Application of spiro compound in preparation of 3CLpro protease inhibitor or drug

Abstract

The invention discloses application of spiro compounds in preparation of 3CLpro protease inhibitors or drugs, and belongs to the technical field of medicines. The application refers to application of spiro compounds in preparation of 3CLpro protease inhibitors or drugs, and the drugs are drugs for treating and / or preventing diseases caused by coronavirus infection. The spiro compound has better inhibitory activity on main protease activity and oxidase endurance capacity, has good anti-coronavirus activity, can be used for preventing or treating diseases caused by coronavirus infection, and has a good medicinal prospect.

Description

[0001] This application is a divisional of the Chinese invention patent with application date of December 29, 2023, application number 2023118576791, and name of spirocyclic compounds, preparation methods thereof and uses thereof. Technical Field

[0002] The present invention belongs to the field of medical technology, and specifically relates to an application of a spirocyclic compound in the preparation of a 3CLpro protease inhibitor or a drug, and more specifically discloses an application of a spirocyclic compound in the preparation of a drug for preventing and / or treating a disease associated with 3CLpro protease or a disease caused by coronavirus infection. Background Art

[0003] Although vaccines have been widely used as a means to enhance herd immunity, SARS-CoV-2 itself is very prone to mutation and produces various mutant strains. When new mutant strains begin to spread, people who have been vaccinated against the original strains will still be infected, and the vaccine will lose its protective effect. Therefore, the development of anti-coronavirus drugs remains a top priority.

[0004] The anti-coronavirus drugs currently under development can be divided into several categories according to different mechanisms of action. Among them, the main protease inhibitors such as main protease inhibitors and RNA polymerase inhibitors have attracted widespread attention in the industry due to their respective excellent characteristics. The relevant content of main protease inhibitors is now explained as follows:

[0005] The main protease (also known as 3CLpro, Mpro or 3CL protease) is a three-domain cysteine ​​protease encoded by the virus's own genome, which plays an important role in the replication of SARS-CoV-2. The SARS-CoV-2 genome encodes two polyproteins (pp1a and pp1ab) and four structural proteins. These polyproteins are cleaved by the key SARS-CoV-2 main protease at 11 different sites to produce shorter non-structural proteins that are essential for viral replication. These non-structural proteins include RdRp, helicase and the main protease itself, which are responsible for various stages of SARS-CoV-2 viral replication. In the main protease, the structure that plays the main catalytic role is the Cys145-His41 catalytic dyad located in the cleft between its domain I and domain II, and this dyad and the substrate recognition pocket are highly conserved in the main protease of different subtypes of coronaviruses. Because the main protease plays a vital role in the viral replication process and its active structure is relatively conserved, it can become a key drug target against coronaviruses. In addition, there is no protein similar to the main protease in the human body, so the safety of drugs that can inhibit the main protease can also be guaranteed. Therefore, the development of corresponding main protease inhibitor drugs targeting the main protease has a good prospect.

[0006] The main protease usually has Cys145 and four active sites. Existing main protease inhibitors are designed by designing the target head to bind to Cys145, and according to the characteristics of the four active sites, the corresponding molecular structure is designed to achieve specific binding with the above sites, thereby inhibiting the activity of the main protease and preventing SARS-CoV-2 from replicating in the body.

[0007] Patent WO2021250648 and Pfizer's published literature disclose a main protease inhibitor: Among them, the representative specific compound PF-07321332 (Nimategravir) The drug has good anti-SARS-CoV-2 activity, and has certain oral availability and safety, and can be used as an oral drug to treat COVID-19. However, when the drug is used orally, it still needs to be used in combination with ritonavir, and its tolerance to oxidase is still not high.

[0008] The exemplary specific compounds disclosed in patent WO2023009187 are shown in the following formula. Such compounds have poor oxidative stability.

[0009] Summary of the invention

[0010] In order to overcome the defects of the prior art, such as the small number of compounds that inhibit the main protease and poor oxidase tolerance, the present invention provides a spirocyclic compound for use in the preparation of 3CLpro protease inhibitors or drugs. Specifically, the use of spirocyclic compounds or pharmaceutically acceptable salts thereof in the preparation of 3CLpro protease inhibitors or drugs, such compounds have comparable or better main protease activity inhibition ability and better oxidase tolerance than namatevir, achieve good antiviral activity and human liver microsome stability, and are expected to be clinically unnecessary to be used in conjunction with ritonavir drugs, have a longer oxidative metabolism time in the body, reduce the number of oral administrations per day, and help improve drug compliance.

[0011] The present invention provides the following technical solutions to solve the above technical problems.

[0012] The present invention provides a compound as shown in formula I or a pharmaceutically acceptable salt thereof,

[0013]

[0014] in,

[0015] X 1 is N or CH; X 2 For-NR X2 , CR X3 or CR X4 R X5 ;

[0016] R X2 , R X3 , R X4 and R X5 are independently -H, -C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl;

[0017] R 3 -H, oxo, halogen, C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl;

[0018] is a single bond or a double bond;

[0019] R 1 and R 2 are independently H, -C(O)R 11 、-C(O)OR 12 、-S(O)2R 13 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the “5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S” and the “3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S” are independently optionally replaced by 1, 2 or 3 R a replace;

[0020] R 11 , R 12 and R 13 Independently -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S", the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the “5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S” and the “3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S” are independently optionally replaced by 1, 2 or 3 R b replace;

[0021] R 4 and R 5 are independently H, -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R c replace;

[0022] R a are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl;

[0023] R b are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl;

[0024] R c are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl;

[0025] R 6 is cyano; R 7 is hydrogen; R 8 is a halogen; n is 0 or 1;

[0026] When X 1 CH, X 2 CR X3 or CR X4 R X5 , R 1 and R 2 One of the groups is H and the other is -C(O)R 11 、-C(O)OR 12 or -S(O)2R 13 , R 11 , R 12 and R 13 Independently -C 1-10 When the -C 1-10 The alkyl group is replaced by 1, 2 or 3 R a replace.

[0027] In some embodiments, in the compound as shown in Formula I or a pharmaceutically acceptable salt thereof, certain groups have the following definitions, and the definitions of the unmentioned groups are as described in any embodiment of the present invention (hereinafter referred to as "a preferred embodiment").

[0028] In a preferred embodiment, R X2 and R X3 In the -C 1-6 The alkyl group may independently be -C 1-4 An alkyl group, such as a methyl group.

[0029] In a preferred embodiment, R X2and R X3 wherein the -C substituted by 1, 2 or 3 halogens 1-6 -C in the alkyl group 1-6 The alkyl group may independently be -C 1-4 An alkyl group, such as a methyl group.

[0030] In a preferred embodiment, R 1 , R 2 , R 11 , R 12 , R 13 , R 4 , R 5 , R a , R b and R c In the -C 1-10 Alkyl and the -C substituted by 1, 2 or 3 halogens 1-6 -C in the alkyl group 1-6 The alkyl group may independently be -C 1-6 Alkyl, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl.

[0031] In a preferred embodiment, R 1 , R 2 , R 11 , R 12 and R 13 In the -C 3-10 Cycloalkyl groups may independently be -C 3-6 Cycloalkyl, for example cyclopropyl or cyclobutyl.

[0032] In a preferred embodiment, R a and R c In the -C 3-6 Cycloalkyl groups may independently be cyclopropyl or cyclobutyl.

[0033] In a preferred embodiment, R 1 , R 2 , R 11 , R 12 and R 13 In the -C 6-10 Aryl groups may independently be phenyl or naphthyl.

[0034] In a preferred embodiment, R 1 and R 2 In the above, the “heteroatom selected from 1, 2 or 3 of N, O and S, and the number of heteroatoms is 1, 2 or 3, 5-10 membered heteroaryl group” can be independently “heteroatom selected from N, and the number of heteroatoms is 1 or 2, 5-1 membered heteroaryl group”, preferably “heteroatom selected from N, and the number of heteroatoms is 1 or 2, 5-6 membered heteroaryl group”, more preferably pyridyl (for example, ) or diazolyl (e.g., 1H-pyrazolyl, for example, ).

[0035] In a preferred embodiment, R X2 , R X3 , R X4 , R X5 , R a , R b and R c In the above, the halogen may be F.

[0036] In a preferred embodiment, the X 1 is N or CH.

[0037] In a preferred embodiment, the X 2 NR X2 or CR X3 .

[0038] In a preferred embodiment, the R X2 For -C 1-6 alkyl.

[0039] In a preferred embodiment, the R X3 is H or -C substituted by 1, 2 or 3 halogens 1-6 alkyl.

[0040] In a preferred embodiment, the R 3 It is -H or oxo.

[0041] In a preferred embodiment, the R 1 -C(O)R 11 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 Aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group and the “5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S” are independently optionally replaced by 1, 2 or 3 R a replace.

[0042] Preferably, the R 1 -C(O)R 11 , -C 1-4 Alkyl, -C 3-6 Cycloalkyl, phenyl or "a 5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen"; the -C1-4 Alkyl, the -C 3-6 The cycloalkyl group, the phenyl group and the “5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen” are independently optionally substituted by 1, 2 or 3 R a replace.

[0043] More preferably, the R 1 -C(O)R 11 , phenyl or "a 5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen"; the phenyl group and the "a 5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen" are independently optionally replaced by 1, 2 or 3 R a replace.

[0044] In a preferred embodiment, the R 1 Can

[0045] Best More preferred

[0046] In a preferred embodiment, the R 2 For H.

[0047] In a preferred embodiment, the R 11 For -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R b replace.

[0048] Preferably, the R 11 For -C 1-4 Alkyl, -C 3-6 Cycloalkyl or phenyl; the -C 1-4 Alkyl, the -C 3-6 The cycloalkyl and the phenyl are optionally substituted with 1, 2 or 3 halogens;

[0049] More preferably, the R 11 For -C 3-6 Cycloalkyl.

[0050] In a preferred embodiment, the R 11 It is -CF3, cyclopropyl or phenyl; preferably cyclopropyl.

[0051] In a preferred embodiment, the R 4For -C 1-10 Alkyl; the -C 1-10 The alkyl group is optionally substituted by 1, 2 or 3 R c replace.

[0052] Preferably, the R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted with 1 or 2 R c substituted, said Rc is F, cyclopropyl, cyclobutyl or methyl; more preferably, said R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted with 1 cyclopropyl group.

[0053] In a preferred embodiment, the R 4 for Preferably More preferred

[0054] In a preferred embodiment, the R 5 For H.

[0055] In a preferred embodiment, the R a are independently cyano, halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl.

[0056] In a preferred embodiment, the R a It may independently be cyano, halogen, methyl or cyclopropyl; preferably cyano, F or methyl.

[0057] In a preferred embodiment, the R b is independently halogen, preferably F.

[0058] In a preferred embodiment, the R c are independently halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl.

[0059] In a preferred embodiment, the R c It can be halogen, methyl, cyclopropyl or cyclobutyl; preferably cyclopropyl.

[0060] In a preferred embodiment, n is 0.

[0061] In a preferred embodiment, the Can Best

[0062] In a preferred embodiment, the Can Best More preferred

[0063] In a preferred embodiment, the compound represented by formula I may be a compound represented by formula IA,

[0064]

[0065] Wherein, * represents a chiral carbon atom of S configuration or R configuration;

[0066] Preferably, Formula IA is Formula IA-1 or Formula IA-2:

[0067]

[0068] In a preferred embodiment, the compound as shown in formula I may be a compound as shown in formula I-1, a compound as shown in formula I-2 or a compound as shown in formula I-3, preferably a compound as shown in formula I-2 or a compound as shown in formula I-3; more preferably a compound as shown in formula I-3:

[0069]

[0070] In a preferred embodiment, the compound represented by formula I-1 may be a compound represented by formula I-1-A and / or a compound represented by formula I-1-B.

[0071]

[0072] In a preferred embodiment, the compound represented by formula I-2 may be a compound represented by formula I-2-A and / or a compound represented by formula I-2-B.

[0073]

[0074] In a preferred embodiment, the compound represented by formula I-3 may be a compound represented by formula I-3-A and / or a compound represented by formula I-3-BA.

[0075]

[0076] In a preferred embodiment, each substituent in the compound of Formula I is Scheme 1, Scheme 2, Scheme 3, Scheme 4, Scheme 5 or Scheme 6:

[0077] Scenario 1:

[0078] The X 1 is N;

[0079] The X 2 NR X2 , CRX3 or CR X4 R X5 ;

[0080] The R X2 , the R X3 , the R X4 and the R X5 are independently -H, -C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl;

[0081] The R 3 is H or oxo;

[0082] Said is a single bond or a double bond;

[0083] The R 1 and the R 2 are independently H, -C(O)R 11 、-C(O)OR 12 、-S(O)2R 13 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S", the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the “heteroatoms selected from 1, 2 or 3 of N, O and S, the number of heteroatoms is 1, 2 or 3 5-10 membered heteroaryl group” and the “heteroatoms selected from 1, 2 or 3 of N, O and S, the number of heteroatoms is 1, 2 or 3 3-10 membered heterocycloalkyl group” are independently optionally replaced by 1, 2 or 3 R a replace;

[0084] The R 11 , the R 12 and the R 13 Independently -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the “5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S” and the “3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S” are independently optionally replaced by 1, 2 or 3 R b replace;

[0085] The R 4 and the R 5 are independently H, -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R c replace;

[0086] The R a are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl;

[0087] The R b are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl;

[0088] The R c are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl;

[0089] The R 6 is cyano; said R 7 is H; said R 8 is halogen; said n is 0 or 1;

[0090] Scenario 2:

[0091] The X 1 for CH;

[0092] The X 2 NR X2 , CR X3 or CR X4 R X5 ;

[0093] The R X2 , the R X3 , the R X4 and the R X5 are independently -H, -C 1-6 Alkyl, or -C substituted by 1, 2 or 3 halogens 1-6 alkyl;

[0094] The R 3 is -H or oxo;

[0095] Said is a single bond or a double bond;

[0096] The R 1 and the R 2 are independently H, -C(O)R 11 or "a 5-10 membered heteroaryl group wherein the heteroatom is selected from 1, 2 or 3 of N, O and S, and the number of the heteroatom is 1, 2 or 3";

[0097] The R 11 For 1, 2 or 3 R b Replaced-C 1-10 alkyl;

[0098] The R 4 and the R 5 are independently H, -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R c replace;

[0099] The R b are independently halogen;

[0100] The R c are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-10 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl;

[0101] The R6 is cyano; said R 7 is H; said R 8 is halogen; said n is 0 or 1;

[0102] Solution 3:

[0103] The X 1 is N;

[0104] The X 2 NR X2 or CR X3 ;

[0105] The R X2 , the R X3 , the R X4 and the R X5 are independently -H, -C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl;

[0106] The R 3 is -H or oxo;

[0107] Said is a single bond or a double bond;

[0108] The R 1 H, -C(O)R 11 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 Aryl or "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group and the “5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S” are independently optionally replaced by 1, 2 or 3 R a replace;

[0109] The R 2 is H;

[0110] The R 11 For -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R breplace;

[0111] The R 4 For -C 1-10 Alkyl; the -C 1-10 The alkyl group is optionally substituted by 1, 2 or 3 R c replace;

[0112] The R 5 is H;

[0113] The R a are independently cyano, halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl;

[0114] The R b are independently halogen;

[0115] The R c are independently cyano, halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl;

[0116] The R 6 is cyano; said R 7 is H; said R 8 is a halogen; said n is 0;

[0117] Solution 4:

[0118] The X 1 CH, X 2 for CH;

[0119] The R 3 is H;

[0120] Said is a double bond;

[0121] The R 1 -C(O)R 11 or "a 5-10 membered heteroaryl group wherein the heteroatom is selected from 1, 2 or 3 of N, O and S, and the number of the heteroatom is 1, 2 or 3";

[0122] The R 2 is H;

[0123] The R 11 For 1, 2 or 3 R b Replaced-C 1-10 alkyl;

[0124] The R 4 For -C 1-10 Alkyl; the C 1-10 The alkyl group is optionally substituted by 1, 2 or 3 Rc replace;

[0125] The R 5 is H;

[0126] The R b are independently halogen;

[0127] The R c are independently halogen;

[0128] The R 6 is cyano; said R 7 is H; said R 8 is a halogen; said n is 0;

[0129] Solution 5:

[0130] Said for

[0131] The R 3 is -H or oxo;

[0132] The R 1 -C(O)R 11 , -C 1-4 Alkyl, -C 3-6 Cycloalkyl, phenyl or "a 5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen"; the -C 1-4 Alkyl, the -C 3-6 The cycloalkyl group, the phenyl group and the “5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen” are independently optionally substituted by 1, 2 or 3 R a replace;

[0133] The R 11 is -CF3, cyclopropyl or phenyl;

[0134] The R 2 is H;

[0135] The R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted by 1, 2 or 3 R c replace;

[0136] The R 5 is H;

[0137] The R 6 is cyano; said R 7 is H; said R 8 is halogen; said n is 0; 1

[0138] Solution 6:

[0139] Said for

[0140] The R 3 is -H or oxo;

[0141] The R 1 -C(O)R 11 , phenyl or "a 5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen"; the phenyl group and the "a 5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen" are independently optionally replaced by 1, 2 or 3 R a replace;

[0142] The R 11 For -C 3-6 Cycloalkyl;

[0143] The R 2 is H;

[0144] The R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted with 1 cyclopropyl group;

[0145] The R 5 is H;

[0146] The R 6 is cyano; said R 7 is H; said R 8 is halogen; and n is 0.

[0147] In a preferred embodiment, the compound as shown in formula I is selected from any of the following compounds:

[0148]

[0149]

[0150] In a preferred embodiment, the compound as shown in formula I is selected from any of the following compounds:

[0151]

[0152]

[0153]

[0154]

[0155] The present invention also provides a method for preparing the compound shown in formula I, which comprises the following steps: subjecting the compound shown in formula II to a dehydration reaction to obtain the compound shown in formula I;

[0156]

[0157] Among them, X 1 , X 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 As described in any one of the present invention, R 9 for

[0158] The present invention also provides a compound as shown in formula II,

[0159] Among them, X 1 , X 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 and R 9 As described in any one of the present invention.

[0160] In a preferred embodiment, the compound as shown in formula II is selected from any of the following compounds:

[0161]

[0162]

[0163] The present invention also provides a compound as shown in formula III in, for R X2 , R X3 , R 1 , R 2 , R 3 , R 4 and R 5 As described in any one of the present invention.

[0164] In a preferred embodiment, the compound represented by formula III is selected from any one of the following compounds:

[0165]

[0166] The present invention also provides a pharmaceutical composition, which comprises the compound shown in Formula I or a pharmaceutically acceptable salt thereof (eg, an effective therapeutic amount of a pharmaceutically active ingredient), and a pharmaceutically acceptable carrier.

[0167] The present invention also provides a use of a compound as shown in Formula I, a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition in the preparation of a 3CLpro protease inhibitor (for example, in vivo or in vitro (such as a positive control in laboratory research)).

[0168] The present invention also provides a use of a compound as shown in Formula I, a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition in the preparation of a drug for treating and / or preventing diseases caused by coronavirus infection.

[0169] In a preferred embodiment, the coronavirus is one or more of 229Eα coronavirus, NL63α coronavirus, OC43β coronavirus, HKU1β coronavirus, Middle East respiratory syndrome coronavirus (MERS-cov), severe acute respiratory syndrome coronavirus (SARS-cov) and SARS-cov-2;

[0170] More preferably, the coronavirus is SARS-CoV-2.

[0171] The present invention also provides a use of a compound as shown in Formula I, a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical composition in the preparation of a drug for preventing and / or treating the 2019 coronavirus disease (COVID-19).

[0172] The present invention also provides a method for treating diseases related to coronavirus infection, which comprises applying the compound shown in the above formula I to a subject in need of such treatment (preferably, applying a therapeutically effective amount of the compound shown in the above formula I to a subject in need of such treatment), or a pharmaceutically acceptable salt thereof, or the above pharmaceutical composition.

[0173] Definition of terms

[0174] Unless otherwise specified, the terms used in the present invention have the following definitions. Definitions of terms not mentioned below are as commonly understood by those skilled in the art to which the present invention belongs.

[0175] The term "pharmaceutically acceptable salt" refers to a salt prepared from a compound with a relatively nontoxic, pharmaceutically acceptable acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of a pharmaceutically acceptable base in a pure solution or a suitable inert solvent. When the compound of the present invention contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of a pharmaceutically acceptable acid in a pure solution or a suitable inert solvent. When the compound contains relatively acidic and relatively basic functional groups, it can be converted into a base addition salt or an acid addition salt.

[0176] The end of the structural fragment It means that the structural fragment is connected to the rest of the molecule through this site. For example, It refers to cyclopropyl.

[0177] With wedge-shaped and dotted keys Indicates the absolute configuration of a stereocenter.

[0178] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0179] The term "oxo" refers to =O or -O - , oxygen atoms replace two hydrogens or lone pairs of electrons on the same carbon or nitrogen atom, in the form of =O or -O - .

[0180] For example

[0181] The term "alkyl" refers to a saturated, straight-chain or branched, monovalent hydrocarbon radical having a specified number of carbon atoms. 1-10 Alkyl refers to an alkyl group having 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) carbon atoms, for example, C 1-6 Alkyl, for example C 1-4 Alkyl. Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and the like.

[0182] The term "cycloalkyl" refers to a saturated, monocyclic hydrocarbon radical having the specified number of carbon atoms, for example, C 3-10 Cycloalkyl refers to a cycloalkyl group having 3-10 (e.g., 3, 4, 5, 6, 7, 8, 9 or 10) ring carbon atoms, such as C 3-6 Specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0183] The term "heterocyclyl" refers to a saturated, monocyclic, cyclic hydrocarbon substituent having a specified number of ring atoms (e.g., 3-10 members), a specified number of heteroatoms (e.g., 1, 2, 3, 4, or 5), and a specified type of heteroatom (one or more of N, O, and S). The heterocyclyl is attached to the rest of the molecule through a carbon atom or a heteroatom; the heterocyclyl is attached to the rest of the molecule through a ring with heteroatoms or a ring without heteroatoms. For example, a 3-10 membered heterocyclyl.

[0184] The term "aryl" refers to a radical having the specified number of carbon atoms (e.g., 6-10 ) is a cyclic, aromatic, monovalent hydrocarbon group, which is a single ring or a bicyclic ring (for example, 2). When it is a bicyclic ring, the single rings share two atoms and a bond. For example, C 6-10 Aryl, specific examples include but are not limited to oxyphenyl or naphthyl.

[0185] The term "heteroaryl" refers to a monocyclic, aromatic, monovalent group having a specified number of ring atoms (e.g., 5-10 members), a specified number of heteroatoms (e.g., 1, 2, 3, 4, or 5), a specified heteroatom type (one or more of N, O, and S). The heteroaryl group is attached to the rest of the molecule through a carbon atom or a heteroatom. For example, a 5-10 membered heteroaryl group, and another example is a 5-6 membered heteroaryl group with 1 or 2 heteroatoms being nitrogen. Specific examples include, but are not limited to: ).

[0186] In general, the term "substituted by..." means that one or more hydrogen atoms in a given structure are replaced by a specific substituent. Further, when the group is substituted by more than one of the substituents, the substituents are independent of each other, that is, the more than one substituents may be different or the same. Unless otherwise indicated, a substituent group may be substituted at each substitutable position of the substituted group. When more than one position in the given structural formula can be substituted by one or more substituents selected from a specific group, the substituents may be substituted at each position in the same or different manner.

[0187] The expression "a group B optionally substituted by one or more groups A" means that the group B may be unsubstituted or substituted by one or more groups A. For example, 1-6 The alkyl group is optionally substituted by 1, 2 or 3 R 3-2 Substitution refers to C 1-6 The alkyl group may be unsubstituted or substituted with 1, 2 or 3 R 3-2 replaced.

[0188] When a substituent is listed without indicating the atom through which it is attached to a compound included in the chemical formula but not specifically mentioned, such substituent may be bonded via any atom thereof. Combinations of substituents and / or variations thereof are permissible only if such combinations result in stable compounds.

[0189] When a group is listed without explicitly indicating that it has a substituent, such group is only meant to be unsubstituted. For example, when "C 1-6 When there is no limitation of "unsubstituted or substituted" before "alkyl", it only refers to "C 1-6 Alkyl" itself or "unsubstituted C 1-6 alkyl".

[0190] In addition, it should be noted that, unless otherwise explicitly stated, the description method "... independently are" used in the present invention should be understood in a broad sense, meaning that the individuals described are independent of each other and can independently be the same or different specific groups. In more detail, the description method "... independently are" can mean that in different groups, the specific options expressed by the same symbols do not affect each other; it can also mean that in the same group, the specific options expressed by the same symbols do not affect each other.

[0191] The term "one or more" or "one or more than two" means 1, 2, 3, 4, 5, 6, 7, 8, 9 or more.

[0192] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) ameliorating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0193] The term "prevention" refers to reducing the risk of developing a disease.

[0194] The term "therapeutically effective amount" refers to an amount of a compound that is sufficient to effectively treat or prevent a disease or condition described herein when administered to a patient. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, but can be adjusted as needed by those skilled in the art. Doses outside this range may also be used depending on the dosage form and the severity of the disease.

[0195] The term "subject" refers to any animal, usually a mammal, such as a human, that needs to be treated or prevented. Mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc.

[0196] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0197] The reagents and raw materials used in the present invention are commercially available.

[0198] The positive and progressive effects of the present invention are: in the applications disclosed in the present invention, the compounds thereof have equivalent or better main protease activity inhibition ability and better oxidase tolerance ability, achieve good anti-coronavirus activity and excellent human liver microsome stability, do not need to be used in conjunction with ritonavir etc. in clinical use, have a longer oxidative metabolism time in the body, reduce the number of oral administrations per day, and are conducive to improving drug compliance. DETAILED DESCRIPTION

[0199] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0200] Test conditions:

[0201] Prep-HPLC test parameters:

[0202] Instrument: Agilent;

[0203] Analytical column: Xtimate C18, 5um, 4.6*150mm;

[0204] Mobile phase: A: 0.05% TFA in H2O, B: 0.05% TFA in ACN;

[0205] Wavelength: 214nm / 254nm;

[0206] Flow rate: 1ml / min.

[0207] Chiral-HPLC test parameters:

[0208] Chromatographic column: Daicel IH 250*4.6mm,5μm;

[0209] Mobile phase A: HEX + 0.2% DEA;

[0210] Mobile phase B: IPAMEOH + 0.2% DEA;

[0211] Detection wavelength: 254nm;

[0212] Flow rate: 1 mL / min;

[0213] Injection volume: 10 μL;

[0214] Column temperature: RT;

[0215] Running time: 14 minutes;

[0216] Isocratic elution program: mobile phase A:mobile phase B=80:20 (V / V).

[0217] Example 1 Synthesis of intermediates

[0218] Example 1.1 Synthesis of intermediate s1

[0219]

[0220] Step 1:

[0221]

[0222] To a solution of s1-1 (67.0 g, 328 mmol) and NaOH (13.1 g, 328 mmol) in H2O (800 mL) was added a 38% aqueous solution of HCHO (45 mL, 623 mmol). The mixture was stirred at 25 ° C for 3 hours. Then stirred at 110 ° C for 3 hours. LC-MS showed that the reaction was complete. HCl was added to the reaction mixture at 0 ° C to quench pH = 7, and then the mixture was precipitated to obtain a brown solid. The brown solid was washed with water, filtered and concentrated under reduced pressure to obtain s1-2 (51.3 g, 72.2%), a brown solid.

[0223] LCMS: m / z = 217.2 [M+H] + .

[0224] 1 H NMR: (400MHz, DMSO-d6) δ = 10.91 (s, 1H), 7.45 (d, J = 8.0Hz, 1H), 7.33 (d, J = 8.0Hz, 1H), 7.08 ( t,J=7.2Hz,1H),7.02-6.96(t,J=7.2Hz,1H),4.28-4.12(m,2H),3.62-3.58(m,2H),3.14(br dd,J=5.1,16.2Hz,2H),2.81(br dd,J=10.3,16.2Hz,1H).

[0225] Step 2:

[0226]

[0227] To a solution of s1-2 (51.3 g, 237 mmol) in MeOH (1000 mL) was added SOCl2 (98.7 g, 830 mmol) at 0 ° C. The mixture was stirred at 70 ° C for 4 hours. LC-MS showed that the reaction was complete. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was diluted with water and quenched by adding NaHCO3 at 0 ° C. It was then extracted with ethyl acetate, and the combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give s1-3 (43.3 g, 79.3%) as a white solid.

[0228] LCMS: m / z = 231.4 [M+H] + .

[0229] Step 3:

[0230]

[0231] To a solution of s1-3 (43.2 g, 188 mmol) in DCM (1000 mL) was added TEA (23.6 g, 233 mmol) and Boc2O (45.0 g, 206 mmol). The mixture was stirred at 25 ° C for 16 hours. LC-MS showed that the reaction was complete. HCl was added to the reaction mixture at 0 ° C to pH = 6 to quench, and then the mixture was diluted with water and extracted with DCM. The organic layers after extraction were combined, washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (0-11% ethyl acetate / petroleum ether gradient elution) to give s1-4 (60.2 g, 97%) as a white solid.

[0232] LCMS: m / z=331.1[M+H] + .

[0233] 1 H NMR(400MHz, CHLOROFORM-d)δ=8.42-7.78(m,1H),7.52(d,J=7.6Hz,1H),7.33(dd,J=2.5,7.9Hz,1H),7.20-7.16(m,2H),5.50-5.17( m,1H),4.98-4.77(m,1H),4.66-4.49(m,1H),3.64(d,J=8.2Hz,3H),3.45(d,J=15.5Hz,1H),3.19-3.05(m,1H),1.56(d,J=1.7Hz,9H).

[0234] Step 4:

[0235]

[0236] To a solution of s1-4 (31.0 g, 93.8 mmol) in THF (400 mL) and H2O (400 mL) was added NBS (16.7 g, 93.8 mmol) and HOAc (407 g, 6.78 mol). The mixture was stirred at -5 °C for 1 hour. LC-MS showed that the reaction was complete. Saturated Na2SO3 solution (200 mL) was added to the reaction mixture to quench, and then NaHCO3 solution was added to the mixture at 0 °C until pH = 7. The above mixture was diluted with water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure to give a white solid s1-5 (32.0 g, 98% yield). The configuration of compound s1-5 was determined with reference to WO2023009187.

[0237] LCMS: m / z=347.2[M+H] + .

[0238] 1 H NMR (400MHz, DMSO-d6) δ = 10.67 (s, 1H), 7.27-7.22 (m, 1H), 7.12-6.97 (m, 2H), 6.91 (d, J = 7.8Hz, 1H), 4.67-4 .57(m,1H),3.75-3.68(m,3H),3.59-3.47(m,2H),2.44-2.35(m,1H),2.31-2.19(m,1H),1.41-1.37(m,9H).

[0239] Step 5:

[0240]

[0241] To a solution of compound s1-5 (500 mg, 1.44 mmol) in MeOH (5 mL) in a sealed tube was added NH4OH (20.6 mL, 144 mmol), and the mixture was stirred at 80° C. for 16 hours. The mixture was concentrated to give compound s1-6 (425 mg, yield 94%) as a white solid.

[0242] LCMS: m / z (ES+) = 354.2 [M+Na] + .

[0243] Step 6:

[0244]

[0245] A solution of compound s1-6 (1 g, 3.00 mmol) in HCl / dioxane (10 mL) was stirred at 25° C. for 3 hours. The mixture was concentrated to give compound s1 (1.2 g, crude) as a white solid.

[0246] LCMS: m / z (ES+) = 232.0 [M+H] + .

[0247] 1 H NMR(400MHz, DMSO-d6)δ9.02(brs,1H),8.04(s,1H),7.77(s,1H),7.64(d,J=7.6Hz,1H),7.29–7.27(m,1H), 7.26–7.24(m,1H),7.02–6.98(m,1H),4.61(s,2H),3.46–3.43(m,1H),2.51–2.49(m,2H),2.26–2.25(m,2H).

[0248] Example 1.2 Synthesis of intermediate s2

[0249]

[0250] Step 1:

[0251]

[0252] Under N2, Burgess reagent (2.04 g, 8.50 mmol) was added to a solution of compound s1-6 (564 mg, 1.70 mmol) in THF (20 mL), and the mixture was stirred at 25 ° C for 3 hours. The mixture was poured into saturated NaHCO3 solution (20 mL), then extracted with ethyl acetate (10 mLx3). The organic phase was washed with brine (10 mL), then dried and concentrated with Na2SO4, and the resulting residue was purified by silica gel column chromatography to obtain compound s2-1 (444 mg, 83% yield), a white solid.

[0253] 1 H NMR (400MHz, CDCl3) δ8.74(s,1H),7.30(d,J=8.0Hz,1H),7.06(t,J=7.6Hz,1H),6.99(d,J=7.6Hz,2H),4.86(dt, J=47.2,7.8Hz,1H),3.89–3.70(m,2H),2.92–2.78(m,1H),2.54(dd,J=13.2,8.3Hz,1H),1.54(d,J=44.0Hz,9H).

[0254] Step 2:

[0255]

[0256] To compound s2-1 (120mg, 0.383mmol) in TFA (2mL) and ACN (2mL) solution, add anisole (124mg, 1.15mmol), then the solution is stirred at 25 ° C for 1 hour. The mixture is concentrated, and the residue is quenched with saturated NaHCO (20mL), then extracted with ethyl acetate (15mLx3). The organic phase is washed with brine (15mL), then Na2SO4 is dried and concentrated. The obtained residue is purified by silica gel column chromatography to obtain compound s2 (64mg, 78% yield), a white solid.

[0257] 1 H NMR (400MHz, CDCl3): δ8.23(s,1H),7.26–7.19(m,2H),7.07(td,J=7.6,0.8Hz,1H),6.94(d,J=7.8Hz,1H),4.42(dd, J=8.4,6.5Hz,1H),3.57(d,J=11.2Hz,1H),3.17(d,J=11.2Hz,1H),2.69(dd,J=13.6,6.3Hz,1H),2.56–2.52(m,1H).

[0258] Example 1.3 Synthesis of Intermediate 26a

[0259]

[0260] Step 1:

[0261]

[0262] Tf2O (9.26 g, 32.82 mmol) was added to a solution of 26-1 (4.0 g, 27.36 mmol) and 2,6-dimethylpyridine (5.86 g, 54.73 mmol) in DCM (20 mL) at 0°C. After addition, the mixture was stirred at 0°C for 0.5 h. The reaction mixture was extracted with ethyl acetate (50 mL*3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, then filtered and concentrated under low pressure. The crude product was purified by silica gel chromatography (PE: EtOAc = 1: 0 to 5: 1) to give a brown oily compound 26-2 (4.81 g, 63% yield).

[0263] Step 2:

[0264]

[0265] To a solution of 26-3 (2.29 g, 9.38 mmol) and 26-2 (3.91 g, 14.06 mmol) in MeCN (10 mL) was added K2CO3 (2.59 g, 18.75 mmol) at 25°C. The resulting mixture was stirred at 50°C for 1.5 h under N2. The reaction mixture was diluted with H2O (10 mL) and then extracted with ethyl acetate (50 mL*3). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, then filtered and concentrated under low pressure. The crude product was purified by silica gel chromatography (PE:EtOAc=10:1~PE:EtOAc=5:1) to give compound 26-4 (2.13 g, 5.72 mmol, 61%) as a brown oil.

[0266] MS (ESI+) m / z 373.10 [M+H] + .

[0267] Step 3:

[0268]

[0269] To a solution of 26-4 (290 mg, 778.70 μmol) in THF (5 mL) and water (1 mL) was added LiOH·H2O (100 mg, 2.38 mmol). The mixture was stirred at 25°C for 3 h. The mixture was then concentrated, diluted with water (10 mL), adjusted to pH = 3-4 with 2 mol / L HCl, and extracted with EtOAc (20 mL*2). The organic layers were combined and concentrated to give 26a as a yellow oil.

[0270] MS (ESI+) m / z 359.10 [M+H] + .

[0271] Intermediate 27a was synthesized by a method similar to the above method, adjusting compound 26-3 to That's it.

[0272]

[0273] LCMS: m / z=320[M+1] + .

[0274] Example 1.4 Synthesis of intermediate 3a

[0275]

[0276] Step 1:

[0277]

[0278] At 0°C, 3-1 (5.0 g, 38.7 mmol) was mixed with H2O (50 mL) and an aqueous solution of HBr (60 mL, 48 wt%), and a solution of sodium nitrite (4.5 g, 65.22 mmol) in H2O (10 mL) was added dropwise to the mixture, and the mixture was further stirred at 0°C for 30 min and heated to 25°C overnight. The residue was extracted with EtOAc, washed with brine, dried over MgSO4, and concentrated to give compound 3-2 (7.50 g crude product) as a yellow oil.

[0279] LC-MS[M+1] + =192.9,194.9.

[0280] Step 2:

[0281]

[0282] To a solution of 3-2 (7.5 g, 38.9 mmol) in EtOH (120 mL) was added SOCl2 (18.49 g, 155.4 mmol, 11.3 mL) at 0° C., and the mixture was further stirred for 1 h at 25° C. The solvent of the mixture was removed, and purified by flash silica gel chromatography (PE:EA=100:1) to give 3-3 (5.5 g, 64.0% yield) as a colorless oil.

[0283] LC-MS[M+1] + =220.9,222.9.

[0284] Step 3:

[0285]

[0286] At 0°C, NaH (1.13 g, 28.35 mmol) was added to a mixture of 3-4 (2.1 g, 18.90 mmol), 3-3 (5.43 g, 24.57 mmol) and DMF (20 mL), and stirred at 0°C for 1 h. The mixture was quenched with cold water, extracted with EA, and washed with brine. Compound 3-5 was purified by flash silica gel chromatography (PE:EA=1:1) as a white solid (3.4 g, 72% yield).

[0287] LC-MS[M+1] + =252.2.

[0288] Step 4:

[0289]

[0290] At 25°C in an oxygen atmosphere, 3-5 (500 mg, 2.0 mmol), cyclopropylboronic acid (854.6 mg, 10.0 mmol), pyridine-N-oxide (PNO, A mixture of 1,4-dihydro-1,4-dihydro-2-nitropropene (2 ...

[0291] LC-MS[M+1] + =292.10.

[0292] Step 5:

[0293]

[0294] To a solution of 3-6 (100 mg, 343.2 μmol) in THF (1 mL) and H2O (1 mL) was added LiOH·H2O (13.7 mg, 343.2 μmol). The mixture was stirred at 25°C for 1 h. The resulting mixture was extracted with EA, the aqueous layer was acidified to pH = 5 with HCl (2 mol / L), extracted with EA, the organic phase was washed with brine, dried over Na2SO4, and concentrated to give a mixture 3a (80.00 mg, crude) as a white solid.

[0295] LC-MS[M+1] + =264.2.

[0296] The following compounds were synthesized by methods similar to those described above:

[0297]

[0298]

[0299] Example 1.5 Synthesis of Intermediate 10a

[0300]

[0301] Step 1:

[0302]

[0303] To a solution of 10-1 (700 mg, 2.9 mmol) in DMSO (3 mL) was added cyclopropylamine (246.7 mg, 4.3 mmol) and DIEA (1.1 g, 8.6 mmol). The mixture was stirred at 130 ° C for 1 hour. The crude product was purified by reverse phase HPLC (ACN / H2O: 0-100%) to give compound 10-2 (520 mg, 82%) as a yellow solid.

[0304] LCMS: m / z = 220.2 [M+H] + .

[0305] Step 2:

[0306]

[0307] To a solution of 10-2 (300 mg, 1.4 mmol) in dioxane (10 mL) was added Cs2CO3 (1.3 g, 4.1 mmol) and 10-3 (368.5 mg, 1.8 mmol). The mixture was stirred at 80 ° C for 2 hours. The reaction mixture was extracted with EA, the organic phase was separated, washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give compound 10-4 (450 mg, crude), a yellow oil.

[0308] LCMS: m / z=346.0 [M+H] + .

[0309] Step 3:

[0310]

[0311] To a solution of 10-4 (450 mg, 1.3 mmol) in THF (2 mL) and H2O (2 mL) was added LiOH·H2O (109.4 mg, 2.6 mmol). The mixture was stirred at 25°C for 2 hours. The mixture was adjusted to pH = 4 with HCl (1 mol / L) and directly concentrated. The resulting product was purified by reverse phase HPLC (ACN / H2O: 0-50%) to give compound 10a (224.4 mg, 52%) as a yellow solid.

[0312] LCMS: m / z = 332.2 [M+H] + .

[0313] 1H NMR (400MHz, CHLOROFORM-d) δ = 7.04 (s, 1H), 6.41 (br s,1H),5.38(dd,J=5.6,9.6Hz,1H),2.92(qt,J=3.7,7.2Hz,1H),2.11-1.97(m,2H),0.92-0.86(m,2H),0.69-0.60(m,3H),0.51(br d,J=7.0Hz,2H),0.14(br dd,J=2.6,13.6Hz,2H).

[0314] The following compounds were synthesized by methods similar to those described above:

[0315] Intermediate 16a:

[0316] Yellow solid

[0317]

[0318] LCMS: m / z = 404.0 [M+H] + .

[0319] 1 H NMR(400MHz, DMSO-d6)δ:9.20(s,1H),7.88–7.80(m,2H),7.39(t,J=9.7Hz,1H),7.16(t,J =8.9Hz,1H),5.26(dd,J=5.2,10.2Hz,1H),2.16–2.09(m,1H),2.03–1.96(m,1H),0.60(br s,1H),0.35(ddd,J=4.7,8.4,13.3Hz,2H),0.15–0.08(m,1H),0.00–-0.06(m,1H).

[0320] Example 1.6 Synthesis of Intermediate 12a

[0321]

[0322] Step 1:

[0323]

[0324] To an aqueous solution (5 mL) of 12-1 (300 mg, 2.10 mmol) was added an aqueous solution (1.85 mL, 40%) of HBr, and then an aqueous solution (5 mL) of NaNO2 (173 mg, 2.51 mmol) was added to the mixture at 0°C. The mixture was stirred at 25°C for 16 hours. The reaction mixture was extracted with EA (30 mL*2), the organic phase was separated, washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain a yellow oil 12-2.

[0325] 1 H NMR (400MHz, DMSO-d6) δ: 14.62–12.21 (m, 1H), 4.25 (t, J = 7.3Hz, 1H), 2.50–2.32 (m, 1H), 2.03–1.99 (m, 2H), 1.88–1.55 (m, 6H).

[0326] Step 2:

[0327]

[0328] SOCl2 (116 mg, 977 μmol) was added to a solution of 12-2 (405 mg, 1.90 mmol) in MeOH (3 mL). The mixture was stirred at 70°C for 16 hours. The reaction mixture was then concentrated under reduced pressure to obtain 12-3 as a yellow oil.

[0329] Step 3:

[0330]

[0331] NaHMDS (1 mol / L, 24.7 mL) was added to a solution of 2,4-difluoroaniline (1.4 g, 10.9 mmol) in THF (80 mL) at 0 °C under N2. After 1 hour, a solution of 12-4 (2.0 g, 8.2 mmol) in THF (10 mL) was added to the mixture. The reactants were then stirred at 0 °C for 2 hours. The reaction mixture was quenched with saturated NH4Cl (50 mL) at 0 °C, and then the reaction mixture was diluted with H2O (50 mL). The two phases were separated, and the aqueous phase was extracted with EA (100 mL×3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. 12-5 (1.6 g, 67%) was obtained as a yellow solid by rapid silica gel chromatography.

[0332] LCMS: m / z = 292.3 [M+H] + .

[0333] Step 4:

[0334]

[0335] Cs2CO3 (839 mg, 2.60 mmol) and 12-3 (247 mg, 1.12 mmol) were added to a solution of 12-5 (250 mg, 859 μmol) in dioxane (5 mL). The mixture was stirred at 80 °C for 16 hours. The reaction mixture was extracted with EA (40 mL × 2), the organic phase was separated, washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue, which was purified by reverse phase HPLC to give 12a (115 mg, 32.1%) as a white solid.

[0336] LCMS: m / z = 418.2 [M+H] + .

[0337] 1 H NMR(400MHz, DMSO-d6)δ:13.83–12.83(m,1H),9.22–9.11(m,1H),7.92–7.82(m,1H),7.73(s,1H),7.40-7.36(m,1H),7.20–7.11(m,1H) ,5.17–5.08(m,1H),2.34–2.26(m,1H),2.25–2.15(m,2H),2.02–1.90(m,1H),1.87–1.72(m,3H),1.71–1.63(m,1H),1.58–1.47(m,1H).

[0338] The following compounds were synthesized by methods similar to those described above:

[0339]

[0340] LCMS: m / z = 372.3 [M+H] + .

[0341] 1 H NMR(400MHz,DMSO-d6)δ:9.99(s,1H),7.93(s,1H),7.75(s,1H),7.67(s,1H),7.58(s,1H),5.17–5.13(m,1H),3.75(s,3 H),2.05–1.98(m,1H),1.92–1.89(m,1H),0.50–0.46(m,1H),0.28–0.23(m,2H),0.05–0.02(s,1H),0.00–-0.11(m,1H).

[0342] Example 1.7 Synthesis of Intermediate 15a

[0343]

[0344] Step 1

[0345]

[0346] To a solution of 15-1 (5.00 g, 26.1 mmol) in MeCN (125 mL) was added N,O-bis(trimethylsilyl)acetamide ( 13.3 g, 65.1 mmol), and then the mixture was stirred at 80 ° C for 2 hours. Then CH3I (5.60 g, 39.1 mmol) was added to the mixture at 25 ° C, and the mixture was stirred at 80 ° C for 16 hours. The reaction mixture was extracted with EA. The organic phase was separated, washed with brine (50 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue, which was flash silica gel chromatography to give 15-2 (1.30 g, 24.0%) as a white solid.

[0347] LCMS: m / z = 205.9, 207.9 [M+H] + .

[0348] Step 2

[0349]

[0350] A solution of 15-2 (1.10 g, 5.50 mmol) in DMF (20 mL) was purged with N2 three times. NaH (329 mg, 8.20 mmol) was then added to the above mixture at 0°C and stirred for 30 min. A solution of 15-3 (1.70 g, 8.20 mmol) in DMF (10 mL) was then added to the above reaction mixture. The mixture was stirred at 90°C for 16 hours. NH4Cl (100 mL) was added to the mixture at 25°C to quench the reaction, and then extracted with EtOAc (100 mL×2). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under reduced pressure, and the residue was subjected to flash silica gel chromatography to give 15-4 (1.70 g, 93.0%) as a colorless oil.

[0351] LCMS: m / z=332.0,334.0[M+H] + .

[0352] Step 3

[0353]

[0354] 15-4 (500 mg, 1.50 mmol) was dissolved in cyclopropylamine (6 mL) and stirred for 2 hours at 80° C. The reaction mixture was cooled to room temperature and then filtered, and the filter cake was concentrated under reduced pressure to obtain 15-5 as a white oil.

[0355] LCMS: m / z=309.3[M+H] + .

[0356] Step 4

[0357]

[0358] To a solution of 15-5 (464 mg, 1.50 mmol) in DCE (5 mL) was added hydroxy(trimethyl)stannane ( 5.40 g, 30.1 mmol). The mixture was stirred at 80°C for 16 h. H2O (2 mL) was added to the mixture to quench it, and then filtered and concentrated under reduced pressure. The residue was purified by reverse phase HPLC (ACN:H2O: 0-20%) to give 15a (160 mg, 36.1%) as a white solid.

[0359] LCMS: m / z = 295.2 [M+H] + .

[0360] 1 H NMR(400MHz,DMSO-d6)δ:5.09–4.98(m,1H),3.53(s,3H),2.00–1.91(m,1H),1.85–1.75 (m,1H),0.83–0.60(m,2H),0.32–0.22(m,6H),0.06–0.00(m,1H),-0.05–-0.12(m,1H).

[0361] Example 1.8 Synthesis of Intermediate 18a

[0362]

[0363] Step 1

[0364]

[0365] Pd2(dba)3(137mg, 151μmol), Xantphos(87.1mg, 151μmol) and Cs2CO3(1.47g, 4.52mmol) were added to a solution of 15-4(500mg, 602μmol) and 1-methylpyrazole-4-amine(190mg, 1.96mmol) in dioxane(10mL). The mixture was charged into N2 and stirred at 100°C under N2 atmosphere for 16hr. The reaction mixture was extracted with EtOAc(150mL x 2). The organic phase was separated, washed with brine (30mL), dried over Na2SO4, filtered and concentrated under reduced pressure, and the residue was purified by flash silica gel chromatography to give 18-1(280mg, 53.4%) as a yellow solid.

[0366] LCMS: m / z = 349.2 [M+H] + .

[0367] Step 2

[0368]

[0369] To a mixed solution of 18-1 (267 mg, 766 μmol) in THF (2 mL), H2O (2 mL) and MeOH (2 mL) was added NaOH (184 mg, 4.60 mmol). The mixture was stirred at 0 °C for 2 hours. The pH of the reaction solution was adjusted to 4 with HCl (1 mol / L) and extracted with EtOAc (20 mL×3). The organic phase was separated, washed with 30 mL of brine, dried over Na2SO4, filtered and concentrated under reduced pressure, and the residue was purified by reverse phase HPLC (H2O) to give 18a (30 mg, 11.7%) as a white solid.

[0370] LCMS: m / z=335.3 [M+H] + .

[0371] 1 H NMR (400MHz, DMSO-d6) δ: 9.10 (s, 1H), 7.93 (s, 1H), 7.58 (s, 1H), 4.99 (dd, J = 4.9, 10.3Hz, 1H), 3.81 (s,3H),3.47(s,3H),2.08–1.93(m,2H),0.59–0.49(m,1H),0.30–0.22(m,2H),0.00–-0.20(m,2H).

[0372] The following compounds were synthesized by methods similar to those described above:

[0373]

[0374] Example 1.9 Synthesis of Intermediate 22a

[0375]

[0376] Step 1:

[0377]

[0378] 22-1 (3 g, 15.9 mmol), 4-amino-3-fluoro-benzonitrile (2.59 g, 19.1 mmol), Pd2(dba)3 (1.45 g, 1.59 mmol), Xantphos (918 mg, 1.59 mmol), Cs2CO3 (15.5 g, 47.6 mmol) and dioxane (30 mL) were mixed, degassed and purged with N2 for 3 times, and then the mixture was stirred at 80 ° C under N2 atmosphere for 3 hours. The reaction mixture was diluted with EA (30 mL) and H2O (40 mL), the yellow solid was separated and filtered, and the filter cake was concentrated under vacuum to give compound 22-2 (2.9 g, crude) as a yellow solid.

[0379] LCMS: m / z = 245.1 [M+H] + .

[0380] Step 2:

[0381]

[0382] To a solution of 22-2 (1.2 g, 4.91 mmol) in ACN (10 mL) was added TMSCl (1.07 g, 9.83 mmol) and NaI (1.47 g, 9.83 mmol). The mixture was stirred at 50 °C for 2 hours. The reaction mixture was filtered to obtain a residue. A brown solid 22-3 (1.3 g, crude product) was obtained.

[0383] LCMS: m / z = 231.0 [M+H] + .

[0384] Step 3:

[0385]

[0386] To a solution of 22-3 (500 mg, 2.17 mmol) in DMF (20 mL) was added 22-4 (576 mg, 2.39 mmol), NaI (325 mg, 2.17 mmol) and Cs2CO3 (1.42 g, 4.34 mmol). The mixture was stirred at 80 °C for 3 hours. The reaction mixture was extracted with EA (150 mL*2). The organic phase was washed with brine (150 mL), dried over Na2SO4, filtered and concentrated under reduced pressure, and the residue was flash chromatographed on silica gel to give a white solid 22-5 (290 mg, 34.2%).

[0387] LCMS: m / z = 391.2 [M+H] + .

[0388] Step 4:

[0389]

[0390] To a solution of 22-5 (270 mg, 691 μmol) in THF (10 mL) and H2O (2 mL) was added LiOH·H2O (58.1 mg, 1.38 mmol). The mixture was stirred at 25°C for 20 min. The mixture was extracted with EA. The aqueous layer was separated, 1 mol / L HCl was added to adjust the pH of the aqueous layer to 5-6, and then extracted with EA again. The organic layers were combined, washed with brine (60 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give 22a (230 mg, crude) as a yellow solid.

[0391] LCMS: m / z = 363.2 [M+H] + .

[0392] The following compounds were synthesized by methods similar to those described above:

[0393]

[0394] Example 2 Synthesis of Compounds

[0395] Synthesis of compound 26:

[0396]

[0397] Step 1:

[0398] To a solution of 26a in DMF (4 mL) at 0°C, s1 (200.00 mg, 579.25 μmol, TFA), HATU (413.78 mg, 1.09 mmol) and NMM (256.84 mg, 2.54 mmol) were added, and the mixture was stirred at 0°C for 2 h. The mixture was quenched with NH4Cl (10 mL, aq, sat.), extracted with ethyl acetate (10 mL*2), and the organic layer was concentrated and purified by silica gel column chromatography (petroleum ether:ethyl acetate=10 / 1 to 0 / 1) to obtain 26a-1.

[0399] MS (ESI+) m / z = 594.10 [M + Na] + .

[0400] Step 2:

[0401] 5% Pd / C (100 mg, 227.42 μmol, 5% purity) was added to a solution of 26a-1 (130 mg, 227.42 μmol) in THF (8 mL). The mixture was degassed three times and then stirred at 25 °C under H2 atmosphere for 1 h. The mixture was filtered and concentrated to give 26a-2 (100.00 mg, crude) as a yellow solid, which was used in the next step without purification.

[0402] MS (ESI+) m / z = 460.20 [M + Na] + .

[0403] Step 3:

[0404] TFAA (216.04 mg, 1.03 mmol, 142.98 μL) and TEA (208.17 mg, 2.06 mmol, 286.73 μL) were added to a solution of 26a-2 (90 mg, 205.72 μmol) in DCM (5 mL). The mixture was stirred for 3 h at 30 ° C. The mixture was washed with water (5 mL * 3). The organic layer was concentrated and purified by prep-HPLC to obtain compound 26 (9.2 mg) as a white solid.

[0405] MS (ESI+) m / z = 538.10 [M + Na] + .

[0406] 1H NMR(400MHz, DMSO-d6)δ10.72(d,J=9.9Hz,1H),10.39(s,0.4H),10.06(s,0.6H),7.97-7.93(m,1H),7.74-7.63(m 1H),7.29-7.01(m,2H),6.91-6.78(m,2H),6.52-6.44(m,1H),5.78-5.73(m,1H),5.20-5.12(m,1H),4.08-3 .62(m,2H),2.68-2.59(m,1H),2.47-2.43(m,1H),2.09-1.77(m,2H),1.39-1.28(m,1H),0.96-0.69(m,6H).

[0407] Synthesis of compound 11:

[0408]

[0409] Step 1:

[0410]

[0411] To a solution of 11a (60 mg, 161.5 μmol) and s1 (37.3 mg, 161.5 μmol) in DMF (2 mL) was added NMM (65.3 mg, 646.3 μmol) and HATU (122.8 mg, 323.1 μmol). The reaction mixture was stirred at 25 °C for 1 h. The reaction was then quenched with water (10 mL) and extracted with EtOAc (10 mL×3). The organic phases were combined, washed with brine, dried over Na2SO4, filtered and concentrated. The residue was purified by flash silica gel chromatography (DCM: MeOH = 10: 1) to obtain 11a-1 (80.0 mg, 80%) as a white solid.

[0412] LC-MS[M+1] + =585.3.

[0413] Step 2:

[0414]

[0415] To a solution of 11a-1 (75.0 mg, 128.3 μmol) in DCM (5 mL) was added Berns reagent (306.6 mg, 1.2 mmol), and the mixture was stirred for 2 h at 25° C. The solvent was removed under reduced pressure and the residue was purified by prep-HPLC to give 11-P1 (11 mg, 15%) as a white solid and 11-P2 (13 mg, 18%) as a white solid.

[0416] Among them, one is compound 11-P1 and the other is compound 11-P2.

[0417] 11-P1

[0418] LC-MS: m / z = 567.0 [M+1] + .

[0419] 1H NMR(400MHz,DMSO-d6)δ:10.69(s,1H),9.97(s,1H),7.98(s,1H),7.71(s,1H),7.51(s,1H), 7.13(td,J=7.6,1.2Hz,1H),6.93(d,J=7.2Hz,1H),6.86–6.80(m,2H),5.72(d,J=8.0Hz,1H), 5.23–5.19(m,1H),4.12–4.09(m,2H),3.82(s,3H),2.67–2.60(m,1H),2.52–2.47(m,1H),2. 07–1.86(m,2H),0.70–0.62(m,1H),0.44–0.31(m,2H),0.22–0.16(m,1H),0.10–0.04(m,1H).

[0420] 11-P2

[0421] LC-MS: m / z = 567.0 [M+1] + .

[0422] 1H NMR(400MHz, DMSO-d6)δ:10.71(s,1H),10.15(s,1H),8.02(s,1H),7.75(s,1H),7.44(s,1H),7.26(td,J=7.6,8.0 Hz,1H),7.19(d,J=7.2Hz,1H),7.00(td,J=7.2,1.2Hz,1H),6.90(d,J=8.0Hz,1H),5.57(d,J=8.0Hz,1H),5.23–5. 19(m,1H),4.03(d,J=10.4Hz,1H),3.94(d,J=10.4Hz,1H),3.83(s,3H),2.68–2.62(m,1H),2.52–2.44(m,1H),2.0 4–1.97(m,1H),1.89–1.80(m,1H),0.66–0.60(m,1H),0.36–0.30(m,1H),0.21–0.08(m,2H),0.008–0.005(m,1H).

[0423] The following compounds were synthesized by methods similar to those described above:

[0424] Compound 12: synthesized from compound 12a.

[0425] LC-MS: m / z = 613.5 [M+1] + .

[0426] 1 H NMR(400MHz,DMSO-d6)δ:10.72–10.71(m,1H),9.24(s,0.5H),9.05(s,0.5H),7. 88–7.73(m,1H),7.64(s,0.6H),7.58(s,0.4H),7.41–7.34(m,1H),7.20–7.05(m, 2.8H),6.94–6.80(m,2.2H),5.57–5.47(m,1H),5.23–5.18(m,1H),4.08–3.89(m, 2H),2.69–2.59(m,1H),2.48–2.44(m,1H),2.22–2.08(m,3H),2.04–1.49(m,6H).

[0427] Chiral HPLC: The ratio of a pair of isomers is ~1:1.

[0428] There are two stereoisomers of compound 12.

[0429] Compound 13: synthesized from compound 13a.

[0430] Compound 13 was purified by Prep-HPLC to obtain the following two isomers.

[0431] Among them, one is compound 13-P1 and the other is compound 13-P2.

[0432] 13--P1

[0433] LC-MS: m / z = 530.4 [M+1] + .

[0434] 1H NMR(400MHz,DMSO-d6)δ:10.65(s,1H),9.27(s,1H),7.86(s,1H),7.53(s,1H),7.24 –7.07(m,1H),6.93–6.81(m,3H),5.89–5.45(m,1H),5.05–4.98(m,1H),3.79(s,4H), 3.73–3.68(m,1H),3.40(s,3H),2.67–2.60(m,1H),2.46–2.33(m,1H),2.11–1.82(m ,2H),0.66–0.56(m,1H),0.38–0.22(m,2H),0.09–0.03(m,1H),-0.12–-0.15(m,1H).

[0435] 13-P2

[0436] LC-MS: m / z = 530.4 [M+1] + .

[0437] 1 H NMR(400MHz, DMSO-d6)δ:10.71(s,1H),9.37(s,1H),7.94(s,1H),7.57(s,1H),7.26(td,J=7.6,1. 2Hz,1H),7.03–6.96(m,2H),6.90(d,J=7.6Hz,1H),5.37–5.31(m,1H),5.23–5.19(m,1H),3.81–3.7 8(m,3H),3.50(s,3H),3.41–3.36(m,2H),2.40–2.32(m,1H),2.24–2.16(m,1H),2.02–1.95(m,1H), 1.63–1.56(m,1H),0.66–0.60(m,1H),0.34–0.20(m,2H),0.06–0.016(m,1H),-0.20–-0.25(m,1H).

[0438] Compound 14: synthesized from compound 14a.

[0439] Compound 14 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0440] Among them, one is compound 14-P1 and the other is compound 14-P2.

[0441] 14-P1

[0442] LC-MS: m / z = 562.3 [M+1] +.

[0443] 1 H NMR(400MHz,DMSO-d6)δ:10.73(s,1H),8.29(s,1H),7.83–7.70(m,1H),7.37–7.24(m,2 H),7.11–6.90(m,4H),5.37–5.31(m,1H),5.25(t,J=8.4Hz,1H),3.55(d,J=10.4Hz,1H) ,3.41–3.36(m,4H),2.57–2.54(m,1H),2.43–2.37(m,1H),2.27–2.20(m,1H),1.62–1.5 4(m,1H),0.68–0.59(m,1H),0.36–0.20(m,2H),0.06–0.02(m,1H),-0.15–-0.22(m,1H).

[0444] 14-P2

[0445] LC-MS: m / z = 562.3 [M+1] + .

[0446] 1 H NMR(400MHz,DMSO-d6)δ:10.68(s,1H),8.27(s,1H),7.77–7.65(m,1H),7.36–7.31(m,1H),7 .26–7.06(m,2H),6.94–6.84(m,3H),5.51-5.47(m,1H),5.05(t,J=8.0Hz,1H),3.70(d,J=10 .8Hz,1H),3.57(d,J=10.4Hz,1H),3.30(s,3H),2.61–2.53(m,1H),2.46–2.41(m,1H),2.11– 1.85(m,2H),0.70–0.64(m,1H),0.35–0.30(m,2H),0.13–0.06(m,1H),-0.07–-0.13(m,1H).

[0447] Compound 15: synthesized from compound 15a.

[0448] Compound 15 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0449] Among them, one is compound 15-P1 and the other is compound 15-P2.

[0450] 15-P1:

[0451] LC-MS: m / z = 490.30 [M+H] + .

[0452] 1 H NMR(400MHz,CD3OD)δ:7.28–7.21(m,1H),7.07–6.90(m,3H),5.57–5.53(m,1 H),5.44–5.32(m,1H),3.51–3.45(m,2H),3.39(s,3H),2.62(d,J=8.0Hz,1H), 2.48–2.32(m,1H),2.31–2.23(m,1H),2.06–2.00(m,1H),1.78–1.58(m,1H),0 .75–0.62(m,3H),0.59–0.25(m,4H),0.11–0.03(m,1H),-0.08–-0.18(m,1H).

[0453] 15-P2:

[0454] LC-MS: m / z = 490.30 [M+H] + .

[0455] 1 H NMR(400MHz,CD3OD)δ:7.30–7.23(m,1H),7.07–6.94(m,3H),5.43–5.32( m,1H),5.25–5.19(m,1H),3.66–3.60(m,1H),3.51–3.45(m,4H),2.63–2. 52(m,2H),2.38–2.00(m,2H),1.65–1.57(m,1H),0.91–0.88(m,1H),0.75 –0.51(m,4H),0.41–0.26(m,2H),0.15–0.03(m,1H),-0.13–-0.18(m,1H).

[0456] Compound 17: synthesized from compound 17a.

[0457] Compound 17 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0458] Among them, one is compound 17-P1 and the other is compound 17-P2.

[0459] 17-P1

[0460] LC-MS: m / z = 580.0 [M+1] + .

[0461] 1 H NMR(400MHz,DMSO-d6)δ:10.69(s,1H),8.56(s,1H),7.31–7.17(m,3H),7.00–6.8 6(m,3H),5.56–5.48(m,1H),5.10–5.04(m,1H),3.71(d,J=10.4Hz,1H),3.58(d,J =10.8Hz,1H),3.21(s,3H),2.67–2.57(m,1H),2.47–2.41(m,1H),2.12–1.88(m,2 H),0.68–0.54(m,1H),0.40–0.24(m,2H),0.12-0.06(m,1H),-0.10–-0.19(m,1H).

[0462] 17-P2

[0463] LC-MS: m / z = 580.0 [M+1] + .

[0464] 1 H NMR (400MHz, DMSO-D6) δ10.74(s,1H),8.63(s,1H),7.31–7.24(m,3H),7.04–6.97(m,2H),6. 91(d,J=8.0Hz,1H),5.40–5.36(m,1H),5.23(t,J=8.8Hz,1H),3.51(d,J=10.4Hz,1H),3.41( d,J=10Hz,1H),3.26(s,3H),2.59–2.52(m,1H),2.43–2.38(m,1H),2.28–2.22(m,1H),1.63– 1.55(m,1H),0.63–0.59(m,1H),0.34–0.21(m,2H),0.07–0.01(m,1H),-0.20–-0.27(m,1H).

[0465] Compound 18: synthesized from compound 18a.

[0466] Compound 18 was purified by Prep-HPLC to obtain the following two isomers.

[0467] Among them, one is compound 18-P1 and the other is compound 18-P2.

[0468] 18-P1:

[0469] LC-MS: m / z = 530.30 [M+H] + .

[0470] 1H NMR(400MHz,DMSO-d6)δ:10.66(s,1H),8.72(s,1H),7.59(s,1H),7.25–7.13(m,1H),7.00–6.80 (m,3H),6.39(s,1H),5.49–5.36(m,1H),5.09–4.99(m,1H),3.74(s,3H),3.68(d,J=11.2Hz,1H), 3.56(d,J=11.2Hz,1H),3.35(s,3H),2.60–2.54(m,1H),2.47–2.40(m,1H),2.20–2.03(m,1H),1 .91–1.81(m,1H),0.68–0.57(m,1H),0.39–0.23(m,2H),0.11–0.03(m,1H),-0.10–-0.18(m,1H).

[0471] 18-P2:

[0472] LC-MS: m / z = 530.30 [M+H] + .

[0473] 1H NMR (400MHz, DMSO-d6) δ: 10.72 (s, 1H), 8.70 (s, 1H), 7.60 (d, J = 2.4Hz, 1H), 7.26 (d, J = 7.2Hz, 1H), 7. 03–6.89(m,3H),6.46(d,J=2.4,1H),5.36–5.32(m,1H),5.23–5.19(m,1H),3.74(s,3H),3.49(d,J=10 .4Hz,1H),3.44(s,3H),3.38(d,J=10.4Hz,1H),2.57–2.52(m,1H),2.43–2.32(m,1H),2.25–2.18(m,1 H),1.62–1.54(m,1H),0.66–0.58(m,1H),0.34–0.22(m,2H),0.06–0.01(m,1H),-0.18–-0.24(m,1H).

[0474] Compound 19: synthesized from compound 19a.

[0475] Compound 19 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0476] Among them, one is compound 19-P1 and the other is compound 19-P2.

[0477] 19-P1:

[0478] LC-MS: m / z = 580.4 [M+H] + .

[0479] 1 H NMR(400MHz, CD3OD)δ:8.15–7.94(m,1H),7.32–7.13(m,2H),7.09–6.87(m,3H),5.94–5.87(m,0. 2H),5.63–5.59(m,0.8H),5.19–5.03(m,1H),3.83(d,J=10.8Hz,1H),3.68–3.57(m,1.6H),3.46(s ,2.4H),2.89–2.77(m,0.2H),2.63(d,J=8.4Hz,1.8H),2.33–2.26(m,1H),1.86–1.77(m,0.8H),1 .75–1.66(m,0.2H),0.78–0.68(m,1H),0.46–0.33(m,2H),0.14–0.08(m,1H),0.00–-0.07(m,1H).

[0480] 19-P2:

[0481] LC-MS: m / z = 580.4 [M+H] + .

[0482] 1 H NMR(400MHz,CD3OD)δ:8.15–8.08(m,1H),7.31–7.24(m,2H),7.08–6.97(m ,2H),6.94(d,J=8.0Hz,1H),5.49–5.46(m,1H),5.24(t,J=8.8Hz,1H),3.62 –3.52(m,5H),2.65–2.53(m,2H),2.42–2.36(m,1H),1.69–1.57(m,1H),0.7 7–0.67(m,1H),0.44–0.26(m,2H),0.11–0.01(m,1H),-0.09–-0.16(m,1H).

[0483] Compound 20: synthesized from compound 20a.

[0484] Compound 20 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0485] Among them, one is compound 20-P1 and the other is compound 20-P2.

[0486] 20-P1:

[0487] LC-MS: m / z=518.2[M+H] + .

[0488] 1 H NMR(400MHz,CD3OD)δ:7.24–7.20(m,1H),7.04–6.94(m,2H),6.90(d,J=8.0Hz,1H),5.58 –5.53(m,1H),5.06–5.01(m,1H),3.80(d,J=10.8Hz,1H),3.60(d,J=10.4Hz,1H),3.46(s, 3H),2.69–2.56(m,2H),2.31–2.24(m,1H),2.00–1.87(m,1H),1.83–1.60(m,1H),0.98–0. 86(m,4H),0.76–0.67(m,1H),0.44–0.32(m,2H),0.12–0.06(m,1H),-0.03–-0.09(m,1H).

[0489] 20-P2:

[0490] LC-MS: m / z=518.3[M+H] + .

[0491] 1 H NMR(400MHz,CD3OD)δ:7.28(td,J=7.6,0.8Hz,1H),7.07–6.94(m,3H),5.44– 5.40(m,1H),5.23(t,J=8.8Hz,1H),3.61–3.47(m,5H),2.64–2.53(m,2H),2. 40–2.33(m,1H),1.96–1.89(m,1H),1.66–1.58(m,1H),0.99–0.87(m,4H),0. 76–0.66(m,1H),0.43–0.29(m,2H),0.09–0.03(m,1H),-0.07–-0.15(m,1H).

[0492] Compound 16: synthesized from compound 16a.

[0493] LC-MS: m / z = 599.3 [M+H] + .

[0494] 1H NMR(400MHz,DMSO-d6)δ:10.72(s,1H),9.27(s,0.5H),9.08(s,0.5H),7.81–7.74(m, 1H),7.67–7.59(m,1H),7.41–7.34(m,1H),7.28–7.12(m,3H),6.92–6.84(m,2H),5.7 2–5.59(m,1H),5.24–5.20(m,1H),4.11–3.97(m,2H),2.69–2.52(m,2H),2.11–1.82( m,2H),0.73–0.60(m,1H),0.48–0.31(m,1H),0.21–0.10(m,2H),-0.02–-0.04(m,1H).

[0495] Chiral HPLC: The ratio of one pair of isomers is ~55:45.

[0496] There are two stereoisomers of compound 16.

[0497] Compound 22: synthesized from compound 22a.

[0498] Compound 22 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0499] Among them, one is compound 22-P1 and the other is compound 22-P2.

[0500] LCMS::m / z=558.1[M+H] + .

[0501] 22-P1:

[0502] 1H NMR(400MHz,DMSO-d6)δ=10.73(br s,1H),8.58–8.46(m,2H),7.92(dd,J=1.8,11.3Hz,1H),7.73(dd,J=1.1,8.5Hz,1H),7.30(d, J=4.9Hz,1H),7.11(d,J=4.8Hz,1H),7.00–6.94(m,1H),6.81(d,J=7.8Hz,1H),6.73–6.60(m,2 H),5.87(dd,J=5.9,7.4Hz,1H),5.20(t,J=7.9Hz,1H),4.09(d,J=10.8Hz,1H),3.95(d,J=10.6 Hz,1H),2.65–2.58(m,1H),2.50–2.37(m,3H),1.36(d,J=18.1Hz,3H),1.31(d,J=18.0Hz,3H).

[0503] 22-P2:

[0504] 1 H NMR (400MHz, DMSO-d6) δ=10.77–10.53(m,1H),8.90–8.74(m,1H),8.60(t,J=8.4Hz,1H),7.93(dd,J =1.8,11.2Hz,1H),7.78–7.69(m,1H),7.33–7.19(m,3H),7.12(d,J=4.8Hz,1H),7.03(t,J=7.7Hz,1 H),6.91(d,J=7.7Hz,1H),5.82(dd,J=5.0,7.8Hz,1H),5.20(dd,J=6.4,8.8Hz,1H),3.94(d,J=10.6 Hz,1H),3.79(d,J=10.3Hz,1H),2.65(dd,J=8.8,13.4Hz,1H),2.49–2.34(m,3H),1.35–1.21(m,6H).

[0505] Compound 3: synthesized from compound 3a.

[0506] Compound 3 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0507] Among them, one is compound 3-P1 and the other is compound 3-P2.

[0508] 3-P1

[0509] LC-MS: m / z = 459.3 [M+1] +.

[0510] 1 H NMR(400MHz, DMSO-d6)δ10.73(s,1H),7.22–7.17(m,1H),6.87–6.79(m,5H),5.59(t,J=7.6Hz,1H),5.14-5.11(m,1H),4.01–3.66(m,2H),2.65–2 .60(m,2H),2.03–1.94(m,1H),1.88-1.81(m,2H),0.66–0.59(m,3H),0.5 2–0.44(m,2H),0.39–0.32(m,2H),0.19–0.10(m,1H),0.07–0.04(m,1H).

[0511] 3-P2

[0512] LC-MS: m / z = 459.30 [M+1] + .

[0513] 1 H NMR(400MHz,DMSO-d6)δ10.70(s,1H),7.28-7.24(m,1H),7.17-7.15(m,1H),7.03– 6.97(m,1H),6.93–6.84(m,3H),5.50(t,J=7.5Hz,1H),5.23-5.19(m,1H),4.01–3. 82(m,2H),2.75–2.63(m,2H),2.06–1.87(m,2H),1.82–1.70(m,1H),0.81–0.61(m, 5H),0.35–0.29(m,1H),0.24–0.20(m,1H),0.12–0.08(m,1H),-0.02–-0.07(m,1H).

[0514] Compound 4: Synthesized from compound 4a

[0515] LCMS: m / z = 513.0 [M+1] + .

[0516] 1H NMR(400MHz, DMSO-d6)δ10.72(s,1H),8.66(d,J=2.0Hz,0.5H),8.40(d,J=2.2Hz,0.5H),8.25-8.19(m,1H),7 .20-7.00(m,7H),6.99(d,J=1.9Hz,2H),5.68-5.58(m,1H),5.25-5.16(m,1H),4.16(d,J=10.7Hz,0.5H),4.02 (dd,J=20.0,10.6Hz,1H),3.82(d,J=10.6Hz,0.5H),2.67-2.61(m,1H),2.03-1.96(m,1H),1.92-1.85(m,1H) ,1.81-1.73(m,1H),1.23(s,1H),0.72-0.59(m,1H),0.48-0.30(m,1H),0.23-0.15(m,1H),0.13-0.06(m,1H).

[0517] Compound 8: Synthesized from compound 8a

[0518] Compound 8 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0519] Both are white solids, one of which is compound 8-P1 and the other is compound 8-P2.

[0520] 8-P1

[0521] LC-MS m / z=514.3[M+H] + .

[0522] 1 H NMR(400MHz,DMSO-d6)δ10.71(s,1H),8.99(s,1H),8.23-8.21(d,J=4.8Hz,1H),7.76-7 .71(m,1H),7.31-7.27(m,1H),7.21-7.17(m,2H)7.06-6.98(m,1H),6.94-6.83(m,3H), 5.66(t,J=7.6Hz,1H),5.20-5.15(m,1H),4.15-4.09(m,2H),2.68-2.59(m,1H),2.52-2 .49(m,1H),2.02-1.85(m,2H),0.72-0.72(m,1H),0.44-0.36(m,2H),0.22-0.16(m,2H).

[0523] 8-P2

[0524] LC-MS m / z=514.3[M+H] + .

[0525] 1 H NMR (400MHz, DMSO-d6) δ10.75(s,1H),9.25(s,1H),8.22(d,J=4.8Hz,1H),7.78–7.71(d,J=4.8Hz,1H),7.34–7.2 7(m,2H),7.20–7.16(m,1H),7.06–7.05(m,1H),7.02–6.98(m,1H)6.94–6.89(m,2H),5.60(t,J=8.0Hz,1H),5.25 –5.20(m,1H),4.01–3.99(m,1H),3.83–3.83(m,1H),2.69–2.62(m,1H),2.49–2.49(m,1H),2.03–1.96(m,1H),1. 82–1.73(m,1H),0.75–0.65(m,1H),0.37–0.30(m,1H),0.21–0.15(m,1H),0.11–0.07(m,1H),0.00–-0.05(m,1H).

[0526] Compound 5: Synthesized from compound 5a

[0527] LCMS: m / z = 531.0 [M+H] + .

[0528] 1 H NMR(400MHz,DMSO-d6)δ10.71(s,1H),8.74(s,0.6H),8.48(s,0.4H),8.03 -8.01(m,1H),7.12-7.01(m,4H),6.94-6.90(m,4H),5.62-5.60(m,1H),5.23-5.21(m,1H),4.13-3.98( m,2H),2.71-2.59(m,1H),2.46(m,1H),1.99-1.78(m,2H),0.85(m,1H),0.62-0.34(m,3H),0.19(m,1H).

[0529] Compound 7: Synthesized from compound 7a

[0530] LC-MS: m / z = 525.3 [M+H] + .

[0531] 1H NMR(400MHz,DMSO-d6)δ10.71(s,1H),9.57(s,1H),8.16(s,1H),7.65(s,1H),7.23–7.16(m,2H) ,7.02–6.98(m,2H),6.91–6.82(m,2H),5.59–5.54(m,1H),5.24–5.19(m,1H),3.97–3.94(m,1H) ,3.77–3.63(m,1H),2.67–2.61(m,1H),2.51–2.47(m,1H),2.02–1.96(m,1H),1.79–1.72(m,1H) ,0.96–0.91(m,5H),0.61–0.57(m,1H),035–0.29(m,1H),0.12–0.06(m,2H),0.03–-0.07(m,1H).

[0532] Compound 6: synthesized from compound 6a.

[0533] Compound 6 was purified by Prep-HPLC to obtain the following two stereoisomers.

[0534] Among them, one is compound 6-P1 and the other is compound 6-P2.

[0535] 6-P1

[0536] LCMS: m / z=499.30[M+1] + .

[0537] NMR: 1 H NMR(400MHz,DMSO-d6)δ10.69(s,1H),9.34(s,1H),8.07(s,1H),7.61(s,1H),7.16–7.06(m,1 H),6.95(s,2H),6.84(dq,J=15.4,7.28Hz,3H),5.67(t,J=7.6Hz,1H),5.16(dd,J=8.75,6.61H z,1H),4.07(s,2H),3.80(s,3H),2.63(dd,J=13.4,8.89Hz,1H),2.49-2.45(m,1H),1.94-1.86 (m,2H),0.71–0.58(m,1H),0.46–0.29(m,2H),0.17(dq,J=8.4,4.6Hz,1H),0.08–0.04(m,1H).

[0538] 6-P2

[0539] LCMS: m / z=499.30[M+1] + .

[0540] NMR: 1 H NMR(400MHz,DMSO-d6)δ10.63(s,1H),9.61(s,1H),8.08(s,1H),7.62(s,1H),7.23(td,J=7.70,1.09Hz,1 H),7.15(d,J=7.2Hz,1H),7.07–6.75(m,5H),5.57-5.50(m,1H),5.23-5.15(m,1H),3.92(d,J=7.2Hz,1H), 3.80-3.70(m,4H),2.64–2.57(m,1H),2.45–2.36(m,1H),1.99–1.82(m,1H),1.71(dt,J=14.97,7.90Hz,1 H), 0.56 (dq, J=9.05, 4.88Hz, 1H), 0.29 (tt, J=9.10, 4.04Hz, 1H), 0.19–0.03 (m, 2H), -0.04–-0.11 (m, 1H).

[0541] Compound 10: synthesized from compound 10a.

[0542]

[0543] LC-MS [M+H] + =527.30.

[0544] 1HNMR (400MHz, DMSO-d6) δ10.71–10.69(d,J=7.2Hz,1H),7.95–7.94(d,J=4.8Hz,0.5H),7.74–7.73(d,J=5.2Hz,0.5H),7.42( s,0.5H),7.38(s,0.5H),7.25–7.17(m,1.5H),7.01–6.97(m,0.5H),6.91–6.80(m,2H),5.66–5.59(m,0.5H),5.54-5.50(m,0. 5H),5.22–5.18(m,1H),4.07–3.39(m,1.5H),3.89–3.87(d,J=10.4Hz,0.5H),2.85–2.72(m,1H),2.67–2.59(m,1H),2.49–2. 46(m,1H),2.00–2.54(m,1H),1.89–1.76(m,1H),0.70–0.56(m,5H),0.45–0.28(m,2H),0.20–0.15(m,1H),0.10–0.04(m,1H).

[0545] Chiral HPLC: the ratio of one pair of isomers was 55%:45%.

[0546] There are two stereoisomers of compound 10.

[0547] Biochemical tests

[0548] Test 1: 3CLpro protease inhibition assay

[0549] The test substance (example compound) was diluted 3-fold in series with DMSO, with a total of 10 test points. The above solution was added to a 384-well microplate using an ECHO workstation, and each test point was tested in duplicate. 25 μL of 3CLpro protease solution was added to each well of the microplate containing the test substance. After the solution was mixed, it was pre-incubated at room temperature for 30 minutes. After the incubation was completed, 5 μL of substrate (Dabcyl-KTSAVLQ∥SGFRKM-E (Edans)) was added. The final concentrations of protease 3CLpro and substrate were 25 nM and 25 μM, respectively. For the 100% inhibition (HPE, hundred percent effect) control sample, 1 μM compound GC376 was added. For the 0% inhibition (ZPE, zero percent effect) control sample, no compound was added. The final concentration of the DMSO solution was 1%. Each test point was standardized using a background control to exclude background fluorescence interference. After incubation at 30°C for 60 minutes, the fluorescence signal (RFU) was detected at Ex / Em=340nm / 490nm using an ELISA reader M2e (SpectraMax). The inhibitory activity was calculated using the following formula: Inhibition %=[(sample-average ZPE) / (average HPE-average ZPE)]*100%.

[0550] 3CLpro protease solution: 20 mM Tris-HCl (pH 7.3), 100 mM NaCl, 1 mM EDTA, 5 mM TCEP and 0.1% BSA.

[0551] HEP: 100% inhibition control sample, containing substrate + 3CLpro protease solution + 1 μM GC376.

[0552] ZPE: 0% inhibition control sample, containing substrate + 3CLpro protease solution, without analyte.

[0553] Sample: Compound activity test sample. Contains substrate + 3CLpro protease solution + test substance.

[0554] Background control: contains the test substance + substrate, without protease.

[0555] Draw the inhibition curve and calculate the half inhibitory concentration IC 50 ,The results are shown in Table 1.

[0556] The compound of the present invention has 3CLpro protease inhibitory activity comparable to that of namatevir.

[0557] Test 2: In vitro SARS-CoV-2 replicon antiviral assay

[0558] The test substance was serially diluted 3 times with DMSO solution, with a total of 8 test points. 0.3 μL of the test substance solution was added to a 384-well microplate, and each test point was tested in duplicate. The RNA replicon was obtained by in vitro transcription (the in vitro transcribed RNA was purified using the RNeasy MinElute Cleanup Kit (QIAGEN 74204), OD260 / 280≈2, RNA Conc≥1000ng / uL). Huh7 cells transfected with purified SARS-CoV-2 replicon RNA (the constructed Replicon was derived from GenBank: NC045512, T7Promotor was added to the 5' end, the ORFs of the S and E genes were deleted, and the EGFP and bsd genes were replaced and inserted respectively. PolyA was retained at the 3' end, HDV ribozyme sequence and T7 Terminator were added, connected to the BAC vector (GENWIZ), and the linearized in vitro transcribed purified RNA was electrotransferred to the cells.) and inoculated at 4000 / well. In a 384-well microplate containing serial dilutions of compounds, the cells were then incubated for 1 day at 37°C and 5% CO2. The final volume of the cell culture was 60 μL per well and the final DMSO concentration in the assay plate was 0.5%.

[0559] Fluorescence intensity was measured using Acumen Cellista (TTP LabTech), and the antiviral activity of the compound was calculated based on the inhibition of GFP expression. Cell viability was measured using CellTiter Glo according to the manufacturer's instructions.

[0560] The antiviral activity and cell viability of the compounds were expressed as % inhibition and % viability, respectively, and were calculated using the following formula:

[0561] Inhibition rate (%) = (CPD-average ZPE) / (average HPE-average ZPE)*100

[0562] Cell viability (%) = (CPD - average HPE) / (average ZPE - average HPE) * 100

[0563] CPD: measured value of the sample containing the substance to be tested

[0564] ZPE: Mean value of virus control

[0565] HPE: average value of blank control (containing culture medium only).

[0566] The nonlinear four-parameter model was used to fit the natural log-transformed inhibition rate-response and calculate the EC 50 and CC 50 The results are shown in Table 1. The software used was GraphPad Prism.

[0567] The compound of the present invention has antiviral activity and cell viability comparable to those of namatevir.

[0568] Table 1

[0569] Compound No. <![CDATA[IC 50 (nM)]]> <![CDATA[EC 50 (nM)]]> <![CDATA[CC 50 (nM)]]> Nirmatrelvir A A >1000 1 A A >1000 2 A A >1000 3 B A >1000 4 A A >1000 5 A A >1000 6 A A >1000 8 A A >1000 10 A A >1000 11 A A >1000 12 A A >1000 13 A A >1000 14 A A >1000 15 A A >1000 16 A A >1000 17 A A >1000 18 A A >1000 19 A A >1000 20 A B >1000 22 A A >1000 24 A A >1000 25 A A >1000 26 A A >1000 27 B B >1000

[0570] Note: A≤100, B 100-1000, C>1000.

[0571] Test 3: Human liver microsome stability

[0572] The test substance was dissolved in DMSO to prepare a 10mM stock solution and diluted to 100μM with ACN:H2O=1:1 (v / v) to obtain a working solution. The working solution was diluted to 1μM in potassium phosphate buffer (100mM, pH 7.4) containing MgCl2 (3mM), NADPH (1mM) and HLM (1mg / mL, human liver microsomes) with a final volume of 200μL and incubated on a shaker at 37°C. The incubation was carried out in the presence and absence of the selective CYP3A inhibitor ketoconazole (1μM). At the selected time points (0, 5, 15, 30, 45, 60, 80min), 200μL of acetonitrile (containing 300ng / mL tolbutamide) was added to 100μL of the sample to terminate the incubation, shake and mix for 5min, and centrifuge at 3700rmp for 15min. The supernatant was transferred to a 96-well plate, diluted with mobile phase A (0.1% formic acid in water), and analyzed by liquid chromatography-mass spectrometry (LC-MS / MS). 1 / 2 Calculation formula t 1 / 2 =-0.693 / k, the data are shown in Table 2, where k is the slope of the linear fitting of the peak area ratio-time after natural logarithm transformation.

[0573] Compounds 1-25 of the present invention have better human liver microsomal stability (ie, oxidase stability) than nematevir, and compounds 26 and 27 have human liver microsomal stability (ie, oxidase stability) comparable to nematevir.

[0574] Table 2

[0575] Compound No. Human liver microsome half-life t1 / 2 (min) Nirmatrelvir C 5 B 12 B 13 B 14 A 16 B 17 B 18 A 20 A 22 B 26 C

[0576] Remarks: t1 / 2(min):A>100,B 30~100,C<30.

Claims

1. Use of a compound as shown in formula I or a pharmaceutically acceptable salt thereof in the preparation of a 3CLpro protease inhibitor or a drug: The medicine is a medicine for treating and / or preventing diseases caused by coronavirus infection; in, X 1 is N or CH; X 2 For-NR X2 , CR X3 or CR X4 R X5 ; R X2 , R X3 , R X4 and R X5 are independently -H, -C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl; R 3 H, oxo, halogen, C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl; is a single bond or a double bond; R 1 and R 2 are independently H, -C(O)R 11 、-C(O)OR 12 、-S(O) 2 R 13 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the "5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" and the "3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S" are independently optionally replaced by 1, 2 or 3 R a replace; R 11 , R 12 and R 13 Independently -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S", the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the "5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" and the "3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S" are independently optionally replaced by 1, 2 or 3 R b replace; R 4 and R 5 are independently H, -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R c replace; R a are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl; R b are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl; R c are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl; R 6 is cyano; R 7 H; R 8 is a halogen; n is 0 or 1; When X 1 CH, X 2 CR X3 or CR X4 R X5 , R 1 and R 2 One of the groups is H and the other is -C(O)R 11 、-C(O)OR 12 or -S(O) 2 R 13 , R 11 , R 12 and R 13 Independently -C 1-10 When the -C 1-10 The alkyl group is replaced by 1, 2 or 3 R a replace.

2. The use according to claim 1, It is characterized in that The coronavirus is one or more of 229Eα coronavirus, NL63α coronavirus, OC43β coronavirus, HKU1β coronavirus, Middle East respiratory syndrome coronavirus, severe acute respiratory syndrome coronavirus and SARS-cov-2; For example, the coronavirus is SARS-CoV-2.

3. The use according to claim 1 or 2, It is characterized in that The compound represented by formula I satisfies one or more of the following conditions: (1)R X2 and R X3 In the -C 1-6 The alkyl groups are independently -C 1-4 Alkyl groups, such as methyl; (2)R X2 and R X3 wherein the -C substituted by 1, 2 or 3 halogens 1-6 -C in the alkyl group 1-6 The alkyl groups are independently -C 1-4 Alkyl groups, such as methyl; (3)R 1 , R 2 , R 11 , R 12 , R 13 , R 4 , R 5 , R a , R b and R c In the -C 1-10 Alkyl and the -C substituted by 1, 2 or 3 halogens 1-6 -C in the alkyl group 1-6 The alkyl groups are independently -C 1-6 Alkyl, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or tert-butyl, for example methyl; (4)R 1 , R 2 , R 11 , R 12 and R 13 In the -C 3-10 The cycloalkyl groups are independently -C 3-6 cycloalkyl, such as cyclopropyl or cyclobutyl; (5)R a and R c In the -C 3-6 Cycloalkyl is independently cyclopropyl or cyclobutyl; (6)R 1 , R 2 , R 11 , R 12 and R 13 In the -C 6-10 Aryl is independently phenyl or naphthyl; (7)R 1 and R 2 In the above, the "heteroatoms selected from 1, 2 or 3 of N, O and S, 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms" is independently "heteroatoms selected from N, 5-10 membered heteroaryl group having 1 or 2 heteroatoms", preferably "heteroatoms selected from N, 5-6 membered heteroaryl group having 1 or 2 heteroatoms", more preferably pyridyl or diazolyl; The diazolyl group is preferably 1H-pyrazolyl; (8)R X2 , R X3 , R X4 , R X5 , R a , R b and R c wherein the halogen is F.

4. The use according to claim 1 or 2, It is characterized in that The compound represented by formula I satisfies one or more of the following conditions: (1) X 1 N or CH; (2) X 2 NR X2 or CR X3 ; (3) R X2 For -C 1-6 alkyl; (4) R X3 is H or -C substituted by 1, 2 or 3 halogens 1-6 alkyl; (5) R 3 is -H or oxo; (6) R 2 is H; (7) R 1 -C(O)R 11 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 Aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group and the "5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" are independently optionally replaced by 1, 2 or 3 R a replace; Preferably, the R 1 -C(O)R 11 , -C 1-4 Alkyl, -C 3-6 Cycloalkyl, phenyl or "a 5-6 membered heteroaryl group having 1 or 2 nitrogen atoms as heteroatoms"; the -C 1-4 Alkyl, the -C 3-6 The cycloalkyl group, the phenyl group and the "5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen" are independently optionally substituted by 1, 2 or 3 R a replace; More preferably, the R 1 -C(O)R 11 , phenyl or "a 5-6 membered heteroaryl group wherein the heteroatom is nitrogen and the number is 1 or 2"; the phenyl group and the "a 5-6 membered heteroaryl group wherein the heteroatom is nitrogen and the number is 1 or 2" are independently optionally replaced by 1, 2 or 3 R a replace; (8) R 11 For -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R b replace; Preferably, the R 11 For -C 1-4 Alkyl, -C 3-6 Cycloalkyl or phenyl; the -C 1-4 Alkyl, the -C 3-6 The cycloalkyl and phenyl groups are optionally substituted with 1, 2 or 3 halogens; for example, -CF 3 , cyclopropyl or phenyl; More preferably, the R 11 For -C 3-6 Cycloalkyl; (9) R 4 For -C 1-10 Alkyl; the -C 1-10 The alkyl group is optionally substituted by 1, 2 or 3 R c replace; Preferably, the R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted with 1 or 2 R c Substitution, wherein Rc is F, cyclopropyl, cyclobutyl or methyl; More preferably, the R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted with 1 cyclopropyl group; (10) R 5 is H; (11) R a are independently cyano, halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl; preferably cyano, halogen, methyl or cyclopropyl; for example cyano, F or methyl; (12) R b are independently halogen, preferably F; (13) R c are independently halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl; preferably halogen, methyl, cyclopropyl or cyclobutyl; for example cyclopropyl; (14) The n is 0.

5. The use according to claim 1 or 2, It is characterized in that The compound represented by formula I satisfies one or more of the following conditions: (1) R 1 for Best More preferred (2) R 4 for Preferably More preferred (3) for Best (4) for Best More preferred 6. The use according to claim 1 or 2, It is characterized in that The compound represented by formula I satisfies one of the following conditions: (1) The compound represented by formula I is a compound represented by formula IA, Wherein, * represents a chiral carbon atom of S configuration or R configuration; Preferably, Formula IA is Formula IA-1 or Formula IA-2: (2) The compound of formula I is a compound of formula I-1, a compound of formula I-2 or a compound of formula I-3, preferably a compound of formula I-2 or a compound of formula I-3; more preferably a compound of formula I-3: Preferably, the compound represented by formula I-1 is a compound represented by formula I-1-A and / or a compound represented by formula I-1-B. The compound represented by formula I-2 is a compound represented by formula I-2-A and / or a compound represented by formula I-2-B, The compound represented by formula I-3 is a compound represented by formula I-3-A and / or a compound represented by formula I-3-B, 7. The use according to claim 1 or 2, It is characterized in that The compound represented by formula I is selected from any of the following schemes: Scenario 1: The X 1 is N; The X 2 NR X2 , CR X3 or CR X4 R X5 ; The R X2 , the R X3 , the R X4 and the R X5 are independently -H, -C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl; The R 3 is -H or oxo; Said is a single bond or a double bond; The R 1 and the R 2 are independently H, -C(O)R 11 、-C(O)OR 12 、-S(O) 2 R 13 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S", the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the "5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" and the "3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S" are independently optionally replaced by 1, 2 or 3 R a replace; The R 11 , the R 12 and the R 13 Independently -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 aryl, "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" or "a 3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group, the "5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" and the "3-10 membered heterocycloalkyl group having 1, 2 or 3 heteroatoms selected from N, O and S" are independently optionally replaced by 1, 2 or 3 R b replace; The R 4 and the R 5 are independently H, -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R c replace; The R a are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl; The R b are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl; The R c are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-6 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl; The R 6 is cyano; said R 7 is H; said R 8 is halogen; said n is 0 or 1; Scenario 2: The X 1 for CH; The X 2 NR X2 , CR X3 or CR X4 R X5 ; The R X2 , the R X3 , the R X4 and the R X5 are independently -H, -C 1-6 Alkyl, or -C substituted by 1, 2 or 3 halogens 1-6 alkyl; The R 3 is -H or oxo; Said is a single bond or a double bond; The R 1 and the R 2 are independently H, -C(O)R 11 or "a 5-10 membered heteroaryl group wherein the heteroatom is selected from 1, 2 or 3 of N, O and S, and the number of the heteroatom is 1, 2 or 3"; The R 11 For 1, 2 or 3 R b Replaced-C 1-10 alkyl; The R 4 and the R 5 are independently H, -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R c replace; The R b are independently halogen; The R c are independently cyano, halogen, -C 1-10 Alkyl, -OC 1-10 Alkyl, -C 3-10 Cycloalkyl, -OC 3-6 Cycloalkyl or -C 6-10 Aryl; The R 6 is cyano; said R 7 is H; said R 8 is halogen; said n is 0 or 1; Scenario 3: The X 1 is N; The X 2 NR X2 or CR X3 ; The R X2 , the R X3 , the R X4 and the R X5 are independently -H, -C 1-6 Alkyl or -C substituted by 1, 2 or 3 halogens 1-6 alkyl; The R 3 is -H or oxo; Said is a single bond or a double bond; The R 1 H, -C(O)R 11 , -C 1-10 Alkyl, -C 3-10 Cycloalkyl, -C 6-10 Aryl or "a 5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S"; the -C 1-10 Alkyl, the -C 3-10 Cycloalkyl, the -C 6-10 The aryl group and the "5-10 membered heteroaryl group having 1, 2 or 3 heteroatoms selected from N, O and S" are independently optionally replaced by 1, 2 or 3 R a replace; The R 2 is H; The R 11 For -C 1-10 Alkyl, -C 3-10 Cycloalkyl or -C 6-10 Aryl; said -C 1-10 Alkyl, the -C 3-10 Cycloalkyl and the -C 6-10 The aryl group is optionally substituted by 1, 2 or 3 R b replace; The R 4 For -C 1-10 Alkyl; the -C 1-10 The alkyl group is optionally substituted by 1, 2 or 3 R c replace; The R 5 is H; The R a are independently cyano, halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl; The R b are independently halogen; The R c are independently cyano, halogen, -C 1-10 Alkyl or -C 3-6 Cycloalkyl; The R 6 is cyano; said R 7 is H; said R 8 is a halogen; said n is 0; Solution 4: The X 1 CH, X 2 for CH; The R 3 is H; Said is a double bond; The R 1 -C(O)R 11 or "a 5-10 membered heteroaryl group wherein the heteroatom is selected from 1, 2 or 3 of N, O and S, and the number of the heteroatom is 1, 2 or 3"; The R 2 is H; The R 11 For 1, 2 or 3 R b Replaced-C 1-10 alkyl; The R 4 For -C 1-10 Alkyl; the C 1-10 The alkyl group is optionally substituted by 1, 2 or 3 R c replace; The R 5 is H; The R b are independently halogen; The R c are independently halogen; The R 6 is cyano; said R 7 is H; said R 8 is a halogen; said n is 0; Solution 5: Said for The R 3 is -H or oxo; The R 1 -C(O)R 11 , -C 1-4 Alkyl, -C 3-6 Cycloalkyl, phenyl or "a 5-6 membered heteroaryl group having 1 or 2 nitrogen atoms as heteroatoms"; the -C 1-4 Alkyl, the -C 3-6 The cycloalkyl group, the phenyl group and the "5-6 membered heteroaryl group having 1 or 2 heteroatoms as nitrogen" are independently optionally substituted by 1, 2 or 3 R a replace; The R 11 For-CF 3 , cyclopropyl or phenyl; The R 2 is H; The R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted by 1, 2 or 3 R c replace; The R 5 is H; The R 6 is cyano; said R 7 is H; said R 8 is a halogen; said n is 0; Solution 6: Said for The R 3 is -H or oxo; The R 1 -C(O)R 11 , phenyl or "a 5-6 membered heteroaryl group wherein the heteroatom is nitrogen and the number is 1 or 2"; the phenyl group and the "a 5-6 membered heteroaryl group wherein the heteroatom is nitrogen and the number is 1 or 2" are independently optionally replaced by 1, 2 or 3 R a replace; The R 11 For -C 3-6 Cycloalkyl; The R 2 is H; The R 4 For -C 1-4 Alkyl; the -C 1-4 The alkyl group is optionally substituted with 1 cyclopropyl group; The R 5 is H; The R 6 is cyano; said R 7 is H; said R 8 is halogen; and n is 0.

8. The use according to claim 1 or 2, It is characterized in that The compound as shown in formula I is selected from any of the following compounds: Preferably, the compound as shown in formula I is selected from any of the following compounds:

9. A compound, which is a compound as shown in formula II or a compound as shown in formula III, In Formula II, X 1 , X 2 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 The definition as in any one of claims 1 to 8; R 9 for In formula III, or R X2 , R X3 , R 1 , R 2 , R 3 , R 4 and R 5 The definition as described in any one of claims 1 to 8.

10. The use according to claim 9, It is characterized in that The compound shown in formula II is selected from any of the following compounds: The compound shown in formula III is selected from any of the following compounds:

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