A cyclopeptide viral protease inhibitor, its preparation and use in antiviral medicaments

Abstract

The application belongs to the field of pharmaceutical chemical industry and relates to a cyclic peptide compound shown as formula M or a stereoisomer, a tautomer or a mixture thereof of the compound, a pharmaceutically acceptable salt, a polymorph, a co-crystal or a solvate of the compound, or a stable isotope derivative, a metabolite or a prodrug of the compound. The application also relates to a method for synthesizing the cyclic peptide compound and an intermediate. The provided cyclic peptide compound has a strong inhibitory effect on a coronavirus 3CL protease and a small RNA virus (for example, an enterovirus EV71 virus) 3C protease. After cell level activity testing, the provided cyclic peptide compound has an obvious inhibitory effect on EV71 virus and coronavirus and the like, indicating that the cyclic peptide compound shown as formula M can produce a viral inhibitory effect at the cell level by inhibiting viral protease activity, and has a good application prospect as an antiviral drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemical industry, and relates to a kind of cyclic peptide compounds, and its application as viral protease inhibitor, in the preparation of antiviral drugs. Specifically, the present application relates to a kind of cyclic peptide compounds, the stereoisomer, tautomer or mixture thereof of the compound, the pharmaceutically acceptable salt, polymorph, co-crystal or solvate of the compound, or, the stable isotope derivative, metabolite or prodrug of the compound, and their application in inhibiting viral protease, preparing for preventing and / or treating the infection disease caused by the virus (mainly including but not limited to small RNA virus and coronavirus) in which protease plays an important role in viral life cycle. The present application also relates to the method and intermediate for synthesizing the cyclic peptide compounds. BACKGROUND

[0002] Small RNA viruses and coronaviruses belong to single positive strand RNA viruses. The small RNA virus family mainly includes enterovirus (CV, PV, EV71 virus, etc.), human rhinovirus (HRV) and hepatitis A virus (HAV) etc. EV71 virus and coxsackievirus infection can cause herpangina and hand-foot-mouth disease in children, and can be life-threatening in severe cases. Poliovirus (PV) and hepatitis A virus (HAV) can cause poliomyelitis (usually manifested as infantile paralysis) and hepatitis A, respectively. Human rhinovirus is a kind of rhinovirus (RhV), which is the virus with the most serotypes among human viruses, and is the main pathogen causing common cold. The virus is the culprit of acute respiratory disease, and nearly half of the acute respiratory disease infections are caused by rhinovirus infection.

[0003] Coronaviruses belong to the order of Nidovirales, the family of Coronaviridae and the genus of Coronavirus in systematic classification. There are currently 7 known coronaviruses that can infect humans.

[0004] In the process of viral replication, the genome of some viruses first encodes a large polyprotein precursor, and then the functional proteins are produced by proteolysis of the polyprotein, which is mainly completed by proteases. The precursor protein produced by most single-stranded RNA viruses can only form functional proteins after being hydrolyzed by 3C or 3CL protease, and then the subsequent replication and packaging can be carried out, so 3C and 3CL proteases are the core proteases for hydrolysis of the precursor polyprotein of single-stranded RNA viruses, and play a crucial role in the process of single-stranded RNA virus replication. For example, small RNA viruses first encode a large polyprotein precursor, and then the hydrolysis of the polyprotein precursor is completed by 3C protease, and the coronavirus first encodes two polyproteins (pp1a and pp1ab), and then the functional proteins are also produced by hydrolysis of the polyprotein, and the hydrolysis process is mainly completed by 3CL protease. In the Flavivirus genus, the hydrolysis of the polyprotein is mainly completed by non-structural protein NS3 / 4A. In the life cycle of non-single-stranded RNA viruses such as human immunodeficiency virus (HIV), similar proteases are also involved.

[0005] Because there is no similar protease in the human body with such structure and function, inhibiting the catalytic function of viral proteases can effectively inhibit the cleavage of viral precursor proteins and block viral replication. 3C or 3CL protease is an important target for the study of single-stranded RNA virus drug treatment. Although single-stranded RNA virus genes have diversity, the 3C and 3CL protease substrate binding sites are highly conserved and have similar catalytic mechanisms, which are highly similar proteases in single-stranded RNA viruses. Therefore, broad-spectrum anti-single-stranded RNA virus inhibitors targeting 3C and 3CL proteases have received widespread attention. 3C and 3CL proteases belong to cysteine proteases and have a highly conserved three-dimensional structure. Although 3C and 3CL proteases have low sequence homology, sequence analysis based on structural basis found that the two types of proteases have highly conserved Gly-X-Cys-Gly-Gly-Gly / Ser sequence structures, and the His-Cys in the catalytic triad of 3C protease is almost completely identical to that of 3CL protease, indicating that the binding sites of the two types of proteases are highly conserved and have similar catalytic mechanisms.

[0006] Rupintrivir (AG7088, Rupintrivir) was originally developed by Agouron Pharmaceuticals, Inc. It is a specific inhibitor of human rhinovirus and is a peptide drug. It has a similar spatial configuration to the 3C protease substrate and thus competes with the substrate for binding to the 3C protease, exerting an inhibitory effect on the enzyme. Rupintrivir inhibits the replication of 48 different HRV serotypes in H1-HeLA and MRC-5 cell protection assays with an average EC50 of 0.023 μM. Rupintrivir has immunomodulatory effects. There are reports that Rupintrivir has therapeutic effects on EV71 infected animals. There are also reports that Rupintrivir can inhibit SARS coronavirus by inhibiting the 3CL protease of SARS coronavirus. The above facts show that a compound can have broad-spectrum antiviral effects on 3C protease or 3CL protease. SUMMARY

[0007] The cyclic peptide compounds of general formula M are potent viral protease inhibitors. The compounds of general formula M are useful for inhibiting viral proteases and for preventing and / or treating infectious diseases caused by viruses in which proteases play an important role in the viral life cycle, including but not limited to, picornaviruses and coronaviruses. It is another object of the present application to provide a process for the preparation of cyclic peptide protease inhibitors described by general formula M and their synthetic intermediates, for the synthesis of compounds of general formula M and their intermediates.

[0008] In one aspect, the present application provides a cyclic peptide compound of formula M, a stereoisomer, a tautomer, or a mixture thereof, a pharmaceutically acceptable salt, a polymorph, a co-crystal, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound,

[0009]

[0010] In formula M, R1is selected from wherein R' is C1-C6alkyl or C3-C6cycloalkyl,

[0011] R2is C1-C6alkyl or C3-C6cycloalkyl, or aryl substituted with one or more (e.g., 2, 3, 4) substituents selected from the group consisting of fluorine, chlorine, bromine, iodine, C1-C6alkyl, C1-C6alkoxy, cyano, nitro;

[0012] Y is selected from C, N, O; n + m = 4, 5, 6, 7, or 8;

[0013] Ring A is selected from a substituted or unsubstituted five- or six-membered aryl group, a substituted or unsubstituted five- or six-membered heteroaryl group. In certain embodiments, in formula M, wherein R' is C1-C3 alkyl or C3-C6 cycloalkyl,

[0014] R2 is C1-C6 alkyl or C3-C6 cycloalkyl, or one or more (e.g. 2, 3, 4) fluorine substituted phenyl rings,

[0015] Y is selected from C, N, O; n+m = 5, 6 or 7,

[0016] A ring is a phenyl ring or a five-membered heteroaryl, which preferably contains 1, 2 or 3 nitrogen atoms and / or 1, 2 or 3 oxygen atoms.

[0017] In certain embodiments, in formula M, R1 is selected from wherein R' is methyl, ethyl or cyclopropyl,

[0018] R2 is selected from cyclopropyl, cyclohexyl, isopropyl, 4-fluorophenyl, 3- fluorophenyl, 3,4-difluorophenyl,

[0019] Y is selected from, C, O; n+m = 5, 6, 7 or 8

[0020] A ring is selected from the following five-membered heteroaryl

[0021]

[0022] In certain embodiments, R1 is selected from

[0023] In certain embodiments, R' is C1-C4 alkyl, such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl.

[0024] In certain embodiments, R' is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0025] In certain embodiments, R1 is selected from wherein R' is selected from methyl, ethyl, cyclopropyl.

[0026] In certain embodiments, R2 is phenyl substituted with one or more (e.g. 2, 3, 4) substituents selected from fluorine, chlorine, bromine, iodine, C1-C6 alkyl (e.g. C1-C4 alkyl or C1-C3 alkyl), C1-C6 alkoxy (e.g. methoxy or ethoxy), cyano, nitro.

[0027] In certain embodiments, R2 is phenyl substituted with one or more (e.g. 2, 3, 4) substituents selected from fluorine, chlorine, bromine.

[0028] In certain embodiments, R2 is phenyl substituted with one or more (e.g. 2, 3, 4) fluorine.

[0029] In certain embodiments, R2is C3-C6cycloalkyl.

[0030] In certain embodiments, R2is selected from cyclopropyl, cyclohexyl, isopropyl, 4- fluorophenyl, 3-fluorophenyl, 3,4-difluorophenyl.

[0031] In certain embodiments, Y is C or O.

[0032] In certain embodiments, n+m = 5, 6 or 7.

[0033] In certain embodiments, the A ring is a phenyl ring or a five-membered heteroaryl group containing 1, 2 or 3 nitrogen atoms and / or 1, 2 or 3 oxygen atoms.

[0034] In certain embodiments, the A ring is selected from the following five-membered heteroaryl groups

[0035]

[0036] The compounds of the present application can have a structure selected from the following:

[0037]

[0038]

[0039] In another aspect, the present application provides a method of preparing a compound of the present application, a stereoisomer, a tautomer, or a mixture thereof, a pharmaceutically acceptable salt, a polymorph, a co-crystal, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, the method comprising: synthesizing a compound of Formula 5 (Intermediate 5) by the following reaction scheme:

[0040]

[0041] wherein A, R2, m and n are as described above, and reaction conditions a and b are inorganic bases including, but not limited to, sodium hydroxide, sodium carbonate, lithium hydroxide.

[0042] In certain embodiments, the compounds of the present application have a structure as shown in Formula 6, and the method of synthesis can employ the following reaction scheme:

[0043]

[0044] The compound of Formula 5 can be prepared using the route described above.

[0045] In certain embodiments, the compounds of the present application have a structure as shown in Formula 7, and the method of synthesis can employ the following reaction scheme:

[0046] In certain embodiments, the compounds of the present application have a structure as shown in Formula 6, and the method of synthesis can employ the following reaction scheme:

[0047] wherein the reaction condition c is an inorganic base, including but not limited to sodium hydroxide, sodium carbonate, lithium hydroxide.

[0048] The compound of formula 5 can be prepared by following routes as described above.

[0049] In certain embodiments, the compound of formula 2 has a structure shown in formula 2-1, and the compound of formula 2-1 is prepared by following reaction route:

[0050]

[0051] The reaction can be carried out in the presence of Cu(I), for example, in the presence of ascorbic acid and copper sulfate pentahydrate.

[0052] In certain embodiments, the compound of formula 2 has a structure shown in formula 2-2, and the compound of formula 2-2 is prepared by following reaction route:

[0053]

[0054] The reaction can be carried out in the presence of an inorganic base, for example, sodium bicarbonate.

[0055] In certain embodiments, the compound of formula 2 has a structure shown in formula 2-3, and the compound of formula 2-3 is prepared by following reaction route:

[0056]

[0057] The present application also provides a pharmaceutical composition comprising at least one compound of the present application, a stereoisomer, a tautomer, or a mixture thereof, a pharmaceutically acceptable salt, a polymorph, a co-crystal, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound; optionally, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, carrier, vehicle, or adjuvant.

[0058] Optionally, the pharmaceutical composition further comprises an EV71 antiviral agent; in certain embodiments, the EV71 antiviral agent is an antiviral agent selected from 3D protease inhibitors and VP1 protein inhibitors.

[0059] In certain embodiments, the pharmaceutical composition is used for preventing / treating a disease associated with viral infection in a subject, the virus is selected from the group consisting of picornavirus (e.g. enterovirus, human rhinovirus (HRV) and hepatitis A virus (HAV)), and coronavirus. Among them, the enterovirus includes but is not limited to enterovirus 71 (EV71), poliovirus, coxsackievirus A, coxsackievirus B, and the coronavirus includes but is not limited to SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV.

[0060] SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and

[0061] MERS-CoV. In certain embodiments, the pharmaceutical composition is used for preventing / treating a disease associated with enterovirus 71 (EV71) infection or a disease associated with SARS-CoV-2 infection in a subject. In certain embodiments, the subject is a mammal, such as a bovine, equine, porcine, canine, feline, rodent, primate. Among them, the particularly preferred subject is a human.

[0062] The present application also provides the use of a compound of the present application, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt, a polymorph, a co-crystal or a solvate of the compound, or a stable isotope derivative, a metabolite or a prodrug of the compound in the preparation of a medicament, which is a viral protease inhibitor.

[0063] In certain embodiments, the virus is selected from the group consisting of picornavirus (e.g. enterovirus (e.g. coxsackievirus (CV), poliovirus (PV), enterovirus 71 type), human rhinovirus (HRV) and hepatitis A virus (HAV)), and coronavirus (e.g. SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV).

[0064] In certain embodiments, the protease is 3C / 3CL protease.

[0065] The present application also provides the use of a compound of the present application, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt, a polymorph, a co-crystal or a solvate of the compound, or a stable isotope derivative, a metabolite or a prodrug of the compound in the preparation of a medicament, which is an antiviral drug;

[0066] Preferably, the virus against which the antiviral drug is directed is selected from the group consisting of picornaviruses (e.g. enterovirus genus (e.g. coxsackievirus (CV), poliovirus (PV), enterovirus 71), human rhinovirus genus (HRV) and hepatitis A virus genus (HAV)), and coronaviruses (e.g. SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV).

[0067] The present application also provides the use of a compound of the present application, a stereoisomer, a tautomer, or a mixture thereof, a pharmaceutically acceptable salt, a polymorph, a co-crystal, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, in the preparation of a medicament for preventing / treating a disease associated with viral infection in a subject, the virus being selected from the group consisting of picornaviruses (e.g. enterovirus genus, human rhinovirus genus (HRV) and hepatitis A virus genus (HAV)), and coronaviruses. Among them, the enterovirus includes but is not limited to enterovirus 71 (EV71), poliovirus, coxsackievirus A, coxsackievirus B, and the coronavirus includes but is not limited to SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV. In certain embodiments, the subject is a mammal, such as a bovine, equine, porcine, canine, feline, rodent, primate. Among them, a particularly preferred subject is a human.

[0068] In another aspect, the present application provides a method of preventing / treating a disease associated with viral infection in a subject, comprising the step of administering to the subject a prophylactically / treatingly effective amount of a compound of the present application, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a polymorph, a co-crystal, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, or a pharmaceutical composition of the present application, the virus is selected from the group consisting of a picornavirus (e.g., enterovirus (e.g., coxsackievirus (CV), poliovirus (PV), enterovirus 71), human rhinovirus (HRV), and hepatitis A virus (HAV)), and a coronavirus (e.g., SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, and MERS-CoV). In certain embodiments, the subject is a mammal, such as a bovine, equine, porcine, canine, feline, rodent, primate. Among them, a particularly preferred subject is a human. In yet another aspect, the present application also provides the use of a compound having the structure shown in Formula 5, a stereoisomer, a tautomer, or a mixture thereof of the compound, a pharmaceutically acceptable salt, a polymorph, a co-crystal, or a solvate of the compound, or a stable isotopic derivative, a metabolite, or a prodrug of the compound, as an intermediate for preparing the cyclic peptide compound of the present application:

[0069]

[0070] wherein A, R2, m and n are as described above.

[0071] Definitions of Terms

[0072] In the present application, unless otherwise indicated, the scientific and technical terms used herein have the meanings that would be generally understood by one of ordinary skill in the art, however, for better understanding of the present application, the definitions of some terms are provided below. When the definitions and explanations of the terms provided in the present application are inconsistent with the meanings commonly understood by one of ordinary skill in the art, the definitions and explanations of the terms provided in the present application shall prevail. When referring to each example, (R) or (S) is used to indicate the absolute configuration of an asymmetric center, which indicates the description for the entire compound rather than the description for a single substituent.

[0073] As used herein, "C1-C6alkyl" means straight or branched chain alkyl groups having from 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, t-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl, and the like; C1-C3alkyl can be similarly understood. Preferred is C1-C3alkyl.

[0074] As used herein, "C1-C6alkoxy" means a straight or branched chain alkoxy group having from 1 to 6 carbon atoms, for example methoxy, ethoxy, propyloxy, isopropoxy, n-butyloxy, sec-butyloxy, t-butyloxy, pentyloxy, 2-pentyloxy, isopentyloxy, neopentyloxy, hexyloxy, 2-hexyloxy, 3-oxahexyloxy, and the like; C1-C3alkoxy is also to be similarly construed. Preferred is C1-C3alkoxy.

[0075] As used herein, the term "aryl" means an aromatic monocyclic ring system containing 6 carbon atoms, or an aromatic bicyclic ring system containing 10 atoms, for example phenyl and naphthyl ring systems.

[0076] As used herein, the term "heteroaryl" alone or in combination with another substituent means a monovalent radical derived by removal of a hydrogen from a five-, six-, or seven-membered unsaturated heterocyclic ring having, for example, one, two, or three heteroatoms selected from N, O, S. Representative examples of suitable heteroaryl groups are: thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, isoxazolyl, 1,2,3-triazolyl.

[0077] As used herein, "C3-C6cycloalkyl" means a saturated or partially saturated and non-aromatic monocyclic ring group containing from 3 to 6 ring atoms, including "3-6 membered saturated cycloalkyl" and "3-6 membered partially saturated cycloalkyl", for example "5-6 membered cycloalkyl", "5-6 membered saturated cycloalkyl", and the like. Examples include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cyclohexenyl, and the like.

[0078] The term "stereoisomer" as used herein denotes isomers that arise due to the presence of at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, it is possible to produce racemic mixtures, single enantiomers, mixtures of diastereomers, and individual diastereomers. Particular individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present application can exist as mixtures of two or more structurally distinct forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, and the like. It is to be understood that the scope of the present application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%).

[0079] The present application encompasses all possible crystalline forms or polymorphs of the compounds of the present application, which can be a single polymorph or a mixture of more than one polymorph in any proportion.

[0080] It should also be understood that certain compounds of the present application can exist in free form for treatment, or where appropriate, as a pharmaceutically acceptable derivative thereof. In this context, a pharmaceutically acceptable derivative includes, but is not limited to, a pharmaceutically acceptable salt, solvate, metabolite or prodrug, which upon administration to a patient in need thereof, is capable of being converted into the compound of the present application or a metabolite or residue thereof.

[0081] The term "pharmaceutically acceptable salt" as used herein refers to salts of the compounds of Formula (M) which are suitable for use in normal medical therapy and which are not toxic, injurious, allergenic, etc. to the tissues of humans and animals at the dosages administered. They are generally water or oil-soluble or dispersible, and are effective for their use. This term includes both pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0082] The term "pharmaceutically acceptable acid addition salt" refers to those salts which retain the biological activity of the free base and which are not biologically or otherwise undesirable, formed with inorganic acids such as sulphuric acid, nitric acid, phosphoric acid, hydrochloric acid, sulphamic acid and the like, and organic acids such as acetic acid, trifluoroacetic acid, trichloroacetic acid, cinnamic acid, citric acid, maleic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzoic acid, benzenesulphonic acid, glycolic acid, malic acid, lactic acid, malonic acid, oxalic acid, nicotinic acid, succinic acid, salicylic acid, stearic acid, tartaric acid, p-aminobenzoic acid, trimethyl-benzenesulphonic acid, p-toluenesulphonic acid, mandelic acid, embonic acid, picric acid, propionic acid and the like.

[0083] The term "pharmaceutically acceptable base addition salt" refers to those salts which retain the biological activity of the free acid and which are not biologically or otherwise undesirable, formed with inorganic bases, such as ammonia or ammonium hydroxide or metal cations, such as sodium, magnesium, calcium, potassium, aluminum, and the like, and particularly preferred are the ammonium, potassium, sodium, calcium, and magnesium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, quaternary ammonium compounds, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as methylamine, dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine, tripropylamine, isopropylamine, tributylamine, ethanolamine, diethanolamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine.

[0084] The term "solvate" as used herein refers to a compound associated with a solvent molecule. The solvent can be water or an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, etc.) or the like.

[0085] Metabolites of the compounds of the application, i.e., species derived from the compounds of the application in vivo, are also included within the scope of the application. Such products can result, for example, from oxidation, reduction, hydrolysis, am idation, deam idation, esterification, deesterification, enzymatic cleavage, and the like, of an administered compound. Accordingly, the present application includes metabolites of compounds of the application, whether prepared by mutation of a compound of the application or prepared by modification of a mammal with a compound of the application for a time and under conditions sufficient to produce the metabolite.

[0086] The present application further includes within its scope prodrugs of the compounds of the application. In general, such prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the desired therapeutically active compound. Thus, in these cases, the term "administering" shall encompass the treatment of the various ailments or conditions described with prodrug forms of one or more of the claimed compounds that readily undergo modifications in vivo to yield the parent compound. Conventional procedures for the selection and

[0087] The present application further includes within its scope stable isotopic derivatives of the compounds of the application, which are identical to the compounds of the application except that one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number usually found in nature.

[0088] The present application also encompasses compounds of the application containing protecting groups. During any process for the preparation of a compound of the application, protecting groups on any reactive groups on any molecule involved in the process can be necessary and / or desirable. This forms a chemically protected form of a compound of the application. This can be achieved by means of conventional protecting groups, for example, those described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which are incorporated herein by reference. The protecting groups can be removed at a suitable subsequent stage using methods known from the art.

[0089] The present application includes pharmaceuticals comprising a therapeutic amount of a compound of the present application, and one or more pharmaceutically acceptable carriers and / or excipients. Carriers include, for example, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof. The carrier or excipient can further include time delay materials known in the art, such as glyceryl monostearate or glyceryl distearate, alone or with a wax, ethyl cellulose, hydroxypropylmethyl cellulose, methy lmethacrylate, and the like. If desired, the composition can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The composition can be a liquid, suspension, emulsion, tablet, pill, capsule, sustained release formulation, or powder. The composition can be formulated into a suppository for rectal administration using a traditional binders and carriers such as triglycerides. Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, and magnesium carbonate, among many others. The formulations can be designed to be mixed, granulated and compressed into tablets or dissolved into capsules. In another approach, the composition can be formulated into nanoparticles.

[0090] The active ingredients of the pharmaceutical compositions according to the application can have systemic and / or local action and can therefore be administered in suitable routes, such as oral, parenteral, pulmonary, nasal, sublingual, lingual, buccal, rectal, transdermal, conjunctival, topical or in the form of implants. The active ingredients can also be administered in the form of administration suitable for these routes of administration. Suitable for oral administration are the well-known administration forms which rapidly and / or in a modified manner release the active ingredients, such as tablets (uncoated tablets or coated tablets, such as tablets with enteric coating or with a mucoadhesive coating), capsules, sugar-coated tablets, granules, pellets, powders, emulsions, suspensions and aerosols. Parenteral administration can avoid the absorption step (intravenous, intraarterial, intracardiac, intraspinal or intrathecal administration) or include absorption (intramuscular, subcutaneous, intradermal, transdermal or intraperitoneal administration). Suitable for parenteral administration are in particular solutions, suspensions, emulsions, lyophilisates and sterile powders in the form of preparations for injection and infusion. Suitable for other routes of administration are, for example, drugs for inhalation (in particular powder inhalation, spray), nasal drops / solutions, sprays; tablets or capsules for lingual, sublingual or buccal administration, suppositories, preparations for the ear and the eye, vaginal capsules, aqueous suspensions (lotions, shake-mixes), lipophilic suspensions, ointments, creams, emulsions, pastes, powders or implants, such as stents. The active ingredients can be converted into the stated administration forms by means of methods known per se. They can be achieved with the aid of inert, nontoxic, suitable pharmaceutical excipients. They include in particular carriers (for example microcrystalline cellulose), solvents (for example liquid polyethylene glycols), emulsifiers (for example sodium dodecyl sulfate), dispersants (for example polyvinylpyrrolidone), synthetic and natural biopolymers (for example proteins), stabilizers (for example antioxidants and ascorbic acid), colorants (for example inorganic pigments such as iron oxides) or flavorings and / or odor correctives. In suitable cases, the active ingredients can be present in the form of microencapsulations in one or more of the above-mentioned carriers. The pharmaceutical preparations described above can comprise, in addition to the compounds of the formula M according to the application, further pharmaceutically active ingredients.

[0091] Preparation method

[0092] The present application also relates to a method for the synthesis of a compound of the formula M according to the application, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt, a polymorph, a co-crystal or a solvate of said compound, or a stable isotopic derivative, a metabolite or a prodrug of said compound, comprising:

[0093] Process scheme I: Synthesis of intermediates 1 to 6

[0094]

[0095] Process scheme II:

[0096] Synthesis of intermediates 7-9, 13-1, 13-2, 14-1, 14-2, 15, 16-1, 16-2, 17-1, 17-2, 18-1, 18-2

[0097]

[0098] Process Scheme III:

[0099] Synthesis of intermediates 19-25

[0100]

[0101] Process Scheme IV:

[0102] Synthesis of intermediates 26-29

[0103] DETAILED DESCRIPTION

[0104] The experimental methods used in the following examples are conventional unless otherwise specified.

[0105] The materials, reagents, etc. used in the following examples are commercially available unless otherwise specified.

[0106] The following examples are preferred illustrative preferred embodiments of the present application and do not constitute any limitation on the present application.

[0107] Example 1 Synthesis of 2-(oct-7-yn-1-yl)isoindoline-1,3-dione (1)

[0108]

[0109] Ethynyllithium ethylenediamine complex (11.2 g, 130 mmol) was added to 100 mL of anhydrous dimethylsulfoxide at room temperature, stirred to dissolve, and protected by nitrogen. The reaction solution was cooled to 0°C, and 1-chloro-6-bromo (11.2 g, 130 mmol) was added dropwise to the reaction solution, which was then stirred at room temperature for 20 hours. After the reaction was completed, 50 mL of saturated aqueous ammonium chloride solution was added dropwise to the reaction solution in an ice bath to quench the reaction. The solution was partitioned with saturated brine and diethyl ether, and the aqueous phase was extracted with diethyl ether, and the organic phases were combined. The organic phase was washed with saturated citric acid, saturated brine, and dried. Concentration yielded a colorless clear liquid. The colorless clear liquid was added to DMF, and potassium phthalimide (20 g, 110 mmol) was added. The reaction solution was then stirred at 80°C for 18 hours. The reaction was monitored by TLC, and the solvent was removed by evaporation under reduced pressure. The residue was dissolved in 60 mL of ethyl acetate, partitioned with 30 mL of water, and the organic phase was collected and dried by evaporation under reduced pressure to obtain a white solid.1 H NMR (400 MHz, DMSO-d6) δ 7.87 - 7.65 (m, 4H), 2.68 (s, 1H), 2.08 (d, J = 6.5 Hz, 2H), 1.52 (d, J = 6.3 Hz, 2H), 1.45 - 1.15 (m, 6H).

[0110] Example 2 Synthesis of methyl 5-(6-((tert-butoxycarbonyl)amino)hexyl)isoxazole-3- carboxylate (2)

[0111]

[0112] Intermediate 1 (11.2 g, 130 mmol) was added to a mixture of ethanol and water at room temperature, stirred to dissolve, hydrazine hydrate (11.2 g, 130 mmol) was added dropwise to the reaction solution, and then the reaction solution was stirred at 75 °C for 2 hours. After the reaction was completed, 12N aqueous hydrochloric acid was added dropwise to the reaction solution at an ice bath to ph = 2. Filtration was performed, and the filtrate was collected. The solvent was removed by evaporation under reduced pressure, and the residue was dissolved in 0.1N aqueous hydrochloric acid solution. The aqueous phase was extracted with DCM, and the aqueous phase was collected. The aqueous phase was adjusted to ph = 14 with NaOH solid, and the aqueous phase was extracted with diethyl ether. The organic phase was collected, and the solvent was removed by evaporation under reduced pressure to obtain a colorless clear liquid. The colorless clear liquid was added to DCM, and then Boc anhydride (20 g, 110 mmol) and TEA were added dropwise in sequence. The reaction solution was stirred at room temperature for 18 hours. After the reaction was completed, the reaction was quenched with saturated aqueous ammonium chloride solution, water was added, and the organic phase was collected by liquid separation. The organic phase was washed with saturated citric acid, saturated brine, and dried. Concentration was performed to obtain a colorless liquid. The colorless liquid was added to a mixed solution of ethyl acetate and water, stirred to dissolve, and then sodium bicarbonate was added. The reaction solution was cooled to 0 °C, and ethyl chloro oxime acetate (11.2 g, 130 mmol) in ethyl acetate was added dropwise. The reaction solution was stirred at room temperature for 10 hours. Ethyl chloro oxime acetate (11.2 g, 130 mmol) in ethyl acetate was added dropwise in batches, and the reaction solution was stirred at room temperature for 8 hours. After the reaction was completed, water was added to the reaction solution, and the organic phase was collected by liquid separation. The organic phase was washed with saturated citric acid, saturated brine, and dried. Concentration was performed to obtain a colorless clear liquid. Column chromatography was performed to obtain a white solid. 1 H NMR (400 MHz, Chloroform-d) δ 4.40 (d, J = 14.5 Hz, 2H), 3.08 (s, 2H), 2.79 (d, J = 7.4 Hz, 2H), 1.42 - 1.40 (m, 11H), 1.37 (d, J = 9.1 Hz, 4H). ESI-MS (m / z): 326.18 [M+H] +

[0113] Synthesis of (S)-2-(5-(6-((tert-butoxycarbonyl)amino)hexyl)isoxazole-3- carboxamido)-3-(4-fluorophenyl)propanoic acid methyl ester (3)

[0114]

[0115] Intermediate 2 (1.83 g, 6 mmol) was added to 10 mL of tetrahydrofuran at 0 °C and stirred to dissolve. Lithium hydroxide (0.4 g, 9.6 mmol) in 10 mL of water was added to the reaction solution in batches. Then the reaction solution was stirred at room temperature for 4 hours. After the reaction was completed, the reaction solution was adjusted to pH 7 with saturated citric acid solution. The solvent was removed under reduced pressure, and the residue was added to 10 mL of ice water, and the aqueous phase was adjusted to pH 2 with saturated citric acid solution. Dichloromethane (10 mL x 2) was extracted, and the organic phase was combined, washed with 0.01 N hydrochloric acid, saturated brine, and dried. Concentration gave a white solid. The white solid was added to 50 mL of anhydrous dichloromethane and stirred to dissolve. The reaction solution was cooled to 0 °C, and 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.70 g, 24 mmol), 1- hydroxybenzotriazole (3.20 g, 24 mmol) were added to the reaction solution. The reaction was carried out at 0 °C for 1 h. L-4-fluorophenylalanine monomethyl ester (4.70 g, 24 mmol) was added to the reaction solution. N,N-diisopropylethylamine (DIPEA) (7.8 g, 60 mmol) was further added dropwise to the reaction solution. Then the reaction solution was stirred at room temperature for 8 hours. After the reaction was completed, 20 mL of saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction. The organic layer was collected by separation, washed with saturated aqueous citric acid solution, saturated aqueous sodium bicarbonate solution, saturated brine, and dried. Concentration gave a white solid 4 crude product. Column chromatography separation gave a white solid (3), 1 H NMR (400 MHz, Chloroform-d) δ 7.22 - 7.15 (m, 1H), 7.12 (s, 2H), 6.96 (s, 2H), 6.39 (s, 1H), 4.99 (s, 1H), 3.72 (s, 3H), 3.10 (s, 4H), 2.78 (s, 2H), 1.69 (s, 2H), 1.43 (s, 11H), 1.35 (s, 4H). ESI-MS (m / z): 491.24 [M+H] +

[0116] Synthesis of 5-(tert-butyl) 1-methyl ((S)-2-(5-(6-((tert-butoxy- carbonyl)amino)hexyl)isoxazole-3-carboxamido)-3-(4-fluorophenyl)propanoyl)- glutamate (4)

[0117]

[0118] Intermediate 4 (1.83 g, 6 mmol) was added to 10 mL of tetrahydrofuran at 0 °C and stirred to dissolve. Lithium hydroxide (0.4 g, 9.6 mmol) in 10 mL of water was added to the reaction solution in batches. Then the reaction solution was stirred at room temperature for 4 hours. After the reaction was completed, the reaction solution was adjusted to pH 7 with saturated citric acid solution. The solvent was removed under reduced pressure, and the residue was added to 10 mL of ice water, and the aqueous phase was adjusted to pH 2 with saturated citric acid solution. Dichloromethane (10 mL x 2) was extracted, and the organic phase was combined, washed with 0.01 N hydrochloric acid, saturated brine, and dried. Concentration gave a white solid. The white solid was added to 50 mL of anhydrous dichloromethane and stirred to dissolve. The reaction solution was cooled to 0 °C, and 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.70 g, 24 mmol), 1- hydroxybenzotriazole (3.20 g, 24 mmol) were added to the reaction solution. The reaction was carried out at 0 °C for 1 h. Tert-butyl-L-glutamic acid methyl ester hydrochloride (4.70 g, 24 mmol) was added to the reaction solution. N, N- diisopropylethylamine (DIPEA) (7.8 g, 60 mmol) was added dropwise to the reaction solution. Then the reaction solution was stirred at room temperature for 8 hours. After the reaction was completed, 20 mL of saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction. The organic layer was collected by separation, washed with saturated aqueous citric acid solution, saturated aqueous sodium bicarbonate solution, saturated brine, and dried. Concentration gave a white solid 4 crude product. 50 ml of petroleum ether was added to the slurry, and the solid was filtered to obtain a white solid (4), 1 H NMR (400 MHz, Chloroform-d) δ 7.33 (s, 1H), 7.18 (s, 2H), 6.96 (s, 2H), 6.80 (s, 1H), 6.70 (s, 1H), 6.38 (s, 1H), 4.80 (s, 1H), 4.50 (s, 2H), 3.68 (s, 3H), 3.08 (s, 4H), 2.75 (s, 2H), 2.14 (d, J = 40.5 Hz, 4H), 1.67 (s, 2H), 1.42 (s, 24H). ESI-MS (m / z): 676.35 [M+H] +

[0119] Example 5 Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-2,5,10-trioxo-3,6,11-triaza- 1(3,5)-isoxazolocyclododecan-7-carboxylic acid methyl ester (5)

[0120]

[0121] Intermediate 5 was added to 8 mL of dichloromethane and stirred to dissolve at room temperature. The reaction was cooled to 0 °C and 8 mL of trifluoroacetic acid was added dropwise. The reaction was stirred at room temperature for 12 h. After the reaction was completed, the reaction was spin-dried and the residue was added to 15 mL of DMF, cooled to 0 °C, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.70 g, 24 mmol), 1-hydroxybenzotriazole (3.20 g, 24 mmol) were added. The reaction was stirred at 0 °C for 1 h and the reaction was added dropwise to a mixture of DCM and DIPEA (600 mL / 10 mL) pre-cooled at -40 °C. The reaction was allowed to warm to room temperature and stirred slowly for 12 h. After the reaction was completed, 20 mL of saturated aqueous ammonium chloride was added to quench the reaction. The organic layer was collected after the layers were separated and the organic layer was washed with saturated aqueous citric acid, saturated aqueous sodium bicarbonate, and the organic layer was collected and spin-dried. The residue was dissolved in 20 mL of EA, washed with water, washed with saturated aqueous sodium chloride, and dried. Concentration gave crude 4 as a white solid. Column chromatography gave a white solid (5) 1 H NMR (400 MHz, DMSO-d6) δ 8.94 (d, J = 12.1 Hz, 2H), 7.87 (s, 1H), 7.34 (s, 2H), 7.06 (s, 2H), 6.67 (s, 1H), 4.44 (s, 1H), 4.20 (s, 1H), 3.65 (s, 3H), 3.28 - 2.67 (m, 6H), 2.13 (s, 2H), 1.87 (d, J = 10.9 Hz, 2H), 1.62 (s, 2H), 1.24 (s, 6H). ESI-MS (m / z): 502.22 [M+H] +

[0122] Example 6 Synthesis of (4S,7S,Z)-7-(2-chloroacetyl)-4-(4-fluorobenzyl)-3,6,11- triaza-1(3,5)-isoxazolacycloheptadecane-2,5,10-trione (6)

[0123]

[0124] To a three-necked flask was added Intermediate 5 (10 g, 35 mmol), THF (200 mL) and chloroiodomethane (10.2 mL, 140 mmol) at room temperature, protected with nitrogen and the solution was cooled to -77 °C. LDA (140 mL, 210 mmol, 1.5 M mono-THF complex in cyclohexane) was added dropwise through a pressure-equalizing dropping funnel at a rate to maintain the internal temperature below -70 °C. After the addition was complete, the reaction was stirred for an additional 1 h and quenched with NaHC03. A mixture of AcOH (33 mL) and THF (200 mL) was added at a rate to moderate the temperature to maintain the internal temperature below -65 °C. After the complete addition, the dark suspension was stirred for 10 min and then warmed to ambient temperature. The reaction was diluted with ethyl acetate (500 mL) and the organics were washed with water (250 mL), saturated. NaHC03(250 mL) and brine (250 mL) were dried over magnesium sulfate, filtered, and the solvent was removed in vacuo to yield sodium sulfate. The crude product, a black oil, was purified by flash chromatography eluting with ethyl acetate. The resulting solid was triturated with diethyl ether to yield the title compound as a light yellow solid.

[0125] ESI-MS (m / z): 520.19 [M+H] +

[0126] Example 7 Synthesis of ethyl 1-(6-(((tert-butoxycarbonyl)amino)hexyl)-1H-1,2,3- triazole-4-carboxylate (7)

[0127]

[0128] Tert-butyl (6-hydroxyhexyl)carbamate (1.83 g, 6 mmol) was added to 60 ml DCM and stirred to dissolve. The reaction was cooled to 0 °C and triethylamine (1.83 g, 6 mmol) was added dropwise. Methylsulfonyl chloride (0.4 g, 9.6 mmol) was added dropwise to the reaction over 1 h. The reaction was then stirred at 0 °C for 2 h. Upon completion of the reaction, the reaction was concentrated and the residue was added to 30 ml DMF and stirred to dissolve. Sodium azide (0.4 g, 9.6 mmol) was added to the reaction and the reaction was stirred at 80 °C for 18 h. Upon completion of the reaction, the solvent was removed by evaporation under reduced pressure and the residue was dissolved in 50 ml DCM. The organic phase was washed with water, saturated aqueous ammonium chloride solution, respectively. The organic phase was spin-dried and dissolved in 50 ml n-hexane. The solution was washed with water, saturated brine, and anhydrous sodium sulfate to remove water. The solvent was removed by evaporation under reduced pressure to obtain a colorless liquid. The colorless liquid was added to a mixture of tert-butanol and water and stirred to dissolve. The reaction was cooled to 0 °C and methyl propiolate (1.83 g, 6 mmol), sodium ascorbate (1.83 g, 6 mmol), and copper sulfate pentahydrate (1.83 g, 6 mmol) were added sequentially. The reaction was stirred at room temperature for 12 h. Upon completion of the reaction, the solvent was removed by evaporation under reduced pressure and the residue was dissolved in 100 ml DCM. The solution was washed with water, saturated sodium bicarbonate solution, and saturated brine. The solvent was removed by evaporation under reduced pressure and the residue was added to 100 ml petroleum ether to obtain a white solid by filtration. 1 H NMR (400 MHz, Chloroform-d) δ 8.07 (s, 1H), 4.38 (s, 2H), 3.92 (s, 3H), 3.04 (s, 2H), 1.91 (s, 2H), 1.40 (s, 11H), 1.31 (s, 4H). ESI-MS (m / z): 340.21 [M+H] +

[0129] Example 8 Synthesis of methyl 1-(7-(((tert-butoxycarbonyl)amino)heptyl)-1H-1,2,3- triazole-4-carboxylate (11)

[0130]

[0131] According to the synthesis method of Intermediate 7. 1 H NMR (600 MHz, Chloroform-d) δ 8.07 (s, 1H), 4.38 (s, 2H), 3.92 (s, 3H), 3.04 (s, 2H), 1.91 (s, 2H), 1.40 (s, 11H), 1.31 (s, 4H). ESI-MS (m / z): 340.21 [M+H] +

[0132] Example 9. Synthesis of (S)-methyl 2-(l-(6-((tert-butoxycarbonyl)amino)hexyl)- lH-l,2,3-triazole-4-carboxamido)-3-(4-fluorophenyl)propanoate (8)

[0133]

[0134] According to the method for synthesizing intermediate 3. 1 H NMR (400 MHz, DMSO-d6) δ 8.78 (s, 1H), 8.55 (s, 1H), 7.29 (s, 2H), 7.07 (s, 2H), 6.77 (s, 1H), 4.69 (s, 1H), 4.37 (s, 2H), 3.64 (s, 3H), 3.16 (s, 2H), 2.86 (s, 2H), 1.80 (s, 2H), 1.36 (s, 11H), 1.23 (s, 4H). ESI-MS (m / z): 491.25 [M+H] +

[0135] Example 10. Synthesis of (S)-methyl 2-(l-(7-((tert-butoxycarbonyl)amino)heptyl)- lH-l,2,3-triazole-4-carboxamido)-3-(4-fluorobenzyl)propanoate (13-1)

[0136]

[0137] According to the method for synthesizing intermediate 3.1H NMR (600 MHz, DMSO-d6) δ 8.74 (d, J = 8.2 Hz, 1H), 8.55 (s, 1H), 7.38 - 7.22 (m, 2H), 7.18 - 6.97 (m, 2H), 6.75 (t, J = 5.8 Hz, 1H), 4.79 - 4.63 (m, 1H), 4.38 (t, J = 7.1 Hz, 2H), 3.64 (s, 2H), 3.20 - 3.07 (m, 2H), 2.87 (q, J = 6.6 Hz, 2H), 1.85 - 1.76 (m, 2H), 1.36 (s, 11H), 1.28 - 1.16 (m, 6H). ESI-MS (m / z): 506.27 [M+H] +

[0138] Example 11. Synthesis of (S)-methyl 2-(l-(7-((tert-butoxycarbonyl)amino)heptyl)- lH-l,2,3-triazole-4-carboxamido)-3-(3,4-difluorobenzyl)propanoate (13-2)

[0139]

[0140] According to the method for synthesizing intermediate 3. 1H NMR (600 MHz, DMSO-d6) δ 8.82 (d, J = 8.3 Hz, 1H), 8.54 (s, 1H), 7.39 - 7.34 (m, 1H), 7.33 - 7.26 (m, 1H), 7.10 (s, 1H), 6.74 (t, J = 5.4 Hz, 1H), 4.77 - 4.71 (m, 1H), 4.37 (t, J = 7.1 Hz, 2H), 3.65 (s, 3H), 3.21 - 3.10 (m, 2H), 2.90 - 2.85 (m, 2H), 1.84 - 1.78 (m, 2H), 1.36 (s, 11H), 1.22 (d, J = 17.8 Hz, 8H). ESI-MS (m / z): 538.28 [M+H] +

[0141] Synthesis of (S)-2-(l-(8-((tert-butoxycarbonyl)amino)octyl)-lH-l,2,3-triazole-4- carboxamido)-3-(3,4-difluorobenzyl)propanoic acid methyl ester (14-2)

[0142]

[0143] According to the method for synthesizing intermediate 3. 1 H NMR (600 MHz, DMSO-d6) δ 8.82 (d, J = 8.3 Hz, 1H), 8.54 (s, 1H), 7.39 - 7.34 (m, 1H), 7.33 - 7.26 (m, 1H), 7.10 (s, 1H), 6.74 (t, J = 5.4 Hz, 1H), 4.77 - 4.71 (m, 1H), 4.37 (t, J = 7.1 Hz, 2H), 3.65 (s, 3H), 3.21 - 3.10 (m, 2H), 2.90 - 2.85 (m, 2H), 1.84 - 1.78 (m, 2H), 1.36 (s, 11H), 1.22 (d, J = 17.8 Hz, 8H). ESI-MS (m / z): 538.28 [M+H] +

[0144] Synthesis of 5-(tert-butyl)-l-methyl((s)-2-(l-(6-(tert-butoxycarbonyl)amino)hexyl)- lH-l,2,3-triazole-4-carboxamido)-3-(4-fluorobenzyl)propanoyl-glutamic acid (9)

[0145]

[0146] According to the method for synthesizing intermediate 4. 1H NMR (400 MHz, DMSO-d6) δ 8.53 (s, 2H), 8.26 (s, 1H), 7.27 (t, J = 8.4 Hz, 2H), 7.03 (s, 2H), 6.75 (s, 1H), 5.72 (s, 0H), 4.72 (d, J = 12.5 Hz, 1H), 4.34 (s, 3H), 3.60 (s, 3H), 3.02 (s, 2H), 2.84 (s, 2H), 2.21 (d, J = 42.3 Hz, 2H), 1.84 (d, J = 63.0 Hz, 4H), 1.34 (s, 20H), 1.18 (s, 4H). ESI-MS (m / z): 577.28 [M+H] +

[0147] Example 14 Synthesis of 5-(tert-butyl) 1-methyl ((S)-2-(1-(7-((tert-butoxycarbonyl)amino)heptyl)-1H-1,2,3-triazole-4-carboxamido)-3-(4-fluorobenzyl)propanoyl)-L-glutamic acid (15-1)

[0148]

[0149] According to the synthetic method of Intermediate 4. 1 H NMR (600 MHz, DMSO-d6) δ 8.53 (d, J = 12.0 Hz, 2H), 8.27 (s, 1H), 7.30 (dd, J = 8.6, 5.7 Hz, 2H), 7.06 (t, J = 8.9 Hz, 2H), 6.74 (s, 1H), 4.76 (td, J = 9.0, 4.7 Hz, 1H), 4.40 - 4.31 (m, 3H), 3.64 (s, 3H), 3.11 - 3.00 (m, 2H), 2.87 (d, J = 6.5 Hz, 2H), 2.29 (td, J = 7.1, 6.5, 2.1 Hz, 2H), 2.03 - 1.94 (m, 1H), 1.81 (dd, J = 9.4, 5.1 Hz, 3H), 1.37 (d, J = 13.2 Hz, 20H), 1.27 - 1.17 (m, 6H). ESI-MS (m / z): 713.34 [M+Na] +

[0150] Example 15 Synthesis of 5-(tert-butyl) 1-methyl ((S)-2-(1-(7-((tert-butoxycarbonyl)amino)heptyl)-1H-1,2,3-triazole-4-carboxamido)-3-(3,4-difluorobenzyl)propanoyl)-L-glutamic acid (15-2)

[0151]

[0152] Following the synthetic procedure of Intermediate 4. 1 H NMR (600 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.52 (d, J = 7.6 Hz, 1H), 8.34 (d, J = 8.6 Hz, 1H), 7.36 - 7.26 (m, 2H), 7.11 (s, 1H), 6.74 (t, J = 5.4 Hz, 1H), 4.77 (q, J = 4.2 Hz, 1H), 4.37 (t, J = 7.0 Hz, 2H), 4.34 (s, 1H), 3.63 (s, 3H), 3.11 - 3.00 (m, 2H), 2.87 (q, J = 6.6 Hz, 2H), 2.69 (s, 1H), 2.29 (t, J = 8.0 Hz, 2H), 1.96 (d, J = 5.6 Hz, 1H), 1.85 - 1.77 (m, 3H), 1.39 - 1.31 (m, 20H), 1.22 (dt, J = 33.3, 7.0 Hz, 6H). ESI-MS (m / z): 709.37 [M+H] +

[0153] Example 16 Synthesis of 5-(tert-butyl) 1-methyl ((S)-2-(1-(8- ((tert-butoxycarbonyl)amino)octyl)-1H-1,2,3-triazole-4-carboxamido)-3-(3,4- difluorobenzyl)propanoyl)-L-glutamic acid (16-2)

[0154]

[0155] Following the synthetic procedure of Intermediate 4. 1 H NMR (600 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.52 (d, J = 7.6 Hz, 1H), 8.34 (d, J = 8.6 Hz, 1H), 7.36 - 7.26 (m, 2H), 7.11 (s, 1H), 6.74 (t, J = 5.4 Hz, 1H), 4.77 (q, J = 4.2 Hz, 1H), 4.37 (t, J = 7.0 Hz, 2H), 4.34 (s, 1H), 3.63 (s, 3H), 3.11 - 3.00 (m, 2H), 2.87 (q, J = 6.6 Hz, 2H), 2.69 (s, 1H), 2.29 (t, J = 8.0 Hz, 2H), 1.96 (d, J = 5.6 Hz, 1H), 1.85 - 1.77 (m, 3H), 1.39 - 1.31 (m, 20H), 1.22 (dt, J = 33.3, 7.0 Hz, 6H). ESI-MS (m / z): 709.37 [M+H] +

[0156] Example 17 Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-2,5,10-trioxo-1 1Synthesis of methyl H-3, 6, 11-triaza- 1 (4, 1)-triazacycloheptadecane-7-carboxylate (10)

[0157]

[0158] According to the method for synthesizing intermediate 5. 1 H NMR (400 MHz, DMSO-d6) δ 8.87 - 8.58 (m, 3H), 8.54 (s, 1H), 7.59 (d, J = 37.5 Hz, 1H), 7.35 - 7.15 (m, 2H), 7.10 - 6.94 (m, 2H), 4.55 - 4.26 (m, 3H), 4.17 - 4.03 (m, 1H), 3.59 (s, 3H), 3.16 - 3.02 (m, 1H), 2.90 - 2.67 (m, 1H), 2.72 - 2.57 (m, 1H), 2.15 - 1.89 (m, 2H), 1.88 - 1.64 (m, 3H), 1.38 - 0.87 (m, 6H). ESI-MS (m / z): 502.23 [M+H] +

[0159] Example 18 Synthesis of (4S, 7S, Z)-4-(4-fluorobenzyl)-2, 5, 10-trioxo-1 1 Synthesis of methyl H-3, 6, 11-triaza- 1 (4, 1)-triazacyclooctadecane-7-carboxylate (17-1)

[0160]

[0161] According to the method for synthesizing intermediate 5. 1H NMR (600 MHz, DMSO-d6) δ 8.76 (s, 1H), 8.60 (d, J = 10.4 Hz, 2H), 7.46 (s, 1H), 7.33 - 7.28 (m, 2H), 7.05 (t, J = 8.9 Hz, 2H), 4.64 (ddd, J = 10.1, 8.5, 4.6 Hz, 1H), 4.47 (ddd, J = 11.6, 7.4, 3.8 Hz, 1H), 4.39 (ddd, J = 13.4, 7.3, 4.1 Hz, 1H), 4.25 (ddd, J = 10.4, 6.9, 3.9 Hz, 1H), 3.62 (s, 3H), 3.17 (dd, J = 14.0, 4.6 Hz, 1H), 3.03 (dd, J = 14.1, 10.2 Hz, 1H), 2.75 (dt, J = 21.1, 8.0 Hz, 2H), 2.16 - 2.08 (m, 2H), 1.94 (ddd, J = 13.0, 6.2, 2.6 Hz, 1H), 1.83 (s, 2H), 1.76 (tt, J = 9.0, 5.7 Hz, 1H), 1.24 (s, 2H), 1.08 (t, J = 7.0 Hz, 4H), 0.96 (s, 2H). ESI-MS (m / z): 517.25 [M+H] +

[0162] Example 19 Synthesis of (4S,7S,Z)-4-(3,4-difluorobenzyl)-2,5,10-trioxo-1 1 Synthesis of methyl H-3,6,11-triaza-1(4,1)-triazacyclooctadecane-7-carboxylate (17-2)

[0163]

[0164] Following the procedure for the synthesis of Intermediate 5. 1H NMR (600 MHz, DMSO-d6) δ 8.76 (d, J = 6.8 Hz, 1H), 8.67 (d, J = 8.6 Hz, 1H), 8.59 (s, 1H), 7.47 (t, J = 5.7 Hz, 1H), 7.37 - 7.31 (m, 1H), 7.27 (d, J = 10.8 Hz, 1H), 7.11 (s, 1H), 4.66 (td, J = 8.9, 4.8 Hz, 1H), 4.47 (td, J = 8.3, 7.3, 3.8 Hz, 1H), 4.42 - 4.36 (m, 1H), 4.24 (dt, J = 6.1, 2.9 Hz, 1H), 3.62 (s, 3H), 3.18 (dd, J = 13.9, 4.7 Hz, 1H), 3.06 - 3.00 (m, 1H), 2.80 - 2.68 (m, 2H), 2.12 (q, J = 5.9 Hz, 2H), 1.98 - 1.90 (m, 1H), 1.83 (s, 2H), 1.79 - 1.71 (m, 1H), 1.13 - 0.91 (m, 8H). ESI-MS (m / z): 535.25 [M+H] +

[0165] Example 20 (4S,7S,Z)-4-(3,4-difluorobenzyl)-2,5,10-trioxo-1 1 Synthesis of methyl H-3,6,11-triaza-1(4,1)-triazacyclononadecane-7-carboxylate (18-2)

[0166]

[0167] Following the procedure for the synthesis of Intermediate 5. 1 H NMR (600 MHz, DMSO-d6) δ 8.65 (s, 1H), 8.41 (s, 1H), 7.40 - 7.29 (m, 2H), 7.21 (s, 1H), 7.13 (s, 1H), 4.63 (ddd, J = 9.1, 5.5, 3.9 Hz, 1H), 4.53 - 4.45 (m, 1H), 4.41 - 4.30 (m, 2H), 3.62 (s, 3H), 3.07 (qd, J = 13.9, 7.5 Hz, 2H), 2.97 (dd, J = 13.3, 6.4 Hz, 1H), 2.83 - 2.76 (m, 1H), 2.15 - 1.97 (m, 3H), 1.89 - 1.78 (m, 2H), 1.77 - 1.68 (m, 1H), 1.24 - 0.91 (m, 10H). ESI-MS (m / z): 549.25 [M+H] +

[0168] Example 21 Synthesis of ethyl 5-(2-(3-((tert-butoxycarbonyl)amino)propoxy)ethyl)isoxazole-3- carboxylate (19)

[0169]

[0170] tert-Butyl (3-hydroxypropyl)carbamate (10 g, 57.1 mmol) was dissolved in dichloromethane at 0 °C, triethylamine (14.44 g, 0.14 mol) was added dropwise slowly, the reaction was stirred at 0 °C for 1 hour, then MSC (8.5 g, 74.19 mmol) was added dropwise slowly, the reaction was stirred for 8 hours. After the reaction was completed, the reaction was extracted with water, saturated citric acid solution and saturated sodium chloride solution in turn, the organic phase was spin-dried to obtain a yellow transparent solid. The yellow solid product (14.46 g, 57.07 mmol) was dissolved in a small amount of toluene, 3-butyn-1-ol (8.0 g, 0.11 mol) was dissolved in toluene, heated to 60 °C, after 15 minutes, tetrabutylammonium bromide (917.2 mg, 2.85 mmol) was added and stirred for 15 minutes. Then sodium hydroxide solid (2.97 g, 74.19 mmol) was added, and the temperature was raised to 75 °C. After 30 minutes, the toluene solution of the yellow solid product was added dropwise, and after the addition was completed, the temperature was raised to 85 °C, and the stirring was continued for 8 hours. After the reaction was completed, the reaction system was washed with saturated ammonium chloride solution, and the organic phase was spin-dried to obtain a yellow liquid. The product yellow liquid (9.640 g, 57.07 mmol) and sodium bicarbonate solid (3.12 g, 37.09 mmol) were dissolved in a mixed solution of ethyl acetate and water (100:1), and the temperature was lowered to 0 °C. (Z)-2-chloro-2-(hydroxyimino)acetic acid ethyl ester (5.62 g, 37.09 mmol) was dissolved in 50 ml of ethyl acetate, and the temperature was controlled at 15-20 °C, and added to the above reaction system. After 2 hours, the remaining sodium bicarbonate solid (3.12 g, 37.02 mmol) and (Z)-2-chloro-2-(hydroxyimino)acetic acid ethyl ester (5.62 g, 37.09 mmol) in ethyl acetate were added to the reaction system, and the temperature was controlled at 15-20 °C and the reaction was stirred. After the reaction was completed, the organic phase was washed with water, saturated citric acid aqueous solution and saturated sodium chloride solution in turn, the organic phase was spin-dried, and after purification by column chromatography, intermediate 19 (9.710 g, 45%) was obtained. 1HNMR (400 MHz, Chloroform-d) δ 6.49 (s, 1H), 4.75 (s, 1H), 4.50 - 4.33 (m, 2H), 3.71 (t, J = 6.3 Hz, 2H), 3.49 (d, J = 12.0 Hz, 2H), 3.16 (q, J = 8.0, 7.0 Hz, 4H), 3.11 - 3.00 (m, 2H), 1.79 - 1.65 (m, 2H), 1.41 - 1.37 (m, 12H), ESI-MS (m / z): 243.13 [M+H] +

[0171] Synthesis of (S)-methyl 2-(5-(2-(4-((tert-butoxycarbonyl)amino)propoxy)ethyl)isoxazole-3- carboxamido)-3-(4-fluorobenzyl)propanoate (20)

[0172]

[0173] Following the synthetic procedure of Intermediate 3.1H NMR (400 MHz, DMSO-d6) δ 9.10 (d, J = 8.1 Hz, 1H), 7.30 (dd, J = 8.8, 5.6 Hz, 2H), 7.08 (t, J = 8.9 Hz, 2H), 6.79 (t, J = 5.5 Hz, 1H), 6.55 (s, 1H), 4.67 (ddd, J = 10.1, 8.1, 5.2 Hz, 1H), 3.64 - 3.67 (m, 5H), 3.39 (t, J = 6.3 Hz, 2H), 3.21 - 3.07 (m, 2H), 3.03 (t, J = 6.2 Hz, 2H), 2.93 (q, J = 6.7 Hz, 2H), 1.57 (p, J = 6.5 Hz, 2H), 1.36 (s, 9H). ESI-MS (m / z): 516.20 [M+Na]+

[0174] Synthesis of (S)-methyl 2-(5-(2-(4-((tert-butoxycarbonyl)amino)propoxy)ethyl)isoxazole-3- carboxamido)-3-(3,4-difluorobenzyl)propanoate (21)

[0175]

[0176] Following the synthetic procedure of Intermediate 3. 1H NMR (400 MHz, Chloroform-d) δ 7.08 (s, 1H), 6.98 (s, 1H), 6.87 (s, 1H), 6.51 (s, 1H), 5.00 (s, 1H), 3.76 (s, 3H), 3.74 (s, 2H), 3.51 (s, 2H), 3.24 - 3.10 (m, 4H), 3.06 (s, 2H), 1.74 (s, 2H), 1.44 (s, 6H), 1.25 (s, 3H). ESI-MS (m / z): 534.20 [M+Na] +

[0177] Example 24 Synthesis of 5-(tert-butyl) 1-methyl ((S)-2-(5-(2-(4-((tert- butyloxycarbonylcarbonyl)amino)propoxy)ethylisoxazole-3-carboxamido)-3-(3,4- difluorobenzyl)propanoyl)-L-glutamate (23)

[0178]

[0179] According to the synthetic method of Intermediate 4. 1 H NMR (400 MHz, Chloroform-d) δ 7.08 (s, 1H), 6.98 (s, 1H), 6.87 (s, 1H), 6.51 (s, 1H), 5.00 (s, 1H), 3.76 (s, 3H), 3.74 (s, 2H), 3.51 (s, 2H), 3.24 - 3.10 (m, 4H), 3.06 (s, 2H), 1.74 (s, 2H), 1.44 (s, 6H), 1.25 (s, 3H). ESI-MS (m / z): 534.20 [M+Na] +

[0180] Example 25 Synthesis of 5-(tert-butyl) 1-methyl ((S)-2-(5-(2-(4-((tert- butyloxycarbonylcarbonyl)amino)propoxy)ethylisoxazole-3-carboxamido)-3-(3,4- difluorobenzyl)propanoyl)-L-glutamate (23)

[0181]

[0182] According to the synthetic method of Intermediate 4. 1H NMR (400 MHz, DMSO-d6) δ 8.70 (s, 1H), 8.54 (s, 1H), 7.34 (s, 2H), 7.13 (s, 1H), 6.77 (s, 1H), 6.56 (s, 1H), 4.75 (s, 1H), 4.35 (dt, J = 8.4, 4.3 Hz, 1H), 3.66 (d, J = 8.4 Hz, 5H), 3.42 (s, 2H), 2.98 (d, J = 45.0 Hz, 6H), 2.29 (s, 2H), 1.97 (s, 1H), 1.84 (s, 1H), 1.61 (s, 2H), 1.40 (s, 9H), 1.38 (s, 9H). ESI-MS (m / z): 697.32 [M+H] +

[0183] Example 26 Synthesis of (4S,7S,Z)-methyl 4-(4-fluorobenzyl)2,5,10-trioxo- 15-oxa-3,6,11-triazacycloheptadecane-7-carboxylate (24)

[0184]

[0185] According to the synthesis method of Intermediate 6. 1 H NMR (400 MHz, Methanol-d4) δ 7.25 (s, 2H), 7.00 - 6.92 (m, 2H), 6.64 (s, 1H), 4.81 (s, 1H), 4.26 (s, 1H), 3.69 (s, 5H), 3.37 (s, 2H), 2.95 (d, J = 58.0 Hz, 6H), 2.23 (s, 2H), 1.99 (s, 2H), 1.50 (s, 2H). ESI-MS (m / z): 505.21 [M+H] +

[0186] Example 27 Synthesis of (4S,7S,Z)-methyl 4-(3,4-difluorobenzyl)2,5,10- trioxo-15-oxa-3,6,11-triazacycloheptadecane-7-carboxylate (25)

[0187]

[0188] According to the synthesis method of Intermediate 6. 1H NMR (600 MHz, DMSO-d6) δ 9.47 (d, J = 5.2 Hz, 1H), 8.91 (d, J = 9.1 Hz, 1H), 7.88 (t, J = 5.7 Hz, 1H), 7.38 - 7.22 (m, 2H), 7.08 (t, J = 6.7 Hz, 1H), 6.68 (s, 1H), 4.67 - 4.62 (m, 1H), 4.14 (dt, J = 9.2, 4.7 Hz, 1H), 3.66 (q, J = 5.4 Hz, 2H), 3.62 (s, 3H), 3.04 - 2.76 (m, 4H), 2.20 (qdd, J = 16.7, 7.5, 3.6 Hz, 2H), 1.93 - 1.79 (m, 2H), 1.43 (dddd, J = 16.0, 13.0, 8.1, 5.5 Hz, 2H), 1.23 (d, J = 6.2 Hz, 2H), 0.85 - 0.73 (m, 2H). ESI-MS (m / z): 523.19 [M+H] +

[0189] Example 28 Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-2,5,10-trioxo-3,6,11-triaza- 1(3,5)-isoxazolocyclododec-7-carbaldehyde (I)

[0190]

[0191] Intermediate 5 (1.83 g, 6 mmol) was added to a mixture of tetrahydrofuran and ethanol at room temperature and stirred to dissolve. The reaction was cooled to 0 °C, and calcium chloride (0.4 g, 9.6 mmol) and sodium borohydride (0.4 g, 9.6 mmol) were added in sequence. The reaction was stirred at 20 °C for 6 hours. After the reaction was completed, the reaction was quenched with saturated ammonium chloride. The solvent was removed under reduced pressure, and the residue was added to 10 ml of water to form a slurry, which was filtered and dried. The white solid was added to a mixture of DCM and DMF, and Dess-Martin oxidant (0.4 g, 9.6 mmol) was added. The reaction was stirred at 20 °C for 10 hours. After the reaction was completed, the reaction was quenched with saturated aqueous sodium bicarbonate solution. The reaction was rotary evaporated, and the residue was added to 10 ml of water to form a slurry, which was filtered and dried. The white solid was obtained as a crude product. Column chromatography was used to isolate the white solid. 1H NMR (400 MHz, DMSO-d6) δ 9.08 (s, 1H), 8.94 (d, J = 12.1 Hz, 2H), 7.87 (s, 1H), 7.34 (s, 2H), 7.06 (s, 2H), 6.67 (s, 1H), 4.44 (s, 1H), 4.20 (s, 1H), 3.65 (s, 3H), 3.28 - 2.67 (m, 6H), 2.13 (s, 2H), 1.87 (d, J = 10.9 Hz, 2H), 1.62 - 1.56 (m, 2H), 1.24 - 1.20 (m, 6H). ESI-MS (m / z): 472.21 [M+H] +

[0192] Example 29 (4S,7S,Z)-4-(4-Fluorobenzyl)-2,5,10-trioxo-1 1 H-3,6,11-triaz-1(4,1)-triazolocyclononadecan-7-carbaldehyde (III)

[0193]

[0194] Compound II was synthesized according to the synthetic method of Compound I, using intermediate 10 as the starting material. 1 H NMR (500 MHz, DMSO-d6) δ 9.38 (d, J = 6.5 Hz, 1H), 8.92 - 8.66 (m, 2H), 8.63 (d, J = 6.3 Hz, 1H), 7.74 - 7.56 (m, 1H), 7.34 (dt, J = 13.6, 6.9 Hz, 3H), 7.08 (q, J = 8.5 Hz, 3H), 4.71 - 4.50 (m, 1H), 4.49 - 4.32 (m, 4H), 4.22 - 3.94 (m, 1H), 3.27 - 3.06 (m, 3H), 2.96 - 2.57 (m, 2H), 2.23 - 1.93 (m, 2H), 1.91 - 1.66 (m, 9H), 1.24 (t, J = 20.7 Hz, 3H), 1.13 - 0.91 (m, 4H). ESI-MS (m / z): 472.22 [M+H] +

[0195] Example 30 (4S,7S,Z)-4-(4-Fluorobenzyl)-2,5,10-trioxo-1 1 H-3,6,11-triaz-1(4,1)-triazolocyclononadecan-7-carbaldehyde (III)

[0196]

[0197] Compound III was synthesized according to the synthetic method of Compound I, using intermediate 18-1 as the starting material. 1H NMR (400 MHz, Methanol-d4) δ 9.30 (d, J = 70.5 Hz, 1H), 8.42 (s, 1H), 8.37 (d, J = 21.3 Hz, 1H), 7.30 (s, 2H), 7.00 (s, 2H), 4.61 - 4.51 (m, 1H), 4.46 - 4.36 (m, 1H), 3.82 (s, 2H), 2.86 (s, 4H), 1.90 (s, 6H), 1.64 (s, 2H), 1.22 (s, 8H). ESI-MS (m / z): 500.25 [M+H] +

[0198] Example 31 (4S,7S,Z)-4-(4-Fluorobenzyl)-2,5,10-trioxo-15-oxa-3,6,11-triaza- 1(3,5)-isoxazolacycloheptadecan-7-formaldehyde (IV)

[0199]

[0200] Compound IV was synthesized according to the synthetic method of Compound I, using intermediate 24 as the starting material. 1 H NMR (500 MHz, DMSO-d6) δ 9.39 (s, 1H), 9.16 - 8.76 (m, 1H), 7.96 (s, 1H), 7.33 (s, 2H), 7.08 (s, 2H), 6.78 (s, 1H), 4.57 (s, 1H), 4.22 (d, J = 242.3 Hz, 1H), 3.66 (s, 2H), 3.20 (s, 3H), 3.00 (s, 3H), 2.83 (s, 2H), 2.02 (d, J = 191.1 Hz, 4H), 1.47 (s, 2H). ESI-MS (m / z): 474.19 [M+H] +

[0201] Example 32 (4S,7S,Z)-4-(4-Fluorobenzyl)-2,5,10-trioxo-15-oxa-3,6,11-triaza- 1(3,5)-isoxazolacyclooctadecan-7-formaldehyde (V)

[0202]

[0203] Compound V was synthesized according to the synthetic method of Compound I, using intermediate 29 as the starting material. 1H NMR (500 MHz, DMSO-d6) δ 9.07 (s, 1H), 7.30 (dd, J = 8.7, 5.6 Hz, 3H), 7.12 (t, J = 8.9 Hz, 2H), 6.46 (s, 1H), 6.38 (d, J = 5.0 Hz, 1H), 4.55 (td, J = 10.3, 5.4 Hz, 1H), 4.30 (s, 1H), 4.07 (dd, J = 8.0, 4.4 Hz, 1H), 3.55 (s, 1H), 3.40 (ddd, J = 9.4, 6.4, 2.9 Hz, 2H), 3.33 - 3.25 (m, 1H), 3.06 (s, 3H), 3.02 - 2.95 (m, 1H), 2.86 (s, 2H), 2.44 (ddd, J = 21.5, 13.2, 5.6 Hz, 2H), 2.29 - 2.15 (m, 2H), 1.98 - 1.81 (m, 2H), 1.68 - 1.42 (m, 1H), 1.38 - 1.29 (m, 1H), 1.60 - 1.50 (m, 1H). ESI-MS (m / z): 488.21 [M+H] +

[0204] Example 33 (4S,7S,Z)-N-(ethylsulfonyl)-4-(4-fluorobenzyl)-2,5,10-trioxo-1 1 H-3,6,11-triaza-l(4,l)-triazolocycloheptadecane-7-carboxamide (VI)

[0205]

[0206] Intermediate 9 (1.83 g, 6 mmol) was added to 10 mL of tetrahydrofuran and stirred to dissolve. The temperature was controlled between 15-20°C, and a solution of lithium hydroxide (0.4 g, 9.6 mmol) in 10 mL of water was added dropwise to the reaction solution within 1 h. Then the reaction solution was stirred for 1 h below 20°C. After the reaction was completed, the reaction solution was adjusted to pH 2 with a saturated citric acid solution. The organic phase was extracted with ethyl acetate (10 mL x 2), and the solvent was removed under reduced pressure. The residue was added to 10 mL of water, and the filter cake was obtained by beating and drying. The white solid was added to a mixed solution of dichloromethane and DMF, and stirred to dissolve. The temperature was lowered to 0°C, and HATU (4.70 g, 24 mmol), N,N-diisopropylethylamine (DIPEA) (7.8 g, 60 mmol) were added to the reaction solution. After stirring for 30 min, ethyl sulfonamide, DMAP, and DBU were added in sequence. The reaction was carried out at room temperature for 12 h. After the reaction was completed, the reaction solution was adjusted to pH 2 with a saturated aqueous citric acid solution, and the solvent was removed under reduced pressure. The residue was added to 10 mL of water, and the filter cake was obtained by beating and drying. The white solid was separated by column chromatography. 1HNMR (400 MHz, Methanol-d4) δ 8.35 (s, 1H), 7.32 (s, 2H), 6.95 (s, 2H), 4.72 (s, 1H), 4.46 (d, J = 28.1 Hz, 2H), 4.20 (s, 1H), 3.12 (d, J = 60.5 Hz, 3H), 2.88 - 2.61 (m, 2H), 1.97 (q, J = 44.8, 38.5 Hz, 8H), 1.22 (d, J = 40.6 Hz, 7H). ESI-MS (m / z): 579.23 [M+H] +

[0207] Example 34 (4S,7S,Z)-N-(cyclopropylsulfonyl)-4-(4-fluorobenzyl)-2,5,10-trioxo- 1 1 Synthesis of H-3,6,11-triaza-l(4,l)-triazacyclononadecan-7-carboxamide (VII)

[0208]

[0209] Compound VII was synthesized according to the synthetic method of compound VI, using intermediate 18-1 as a starting material. 1 H NMR (400 MHz, DMSO-d6) δ 11.92 (s, 1H), 8.64 (d, J = 8.8 Hz, 2H), 8.06 (s, 1H), 7.46 - 7.36 (m, 2H), 7.34 (dd, J = 8.5, 5.7 Hz, 3H), 7.06 (t, J = 8.9 Hz, 2H), 4.73 - 4.61 (m, 2H), 4.54 - 4.35 (m, 2H), 4.43 - 4.32 (m, 3H), 4.22 (s, 1H), 3.16 - 2.79 (m, 5H), 2.34 - 1.56 (m, 4H), 1.22 - 0.86 (m, 3H).

[0210] Example 35 (4S,7S,Z)-N-(ethylsulfonyl)-4-(4-fluorobenzyl)-2,5,10-trioxo-15-oxa-3,6,11- triaza-l(3,5)-isoxazacycloheptadecan-7-carboxamide (VIII)

[0211]

[0212] Compound VIII was synthesized according to the synthetic method of compound VI, using intermediate 24 as a starting material. 1H NMR (400 MHz, DMSO-d6) δ 9.10 (s, 1H), 7.96 (d, J = 7.1 Hz, 1H), 7.45 (t, J = 5.3 Hz, 1H), 7.29 (s, 2H), 7.05 (s, 2H), 6.76 (s, 1H), 4.64 (s, 1H), 4.03 (s, 1H), 3.63-3.40 (m, 4H), 3.29 (s, 3H), 2.96 (s, 4H), 2.68 (s, 1H), 1.94 (td, J = 13.9, 11.5, 7.4 Hz, 2H), 1.81 (t, J = 6.4 Hz, 2H), 1.47 (s, 2H), 1.23 (t, 3H). ESI-MS (m / z): 581.20 [M+H] +

[0213] Example 36 Synthesis of (4S,7S,Z)-7-(2-hydroxyacetyl)-4-(4-fluorobenzyl)-3,6,11- triaza-l(3,5)-isoxazolacycloheptadecane-2,5,10-trione (IX)

[0214]

[0215] Intermediate 6 (1.35 g, 2.67 mmol) was added to DMF (25 mL) at room temperature, followed by benzoyl formic acid (521 mg, 3.47 mmol), and freshly ground CsF (933 mg, 6.14 mmol), and the resulting suspension was placed in a preheated oil bath at 65 °C for 4 hours. The reaction was cooled to ambient temperature, diluted with ethyl acetate (200 mL), and washed with water (3 x 50 mL), brine (50 mL), dried over MgSO4, filtered, and the solvent removed in vacuo. The residue was taken up in methanol (120 mL), K2CO3(38 mg, 0.27 mmol) was added, and the suspension was stirred at ambient temperature for 1 hour. The reaction was neutralized by the addition of 1 M hydrochloric acid, and the solvent removed in vacuo. The crude product was isolated by column chromatography as a white solid. ESI-MS (m / z): 502.22 [M+H] +

[0216] Example 37 Synthesis of (4S,7S,Z)-N-(methylsulfonyl)-4-(3,4-difluorobenzyl)-2,5,10- trioxo-15-oxa-3,6,11-triaza-l(3,5)-isoxazolacycloheptadecane-7-carboxylic acid amine (XI)

[0217]

[0218] Sodium hydride was dissolved in 5 ml of tetrahydrofuran solution at 0°C, and amine methyl sulfonate (78 mg, 0.82 mmol) was added to the reaction solution, which was then stirred at 25°C for 4 hours to obtain a mixed system A. N-methyl morpholine was dissolved in 6 ml of tetrahydrofuran at -20°C, and intermediate 22 (200 mg, 0.41 mmol) was added thereto and stirred for 10 minutes, and isobutyl chloroformate (111.4 mg, 0.82 mmol) was added thereto and stirred for 45 minutes, and then the reaction system was added to the mixed system A at 0°C, and stirred at 25°C for 30 minutes and then stirred for 18 hours. After the reaction was completed, the reaction system was dried, dissolved in dichloromethane, and washed with 1 mol / L citric acid solution and saturated sodium chloride solution, and the crude product was purified by column chromatography to obtain a white solid. 1 H NMR (600 MHz, DMSO-d6) δ 9.07 (s, 1H), 8.21 (s, 1H), 7.93 (s, 1H), 7.35 (ddd, J = 12.1, 7.9, 2.2 Hz, 2H), 7.28 (dd, J = 11.0, 8.4 Hz, 1H), 7.11 (s, 1H), 6.72 (s, 1H), 4.65 (td, J = 9.5, 5.0 Hz, 1H), 4.51 - 4.30 (m, 2H), 4.00 (dt, J = 7.3, 4.7 Hz, 1H), 3.72 - 3.65 (m, 2H), 3.03 - 2.94 (m, 4H), 2.94 - 2.83 (m, 2H), 2.75 (s, 3H), 2.02 - 1.81 (m, 4H), 1.50 - 1.45 (m, 2H). ESI-MS (m / z): 586.21 [M+H] +

[0219] Example 38 Synthesis of (4S,7S,Z)-N-(methylsulfonyl)-4-(4-fluorobenzyl)-2,5,10-trioxo- 15-oxa-3,6,11-triaza-l(3,5)iso oxacycloheptadecane-7-carboxamide (X)

[0220]

[0221] Compound X was synthesized according to the synthesis method of compound VI using intermediate 24 as a raw material. 1H NMR (600 MHz, DMSO-d6) δ 9.05 (s, 1H), 8.22 (s, 1H), 7.83 (s, 1H), 7.37 - 7.23 (m, 3H), 7.05 (t, J = 8.9 Hz, 2H), 6.72 (s, 1H), 4.62 (td, J = 9.3, 5.1 Hz, 1H), 3.99 (ddd, J = 7.5, 5.5, 3.8 Hz, 1H), 3.77 - 3.60 (m, 2H), 3.17 (dd, J = 13.8, 5.2 Hz, 2H), 2.99 (dt, J = 11.9, 4.3 Hz, 2H), 2.96 - 2.92 (m, 1H), 2.86 (dd, J = 13.0, 6.2 Hz, 1H), 2.73 (s, 3H), 1.99 - 1.75 (m, 4H), 1.48 (p, J = 6.5 Hz, 2H), 1.23 (s, 2H). ESI-MS (m / z): 568.19 [M+H] +

[0222] Example 39 Synthesis of (4S,7S,Z)-N-(ethylsulfonyl)-4-(3,4-difluorobenzyl)-2,5,10-trioxo- 15-oxa-3,6,11-triaza-l(3,5)isoazocododecan-7-carboxamide (XI I)

[0223]

[0224] Compound XI I was synthesized according to the procedure for the synthesis of compound VI using intermediate 25 as the starting material. 1 H NMR (600 MHz, DMSO-d6) δ 9.05 (s, 1H), 8.22 (s, 1H), 7.83 (s, 1H), 7.37 - 7.23 (m, 3H), 7.05 (t, J = 8.9 Hz, 2H), 6.72 (s, 1H), 4.62 (td, J = 9.3, 5.1 Hz, 1H), 3.99 (ddd, J = 7.5, 5.5, 3.8 Hz, 1H), 3.77 - 3.60 (m, 2H), 3.17 (dd, J = 13.8, 5.2 Hz, 2H), 2.99 (dt, J = 11.9, 4.3 Hz, 2H), 2.96 - 2.92 (m, 1H), 2.86 (dd, J = 13.0, 6.2 Hz, 1H), 2.73 (s, 3H), 1.99 - 1.75 (m, 4H), 1.48 (p, J = 6.5 Hz, 2H), 1.23 (s, 2H). ESI-MS (m / z): 568.19 [M+H] +

[0225] Example 40. Synthesis of (4S,7S,Z)-N-(cyclopropylsulfonyl)-4-(3,4- difluorobenzyl)-2,5,10-trioxo-15-oxa-3,6,11-triazacyclooctadecan-7-carboxamide (XIII)

[0226]

[0227] Compound XIII was synthesized according to the synthetic method of compound VI, using intermediate 25 as the starting material. 1 H NMR (600 MHz, DMSO-d6) δ 11.93 (s, 1H), 7.90 (d, J = 7.5 Hz, 2H), 7.73 (d, J = 7.5 Hz, 2H), 7.42 (t, J = 7.4 Hz, 2H), 7.34 (td, J = 7.5, 1.1 Hz, 2H), 4.30 (dd, J = 7.1, 5.3 Hz, 2H), 4.23 (t, J = 7.1 Hz, 1H), 4.09 - 4.02 (m, 1H), 3.23 (s, 3H), 2.26 (ddd, J = 9.7, 6.4, 3.5 Hz, 2H), 1.95 - 1.70 (m, 2H), 1.40 (s, 9H). ESI-MS (m / z): 612.18 [M+H] +

[0228] Example 41. Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-N-(methylsulfonyl)- 2,5,10-trioxo-11H-3,6,11-trioxa-1(4,1)-triazacyclononadecan-7-carboxamide (XIV)

[0229]

[0230] Compound XIV was synthesized according to the synthetic method of compound XI, using intermediate 18-1 as the starting material. 1 H NMR (600 MHz, DMSO-d6) δ 8.65 (s, 1H), 7.36 (s, 2H), 7.06 (t, J = 8.8 Hz, 2H), 4.70 - 4.64 (m, 1H), 4.51 - 4.33 (m, 2H), 4.12 (s, 1H), 3.17 (d, J = 4.5 Hz, 1H), 3.06 (s, 2H), 2.99 - 2.89 (m, 2H), 2.62 (s, 1H), 1.89 (d, J = 83.3 Hz, 5H), 1.37 - 0.90 (m, 12H). ESI-MS (m / z): 594.23 [M+H] +

[0231] Example 42. Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-N-(ethylsulfonyl)- 2,5,10-trioxo-11H-3,6,11-trioxa-1(4,1)-triazacyclononadecyl-7-carboxamide (XV)

[0232]

[0233] Compound XV was synthesized according to the procedure for synthesis of Compound XI using intermediate 18-1 as the starting material. 1 H NMR (600 MHz, DMSO-d6) δ 8.61 (s, 1H), 8.54 (s, 1H), 7.36 (d, J = 7.1 Hz, 2H), 7.32 (s, 1H), 7.07 - 7.04 (m, 2H), 4.68 (d, J = 6.3 Hz, 1H), 4.53 - 4.43 (m, 2H), 4.39 - 4.35 (m, 1H), 3.13 - 2.87 (m, 6H), 1.98 - 1.79 (m, 6H), 1.24 - 1.08 (m, 10H), 0.98 (d, J = 12.1 Hz, 3H). ESI-MS (m / z): 608.27 [M+H] +

[0234] Example 43. Synthesis of (4S,7S,Z)-4-(3,4-difluorobenzyl)-N-(methylsulfonyl)- 2,5,10-trioxo-11H-3,6,11-trioxa-1(4,1)-triazacyclononadecyl-7-carboxamide (XVI)

[0235]

[0236] Compound XVI was synthesized according to the procedure for synthesis of Compound XI using intermediate 18-2 as the starting material. 1H NMR (600 MHz, DMSO-d6) δ 11.93 (s, 1H), 8.68 (d, J = 8.2 Hz, 1H), 8.60 (s, 1H), 8.22 (d, J = 7.7 Hz, 1H), 7.42 - 7.32 (m, 2H), 7.28 (dt, J = 10.9, 8.5 Hz, 1H), 7.13 - 7.08 (m, 1H), 4.67 (ddd, J = 9.8, 8.1, 5.2 Hz, 1H), 4.47 (dt, J = 13.7, 5.6 Hz, 1H), 4.38 (dt, J = 13.8, 5.6 Hz, 1H), 4.33 (ddd, J = 10.8, 7.7, 3.6 Hz, 1H), 3.13 - 3.03 (m, 2H), 2.99 (dq, J = 12.7, 6.2 Hz, 1H), 2.85 (dq, J = 12.3, 5.9 Hz, 1H), 2.02 (dd, J = 8.9, 6.7 Hz, 2H), 1.90 (dtd, J = 16.5, 8.4, 7.9, 3.6 Hz, 1H), 1.81 (p, J = 6.5 Hz, 2H), 1.74 (ddt, J = 13.8, 10.0, 7.1 Hz, 1H), 1.20 - 1.08 (m, 5H), 1.07 - 0.93 (m, 5H). ESI-MS (m / z): 612.24 [M+H] +

[0237] Example 44 Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-N-(methylsulfonyl)-2,5,10- trioxo-11H-3,6,11-trioxa-1(4,1)-triazacyclooctadecan-7-carboxamide (XVII)

[0238]

[0239] Compound XVII was synthesized according to the procedure for synthesizing the final product XI, using intermediate 17-1 as the starting material. 1H NMR (600 MHz, DMSO-d6) δ 11.87 (s, 1H), 8.66 (d, J = 8.8 Hz, 2H), 8.56 (s, 1H), 7.61 (t, J = 5.8 Hz, 1H), 7.31 - 7.26 (m, 2H), 7.05 - 7.01 (m, 2H), 4.66 (ddd, J = 10.5, 8.7, 4.3 Hz, 1H), 4.46 (dt, J = 13.6, 5.5 Hz, 1H), 4.39 (dt, J = 13.7, 5.3 Hz, 1H), 4.21 (ddd, J = 8.6, 6.3, 4.0 Hz, 1H), 3.23 (s, 4H), 3.02 (dd, J = 14.0, 10.6 Hz, 1H), 2.76 (q, J = 6.8 Hz, 2H), 2.09 (t, J = 6.9 Hz, 2H), 1.90 - 1.73 (m, 4H), 1.17 - 0.93 (m, 8H). ESI-MS (m / z): 580.24 [M+H] +

[0240] Example 45 Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-N-(ethylsulfonyl)-2,5,10-trioxo- 11H-3,6,11-trioxa-l(4,l)-triazacyclooctadecan-7-carboxamide (XVIII)

[0241]

[0242] The final product XVIII was synthesized according to the synthesis method of the final product XI, using intermediate 17-1 as the starting material. 1 H NMR (600 MHz, DMSO-d6) δ 11.74 (s, 1H), 8.74 (s, 1H), 8.65 (d, J = 8.8 Hz, 1H), 8.55 (s, 1H), 7.64 (s, 1H), 7.32 - 7.26 (m, 2H), 7.02 (t, J = 8.7 Hz, 2H), 4.65 (t, J = 11.7 Hz, 1H), 4.48 - 4.42 (m, 1H), 4.42 - 4.36 (m, 1H), 4.19 (s, 1H), 3.23 (d, J = 14.0 Hz, 1H), 3.05 - 2.98 (m, 1H), 2.80 - 2.68 (m, 2H), 2.10 (s, 2H), 1.90 - 1.75 (m, 4H), 1.23 (t, J = 7.1 Hz, 4H), 1.18 - 0.87 (m, 9H). ESI-MS (m / z): 594.25 [M+H] +

[0243] Example 46. Synthesis of (4S,7S,Z)-4-(4-fluorobenzyl)-N-(cyclopropylsulfonyl)- 2,5,10-trioxo-11H-3,6,11-trioxa-1(4,1)-triazacyclooctadecan-7-carboxamide (XIX)

[0244]

[0245] Compound XIX was synthesized according to the procedure for synthesis of final product XI using intermediate 18-1 as the starting material. 1 H NMR (600 MHz, DMSO-d6) δ 11.80 (s, 1H), 8.66 (d, J = 8.7 Hz, 2H), 8.56 (s, 1H), 7.59 (s, 1H), 7.32 - 7.26 (m, 2H), 7.03 (t, J = 8.9 Hz, 2H), 4.66 (ddd, J = 10.5, 8.7, 4.3 Hz, 1H), 4.48 - 4.41 (m, 1H), 4.42 - 4.35 (m, 1H), 4.23 (s, 1H), 3.22 (d, J = 18.0 Hz, 1H), 3.05 - 3.00 (m, 1H), 2.94 (s, 1H), 2.76 (d, J = 6.0 Hz, 2H), 2.09 (t, J = 6.8 Hz, 2H), 1.84 (d, J = 6.5 Hz, 4H), 1.08 (s, 11H). ESI-MS (m / z): 606.27 [M+H] +

[0246] Example 47. Synthesis of (4S,7S,Z)-4-(3,4-difluorobenzyl)-N-(methylsulfonyl)- 2,5,10-trioxo-11H-3,6,11-trioxa-1(4,1)-triazacyclooctadecan-7-carboxamide (XX)

[0247]

[0248] Compound XX was synthesized according to the procedure for synthesis of final product XI using intermediate 17-2 as the starting material. 1H NMR(600MHz,DMSO-d6)δ11.88(s,1H),8.72(d,J=8.7Hz,1H),8.57(s,1H),7.59 (s,1H),7.39–7.31(m,1H),7.26(q,J=8.6Hz,1H),7.10(s,1H),4.72–4.66(m,1H ),4.49–4.43(m,1H),4.43–4.36(m,1H),4.20(s,1H),3.22(d,J=14.9Hz,3H),3. 06–2.99(m,1H),2.77(s,2H),2.08(s,2H),1.81(d,J=38.3Hz,4H),1.09(s,8H).

[0249] Example 48: In vitro enzyme activity assay of SARS-CoV-2 3CL protease inhibitor

[0250] Detection of targets using fluorescence resonance energy transfer (FRET) technique

[0251] The enzyme activity of the SARS-CoV-2 3CL protease inhibitor was determined using a substrate designed based on the Nsp5 protease recognition site: MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2. The final inhibitor concentrations were 2 μM, 1 μM, 500 nM, 250 nM, 125 nM, 62.5 nM, 31.25 nM, 15.625 nM, 7.8 nM, 3.9 nM, and 1.95 nM, with a negative control included. Enzyme activity was measured using 96-well plates. A 100 μL reaction mixture contained: 50 mM Tris-HCl (pH 7.3), 1 mM EDTA, 150 nM SARS-CoV-2-3CLpro, 20 μM fluorescent substrate, and different concentrations of inhibitor. The reaction was carried out at 30 °C, and fluorescence intensity was detected using a microplate reader. The data were processed using GraphPad Prism 5 software to obtain the IC50 of the inhibitor. 50 The experimental results are shown in Table 1.

[0252] The results showed that compound I-IX had good inhibitory activity against SARS-CoV-2 3CL protease, and its IC50 value was [value missing]. 50 It is 32-275 nM.

[0253] Table 1. Compounds inhibit SARS-CoV-2 3CL protease IC 50

[0254]

[0255] Example 21 In vitro enzyme activity experiment of EV71 3C protease inhibitor

[0256] The enzyme activity of the inhibitor against EV71 3C protease was determined by reverse-phase HPLC method. The polypeptide enzyme activity detection substrate (LEVLFQGPSK) was designed according to the 3C protease recognition site, and the final concentration of the inhibitor was 9uM, 3uM, 1uM, 333nM, 111nM, respectively, and a negative control was set. The enzyme activity was determined by using a 96-well plate, and the 100ul reaction system included 50mM Tris-HCl pH7.5, 4uM EV71 3C protease, 2g / L polypeptide substrate and different concentrations of inhibitors. The reaction was carried out at 30℃, and the fluorescence intensity was detected by a microplate reader. The obtained data was processed by software GraphPad Prism 5 to obtain the IC 50 of the inhibitor. The experimental results are shown in Table 2.

[0257] The results show that the IC 50 of compounds I-IX is 0.45-6.28uM. Among them, the enzyme inhibition activities of compounds III, IV and VIII are about twice that of the positive drug lopinavir, and the enzyme inhibition activity of compound IX is at least three times that of the positive drug lopinavir. The results show that the above compounds have good inhibitory activity on EV71 3C protease.

[0258] Table 2 IC of compounds inhibiting EV71 3C protease 50

[0259]

[0260] Although the specific embodiments of the present application have been described in detail, those skilled in the art will understand that various modifications and substitutions can be made to those details according to all the teachings disclosed, and these changes are within the scope of protection of the present application. The entire scope of the present application is given by the appended claims and any equivalents thereof.

Claims

1. A cyclic peptide compound represented by Formula M or a pharmaceutically acceptable salt of the compound, In formula M, R1is selected from wherein R' is C1-C6alkyl or C3-C6cycloalkyl, R2 is a phenyl ring substituted with 1, 2, 3 or 4 substituents selected from the group consisting of fluorine, chlorine, bromine, iodine; Y is selected from C, O; n+m = 5, 6 or 7; A ring is selected from the following 5-membered heteroaryl groups:

2. The compound according to claim 1 or a pharmaceutically acceptable salt of the compound, wherein R' is C1-C3 alkyl or C3-C6 cycloalkyl, R2 is a phenyl ring substituted with 1, 2, 3 or 4 fluorines.

3. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt of the compound, wherein R' is methyl, ethyl or cyclopropyl, R2 is selected from the group consisting of 4-fluorophenyl, 3-fluorophenyl, 3,4- difluorophenyl.

4. The compound according to claim 1 or a pharmaceutically acceptable salt of the compound, wherein the compound has a structure selected from the group consisting of:

5. A process for the preparation of a compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt of such a compound, wherein, The compound has a structure as shown in Formula 6, and the synthesis of the compound of Formula 6 employs the following reaction scheme: wherein A, R2, m and n are as described in any one of claims 1-4; The compound of Formula 5 is synthesized employing the following reaction scheme: wherein A, R2, m and n are as described in any one of claims 1-4, and reaction conditions a and b are inorganic bases.

6. The method according to claim 5, wherein the inorganic base is selected from the group consisting of sodium hydroxide, sodium carbonate, lithium hydroxide.

7. A process for the preparation of a compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt of said compound, wherein, The compound has a structure as shown in Formula 7, and the synthesis of the compound of Formula 7 employs the following reaction scheme: wherein A, R2, R', m and n are as described in any one of claims 1-4, and reaction condition c is an inorganic base; The compound of Formula 5 is synthesized employing the following reaction scheme: wherein A, R2, m and n are as described in any one of claims 1-4, and reaction conditions a and b are inorganic bases.

8. The method according to claim 7, wherein the inorganic base is selected from the group consisting of sodium hydroxide, sodium carbonate, lithium hydroxide.

9. The method of claim 5 or 7, wherein, The compound of Formula 2 has a structure as shown in Formula 2-1, and the compound of Formula 2-1 is prepared by the following reaction scheme:

10. The method of claim 5 or 7, wherein, The compound of Formula 2 has a structure as shown in Formula 2-2, and the compound of Formula 2-2 is prepared by the following reaction scheme:

11. The method of claim 5 or 7, wherein, The compound of Formula 2 has a structure as shown in Formula 2-3, and the compound of Formula 2-3 is prepared by the following reaction scheme:

12. A pharmaceutical composition comprising at least one compound according to any one of claims 1-4 or a pharmaceutically acceptable salt of the compound; optionally, the pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient, carrier, vehicle or adjuvant.

13. The pharmaceutical composition according to claim 12, further comprising an EV71 antiviral agent.

14. The pharmaceutical composition according to claim 13, wherein the EV71 antiviral agent is an antiviral agent selected from the group consisting of 3D protease inhibitors and VP1 protein inhibitors.

15. Use of a compound according to any one of claims 1-4 or a pharmaceutically acceptable salt of the compound in the preparation of a medicament, wherein the medicament is an antiviral medicament. The virus against which the antiviral drug is directed is selected from the group consisting of enterovirus and coronavirus; the enterovirus is coxsackievirus, poliovirus or enterovirus 71, and the coronavirus is SARS-CoV-2, HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV or MERS-CoV.

16. A compound having the structure of Formula 5: ###0005### Formula 5 wherein A, R2, m and n are as described in any one of claims 1 to 4.

17. Use of a compound having the structure of Formula 5 according to claim 16 as an intermediate for the preparation of a cyclic peptide compound as described in any one of claims 1 to 4. ​

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