Amide compound as well as preparation method, pharmaceutical composition and application thereof

Abstract

The invention provides a compound containing a general formula I as well as a preparation method, a pharmaceutical composition and application thereof. Specifically, the invention provides a compound with a structure as shown in a general formula I, and a raceme, an R-isomer, an S-isomer, pharmaceutically acceptable salt or a mixture thereof. The compound has good inhibitory activity on the PL protease, can be used for treating, preventing and relieving diseases related to the PL protease, and particularly can be used for treating viral diseases with the PL protease, such as viral diseases caused by SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV-OC43 and the like.

Description

Technical Field

[0001] This invention relates to the fields of medicinal chemistry and pharmacotherapeutic science, specifically to a class of compounds containing general formula I as inhibitors of coronavirus papain, methods for their preparation, pharmaceutical compositions containing such compounds, and as inhibitors of papain (also known as PL). pro Inhibitors of protease (PL) or PL protease, especially for the treatment of PL protease. pro Viral diseases, such as those caused by SARS-CoV-2, SARS-CoV, MERS-CoV, and HCoV-OC43. Background Technology

[0002] Coronaviruses (Cov) belong to the order Nidovirales, family Coronaviridae, and genus Coronavirus. They are medium-sized, spherical, enveloped, single-sense positive-sense RNA viruses, approximately 80–160 nm in diameter, with a genome length of about 30 kb. They are the largest known RNA viruses, named for the crown-like appearance of the viral particles, which are enveloped by spike proteins on their envelope, under an electron microscope. Coronaviruses are widely distributed in natural hosts, including humans, mammals, and birds. Infection with coronaviruses typically causes symptoms of respiratory, digestive, and nervous system diseases. The International Committee on Taxonomy of Viruses classifies coronaviruses into four groups: α, β, γ, and δ. Currently, the seven coronaviruses that infect humans mainly include human coronavirus NL63 (HCoV-NL63) and human coronavirus 229E (HCoV-229E) distributed in the α group, and human coronavirus OC43 (HCoV-OC43) and human coronavirus HKU1 (HCoV-HKU1) distributed in the β group, as well as SARS-CoV, MERS-CoV, and SARS-CoV-2. Animal coronaviruses mainly include porcine epidemic diarrhea virus (PEDV), canine coronavirus (CCoV), feline infectious peritonitis virus (FIPV), and mouse hepatitis virus (MHV). Human coronaviruses mainly cause respiratory symptoms. HCoV-229E and HCoV-OC43 usually cause the common cold in winter and spring, while HCoV-NL63 causes more severe clinical symptoms of cough in children, and HCoV-HKU1 mainly causes bronchitis and pneumonia. Compared to the four viruses mentioned above, SARS-CoV, MERS-CoV, and SARS-CoV-2 cause more severe respiratory symptoms and even lead to death. The case fatality rates for the former two are 9.6% and 35.7%, respectively, while the novel coronavirus infection caused by SARS-CoV-2 has spread globally, severely impacting human life. In addition to respiratory symptoms, coronaviruses can also cause neurological symptoms. HCoV-229E, HCoV-OC43, and SARS-CoV have all been detected in cerebrospinal fluid. Patients infected with MERS-CoV and SARS-CoV-2 exhibit certain neurological symptoms, while HCoV-OC43 has been reported to cause fatal encephalitis cases.

[0003] Coronaviruses share similarities in their genome composition and expression. Their open reading frames (ORFs) encode 27 proteins. The genome near the 3′ end encodes viral structural and accessory proteins, including the spike protein (S), nucleocapsid protein (N), membrane protein (M), and envelope protein (E). The S protein primarily mediates viral invasion by binding to host cell receptors; the N protein encapsulates the viral genome to form a nucleoprotein complex; and the M and E proteins are mainly involved in viral replication, assembly, and budding. The genome at the 5′ end contains two large overlapping ORFs: ORF1a and ORF1b, which encode replicase polyprotein 1ab (pp1ab) and replicase polyprotein 1a (pp1a), respectively. These two replicase polyproteins are cleaved into non-structural proteins (NSPS) by two proteolytic enzymes, which participate in the viral replication and transcription process.

[0004] PL pro The structural domain is the most structurally complete part of all coronaviruses, and its amino acid sequence is highly conserved across all coronavirus genera. PL pro Located in NSP3, its main function is to interact with the main protease (M). pro Together, they are responsible for cleaving viral precursor proteins, promoting the formation of mature viral proteins. They cleave the viral polymers pp1a and pp1ab at three sites, producing NSP1, NSP2, and NSP3. pro It contains an N-terminal ubiquitin-like domain (Ubl) and a catalytic core domain. The catalytic core domain comprises three subdomains: an α-helical thumb domain, a palm domain, and a β-sheet zinc finger domain, resembling an open right hand. The boundary between the thumb and palm domains is PL. pro The catalytic active site is composed of a Cys-His-Asp catalytic triplet. The zinc finger domain includes a Zn group tetrahedral coordinated by four cysteine ​​residues. 2+ To maintain PL pro Structural integrity and catalytic activity are crucial. The substrate binding site recognizes the conserved sequence LXGG↓X (the amino acid residues of the substrate are numbered P4-P3-P2-P1↓P1', and the downward arrows indicate cleavage sites). PL proSubsites S1-S4 provide binding sites for substrates P1-P4, respectively. Near the catalytic site is a flexible loop consisting of six amino acids (residues 267-272), called blocking loop 2 (BL2), which plays an important role in controlling substrate entry into the active site.

[0005] Research has found that PL pro It not only plays a crucial role in coronavirus replication but also in its ability to evade host immunity and induce additional inflammation in the host. Targeting PL pro The drug may have both antiviral and anti-inflammatory effects, and holds promise for overcoming M... pro The limitations of RdRP inhibitors in efficacy may provide a scientific approach for better protecting humans from the threat of coronaviruses, but currently PL pro Research on inhibitors has progressed slowly. Currently, only one compound, HL-21, is in Phase I clinical trials for this target, while others are still in the discovery stage. There is a lack of inhibitors with novel structures and good activity; therefore, the development of PL... pro Small molecule inhibitors are particularly important. Summary of the Invention

[0006] The purpose of this invention is to provide a small molecule inhibitor of coronavirus PLpro.

[0007] In a first aspect, the present invention provides a compound having the structure shown in general formula I, and its racemic, R-isomer, S-isomer, pharmaceutically acceptable salt, or mixture thereof:

[0008]

[0009] in,

[0010] X is selected from the following group: CR a CR a R b C = O, N, NR a O, S, S = O, SO2;

[0011] Each R a and R b Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl, C 1-6 Alkoxy;

[0012] Y is selected from the following groups: not present, H, NH2, OH, SH. And Y1 is selected from the following groups: NH, O, S;

[0013] The ring is selected from the following group: saturated or partially saturated C 5-7Carbon ring, C 6-10 Aromatic rings, 5-6 membered heteroaromatic rings, 5-7 membered heteroaromatic rings, 6-20 membered heteroaromatic fused rings; wherein the hydrogens on the above-mentioned cyclic groups are optionally substituted by 1, 2, 3 or 4 substituents selected from the group consisting of: halogens, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy groups, C 3-8 cycloalkyl, C 3-8 Halogenated cycloalkyl, cyano, nitro, amino, C 1-6 Amine, hydroxyl, hydroxymethyl, carboxyl, mercapto, C 1-6 alkylsulfonyl, C 6-10 Aryl, 5-12 membered heteroaryl, 3-12 membered heterocyclic;

[0014] Each R 1 R 2 and R 3 Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 alkoxycarbonyl, C 1-6 Halogenated alkoxy groups, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, cyano, C 2-6 Ester group, nitro group, amino group, C 1-6 Amine, hydroxyl, hydroxymethyl, carboxyl, 5-12 membered heteroaryl, 3-12 membered heterocyclic, -(CH2) p CONR 7 R 8 -NR 8 CO(C 1-6 alkyl);

[0015] n and p are each independently 0, 1, 2, 3 or 4;

[0016] m, q, r, s, and t are each independently 0, 1, 2, 3, or 4;

[0017] Each R 4 Independently selected from the following groups: hydrogen, deuterium, halogen, cyano, nitro, amino, C 1-6 Amine, hydroxyl, hydroxymethyl, carboxyl, mercapto, -S(O)2OH, C 1-6 alkylsulfonyl, R c C substituted or unsubstituted 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy groups, C1-6 Alkylamine, C 3-8 cycloalkyl, C 3-8 Halogenated cycloalkyl, C 6-10 Aryl, 3-12 membered heterocyclic groups, -(CH2) p NH(CH2) p COOR 9 NR 10 (CH2) p -;-O(CH2) p R 14 R 14’ (Examples 104 and 105 are double substitutions)

[0018] R c Selected from the following group: C 6-10 Aryl, 5-7 quinone heteroaryl, C 3-7 cycloalkyl groups, 4-10 membered heterocyclic groups;

[0019] The rings are selected from the following groups: none, 3-12 membered carbon rings (including fused rings, bridged rings and spiro rings), 3-12 membered heteromonocyclic rings, 7-20 membered heteropolycyclic rings (including fused rings, bridged rings and spiro rings), and 5-12 membered heteroaromatic rings;

[0020] Each R 5 and each R 6 Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 alkoxycarbonyl, C 1-6 Halogenated alkoxy groups, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, C 3-8 Halogenated cycloalkyl groups, -(CH2) p -Cyano, Nitro, C 2-6 Ester group, amino group, C 1-6 Amine group, hydroxyl group, ketone carbonyl group, C 1-6 Alkyl hydroxyl, carboxyl, mercapto, C 1-6 Alkyl sulfonyl group, -(CH2) p S(O)2R 13 C6-C10 aryl, 5-12 heteroaryl, 3-12 heterocyclic, -C(O)(C 1-6 Hydroxyalkyl), -(CH2) p NR 11 R 11’ -(CH2) p NH(C 1-6 Alkyl group), -(CH2) pNH(CH2) p COOR 12 -(CH2) p COOH;

[0021] R 7 R 8 R 9 R 10 R 11 R 11’ R 12 R 13 R 14 and R 14’ Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 alkoxycarbonyl, C 1-6 Halogenated alkoxy groups, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, cyano, C 2-6 Ester group, nitro group, amino group, C 1-6 Amine, hydroxyl, hydroxymethyl, carboxyl, C 6-10 aryl, 5-12 membered heteroaryl, 3-12 membered heterocyclic; or R 7 R 8 The atoms bonded to it together form 5-10 membered heterocyclic groups;

[0022] Unless otherwise specified, each of the heteroaryl, heteroaromatic ring, heterofused ring, heterocyclic group and heterocycle has 1, 2, 3 or 4 heteroatoms selected from N, O and S on its ring skeleton; the heterocycle or heterocyclic group includes saturated or partially unsaturated cyclic groups;

[0023] The alkyl, alkoxy, alkenyl, alkynyl, cycloalkane, cycloalkyl, heterocyclic, heterocyclic, aryl, and heteroaryl groups are each independently substituted by 1-3 substituents selected from the group consisting of: halogen, C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkoxycarbonyl, C 1-6 Halogenated alkoxy groups, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Cycloalkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, carboxyl, mercapto, C 1-6 alkylsulfonyl, C 6-10 Aryl, 3-12 membered heterocyclic groups;

[0024] The halogen is F, Cl, Br or I.

[0025] In another preferred embodiment, n and p represent the number of alkylene groups, and n and p are each independently 0, 1, 2, 3 or 4.

[0026] In another preferred embodiment, m, q, r, s, and t represent the number of substituents, and m, q, r, s, and t are each independently 0, 1, 2, or 3.

[0027] In a preferred embodiment, the The ring is selected from the following group: saturated or partially saturated C 5-7 Carbon ring, C 6-10 Aromatic rings, 5-6 quintone heterocyclic aromatic rings;

[0028] Preferably, the The ring is selected from the group consisting of: phenyl, furanyl, thiophene, pyrrolyl, thiazolyl, thiadiazolyl, diazolyl, imidazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1,2,4,5-tetraazinyl.

[0029] In a preferred embodiment, the The rings are selected from the following group: 4-10 saturated carbon rings, 4-10 saturated carbon bridged rings, 4-10 saturated or partially saturated heteromonocyclic rings, 7-10 fused heterocyclic rings, 7-10 fused heterobridged rings, 7-10 fused heterospirocyclic rings, and 5-9 fused heteroaromatic rings.

[0030] Preferably, the The ring is selected from the following group:

[0031]

[0032] Where i is 0, 1, 2, 3 or 4.

[0033] In another preferred embodiment, the Selected from the following group:

[0034]

[0035] Where i is 0, 1, 2, 3 or 4.

[0036] In a preferred embodiment, the Selected from the following group:

[0037]

[0038] Where i is 0, 1, 2, 3 or 4.

[0039] In another preferred embodiment, each R 5 and each R 6Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 alkoxycarbonyl, C 1-4 Halogenated alkoxy groups, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-5 cycloalkyl, C 3-5 Halogenated cycloalkyl groups, -(CH2) p -Cyano, Nitro, C 2-4 Ester group, amino group, C 1-4 Amine, hydroxyl, ketone carbonyl, hydroxymethyl, carboxyl, mercapto, C 1-4 Alkyl sulfonyl group, -(CH2) p S(O)2OH, phenyl, naphthyl, 5-7 membered heteroaryl, 4-7 membered heterocyclic group, -C(O)(C 1-4 Hydroxyalkyl), -(CH2) p N(C 1-4 Alkyl group 2, -(CH2) p NH(C 1-4 Alkyl group), -(CH2) p NH(CH2) p COOR 12 -(CH2) p COOH.

[0040] In another preferred embodiment, each R a and R b Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-4 Alkyl, C 1-4 Alkyl group.

[0041] In another preferred embodiment, R c Selected from the following group: C 6-10 Aryl, 5-7 quinone heteroaryl, C 3-6 Cycloalkyl, 4-6 membered heterocyclic groups.

[0042] In another preferred embodiment, each R 1 R 2 and R 3 Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 alkoxycarbonyl, C 1-4 Halogenated alkoxy groups, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-5 cycloalkyl, cyano, C 2-4Ester group, nitro group, amino group, C 1-4 Amine, hydroxyl, hydroxymethyl, carboxyl, 5-7 membered heteroaryl, 4-7 membered heterocyclic, -(CH2) p CONR 7 R 8 -NR 8 CO(C 1-4 alkyl).

[0043] In another preferred embodiment, each R 1 Independently selected from the following groups: hydrogen, fluorine, chlorine, bromine, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkyl group, cyano group.

[0044] In another preferred embodiment, R2 is selected from the group consisting of: hydrogen, deuterium, F, Cl, Br, and C. 1-4 Alkyl, C 1-4 Fluoroalkyl, C 1-4 Alkoxy, C 1-4 alkoxycarbonyl, C 1-4 Halogenated alkoxy groups, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-5 cycloalkyl, cyano, C 2-4 Ester group, nitro group, amino group, C 1-4 Amino, hydroxy, hydroxymethyl, carboxyl, 5-7 membered heteroaryl, 4-7 membered heterocyclic, -CONR 7 R 8 -NR 8 CO(C 1-4 alkyl).

[0045] In another preferred embodiment, the R 3 Selected from the following group: hydrogen, C 1-4 Alkyl, C 1-4 Fluoroalkyl, C 3-5 cycloalkyl;

[0046] Preferably, the R 3 Selected from the following group: hydrogen, methyl, ethyl, trifluoromethyl, cyclopropyl, cyclobutyl, cyclopentyl.

[0047] In another preferred embodiment, the R 4 For unreplaced C 1-4 Alkyl; preferably, the R 4 It is a methyl group.

[0048] In another preferred embodiment, the R 7 R 8 R 9 R 10 R 11 R11’ R 12 and R 13 Each element is independently selected from the following groups: hydrogen, deuterium, halogens, and carbon. 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 alkoxycarbonyl, C 1-4 Halogenated alkoxy groups, C 2-4 alkenyl, C 2-4 alkynyl group, C 3-5 cycloalkyl, cyano, C 2-4 Ester group, nitro group, amino group, C 1-4 Amino, hydroxy, hydroxymethyl, carboxyl, phenyl, naphthyl, 5-7 membered heteroaryl, 4-7 membered heterocyclic, or R 7 R 8 The atoms attached to it together form 5-7 membered heterocyclic groups.

[0049] In a preferred embodiment, the structure of the compound is shown in general formula II:

[0050]

[0051] Among them, s, t, R 1 R 2 R 5 and R 6 The definition is as described above.

[0052] In a preferred embodiment, the compound is one of the compounds shown in the table below:

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074] A second aspect of the present invention provides a pharmaceutical composition comprising: one or more of a compound of formula I as described in the first aspect of the present invention, a pharmaceutically acceptable salt thereof, a racemic mixture, an R-isomer, an S-isomer, or a mixture thereof, and one or more pharmaceutically acceptable carriers, excipients, adjuvants, excipients, and / or diluents.

[0075] A third aspect of the invention provides the use of a compound of formula I as described in the first aspect of the invention, a pharmaceutically acceptable salt thereof, a racemic mixture, an R-isomer, an S-isomer, or a mixture thereof, or a pharmaceutical composition as described in the second aspect of the invention, for the preparation of a pharmaceutical composition for the treatment or prevention of diseases associated with PL protease activity.

[0076] In a preferred embodiment, the disease is caused by a virus containing the PL protease; preferably, the virus is selected from the group consisting of SARS-CoV-2, SARS-CoV, MERS-CoV, and HCoV-OC43, or combinations thereof.

[0077] It should be understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here. Detailed Implementation

[0078] Through long-term and in-depth research, the inventors have designed and synthesized a novel class of quinolinone amide compounds. These compounds exhibit excellent inhibitory activity against PL protease, and therefore can be used to treat, prevent, and alleviate diseases related to PL protease, particularly viral diseases involving PL protease, such as those caused by SARS-CoV-2, SARS-CoV, MERS-CoV, and HCoV-OC43. Based on these findings, the inventors have completed this invention.

[0079] the term

[0080] As used herein, the terms “containing” or “including (comprise)” can be open-ended, semi-closed, or closed-ended. In other words, the terms also include “consistently made of” or “made of”.

[0081] As used herein, the term "alkyl" refers to a monovalent, straight-chain or branched saturated hydrocarbon group consisting of carbon and hydrogen atoms, for example, "C 1-6 "Alkyl" means an alkyl group having 1 to 6 carbon atoms (e.g., 1, 2, 3, 4, 5, or 6). Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, or tert-butyl. In this application, alkyl is also intended to include deuterated alkyl groups, examples of which include, but are not limited to, CD3, CD2CD3, and CD2CD2CD3.

[0082] As used herein, the term "alkylene" refers to the group obtained by removing a hydrogen atom from an alkyl group as described above, such as methylene (-CH2-), ethylene (-CH2CH2-), etc.

[0083] As used herein, the term "alkenyl" refers to a straight-chain or branched unsaturated hydrocarbon group consisting of carbon and hydrogen atoms and having at least one double bond, such as "C 2-6 "Alkenyl" refers to an alkenyl group having 2 to 6 (e.g., 2, 3, 4, 5, or 6) carbon atoms. Examples of alkenyl groups include, but are not limited to: vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, and 1,3-butadienyl.

[0084] As used herein, the term "alkynyl" refers to a straight-chain or branched unsaturated hydrocarbon group consisting of carbon and hydrogen atoms and having at least one triple bond. For example, "C 2-6 "Alynyl" refers to an alkynyl group having 2 to 6 (e.g., 2, 3, 4, 5, or 6) carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, and 1-butynyl.

[0085] As used herein, the term "carbocyclic" or "carbocyclic group" refers to a saturated or partially saturated carbocyclic group composed of carbon and hydrogen atoms, such as a monocyclic, bicyclic, or tricyclic structure, wherein the ring structure as a whole is not aromatic but may contain one or more unsaturated structures. The ring may be further substituted by one or more substituents. When two or more rings are present in the carbocyclic group, the rings may further form fused rings, bridged rings, spirocyclic rings, or any combination thereof.

[0086] As used herein, the term "carbon bridged ring" refers to a saturated or partially saturated bridged ring group composed of carbon and hydrogen atoms.

[0087] As used herein, the term "cycloalkyl" refers to a monovalent saturated carbocyclic group consisting of carbon and hydrogen atoms, such as "C". 3-8 "Cycloalkyl" refers to a cycloalkyl group containing 3 to 8 (e.g., 3, 4, 5, 6, 7, or 8) carbon atoms, preferably C14. 3-6 Cycloalkyl groups. Cycloalkyl groups can be monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or similar groups, or bicyclic, such as fused rings, bridged rings or spirocyclic rings.

[0088] As used herein, the term "alkoxy" refers to the formula -OR z or -R z '-OR z Group, wherein R z R is an alkyl group as defined in this article. z ' is an alkylene group. Examples of alkoxy groups include, but are not limited to: methoxy, ethoxy, isopropoxy, tert-butoxy, -CH2O-CH3, -CH2CH2-O-CH3, -CH2-O-CH2CH3, etc.

[0089] As used herein, the term "hydroxyalkyl" refers to an alkyl group containing one or more hydroxyl groups, and the definition of an alkyl group is as described above. Examples of hydroxyalkyl groups include, but are not limited to, -CH2OH, -CH2CH2OH, -CH2CH2CH2OH, etc.

[0090] As used herein, "halogen" refers to halogens and their isotopes, including but not limited to F, 18 F, 32 Cl, Br, I.

[0091] As used in this article, the term "nitro" refers to -NO2.

[0092] As used in this article, the term "cyano" refers to -CN.

[0093] As used in this article, the term "amino" refers to -NH2.

[0094] As used in this article, the term "thiol" refers to -SH.

[0095] As used in this article, the term "carboxyl group" refers to -COOH.

[0096] As used in this article, the terms “oxo” and “ketone carbonyl” refer to the =O group.

[0097] As used herein, the term "alkoxycarbonyl" refers to -C(O)-alkoxy, and examples of alkoxycarbonyl include, but are not limited to, -COCH2OCH3, -COCH2OCH2CH3, -COCH2OCH(CH3)2, etc.

[0098] As used herein, the term "ester group" refers to -COOR y , where R y It can be independently selected from the group consisting of: hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocyclic groups. Examples of ester groups include, but are not limited to: -COOCH3, -COOCH2CH3, -COOCH2CH2CH3, -COOCH2CH(CH3)2, etc.

[0099] As used herein, the term "amine" refers to -NR. w R w ', where R w and R w It can be independently selected from the group consisting of: hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocyclic. R w and R w 'They can be the same or different, but R' w and R w 'Not both hydrogen. Examples of amino groups include, but are not limited to: -NHCH3, -SO2N(CH3)2, etc.

[0100] As used herein, the term "alkylamine" refers to -NR w R w ', where R w and R w It can be independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl groups. R w and R w 'They can be the same or different, but R' w and R w 'Not both hydrogen. Examples of amino groups include, but are not limited to: -NHCH3, -SO2N(CH3)2, etc.

[0101] As used herein, the term "haloalkyl" refers to a group obtained by substituting one or more hydrogen atoms in an alkyl group as described above with the same or different halogens. Wherein, "C..." 1-6 "Halogenated alkyl" is preferably C 1-4 Haloalkyl groups, examples of which include, but are not limited to: -CH2Cl, -CH2CF3, -CH2CCl3, perfluoroalkyl groups (e.g., -CF3-, -CF2CF3), etc.

[0102] As used herein, the term "halocycloalkyl" refers to a group obtained by substituting one or more hydrogens in a cycloalkyl group as described above with the same or different halogens.

[0103] As used herein, the term "haloalkoxy" refers to a group obtained by substituting one or more hydrogen atoms in an alkoxy group as described above with the same or different halogens. Examples of haloalkoxy groups include, but are not limited to, trifluoromethoxy, difluoromethoxy, 2,2,2-trifluoroethoxy, etc.

[0104] As used herein, the term "alkylsulfonyl" refers to the formula -S(O)2-alkyl. The sulfonyl group is preferably -S(O)2-(C 1-6 Alkyl groups, such as -S(O)2-CH3, -S(O)2-CH2CH3, etc.

[0105] As used herein, the term "heterocyclic" or "heterocyclic group" refers to a fully or partially saturated monocyclic, bicyclic, or polycyclic cyclic group on a ring backbone containing one or more heteroatoms selected from N, S, or O. For example, "4-7 membered heterocyclic group" refers to a group having 4-7 (e.g., 4, 5, 6, or 7) ring members. The nitrogen or sulfur atom may be oxidized, or the nitrogen atom may be quaternized. The heterocyclic group can be attached to any heteroatom or carbon residue in a ring or ring system molecule. Monocyclic heterocyclic groups include, but are not limited to: azacyclic butyl, pyrrolyl, oxacyclic butyl, pyrazolinyl, imidazolinyl, imidazoalkyl, oxazolinyl, isoxazolinyl, thiazoalkyl, isothiazolinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperidinyl, 2-oxopiperidinyl, 2-oxopiperylyl, hexahydroachenginyl, 4-piperidinoneyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxane, and tetrahydro-1,1-dioxothiophene, etc. Polycyclic heterocyclic groups include, but are not limited to, spirocyclic, fused-ring, and bridged-ring heterocyclic groups; wherein the spirocyclic, fused-ring, and bridged-ring heterocyclic groups involved are optionally connected to other groups by single bonds, or further cyclically linked to other cycloalkyl, heterocyclic, aryl, and heteroaryl groups by any two or more atoms on the ring.

[0106] As used herein, the term "heterocyclic monocyclic" refers to a monocyclic group on a ring backbone that is fully or partially saturated with one or more heteroatoms selected from N, S, or O heteroatoms.

[0107] As used herein, the term "heterocyclic fused ring" refers to a ring skeleton containing one or more fully or partially saturated fused ring groups selected from N, S, or O heteroatoms.

[0108] As used herein, the term "heterospirocyclic" refers to a ring skeleton containing one or more fully or partially saturated spirocyclic groups selected from N, S, or O heteroatoms.

[0109] As used herein, the term "heterobridged ring" refers to a ring skeleton containing one or more fully or partially saturated bridging ring groups selected from N, S, or O heteroatoms.

[0110] As used herein, the term "aromatic ring" or "aryl" refers to an aromatic cyclic hydrocarbon group (including monocyclic, bicyclic, or polycyclic groups), such as "C 6-12 "Aryl" refers to an aromatic cyclic hydrocarbon group having 6-12 (6, 7, 8, 9, 10, 11, or 12) ring carbon atoms. It contains two or more aromatic rings (such as bicyclic rings), and the aromatic rings of the aryl group can be linked by single bonds (such as biphenyl) or fused (such as naphthalene, anthracene, etc.). Examples of aryl groups (especially monocyclic and bicyclic groups) include, but are not limited to, phenyl, biphenyl, or naphthyl. Aryl groups can be fused with heterocyclic groups through single bonds or any two adjacent ring carbon atoms, for example: benzotetrahydrofuranyl, benzotetrahydropyranyl, benzodioxane, etc. wait.

[0111] As used herein, the term “heteroaromatic ring” or “heteroaryl” refers to an aromatic cyclic group (including monocyclic, bicyclic, or polycyclic groups) whose ring skeleton contains 1, 2, 3, or 4 heteroatoms selected from N, S, or O. For example, “5-12-membered heteroaryl” refers to a monocyclic, bicyclic, or tricyclic group having 5 to 12 (5, 6, 7, 8, 9, 10, 11, or 12) ring atoms. Examples of heteroaryl groups include, but are not limited to: imidazole, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyrazinyl, thiophene, furanyl, pyranyl, pyridinyl, pyrroleyl, pyrazolyl, pyrimidinyl, quinolinyl, isoquinolinyl, benzofuranyl, benzothiophene, benzothiaranyl, benzimidazole, benzooxazolyl, benzooxadiazolyl, benzothiazolyl, benzopyranyl, indoleyl, isoindoleyl, triazolyl, triazinyl, quinoxolinyl, purine, quinazolinyl, quinazinyl, naphthidyl, pteridinyl, carbazoleyl, and azazolyl. basalt, diazoxide acridine group, etc.

[0112] As used in this article, the term "multi-substitution" refers to a substance that includes two or more substitutions.

[0113] When a group loses one hydrogen atom, it becomes a subunit of the corresponding group, and it is a divalent group. For example, an alkyl group loses one hydrogen atom to become an alkylene group (e.g., methylene, ethylene, propylene, isopropylene). ), butylide (such as) ), pentylene (e.g.) ), hexyl (such as) ), subheptagen (such as ) etc.; cycloalkyl corresponds to cyclohexane (e.g.: (etc.); heterocyclic groups correspond to subheterocyclic groups (e.g.: Alkoxy groups correspond to alkoxy groups (e.g., -CH2O-, -CH2CH2O-, -OCH2CH2CH2-), and heteroalkyl groups correspond to heteroalkyl groups (e.g., -CH2-O-CH2CH2-, -CH2-O-(CH2)2CH2-, -CH2CH2-O-CH2CH2-, -CH2-O-CH2CH2CH2-, -CH2-S-CH2CH2-, -CH2-S-(CH2)2CH2-, -CH2CH2-S-CH2CH2-, -CH2-S-CH2CH2CH2-, -CH2-NH-CH2CH2-, -CH2-NH-(CH2)2CH2-, -CH2CH2-NH-CH2CH2-, -CH2-NH-CH2CH2CH2-, etc.).

[0114] In this invention, the alkyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups, unless otherwise specified, include substituted alkyl, cycloalkyl, heterocyclic, aryl, and heteroaryl groups. The substituents include, but are not limited to, halogen, hydroxyl, cyano, acyl, sulfonyl, ester, sulfinyl, alkyl, cycloalkyl, heterocyclic, aryl, heteroaryl, acyl, and ester groups.

[0115] As used herein, the term "substitution" refers to the replacement of one or more hydrogen atoms on a particular group by a particular substituent. The particular substituent is either the substituent described accordingly above or the substituent appearing in the various embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a particular group at any substituted site of that group, and the substituents may be the same or different at each position. Those skilled in the art will understand that the combinations of substituents contemplated in this invention are those that are stable or chemically feasible.

[0116] Unless otherwise specified, the groups described in this invention may be substituted with substituents selected from the group consisting of: D, halogen, cyano, nitro, hydroxyl, amino, C. 1-6 Alkyl, C2-C6 alkenyl, C2-C6 ynyl, C1-6 Alkoxy, 3-12 membered heterocyclic group, C3-C 12 cycloalkyl, 5-10 heteroaryl and C6-C 10 Aryl.

[0117] In this document, “optionally” means that the event or condition described below may, but is not required to, occur, and the description includes both the occurrence of the event or condition and the non-occurrence of the event or condition.

[0118] In this article, the term "multiple" refers to 2, 3, 4, 5, or a positive integer greater than 5.

[0119] Active ingredients

[0120] As used herein, “compound of the present invention” means a compound of formula (I), and also includes its stereoisomers, its optical isomers, its pharmaceutically acceptable salts, its crystal forms, its isotopic derivatives, its prodrugs, its metabolites, its solvates or hydrates thereof.

[0121] Unless otherwise specified, the structural formulas described in this invention are intended to include all stereoisomers (such as cis-trans isomers, enantiomers, diastereomers, and conformational isomers): R and S configurations containing an asymmetric center, (Z) and (E) isomers of double bonds, cis-trans isomers of cycloalkanes, etc. Therefore, any single stereochemical isomer of the compounds of this invention, or a mixture of its enantiomers, diastereomers, or conformational isomers, is within the scope of this invention.

[0122] The compounds of this invention may contain cis-trans isomers, one or more chiral carbon atoms, and thus can produce cis-trans isomers, chiral isomers, enantiomers, diastereomers, and other combinations of stereoisomers. Cis-trans isomerism refers to the diastereomeric phenomenon in which different groups in a compound molecule are arranged differently in space due to a restrictive factor that limits free rotation. This restrictive factor is generally caused by non-rotating functional groups in the structure of organic compounds, such as C=C double bonds, C=N double bonds, C=S double bonds, N=N double bonds, heterocycles, or cycloalkanes. Organic molecules containing such isomers, such as alkenes, azo compounds, and alicyclic hydrocarbons, are considered cis-trans isomers. Cis refers to the same ligands being in adjacent positions, generally denoted by "cis" or "cis-"; trans refers to the same ligands being in diagonal positions, generally denoted by "trans" or "trans-". Each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry. This invention aims to include all possible isomers, their racemic and optically pure forms. The compounds of this invention can be prepared using racemic, cis-trans, chiral, diastereomer, or enantiomers as starting materials or intermediates. Optically active isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0123] Conventional techniques for preparing / separating individual optical isomers (i.e., cis-trans isomers and chiral isomers) include chiral synthesis from suitable cis-trans precursors or optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography.

[0124] To design the synthesis of a specific stereoisomer of the compound of this invention, it can be prepared asymmetrically or derivatized with a chiral auxiliary. The resulting stereo mixture is then separated, and the chiral auxiliary is removed to obtain pure cis-trans monomers, chiral monomers, or mixed stereoisomers. If the molecule contains a cis-trans isomer center, it can be purified by column chromatography (normal-phase silica gel column or reverse-phase high-performance liquid chromatography) to obtain pure cis or trans products. Alternatively, if the molecule contains a basic functional group, such as an amino acid, or an acidic functional group, such as a carboxyl group, it can be formed with a suitable optically active acid or base to form a diastereomeric salt, which is then separated by conventional methods such as separation crystallization or chromatography to obtain pure enantiomers.

[0125] This invention also includes isotopically labeled compounds (i.e., isotopic derivatives), equivalent to the original compounds disclosed herein. However, it is common practice to see one or more atoms replaced by atoms with different atomic weights or mass numbers. Examples of isotopes in the isotopic derivatives of this invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine isotopes, respectively as follows: 2 H, 3 H,13 C 11 C 14 C 15 N、 18 O、 17 O、 31 P, 32 P, 35 S, 18 F and 36 Cl. Isotope derivatives of the compounds of this invention are all within the scope of protection of this invention. In this document, 3 H-labeled compounds and 14 C-labeled compounds are useful in tissue distribution experiments of drugs and substrates. Tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) Labeled compounds are relatively easy to prepare and detect, making them the preferred choice among isotopes. Furthermore, heavier isotope substitutions, such as deuterium, are also possible. 2 H, due to its excellent metabolic stability, offers advantages in certain therapies, such as increasing half-life or reducing dosage in vivo, and therefore may be preferred in some cases. Isotopically labeled compounds can be prepared using general methods, by replacing the non-isotopic reagent with an readily available isotopically labeled reagent, according to the scheme disclosed in the examples.

[0126] As used herein, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0127] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the bioavailability of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochlorides, hydrobroms, sulfates, nitrates, and phosphates; organic acid salts include, but are not limited to, formates, acetates, 2,2-dichloroacetate, trifluoroacetate, propionates, hexanoates, octanoates, decanoates, undecenoates, glycolates, gluconates, lactates, sebates, adipates, glutarate, malonates, oxalates, maleates, succinates, fumarates, tartrates, citrates, palmitates, stearates, oleates, cinnamates, laurates, malates, glutamates, pyroglutamates, aspartate, benzoates, methanesulfonates, benzenesulfonates, p-toluenesulfonates, alginates, ascorbic acid salts, salicylates, 4-aminosalicylic acid salts, and naphthalene disulfonates. These salts can be prepared using methods known in this field.

[0128] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following: primary amines, secondary amines, and tertiary amines; substituted amines, including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

[0129] Metabolites of the compound represented by formula (I) and its pharmaceutically acceptable salts, as well as prodrugs that can be converted in vivo into the compound represented by formula (I) and its pharmaceutically acceptable salts, are also included within the scope of protection of this invention.

[0130] As used herein, the term "solvent" refers to a complex of a compound of formula (I) coordinated with a solvent molecule in a specific ratio.

[0131] As used herein, the term "hydrate" refers to a complex of the compound represented by formula (I) coordinated with water molecules in a specific ratio.

[0132] As described herein, the compounds of the present invention can be substituted with any number of substituents or functional groups to broaden their scope. Generally, the term "substitution" refers to replacing a hydrogen radical with a substituent of a specified structure. When multiple positions in a particular structure are substituted by multiple specific substituents, each position of the substituent can be the same or different. The term "substitution" as used herein includes all permissible organic group substitutions. In a broad sense, permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic organic groups. As described herein, heteroatomic nitrogen may be supplemented with a hydrogen substituent or any permissible organic group described above to complete its valence state. Furthermore, the present invention is not intended to limit permissible substituted organic groups in any way. The present invention considers the combination of substituents and variable groups to be beneficial in the treatment of diseases in the form of stable compounds. The term "stable" here means having a stable compound that, when tested over a sufficiently long period, maintains sufficient integrity of the compound structure, preferably remaining effective for a sufficiently long period, and is used herein for the purposes described above.

[0133] Preparation method of compound I

[0134] The present invention also provides a method for preparing a compound represented by general formula I, the method being carried out according to the following scheme (example):

[0135]

[0136] Step a: Compound 1 is dissolved in a solvent, potassium hydroxide and carbon disulfide are added, and the mixture is heated for 24 hours to obtain compound 2; the solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane, water or a mixture thereof; the heating temperature range is 50-80℃;

[0137] Step b: Compound 2 is dissolved in an organic solvent, and ethyl 2-bromobutyrate and potassium carbonate are added. The mixture is heated and stirred until the reaction is complete to obtain compound 3. The organic solvent is tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane, or a mixture thereof. The heating temperature range is 50–80°C.

[0138] Step c: Compound 3 was dissolved in a mixed solvent of tetrahydrofuran, methanol, and water, followed by the addition of potassium hydroxide, and the mixture was heated under reflux for 1 hour. After the reaction was complete, the organic solvent was removed by rotary evaporation under reduced pressure. The pH was adjusted to acidic with 1M dilute hydrochloric acid, then diluted with water and extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 4.

[0139] Step d: Compound 4 was dissolved in an organic solvent, followed by the addition of HATU, DIPEA, and an amine, and reacted at room temperature for 10 h to obtain compound 5. The organic solvent was tetrahydrofuran, diethyl ether, dimethylformamide, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, dioxane, ethanol, methanol, ethyl acetate, dichloromethane, or a mixture thereof.

[0140] Pharmaceutical Compositions and Administration

[0141] Because the compounds of the present invention have excellent PL protease inhibitory activity, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used for viral diseases caused by PL protease, such as diseases caused by SARS-CoV-2, SARS-CoV, MERS-CoV and HCoV-OC43.

[0142] The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-3000 mg of the compound of the present invention per dose (active dose range 3-30 mg / kg), more preferably, 10-2000 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0143] "Pharmaceutically acceptable carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as... Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0144] There are no particular limitations on the administration of the compounds or pharmaceutical compositions of the present invention. Representative administration methods include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and local administration.

[0145] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0146] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0147] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0148] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0149] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0150] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0151] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0152] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0153] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to the mammal (such as a human) requiring treatment. The dosage administered is the pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 6–600 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the patient's health condition, which are all within the scope of the skill of a skilled physician.

[0154] The main advantages of this invention are:

[0155] This invention provides a novel PL main protease inhibitor. Based on previous compounds, the connection mode of the benzene ring and B ring in the amide side chain is improved, thereby effectively improving the PL main protease inhibitory activity of the compound. It can be used to prepare pharmaceutical compositions for treating diseases caused by viral infections (such as SARS, MERS, 2019n-CoV).

[0156] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

[0157] Example 1 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (1)

[0158]

[0159] 1.1 Preparation of intermediate 1H-imidazolium[4,5-b]pyridine-2-thiol

[0160] 2,3-Diaminopyridine (10 g, 91.6 mmol) was dissolved in 88 mL of ethanol and 18 mL of water, and potassium hydroxide (10.28 g, 183.26 mmol) was added, followed by carbon disulfide (13.95 g, 183.26 mmol). The mixture was refluxed at 80 °C for 24 hours. After the reaction was complete, the solvent was removed by rotary evaporation under reduced pressure to obtain a brown solid. The crude product was directly used in the next reaction without further separation and purification.

[0161] 1.2 Preparation of intermediate 2-((4-fluorobenzyl)thio)-1H-imidazo[4,5-b]pyridine

[0162] 1H-imidazolium[4,5-b]pyridine-2-thiol (5 g, 33.07 mmol) was dissolved in ethanol, and triethylamine (3.35 g, 33.07 mmol) and p-fluorobenzyl bromide (6.25 g, 33.07 mmol) were added. The mixture was refluxed at 60 °C for 1 hour. After the reaction was completed, the mixture was rotary evaporated under reduced pressure, filtered, and the filter cake was washed with anhydrous ethanol (10 mL × 3). The residue was dried to give 6.1 g of white solid, with a two-step yield of 71%.

[0163] 1.3 Preparation of intermediate ethyl 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyrate

[0164] 2-((4-fluorobenzyl)thio)-1H-imidazo[4,5-b]pyridine (2 g, 7.71 mmol) and ethyl 2-bromobutyrate (3.01 g, 15.43 mmol) were dissolved in N,N-dimethylformamide, followed by the addition of potassium carbonate (3.2 g, 23.14 mmol), and the mixture was refluxed at 60 °C for 12 hours. After the reaction was complete, the mixture was diluted with a large amount of water and extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using petroleum ether / ethyl acetate = 1.5 / 1 (v / v) as the eluent to obtain a yellow oily substance in 43% yield.

[0165] 1.4 Preparation of intermediate 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyric acid

[0166] Ethyl 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-1-yl)butyrate (300 mg, 0.8 mmol) was dissolved in 10 mL of tetrahydrofuran, 5 mL of methanol, and 5 mL of water. Potassium hydroxide (225 mg, 4.0 mmol) was then added, and the mixture was refluxed at 60 °C for 1 hour. After the reaction was complete, the organic solvent was removed by rotary evaporation under reduced pressure. The pH was adjusted to acidic with 1 M dilute hydrochloric acid, then diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was directly used in the next reaction without further purification.

[0167] 1.5 Preparation of intermediate 1-methyl-4-(4-methyl-3-nitrophenyl)piperazine

[0168] 4-Bromo-1-methyl-2-nitrobenzene (5 g, 23.14 mmol) was dissolved in 1,4-dioxane, followed by the addition of N-methylpiperazine (3.48 g, 34.72 mmol), bis(dibenzylacetone)palladium (423.88 mg, 0.463 mmol), 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (441.35 mg, 0.926 mmol), and cesium carbonate (15.08 g, 46.29 mmol). The reaction was carried out under nitrogen protection and refluxed at 100 °C for 12 hours. After the reaction was complete, the mixture was filtered, and the filtrate was concentrated by rotary evaporation under reduced pressure to obtain the crude product. Column chromatography was performed using petroleum ether / ethyl acetate (1 / 1, v / v) as eluent to obtain a yellow solid in 78% yield.

[0169] 1.6 Preparation of intermediate 2-methyl-5-(4-methyl-1-piperazinyl)aniline

[0170] The intermediate 1-methyl-4-(4-methyl-3-nitrophenyl)piperazine (4 g, 17 mmol) was dissolved in methanol, and 400 mg of 10% palladium on carbon (55% water) was added. The reaction was carried out at room temperature and atmospheric pressure for 4 hours. After the reaction was completed, the mixture was filtered and concentrated under reduced pressure to obtain the crude product. The crude product was directly used for the next reaction without further separation and purification.

[0171] 1.7 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (1)

[0172] The intermediate 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyric acid (200 mg, 0.58 mmol) was dissolved in dichloromethane. Oxaloyl chloride (166.35 mg, 1.45 mmol) and a catalytic amount of N,N-dimethylformamide were added at 0 °C. After 5 minutes, the mixture was moved to room temperature and reacted for 2.5 hours. Upon completion of the reaction, the mixture was concentrated under reduced pressure to obtain the crude product. The crude product, 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyryl chloride, was directly used in the next reaction without further purification. The intermediate 2-methyl-5-(4-methylpiperazin-1-yl)aniline was dissolved in dichloromethane, and triethylamine (222.50 mg, 2.20 mmol) was added. Crude 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyryl chloride was added at 0 °C, and the mixture was allowed to react overnight at room temperature after 5 minutes. After the reaction was complete, the crude product was concentrated under reduced pressure and separated by column chromatography using dichloromethane / methanol = 20 / 1 (v / v) as eluent to obtain 223 mg of off-white solid, yield 50%. LRMS(ESI) m / z 611 (M+). 1 H NMR (600MHz, DMSO-d6) δ10.06(s,1H),8.46(s,1H),8.27(s,1H),7.54(s,2H),7.13-7.06(m,2H),7.05(d,J=8.3Hz,1H),6.93(s,1H),6.72(d,J=8 .2Hz,1H),6.09(s,1H),4.55(s,2H),3.09(s,4H),2.61(s,3H),2.43(s, 2H), 2.32 (s, 3H), 2.09 (s, 3H), 2.05-1.91 (m, 1H), 0.86 (t, J = 6.1Hz, 3H).

[0173] Example 2 Preparation of 2-(2-((4-fluorobenzyl)thio)-6-(trifluoromethyl)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (2)

[0174] 2,3-Diaminopyridine was replaced with 5-trifluoromethylpyridine-2,3-diamine, and the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 2. LRMS(ESI) m / z 602 (M+).

[0175] Example 3 Preparation of N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)-2-(2-((4-(pyrrolidine-1-carbonyl)benzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (3)

[0176] Replacing p-fluorobenzyl bromide with (4-(bromomethyl)phenyl)(pyrrolidone-1-yl)methyl ketone, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 3. LRMS(ESI) m / z 612 (M+).

[0177] Example 4 Preparation of 2-(2-((4-acetamidobenzyl)thio)-6-bromo-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (4)

[0178] Replacing p-fluorobenzyl bromide with N-(4-(bromomethyl)phenyl)acetamide, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 4. LRMS(ESI) m / z 650 (M+). 1 H NMR (600MHz, DMSO) δ10.02(d,J=13.8Hz,1H),8.46(s,1H),8.24(d,J=40.6Hz,1H),7.56-7.49( m,2H),7.40(d,J=7.7Hz,1H),7.09(dd,J=16.2,8.3Hz,2H),7.06(d,J=8.1Hz,1H),6.92(d,J=29 .6Hz,1H),6.73(d,J=8.4Hz,1H),6.10(s,1H),4.55(d,J=22.6Hz,2H),3.10(s,4H),2.83-2.53 (m,4H),2.50-2.38(m,4H),2.35(s,2H),2.10(s,3H),2.06-1.93(m,2H),0.87(d,J=4.9Hz,3H).

[0179] Example 5 Preparation of 2-(6-chloro-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (5)

[0180] Replacing 2,3-diaminopyridine with 5-fluoropyridine-2,3-diamine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 5. LRMS(ESI) m / z 567 (M+). 1 H NMR (600MHz, DMSO) δ10.04(s,1H),8.45(s,1H),8.26(s,1H),7.51(d,J=8.0Hz,2H),7.39(d,J=7.3Hz,2H),7.05(d,J=7.8Hz,1H),6.92(s,1H),6.72 (d,J=8.2Hz,1H),6.08(s,1H),4.51(s,2H),3.12(d,J=43.0Hz,4H),2.49 -2.35(m,4H),2.09(s,3H),2.01(d,J=12.8Hz,4H),0.86(t,J=7.1Hz,4H).

[0181] Example 6 Preparation of 2-(2-((4-fluorobenzyl)thio)-6-(trifluoromethyl)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (6) The 2,3-diaminopyridine was replaced with 5-bromopyridine-2,3-diamine. The other required raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 6. LRMS(ESI) m / z 533 (M+).

[0182] Example 7 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (7)

[0183] Replacing N-methylpiperazine with 3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester, the remaining raw materials, reagents, and preparation methods were the same as in Example 1. Finally, the protecting tert-butoxy carbonyl group was removed by adding 1,4-dioxane hydrochloride in dichloromethane as a solvent to obtain product 7. LRMS(ESI) m / z 531 (M+).

[0184] Example 8 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(6-chloro-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (8)

[0185] Replace 2,3-diaminopyridine with 5-chloropyridine-2,3-diamine; the remaining raw materials, reagents, and preparation methods are the same as in Example 7. LRMS(ESI) m / z 565 (M+).

[0186] Example 9 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (9)

[0187] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine, and using the same raw materials, reagents, and preparation method as in Example 7, yielded product (10). LRMS(ESI) m / z 609 (M+).

[0188] Example 10 Preparation of 2-(2-((4-fluorobenzyl)thio)-7-methyl-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (10)

[0189] Replacing 2,3-diaminopyridine with 3-methylphenyl-1,2-diamine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 10. LRMS(ESI) m / z 547 (M+).

[0190] Example 11 Preparation of 2-(6-cyano-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (11)

[0191] Replacing 2,3-diaminopyridine with 3,4-diaminobenzyl nitrile, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 11. LRMS(ESI) m / z 558 (M+).

[0192] Example 12 Preparation of 2-(2-((4-fluorobenzyl)thio)-6-methoxy-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (12)

[0193] Replacing 2,3-diaminopyridine with 4-methoxyphenyl-1,2-diamine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 12. LRMS(ESI) m / z 563 (M+).

[0194] Example 13 Preparation of 2-(2-(benzylthio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(4-ethylpiperazin-1-yl)-2-methylphenyl)butyramide (13)

[0195] Replacing N-methylpiperazine with 1-ethylpiperazine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 13. LRMS(ESI) m / z 529 (M+).

[0196] Example 14 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(4-isopropylpiperazin-1-yl)-2-methylphenyl)butyramide (14)

[0197] Replacing N-methylpiperazine with 1-isopropylpiperazine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 14. LRMS(ESI) m / z 561 (M+).

[0198] Example 15 Preparation of N-(2-methyl-5-(4-(2,2,2-trifluoroethyl)piperazin-1-yl)phenyl)-2-(2-((3-methylbenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (15)

[0199] Replacing N-methylpiperazine with 1-(2,2,2-trifluoroethyl)piperazine and p-fluorobenzyl bromide with 1-(bromomethyl)-3-methylbenzene, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 15. LRMS(ESI) m / z 597 (M+).

[0200] Example 16 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(4-hydroxypiperidin-1-yl)-2-methylphenyl)butyramide (16)

[0201] Replacing N-methylpiperazine with piperidin-4-ol, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 15. LRMS(ESI) m / z 534 (M+).

[0202] Example 17 Preparation of N-(5-(4-cyanopiperazin-1-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (17)

[0203] Replacing N-methylpiperazine with piperazine-1-onitrile, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 17. LRMS(ESI) m / z 544 (M+).

[0204] Example 18 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-morpholinophenyl)butyramide (18)

[0205] Replacing N-methylpiperazine with morpholine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 18. LRMS(ESI) m / z 520 (M+). 1 H NMR (600MHz, DMSO) δ10.01(s,1H),8.24(d,J=5.5Hz,1H),8.04(d,J=7.2Hz,1H),7.54(d,J=4.8Hz,2H),7.18(d,J=6.8Hz,1H),7.14-6.96(m,3H),6 .94(s,1H),6.72(d,J=8.0Hz,1H),6.11(s,1H),4.55(s,2H),3.67(s,4H) ,2.97(s,4H),2.39(d,J=4.4Hz,2H),2.09(s,3H),0.87(d,J=6.5Hz,3H).

[0206] Example 19 Preparation of 2-(6-chloro-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-morpholinophenyl)butyramide (19)

[0207] Replace 2,3-diaminopyridine with 5-chloropyridine-2,3-diamine, and N-methylpiperazine with morpholine. The other required raw materials, reagents and preparation methods are the same as in Example 1, and product 19 is obtained. 1 H NMR (400MHz, CDCl3) δ8.90 (s, 1H), 7.89 (d, J = 3.7Hz, 2H), 7.52 (d, J = 1.9Hz, 1H), 7.41 (dd,J=7.7,5.9Hz,2H),6.97(dd,J=16.4,8.4Hz,3H),6.60(dd,J=8.4,2.4Hz,1H),5. 80(t,J=7.6Hz,1H),4.56(s,2H),3.85-3.72(m,4H),3.10-3.02(m,4H),2.57(dq,J=1 5.0,7.3Hz,1H),2.28-2.11(m,1H),2.04(s,3H),1.07(t,J=7.3Hz,3H).LRMS(ESI)m / z 554(M+).

[0208] Example 20 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-morpholinophenyl)butyramide (20)

[0209] Replace 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine, replace N-methylpiperazine with morpholine, and use the same raw materials, reagents and preparation methods as in Example 1 to obtain product 20. 1 H NMR (600MHz, DMSO) δ9.97 (s, 1H), 8.46 (s, 1H), 8.26 (d, J = 1.2Hz, 1H), 7.53 (dd, J = 8.5, 5.6Hz, 2H), 7.12-7.02 (m, 3H), 6.94 (s, 1H), 6.71 (d, J = 8. 4Hz,1H),6.09(dd,J=9.3,5.9Hz,1H),4.57(s,2H),3.70-3.64(m,4H),3 .01-2.95(m,4H),2.48-2.37(m,2H),2.10(s,3H),0.89(t,J=7.2Hz,3H).

[0210] Example 21 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(7H-pyrrolo[2,3-b]pyridin-7-yl)butyramide (21)

[0211] Intermediate 1.2 was replaced with 1H-pyrrolo[2,3-b]pyridine, and the remaining raw materials, reagents, and preparation methods were the same as in Example 7, yielding product 21. LRMS(ESI) m / z 390 (M+).

[0212] Example 22 Preparation of N-(2-methyl-5-morpholinophenyl)-2-(7H-pyrrolo[2,3-b]pyridin-7-yl)butyramide (22)

[0213] Intermediate 1.2 was replaced with 1H-pyrrolo[2,3-b]pyridine, and N-methylpiperazine was replaced with morpholine. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 22. LRMS(ESI) m / z 379 (M+). 1H NMR (600MHz, DMSO) δ9.76 (s, 1H), 8.26 (s, 1H), 7.99 (d, J = 7.6Hz, 1H), 7.76 (s ,1H),7.14-7.10(m,1H),7.04(d,J=8.0Hz,1H),6.96(s,1H),6.69(d,J=8.3H z,1H),6.53(s,1H),5.75(t,J=7.1Hz,1H),3.68(s,4H),2.97(s,4H),2.26(d t,J=13.3,6.6Hz,1H),2.19-2.12(m,1H),2.06(s,3H),0.84(t,J=6.4Hz,3H).

[0214] Example 23 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(4H-imidazo[4,5-b]pyridin-4-yl)butyramide (23)

[0215] Intermediate 1.2 was replaced with 1H-imidazo[4,5-b]pyridine, and the remaining raw materials, reagents, and preparation methods were the same as in Example 7, yielding product 23. LRMS(ESI) m / z 391 (M+).

[0216] Example 24 Preparation of 2-(4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-morpholinophenyl)butyramide (24)

[0217] Intermediate 1.2 was replaced with 1H-imidazo[4,5-b]pyridine, and N-methylpiperazine was replaced with morpholine. The remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 24. LRMS(ESI) m / z 380 (M+). 1 H NMR (600MHz, DMSO) δ12.65(s,1H),10.07(s,1H),8.32(s,1H),7.56(s,1H),7.07(d,J=8.1Hz,1H),6.97(s,1H),6.74(d,J=8 .1Hz,1H),5.98(t,J=7.0Hz,1H),3.16(s,3H),2.76(s,3H),2.39(dd,J=23.5,16.9Hz,5H),2.11(s,3H),0.91-0.84(m,3H).

[0218] Example 25 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(6-methyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)phenyl)butyramide (25)

[0219] Replacing N-methylpiperazine with 6-methyl-3,6-diazabicyclo[3.1.1]heptane, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 25. LRMS(ESI) m / z 545 (M+).

[0220] Example 26 Preparation of 3-(4-(3-(2-((cyclopentaeno-1,3-dien-1-methyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butamido)-4-methylphenyl)piperazin-1-yl)propionic acid (26)

[0221] Replacing p-fluorobenzyl bromide with 2-(bromomethyl)thiophene, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 26. LRMS(ESI) m / z 561 (M+).

[0222] Example 27 Preparation of N-(5-(4-(2-(dimethylamino)ethyl)piperazin-1-yl)-2-methylphenyl)-2-(2-(thiophen-2-ylmethyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (27)

[0223] Replacing p-fluorobenzyl bromide with 2-(bromomethyl)thiophene and N-methylpiperazine with N,N-dimethyl-2-(piperazin-1-yl)ethane-1-amine, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 27. LRMS(ESI) m / z 578 (M+).

[0224] Example 28 Preparation of 2-(4-(3-(2-(2-(benzylthio)-4H-imidazo[4,5-b]pyridin-4-yl)-2-cyclopropylacetamido)-4-methylphenyl)piperazin-1-yl)ethane-1-sulfonic acid (28)

[0225] Replacing p-fluorobenzyl bromide with (bromomethyl)benzene and N-methylpiperazine with 2-(piperazin-1-yl)ethane-1-sulfonic acid, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 28. LRMS(ESI) m / z 621 (M+).

[0226] Example 29 Preparation of 2-(2-((4-(furan-2-yl)benzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (29)

[0227] Replacing p-fluorobenzyl bromide with 2-(4-(bromomethyl)phenyl)furan, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 29. LRMS(ESI) m / z 581 (M+).

[0228] Example 30 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methyl-3-phenylpiperazin-1-yl)phenyl)butyramide (30)

[0229] Replacing N-methylpiperazine with 1-methyl-2-phenylpiperazine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 30. LRMS(ESI) m / z 609 (M+).

[0230] Example 31: Preparation of N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)-2-(2-((4-(trifluoromethyl)benzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (31)

[0231] Replacing p-fluorobenzyl bromide with 1-(bromomethyl)-4-(trifluoromethyl)benzene, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 31. LRMS(ESI) m / z 583 (M+).

[0232] Example 32 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-((1R,5S)-3-methyl-3,8-diazabicyclo[3.2.1]octane-8-yl)phenyl)butyramide (32)

[0233] Replacing N-methylpiperazine with (1R,5S)-3-methyl-3,8-diazabicyclo[3.2.1]octane, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 32. LRMS(ESI) m / z 559 (M+).

[0234] Example 33 Preparation of 2-(2-((4-cyanobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(piperazin-1-yl)phenyl)butyramide (33)

[0235] Replacing p-fluorobenzyl bromide with 4-(bromomethyl)benzyl nitrile and N-methylpiperazine with piperazine-1-carboxylic acid tert-butyl ester, the remaining raw materials, reagents, and preparation methods were the same as in Example 7, yielding product 33. LRMS(ESI) m / z 545 (M+).

[0236] Example 34 Preparation of 2-(2-((4-cyanobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(3-(cyanomethyl)-4-methylpiperazin-1-yl)-2-methylphenyl)butyramide (34)

[0237] Replacing p-fluorobenzyl bromide with 4-(bromomethyl)benzyl nitrile and N-methylpiperazine with 2-(1-methylpiperazin-2-yl)acetonitrile, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 34. LRMS(ESI) m / z 579 (M+).

[0238] Example 35 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(6-methyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)phenyl)butyramide (35)

[0239] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and N-methylpiperazine with 6-methyl-3,6-diazabicyclo[3.1.1]heptane, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 35. LRMS(ESI) m / z 623 (M+).

[0240] Example 36 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-(benzylthio)-6-bromo-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (36)

[0241] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and p-fluorobenzyl bromide with (bromomethyl)benzene, the remaining raw materials, reagents, and preparation methods were the same as in Example 7, yielding product 36. LRMS(ESI) m / z 591 (M+).

[0242] Example 37 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(4-(methoxymethyl)piperazin-1-yl)-2-methylphenyl)butyramide (37)

[0243] Replacing N-methylpiperazine with 1-(methoxymethyl)piperazine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 36. LRMS(ESI) m / z 563 (M+).

[0244] Example 38 Preparation of N-(5-(4-(cyanomethyl)piperazin-1-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (38)

[0245] Replacing N-methylpiperazine with 2-(piperazin-1-yl)acetonitrile, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 38. LRMS(ESI) m / z 558 (M+).

[0246] Example 39 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methyl-1,4-aza-1-yl)phenyl)butyramide (39)

[0247] Replacing N-methylpiperazine with 1-methyl-1,4-diazacyclohexane, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 39. LRMS(ESI) m / z 547 (M+).

[0248] Example 40 Preparation of N-(5-(1,4-diaza-1-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (40)

[0249] Replacing N-methylpiperazine with tert-butyl 1,4-diaza-1-carboxylic acid, and using the same raw materials, reagents, and preparation method as in Example 7, yielded product 40. LRMS(ESI) m / z 533 (M+).

[0250] Example 41 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(5-methyl-2,5-diazabicyclo[2.2.1]heptane-2-yl)phenyl)butyramide (41)

[0251] Replacing N-methylpiperazine with 2-methyl-2,5-diazabicyclo[2.2.1]heptane, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 41. LRMS(ESI) m / z 545 (M+).

[0252] Example 42 Preparation of 2-(2-(benzylthio)-4H-imidazo[4,5-b]pyridin-4-yl)-3-methyl-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (42)

[0253] Ethyl 2-bromobutyrate was replaced with 2-bromo-3-methylbutyric acid, and the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 42. LRMS(ESI) m / z 529 (M+).

[0254] Example 43 Preparation of 2-(2-((4-cyclopropylbenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (43)

[0255] Replacing p-fluorobenzyl bromide with 1-(bromomethyl)-4-cyclopropylbenzene, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 43. LRMS(ESI) m / z 555 (M+).

[0256] Example 44 Preparation of N-(5-(4-(dimethylamino)piperidin-1-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (44)

[0257] Replacing N-methylpiperazine with N,N-dimethylpiperidin-4-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 44. LRMS(ESI) m / z 561 (M+).

[0258] Example 45 Preparation of 3,3,3-trifluoro-2-(2-((4-methoxybenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(piperazin-1-yl)phenyl)propionamide (45)

[0259] Replacing p-fluorobenzyl bromide with 1-(bromomethyl)-4-methoxybenzene, ethyl 2-bromobutyrate with 2-bromo-3,3,3-trifluoropropionic acid, and N-methylpiperazine with piperazine-1-carboxylic acid tert-butyl ester, the remaining raw materials, reagents, and preparation methods were the same as in Example 7, yielding product 45. LRMS(ESI) m / z 571 (M+).

[0260] Example 46 Preparation of N-(5-(4-aminopiperidin-1-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (46)

[0261] Replacing N-methylpiperazine with piperidine-4-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 46. LRMS(ESI) m / z 533 (M+).

[0262] Example 47 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(2-methylpiperazin-1-yl)phenyl)butyramide (47)

[0263] Replacing N-methylpiperazine with tert-butyl 3-methylpiperazine-1-carboxylate, and using the same raw materials, reagents, and preparation method as in Example 7, yielded product 47. LRMS(ESI) m / z 533 (M+).

[0264] Example 48 Preparation of 2-(2-((4-cyanobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-aminosulfonylpiperazin-1-yl)phenyl)pentanamide (48)

[0265] Replacing p-fluorobenzyl bromide with 4-(bromomethyl)benzyl nitrile, ethyl 2-bromobutyrate with ethyl 2-bromopentanoate, and N-methylpiperazine with piperazine-1-sulfonamide, while using the same raw materials, reagents, and preparation method as in Example 1, yielded product 48. LRMS(ESI) m / z 619 (M+).

[0266] Example 49 Preparation of 2-(2-(benzylthio)-6-bromo-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-morpholinophenyl)butyramide (49)

[0267] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine, p-fluorobenzyl bromide with (bromomethyl)benzene, and N-methylpiperazine with morpholine, while using the same raw materials, reagents, and preparation method as in Example 1, yielded product 49. LRMS(ESI) m / z 580 (M+).

[0268] Example 50 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(6-bromo-2-((4-methoxybenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (50)

[0269] The 2,3-diaminopyridine was replaced with 5-bromopyridine-2,3-diamine, p-fluorobenzyl bromide was replaced with 1-(bromomethyl)-4-methoxybenzene, and N-methylpiperazine was replaced with 3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester. The remaining raw materials, reagents, and preparation methods were the same as in Example 7, yielding product 50. LRMS(ESI) m / z 621 (M+).

[0270] Example 51 Preparation of 2-(6-bromo-2-mercapto-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (51)

[0271] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and p-fluorobenzyl bromide with 1-(bromomethyl)-4-methoxybenzene, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding an intermediate. The intermediate was dissolved in trifluoroacetic acid, followed by the addition of m-cresol, and the mixture was refluxed at 70°C for 18 hours. After the reaction, the pH was adjusted to neutral with saturated sodium bicarbonate solution, diluted with water, and extracted with ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Column chromatography was performed using dichloromethane / methanol = 25 / 1-20 / 1 (v / v) as the eluent to obtain product 51, with a yield of 42%. LRMS(ESI) m / z 502 (M+).

[0272] Example 52 Preparation of N-(5-(4-(dimethylamino)piperidin-1-yl)-2-methylphenyl)-2-(2-(((4-ethylbenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)propionamide (52)

[0273] Replacing p-fluorobenzyl bromide with 1-(bromomethyl)-4-ethylbenzene and N-methylpiperazine with N,N-dimethylpiperidin-4-amine, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 52. LRMS(ESI) m / z 557 (M+).

[0274] Example 53 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(imidazolidine-1-yl)-2-methylphenyl)butyramide (53)

[0275] Replacing N-methylpiperazine with imidazolidine-1-carboxylic acid tert-butyl ester, and using the same raw materials, reagents, and preparation method as in Example 7, yielded product 53. LRMS(ESI) m / z 505 (M+).

[0276] Example 54 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(3-methylimidazolidine-1-yl)phenyl)butyramide (54)

[0277] Replacing N-methylpiperazine with 1-methylimidazolidine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 54. LRMS(ESI) m / z 519 (M+).

[0278] Example 55 Preparation of N-(5-(4-(dimethylamino)piperazin-1-yl)-2-methylphenyl)-2-(2-(((2-fluorobenzyl)thio)-5,6-dimethyl-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (55)

[0279] Replacing 2,3-diaminopyridine with 5,6-dimethylpyridine-2,3-diamine, p-fluorobenzyl bromide with 1-(bromomethyl)-2-fluorobenzene, and N-methylpiperazine with N,N-dimethylpiperazine-1-amine, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 55. LRMS(ESI) m / z 590 (M+).

[0280] Example 56 Preparation of 2-(5,6-difluoro-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(piperidin-4-yl)phenyl)butyramide (56)

[0281] Replacing 2,3-diaminopyridine with 5,6-difluoropyridine-2,3-diamine and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with tert-butyl 4-(3-amino-4-methylphenyl)piperidine-1-carboxylate, the remaining raw materials, reagents, and preparation methods were the same as in Example 7, yielding product 56. LRMS(ESI) m / z 554 (M+).

[0282] Example 57 Preparation of 2-(2-((3-(furan-2-yl)benzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(1-methylpiperidin-4-yl)phenyl)butyramide (57)

[0283] Replacing p-fluorobenzyl bromide with 2-(3-(bromomethyl)phenyl)furan, and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 2-methyl-5-(1-methylpiperidin-4-yl)aniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 57. LRMS(ESI) m / z 580 (M+).

[0284] Example 58 Preparation of 2-(2-((4-chlorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(1-methyl-1,2,3,6-tetrahydropyridin-4-ylphenyl)butyramide (58)

[0285] Replacing 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 2-methyl-5-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)aniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 58. LRMS(ESI) m / z 546 (M+).

[0286] Example 59 Preparation of 2-(2-((4-bromobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(7-methyl-4,7-diazaspiro[2,5]oct-4-yl)phenyl)butyramide (59)

[0287] Replacing N-methylpiperazine with 7-methyl-4,7-diazaspiro[2.5]octane, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 59. LRMS(ESI) m / z 619 (M+).

[0288] Example 60 Preparation of N-(5-(6-(cyanomethyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (60)

[0289] Replacing N-methylpiperazine with 2-(3,6-diazabicyclo[3.1.1]heptane-6-yl)acetonitrile, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 60. LRMS(ESI) m / z 570 (M+).

[0290] Example 61: Preparation of N-(5-(6-cyano-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (61)

[0291] Replacing N-methylpiperazine with 3,6-diazabicyclo[3.1.1]heptane-6-nitrile, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 61. LRMS(ESI) m / z 556 (M+).

[0292] Example 62 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-(3-fluoropropyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (62)

[0293] Compound 7 (100 mg, 0.164 mmol) was dissolved in acetonitrile, followed by the addition of potassium carbonate (66.40 mg, 0.656 mmol) and 1-fluoro-3-iodopropane (61.68 mg, 0.328 mmol). The reaction was refluxed at 50 °C for 15 hours. After the reaction was completed, the mixture was extracted with ethyl acetate / water. The organic phases were combined, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was separated by column chromatography using dichloromethane / methanol = 20 / 1 (v / v) as the eluent to obtain product 62, with a yield of 32%. LRMS (ESI) m / z 559 (M+). 1 H NMR (600MHz, DMSO) δ10.02(s,1H),8.46(s,1H),8.27(s,1H),7.54(s,2H),7.09(t,J=7.8Hz, 2H),7.05(d,J=7.7Hz,1H),6.74(s,1H),6.50(d,J=7.3Hz,1H),6.11(s,1H),4.56(s,2H),4. 48(s,1H),4.40(s,1H),3.67(s,2H),3.20(s,2H),2.49-2.37(m,4H),2.34(s,2H),2.09(s,3 H), 2.00 (dd, J = 16.5, 7.7Hz, 1H), 1.66 (s, 2H), 1.48 (d, J = 35.0Hz, 1H), 0.88 (t, J = 6.2Hz, 3H).

[0294] Example 63 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(6-(2,2,2-trifluoroethyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)phenyl)butyramide (63)

[0295] Compound 7 (100 mg, 0.164 mmol) was dissolved in dichloromethane, followed by the addition of trifluoromethyltrifluoromethanesulfonate (42.93 mg, 0.197 mmol) and triethylamine (66.40 mg, 0.656 mmol). The reaction was carried out at room temperature for 2 hours. After the reaction was completed, the solution was concentrated under reduced pressure to obtain a brown crude product. The product 63 was obtained by column chromatography using dichloromethane / methanol = 20 / 1 (v / v) as the eluent, with a yield of 31%. LRMS (ESI) m / z 691 (M+).

[0296] Example 64 Preparation of 3-(4-methyl-3-(2-(((4-nitrobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butamido)phenyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid (64)

[0297] Replacing p-fluorobenzyl bromide with 1-(bromomethyl)-4-nitrobenzene and N-methylpiperazine with 3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 64. LRMS(ESI) m / z 602(M+).

[0298] Example 65 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(1,7-diazaspiro[4,4]non-7-yl)phenyl)butyramide (65)

[0299] Replacing N-methylpiperazine with 1,7-diazaspiro[4.4]nonane, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 65. LRMS(ESI) m / z 559 (M+).

[0300] Example 66 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(6-(oxecyclobutane-3-yl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)phenyl)butyramide (66)

[0301] Replacing N-methylpiperazine with 6-(oxecyclobutan-3-yl)-3,6-diazabicyclo[3.1.1]heptane, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 66. LRMS(ESI) m / z 587 (M+).

[0302] Example 67 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-hydroxy-3-azabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (67)

[0303] Replacing N-methylpiperazine with 3-azabicyclo[3.1.1]heptane-6-ol, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 67. LRMS(ESI) m / z 546 (M+).

[0304] Example 68 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-(hydroxymethyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (68)

[0305] Replacing N-methylpiperazine with (3,6-diazabicyclo[3.1.1]heptane-6-yl)methanol, with the remaining raw materials, reagents, and preparation methods the same as in Example 1, yielded product 68. LRMS(ESI) m / z 561 (M+).

[0306] Example 69 Preparation of N-(5-(6-(dimethylamino)-3-azabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (69)

[0307] Replacing N-methylpiperazine with N,N-dimethyl-3-azabicyclo[3.1.1]heptane-6-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 69. LRMS(ESI) m / z 573 (M+).

[0308] Example 70 Preparation of N-(5-(6-(dimethylamino)-3-azabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (70)

[0309] Replacing N-methylpiperazine with 3-(3,6-diazabicyclo[3.1.1]heptane-6-yl)-N,N-dimethylpropyl-1-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 70. LRMS(ESI) m / z 616 (M+).

[0310] Example 71: Preparation of N-(5-(6-(4-(dimethylamino)butyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (71)

[0311] Replacing N-methylpiperazine with 4-(3,6-diazabicyclo[3.1.1]heptane-6-yl)-N,N-dimethylbut-1-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 71. LRMS(ESI) m / z 630 (M+).

[0312] Example 72 Preparation of 2-(2-((4-(dimethylamino)benzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-methylpiperazin-1-yl)phenyl)butyramide (72)

[0313] Replacing p-fluorobenzyl bromide with 4-(bromomethyl)-N,N-dimethylaniline, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 72. LRMS(ESI) m / z 558 (M+).

[0314] Example 73 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-methoxy-3-azabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (73)

[0315] Replacing N-methylpiperazine with 6-methoxy-3-azabicyclo[3.1.1]heptane, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 73. LRMS(ESI) m / z 560 (M+).

[0316] Example 74 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(6-(methylamino)-3-azabicyclo[3.1.1]heptane-3-yl)phenyl)butyramide (74)

[0317] Replacing N-methylpiperazine with N-methyl-3-azabicyclo[3.1.1]heptane-6-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 74. LRMS(ESI) m / z 559 (M+).

[0318] Example 75 Preparation of N-(5-(6-(dimethylamino)bicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (75)

[0319] Replacing N-methylpiperazine with 3-(3-amino-4-methylphenyl)-N,N-dimethylbicyclo[3.1.1]heptane-6-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 75. LRMS(ESI) m / z 572 (M+).

[0320] Example 76 Preparation of N-(5-(3,6-dihydro-2H-pyran-4-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (76)

[0321] Replacing N-methylpiperazine with 5-(3,6-dihydro-2H-pyran-4-yl)-2-methylaniline, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 76. LRMS(ESI) m / z 517 (M+).

[0322] Example 77 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(tetrahydro-2H-pyran-4-yl)phenyl)butyramide (77)

[0323] Replacing N-methylpiperazine with 2-methyl-5-(tetrahydro-2H-pyran-4-yl)aniline, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 77. LRMS(ESI) m / z 519 (M+).

[0324] Example 78 Preparation of N-(5-(6-cyclopropyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (78)

[0325] Replacing N-methylpiperazine with 6-cyclopropyl-3,6-diazabicyclo[3.1.1]heptane, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 78. LRMS(ESI) m / z 571 (M+).

[0326] Example 79 Preparation of N-(5-(6-amino-3-azabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (79)

[0327] Replacing N-methylpiperazine with 3-azabicyclo[3.1.1]heptane-6-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 79. LRMS(ESI) m / z 545 (M+).

[0328] Example 80 Preparation of N-(5-(6-amino-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (80)

[0329] Replacing N-methylpiperazine with 3,6-diazabicyclo[3.1.1]heptane-6-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 80. LRMS(ESI) m / z 546 (M+).

[0330] Example 81: Preparation of N-(5-(azacyclopentan-4-yl)-2-methylphenyl)-2-(2-(((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (81)

[0331] Replacing N-methylpiperazine with tert-butyl 4-(3-amino-4-methylphenyl)aza-1-carboxylic acid, and using the same raw materials, reagents, and preparation method as in Example 7, yielded product 81. LRMS(ESI) m / z 532 (M+).

[0332] Example 82: Preparation of N-(5-(3-aminocyclopentyl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (82)

[0333] Replacing N-methylpiperazine with 5-(3-aminocyclopentyl)-2-methylaniline, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 82. LRMS(ESI) m / z 518 (M+).

[0334] Example 83 Preparation of N-(5-(4-aminopiperazin-1-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (83)

[0335] Replacing N-methylpiperazine with 4-(3-amino-4-methylphenyl)piperazine-1-amine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 83. LRMS(ESI) m / z 534 (M+).

[0336] Example 84 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-(3-hydroxy-3-methylbutyryl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (84)

[0337] Compound 8 (150 mg, 0.246 mmol) was dissolved in dichloromethane, followed by the addition of triethylamine (127.22 mg, 0.984 mmol) and HATU (140.35 mg, 0.369 mmol). The mixture was stirred at room temperature for 10 minutes, and then 4-hydroxy-4-methylpentanoic acid (26.02 mg, 0.197 mmol) was added dropwise. The reaction was continued at room temperature for 2 hours. After the reaction was completed, the mixture was filtered and concentrated under reduced pressure to obtain a brown crude product. Column chromatography was performed using dichloromethane / methanol = 20 / 1 (v / v) as the eluent to obtain product 84, with a yield of 37%. LRMS(ESI) m / z 709 (M+). 1H NMR (600MHz, DMSO) δ10.02(d,J=7.7Hz,1H),8.47(s,1H),8.27(s,1H),7.54(s,2H),7.09(t,J=8.2Hz,2H),7.03(d,J=8. 0Hz,1H),6.70(d,J=17.1Hz,1H),6.47(d,J=8.0Hz,1H),6.10(d,J=4.8Hz,1H),4.72(s,1H),4.63(s,1H),4.56(s,2H),4. 39(s,1H),3.59(d,J=10.4Hz,1H),3.50(d,J=10.5Hz,1H),3.29(d,J=10.3Hz,1H),2.58(s,1H),2.44(s,2H),2.28(d,J=1 3.8Hz,1H),2.06(d,J=13.4Hz,4H),2.04-1.91(m,1H),1.52(d,J=8.1Hz,1H),1.09(d,J=17.1Hz,6H),0.89-0.85(m,3H).

[0338] Example 85 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-(ethylsulfonyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (85)

[0339] Compound 8 (100 mg, 0.164 mmol) was dissolved in dichloromethane, followed by the addition of triethylamine (66.40 mg, 0.656 mmol). Ethylenesulfonyl chloride (21.09 mg, 0.164 mmol) was then added dropwise under ice bath conditions. The reaction was carried out at room temperature for 12 hours. After the reaction was complete, the solution was concentrated under reduced pressure to obtain a brown crude product. Column chromatography was performed using dichloromethane / methanol = 20 / 1 (v / v) as the eluent to obtain product 85, with a yield of 34%. LRMS (ESI) m / z 701 (M+). 1H NMR(600MHz,DMSO)δ10.01(s,1H),8.48(s,1H),8.28(s,1H),7.54(s,2H),7 .15-7.01(m,3H),6.73(s,1H),6.49(s,1H),6.10(s,1H),4.57(s,2H),4.40 (s,2H),3.60(s,2H),3.44(s,2H),3.07(s,2H),2.81(s,1H),2.48-2.33(m, 2H), 2.09 (s, 3H), 1.64 (s, 1H), 1.17 (d, J = 3.3Hz, 3H), 0.88 (d, J = 3.7Hz, 3H).

[0340] Example 86 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-(2-methoxyethyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (86)

[0341] Compound 8 (100 mg, 0.164 mmol) was dissolved in DMF, followed by the addition of 1-bromo-2-methoxyethane (66.40 mg, 0.656 mmol). Ethylenesulfonyl chloride (21.09 mg, 0.164 mmol) was then added dropwise under ice bath conditions. The reaction was carried out at room temperature for 12 hours. After the reaction was complete, the solution was concentrated under reduced pressure to obtain a brown crude product. Product 86 was obtained by column chromatography using dichloromethane / methanol = 20 / 1 (v / v) as the eluent, with a yield of 37%. LRMS (ESI) m / z 701 (M+).

[0342] Example 87 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(6-(oxecyclobutane-3-yl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)phenyl)butyramide (87)

[0343] Replacing N-methylpiperazine with 1-(oxecyclobutane-3-yl)piperazine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 87. LRMS(ESI) m / z 575 (M+).

[0344] Example 88 Preparation of 2-(6-chloro-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-(oxecyclobutane-3-yl)piperazin-1-yl)phenyl)butyramide (88)

[0345] Replacing 2,3-diaminopyridine with 5-chloropyridine-2,3-diamine and N-methylpiperazine with 1-(oxecyclobutane-3-yl)piperazine, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 88. LRMS(ESI) m / z 609 (M+).

[0346] Example 89 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(4-(oxecyclobutane-3-yl)piperazin-1-yl)phenyl)butyramide (89)

[0347] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and N-methylpiperazine with 1-(oxecyclobutane-3-yl)piperazine, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 89. LRMS(ESI) m / z 653 (M+). 1 HNMR(600MHz,DMSO)δ9.99(s,1H),8.46(s,1H),8.27(s,1H),7.53(s,2H), 7.09(t,J=8.0Hz,2H),7.04(d,J=8.1Hz,1H),6.92(s,1H),6.72(d,J=8.3Hz ,1H),6.07(s,1H),4.55(d,J=11.1Hz,4H),4.43(s,2H),3.39(s,1H),3.03( s,4H),2.48-2.37(m,2H),2.33(s,4H),2.08(s,3H),0.86(t,J=5.4Hz,3H).

[0348] Example 90 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-isopropyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (90)

[0349] Compound 7 (100 mg, 0.164 mmol) was dissolved in MeOH / AcOH = 4 / 1 (v / v), followed by the addition of acetone (21.89 mg, 0.377 mmol) and sodium cyanoborohydride (59.21 mg, 0.942 mmol). The mixture was refluxed at 50 °C for 12 hours. After the reaction, the pH was adjusted to weakly alkaline with saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate / water, and the organic phase was concentrated to obtain a brown crude product. Column chromatography was performed using dichloromethane / methanol = 20 / 1 (v / v) as eluent to obtain product 90, with a yield of 37%. LRMS (ESI) m / z 573 (M+). 1H NMR (600MHz, CDCl3) δ8.89(s,1H),7.97(d,J=7.5Hz,1H),7.76(d,J=6.4Hz,1H),7.47(d,J=7.8Hz,1 H),7.44(t,J=5.8Hz,2H),7.07(t,J=7.0Hz,1H),6.99(dd,J=16.9,8.1Hz,3H),6.40(d,J=8.3Hz,1H ),5.83(s,1H),4.60(s,2H),3.85(s,2H),3.49(d,J=10.9Hz,2H),3.28(s,2H),2.69-2.57(m,3H),2 .26(dt,J=13.4,6.8Hz,1H),2.06(s,3H),1.57(d,J=8.4Hz,1H),1.09(t,J=7.0Hz,3H),0.96(s,6H).

[0350] Example 91 Preparation of 2-(6-chloro-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-isopropyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (91)

[0351] Replacing compound 7 with 8, and using the same raw materials, reagents, and preparation method as in Example 90, yielded product 91. LRMS(ESI) m / z 607 (M+). 1 H NMR (600MHz, DMSO) δ10.02(s,1H),8.46(s,1H),8.19(s,1H),7.54(s,2H),7.13-7.01(m,3H),6.71(d,J=31.4Hz,1H),6.49(d,J=7.1Hz,1H), 6.12(s,1H),4.56(s,2H),3.77(s,2H),3.20(s,3H),2.45(dd,J=17.8, 7.7Hz,4H),2.09(s,3H),2.04-1.79(m,1H),1.49(s,1H),0.88(s,9H).

[0352] Example 92 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-isopropyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (92)

[0353] Replacing compound 7 with 9, and using the same raw materials, reagents, and preparation method as in Example 90, yielded product 92. LRMS(ESI) m / z 651 (M+).

[0354] Example 93 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(6-isopropyl-3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)butyramide (93)

[0355] Replacing N-methylpiperazine with 1-cyclopropylpiperazine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 93. LRMS(ESI) m / z 559 (M+). 1 H NMR (600MHz, DMSO) δ9.98 (s, 1H), 8.21 (d, J = 6.4Hz, 1H), 8.03 (d, J = 7.4Hz, 1H), 7.56-7.49 (m, 2H ),7.19(t,J=6.9Hz,1H),7.07(t,J=8.6Hz,2H),7.02(d,J=8.3Hz,1H),6.90(s,1H),6.68(d,J=7. 9Hz,1H),6.07(dd,J=9.1,6.2Hz,1H),4.54(s,2H),2.95(s,4H),2.59(s,4H),2.44-2.29(m,2H) ,2.06(s,3H),1.57(d,J=35.0Hz,1H),0.88(t,J=7.0Hz,3H),0.40(d,J=5.1Hz,2H),0.30(s,2H).

[0356] Example 94 Preparation of 2-(6-chloro-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(4-cyclopropylpiperazin-1-yl)-2-methylphenyl)butyramide (94)

[0357] Replacing 2,3-diaminopyridine with 5-chloropyridine-2,3-diamine and N-methylpiperazine with 1-cyclopropylpiperazine, while using the same raw materials, reagents, and preparation method as in Example 1, yielded product 94. LRMS(ESI) m / z 593 (M+). 1H NMR(600MHz,DMSO)δ9.98(s,1H),8.44(s,1H),8.18(s,1H),7.53(dd,J=8.0,5.9Hz,2 H),7.09(t,J=8.7Hz,2H),7.03(d,J=8.4Hz,1H),6.91(s,1H),6.69(d,J=8.2Hz,1H),6 .09(dd,J=9.6,6.0Hz,1H),4.56(s,2H),2.97(s,4H),2.60(s,4H),2.48-2.33(m,2H) ,2.08(s,3H),1.61(s,1H),0.88(t,J=7.2Hz,3H),0.41(d,J=5.0Hz,2H),0.31(s,2H).

[0358] Example 95 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(4-cyclopropylpiperazin-1-yl)-2-methylphenyl)butyramide (95)

[0359] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and N-methylpiperazine with 1-cyclopropylpiperazine, while using the same raw materials, reagents, and preparation method as in Example 1, yielded a product with 95% LRMS (ESI) m / z 637 (M+). 1 HNMR(400MHz, CDCl3)δ8.88(s,1H),8.17-7.80(m,3H),7.54-7.37(m,4H),7.33-7.19(m ,5H),7.16-6.87(m,2H),6.60(dt,J=15.2,7.6Hz,1H),5.80(t,J=7.7Hz,1H),4.60(s,3H ),3.26-3.00(m,8H),2.86-2.45(m,6H),2.43-2.11(m,3H),2.05(s,4H),1.64(ddd,J=10 .2,6.5,4.0Hz,1H),1.02(t,J=40.6,11.6Hz,5H),0.95-0.80(m,1H),0.54-0.35(m,4H).

[0360] Example 96 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-thiomorpholinyl)butyramide (96)

[0361] Replacing N-methylpiperazine with thiomorpholine, and using the same raw materials, reagents, and preparation method as in Example 1, yielded product 96. LRMS(ESI) m / z 536 (M+).

[0362] Example 97 Preparation of 2-(2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-thiomorpholinyl)butyramide (97)

[0363] Replacing 2,3-diaminopyridine with 5-methylpyridine-2,3-diamine and N-methylpiperazine with 1-(oxecyclobutane-3-yl)piperazine, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 97. LRMS(ESI) m / z 589 (M+). 1 H NMR (600MHz, DMSO) δ9.96(s,1H),8.09(s,1H),7.91(s,1H),7.53(s,2H),7.08(t,J=8.3Hz,2H),7.04(d,J=8.1Hz,1H),6.94(s,1H),6.70(d,J=8.0Hz ,1H),6.05(s,1H),4.54(s,4H),4.43(s,2H),3.38(d,J=11.1Hz,1H),3.03 (s,4H),2.44(s,3H),2.39(s,2H),2.33(s,4H),2.08(s,3H),0.87(s,3H).

[0364] Example 98 Preparation of N-(5-(3,6-diazabicyclo[3.1.1]heptane-3-yl)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-6-methyl-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (98)

[0365] Replacing 2,3-diaminopyridine with 5-methylpyridine-2,3-diamine and N-methylpiperazine with 3,6-diazabicyclo[3.1.1]heptane-6-carboxylic acid tert-butyl ester, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 98. LRMS(ESI) m / z 545 (M+).

[0366] Example 99 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(pyridin-4-yl)phenyl)butyramide (99)

[0367] Replacing 2,3-diaminopyridine with 5-bromopyridin-2,3-diamine and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 2-methyl-5-(pyridin-4-yl)aniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding a product 99. LRMS(ESI) m / z 590 (M+). 1 H NMR (600MHz, DMSO) δ10.21(s,1H),8.58(d,J=5.6Hz,1H),8.49(s,1H),8.28(s,1H),7.78(s,1H),7.61(d,J=5.6Hz,1H),7.57(d,J=7.9Hz,1H),7.53 (dd,J=8.1,5.8Hz,1H),7.38(d,J=7.9Hz,1H),7.08(t,J=8.8Hz,1H),6.12(dd,J=9.5,6.2Hz,1H),4.57(s,1H),2.26(s,1H),0.90(t,J=7.2Hz,1H).

[0368] Example 100 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-methyl-5-(pyridin-4-yl)phenyl)butyramide (100)

[0369] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and N-methylpiperazine with 2-methyloctahydropyrrole[3,4-c]pyrrole, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding a product of 100. LRMS(ESI) m / z 637 (M+). 1 H NMR (600MHz, CDCl3) δ8.85 (s, 1H), 7.99 (s, 2H), 7.43 (s, 2H), 7.27 (d, J = 12.1Hz, 1H), 7.01-6.92(m,3H),6.38(d,J=8.2Hz,1H),5.82(t,J=6.9Hz,1H),4.57(s,2H),3.23-3. 16(m,3H),2.94(s,2H),2.87(s,2H),2.83-2.72(m,1H),2.57(dd,J=13.6,6.7Hz,1H) ,2.41-2.25(m,5H),2.22-2.17(m,1H),2.03(d,J=17.0Hz,3H),1.07(t,J=6.4Hz,3H).

[0370] Example 101 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(3-(pyridin-4-yl)phenyl)butyramide (101)

[0371] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 3-(pyridin-4-yl)aniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 101. LRMS(ESI) m / z 576 (M+).

[0372] Example 102 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(2-cyano-5-(pyridin-4-yl)phenyl)butyramide (102)

[0373] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 2-amino-4-(pyridin-4-yl)benzonitrile, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 102. LRMS(ESI) m / z 601 (M+).

[0374] Example 103 Preparation of N-(2-amino-5-(pyridin-4-yl)phenyl)-2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (103)

[0375] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 4-(pyridin-4-yl)phenyl-1,2-diamine, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 103. LRMS(ESI) m / z 591 (M+).

[0376] Example 104 Preparation of N-(5-(2-(dimethylamino)ethoxy)-2-methylphenyl)-2-(2-((4-fluorobenzyl)thio)-6-(1-isopropyl-1H-pyrazol-5-yl)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (105)

[0377] Replacing 2,3-diaminopyridine with 5-bromopyridine-2,3-diamine and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 5-(2-(dimethylamino)ethoxy)-2-methylaniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 104. LRMS(ESI) m / z 614 (M+).

[0378] Example 105 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(3-(dimethylamino)propoxy)-2-methylphenyl)butyramide (105)

[0379] Replacing 2,3-diaminopyridine with 5-(1-isopropyl-1H-pyrazol-5-yl)pyridine-2,3-diamine, and replacing 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 5-(2-(dimethylamino)ethoxy)-2-methylaniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 105. LRMS(ESI) m / z 630 (M+).

[0380] Example 106 Preparation of N-(5-(2-(dimethylamino)ethoxy)-2-methylphenyl)-2-(6-(1-ethyl-1H-pyrazol-5-yl)-2-((4-fluorobenzyl)oxy)-4H-imidazo[4,5-b]pyridin-4-yl)butyramide (106)

[0381] Replacing 2,3-diaminopyridine with 5-(1-ethyl-1H-pyrazol-5-yl)pyridine-2,3-diamine, replacing p-fluorobenzyl bromide with (4-fluorophenyl)methanol, and replacing 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 5-(2-(dimethylamino)ethoxy)-2-methylaniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 106. LRMS(ESI) m / z 560 (M+).

[0382] Example 107 Preparation of 2-(6-bromo-2-((4-fluorobenzyl)oxy)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(2-(dimethylamino)ethoxy)-2-methylphenyl)butyramide (107)

[0383] Replacing p-fluorobenzyl bromide with (4-fluorophenyl)methanol and 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 5-(2-(dimethylamino)ethoxy)-2-methylaniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 107. LRMS(ESI) m / z 584 (M+).

[0384] Example 108 Preparation of 2-(6-(1-(difluoromethyl)-1H-pyrazol-5-yl)-2-((4-fluorobenzyl)thio)-4H-imidazo[4,5-b]pyridin-4-yl)-N-(5-(3-(dimethylamino)propoxy)-2-methylphenyl)butyramide (108)

[0385] Replacing 2,3-diaminopyridine with 5-(1-(difluoromethyl)-1H-pyrazol-5-yl)pyridine-2,3-diamine, and replacing 2-methyl-5-(4-methylpiperazin-1-yl)aniline with 5-(2-(dimethylamino)propoxy)-2-methylaniline, the remaining raw materials, reagents, and preparation methods were the same as in Example 1, yielding product 108. LRMS(ESI) m / z 652 (M+).

[0386] Example 109 Pharmacological Activity Test Example

[0387] Pharmacological Example 1. Molecular-level inhibitory activity test of the compound against PL protease

[0388] Experimental principle: Based on SARS-CoV-2PL pro Protein is a fundamental characteristic of proteolytic enzymes, and a fluorescence method for detecting PL was established. pro A screening system for protein activity. PL pro Proteins can specifically recognize and cleave diglycine peptides. Activity detection can be performed using fluorescent peptides as substrates, and the activity of proteolytic enzymes can be reflected by detecting the generation of fluorescence signals.

[0389] Experimental results:

[0390] Some compounds are effective against SARS-CoV-2 PL. pro The protease showed some inhibitory effect. The results are shown in Table 1.

[0391] Table 1: Effects of some compounds on PL pro Inhibitory activity of proteases

[0392] compound <![CDATA[PL pro IC 50 (nM)]]> 1 84.2±8.9 5 138.0±7.6 7 47.4±4.4 8 49.9±14 9 34.0±4.0 25 37.0±10.7 35 28.8±2.0 36 41.2±4.0 49 201.2±34.8 50 205.3±28.4 62 179.8±15.0 86 154.1±35.9 88 153.0±10.1 89 106.3±6.1 90 271.4±13.5 91 123.5±16.9 92 215.7±17.1 98 50.4±9.1 100 29.8±7.9

[0393] PL pro Results of protease inhibition experiments showed that, through molecular-level enzyme inhibitory activity screening, several of the aforementioned compounds were found to be effective against SARS-CoV-2 PL. pro The protease has a good inhibitory effect.

[0394] Pharmacological Example 2. Test of the inhibitory activity of the compound against SARS-CoV-2 virus at the cellular level

[0395] Before transfection, Vero E6 cells were cultured and passaged in a CO2 cell culture incubator using DMEM medium containing 10% FBS. For experiments, Vero E6 cells (ATCC-1586) were seeded in 48-well plates at a density of 5000–10000 cells / well. After cell attachment, the supernatant was aspirated, and the cells were incubated in medium containing 10 μL of the compound for 2 hours. Then, SARS-CoV-2 virus strain and Omicron BA.5 coronavirus strain were added, with a multiplicity of infection (MOI) of 0.01–0.1. The cells were co-cultured in a CO2 cell culture incubator for 24 hours. The supernatant was then collected, and viral RNA and reverse RNA were extracted. The viral copy number in the supernatant was detected using real-time quantitative PCR. The inhibitory rate of the compound was calculated based on the viral copy number, and the EC50 of the compound was calculated using Prism 9. 50 All of the above virus experiments were conducted in the BSL-3 biosafety laboratory of the Wuhan Institute of Virology, Chinese Academy of Sciences.

[0396] Experimental results:

[0397] Some compounds showed inhibitory effects on the Omicron BA.5 coronavirus strain of SARS-CoV-2. The results are shown in Table 2.

[0398] Table 2: Inhibitory activity of some compounds against Omicron BA.5 coronavirus strain

[0399] compound Inhibition rate (10 μM) 7 94.0％ 8 96.4％ 9 95.4％ 25 97.3％ 35 95.2％ 36 95.3％ 98 95.3％ 100 95.7％

[0400] The results of the virus replication inhibition experiment showed that the test compounds at a concentration of 10 μM could effectively inhibit the replication of the SARS-CoV-2 virus genome in the infection supernatant, with inhibition rates greater than 90%.

[0401] Pharmacological Example 3. Test of the inhibitory activity of the compound against HCoV-OC43 virus at the cellular level

[0402] Vero E6 cells were cultured in DMEM supplemented with 10% FBS at 37°C and 5% CO2. Before infection, 50,000 Vero E6 cells were seeded in 48-well plates and incubated in DMES (10% FBS) for 12 h. Then, 200 wells were replaced with 1 part DMEM (2% FBS) per well and cultured for 2 h at 10 μM (for initial screening). HCoV-OC43 was then added at an MOI of 0.1. Twenty-four hours post-infection, the supernatant was collected, and viral RNA was extracted and reverse transcribed using the PrimeScript RT kit. To determine viral copy number, TB... Absolute quantitative RT-PCR was performed using Premix Ex Taq™ II (TaKaRa). All experiments involving HCoV-OC43 were conducted in the BSL3 laboratory of the Wuhan Institute of Virology, Chinese Academy of Sciences.

[0403] Experimental results:

[0404] Some compounds showed some inhibitory effect on HCoV-OC43. The results are shown in Table 3.

[0405] Table 3: Inhibitory activity of some compounds against HCoV-OC43

[0406]

[0407]

[0408] Experimental results showed that some of the tested compounds could effectively inhibit the replication of the HCoV-OC43 viral genome.

[0409] All documents mentioned in this invention are incorporated herein by reference as if each document were individually incorporated by reference. Furthermore, it should be understood that after reading the foregoing teachings of this invention, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims.

Claims

1. A compound having a structure shown in Formula I, and a racemate, R-isomer, S-isomer, pharmaceutically acceptable salt thereof, or a mixture thereof: wherein, n and p are each independently 0, 1, 2, 3, or 4; X is selected from the group consisting of CR a , CR a R b , C=0, N, NR a , O, S, S=0, S02; each R a and R b are each independently selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 alkoxy; Y is selected from the group consisting of absent, H, NH2, OH, SH, and Y1is selected from the group consisting of NH, O, S; Ring is selected from the group consisting of saturated or partially saturated C 5-7 carbocyclic, C 6-10 aromatic, 5-6 membered heteroaromatic, 5-7 membered heterocyclic, 6-20 membered heteroaromatic fused ring; wherein the hydrogen on the above cyclic groups is optionally substituted with 1, 2, 3, or 4 substituents selected from the group consisting of halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-8 cycloalkyl, C 3-8 halocycloalkyl, cyano, nitro, amino, C 1-6 amine, hydroxyl, hydroxymethyl, carboxyl, thiol, C 1-6 alkylsulfonyl, C 6-10 aryl, 5-12 membered heteroaryl, 3-12 membered heterocyclyl; each R 1 , R 2 , and R 3 is independently selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 1-6alkyl, C 1-6 1-6haloalkyl, C 1-6 1-6alkoxy, C 1-6 1-6alkoxycarbonyl, C 1-6 1-6haloalkoxy, C 2-6 1-6alkenyl, C 2-6 1-6alkynyl, C 3-8 1-6cycloalkyl, cyano, C 2-6 1-6ester, nitro, amino, C 1-6 1-6amine, hydroxyl, hydroxymethyl, carboxyl, 5-12 membered heteroaryl, 3-12 membered heterocyclyl, -(CH2) p 1-6CONR 7 1-6R 8 , -NR 8 1-6CO(C 1-6 1-6alkyl); m, q, r, s, and t are each independently 0, 1, 2, 3, or 4; Unless otherwise specified, each of the heteroaryl, heteroaromatic ring, hetero-fused ring, heterocyclyl, and heterocyclic ring backbone independently contains 1, 2, 3, or 4 heteroatoms selected from N, O, and S; the heterocyclic ring or heterocyclyl includes a saturated or partially unsaturated cyclic group; Each R 4 Independently selected from the following groups: hydrogen, deuterium, halogen, cyano, nitro, amino, C 1-6 Amine, hydroxyl, hydroxymethyl, carboxyl, mercapto, -S(O)2OH, C 1-6 alkylsulfonyl, R c C substituted or unsubstituted 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy groups, C 1-6 Alkylamine, C 3-8 cycloalkyl, C 3-8 Halogenated cycloalkyl, C 6-10 Aryl, 3-12 membered heterocyclic groups, -(CH2) p NH(CH2) p COOR 9 NR 10 (CH2) p -、-O(CH2) p NR 14 R 14’ ; R c selected from the group consisting of C 6-10 aryl, 5-7 membered heteroaryl, C 3-7 cycloalkyl, 4-10 membered heterocyclyl; the ring is selected from the group consisting of absent, 3-12 membered carbocyclo (including fused, bridged, and spiro rings), 3-12 membered heteromonocyclo, 7-20 membered heteropolycyclo (including fused, bridged, and spiro rings), 5-12 membered heteroaromatic ring; each R 5 and R 6 is independently selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxycarbonyl, C 1-6 haloalkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, C 3-8 halocycloalkyl, -(CH2) p -cyano, nitro, C 2-6 ester, amino, C 1-6 amine, hydroxyl, ketone carbonyl, C 1-6 alkylhydroxyl, carboxyl, thiol, C 1-6 alkylsulfonyl, -(CH2) p S(O)2R 13 , C6-C 10 aryl, 5-12 membered heteroaryl, 3-12 membered heterocyclyl, -C(O)(C 1-6 hydroxyalkyl), -(CH2) p NR 11 R 11’ , -(CH2) p NH(C 1-6 alkyl), -(CH2) p NH(CH2) p COOR 12 , -(CH2) p COOH; R 7 , R 8 , R 9 , R 10 , R 11 , R 11’ , R 12 , R 13 , R 14 and R 14’ are each independently selected from the group consisting of hydrogen, deuterium, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkoxycarbonyl, C 1-6 haloalkoxy, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, cyano, C 2-6 ester, nitro, amino, C 1-6 amine, hydroxyl, hydroxymethyl, carboxyl, C 6-10 aryl, 5-12 membered heteroaryl, 3-12 membered heterocyclyl; or R 7 , R 8 and the atom to which they are attached collectively form a 5-10 membered heterocyclyl; The halogen is F, Cl, Br, or I. The alkyl, alkoxy, alkenyl, alkynyl, cycloalkane, cycloalkyl, heterocyclic, heterocyclic, aryl, and heteroaryl groups are each independently substituted by 1-3 substituents selected from the group consisting of: halogen, C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 alkoxycarbonyl, C 1-6 Halogenated alkoxy groups, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 Cycloalkyl, cyano, nitro, amino, hydroxy, hydroxymethyl, carboxyl, mercapto, C 1-6 alkylsulfonyl, C 6-10 Aryl, 3-12 membered heterocyclic groups; wherein i is 0, 1, 2, 3, or 4.

2. The compound according to claim 1, and racemates, R-isomers, S-isomers, pharmaceutically acceptable salts thereof or mixtures thereof, characterized in that, the the ring is selected from the group consisting of saturated or partially saturated C 5-7 carbocyclic, C 6-10 aromatic, 5-6 membered heteroaromatic ring; Preferably, said the ring is selected from the group consisting of phenyl, furanyl, thienyl, pyrrolyl, thiazolyl, thiadiazolyl, oxadiazolyl, imidazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1,2,4,5-tetrazinyl.

3. The compound according to claim 1, racemates, R-isomers, S-isomers, pharmaceutically acceptable salts thereof or mixtures thereof, wherein The Ring is selected from the group consisting of 4-10 membered saturated carbocyclic, 4-10 membered saturated carbonspiro, 4-10 membered saturated or partially saturated heteromonocyclic, 7-10 membered heterofused ring, 7-10 membered heterospiro, 7-10 membered heterospiro, 5-9 membered heteroaromatic ring; Preferably, said the ring is selected from the group consisting of wherein i is 0, 1, 2, 3, or 4.

4. The compound according to claim 1, racemates, R-isomers, S-isomers, pharmaceutically acceptable salts thereof or mixtures thereof, wherein The selected from the group consisting of The structure of the compound is shown in Formula II:

5. The compound according to claim 1, racemates, R-isomers, S-isomers, pharmaceutically acceptable salts thereof or mixtures thereof, wherein, ###00003### The compound is a compound shown in the following table: wherein s, t, R 1 , R 2 , R 5 and R 6 are as defined in claim 1.

6. The compound according to claim 1, and racemates, R-isomers, S-isomers, pharmaceutically acceptable salts thereof or mixtures thereof, characterized in that, The pharmaceutical composition comprises one or more of the compound of Formula I, a pharmaceutically acceptable salt, racemate, R-isomer, S-isomer, or a mixture thereof of claim 1, and one or more of a pharmaceutically acceptable carrier, excipient, adjuvant, vehicle, and / or diluent.

7. A pharmaceutical composition, characterized by, A medicament for treating or preventing a disease associated with PL protease activity.

8. Use of a compound of formula I, a pharmaceutically acceptable salt, a racemate, an R-isomer, an S-isomer or a mixture thereof according to claim 1, or a pharmaceutical composition according to claim 7, characterized in that, The disease is a disease caused by a virus that has PL protease; preferably, the virus is selected from the group consisting of SARS-CoV-2, SARS-CoV, MERS-CoV, and HCoV-OC43, or a combination thereof.

9. Use according to claim 8, characterized in that, ​

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