Preparation method and application of benzimidazole skeleton-containing compound

Abstract

The invention relates to a preparation method and application of a compound containing a benzimidazole skeleton. Specifically, the invention discloses a compound as shown in a formula (I) and a pharmaceutical composition thereof, and the definition of each group or substituent group is described in the specification. The invention also provides a preparation method of the compound and an inhibition effect and application of the compound on a virus non-structural protein Nsp7 / 8 / 12 compound, and also discloses a brand-new heterocyclic skeleton compound, a targeted target spot has relatively high conservative property, and the compound can be used for broad-spectrum antivirus, and has a wide application prospect. Potential application prospects are realized in the field of medicines. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a preparation method and application of a compound containing a benzimidazole skeleton. Background Art

[0002] Since the outbreak of the novel coronavirus (SARS-CoV-2) at the end of 2019, it has spread rapidly and caused a major public health crisis worldwide. The pathogenic mechanism of the virus mainly depends on the efficient replication and transcription of its genome, which is mediated by the viral RNA-dependent RNA polymerase (RdRp) complex. The RdRp complex is composed of three non-structural proteins (nsps), nsp12, nsp7 and nsp8. Among them, nsp12 is the main catalytic subunit, while nsp7 and nsp8 provide structural stability and enhance the activity of RdRp by forming a multi-subunit complex.

[0003] In the RdRp complex, nsp8 is not only a co-factor of nsp12, but also directly participates in RNA chain extension and template replication through its unique structural and functional properties. Studies have shown that nsp8 can bind to nsp7 to form a "sliding clamp", which stabilizes the binding of the RNA template chain and the product chain and provides an additional active site for the RdRp complex, thereby achieving efficient synthesis of long-chain RNA. In addition, the high conservation of nsp8 makes it an ideal antiviral drug target. However, compared with the direct catalytic site of nsp12, the development of inhibitors for nsp8 is still in its early stages, and related structural studies and drug screening methods are still limited.

[0004] Drug development targeting nsp8 has significant advantages:

[0005] 1. High specificity: The function of nsp8 is irreplaceable in viral genome replication, and there is no corresponding functional homologous protein in host cells. Therefore, targeting nsp8 can minimize side effects on the host.

[0006] 2. Complex interference: nsp8 plays a key role in the assembly and stabilization of the RdRp complex. Targeting nsp8 can not only directly inhibit its activity, but also indirectly destroy the function of the nsp7 / 8 / 12 complex, achieving the effect of multi-target synergistic inhibition.

[0007] 3. Response to virus mutations: Since the sequence and structure of nsp8 are highly conserved in the coronavirus family, drugs targeting nsp8 are expected to be equally effective against coronavirus variants that may appear in the future.

[0008] Currently, most of the known small molecule inhibitors and antiviral drugs target the catalytic active center of nsp12, such as Remdesivir and Molnupiravir, but their efficacy is limited by viral mutations and drug tolerance. Therefore, the development of innovative inhibitors targeting nsp8 as a new approach to anti-SARS-CoV-2 drugs has important theoretical significance and application value.

[0009] Therefore, there is an urgent need to develop a new class of compounds in this field that can specifically bind to nsp8, block its interaction with nsp7 and nsp12, interfere with the assembly and function of the RdRp complex, and significantly inhibit the genome replication of SARS-CoV-2, thereby providing innovative strategies and candidate molecules for antiviral treatment. Summary of the invention

[0010] The main purpose of the present invention is to provide a benzimidazole heterocyclic derivative, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, and to screen out compounds with excellent antiviral properties in terms of effectiveness, safety and selectivity through structural innovation.

[0011] In a first aspect of the present invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof;

[0012]

[0013] in,

[0014] R1, R2, R3 and R4 are each independently selected from the group consisting of H, -NH2, halogen, -NO2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O and S, -C(=O )R6, -OC(=O)R6, -C(=O)OR6, -OR6, -SR6, -S(=O)R6, -S(=O)2R6, -S(=O)2N(R6)2, -N(R6)2, -C(=O)N(R6)2, -NR6-C(=O)R6, -NR6-C(=O)OR6, -NR6-S(=O)2-R6, -C1-C6 alkylene-N(R6)2, -C1-C6 alkylene-OR6, -C2-C6 alkenylene-OR6, -O-C1-C6 alkylene-N(R6)2;

[0015] Y is selected from the group consisting of C, N;

[0016] When Y is N, R3 does not exist;

[0017] R5 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S;

[0018] The substitution refers to substitution by one or more substituents selected from the group consisting of halogen, -NH2, -OH, C1-C6 alkoxy, -CN, -NO2, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy C1-C6 alkyl, C3-C10 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, C2-C6 acyl, -C(=O)OR6;

[0019] Wherein, the R6 is selected from the following group: H, guanidinyl, C1-C6 alkyl, saturated or partially unsaturated C3-C6 cycloalkyl, saturated or partially unsaturated C3-C10 heterocycloalkyl;

[0020] A is selected from the following group: absent, -OR 10 、-C1-C6 alkyl-R 10 、-N(R 10 )2, -C1-C6 alkylene-N(R 10 )2. -C(=O)-R 10、-NH-C(=O)-R 10 、-C(=O)-N(R 10 )2. -SO2-R 10 、-NH-SO2-R 10 、-SO2-N(R 10 )2;

[0021] R 10 Each is independently selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; or two R 10 The substituted or unsubstituted 5-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, or the substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution by a substituent selected from the group consisting of: -OH, -CN, -NH2, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(=O)-C1-C6 alkyl, -C(=O)-halogenated C1-C6 alkyl, -C(=O)-C2-C6 alkenyl, -C(=O)-C2-C6 alkynyl, -NH-C1-C C1-C6 alkyl, -NH-halo-C1-C6 alkyl, -NH-C2-C6 alkenyl, -NH-C2-C6 alkynyl, -N(C1-C6 alkyl)2, -N(halo-C1-C6 alkyl)2, -N(C2-C6 alkenyl)2, -N(C2-C6 alkynyl)2, -NH-C(=O)-C1-C6 alkyl, -NH-C(=O)-halo-C1-C6 alkyl, -COOH, -C(=O)-O-C1-C6 alkyl, -C(=O)-O-halo-C1-C6 alkyl, -C(=O)-O-C2-C6 alkenyl, C(=O)-O-C2-C6 alkynyl;

[0022] B is selected from the following groups: H, halogen, -NO2, C1-C6 alkyl, halo-substituted C1-C6 alkyl, saturated or partially unsaturated C3-C10 heterocycloalkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution by one or more substituents selected from the following groups: halogen, -NH2, -OH, oxo, -CN, -NO2, C1-C6 alkyl, halo-substituted C1-C6 alkyl, hydroxy C1-C6 alkyl, C1-C6 alkoxy, C3-C10 cycloalkyl, C3-C8 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, -COOH, C2-C6 acyl.

[0023] In another preferred embodiment, R1, R2, R3 and R4 are each independently selected from the following group: H, -NH2, halogen, -NO2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, substituted or unsubstituted C6-C10 aryl, containing 1-3 heteroatoms selected from N , substituted or unsubstituted 5-7 membered heteroaryl having O and S heteroatoms, -C(=O)R6, -OC(=O)R6, -C(=O)OR6, -OR6, -SR6, -S(=O)R6, -S(=O)2R6, -N(R6)2, -C(=O)N(R6)2, -NR6-C(=O)R6, -NR6-C(=O)OR6, -C1-C6 alkylene-N(R6)2, -C1-C6 alkylene-OR6, -C2-C6 alkenylene-OR6;

[0024] Y is C;

[0025] R5 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, -C(=O)R6, -OR6;

[0026] The substitution refers to substitution by one or more substituents selected from the group consisting of halogen, -NH2, -OH, C1-C6 alkoxy, -CN, -NO2, C1-C6 alkyl, C3-C10 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, -C(=O)OR6;

[0027] A is selected from the following group: absent, -OR 10 、-C1-C6 alkyl-R 10 、-N(R 10 )2. -C(=O)-R 10 、-NH-

[0028] C(=O)-R 10 、-C(=O)-N(R 10 )2. -SO2-R 10 、-SO2-N(R 10 )2;

[0029] R 10Each is independently selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; or two R 10 connected to form a substituted or unsubstituted 5-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution with a substituent selected from the following group: -OH, -CN, -NH2, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(=O)-C1-C6 alkyl, -C(=O)-halogenated C1-C6 alkyl, - C(=O)-C2-C6 alkenyl, -C(=O)-C2-C6 alkynyl, -NH-C1-C6 alkyl, -NH-haloC1-C6 alkyl, -NH-C2-C6 alkenyl, -NH-C2-C6 alkynyl, -N(C1-C6 alkyl)2, -NH-C(=O)-C1-C6 alkyl, -NH-C(=O)-haloC1-C6 alkyl, -COOH, -C(=O)-O-C1-C6 alkyl, -C(=O)-O-haloC1-C6 alkyl.

[0030] In another preferred embodiment, R1, R2, R3 and R4 are each independently selected from the following group: H, -NH2, halogen, -NO2, substituted or unsubstituted C1-C6 alkyl, -OR6;

[0031] R5 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, saturated or partially unsaturated substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S.

[0032] In another preferred embodiment, R5 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl;

[0033] In another preferred embodiment, the substitution refers to substitution by one or more substituents selected from the following group: -NH2, -OH, C1-C6 alkyl, -C(=O)OR6.

[0034] In another preferred embodiment, the substitution refers to substitution by one or more substituents selected from the following group: -NH2, -OH, -COOH.

[0035] A is selected from the following group: absent, -OR 10 、-C1-C6 alkyl-R 10 、-N(R 10 )2. -C(=O)-R 10 、-NH-C(=O)-R 10 、-C(=O)-N(R 10 )2. -SO2-R10 、-SO2-N(R 10 )2;

[0036] R 10 Each is independently selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; or two R 10 connected to form a substituted or unsubstituted 5-7 membered heterocycloalkyl group containing 1-3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 5-7 membered heteroaryl group containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution by a substituent selected from the following group: -OH, -CN, -NH2, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(=O)-C1-C6 alkyl, -C(=O)-halogenated C1-C6 alkyl, -C(=O)-C2-C6 alkenyl, -C(=O)-C2-C6 alkynyl, -NH-C(=O)-C1-C6 alkyl, -NH-C(=O)-halogenated C1-C6 alkyl, -COOH, -C(=O)-O-C1-C6 alkyl.

[0037] In another preferred embodiment, the compound of formula I is a compound of formula II,

[0038]

[0039] Wherein, R1, R2, R3, R4, R5 and B are as defined in the first aspect of the present invention;

[0040] Having a structure selected from the group consisting of:

[0041]

[0042] X is each independently selected from the following group:

[0043]

[0044] n is selected from the following group: 0, 1, 2, 3;

[0045] In another preferred embodiment, R1 and R2 are each independently selected from the following group: H, -OH, halogen, -NO2, C1-C6 alkyl, C1-C6 haloalkyl, -O-C1-C3 alkyl;

[0046] R3 and R4 are each independently selected from: H, -OH, halogen, -NO2, C1-C6 alkyl,

[0047] R5 is selected from the following group: H, -OH, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S; the substitution refers to substitution by one or more substituents selected from the following group: halogen, -NH2, -OH, C1-C6 alkoxy, -CN, -NO2, C1-C6 alkyl, halo C1-C6 alkyl, hydroxy C1-C6 alkyl, C3-C10 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, C2-C6 acyl, -C(=O)OR6;

[0048] In another preferred embodiment, R1 and R2 are each independently selected from the following group: H, -OH, halogen, -NO2, -CF3, -O-C1-C3 alkyl;

[0049] In another preferred embodiment, R3 and R4 are both H;

[0050] Having a structure selected from the group consisting of:

[0051]

[0052] X is selected from the following group:

[0053]

[0054] B is selected from the group consisting of H, halogen, -NO2, -CF3, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S. The substitution refers to substitution by one or more substituents selected from the group consisting of halogen, -NH2, -OH, oxo, -CN, -NO2, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy C1-C6 alkyl, C1-C6 alkoxy, C3-C10 cycloalkyl, C3-C8 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, -COOH, C2-C6 acyl.

[0055] In another preferred embodiment, the substitution refers to substitution by one or more substituents selected from the following group: halogen, -NH2, -OH, oxo, -NO2, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C3-C10 cycloalkyl, -COOH.

[0056] In another preferred embodiment, the substitution refers to substitution by one or more substituents selected from the group consisting of: -OH, -NH2, -COOH;

[0057] In another preferred embodiment, the compound is selected from the following group:

[0058]

[0059]

[0060]

[0061]

[0062] The second aspect of the present invention provides a method for preparing a compound of formula I, comprising the following steps:

[0063] In an inert solvent and at a certain temperature, compound I-1 reacts with compound I-2 to prepare a compound of formula I.

[0064]

[0065] The third aspect of the present invention provides a pharmaceutical composition comprising:

[0066] (1) The compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof; and

[0067] (2) A pharmaceutically acceptable carrier.

[0068] The fourth aspect of the present invention provides a use of a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, for preparing a drug or pharmaceutical composition for preventing and / or treating diseases caused by viral infection.

[0069] In another preferred embodiment, the disease is selected from the group consisting of coronavirus, calicivirus, and picornavirus infection.

[0070] In another preferred embodiment, the disease is caused by a viral infection selected from the group consisting of SARS-CoV-2 virus, SARS-CoV virus, MERS-CoV virus, and influenza virus.

[0071] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] Figure 1 The ability of the compounds to bind to nonstructural proteins is shown. DETAILED DESCRIPTION

[0073] After extensive and in-depth research and a large number of screening experiments, the inventors have developed a class of benzimidazole skeleton compounds. By substituting on the benzimidazole ring and the benzene ring connected thereto, the specific binding of the compound to nsp8 is increased, the activity of the RdRp complex is reduced, and RNA replication is inhibited.

[0074] the term

[0075] In the present invention, unless otherwise specified, the terms used have the general meanings well known to those skilled in the art.

[0076] In the present invention, the halogen is F, Cl, Br or I.

[0077] In the present invention, the term "C1-C6 alkyl" refers to a straight or branched alkyl group having 1 to 6 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl and n-hexyl, etc. "C1-C4 alkyl" and similar terms have similar definitions.

[0078] In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, etc. "C1-C3 alkoxy" and similar terms have similar definitions.

[0079] In the present invention, the term "C2-C8 alkenyl" refers to a straight or branched alkenyl group having 2 to 6 carbon atoms and containing at least one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl, etc. "C2-C6 alkenyl" and similar terms have similar definitions.

[0080] In the present invention, the term "C2-C8 alkynyl" refers to a straight or branched alkynyl group having 2 to 6 carbon atoms and containing at least one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl, etc. "C2-C6 alkynyl" and similar terms have similar definitions.

[0081] In the present invention, the term "C3-C8 cycloalkyl" represents a cyclic aliphatic hydrocarbon group consisting of 3 to 8 ring carbon atoms, and so on; it should be understood that the "cycloalkyl" described in the present invention includes not only monocyclic aliphatic hydrocarbon groups, but also cyclic, spirocyclic and bridged ring systems composed of multiple cyclic aliphatic hydrocarbons; the "cycloalkyl" described in the present invention includes not only aliphatic hydrocarbon groups with fully saturated carbon atoms, but also aliphatic hydrocarbon groups with unsaturated bonds in some carbon atoms; examples of the "cycloalkyl" described in the present invention include but are not limited to: When "cycloalkyl" is used as a substituent, the connection site with the main body of the molecule can occur at any position on the "cycloalkyl" that allows chemical bonds. "C3-C6 cycloalkyl" and other similar terms have similar definitions.

[0082] In the present invention, the term "aryl" refers to a monocyclic system and a bicyclic system composed of a specific number of carbon atoms and complying with Hückel's rule; it should be understood that when the "aryl" described in the present invention is a bicyclic system, it not only includes the case where all rings are aromatic rings, but also includes the case where only one ring is an aromatic ring and the other ring is a non-aromatic aliphatic ring.

[0083] In the present invention, the term "C6-C10 aryl" refers to a ring system having 6 to 10 carbon atoms, at least one of which is an aromatic ring; examples of the "aryl" described in the present invention include but are not limited to Etc.; when "aryl" is used as a substituent, the connection site with the main body of the molecule occurs on the aromatic ring.

[0084] In the present invention, the term "heterocycloalkyl" refers to a cyclic group that indicates a specific number of ring atoms, contains at least one ring heteroatom (N, O or S), is saturated or partially unsaturated, and is non-aromatic; it should be understood that the "heterocyclic group" described in the present invention includes not only monocyclic heterocyclic ring systems, but also polycyclic heterocyclic ring systems, such as cyclic, spirocyclic and bridged rings; when the "heterocyclic group" is a polycyclic system, at least one ring contains a ring heteroatom, and the other rings may contain ring heteroatoms or may be cycloalkyl; for example, the term "4-8 membered heterocycloalkyl" refers to a monocyclic or polycyclic system with 4 to 8 ring atoms, at least one of which is a heteroatom, saturated or partially unsaturated; the definitions of other similar terms are similar; preferably, the number of heteroatoms is 1 to 3. Including (but not limited to) the following groups: etc.; it should be understood that when a "heterocyclic group" is used as a substituent, the connection site with the main body of the molecule can occur at any position on the "heterocyclic group" that is allowed by a chemical bond.

[0085] In the present invention, the term "heteroaryl" refers to a cyclic group with a specific number of ring atoms, containing at least one ring heteroatom (N, O or S), and having aromaticity; unless otherwise specified, the "heteroaryl" described in the present invention includes not only a monocyclic heteroaromatic system, but also a polycyclic heteroaromatic system, such as a bicyclic heteroaromatic, a tricyclic heteroaromatic, and a tetracyclic heteroaromatic; when the "heteroaryl" described is a polycyclic heteroaromatic system, at least one ring is aromatic, the other rings may be aromatic or non-aromatic, and the heteroatom may be in an aromatic ring or in a non-aromatic ring; the polycyclic heteroaromatic system includes not only a paracyclic system, but also a bridged ring and a spirocyclic system. The term "5-7 membered heteroaryl" refers to a cyclic group with 5 to 7 ring atoms, at least one of which is a heteroatom and having aromaticity. The definitions of other similar terms are similar.

[0086] In the present invention, the term "oxo" refers to an oxygen atom connected to a carbon atom through a double bond to form a carbonyl (C=O) structure.

[0087] In the present invention, the term "halo" means substituted with halogen.

[0088] In the present invention, the term "optionally" means that when there are a series of candidate groups to choose from, some of them can be selected, or none of them can be selected.

[0089] The term "independently" used in the present invention means that when several substituents defined simultaneously are selected from the same series of candidate groups, they do not affect each other and may be the same or different.

[0090] In the present invention, the term "substituted" refers to one or more hydrogen atoms on a specific group being replaced by a specific substituent. The specific substituent is a substituent described above, or a substituent appearing in the embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituent may be the same or different at each position. It will be appreciated by those skilled in the art that the combinations of substituents contemplated by the present invention are those that are stable or chemically feasible.

[0091] In the present invention, the term "1-6" means 1, 2, 3, 4, 5 or 6, and other similar terms independently have similar meanings.

[0092] It should be understood that when a group is present in multiple different positions of a compound at the same time, its definition at each position is independent of each other and may be the same or different. That is, the term "selected from the following group:" has the same meaning as the term "each independently selected from the following group:".

[0093] Compound and preparation method thereof

[0094] For the first or second aspect of the present invention, the general formula (I) represents that the compound may contain one or more chiral centers, and there are enantiomers and diastereomers. The general formula (I) of the present invention represents that the compound may also contain many geometric isomers such as olefins, C=N double bonds, amides, etc. Unless otherwise specified, all chiralities (enantiomers, diastereomers, axial chiral isomers), racemates, cis geometric isomers, trans geometric isomers, cis and trans geometric isomer mixtures, rotational isomers and mixtures thereof described above are included in the present invention. A person skilled in the art may use a separation or synthesis method commonly used in the laboratory to separate or prepare the compound containing an asymmetric center in the present invention to obtain a single isomer. For example, for enantiomers, two enantiomers can be obtained by a general chiral resolution method or an asymmetric synthesis method. For diastereomers, they can be separated by methods such as fractional recrystallization or chromatographic separation, which does not destroy the novelty of the compounds of the present invention.

[0095] The compounds of the present invention have the structure shown in Formula I:

[0096]

[0097] Wherein, the definitions of each group are as described above;

[0098] In another preferred embodiment, in the compound, R1, R2, R3, R4, R5, R6, R 10 Any one of A, A1, B, n, X and Y is independently a corresponding group in the specific compound described in the present invention.

[0099] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a positively charged group on the compound of formula (I) and an anion, or a salt formed by a negatively charged group on the compound of formula (I) and a cation. Suitable anions include, but are not limited to, chloride, bromide, iodide, sulfate, nitrate, phosphate, citrate, methanesulfonate, trifluoroacetate, acetate, malate, toluenesulfonate, tartrate, fumarate, glutamate, glucuronide, lactate, glutarate or maleate, etc. Suitable cations include, but are not limited to, sodium, potassium, magnesium, calcium, ammonium, etc.

[0100] In another preferred embodiment, the pharmaceutically acceptable salt of the present invention refers to the salts formed by the compound represented by the general formula (I) and the following acid, such as but not limited to: hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, aminosulfonic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalene disulfonic acid, malonic acid, fumaric acid , propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid and isethionic acid, etc.; or a salt formed by the compound represented by the general formula (I) and an inorganic base, such as but not limited to sodium salt, potassium salt, calcium salt, aluminum salt or ammonium salt; or a salt formed by the compound represented by the general formula (I) and an organic base, such as but not limited to methylamine salt, ethylamine salt, ethanolamine salt, hydroxymethylaminomethane (TRIS) ammonium salt, etc.

[0101] The compound of the general formula (I) of the present invention or its pharmaceutically acceptable salt is separated out by distillation, crystallization or recrystallization from water or an organic solvent, and the compound may contain the solvent molecules used. In addition, different crystallization conditions may lead to different crystal forms of the compound. Therefore, the compound shown in the general formula (I) or its pharmaceutically acceptable salt containing different chemical doses of crystallization solvents and all crystal forms is within the scope of the present invention.

[0102] Replacing hydrogen atoms with deuterium atoms to change the physical and chemical properties of compounds has become a well-known structural modification method for those skilled in the art. Unless otherwise specified, the present invention intends to include the deuterated form of the compound represented by general formula (I) in the invention content.

[0103] The embodiments of the present invention specifically describe the preparation methods of the compounds of formula (I) of the present invention, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention belongs.

[0104] Typically, the raw materials and reagents used in the process for preparing the compounds of the present invention can be purchased through commercial channels unless otherwise specified.

[0105] Pharmaceutical compositions and methods of administration

[0106] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. The "safe and effective amount" means that the subject receiving the treatment at the dose is cured, improved, effectively prevented, or the incidence rate of the disease or side effects is significantly reduced compared with the subject not receiving the treatment at the dose; in addition, it also includes an effective dose for enhancing normal physiological functions. "Safety" means that the amount of the active ingredient is sufficient to significantly improve the condition without causing serious side effects.

[0107] In the pharmaceutical composition, the compound of the present invention is used as an active ingredient, and its weight accounts for 0.1-99.9% of the total weight of the pharmaceutical composition, and the rest are pharmaceutical excipients; the preferred ratio of the compound of the present invention to the excipients is: the compound of the present invention as an active ingredient accounts for more than 60% of the total weight, and the rest accounts for 0-40% of the total weight, and the amount of the rest is preferably 1-20%, and most preferably 1-10%. Usually, the pharmaceutical composition contains 1-2000 mg of active ingredient / dose, and more preferably, contains 10-200 mg of active ingredient / dose. Preferably, the "one dose" is one tablet.

[0108] The "pharmaceutical excipients" are pharmaceutically acceptable carriers, excipients, sustained-release agents, odorants, flavoring agents, etc. "Pharmaceutically acceptable carriers" refer to: one or more compatible solid or liquid fillers or gel substances, which 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 the active ingredients of the present invention and with each other without significantly reducing the efficacy of the active ingredients. Examples of pharmaceutically acceptable carriers include but are not limited to 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, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), etc.

[0109] In addition to the compound represented by general formula (I) as an active ingredient, the pharmaceutical composition may further contain one or more other therapeutic agents, wherein the "other therapeutic agents" are therapeutic agents for central nervous system diseases.

[0110] Adjuvants commonly used in the preparation of pharmaceutical compositions may also be included, and the "adjuvants" are flavoring agents, coloring agents, preservatives and antioxidants, such as vitamin E, vitamin C, BHT and BHA.

[0111] The compound or pharmaceutical composition of the present invention can be made into various dosage forms based on the conventional processes in the field of pharmaceutical preparations, such as tablets, capsules, powders, syrups, solutions, suspensions, sprays, creams, ointments, gels, transdermal patches, microneedles, etc., and can be present in suitable solid or liquid carriers or diluents. The pharmaceutical composition of the present invention can also be stored in suitable sterilizing apparatus for injection or instillation. From the standpoint of being easy to prepare and administer, the preferred pharmaceutical composition is a solid composition, especially tablets and solid-filled or liquid-filled capsules.

[0112] The compound or pharmaceutical composition of the present invention can be used clinically in mammals, including humans and animals. There is no particular limitation on the mode of administration, and representative modes of administration include but are not limited to oral administration, nasal inhalation, topical administration to the skin, intravenous injection, intramuscular injection, subcutaneous injection, etc. Preferably, the preferred route of administration of the compound or pharmaceutical composition of the present invention is oral administration.

[0113] 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 ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0114] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0115] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances, etc.

[0116] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0117] In addition to the active ingredients, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances.

[0118] Injectable preparations include, but are not limited to, sterile, injectable, aqueous, oily solutions, suspensions, emulsions, and the like. These preparations may also be formulated with suitable parenteral diluents, dispersants, wetting agents, suspending agents, and the like. Such injectable preparations may be sterilized by filtration in a filter that retains bacteria. These preparations may also be formulated with a bactericide that is dissolved or dispersed in an injectable medium or by other methods known in the art.

[0119] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0120] The treatment method of the present invention can be used alone or in combination with other treatment methods or therapeutic drugs (such as antiviral drugs).

[0121] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill of a skilled physician.

[0122] When used as a pharmaceutical preparation, the compound or pharmaceutical composition of the present invention can be used once a day or in divided doses. Regardless of the method of use, the optimal dose for an individual should be determined based on the specific treatment. Usually, it is started with a small dose and gradually increased until the most suitable dose is found.

[0123] Compared with the prior art, the present invention has the following main advantages:

[0124] 1. The compounds of the present invention can specifically bind to the nsp8 non-structural protein and inhibit RNA replication.

[0125] 2. The compounds of the present invention can significantly reduce the activity of the RdRp complex.

[0126] 3. The compounds of the present invention have significant antiviral effects.

[0127] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are generally performed under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0128] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0129] In the present invention, the structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR is determined using a Bruker AVANCE-400 nuclear magnetic spectrometer. LCMS is determined using a Waters 2695 liquid chromatography-mass spectrometer (MS model: Micromass ZQ).

[0130] The silica gel plate used in the thin layer chromatography (TLC) in the embodiment has a specification of 0.2mm±0.03mm. The thin layer chromatography separation and purification (prep-TLC) used for purifying compounds has a specification of 0.4mm~0.5mm; the column chromatography generally uses Yantai Huanghai silica gel 200~300 mesh silica gel as the carrier; the fully automatic medium pressure rapid purification instrument (Combi Flash Rf+UV-VIS) is used, and the separation column models are: Silica Flash Column 4g, 12g, 25g. The eluent system of the column chromatography and the development system of the thin layer chromatography include: A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as ammonia water or acetic acid can also be added for adjustment.

[0131] Preparation of intermediates

[0132] Example 1 Preparation of Intermediate S1 / S2

[0133]

[0134] Step 1: [(4-Fluoro-3-nitrophenyl)amino]methanoic acid-2-methylpropan-2-yl ester (S1)

[0135] The reactants 4-fluoro-3-nitroaniline (1.0 g, 6.4 mmol) and sodium hydroxide (256 mg, 6.4 mmol) were dissolved in a mixed solvent of 15 mL of dioxane and water (V:V = 4:1), and Boc anhydride (2.8 g, 12.8 mmol) was added dropwise under ice bath. After the addition was complete, the mixture was stirred at room temperature for 4 h. The reaction was complete after LC-MS detection. The reaction solution was dried and the crude product was purified by silica gel column chromatography (PE / EA = 3:1) to obtain a white solid product S1 (1.5 g, yield 91.5%).

[0136] 1 H NMR (400MHz, DMSO) δ9.85(s,1H),8.35(dd,J=7.0,2.8Hz,1H),7.75–7.67(m,1H),7.49(dd,J=11.2,9.1Hz,1H),1.49(s,9H). 13 C NMR (101MHz, DMSO) δ153.13,151.43,148.88,136.94,136.91,136.80,125.77,125.69,119.28,119.06,114.46,80.51,28.45.

[0137] Step 2: N-[4-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)-2-nitrophenyl]-DL-leucine-2-methylpropan-2-yl ester (S2)

[0138] The reactant [(4-fluoro-3-nitrophenyl)amino]methane acid-2-methylpropane-2-yl ester (1.0 g, 3.90 mmol) was dissolved in 8 mL of DMF, DL-leucine tert-butyl ester hydrochloride (1.1 g, 4.69 mmol) and triethylamine (1.59 g, 15.6 mmol) were added, and the reaction solution was heated at 90°C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, the organic phases were combined, and dried over anhydrous sodium sulfate. After flash purification (PE / EA=10:1), the product S2 (1.2 g, yield 73%) was obtained.

[0139] 1H NMR (500MHz, CDCl3) δ8.12–8.00(m,2H),7.63(s,1H),6.74(d,J=9.2Hz,1H),6.51(s,1H),4.09(q,J=7.3Hz,1H),1.84(dq ,J=13.6,6.8Hz,1H),1.78(td,J=6.9,1.9Hz,2H),1.53(s,9H),1.46(s,9H),1.03(d,J=6.5Hz,3H),0.96(d,J=6.4Hz,3H). 13 CNMR(126MHz, CDCl3)δ172.05,153.08,141.11,131.89,127.73,114.44,82.25,55.47,41.64,28.33,28.28,27.96,24.99,22.73,22.14.

[0140] Example 2 Preparation of Intermediate S3

[0141]

[0142] Step 1: 2-[2-(3,5-dichloro-2-hydroxyphenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (S2-1)

[0143] The reactant S2 (500 mg, 1.18 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (339 mg, 1.77 mmol), sodium dithionite (617 mg, 3.55 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and rinsed with cold water three times. The crude product was flash purified (PE / EA = 10:1) to obtain the product S2-1 (600 mg, yield 90%).

[0144] LC-MS: m / z=564.0 [M+H] +

[0145] Step 2: 2-[5-amino-2-(3,5-dichloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl]-4-methylpentanoic acid (S3)

[0146] The reactant S2-1 (600 mg, 1.06 mmol) was dissolved in 6 mL of dichloromethane, and 3 mL of trifluoroacetic acid was added dropwise under stirring. The mixture was reacted at room temperature for 4 h, and the reaction was complete after LC-MS monitoring. After HPLC purification, the solid product S3 (422 mg, yield 97%) was obtained.

[0147] LC-MS: m / z=408.1[M+H] +

[0148] 1 H NMR (500MHz, DMSO) δ9.50 (s, 1H), 7.84 (d, J = 2.6Hz, 1H), 7.71 (d, J = 8.8Hz, 1H), 7.52 (dd, J = 7.3, 2.4Hz, 2H), 7.21 (dd, J = 8.8, 2.1Hz,1H),4.89(dd,J=10.4,5.1Hz,1H),2.25–1.93(m,2H),1.10–0.99(m,1H),0.66(d,J=6.7Hz,3H),0.57(d,J=6.5Hz,3H). 13 C NMR (126MHz, DMSO) δ170.68,151.59,149.50,140.10,132.58,130.33,124 .09,123.70,118.94,117.86,115.15,58.11,38.60,24.65,22.80,21.16.

[0149] Preparation of compounds

[0150] Example 1

[0151]

[0152] Step 1: 2-Fluoro-1-nitrobenzene (200 mg, 1.42 mmol) was dissolved in 5 mL DMF, and DL-leucine (279 mg, 2.13 mmol) and anhydrous potassium carbonate (588 mg, 4.26 mmol) were added respectively. The reaction was carried out at room temperature for 4 h, and the reaction was complete after LCMS detection. The reaction solution was extracted with EA after adding water, and then dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation and purified by silica gel chromatography, using PE / EA=10:1 as the eluent, to obtain 301 mg of the product with a yield of 84%.

[0153] Step 2: N-(2-nitrophenyl)-DL-leucine (150 mg, 0.60 mmol) was dissolved in DMSO, and benzaldehyde (64 mg, 0.60 mmol) and sodium dithionite (313 mg, 1.8 mmol) were added. The reaction solution was heated at 90°C overnight, and then water was added to extract with EA, and then dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation and purified by silica gel chromatography, using PE / EA=5:1 as the eluent, to obtain 132 mg of product XD1, with a yield of 71%.

[0154] 1 H NMR (500MHz, CD3OD_SPE) δ7.96–7.83(m,5H),7.79(t,J=7.6Hz,2H),7.73–7.63(m,2H),5.50(dd,J=10.4,5.0Hz,1H),2.32(dd,J= 10.3, 4.5Hz, 1H), 2.20 (dd, J = 9.5, 5.1Hz, 1H), 1.14 (ddt, J = 16.2, 13.5, 5.7Hz, 1H), 0.74 (d, J = 6.7Hz, 3H), 0.59 (d, J = 6.5Hz, 3H). 13 C NMR(126MHz,CD3OD_SPE)δ169.78,151.67,133.02,132.62,131.47,129.7 0,129.67,126.49,126.14,123.31,115.15,114.09,58.58,48.12,47.95,4 7.91,47.86,47.81,47.78,47.72,47.68,47.60,47.55,47.54,47.49,47. 44,47.41,47.36,47.31,47.26,47.22,47.10,37.83,24.47,21.50,19.80.

[0155] Example 2

[0156]

[0157] The reaction steps are the same as those in Example 1. After the initial reactants are subjected to SNAr reaction, the fluorine atoms are replaced by amine reagents, and then the product containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite, and the target product is purified by HPLC.

[0158] 1H NMR (500MHz, CD3OD_SPE) δ7.96(d,J=7.4Hz,1H),7.91–7.84(m,1H),7.66(ddd,J=14.9,10.7,7.0Hz,4H),7.25–7.13(m,2H),5.26(dd,J=10.9,4.5 Hz,1H),2.45(ddd,J=14.9,11.0,4.2Hz,1H),2.14(ddd,J=14.5,9.8,4.6Hz,1H),1.23–1.06(m,1H),0.76(d,J=6.7Hz,3H),0.62(d,J=6.5Hz,3H). 13 C NMR (126MHz, CD3OD_SPE) δ170.05,156.71,135.11,131.23,126.46,125.93,120.24,116.34,115. 23,114.29,109.38,59.31,48.11,47.95,47.77,47.60,47.44,47.26,38.29,24.56,21.54,19.55.

[0159] Example 3

[0160]

[0161] The reaction steps are the same as those in Example 1. After the initial reactants are subjected to SNAr reaction, the fluorine atoms are replaced by amine reagents, and then the product containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite, and the target product is purified by HPLC.

[0162] 1 H NMR (500MHz, MeOD) δ7.99–7.85(m,2H),7.81(t,J=2.4Hz,2H),7.70–7.48(m,4H),5.16(dd,J=10.5,4.9Hz,2H),2.38(ddd,J=14.8,10.5,4.5H z,1H),2.18(ddd,J=14.4,9.4,4.9Hz,1H),1.47–1.29(m,1H),1.18(dtd,J=9.3,4.7,2.2Hz,1H),0.77(d,J=6.7Hz,3H),0.69(d,J=6.6Hz,4H). 13C NMR(126MHz,MeOD)δ170.10,151.53,147.87,133.46,131.82,129.64,125.67,125.47,124.96,123.14,115.81,114.7 7,114.52,58.89,48.49,48.14,47.96,47.79,47.62,47.45,47.28,47.11,38.36,24.53,21.56,21.54,19.87,19.86.

[0163] Example 4

[0164]

[0165] Step 1: 2-[2-(3,5-dichloro-2-hydroxyphenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (S2-1)

[0166] The reactant S2 (500 mg, 1.18 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (339 mg, 1.77 mmol), sodium dithionite (617 mg, 3.55 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and rinsed with cold water three times. The crude product was flash purified (PE / EA = 10:1) to obtain the product S2-1 (600 mg, yield 90%).

[0167] LC-MS: m / z=564.0 [M+H] +

[0168] Step 2: 2-[5-amino-2-(3,5-dichloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl]-4-methylpentanoic acid (S3)

[0169] The reactant S2-1 (600 mg, 1.06 mmol) was dissolved in 6 mL of dichloromethane, and 3 mL of trifluoroacetic acid was added dropwise under stirring. The mixture was reacted at room temperature for 4 h, and the reaction was complete after LC-MS monitoring. After HPLC purification, the solid product S3 (422 mg, yield 97%) was obtained.

[0170] LC-MS: m / z=408.1[M+H] +

[0171] 1H NMR (500MHz, DMSO) δ9.50 (s, 1H), 7.84 (d, J = 2.6Hz, 1H), 7.71 (d, J = 8.8Hz, 1H), 7.52 (dd, J = 7.3, 2.4Hz, 2H), 7.21 (dd, J = 8.8, 2.1Hz,1H),4.89(dd,J=10.4,5.1Hz,1H),2.25–1.93(m,2H),1.10–0.99(m,1H),0.66(d,J=6.7Hz,3H),0.57(d,J=6.5Hz,3H). 13 C NMR (126MHz, DMSO) δ170.68,151.59,149.50,140.10,132.58,130.33,124 .09,123.70,118.94,117.86,115.15,58.11,38.60,24.65,22.80,21.16.

[0172] Example 5

[0173]

[0174] Step 1: N-[4-(Acetylamino)-2-nitrophenyl]-DL-leucine-2-methylpropan-2-yl ester (5a)

[0175] The reactant N-(4-fluoro-3-nitrophenyl)acetamide (200 mg, 1.01 mmol) was dissolved in 4 mL of DMF, and DL-leucine tert-butyl ester hydrochloride (271 mg, 1.21 mmol) and triethylamine (412 mg, 4.04 mmol) were added. The reaction solution was heated at 90°C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The product 5a (271 mg, yield 73%) was obtained after flash purification (PE / EA=10:1).

[0176] LC-MS: m / z=366.1[M+H] +

[0177] 1H NMR (400MHz, CDCl3) δ8.40(s,1H),8.05(d,J=7.3Hz,1H),7.93–7.84(m,2H),6.68(d,J=9.1Hz,1H),4.16–4.06(m,1H),2. 14(s,3H),1.84(p,J=6.5Hz,1H),1.76(td,J=6.7,4.1Hz,2H),1.49(s,9H),1.03(d,J=6.3Hz,3H),0.95(d,J=6.2Hz,3H). 13 C NMR (101MHz, CDCl3) δ172.73,169.07,141.37,131.57,130.29,127.76,117.46,114.06,82.76,55.28,41.68,27.97,25.04,24.01,22.72,22.10.

[0178] Step 2: 2-[5-(Acetylamino)-2-(3,5-dichloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (5b)

[0179] The reactant 5a (177 mg, 0.48 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (111 mg, 0.58 mmol), sodium dithionite (253 mg, 1.45 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=3:1) to obtain the product 5b (194 mg, yield 79%).

[0180] LC-MS: m / z=506.1[M+H] +

[0181] 1 H NMR(400MHz, CDCl3) δ7.86(d,J=1.9Hz,1H),7.66–7.54(m,3H),7.51–7.41(m,2H),5.18(dd,J=9.7,5.8Hz,1H),2.24(s,3H),2.21–2.15( m,1H),1.99(ddd,J=14.5,8.8,5.8Hz,1H),1.50(s,9H),0.93(ddd,J=14.0,7.5,4.6Hz,1H),0.71(d,J=6.6Hz,3H),0.62(d,J=6.5Hz,3H). 13C NMR (101MHz, CDCl3) δ168.59,168.46,152.88,151.15,141.38,133.96,131.63,130.50,125.94,124. 19,123.89,117.87,115.56,113.09,111.09,83.78,59.07,38.33,27.93,24.62,24.50,22.55,21.22.

[0182] Step 3: 2-[5-(Acetylamino)-2-(3,5-dichloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl]-4-methylpentanoic acid (XD5)

[0183] The reactant 5b (150 mg, 0.30 mmol) was dissolved in 6 mL of dichloromethane, and 3 mL of trifluoroacetic acid was added dropwise under stirring. The mixture was reacted at room temperature for 4 h, and the reaction was completed after LC-MS monitoring. After HPLC purification, the solid product XD5 (115 mg, yield 87%) was obtained.

[0184] LC-MS: m / z=450.1[M+H] +

[0185] 1 H NMR (400MHz, DMSO) δ10.34(s,1H),8.30(d,J=1.9Hz,1H),7.90(d,J=2.6Hz,1H),7.7 8(d,J=9.0Hz,1H),7.66(d,J=2.6Hz,1H),7.55(dd,J=9.0,1.9Hz,1H),4.97(dd,J=1 0.1,5.2Hz,1H),2.22(ddd,J=14.6,10.1,4.7Hz,1H),2.11(s,3H),2.04(ddd,J=14. 4,7.4,3.3Hz,1H),1.18–1.06(m,1H),0.68(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H). 13 CNMR(101MHz,DMSO)δ170.27,169.16,151.93,148.03,137.20,136.24,133.35,130.29,128.2 7,124.14,123.93,118.17,116.59,115.08,106.17,58.57,38.55,24.61,24.44,22.76,21.20.

[0186] Example 6

[0187]

[0188] Step 1: N-{4-[(methylamino)carbonyl]-2-nitrophenyl}-DL-leucine-2-methylpropan-2-yl ester (6a)

[0189] The reactant 4-fluoro-N-methyl-3-nitrobenzamide (200 mg, 1.01 mmol) was dissolved in 4 mL of DMF, DL-leucine tert-butyl ester hydrochloride (271 mg, 1.21 mmol) and triethylamine (412 mg, 4.04 mmol) were added, and the reaction solution was heated at 90°C overnight. The reaction solution was then extracted three times with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. After flash purification (PE / EA=10:1), the product 6a (255 mg, yield 69%) was obtained.

[0190] LC-MS: m / z=366.1[M+H] +

[0191] 1 H NMR (400MHz, CDCl3) δ8.59(d,J=2.2Hz,1H),8.43(d,J=7.4Hz,1H),7.99(dd,J=9.0,2.2Hz,1H),6.81(d,J=9.0Hz,1H),6.50(d,J=5.3Hz,1H),4.12 (d,J=7.1Hz,1H),3.00(d,J=4.8Hz,3H),1.82–1.79(m,2H),1.68–1.65(m ,1H),1.46(d,J=3.0Hz,9H),1.04(d,J=6.0Hz,3H),0.96(d,J=4.4Hz,3H). 13 CNMR(101MHz, CDCl3)δ176.00,171.24,171.18,165.99,145.99,135.19,131.42,125.40,122.28,114.05,82.67,80.77, 60.40,55.32,53.46,44.28,41.47,28.04,27.94,26.90,26.81,25.00,24.86,22.92,22.66,22.18,21.97,21.03,14.18.

[0192] Step 2: 2-[2-(3,5-dichloro-2-hydroxyphenyl)-5-[(methylamino)carbonyl]benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (6b)

[0193] The reactant 6a (184 mg, 0.50 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (116 mg, 0.60 mmol), sodium dithionite (263 mg, 1.51 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=3:1) to obtain the product 6b (117 mg, yield 46%).

[0194] LC-MS: m / z=506.1[M+H] +

[0195] 1 H NMR (400MHz, CDCl3) δ7.74(s,1H),7.63–7.56(m,1H),7.49(d,J=8.6Hz,1H),7.42(d,J=2 .6Hz,1H),7.29(d,J=2.6Hz,1H),6.82(d,J=6.0Hz,1H),4.80(d,J=11.6Hz,1H),3.08(d, J=4.7Hz,3H),2.26(ddd,J=14.9,10.9,4.4Hz,1H),1.81(ddt,J=16.2,11.8,5.9Hz,1H), 1.61(s,9H),0.78(dt,J=13.3,5.8Hz,1H),0.62(d,J=6.6Hz,3H),0.54(d,J=6.5Hz,3H). 13 C NMR (101MHz, CDCl3) δ169.71,168.43,151.69,151.60,140.14,134.41,132.07,130.08,127. 58,125.44,123.27,117.49,112.99,83.80,58.92,38.79,28.09,26.89,24.68,22.54,20.89.

[0196] Step 3: 2-[2-(3,5-dichloro-2-hydroxyphenyl)-5-[(methylamino)carbonyl]benzo[d]imidazol-1-yl]-4-methylpentanoic acid (XD6)

[0197] The reactant 6b (117 mg, 0.23 mmol) was dissolved in 6 mL of dichloromethane, and 3 mL of trifluoroacetic acid was added dropwise under stirring. The mixture was reacted at room temperature for 4 h, and the reaction was completed after LC-MS monitoring. After HPLC purification, the solid product XD6 (76 mg, yield 73%) was obtained.

[0198] LC-MS: m / z=450.1[M+H] +

[0199] 1 H NMR (400MHz, DMSO) δ9.01 (s, 1H), 8.60 (q, J = 4.5Hz, 1H), 8.30 (s, 1H), 7.96–7.88 (m, 1H), 7.85–7.75 (m, 2H), 7.56 (d, J = 2.6Hz, 1H), 4. 95(dd,J=10.1,5.2Hz,1H),2.85(d,J=4.2Hz,3H),2.30–1.95(m,2H),1.16–1.03(m,1H),0.66(d,J=6.6Hz,3H),0.59(d,J=6.5Hz,3H). 13 C NMR (101MHz, DMSO) δ170.78,166.99,151.61,150.74,139.85,135.27,132.49,130.43,130.26 ,124.13,123.71,123.54,119.22,117.78,113.85,58.10,38.64,26.83,24.69,22.74,21.20.

[0200] Example 7

[0201]

[0202] The reaction steps are the same as those in Example 1. After the initial reactants are subjected to SNAr reaction, the fluorine atoms are replaced by amine reagents, and then the product containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite, and the target product is purified by HPLC.

[0203] LC-MS: m / z=436.1[M+H] +

[0204] Example 8

[0205]

[0206] The reaction steps are the same as those in Example 1. After the initial reactants are subjected to SNAr reaction, the fluorine atoms are replaced by amine reagents, and then the product containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite, and the target product is purified by HPLC.

[0207] 1H NMR(500MHz,MeOD)δ8.26(d,J=1.5Hz,1H),7.91–7.83(m,2H),7.72(d,J=2.5Hz,1H),7.52(d,J=2.6Hz,1H),5.02(dd, J=10.5,4.9Hz,1H),2.59(s,3H),2.44–2.02(m,2H),1.18–1.07(m,1H),0.76(d,J=6.7Hz,3H),0.68(d,J=6.5Hz,3H). 13 C NMR(126MHz,MeOD)δ136.05,131.98,129.84,124.69,121.73,118.46,114.13,58.07,48.11,48.04,47.94, 47.88,47.81,47.77,47.76,47.67,47.60,47.49,47.43,47.26,47.09,38.63,27.93,24.60,21.51,19.89.

[0208] Example 9

[0209]

[0210] XD4 (60 mg, 0.15 mmol) was dissolved in 2 mL of dichloromethane, triethylamine (44.6 mg, 0.44 mmol) was added to the reaction solution, and stirred for 10 min under ice bath. Chloroacetyl chloride (17.4 mg, 0.15 mmol) in dichloromethane was slowly added dropwise at 0°C. After the reaction was completed by LCMS, 61 mg of the final product XD9 was obtained by HPLC purification with a yield of 86%.

[0211] 1 H NMR (500MHz, DMSO) δ10.53(s,1H),8.15(d,J=2.0Hz,1H),7.85(d,J=2.5Hz,1H),7.68(d,J=8.9Hz,1H),7.54(d,J=2.6Hz,1H),7.48(dd,J= 9.0,2.0Hz,1H),4.91(dd,J=10.2,5.2Hz,1H),4.31(s,2H),2.21–1.92(m,2H),1.03(s,1H),0.66(d,J=6.6Hz,3H),0.56(d,J=6.5Hz,3H). 13C NMR (126MHz, DMSO) δ170.81,165.21,151.65,149.56,135.06,132.45,130.12,129.94,12 4.02,123.67,118.93,117.47,114.35,108.74,58.03,44.07,38.57,24.64,22.84,21.24.

[0212] Example 10

[0213]

[0214] Step 1: [(3-Fluoro-4-nitrophenyl)amino]methanoic acid-2-methylpropan-2-yl ester (10a)

[0215] The reactants 3-fluoro-4-nitroaniline (200 mg, 1.28 mmol) and sodium hydroxide (52 mg, 1.31 mmol) were dissolved in 5 mL of a mixed solvent of dioxane and water (V:V = 4:1), and Boc anhydride (560 mg, 2.56 mmol) was added dropwise under an ice bath. After the addition was complete, the mixture was stirred at room temperature for 4 h. The reaction was complete when detected by LC-MS. The reaction solution was dried and the crude product was purified by silica gel column chromatography (PE / EA = 10:1) to obtain a white solid product 10a (311 mg, yield 95%).

[0216] 1 H NMR (500MHz, CDCl3) δ7.63 (dd, J=13.5, 2.4Hz, 1H), 7.09 (ddd, J=9.1, 2.4, 1.1Hz, 1H), 6.95 (s, 1H), 1.55 (s, 9H). 13 C NMR (126MHz, CDCl3) δ151.53,145.58,127.36,112.86,112.83,106.73,106.52,84.41,82.47,28.16.

[0217] Step 2: N-[5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)-2-nitrophenyl]-DL-leucine-2-methylpropan-2-yl ester (10b)

[0218] The reactant 10a (200 mg, 0.78 mmol) was dissolved in 4 mL of DMF, and DL-leucine tert-butyl ester hydrochloride (210 mg, 0.94 mmol) and triethylamine (318 mg, 3.12 mmol) were added. The reaction solution was heated at 90°C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The product 10b (190 mg, yield 58%) was obtained after flash purification (PE / EA=10:1).

[0219] LC-MS: m / z = 424.2 [M+H] +

[0220] 1 H NMR (400MHz, CDCl3) δ8.46(d,J=7.1Hz,1H),8.09(d,J=9.3Hz,1H),6.92(s,1H),6.37(dd,J=9.3,2.2Hz,1H),4.17–4.11( m,1H),1.86(d,J=2.5Hz,1H),1.79(t,J=6.7Hz,2H),1.52(s,9H),1.49(s,9H),1.04(d,J=6.2Hz,3H),0.97–0.95(m,3H). 13 C NMR (101MHz, CDCl3) δ171.95,151.70,146.19,145.85,128.41,127.63,106.96,100 .06,82.25,81.48,55.23,41.47,31.58,28.17,27.87,25.03,22.76,22.64,22.18.

[0221] Step 3: 2-methylpropan-2-yl {[2-(3,5-dichloro-2-hydroxyphenyl)-3-(3-methylbutyl)benzo[d]imidazol-5-yl]amino}methanoate (10c)

[0222] The reactant 10b (150 mg, 0.35 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (81 mg, 0.43 mmol), sodium dithionite (185 mg, 1.06 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=3:1) to obtain the product 10c (105 mg, yield 52%).

[0223] LC-MS: m / z=564.2[M+H]+

[0224] 1 H NMR (400MHz, CDCl3) δ7.82(d,J=2.0Hz,1H),7.61(dd,J=5.6,3.1Hz,2H),7.46(d,J=2.5Hz,1H),7.25–7.18(m,1H),6.77(s,1H),5.21 (dd,J=9.7,5.6Hz,1H),1.57(s,9H),1.52(s,9H),1.06–0.95(m,2H),0.91–0.86(m,1H),0.72(d,J=6.6Hz,3H),0.64(d,J=6.5Hz,3H). 13 C NMR (101MHz, CDCl3) δ168.51,152.80,150.16,137.08,135.19,133.99,131.29,125.83,124. 02,123.74,119.62,116.13,115.71,83.77,59.00,38.12,28.39,27.92,24.73,22.60,21.26.

[0225] Step 4: 2-{6-[(2-chloroacetyl)amino]-2-(3,5-dichloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD10)

[0226] The reactant 10c (105 mg, 0.19 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (73 mg, 0.74 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (21 mg, 0.19 mmol) was added dropwise while stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was complete as monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD10 (63 mg, yield 70%).

[0227] LC-MS: m / z=484.1[M+H] +

[0228] 1H NMR (400MHz, DMSO) δ11.12(s,1H),8.27(d,J=1.9Hz,1H),7.86(d,J=2.6Hz,1H),7.79(d,J=8.8Hz,1H),7.66(dd,J=8.9,1.8Hz,1H),7.59(d,J=2.6Hz ,1H),4.93(dd,J=9.8,5.3Hz,1H),4.39(s,2H),2.55(s,3H),2.29–1.93(m ,2H),1.11(q,J=9.3Hz,1H),0.67(d,J=6.6Hz,3H),0.57(d,J=6.4Hz,3H). 13 C NMR(101MHz,DMSO)δ170.35,165.41,151.78,148.59,136.09,132.84,132.79,130.27,124.0 8,123.82,117.98,117.72,117.51,104.31,58.13,44.08,40.82,38.37,24.66,22.80,21.26.

[0229] Embodiment 11

[0230]

[0231] Step 1: 4-Fluoro-3-nitrobenzoic acid (400 mg, 2.16 mmol) was dissolved in 6 mL of DMF, and DL-leucine tert-butyl ester (606 mg, 3.24 mmol) and anhydrous potassium carbonate (894 mg, 6.48 mmol) were added respectively. The reaction was carried out at room temperature for 4 hours, and the reaction was complete after LCMS detection. The reaction solution was extracted with EA after adding water, and then dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation and purified by silica gel chromatography, using PE / EA=3:1 as the eluent, to obtain 522 mg of the product with a yield of 69%.

[0232] Step 2: N-(4-carboxyl-2-nitrophenyl)-DL-leucine-2-methylpropane-2-yl ester (500 mg, 1.42 mmol) was dissolved in DMF, and piperazine-1-carboxylic acid-2-methylpropane-2-yl ester (264 mg, 1.42 mmol), HATU (809 mg, 2.13 mmol), and DIPEA (550 mg, 4.26 mmol) were added respectively. Stir at room temperature until the reaction was completed by LCMS detection. The reaction solution was added with water and extracted with EA, and then dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation and purified by silica gel chromatography, using PE / EA=5:1 as the eluent to obtain 566 mg of the product with a yield of 77%.

[0233] 1 H NMR (500MHz, CDCl3) δ8.40(d,J=7.4Hz,1H),8.32(d,J=2.1Hz,1H),7.59(dd,J=8.9,2.1Hz,1H),6.82(d,J=8.8Hz,1H),4.23–4.01(m,2H),3.62(s,3H ),3.49(t,J=5.3Hz,4H),2.04(dd,J=17.5,1.8Hz,3H),1.91–1.72(m,4H), 1.49(d,J=6.5Hz,20H), 1.05(d,J=6.1Hz,3H), 0.97(dd,J=6.3,4.0Hz,5H).

[0234] Step 3: 4-({4-[(3-methylbutyl)amino]-3-nitrophenyl}carbonyl)piperazine-1-carboxylic acid-2-methylprop-2-yl ester (150 mg, 0.29 mmol) was dissolved in DMSO, and 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (55 mg, 0.29 mmol) and sodium dithionite (151 mg, 0.87 mmol) were added respectively. The reaction solution was heated at 90°C for overnight reaction, water was added and extracted with EA, and then dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation and purified by silica gel chromatography, using PE / EA=5:1 as the eluent to obtain 125 mg of the product with a yield of 65%.

[0235] Step 4: 4-{[2-(3,5-dichloro-2-hydroxyphenyl)-1-(3-methylbutyl)benzo[d]imidazol-5-yl]carbonyl}piperazine-1-carboxylic acid-2-methylpropane-2-yl ester (125 mg, 0.19 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added and stirred at room temperature for 4 h. The reaction solution was spin-dried and the crude product was directly used for subsequent reactions without post-treatment. The crude product was redissolved in DCM, NEt3 (96 mg, 0.95 mmol) was added and stirred at 0°C, and chloroacetyl chloride (21.3 mg, 0.19 mmol) was added dropwise to the reaction solution. The reaction solution was stirred until the reaction was complete as monitored by LCMS. 44 mg of the target product was purified by HPLC with a yield of 40%.

[0236] 1H NMR (500MHz, DMSO) δ7.81(d,J=2.6Hz,2H),7.69(d,J=8.4Hz,1H),7.45(d,J=2.6Hz,1H),7.39(dd,J=8.4,1.6Hz,1H),4.88(dd,J=10.1 ,5.3Hz,1H),4.42(s,2H),3.55(s,8H),2.18–1.97(m,2H),1.06(dt,J=14.0,6.7Hz,1H),0.66(d,J=6.6Hz,3H),0.58(d,J=6.6Hz,3H). 13 C NMR (126MHz, DMSO) δ171.11,170.00,165.31,151.45,151.13,141.90,134.84,131.92,130.34,130.17,123.99,1 23.49,122.88,120.78,118.66,117.62,115.29,113.69,57.69,46.09,42.43,38.69,24.71,22.84,21.28,9.01.

[0237] Example 12

[0238]

[0239] The reaction steps are the same as those in Example 11. The fluorine atom of the initial reactant is replaced by an amine reagent after the SNAr reaction, and then an amide condensation is carried out with a Boc-protected diamine reagent. The intermediate containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite. The intermediate is deprotected under the action of trifluoroacetic acid and then purified by HPLC to obtain the target product.

[0240] 1 H NMR (500MHz, DMSO) δ8.61(t,J=5.5Hz,1H),8.38(t,J=5.7Hz,1H),8.26(d,J=1.6 Hz,1H),7.88–7.80(m,2H),7.71(d,J=8.6Hz,1H),7.48(d,J=2.6Hz,1H),4.90(d d,J=10.3,5.1Hz,1H),4.08(s,2H),3.43–3.37(m,2H),3.32(q,J=6.2Hz,2H),2. 26–1.93(m,2H),1.08–0.97(m,1H),0.65(d,J=6.7Hz,3H),0.57(d,J=6.5Hz,3H). 13C NMR (126MHz, DMSO) δ171.03,167.00,166.69,158.90,158.62,151.46,151.17,141.51,135.86,132.08,130.2 2,129.70,124.06,123.58,123.20,120.41,118.62,113.41,57.80,43.15,39.33,38.62,24.69,22.85,21.23.

[0241] Embodiment 13

[0242]

[0243] Step 1: {[4-(Methylamino)-3-nitrophenyl]amino}methanoic acid-2-methylpropan-2-yl ester (13a)

[0244] The reactant S1 (200 mg, 0.78 mmol) was dissolved in 4 mL DMF, and methylamine hydrochloride (63 mg, 0.94 mmol) and triethylamine (318 mg, 3.12 mmol) were added. The reaction solution was heated at 110°C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. After flash purification (PE / EA=10:1), the product 13a (196 mg, yield 94%) was obtained.

[0245] LC-MS: m / z=268.1[M+H] +

[0246] 1 H NMR (400MHz, CDCl3) δ8.11(d,J=2.6Hz,1H),7.92(d,J=6.7Hz,1H),6.82(d,J=9.2Hz,1H),6.44(s,1H),3.03(d,J=5.1Hz,3H),1.53(s,9H). 13 CNMR(101MHz, CDCl3)δ153.21,143.29,131.07,129.79,126.87,113.89,29.85,28.33.

[0247] Step 2: 2-methylpropan-2-yl {[2-(3,5-dichloro-2-hydroxyphenyl)-1-methylbenzo[d]imidazol-5-yl]amino}methanoate (13b)

[0248] The reactant 13a (196 mg, 0.73 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (166 mg, 0.88 mmol), sodium dithionite (383 mg, 2.2 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=3:1) to obtain the product 13b (166 mg, yield 56%).

[0249] LC-MS: m / z=407.8[M+H] +

[0250] 1 H NMR (400MHz, DMSO) δ9.42(s,1H),7.92(s,1H),7.84(d,J=2.5Hz,1H),7.72(d,J=2.5H z,1H),7.61(d,J=8.8Hz,1H),7.43(dd,J=8.8,2.0Hz,1H),3.98(s,3H),1.51(s,9H). 13 C NMR (101MHz, DMSO) δ153.45,153.19,149.77,140.10,135.95,131.74,131.08,127.16,122.91,122.83,117.21,111.13,79.43,33.02,28.67.

[0251] Step 3: 2-Chloro-N-[2-(3,5-dichloro-2-hydroxyphenyl)-1-methylbenzo[d]imidazol-5-yl]acetamide (XD13)

[0252] The reactant 13b (166 mg, 0.41 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring, and the reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (165 mg, 1.63 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (46 mg, 0.41 mmol) was added dropwise while stirring under ice bath, and the reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD13 (79 mg, yield 50%).

[0253] LC-MS: m / z=383.8[M+H] +

[0254] 1H NMR (400MHz, DMSO) δ10.62(s,1H),8.21(d,J=1.9Hz,1H),7.88–7.74(m,3H),7.58(dd,J=8.8,1.9Hz,1H),4.33(s,2H),3.93(s,3H). 13 C NMR (101MHz, DMSO) δ165.27,152.61,148.86,136.83,135.88,132.36,131. 53,128.72,123.54,123.41,117.87,116.17,112.46,107.26,44.09,32.88.

[0255] Embodiment 14

[0256]

[0257] Step 1: {[4-(ethylamino)-3-nitrophenyl]amino}methanoic acid-2-methylpropan-2-yl ester (14a)

[0258] The reactant S1 (200 mg, 0.78 mmol) was dissolved in 4 mL DMF, and ethylamine (53 mg, 1.17 mmol) and triethylamine (318 mg, 3.12 mmol) were added. The reaction solution was heated at 90°C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The product 14a (200 mg, yield 91%) was obtained after flash purification (PE / EA=10:1).

[0259] LC-MS: m / z=282.1[M+H] +

[0260] 1 H NMR(500MHz, CDCl3)δ8.09(d,J=2.7Hz,1H),7.88–7.82(m,1H),6.81(d,J=9.2Hz, 1H), 6.49 (s, 1H), 3.34 (qd, J = 7.2, 5.1Hz, 2H), 1.52 (s, 9H), 1.37 (t, J = 7.2Hz, 3H). 13 C NMR (126MHz, CDCl3) δ153.27,142.46,130.84,129.82,126.76,114.28,37.80,28.33,14.42.

[0261] Step 2: 2-methylpropan-2-yl {[2-(3,5-dichloro-2-hydroxyphenyl)-1-ethylbenzo[d]imidazol-5-yl]amino}methanoate (14b)

[0262] The reactant 14a (200 mg, 0.71 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (161 mg, 0.85 mmol), sodium dithionite (372 mg, 2.1 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=3:1) to obtain the product 14b (180 mg, yield 60%).

[0263] LC-MS: m / z=422.0 [M+H] +

[0264] 1 H NMR (400MHz, CDCl3) δ7.74(d,J=2.0Hz,1H),7.50(d,J=2.4Hz,1H),7.45(d,J=2.4Hz,1H),7.39(d,J=8. 1Hz, 1H), 7.33 (d, J = 8.7Hz, 1H), 6.65 (s, 1H), 4.45 (q, J = 7.3Hz, 2H), 1.63 (t, J = 7.3Hz, 3H), 1.57 (s, 9H). 13 C NMR (101MHz, CDCl3) δ154.13,153.14,149.38,139.94,134.65,131.14,131.10,124 .12,124.00,122.92,117.09,114.75,109.83,108.95,80.68,40.79,28.41,15.10.

[0265] Step 3: 2-Chloro-N-[2-(3,5-dichloro-2-hydroxyphenyl)-1-ethylbenzo[d]imidazol-5-yl]acetamide (XD14)

[0266] The reactant 14b (180 mg, 0.43 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (173 mg, 1.71 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (57 mg, 0.51 mmol) was added dropwise under stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD14 (120 mg, yield 71%).

[0267] LC-MS: m / z=398.0[M+H] +

[0268] 1 H NMR (400MHz, DMSO) δ10.84(s,1H),8.33(d,J=1.9Hz,1H),8.00(d,J=9.0Hz,1H),7.89(d,J=2.6Hz,1 H),7.76(d,J=2.6Hz,1H),7.68(dd,J=9.0,1.9Hz,1H),4.35(d,J=8.4Hz,4H),1.38(t,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO)δ165.56,152.16,146.85,137.05,134.00,133.48,129.87, 129.17,124.02,123.87,118.70,115.08,113.74,105.61,44.03,41.41,14.65.

[0269] Embodiment 15

[0270]

[0271] Step 1: ({4-[(3-methylbutyl)amino]-3-nitrophenyl}amino)methanoic acid-2-methylpropan-2-yl ester (15a)

[0272] The reactant S1 (200 mg, 0.78 mmol) was dissolved in 4 mL DMF, 3-methylbutan-1-amine (82 mg, 0.94 mmol) and triethylamine (239 mg, 2.34 mmol) were added, and the reaction solution was heated at 90°C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, the organic phases were combined and dried over anhydrous sodium sulfate. After flash purification (PE / EA=20:1), the product 15a (186 mg, yield 74%) was obtained.

[0273] LC-MS: m / z=324.1[M+H] +

[0274] 1H NMR (500MHz, CDCl3) δ8.09(d,J=2.6Hz,1H),7.91(t,J=4.9Hz,1H),6.83(d,J=9.2Hz,1H),6.45(s,1H),3.31(td,J =7.3,5.1Hz,2H),1.86–1.70(m,2H),1.53(s,9H),1.20–1.11(m,1H),0.99(d,J=3.6Hz,3H),0.98(d,J=3.6Hz,3H). 13 C NMR (126MHz, CDCl3) δ153.23,142.61,130.86,129.80,126.69,114.31,41.37,41.35,37.87,31.59,28.32,25.95,22.47.

[0275] Step 2: 2-methylpropan-2-yl {[2-(3,5-dichloro-2-hydroxyphenyl)-1-(3-methylbutyl)benzo[d]imidazol-5-yl]amino}methanoate (15b)

[0276] The reactant 15a (186 mg, 0.27 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (131 mg, 0.69 mmol), sodium dithionite (301 mg, 1.73 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=5:1) to obtain the product 15b (177 mg, yield 66%).

[0277] LC-MS: m / z=464.1[M+H] +

[0278] 1 H NMR (400MHz, CDCl3) δ7.71(d,J=1.9Hz,1H),7.48(d,J=2.4Hz,1H),7.42(d,J=2.4Hz,1H),7.32(dd,J=8.6,1.8Hz,1H),7.29–7.2 6(m,1H),6.66(s,1H),4.37–4.28(m,2H),1.84(dd,J=6.8,4.6Hz,2H),1.57(s,9H),1.28(t,J=7.1Hz,1H),1.09(d,J=6.2Hz,6H). 13C NMR (101MHz, CDCl3) δ154.08,153.11,149.31,139.73,134.59,131.25,131.01,124.19,12 3.91,122.87,117.00,114.67,109.83,108.81,80.61,44.42,38.34,28.41,26.32,22.36.

[0279] Step 3: 2-Chloro-N-[2-(3,5-dichloro-2-hydroxyphenyl)-1-(3-methylbutyl)benzo[d]imidazol-5-yl]acetamide (XD15)

[0280] The reactant 15b (177 mg, 0.38 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring, and the reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (155 mg, 1.53 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (43 mg, 0.38 mmol) was added dropwise under stirring under ice bath, and the reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD15 (115 mg, yield 69%).

[0281] LC-MS: m / z=440.1[M+H] +

[0282] 1 H NMR (400MHz, DMSO) δ10.79(s,1H),8.30(d,J=1.9Hz,1H),7.92(d,J=9.0Hz,1H),7.86(d,J=2.5Hz,1H),7.75(d,J=2.6Hz,1H ),7.64(dd,J=8.9,1.9Hz,1H),4.33(d,J=12.5Hz,4H),1.71–1.60(m,2H),1.53(hept,J=6.6Hz,1H),0.80(d,J=6.6Hz,6H). 13 CNMR(101MHz,DMSO)δ165.45,152.20,147.42,136.63,135.15,133.06,129.83,129.42 ,123.88,123.77,118.48,115.66,113.32,106.31,44.24,44.04,37.67,25.45,22.29.

[0283] Example 16

[0284]

[0285] Step 1: {[4-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)-2-nitrophenyl]amino}acetic acid-2-methylpropan-2-yl ester (16a)

[0286] The reactant S1 (200 mg, 0.78 mmol) was dissolved in 4 mL DMF, and glycine tert-butyl ester (154 mg, 1.17 mmol) and triethylamine (239 mg, 2.34 mmol) were added. The reaction solution was heated at 90 ° C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. After flash purification (PE / EA = 10:1), the product 16a (123 mg, yield 43%) was obtained.

[0287] LC-MS: m / z=368.2[M+H] +

[0288] 1 H NMR (400MHz, CDCl3) δ8.24(t,J=5.2Hz,1H),8.11(d,J=2.7Hz,1H),7.63(s,1H),6 .65(d,J=9.2Hz,1H),6.54(s,1H),3.98(d,J=5.2Hz,2H),1.52(d,J=1.1Hz,18H). 13 C NMR (101MHz, CDCl3) δ168.30,153.13,140.87,131.84,129.22,127.72,114.25,82.85,45.75,28.32,28.04.

[0289] Step 2: [2-(3,5-dichloro-2-hydroxyphenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]acetate-2-methylpropan-2-yl ester (16b)

[0290] The reactant 16a (123 mg, 0.34 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (76 mg, 0.40 mmol), sodium dithionite (175 mg, 1.01 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=3:1) to obtain the product 16b (76 mg, yield 45%).

[0291] LC-MS: m / z=508.1[M+H] +

[0292] 1 H NMR (400MHz, CDCl3) δ7.81(d,J=1.9Hz,1H),7.49–7.44(m,2H),7.41(d,J=8.4Hz,1H),7.28(s,1H),4.95(s,2H),1.57(s,9H),1.53(s,9H). 13 CNMR(101MHz,CDCl3)δ166.11,153.90,150.27,140.14,135.01,131.80,131.42 ,124.33,123.98,123.18,117.29,114.54,109.68,84.37,48.33,28.39,27.94.

[0293] Step 3: {5-[(2-chloroacetyl)amino]-2-(3,5-dichloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl}acetate-2-methylpropan-2-yl ester (XD16)

[0294] The reactant 16b (76 mg, 0.15 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring, and the reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (61 mg, 0.60 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (20 mg, 0.18 mmol) was added dropwise while stirring under ice bath, and the reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD16 (38 mg, yield 59%).

[0295] LC-MS: m / z=427.8[M+H] +

[0296] 1 H NMR (400MHz, DMSO) δ10.45(s,1H),7.77(d,J=2.5Hz,1H),7.71(d,J=8.8Hz,1H),7.59–7.47(m,2H),5.23(s,2H),4.31(s,2H),2.08(s,1H). 13CNMR(101MHz,DMSO)δ169.61,165.05,152.26,150.39,140.35,134.82,132. 66,131.59,127.87,123.43,118.29,117.38,111.66,109.44,46.93,44.11.

[0297] Embodiment 17

[0298]

[0299] Step 1: {[4-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)-2-nitrophenyl]amino}acetic acid-2-methylpropan-2-yl ester (17a)

[0300] The reactant S1 (300 mg, 1.17 mmol) was dissolved in 4 mL DMF, and leucine methyl ester hydrochloride (319 mg, 1.76 mmol) and triethylamine (474 ​​mg, 4.69 mmol) were added. The reaction solution was heated at 90 ° C for overnight reaction. The reaction solution was then extracted three times with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. After flash purification (PE / EA = 10:1), the product 17a (148 mg, yield 33%) was obtained.

[0301] LC-MS: m / z=382.1[M+H] +

[0302] 1 H NMR (400MHz, CDCl3) δ8.11(d,J=2.6Hz,1H),8.02(d,J=7.5Hz,1H),7.59(s,1H),6.71(d,J=9.2Hz,1H),6.57(s,1H), 4.22(q,J=7.0Hz,1H),3.75(s,3H),1.82(dt,J=7.2,2.3Hz,3H),1.52(s,9H),1.04–1.00(m,3H),0.96–0.93(m,3H). 13 C NMR (101MHz, CDCl3) δ173.42,153.11,140.87,132.04,129.32,127.96,114.20,54.68,52.50,41.66,28.31,24.96,22.75,21.93.

[0303] Step 2: 2-[2-(3,5-dichloro-2-hydroxyphenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid methyl ester (17b)

[0304] The reactant 17a (148 mg, 0.39 mmol), 3,5-dichloro-2-hydroxybenzene-1-carboxaldehyde (88 mg, 0.47 mmol), sodium dithionite (203 mg, 1.16 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=5:1) to obtain the product 17b (130 mg, yield 64%).

[0305] LC-MS: m / z=521.8[M+H] +

[0306] 1 H NMR (400MHz, CDCl3) δ7.67(d,J=2.0Hz,1H),7.46(t,J=1.9Hz,2H),7.44(s,1H),7.29(t,J=4.1Hz,1H),6.66(s,1H),5.27(dd,J=9.8 ,5.6Hz,1H),3.87(s,3H),2.28–2.00(m,2H),1.57(s,9H),1.28(td,J=6.1,3.4Hz,1H),0.70(d,J=6.7Hz,3H),0.61(d,J=6.5Hz,3H). 13 C NMR (101MHz, CDCl3) δ169.96,153.16,152.70,150.69,141.51,134.54,131.69,129.47,126.01,124. 45,124.03,117.05,116.00,112.60,57.87,53.19,38.61,28.39,26.92,24.48,22.66,22.54,21.11.

[0307] Step 3: 2-{5-[(2-chloroacetyl)amino]-2-(3,5-dichloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl}-4-methylpentanoate (XD17)

[0308] The reactant 17b (130 mg, 0.25 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (101 mg, 1.0 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (34 mg, 0.30 mmol) was added dropwise while stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was complete as monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD17 (66 mg, yield 53%).

[0309] LC-MS: m / z=498.0[M+H] +

[0310] 1 H NMR (400MHz, DMSO) δ10.60(s,1H),8.21(d,J=1.9Hz,1H),7.86(d,J=2.6Hz,1H),7.73(d,J=8.9Hz,1H),7.60(d,J=2.6Hz,1H),7.53(dd,J=8.9,1.9H z,1H),5.01(dd,J=10.3,5.1Hz,1H),4.33(s,2H),3.74(s,3H),2.30–1.9 3(m,2H),1.10–0.98(m,1H),0.66(d,J=6.6Hz,3H),0.57(d,J=6.5Hz,3H). 13 CNMR(101MHz,DMSO)δ169.73,165.27,151.55,149.40,139.21,135.40,132.71,130.39,129.68,1 24.17,123.56,118.48,117.75,114.42,108.46,57.97,53.29,44.06,38.53,24.52,22.77,21.14.

[0311] Embodiment 18

[0312]

[0313] The reaction steps are the same as those in Example 9. The fluorine atom of the initial reactant is replaced by an amine reagent after the SNAr reaction. The intermediate containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite. The intermediate is then deprotected from Boc and reacted with chloroacetyl chloride. The target product is finally purified by HPLC to obtain the target product.

[0314] 1H NMR (500MHz, DMSO) δ10.68(s,1H),8.26(d,J=2.0Hz,1H),7.89(d,J=2.6Hz,1H),7.79(d,J=9.0Hz,1H),7.64(d,J=2.6Hz,1H),7.56(dd,J=9. 1,2.0Hz,1H),4.96(dd,J=10.2,5.2Hz,1H),4.33(s,2H),2.29–1.96(m,2H),1.16–1.04(m,1H),0.68(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H). 13 C NMR (126MHz, DMSO) δ170.41,165.40,151.81,148.70,137.26,135.96,133.14,130.29,129.13 ,124.14,123.87,118.13,117.25,115.08,107.35,58.45,44.04,38.57,24.62,22.78,21.21.

[0315] Embodiment 19

[0316]

[0317] The reaction steps are the same as those in Example 9. After the initial reactants are subjected to SNAr reaction, the fluorine atom is replaced by an amine reagent, and then the intermediate containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite. After de-Boc protection, the target product is purified by HPLC.

[0318] 1 H NMR (500MHz, DMSO) δ10.69(s,1H),8.26(d,J=2.0Hz,1H),7.89(d,J=2.6Hz,1H),7.79(d,J=9.0Hz,1H),7.63(d,J=2.6Hz,1H),7.56(dd,J=9. 0,1.9Hz,1H),4.96(dd,J=10.1,5.2Hz,1H),4.34(s,2H),2.26–1.99(m,2H),1.16–1.04(m,1H),0.68(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H). 13C NMR (126MHz, DMSO) δ170.43,165.40,151.81,148.73,137.36,135.93,133.11,130.28,129.16 ,124.14,123.87,118.11,117.32,115.05,107.41,58.43,44.04,38.57,24.62,22.78,21.21.

[0319] Embodiment 20

[0320]

[0321] Step 1: 2-[2-(3-chloro-2-hydroxyphenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (20a)

[0322] The reactant S2 (150 mg, 0.35 mmol), 3-chloro-2-hydroxybenzene-1-carboxaldehyde (82 mg, 0.53 mmol), sodium dithionite (185 mg, 1.06 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA = 10:1) to obtain the product 20a (71 mg, yield 38%).

[0323] LC-MS: m / z=529.8[M+H] +

[0324] 1 H NMR (400MHz, CDCl3) δ7.78(d,J=2.0Hz,1H),7.58(dd,J=7.9,1.5Hz,1H),7.50–7.46(m,2H),7.36–7.29(m,1H),6.94(t,J=7.9Hz,1H),6.68(s, 1H),5.32(dd,J=9.9,5.5Hz,1H),1.56(s,9H),1.47(s,9H),1.30–1.26( m,1H),0.91–0.85(m,2H),0.67(d,J=6.7Hz,3H),0.56(d,J=6.6Hz,3H). 13C NMR (101MHz, CDCl3) δ168.80,154.15,153.14,152.14,141.81,134.46,132.07,129.72,126.04,123. 13,119.31,116.37,114.79,112.84,83.42,58.97,38.43,28.39,27.96,27.92,24.54,22.65,21.03.

[0325] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-(3-chloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD20)

[0326] The reactant 20a (71 mg, 0.13 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring, and the reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (54 mg, 0.54 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (18 mg, 0.16 mmol) was added dropwise while stirring under ice bath, and the reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD20 (48 mg, yield 80%).

[0327] LC-MS: m / z=450.1[M+H] +

[0328] 1 H NMR (500MHz, DMSO) δ10.69(s,1H),8.26(d,J=2.1Hz,1H),7.80(d,J=9.0Hz,1H),7.74(dd,J=8.0,1.6Hz,1H),7.55(td,J=8.5,1.8Hz,2H),7.14( t,J=7.9Hz,1H),4.98(dd,J=10.4,5.0Hz,1H),4.34(s,2H),2.28–1.97(m,2H),1.10–0.99(m,1H),0.65(d,J=6.6Hz,3H),0.53(d,J=6.5Hz,3H). 13 C NMR (126MHz, DMSO) δ170.43,165.43,152.47,149.91,136.03,134.10,130.71,128.92,12 2.63,121.52,118.06,115.70,115.15,107.04,58.44,44.05,38.54,24.63,22.79,21.07.

[0329] Embodiment 21

[0330]

[0331] Step 1: 2-[2-(5-chloro-2-hydroxyphenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (21a)

[0332] The reactant S2 (150 mg, 0.35 mmol), 5-chloro-2-hydroxybenzene-1-carboxaldehyde (82 mg, 0.53 mmol), sodium dithionite (185 mg, 1.06 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. The reaction was monitored by LC-MS to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA=20:1) to obtain the product 21a (45 mg, yield 24%).

[0333] LC-MS: m / z=430.1[M+H] +

[0334] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-(5-chloro-2-hydroxyphenyl)benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD21)

[0335] The reactant 21a (45 mg, 0.10 mmol) was dissolved in 2 mL of dichloromethane, and 1 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring, and the reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (42 mg, 0.42 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (14 mg, 0.13 mmol) was added dropwise while stirring under ice bath, and the reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD21 (34 mg, yield 72%).

[0336] LC-MS: m / z=450.1[M+H] +

[0337] 1H NMR(500MHz,DMSO)δ10.71(s,1H),8.24(d,J=2.0Hz,1H),7.81(d,J=9.0Hz,1H),7 .64(d,J=2.7Hz,1H),7.57(ddd,J=12.7,8.9,2.3Hz,2H),7.15(d,J=8.9Hz,1H),4 .93(dd,J=10.8,4.8Hz,1H),4.33(s,2H),2.30(ddd,J=14.8,10.9,4.3Hz,1H),2. 01–1.91(m,1H),1.08–0.96(m,1H),0.67(d,J=6.6Hz,3H),0.54(d,J=6.5Hz,3H). 13 C NMR (126MHz, DMSO) δ170.60,165.44,155.65,149.52,135.96,133.84,131.57,123.47 ,121.46,118.67,117.98,115.52,106.87,58.66,44.03,38.56,24.64,22.86,20.90.

[0338] Embodiment 22

[0339]

[0340] Step 1: 2-[2-(3,5-dichlorophenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (22a)

[0341] The reactant S2 (150 mg, 0.35 mmol), 3,5-dichlorobenzene-1-carboxaldehyde (93 mg, 0.53 mmol), sodium dithionite (185 mg, 1.06 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA = 10:1) to obtain the product 22a (105 mg, yield 54%).

[0342] LC-MS: m / z=548.2[M+H] +

[0343] 1H NMR(500MHz, CDCl3)δ7.71–7.67(m,3H),7.55–7.45(m,3H),6.67(s,1H),4.96(dd,J=10.3,5.2Hz,1H),1.5 5(s,9H),1.46(s,9H),1.34–1.30(m,1H),0.90–0.85(m,2H),0.71(d,J=6.6Hz,3H),0.60(d,J=6.6Hz,3H). 13 C NMR (126MHz, CDCl3) δ171.18,168.55,153.19,152.18,143.61,135.57,134.17,133.15,130. 38,130.00,127.99,116.65,112.35,83.48,58.20,38.00,28.38,27.90,24.40,22.78,21.21.

[0344] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-(3,5-dichlorophenyl)benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD22)

[0345] The reactant 22a (105 mg, 0.19 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (78 mg, 0.77 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (26 mg, 0.23 mmol) was added dropwise while stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was complete as monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD22 (59 mg, yield 66%).

[0346] LC-MS: m / z=468.1[M+H] +

[0347] 1 H NMR (500MHz, DMSO) δ10.51(s,1H),8.16(d,J=2.0Hz,1H),7.91(t,J=1.9Hz,1H),7.77(d,J=2.0Hz,2H),7.64(d,J=8.9Hz,1H),7.51(dd,J=8. 9,2.0Hz,1H),5.24(dd,J=10.2,5.3Hz,1H),4.31(s,2H),2.17–1.96(m,2H),1.05–0.93(m,1H),0.65(d,J=6.7Hz,3H),0.57(d,J=6.5Hz,3H).13 C NMR (126MHz, DMSO) δ171.16,165.14,151.29,141.15,135.24,135.00,132.40,13 0.72,128.56,117.61,113.23,109.78,57.57,44.07,37.96,24.58,22.88,21.39.

[0348] Embodiment 23

[0349]

[0350] Step 1: 2-[2-(3-chlorophenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (23a)

[0351] The reactant S2 (150 mg, 0.35 mmol), 3-chlorobenzene-1-carboxaldehyde (74 mg, 0.53 mmol), sodium dithionite (185 mg, 1.06 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. The reaction was monitored by LC-MS. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA = 10:1) to obtain the product 23a (120 mg, yield 66%).

[0352] LC-MS: 514.2 [M+H] +

[0353] 1 H NMR(500MHz, CDCl3)δ7.78(t,J=1.8Hz,1H),7.65(dd,J=7.2,1.7Hz,2H),7.53–7.43(m,4H),6.61(s,1H),5.01(dd,J=10.4,5 .1Hz,1H),1.55(s,9H),1.46(s,9H),1.33(d,J=2.7Hz,1H),1.02–0.87(m,2H),0.69(d,J=6.7Hz,3H),0.55(d,J=6.6Hz,3H). 13C NMR (126MHz, CDCl3) δ168.83,153.61,143.69,134.75,133.92,132.05,130.35,130.16,1 30.09,129.70,127.78,112.23,83.25,58.10,38.12,28.40,27.92,24.38,22.82,21.13.

[0354] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-(3-chlorophenyl)benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD23)

[0355] The reactant 23a (120 mg, 0.23 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (95 mg, 0.94 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (32 mg, 0.28 mmol) was added dropwise while stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was complete as monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD23 (51 mg, yield 50%).

[0356] LC-MS: m / z=434.1[M+H] +

[0357] 1 H NMR (500MHz, DMSO) δ10.62(s,1H),8.22(d,J=2.0Hz,1H),7.84(t,J=1.9Hz,1H),7.79–7.66(m,4H),7.55(dd,J=9.0,2.0Hz,1H) ,5.25(dd,J=10.3,5.2Hz,1H),4.32(s,2H),2.23–1.93(m,2H),1.06–0.94(m,1H),0.64(d,J=6.7Hz,3H),0.53(d,J=6.5Hz,3H). 13 C NMR (126MHz, DMSO) δ170.89,165.29,151.86,138.86,135.63,134.29,131.69,131.62,12 9.82,129.50,128.71,117.91,113.79,108.48,57.84,44.05,38.00,24.55,22.86,21.21.

[0358] Embodiment 24

[0359]

[0360] Step 1: 2-[2-(3,5-dichloro-2-methoxyphenyl)-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl]-4-methylpentanoic acid-2-methylpropan-2-yl ester (24a)

[0361] The reactant S2 (150 mg, 0.35 mmol), 3,5-dichloro-2-methoxybenzene-1-carboxaldehyde (109 mg, 0.53 mmol), sodium dithionite (185 mg, 1.06 mmol) were dissolved in 4 mL DMSO, and the reaction solution was stirred at 100 ° C overnight. LC-MS monitored the reaction to be complete. The reaction solution was cooled to room temperature and ice water was added. A large amount of solid precipitated. The solid was filtered and washed with cold water three times. The crude product was flash purified (PE / EA = 10:1) to obtain the product 24a (138 mg, yield 67%).

[0362] LC-MS: 577.8 [M+H] +

[0363] 1 H NMR(400MHz, CDCl3)δ7.73(t,J=1.3Hz,1H),7.56(d,J=2.3Hz,2H),7.49–7.37(m,2H),6.66(s,1H) ,4.65(s,1H),3.45(s,3H),1.55(s,18H),1.29–1.24(m,1H),1.07–0.77(m,2H),0.77–0.47(m,6H). 13 C NMR (126MHz, DMSO) δ171.02,165.13,153.50,149.07,141.54,134.62,132.88,131.20,13 0.51,129.42,129.35,117.36,109.86,62.32,57.89,44.06,38.53,24.70,22.84,21.08.

[0364] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-(3,5-dichloro-2-methoxyphenyl)benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD24)

[0365] The reactant 24a (138 mg, 0.24 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (97 mg, 0.96 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (32 mg, 0.29 mmol) was added dropwise while stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was complete as monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD24 (63 mg, yield 53%).

[0366] LC-MS: m / z=498.1[M+H] +

[0367] 1 H NMR (500MHz, DMSO) δ10.49(s,1H),8.15(d,J=1.9Hz,1H),7.97(d,J=2.6Hz,1H),7.64(d,J=8.9Hz,2H),7.47(dd,J=8.9,2.0Hz,1H),4. 81(dd,J=10.6,5.0Hz,1H),4.31(s,2H),3.70–3.32(m,3H),2.24–1.93(m,2H),0.98(s,1H),0.64(d,J=6.6Hz,3H),0.59–0.47(m,3H). 13 C NMR (126MHz, DMSO) δ171.02,165.13,153.50,149.07,141.54,134.62,132.88,131.20,13 0.51,129.42,129.35,117.36,109.86,62.32,57.89,44.06,38.53,24.70,22.84,21.08.

[0368] Embodiment 25

[0369]

[0370] The reaction steps are the same as those in Example 6. After the initial reactants are subjected to SNAr reaction, the fluorine atoms are replaced by amine reagents, and then the intermediate containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0371] 1H NMR (500MHz, MeOD) δ8.49(d,J=2.6Hz,1H),8.35(d,J=2.0Hz,1H),8.06(d,J=2.7Hz,1H),7.81(d,J=9.0Hz,1H),7.60(d d,J=9.0,2.0Hz,1H),4.28(s,2H),2.41–2.13(m,2H),1.27–1.15(m,1H),0.79(d,J=6.6Hz,3H),0.72(d,J=6.5Hz,3H). 13 C NMR(126MHz,MeOD)δ170.25,166.33,151.29,147.43,138.19,137.16,136.11,135.68,129.25 ,128.46,124.40,119.09,118.38,114.47,107.62,58.68,42.65,38.46,24.57,21.58,20.11.

[0372] Embodiment 26

[0373]

[0374] The reaction steps are the same as those in Example 6. After the initial reactants are subjected to SNAr reaction, the fluorine atoms are replaced by amine reagents, and then the intermediate containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0375] 1 H NMR(500MHz,MeOD)δ8.39(d,J=1.7Hz,1H),7.84(d,J=9.0Hz,1H),7.69–7.57(m,2H),7.51(s,1H),5.14(dd,J=10.8,4.6Hz,1H),4.28(s,2H),2. 41(ddd,J=14.8,10.9,4.2Hz,1H), 2.13(ddd,J=14.4,9.7,4.7Hz,1H), 1.13(dt,J=6.8,3.8Hz,1H), 0.78(d,J=6.7Hz,3H), 0.67(d,J=6.5Hz,3H). 13C NMR(126MHz,MeOD)δ170.34,166.38,152.67,150.71,148.18,144.00,136.09,135.60,128.77,126.34,124.22,1 24.15,120.30,120.12,118.45,114.94,106.69,58.97,47.45,47.28,47.11,42.66,38.46,24.53,21.60,19.74.

[0376] Embodiment 27

[0377]

[0378] The reaction steps are the same as those in Example 6. After the initial reactants are subjected to SNAr reaction, the fluorine atoms are replaced by amine reagents, and then the intermediate containing a benzimidazole skeleton is formed by a one-pot ring closure under the action of hydrosulfite. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0379] 1 H NMR (500MHz, MeOD) δ8.35 (s, 1H), 7.97–7.87 (m, 1H), 7.80 (d, J = 9.0Hz, 1H), 7. 73(dd,J=15.8,2.3Hz,1H),7.63–7.51(m,1H),5.08(dd,J=10.6,4.8Hz,2H),4. 27(d,J=1.7Hz,2H),2.43–2.33(m,1H),2.13(ddd,J=14.5,9.6,4.9Hz,1H),1.1 5(dtd,J=13.0,10.9,6.4Hz,1H),0.78(d,J=6.6Hz,3H),0.68(d,J=6.4Hz,3H).

[0380] Embodiment 28

[0381]

[0382] Step 1: 2-(2-{3-chloro-2-hydroxy-5-[2-(trifluoromethyl)phenyl]phenyl}-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl)-4-methylpentanoic acid-2-methylpropan-2-yl ester (28a)

[0383] The reactants N-[2-amino-4-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)phenyl]-DL-leucine-2-methylpropan-2-yl ester (110 mg, 0.28 mmol), 3-chloro-2-hydroxy-5-[2-(trifluoromethyl)phenyl]benzene-1-carboxaldehyde (92 mg, 0.31 mmol), sodium metabisulfite (161 mg, 0.84 mmol) were dissolved in 4 mL of DMF, and the reaction solution was stirred at 100°C for 3 h. The reaction was monitored to be complete by LC-MS. The reaction solution was cooled to room temperature, and ice water was added. A large amount of solid precipitated. The solid was filtered and rinsed with cold water three times. The crude product was flash purified (PE / EA=5:1) to obtain product 28a (177 mg, yield 94%).

[0384] LC-MS: 673.8[M+H] +

[0385] 1 H NMR(400MHz, CDCl3)δ7.84(d,J=2.0Hz,1H),7.82–7.77(m,1H),7.66–7.56(m, 2H),7.55–7.49(m,2H),7.46(d,J=2.0Hz,1H),7.40–7.29(m,2H),6.64(s,1H) ,5.38(dt,J=12.0,5.9Hz,1H),2.26–1.85(m,2H),1.57(s,9H),1.26(s,9H),0 .81(pd,J=6.4,4.4Hz,1H), 0.67(dd,J=9.1,6.6Hz,3H), 0.56(d,J=6.5Hz,3H). 13 C NMR (101MHz, CDCl3) δ168.75,153.80,153.12,151.90,141.87,139.34,134.53,132.38,132.22,131.61,130.97,129.75,127.94, 126.67,126.02,122.52,116.53,113.75,113.09,109.39,83.16,80.63,77.24,58.84,38.31,28.38,27.58,24.38,22.70,20.61.

[0386] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-{3-chloro-2-hydroxy-5-[2-(trifluoromethyl)phenyl]phenyl}benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD28)

[0387] The reactant 28a (177 mg, 0.26 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (106 mg, 1.1 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (35 mg, 0.32 mmol) was added dropwise under stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD28 (94 mg, yield 60%).

[0388] LC-MS: m / z=594.1[M+H] +

[0389] 1 H NMR (400MHz, DMSO) δ10.65(s,1H),8.23(d,J=2.0Hz,1H),7.85(dd,J=8.0,1. 3Hz,1H),7.75(dd,J=8.2,6.1Hz,2H),7.68–7.61(m,2H),7.56(dd,J=9.0,2.0 Hz,1H),7.51–7.43(m,2H),5.04(dd,J=10.5,4.8Hz,1H),4.33(s,2H),2.31– 1.95(m,2H),1.17–0.98(m,1H),0.65(d,J=6.6Hz,3H),0.57(d,J=6.5Hz,3H). 13 C NMR (101MHz, DMSO) δ170.66,165.33,152.30,149.84,138.48,138.21,135.65,133.68,132.94,132.80,132.16,130.62,129.38,12 9.06,127.65,127.36,126.64,125.93,123.20,122.24,117.82,116.00,114.71,107.87,58.26,44.05,38.58,24.62,22.80,20.93.

[0390] Embodiment 29

[0391]

[0392] Step 1: 2-(2-{3-chloro-2-hydroxy-5-[3-(trifluoromethyl)phenyl]phenyl}-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl)-4-methylpentanoic acid-2-methylpropan-2-yl ester (29a)

[0393] The reactants N-[2-amino-4-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)phenyl]-DL-leucine-2-methylpropan-2-yl ester (110 mg, 0.28 mmol), 3-chloro-2-hydroxy-5-[3-(trifluoromethyl)phenyl]benzene-1-carboxaldehyde (92 mg, 0.31 mmol), sodium metabisulfite (161 mg, 0.84 mmol) were dissolved in 4 mL of DMF, and the reaction solution was stirred at 100°C for 3 h. The reaction was monitored to be complete by LC-MS. The reaction solution was cooled to room temperature, and ice water was added. A large amount of solid precipitated. The solid was filtered and rinsed with cold water three times. The crude product was flash purified (PE / EA=5:1) to obtain product 29a (137 mg, yield 73%).

[0394] LC-MS: 673.8[M+H] +

[0395] 1 H NMR (400MHz, CDCl3) δ7.85–7.80(m,3H),7.80–7.76(m,1H),7.74(d,J=2.2Hz,1H),7.67 –7.56(m,2H),7.52(d,J=8.8Hz,1H),7.34(d,J=8.8Hz,1H),6.68(s,1H),5.34(dd,J=9.9 ,5.4Hz,1H),2.24(ddd,J=14.5,9.9,4.9Hz,1H),1.96(ddd,J=14.4,9.1,5.4Hz,1H),1. 57(s,9H),1.44(s,9H),1.30–1.26(m,1H),0.67(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H). 13 C NMR (101MHz, CDCl3) δ168.74,154.10,153.13,151.80,141.77,140.01,134.60,131.62,131.30,130.63,129.99,129.75,129.50,125.41, 124.70,124.18,123.90,123.39,123.35,116.62,115.27,112.93,10 9.43,83.64,80.64,59.17,38.34,28.39,27.84,24.59,22.61,21.04.

[0396] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-{3-chloro-2-hydroxy-5-[3-(trifluoromethyl)phenyl]phenyl}benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD29)

[0397] The reactant 29a (137 mg, 0.20 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring, and the reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (82 mg, 0.81 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (27 mg, 0.24 mmol) was added dropwise while stirring under ice bath, and the reaction was allowed to react at 0°C for 4 h. The reaction was monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD29 (69 mg, yield 58%).

[0398] LC-MS: m / z=594.1[M+H] +

[0399] 1 H NMR (400MHz, DMSO) δ10.71(s,1H),8.29(d,J=2.0Hz,1H),8.21(d,J=2.3Hz,1H),8.10–7.97(m,3H),7.83(d,J=9.0Hz,1H),7.77–7.66(m,2H),7.59(d d,J=9.0,2.0Hz,1H),5.06(dd,J=10.3,5.1Hz,1H),4.35(s,2H),2.33–1.9 7(m,2H),1.19–1.05(m,1H),0.67(d,J=6.6Hz,3H),0.56(d,J=6.5Hz,3H). 13 C NMR (101MHz, DMSO) δ170.54,165.42,152.59,149.77,139.04,136.01,132.13,131.60,130.86,130.60,130.26 ,129.23,129.09,125.97,124.76,123.45,118.08,115.10,107.20,58.55,44.05,38.54,24.59,22.84,21.14.

[0400] Embodiment 30

[0401]

[0402] Step 1: 2-(2-{3-chloro-2-hydroxy-5-[4-(trifluoromethyl)phenyl]phenyl}-5-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)benzo[d]imidazol-1-yl)-4-methylpentanoic acid-2-methylpropan-2-yl ester (30a)

[0403] The reactants N-[2-amino-4-({[(2-methylpropan-2-yl)oxy]carbonyl}amino)phenyl]-DL-leucine-2-methylpropan-2-yl ester (110 mg, 0.28 mmol), 3-chloro-2-hydroxy-5-[4-(trifluoromethyl)phenyl]benzene-1-carboxaldehyde (92 mg, 0.31 mmol), sodium metabisulfite (161 mg, 0.84 mmol) were dissolved in 4 mL of DMF, and the reaction solution was stirred at 100°C for 3 h. The reaction was monitored to be complete by LC-MS. The reaction solution was cooled to room temperature, and ice water was added. A large amount of solid precipitated. The solid was filtered and rinsed with cold water three times. The crude product was flash purified (PE / EA=5:1) to obtain product 30a (156 mg, yield 83%).

[0404] LC-MS: 673.8[M+H] +

[0405] 1 H NMR (400MHz, CDCl3) δ7.82 (dd, J=8.9, 2.1Hz, 2H), 7.75 (d, J=2.2Hz, 1H), 7.74–7.69 (m ,4H),7.54(d,J=8.8Hz,1H),7.35(d,J=8.8Hz,1H),6.67(s,1H),5.34(dd,J=10.1,5.3 Hz,1H),2.27(ddd,J=14.6,10.1,4.8Hz,1H),1.94(ddd,J=14.4,9.1,5.3Hz,1H),1.58 (s,9H),1.47(s,9H),1.32–1.27(m,1H),0.66(d,J=6.6Hz,3H),0.57(d,J=6.6Hz,3H). 13 C NMR (101MHz, CDCl3) δ168.85,154.26,153.13,151.81,142.60,141.77,134.61,131.12,130.61,129.79,129.43,126.88,125.94,1 25.90,125.52,124.81,123.89,122.82,116.63,115.18,113.04,109.43,83.64,59.27,38.40,28.39,27.95,24.55,22.63,21.01.

[0406] Step 2: 2-{5-[(2-chloroacetyl)amino]-2-{3-chloro-2-hydroxy-5-[4-(trifluoromethyl)phenyl]phenyl}benzo[d]imidazol-1-yl}-4-methylpentanoic acid (XD30)

[0407] The reactant 30a (156 mg, 0.23 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added dropwise to the reaction solution under stirring. The reaction was allowed to react at room temperature for 4 h. After the reaction was complete as detected by LCMS, the solvent was dried under reduced pressure. 4 mL of dichloromethane and triethylamine (94 mg, 0.93 mmol) were added to the reaction bottle after drying, and chloroacetyl chloride (29 mg, 0.25 mmol) was added dropwise while stirring under ice bath. The reaction was allowed to react at 0°C for 4 h. The reaction was complete as monitored by TLC spot plate. The crude product was purified by Pre-HPLC to obtain the final product XD30 (73 mg, yield 53%).

[0408] LC-MS: m / z=594.1[M+H] +

[0409] 1 H NMR (400MHz, DMSO) δ10.78(s,1H),8.34(d,J=2.1Hz,1H),8.18(d,J=2.3Hz,1H),8.04–7.92(m,3H),7.88(d,J=9.0Hz,1H),7.81(d,J=8.3Hz,2H),7.63 (dd,J=9.0,2.0Hz,1H),5.08(dd,J=10.4,5.0Hz,1H),4.36(s,2H),2.34–1. 98(m,2H),1.18–1.04(m,1H),0.67(d,J=6.6Hz,3H),0.56(d,J=6.5Hz,3H). 13 C NMR (101MHz, DMSO) δ170.41,165.50,152.82,149.41,141.89,136.38,136.06,132.41,131.67,129.44,128.83,128.43,12 7.56,126.33,126.29,126.07,123.57,123.36,118.36,115.80,115.36,106.70,58.72,44.03,38.53,24.59,22.80,21.07.

[0410] Embodiment 31

[0411]

[0412] The reaction steps are the same as those in Example 28. The initial reactants are cyclized under the action of sodium pyrosulfite to form an intermediate containing a benzimidazole skeleton. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0413] 1 H NMR(500MHz,MeOD)δ8.35(s,1H),7.93(s,1H),7.78(d,J=7.4Hz,2H),7.58(d,J=8.4H z,1H),7.33(d,J=7.4Hz,1H),7.20(t,J=7.5Hz,1H),6.93(t,J=6.8Hz,2H),5.28(dd,J =10.8,3.8Hz,1H),4.28(s,2H),3.38(s,1H),2.46–2.36(m,1H),2.17–2.02(m,1H),1 .42–1.27(m,3H),1.12(d,J=5.6Hz,1H),0.76(d,J=6.5Hz,3H),0.66(d,J=6.3Hz,3H).

[0414] Embodiment 32

[0415]

[0416] The reaction steps are the same as those in Example 28. The initial reactants are cyclized under the action of sodium pyrosulfite to form an intermediate containing a benzimidazole skeleton. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0417] 1 H NMR(500MHz,MeOD)δ8.42(s,1H),7.98(d,J=1.9Hz,1H),7.87(d,J=9.0Hz,1H),7.80(d,J=1 .9Hz,1H),7.65–7.52(m,1H),7.30(t,J=7.9Hz,1H),7.11(d,J=7.7Hz,1H),7.05(s,1H),6.8 3(dd,J=8.0,2.1Hz,1H),5.20(dd,J=10.9,4.4Hz,2H),4.28(s,2H),2.42(s,1H),2.20–1.97 (m,2H),1.40–1.26(m,1H),1.24–1.10(m,2H),0.78(d,J=6.6Hz,3H),0.67(d,J=6.5Hz,3H).

[0418] Embodiment 33

[0419]

[0420] The reaction steps are the same as those in Example 28. The initial reactants are cyclized under the action of sodium pyrosulfite to form an intermediate containing a benzimidazole skeleton. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0421] 1 H NMR(500MHz,MeOD)δ8.44(s,1H),8.30(s,1H),8.15–8.03(m,2H),7.97–7.84( m,3H),7.70–7.55(m,2H),5.23(dd,J=10.8,4.6Hz,1H),4.29(s,2H),2.42(ddd ,J=14.7,10.9,4.2Hz,1H),2.16(ddd,J=14.5,9.7,4.7Hz,1H),1.46–1.28(m, 4H), 1.20 (dt, J=6.8, 3.7Hz, 1H), 0.78 (d, J=6.6Hz, 3H), 0.68 (d, J=6.5Hz, 3H).

[0422] Embodiment 34

[0423]

[0424] The reaction steps are the same as those in Example 28. The initial reactants are cyclized under the action of sodium pyrosulfite to form an intermediate containing a benzimidazole skeleton. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0425] 1 H NMR(500MHz,MeOD)δ8.40(s,1H),8.00(d,J=1.9Hz,1H),7.92–7.80(m,2H),7.61(d,J=9.0Hz,1H ),7.53(s,1H),7.51–7.40(m,2H),7.29(d,J=7.2Hz,1H),5.30–5.09(m,2H),4.28(s,2H),3.36– 3.29(m,4H),3.03(s,3H),2.41(ddd,J=14.8,10.9,4.2Hz,1H),2.22(t,J=7.6Hz,1H),2.17–2.0 0(m,2H),1.20–1.12(m,1H),0.92(t,J=6.6Hz,1H),0.77(d,J=6.6Hz,3H),0.66(d,J=6.5Hz,3H).

[0426] Embodiment 35

[0427]

[0428] The reaction steps are the same as those in Example 28. The initial reactants are cyclized under the action of sodium pyrosulfite to form an intermediate containing a benzimidazole skeleton. After de-Boc protection, the intermediate is reacted with chloroacetyl chloride and then purified by HPLC to obtain the target product.

[0429] 1 H NMR(500MHz,MeOD)δ8.40(s,1H),7.94(s,1H),7.84(d,J=9.0Hz,1H),7.81–7.74(m,1H),7.60(d,J=8.9 Hz,1H),7.18(s,1H),7.09(d,J=8.1Hz,1H),6.89(dd,J=8.2,1.7Hz,1H),5.20(dd,J=10.9,4.5Hz,1H),4 .28(s,2H),3.94(d,J=3.9Hz,3H),2.41(ddd,J=14.7,11.2,4.0Hz,1H),2.17–2.00(m,2H),1.42–1.30( m,3H),1.14(dt,J=6.7,3.7Hz,1H),0.93(t,J=6.8Hz,1H),0.77(d,J=6.6Hz,3H),0.66(d,J=6.4Hz,3H).

[0430] Embodiment 36

[0431]

[0432] The reactant S3 (50 mg, 0.12 mmol) was dissolved in 4 mL of dichloromethane, stirred under ice bath and acryloyl chloride (12 mg, 0.14 mmol) was added dropwise, reacted at 0°C for 4 h, and the reaction was complete after TLC spot plate monitoring. The crude product was purified by Pre-HPLC to obtain the final product XD36 (29 mg, yield 51%).

[0433] LC-MS: m / z=462.1[M+H] +

[0434] 1H NMR (400MHz, DMSO) δ10.53(s,1H),8.38(d,J=2.0Hz,1H),7.88(d,J=2.6Hz,1H),7. 77(d,J=9.0Hz,1H),7.65–7.57(m,2H),6.50(dd,J=16.9,10.1Hz,1H),6.32(dd,J= 17.0,2.0Hz,1H),5.80(dd,J=10.0,2.0Hz,1H),4.95(dd,J=10.2,5.2Hz,1H),2.27 –1.98(m,2H),1.09(q,J=6.9Hz,1H),0.68(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H). 13 C NMR (101MHz, DMSO) δ170.43,163.92,151.83,148.58,137.34,136.48,133.09,132.15,130.24,128 .94,127.67,124.13,123.84,118.14,117.28,114.93,107.24,58.42,38.56,24.62,22.77,21.19.

[0435] Embodiment 37

[0436]

[0437] The reactant S3 (50 mg, 0.12 mmol) was dissolved in 4 mL of dichloromethane, stirred under ice bath and but-2-ynoyl chloride (14 mg, 0.14 mmol) was added dropwise, reacted at 0°C for 4 h, and the reaction was complete after TLC spot plate monitoring. The crude product was purified by Pre-HPLC to obtain the final product XD37 (33 mg, yield 57%).

[0438] LC-MS: m / z=474.1[M+H] +

[0439] 1H NMR (400MHz, DMSO) δ10.86(s,1H),8.19(d,J=2.0Hz,1H),7.87(d,J=2.5Hz,1H),7.73(d,J=8.9Hz,1H),7.61–7.52(m,2H),4.93(dd,J=10.3,5.1Hz,1H) ,2.21(ddd,J=14.5,10.2,4.5Hz,1H),2.07(s,3H),1.99(ddd,J=14.3,9.2, 5.2Hz,1H),1.13–0.99(m,1H),0.67(d,J=6.6Hz,3H),0.57(d,J=6.5Hz,3H). 13 C NMR(101MHz,DMSO)δ170.55,167.63,151.74,151.14,135.69,132.87,130.18,129.38,128.17,1 24.09,123.78,117.86,114.72,108.05,84.99,76.29,58.29,38.55,24.61,22.81,21.18,3.68.

[0440] Embodiment 38

[0441]

[0442] The reactant S3 (50 mg, 0.12 mmol) was dissolved in 4 mL of dichloromethane, stirred under ice bath and bromoacetyl chloride (21 mg, 0.14 mmol) was added dropwise, reacted at 0°C for 4 h, and the reaction was complete after TLC spot plate monitoring. The crude product was purified by Pre-HPLC to obtain the final product XD38 (48 mg, yield 74%).

[0443] LC-MS: m / z = 528.0 [M+H] +

[0444] 1 H NMR (400MHz, DMSO) δ10.75–10.58(m,1H),8.22(d,J=1.9Hz,1H),7.89(d,J=2.6Hz,1H),7.76(d,J=8.9Hz,1H),7.61(d,J=2.6Hz,1H),7.53(ddd,J=9.0 ,3.8,2.0Hz,1H),4.95(dd,J=10.2,5.2Hz,1H),4.36–4.08(m,2H),2.25–1. 97(m,2H),1.14–1.00(m,1H),0.68(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H).13 C NMR (101MHz, DMSO) δ170.54,165.49,165.34,162.78,151.75,148.96,135.83,132.95,130.24,129. 38,124.12,123.81,117.78,114.88,58.32,44.05,38.57,36.25,31.23,30.86,24.62,22.81,21.23.

[0445] Embodiment 39

[0446]

[0447] The reactant S3 (50 mg, 0.12 mmol) was dissolved in 4 mL of dichloromethane, stirred under ice bath and 2-chloro-2-fluoroacetyl chloride (18 mg, 0.14 mmol) was added dropwise, reacted at 0°C for 4 h, and the reaction was complete after TLC spot plate monitoring. The crude product was purified by Pre-HPLC to obtain the final product XD39 (35 mg, yield 57%).

[0448] LC-MS: m / z=502.1[M+H] +

[0449] 1 H NMR (400MHz, DMSO) δ11.05(s,1H),8.24(t,J=2.4Hz,1H),7.87(d,J=2.6Hz,1H),7.80(d,J=9.0Hz,1H),7.65(dt,J=9.0,2.0Hz,1H),7.61(d,J=2. 6Hz,1H),7.08–6.89(m,1H),4.95(dd,J=10.3,5.2Hz,1H),2.27–1.98(m, 2H), 1.08 (t, J=8.8Hz, 1H), 0.67 (d, J=6.6Hz, 3H), 0.58 (d, J=6.5Hz, 3H). 13 CNMR(101MHz,DMSO)δ170.51,162.69,162.46,151.73,138.04,134.54,132.96,130.27,129.92,1 24.14,123.83,118.39,117.80,114.98,108.72,95.41,92.91,58.36,38.58,24.63,22.78,21.19.

[0450] Embodiment 40

[0451]

[0452] The reactant S3 (50 mg, 0.12 mmol) was dissolved in 4 mL of dichloromethane, stirred under ice bath and 2-dichloroacetyl chloride (20 mg, 0.14 mmol) was added dropwise, reacted at 0°C for 4 h, and the reaction was complete after TLC spot plate monitoring. The crude product was purified by Pre-HPLC to obtain the final product XD40 (44 mg, yield 69%).

[0453] LC-MS: m / z=518.0 [M+H] +

[0454] 1 H NMR (400MHz, DMSO) δ11.10(s,1H),8.23(d,J=2.0Hz,1H),7.88(d,J=2.5Hz,1H),7.81(d,J=8.9Hz,1H),7.64–7.56(m,2H),6.7 3(s,1H),4.95(dd,J=10.2,5.2Hz,1H),2.29–1.95(m,2H),1.07(q,J=7.2Hz,1H),0.67(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H). 13 C NMR(101MHz,DMSO)δ170.51,162.47,151.73,138.08,134.76,132.97,130.29,129.90,12 4.17,123.84,118.17,117.80,115.09,108.42,67.81,58.37,38.58,24.63,22.79,21.20.

[0455] Embodiment 41

[0456]

[0457] The reactant S3 (50 mg, 0.12 mmol) was dissolved in 4 mL of dichloromethane, stirred under ice bath and 2-dichloroacetyl chloride (22 mg, 0.14 mmol) was added dropwise, reacted at 0°C for 4 h, and the reaction was complete after TLC spot plate monitoring. The crude product was purified by Pre-HPLC to obtain the final product XD41 (47 mg, yield 72%).

[0458] LC-MS: m / z=534.1[M+H] +

[0459] 1H NMR (400MHz, DMSO) δ10.81(s,1H),8.32(d,J=2.0Hz,1H),7.86(d,J=2.6Hz,1H),7.7 2(d,J=8.9Hz,1H),7.56(dt,J=5.5,2.4Hz,2H),7.27(d,J=15.4Hz,1H),6.76(d,J=15 .4Hz,1H),4.93(dd,J=10.2,5.2Hz,1H),4.24(q,J=7.1Hz,2H),2.26–1.94(m,2H),1. 28(t,J=7.1Hz,3H),1.11–1.01(m,1H),0.67(d,J=6.6Hz,3H),0.58(d,J=6.5Hz,3H). 13 C NMR (101MHz, DMSO) δ170.76,165.41,161.84,151.68,149.57,138.13,135.26,132.54,130.13,130.03,12 9.93,124.04,123.70,118.75,117.49,114.53,108.58,61.32,58.11,38.60,24.64,22.84,21.24,14.50.

[0460] Biological activity test

[0461] Example 1 Binding ability of compounds to nsp8 nonstructural protein

[0462] The experimental results are as follows Figure 1 As shown, Figure 1 The mass spectrometry results of A show the mass spectrum of nsp8 non-structural protein. Figure 1 B shows the mass spectrometry results after compound XD9 covalently binds to nsp8 protein. The increase in molecular weight indicates that the compound binds to the protein.

[0463] Example 2 RdRp complex activity test method

[0464] Experimental method: According to the literature method (Science. 2020, 368, 1499-1504.), 5'-FAM-labeled RNA double-stranded (template sequence is 5'-UUUUUUUUUUAUAACUUAAUCUCACAUAGC-3', primer sequence is 5'-FAM-GCUAUGUGAGAUUAAGUUAU-3') and recombinant protein SARS-CoV-2RdRp were prepared.

[0465] Prepare 4X reaction buffer (4XRB) containing 80mM Tris (pH 8.0), 40mM KCl, 24mM MgCl2 and 0.04% Triton-X100 with DEPC water. During the test, SARS-CoV-2RdRp and RNA duplex were diluted to 4μM and 12μM respectively using 4XRB. 4.5μL RdRp (4μM), 4.5μL RNA duplex (12μM) and 9μL of different concentrations of the test compound were added to each reaction. The reaction system was then incubated at room temperature in the dark for 1 hour. Then 2μL ATP (100mM) was added to each reaction, and the reactants were placed in a 37°C water bath for 1 hour. Finally, 40μL of stop solution (94% formamide, 30mM EDTA) was added to terminate the reaction. Take 10μL of the reaction product and mix it with 2μL 6X DNA loading buffer (Thermo Scientific). Prepare a polyacrylamide gel containing 15% urea. Load 10 μL of sample. Electrophoresis was performed at 150 V for 90 minutes, and imaging was performed using a Bio-Rad gel imager. The above reagents were purchased from Sigma-Aldrich, RNA was synthesized by Sangon Biotech (Shanghai) Co., Ltd., and the gel imager model was Bio-Rad ChemiDoc XRS+.

[0466] The synthesized RNA was heated at 55°C for 5 minutes to anneal, cooled at room temperature and dissolved in TE buffer to 100uM. 4X reaction buffer (4X RB) containing 80mM Tris (pH 8.0), 40mM KCl, 24mM MgCl2 and 0.04% Triton-X100 was prepared with DEPC water. During the test, 4XRB was diluted to 1X with DEPC water. 14uL of different concentrations of the test compound, 14ul SARS-CoV-2RdRp (800nM), 14uL ATP (100mM) and 14uL RNA (400nM) were added to the test system in sequence. The required concentrations for the above components were obtained by diluting 1X RB. Then place in a 37°C water bath for 20min. Finally, 14uL dsDNADye was added, mixed by pipetting, and incubated at room temperature in the dark for 5min. Take 20ul of the reactant and add it to a 384-well plate, with 3 replicates for each reaction. Finally, the reading was performed using an ELISA reader. The excitation wavelength and emission wavelength were set to 485 nm and 535 nm, respectively. dsDNA Dye was purchased from Promega, catalog number E2670. The RNA substrate was synthesized by Sangon Biotech (Shanghai) Co., Ltd., and the sequence is as follows: 5'-UUUUUUUUUUUUUUUUUUUUCACAUAGCUUUUUGCUAUGUG-3'

[0467] The remaining reagents were purchased from Sigma-Aldrich. The 384-well plate was Corning ELISA plate No. 3575, and the ELISA reader was PerkinElmer EnVision 2104 Multilabel Reader.

[0468] Experimental results: The compounds of the present invention have the effect of significantly inhibiting the activity of the RdRp complex, thereby affecting the RNA virus replication process and inhibiting the proliferation of the virus.

[0469] The results showed that the compound had a significant covalent inhibitory effect on nsp8 in the rdrp complex as verified by covalent mass spectrometry, and the enzyme activity experiment further verified that the compound could inhibit the extension effect of the rdrp complex on RNA. Therefore, the compound described in the present application can affect the activity of the complex by affecting the binding of nsp8 and nsp7 / 12.

[0470] A: IC 50 <10μM; B:10μM<IC 50 <50μM; C:50μM<IC 50 <100μM;D:

[0471] 100μM<IC 50

[0472]

[0473]

[0474] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof; in, R1, R2, R3 and R4 are each independently selected from the group consisting of H, -NH2, halogen, -NO2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O and S, -C(=O )R6, -OC(=O)R6, -C(=O)OR6, -OR6, -SR6, -S(=O)R6, -S(=O)2R6, -S(=O)2N(R6)2, -N(R6)2, -C(=O)N(R6)2, -NR6-C(=O)R6, -NR6-C(=O)OR6, -NR6-S(=O)2-R6, -C1-C6 alkylene-N(R6)2, -C1-C6 alkylene-OR6, -C2-C6 alkenylene-OR6, -O-C1-C6 alkylene-N(R6)2; Y is selected from the group consisting of C, N; When Y is N, R3 does not exist; R5 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S; The substitution refers to substitution by one or more substituents selected from the group consisting of halogen, -NH2, -OH, C1-C6 alkoxy, -CN, -NO2, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy C1-C6 alkyl, C3-C10 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, C2-C6 acyl, -C(=O)OR6; Wherein, the R6 is selected from the following group: H, guanidinyl, C1-C6 alkyl, saturated or partially unsaturated C3-C6 cycloalkyl, saturated or partially unsaturated C3-C10 heterocycloalkyl; A is selected from the following group: absent, -OR 10 、-C1-C6 alkyl-R 10 、-N(R 10 )2, -C1-C6 alkylene-N(R 10 )2. -C(=O)-R 10 、-NH-C(=O)-R 10 、-C(=O)-N(R 10 )2. -SO2-R 10 、-NH-SO2-R 10 、-SO2-N(R 10 )2; R 10 Each is independently selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; or two R 10 The substituted or unsubstituted 5-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, or the substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution by a substituent selected from the group consisting of: -OH, -CN, -NH2, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(=O)-C1-C6 alkyl, -C(=O)-halogenated C1-C6 alkyl, -C(=O)-C2-C6 alkenyl, -C(=O)-C2-C6 alkynyl, -NH-C1-C C1-C6 alkyl, -NH-halo-C1-C6 alkyl, -NH-C2-C6 alkenyl, -NH-C2-C6 alkynyl, -N(C1-C6 alkyl)2, -N(halo-C1-C6 alkyl)2, -N(C2-C6 alkenyl)2, -N(C2-C6 alkynyl)2, -NH-C(=O)-C1-C6 alkyl, -NH-C(=O)-halo-C1-C6 alkyl, -COOH, -C(=O)-O-C1-C6 alkyl, -C(=O)-O-halo-C1-C6 alkyl, -C(=O)-O-C2-C6 alkenyl, C(=O)-O-C2-C6 alkynyl; B is selected from the following groups: H, halogen, -NO2, C1-C6 alkyl, halo-substituted C1-C6 alkyl, saturated or partially unsaturated C3-C10 heterocycloalkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution by one or more substituents selected from the following groups: halogen, -NH2, -OH, oxo, -CN, -NO2, C1-C6 alkyl, halo-substituted C1-C6 alkyl, hydroxy C1-C6 alkyl, C1-C6 alkoxy, C3-C10 cycloalkyl, C3-C8 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, -COOH, C2-C6 acyl.

2. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, characterized in that: R1, R2, R3 and R4 are each independently selected from the group consisting of H, -NH2, halogen, -NO2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted C2-C6 alkyl containing 1-3 heteroatoms selected from N, O and S Heteroatom-substituted or unsubstituted 5-7 membered heteroaryl, -C(=O)R6, -OC(=O)R6, -C(=O)OR6, -OR6, -SR6, -S(=O)R6, -S(=O)2R6, -N(R6)2, -C(=O)N(R6)2, -NR6-C(=O)R6, -NR6-C(=O)OR6, -C1-C6 alkylene-N(R6)2, -C1-C6 alkylene-OR6, -C2-C6 alkenylene-OR6; Y is C; R5 is selected from the following group: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S, -C(=O)R6, -OR6; The substitution refers to substitution by one or more substituents selected from the group consisting of halogen, -NH2, -OH, C1-C6 alkoxy, -CN, -NO2, C1-C6 alkyl, C3-C10 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, -C(=O)OR6; A is selected from the following group: absent, -OR 10 、-C1-C6 alkyl-R 10 、-N(R 10 )2. -C(=O)-R 10 、-NH-C(=O)-R 10 、-C(=O)-N(R 10 )2. -SO2-R 10 、-SO2-N(R 10 )2; R 10 Each is independently selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; or two R 10 connected to form a substituted or unsubstituted 5-7 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution with a substituent selected from the following group: -OH, -CN, -NH2, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(=O)-C1-C6 alkyl, -C(=O)-halogenated C1-C6 alkyl, - C(=O)-C2-C6 alkenyl, -C(=O)-C2-C6 alkynyl, -NH-C1-C6 alkyl, -NH-haloC1-C6 alkyl, -NH-C2-C6 alkenyl, -NH-C2-C6 alkynyl, -N(C1-C6 alkyl)2, -NH-C(=O)-C1-C6 alkyl, -NH-C(=O)-haloC1-C6 alkyl, -COOH, -C(=O)-O-C1-C6 alkyl, -C(=O)-O-haloC1-C6 alkyl.

3. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, characterized in that: R1, R2, R3 and R4 are each independently selected from the group consisting of H, -NH2, halogen, -NO2, substituted or unsubstituted C1-C6 alkyl, -OR6; R5 is selected from the following groups: H, substituted or unsubstituted C1-C6 alkyl, saturated or partially unsaturated substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S; A is selected from the following group: absent, -OR 10 、-C1-C6 alkyl-R 10 、-N(R 10 )2. -C(=O)-R 10 、-NH-C(=O)-R 10 、-C(=O)-N(R 10 )2. -SO2-R 10 、-SO2-N(R 10 )2; R 10 Each is independently selected from the following group: H, halogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl; or two R 10 connected to form a substituted or unsubstituted 5-7 membered heterocycloalkyl group containing 1-3 heteroatoms selected from N, O or S, or a substituted or unsubstituted 5-7 membered heteroaryl group containing 1-3 heteroatoms selected from N, O or S; the substitution refers to substitution by a substituent selected from the following group: -OH, -CN, -NH2, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, -C(=O)-C1-C6 alkyl, -C(=O)-halogenated C1-C6 alkyl, -C(=O)-C2-C6 alkenyl, -C(=O)-C2-C6 alkynyl, -NH-C(=O)-C1-C6 alkyl, -NH-C(=O)-halogenated C1-C6 alkyl, -COOH, -C(=O)-O-C1-C6 alkyl.

4. The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, characterized in that: The compound of formula I is a compound of formula II, Wherein, R1, R2, R3, R4, R5 and B are defined as in claim 1; Having a structure selected from the group consisting of: X is each independently selected from the following group: n is selected from the following group: 0, 1, 2, 3.

5. The compound of claim 4, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, characterized in that: R1 and R2 are each independently selected from the group consisting of H, -OH, halogen, -NO2, C1-C6 alkyl, C1-C6 haloalkyl, -O-C1-C3 alkyl; R3 and R4 are each independently selected from: H, -OH, halogen, -NO2, C1-C6 alkyl, R5 is selected from the following group: H, -OH, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, saturated or partially unsaturated substituted or unsubstituted C3-C10 heterocycloalkyl containing 1-3 heteroatoms selected from N, O and S; the substitution refers to substitution by one or more substituents selected from the following group: halogen, -NH2, -OH, C1-C6 alkoxy, -CN, -NO2, C1-C6 alkyl, halo C1-C6 alkyl, hydroxy C1-C6 alkyl, C3-C10 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, C2-C6 acyl, -C(=O)OR6; Having a structure selected from the group consisting of: X is selected from the following group: B is selected from the following groups: H, halogen, -NO2, -CF3, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; the substitution refers to replacement by one or more substituents selected from the following groups: halogen, -NH2, -OH, oxo, -CN, -NO2, C1-C6 alkyl, halogenated C1-C6 alkyl, hydroxy C1-C6 alkyl, C1-C6 alkoxy, C3-C10 cycloalkyl, C3-C8 membered heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, -COOH, C2-C6 acyl.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, characterized in that: The compound is selected from the group consisting of:

7. A method for preparing a compound of formula I, comprising the following steps: In an inert solvent and at a certain temperature, compound I-1 reacts with compound I-2 to prepare a compound of formula I.

8. A pharmaceutical composition comprising: (1) The compound of claim 1, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof; and (2) A pharmaceutically acceptable carrier.

9. A use of the compound of claim 1, or a pharmaceutically acceptable salt, tautomer, enantiomer, diastereomer, racemate, hydrate, ester, solvate, metabolic precursor or prodrug thereof, characterized in that: Used for preparing medicines or pharmaceutical compositions for preventing and / or treating diseases caused by viral infections.

10. The use according to claim 9, characterized in that The disease is selected from the group consisting of coronavirus, calicivirus, and picornavirus infection.

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