyl, C 2-6 Alkenyl or C 1-6 Haloalkyl; R is H, deuterium, F, Cl, Br, -CN, -NO2, -COOH, -C 1-6 Alkylene -COOH, amino, -C(=O)OC 1-6 Alkyl, -C 1-6 Alkylene-C(=O)OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 6-14 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6 Cycloalkyl, heterocyclic group consisting of 3-10 ring atoms, bicyclic heteroaryl consisting of 9-10 ring atoms, -C 2-6 Alkenylene-C(=O)OR 6 or -C(=O)CH2-OC(=O)R 7 , wherein the -C 1-6 Alkylene -COOH, amino, -C(=O)OC 1-6 Alkyl, -C 1-6 Alkylene-C(=O)OC 1-6 Alkyl, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Hydroxyalkyl, C 6-14 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6 Cycloalkyl, heterocyclic group consisting of 3-10 ring atoms, bicyclic heterocyclic group consisting of 9-10 ring atoms, -C 2-6 Alkenylene-C(=O)OR 6 C 2-6 Alkenylene and -C(=O)CH2-OC(=O)R 7 The CH2 in each of the above is independently unsubstituted or substituted by 1, 2 or 3 independently selected from deuterium, F, Cl, Br, -OH, CN, C 1-6 Alkyl, C 1-6 Haloalkyl, -COOH, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy and C 1-6 substituted with a hydroxyalkoxy substituent; R 7 C 6-14 aryl, a monocyclic heteroaryl consisting of 5-6 ring atoms, or a bicyclic heteroaryl consisting of 9-10 ring atoms, wherein the C 6-14 The aryl group, the monocyclic heteroaryl group consisting of 5-6 ring atoms and the bicyclic heteroaryl group consisting of 9-10 ring atoms are each independently unsubstituted or substituted with 1, 2, 3 or 4 R w1 replaced by; Each R w1 are independently deuterium, F, Cl, Br, -CN, -NO2, -COOH, amino, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 Halogenated alkoxy.

2. The compound according to claim 1, wherein L is C 4-12 Alkylene or C 4-12 Alkenylene, wherein the C 4-12 Alkylene and C 4-12 Each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 groups independently selected from deuterium, F, Cl, Br and C 1-3 substituted by an alkyl substituent; or L is C 6-12 Alkylene or C 6-12 Alkenylene, wherein the C 6-12 Alkylene and C 6-12 each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl; or L is C 8-11 Alkylene or C 8-11 Alkenylene, wherein the C 8-11 Alkylene and C 8-11 each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl; or L is C 9-10 Alkylene or C 9-10 Alkenylene, wherein the C 9-10 Alkylene and C 9-10 each alkenylene group is independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl; or L is -(CH2)9-, -(CH2) 10 -, -(CH2)6-CH=CHCH2-, -(CH2)7-CH=CHCH2-, -(CH2)5-CH=CHCH2CH2- or -(CH2)- CH2CH=CHCH2-, wherein -(CH2)9-, -(CH2) 10 -, -(CH2)6-CH=CHCH2-, -(CH2)7-CH=CHCH2-, -(CH2)5-CH=CHCH2CH2- and -(CH2)-CH2CH=CHCH2- are each independently unsubstituted or substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, methyl, ethyl, n-propyl and isopropyl.

3. The compound according to claim 1 or 2, wherein Each R 1 , R 2 , R 3 and R 4 H, deuterium, C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 2-4 Alkenyl or C 1-4 haloalkyl; or Each R 1 , R 2 , R 3 and R 4 are independently H, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CD3, CHD2, -CH2CD3, vinyl, propenyl, allyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, or -CH2CH2CF3; R 5 H, deuterium, F, Cl, Br, -CN, C 1-4 Alkyl, deuterated C 1-4 Alkyl, C 2-4 Alkenyl, C 1-4 haloalkyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; or R 5 is H, F, Cl, Br, -CN, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CD3, CHD2, -CH2CD3, vinyl, propenyl, allyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; R 6 C 1-4 Alkyl, C 2-4 Alkenyl or C 1-4 haloalkyl; or R 6 It is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, vinyl, propenyl, allyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2 or -CH2CH2CF3.

4. The compound according to any one of claims 1 to 3, wherein R is H, deuterium, F, Cl, Br, -CN, -NO2, -COOH, -C 1-4 Alkylene -COOH, amino, -C(=O)OC 1-4 Alkyl, -C 1-4 Alkylene-C(=O)OC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 6-12 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6 Cycloalkyl, heterocyclic group consisting of 3-6 ring atoms, bicyclic heteroaryl consisting of 9-10 ring atoms, -C 2-4 Alkenylene-C(=O)OR 6 or -C(=O)CH2-OC(=O)R 7 , wherein the -C 1-4 Alkylene -COOH, amino, -C(=O)OC 1-4 Alkyl, -C 1-4 Alkylene-C(=O)OC 1-4 Alkyl, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy, C 1-4 Hydroxyalkyl, C 6-12 Aryl, monocyclic heteroaryl consisting of 5-6 ring atoms, C 3-6 Cycloalkyl, heterocyclic group consisting of 3-6 ring atoms, bicyclic heteroaryl consisting of 9-10 ring atoms, -C 2-4 Alkenylene-C(=O)OR 6 C 2-4 Alkenylene and -C(=O)CH2-OC(=O)R 7 The CH2 in each of the above is independently unsubstituted or substituted by 1, 2 or 3 independently selected from deuterium, F, Cl, Br, -OH, CN, C 1-4 Alkyl, C 1-4 Haloalkyl, -COOH, C 1-4 Hydroxyalkyl, C 1-4 Alkoxy and C 1-4 The hydroxyl group is substituted with an alkoxy group.

5. The compound according to any one of claims 1 to 4, wherein R is H, deuterium, F, Cl, Br, -CN, -NO2, -COOH, -C 1-2 Alkylene -COOH, amino, -C(=O)OC 1-3 Alkyl, -C 1-2 Alkylene-C(=O)OC 1-3 Alkyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-l-propoxy, 2-butoxy, C 1-3 Halogenated alkoxy, C 1-3 hydroxyalkyl, phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, propyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, thietanyl, oxirane, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, dihydropyridine, morpholinyl, thiomorpholinyl, piperazinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothiophenyl, indolyl, purinyl, quinolinyl, isoquinolinyl, -CH=CH-C(=O)OR 6 、-CH=CHCH2-C(=O)OR 6 、-CH2CH=CH-C(=O)OR 6 、-CH2CH=CHCH2-C(=O)OR 6 、-CH=CHCH2CH2-C(=O)OR 6 、-CH2CH2CH=CH-C(=O)OR 6 or -C(=O)CH2-OC(=O)R 7 , wherein the -C 1-2 Alkylene -COOH, amino, -C(=O)OC 1-3 Alkyl, -C 1-2 Alkylene-C(=O)OC 1-3 Alkyl, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, C 1-3 Halogenated alkoxy, C 1-3 hydroxyalkyl, phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, propyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, oxetanyl, thietanyl, oxirane, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, dihydropyridine, morpholinyl, thiomorpholinyl, piperazinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothiophenyl, indolyl, purinyl, quinolinyl and isoquinolinyl, -CH=CH-C(=O)OR 6 CH=CH、-CH=CHCH2-C(=O)OR 6 CH=CHCH2、-CH2CH=CH-C(=O)OR 6 CH2CH=CH、-CH2CH=CHCH2-C(=O)OR 6 CH2CH=CHCH2、-CH=CHCH2CH2-C(=O)OR 6 CH=CHCH2CH2、-CH2CH2CH=CH-C(=O)OR 6 CH2CH2CH=CH and -C(=O)CH2-OC(=O)R 7 wherein CH2 is independently unsubstituted or replaced by 1, 2 or 3 independently selected from deuterium, F, Cl, Br, -OH, CN, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, -COOH, C 1-3 Hydroxyalkyl, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy and C 1-3 The hydroxyl group is substituted with an alkoxy group.

6. The compound according to any one of claims 1 to 5, wherein R 7 is phenyl, naphthyl, a monocyclic heteroaryl consisting of 5 ring atoms, a monocyclic heteroaryl consisting of 6 ring atoms, a bicyclic heteroaryl consisting of 9 ring atoms, or a bicyclic heteroaryl consisting of 10 ring atoms, wherein The phenyl, naphthyl, monocyclic heteroaryl composed of 5 ring atoms, monocyclic heteroaryl composed of 6 ring atoms, bicyclic heteroaryl composed of 9 ring atoms and bicyclic heteroaryl composed of 10 ring atoms are each independently unsubstituted or substituted with 1, 2, 3 or 4 R w1 replaced by; or R 7 is phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothienyl, indolyl, purinyl, quinolyl or isoquinolyl, wherein the phenyl, naphthyl, furanyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, thienyl, pyridinyl, 1,3,5-triazinyl, pyrazinyl, pyridazinyl, pyrimidinyl, benzopyrazolyl, benzopyrrolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzothienyl, indolyl, purinyl, quinolyl and isoquinolyl are each independently unsubstituted or substituted with 1, 2, 3 or 4 R w1 replaced by; Each R w1 are independently deuterium, F, Cl, Br, -CN, -NO2, -COOH, amino, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy or C 1-4 haloalkoxy; or Each R w1 are independently deuterium, F, Cl, Br, -CN, -NO2, -COOH, amino, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, -CH2F, -CH2Cl, -CHF2, -CHCl2, -CF3, -CH2CH2F, -CH2CH2Cl, -CH2CHF2, -CH2CHCl2, -CHFCH2F, -CHClCH2Cl, -CH2CF3, -CH(CF3)2, -CF2CH2CH3, -CH2CH2CH2F, -CH2CH2CHF2, -CH2CH2CF3, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy or C 1-3 Halogenated alkoxy.

7. A compound according to any one of claims 1 to 6, which is a compound as shown in formula (II) or a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or prodrug thereof of a compound as shown in formula (II), in, Each --- is independently a single bond or a double bond; n is 1 or 2, and R has the definition as described in any one of claims 1 to 6.

8. The compound according to any one of claims 1 to 7, which is one of the following compounds or its stereoisomers, tautomers, nitrogen oxides, solvates, metabolites, pharmaceutically acceptable salts or prodrugs:

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8, optionally further comprising a pharmaceutically acceptable excipient or any combination thereof; Optionally, the pharmaceutical composition further comprises other antiviral drugs, and the other antiviral drugs are enstevir, remdesivir, favipiravir, namatevir, ribavirin, monoupivir, baricitinib, monoupivir, paxlovid, azithromycin, camostat, nafamostat, ritonavir, lopinavir, nitazoxanide, chloroquine, hydroxychloroquine, darunavir, arbidol, resveratrol, interferon α, interferon β, chlorpromazine, imatinib, oseltamivir, loperamide, darunavir, nelfinavir, Jinhua Qinggan Granules, Shufeng Jiedu Capsules, Lianhua Qingwen Capsules or any combination thereof.

10. Use of the compound described in any one of claims 1 to 8 or the pharmaceutical composition described in claim 9 in the preparation of a medicament for preventing, treating, curing or alleviating a disease or symptom caused by a coronavirus, wherein the coronavirus is HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV or SARS-CoV-2.

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