Macrocyclic lipopeptide compound containing disulfide bond as well as preparation method and application of macrocyclic lipopeptide compound

Abstract

The invention provides a disulfide bond-containing macrocyclic lipopeptide compound as well as a preparation method and application thereof. Specifically, the invention provides a macrocyclic lipopeptide compound as shown in the specification or a pharmaceutically acceptable salt thereof. The compound is wide in antibacterial spectrum, and has excellent in-vitro antibacterial activity (especially aiming at acinetobacter baumannii and pseudomonas aeruginosa), excellent in-vivo pharmacokinetic property and excellent in-vivo efficacy. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmacology, and relates to the fields of drug synthesis and pharmacology. More specifically, it relates to macrocyclic lipopeptide compounds containing disulfide bonds, their pharmaceutical compositions, their preparation methods, and their use in the preparation of anti-infective drugs. Background Art

[0002] The overuse and abuse of antibiotics have led to the rapid development of bacterial drug resistance, seriously threatening human health and becoming a global public health crisis. In the past decade or so, large pharmaceutical companies have significantly reduced their investment in the research and development of new antibacterial drugs, resulting in a relatively lagging development of new-structure and new-mechanism antibacterial drugs, further exacerbating the intensification and spread of bacterial drug resistance.

[0003] Compared with Gram-positive bacteria, the problem of drug resistance in Gram-negative bacteria is more prominent. In particular, infections caused by multi-drug resistant (MDR) and extensively drug resistant (XDR) Gram-negative bacteria have become the main challenges faced by global health institutions. The World Health Organization (WHO) released the first list of antibiotic-resistant "priority pathogens" in 2017, and three of the highest-level critical categories are all Gram-negative bacteria, such as carbapenem-resistant Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter. Infections caused by these bacteria are extremely difficult to treat. However, in the past 50 years, no new-structure and new-mechanism antibacterial drugs effective against drug-resistant Gram-negative bacteria have been launched on the market. Currently, antibacterial drugs against drug-resistant Gram-negative bacteria in the clinical research stage are almost all improvements or upgrades based on existing drugs, unable to avoid cross-resistance, and having a narrow antibacterial spectrum. These situations have led to a dilemma of no available drugs for drug-resistant Gram-negative bacterial infections worldwide. Therefore, the research and development of broad-spectrum antibacterial drugs with a completely new mechanism of action effective against multi-drug resistant bacteria, especially drug-resistant Gram-negative bacteria, has great scientific and social significance.

[0004] Bacterial signal peptidase is an important membrane-bound protease that cleaves and removes the signal sequence after the translocation of bacterial precursor proteins across the cytoplasmic membrane, which is crucial for the survival of bacteria themselves. Aromamycin belongs to macrocyclic lipopeptide compounds and is a class of inhibitors of bacterial signal peptidase I (SPase I). Its novel chemical structure, unique membrane penetration mode, and brand-new mechanism of action enable these compounds to exhibit strong antibacterial activity against both clinically isolated sensitive and drug-resistant Gram-negative bacteria, and there is no cross-resistance with existing clinical antibacterial drugs, showing great development prospects.

[0005] Companies such as Genentech have reported a series of macrocyclic lipopeptide compounds. The most representative compound, G0775, is currently undergoing preclinical studies. However, G0775 has the problem of a narrow antibacterial spectrum. It shows good activity against Escherichia coli and Klebsiella pneumoniae, but poor activity against Acinetobacter baumannii and Pseudomonas aeruginosa. Moreover, the metabolism of G0775 in animals is not ideal, which to a certain extent limits its possibility as a drug for treating systemic infections.

[0006] Therefore, there is a need in the art for an antibacterial compound with a broader antibacterial spectrum, stronger antibacterial activity in vitro and in vivo, and better metabolic properties in animals. Summary of the Invention

[0007] The present invention provides a class of novel macrocyclic lipopeptide compounds. The compounds of the present invention have a broader antibacterial spectrum, stronger antibacterial activity in vitro and in vivo, better metabolic properties in animals, and are significantly superior to existing similar compounds.

[0008] The object of the present invention is to provide a novel macrocyclic lipopeptide compound with a broad antibacterial spectrum, excellent antibacterial activity in vitro (especially against Acinetobacter baumannii and Pseudomonas aeruginosa), excellent in vivo pharmacokinetic properties, and excellent in vivo efficacy, and its preparation method and use.

[0009] In the first aspect of the present invention, there is provided a macrocyclic lipopeptide compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof:

[0010]

[0011] Wherein,

[0012] R1 is a hydroxyl group, an amino group,

[0013] R2 and R3 are each independently H or a substituted or unsubstituted C1-C6 alkyl group; alternatively, R2 and R3 together with the adjacent carbon atom can form a 3- to 7-membered cycloalkane ring or a 3- to 7-membered heteroalkane ring; wherein, the C1-C6 alkyl group is optionally substituted by one or more of the following groups: C1-C6 alkyl group, C1-C6 alkoxy group, halogen, and amino group;

[0014] n is 0, 1, 2, 3, 4, 5, or 6;

[0015] R4 is H, a substituted or unsubstituted C1-C 10 alkyl group, a substituted or unsubstituted 3- to 8-membered cycloalkyl group, or a substituted or unsubstituted C6-C 10 aryl group, and the C1-C 10 alkyl group, 3- to 8-membered cycloalkyl group, C6-C 10The aryl group is optionally substituted by one or more of the following groups: phenyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 alkoxy, C1-C6 alkylthio, and C1-C6 alkylsilyl;

[0016] X is a substituted or unsubstituted C6-C 10 aryl ring or a substituted or unsubstituted 3-8-membered heteroaryl ring, and the C6-C 10 aryl ring or 3-8-membered heteroaryl ring is optionally substituted by one or more of the following groups: C1-C6 alkyl, C1-C6 alkoxy, halogen, and amino;

[0017] Y is a substituted or unsubstituted C6-C 10 aryl ring or a substituted or unsubstituted 3-8-membered heteroaryl ring, and the C6-C 10 aryl ring or 3-8-membered heteroaryl ring is optionally substituted by one or more of the following groups: C1-C6 alkyl, C1-C6 alkoxy, halogen, and amino.

[0018] In another preferred embodiment, the configuration of the chiral carbon atom in the compound is of the R type or the S type.

[0019] In another preferred embodiment, the macrocyclic lipopeptide compound has a structure as shown in general formula (II):

[0020]

[0021] wherein, R1, R2, R3, R4, X, Y, and n are as described herein.

[0022] In another preferred embodiment, R1 is hydroxy,

[0023] In another preferred embodiment, R1 is

[0024] In another preferred embodiment, R2 and R3 are each independently H or a substituted or unsubstituted C1-C4 alkyl.

[0025] In another preferred embodiment, R2 and R3 are each independently H or a C1-C3 alkyl.

[0026] In another preferred embodiment, R2 and R3 are each independently H or methyl.

[0027] In another preferred embodiment, R2 and R3 are each independently H, R-methyl, or S-methyl.

[0028] In another preferred embodiment, R2 and R3 can be connected to the adjacent carbon atom to form a 3-4-membered cycloalkane ring or a 3-4-membered heteroalkane ring.

[0029] In another preferred embodiment, R2 and R3 and the adjacent carbon atoms thereof may be connected to form cyclopropane or cyclobutane.

[0030] In another preferred embodiment, n is 0, 1, 2 or 3.

[0031] In another preferred embodiment, R4 is a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted 3- to 8-membered cycloalkyl group or a substituted or unsubstituted 6-membered aryl group, and the C1-C8 alkyl group, 3- to 8-membered cycloalkyl group, 6-membered aryl group are optionally substituted by one or more of the following: phenyl, C2-C4 alkenyl, C2-C4 alkynyl, halogen, C1-C4 alkoxy, C1-C4 alkylthio and C1-C4 alkylsilyl.

[0032] In another preferred embodiment, R4 is a C1-C8 alkyl group (preferably a C1-C6 alkyl group), a 3- to 6-membered cycloalkyl group or a 6-membered aryl group, wherein the C1-C8 alkyl group is optionally substituted by halogen.

[0033] In another preferred embodiment, R4 is a C1-C6 alkyl group, preferably a C1-C4 alkyl group, more preferably a tert-butyl group.

[0034] In another preferred embodiment, X is a substituted or unsubstituted 3- to 8-membered nitrogen-containing aromatic heterocycle, and the 3- to 8-membered nitrogen-containing aromatic heterocycle is optionally substituted by one or more of the following groups: C1-C6 alkyl, halogen and amino.

[0035] In another preferred embodiment, X is a substituted or unsubstituted 5- to 6-membered aromatic heterocycle containing 1 to 2 nitrogen atoms, and the aromatic heterocycle is optionally substituted by one or more of the following groups: C1-C3 alkyl and halogen.

[0036] In another preferred embodiment, X is a substituted or unsubstituted pyrimidine or pyridine.

[0037] In another preferred embodiment, X is wherein "*" represents the connection site adjacent to the amide, and Ra and Rb are each independently H, halogen or C1-C4 alkyl.

[0038] In another preferred embodiment, X is wherein "*" represents the connection site adjacent to the amide.

[0039] In another preferred embodiment, Y is a substituted or unsubstituted benzene ring, and the benzene ring is optionally substituted by one or more of the following: C1-C6 alkyl and halogen.

[0040] In another preferred embodiment, Y is wherein "*" represents the connection site adjacent to the disulfide bond, and Rc is H, halogen or C1-C4 alkyl.

[0041] In another preferred embodiment, Y is Among them, "*" represents the linking site close to the disulfide bond.

[0042] In another preferred embodiment, the macrocyclic lipopeptide compound has a structure as shown in the general formula (III):

[0043]

[0044] Wherein, R2, R3, R4, n are as described herein;

[0045] Ra, Rb, Rc are each independently H, halogen or C1-C4 alkyl.

[0046] In another preferred embodiment, R1, R2, R3, R4, X, Y, n are each independently the corresponding groups of the compounds in Examples 1-56.

[0047] In another preferred embodiment, the pharmaceutically acceptable salt of the macrocyclic lipopeptide compound as shown in the general formula (I) refers to the salt formed by the compound and an organic acid or an inorganic acid, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, hydrobromic acid, phosphoric acid, nitric acid, and the organic acid is selected from formic acid, acetic acid, propionic acid, trifluoroacetic acid, citric acid, malic acid, fumaric acid, tartaric acid, methanesulfonic acid, benzenesulfonic acid.

[0048] In another preferred embodiment, the pharmaceutically acceptable salt of the macrocyclic lipopeptide compound as shown in the general formula (I) is trifluoroacetate, hydrochloride or sulfate, preferably tetra-trifluoroacetate, tetra-hydrochloride or di-sulfate.

[0049] In another preferred embodiment, the macrocyclic lipopeptide compound as shown in the general formula (I) is selected from the following group:

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056] In the second aspect of the present invention, there is provided a pharmaceutical composition, comprising: one or more of the macrocyclic lipopeptide compounds as shown in the general formula (I) or their pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier.

[0057] In the third aspect of the present invention, there is provided the use of a macrocyclic lipopeptide compound represented by the general formula (I) as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition for preventing and / or treating infectious diseases.

[0058] In another preferred embodiment, the infectious disease is an infectious disease caused by multi-drug resistant bacteria.

[0059] In another preferred embodiment, the infectious disease is selected from the group consisting of skin, soft tissue or upper respiratory tract infections caused by bacteria, and pneumonia, meningitis, endocarditis or septicemia caused by bacteria.

[0060] In another preferred embodiment, the multi-drug resistant bacteria are Gram-negative bacteria.

[0061] In another preferred embodiment, the multi-drug resistant bacteria are selected from the group consisting of Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, or combinations thereof.

[0062] In the fourth aspect of the present invention, there is provided a method for preventing and / or treating infectious diseases, the method comprising the step of administering a macrocyclic lipopeptide compound represented by the general formula (I) as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0063] In another preferred embodiment, the subject is a human or non-human mammal.

[0064] 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 in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 Shows the colony count results of each experimental group in Example 59. DETAILED DESCRIPTION OF THE INVENTION

[0066] Through extensive and in-depth structure-activity relationship studies, the present inventors synthesized a series of new compounds and conducted in vitro and in vivo antibacterial activity tests and animal in vivo metabolism tests. It was unexpectedly found for the first time that a class of macrocyclic lipopeptide compounds with disulfide bond side chains have much better antibacterial activity and drug metabolism properties in vitro and in vivo than G0775, and can be used as a new treatment means for human or animal bacterial infectious diseases. On this basis, the present inventors completed the present invention.

[0067] TERMS

[0068] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0069] As used herein, the terms "comprising", "including", and "containing" are used interchangeably and include not only closed definitions but also semi-closed and open definitions. In other words, the terms include "consisting of" and "consisting essentially of".

[0070] In the present invention, the halogen is F, Cl, Br, or I.

[0071] In the present invention, the term "C1-C6" means having 1, 2, 3, 4, 5, or 6 carbon atoms, "C1-C8" means having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and so on. "3-8 membered" means having 3-8 ring atoms, and so on.

[0072] In the present invention, the term "alkyl" represents a saturated linear or branched hydrocarbon group. For example, the term "C1-C6 alkyl" refers to a straight-chain or branched alkyl having 1 to 6 carbon atoms, including, without limitation, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and hexyl, etc.; preferably ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, and tert-butyl.

[0073] In the present invention, the term "alkenyl" represents a straight-chain or branched hydrocarbon group containing at least one double bond. For example, the term "C2-C6 alkenyl" refers to a straight-chain or branched alkenyl having 2 to 6 carbon atoms and containing one double bond, including, without limitation, vinyl, propenyl, butenyl, isobutenyl, pentenyl, and hexenyl, etc.

[0074] In the present invention, the term "alkynyl" refers to a straight-chain or branched alkynyl containing one triple bond. For example, the term "C2-C6 alkynyl" refers to a straight-chain or branched alkynyl having 2 to 6 carbon atoms and containing one triple bond, including, without limitation, ethynyl, propynyl, butynyl, isobutynyl, pentynyl, and hexynyl, etc.

[0075] In the present invention, the term "alkoxy" represents an -O-(alkyl) group. For example, the term "C1-C6 alkoxy" refers to a straight-chain or branched alkoxy having 1 to 6 carbon atoms, including, without limitation, methoxy, ethoxy, propoxy, isopropoxy, and butoxy, etc.

[0076] In the present invention, the term "alkylthio" represents an -S-(alkyl) group. For example, the term "C1-C6 alkylthio" refers to a straight-chain or branched alkylthio having 1 to 6 carbon atoms, including, without limitation, -SCH3, -SCH2CH3, -SCH2CH2CH3, -SCH(CH3)CH3, etc.

[0077] In the present invention, the term "alkylsilyl" represents a -SiRR'R" group. For example, the term "C1-C6 alkylsilyl" refers to a straight-chain or branched-chain alkylsilyl having 1 to 6 carbon atoms, including, without limitation, monomethylsilyl, dimethylsilyl, trimethylsilyl, dimethyl-ethylsilyl, triethylsilyl, etc.

[0078] In the present invention, the term "cycloalkyl" represents a non-aromatic cyclic hydrocarbon group, which may be saturated or unsaturated. For example, the term "3-8 membered cycloalkyl" refers to a cyclic alkyl group having 3 to 8 carbon atoms in the ring, including, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl, etc. The term "3-6 membered cycloalkyl" has a similar meaning.

[0079] In the present invention, the term "cycloalkane ring" represents a non-aromatic hydrocarbon ring, which may be saturated or unsaturated. For example, the term "3-8 membered cycloalkane ring" refers to a hydrocarbon ring having 3 to 8 carbon atoms in the ring, including, without limitation, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, and cyclodecane, etc. The term "3-6 membered cycloalkane" has a similar meaning.

[0080] In the present invention, the term "heteroalkane ring" refers to a saturated or partially saturated non-aromatic ring having a heteroatom selected from N, S, and O, which may be in a monocyclic or bicyclic form, such as a bridged ring, a fused ring, or a spiro ring form. The heteroalkane ring is preferably a 3-7 membered heteroalkane ring, more preferably a 4-6 membered heteroalkane ring, and even more preferably a 5-6 membered heteroalkane ring. Examples of the heteroalkane ring include, but are not limited to: oxetane, azetidine, tetrahydro-2H-pyran, piperidine, piperazine, tetrahydrofuran, morpholine, and pyrrolidine, etc.

[0081] In the present invention, the term "aryl" represents a hydrocarbon group containing one or more aromatic rings. For example, the term "C6-C10 aryl" refers to an aromatic ring group having 6 to 10 carbon atoms without heteroatoms in the ring, such as phenyl, naphthyl, etc.

[0082] In the present invention, the term "aromatic ring" includes benzene ring, naphthalene ring, etc.

[0083] The term "heteroaromatic ring" refers to a cyclic aromatic ring having 1-3 heteroatoms selected from the group consisting of N, S, and O, which can be monocyclic or in the form of a fused ring. In the present invention, the heteroaromatic ring is preferably a 3-8 membered heteroaromatic ring, more preferably a 5-8 membered one. Examples of the heteroaromatic ring include, but are not limited to: pyridine, pyridazine, pyrimidine, pyrazine, triazine, pyrrole, pyrazole, imidazole, (1,2,3)-triazole, and (1,2,4)-triazole, tetrazole, furan, thiophene, isoxazole, thiazole, oxazole, carbazole, indole, indazole, benzothiophene, benzofuran, benzimidazole, benzotriazole, benzothiazole, benzothiadiazole, benzoxazole, isomerized quinoline, phthalazine, quinoxaline, quinazoline, cinnoline, or naphthyridine, and tetrazole, etc. The heteroaromatic ring can be fused to an aryl, heterocyclic, or cycloalkyl ring, and the ring connected to the parent structure is the heteroaromatic ring.

[0084] The heteroaromatic ring can be optionally substituted or unsubstituted. When substituted, the substituents are preferably one or more of the following groups, which are independently selected from alkyl, deuterated alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, alkylthio, alkylamino, halogen, amino, nitro, hydroxy, mercapto, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkylthio, oxo, amido, sulfonamido, formyl, formamido, carboxyl, and carboxylate groups, etc. In the present invention, in the substituents, "heteroaromatic ring" and "heteroaryl" have the same meaning.

[0085] In the present invention, "amino" refers to having the structure -N(R)(R'), where R and R' can independently represent hydrogen, alkyl or substituted alkyl, cycloalkyl or substituted cycloalkyl, aryl or substituted aryl, heterocyclic or substituted heterocyclic, as defined above. R and R' can be the same or different in the dialkylamine moiety. Examples of amino are NH2, NHCH3, N(CH3)2.

[0086] Unless otherwise specified, the alkyl, cycloalkane ring, cycloalkyl, heteroalkane ring, aryl, aromatic ring, and heteroaromatic ring described herein are substituted and unsubstituted groups. The possible substituents thereon include, but are not limited to: hydroxy, amino, nitro, nitrile, halogen, C1-C6 alkyl, C2-C 10 alkenyl, C2-C 10 alkynyl, C3-C 20 cycloalkyl, C3-C 20 cycloalkenyl, C3-C 20 heterocycloalkyl, C3-C 20 heterocycloalkenyl, C1-C6 alkoxy, aryl, heteroaryl, heteroaryloxy, C1-C 10 alkylamino, C1-C 20 dialkylamino, arylamino, diarylamino, C1-C 10 alkylsulfamoyl, C1-C 10Alkylimino, mercapto, C1-C 10 Alkylthio, C1-C 10 Alkylsulfonyl, guanidino, ureido, cyano, acyl, acyloxy, carboxyl and carboxylate groups. On the other hand, cycloalkyl, heterocycloalkyl, heterocycloalkenyl, aryl and heteroaryl may also be fused to each other.

[0087] In the present invention, the substitution is monosubstitution or polysubstitution, and the polysubstitution is disubstitution, trisubstitution, tetrasubstitution, or pentasubstitution. The disubstitution means having two substituents, and so on.

[0088] In the present invention, the term "plural" independently refers to 2, 3, 4, 5.

[0089] The term "pharmaceutically acceptable salt" refers to a salt (including inner salts such as zwitterions) that has a similar efficacy to the parent compound and is acceptable biologically or otherwise (e.g., neither toxic nor harmful to the subject). Therefore, the embodiments of the present invention provide pharmaceutically acceptable salts of the compounds of the present invention. The term "salt" as used herein refers to any acid salt formed by inorganic and / or organic acids, as well as basic salts formed by inorganic and / or organic bases. The salts of the compounds of the present invention can be formed by methods known to those of ordinary skill in the art, for example, by reacting the compounds of the present invention with a certain amount of acid or base (e.g., an equivalent amount of acid or base) in a medium (e.g., a medium in which the salt can precipitate; or using water as the medium and then freeze-drying).

[0090] Active ingredient

[0091] As used herein, the term "compound of the present invention" or "active ingredient of the present invention" is used interchangeably and refers to a macrocyclic lipopeptide compound represented by the general formula (I), or a pharmaceutically acceptable salt thereof. The term also includes racemates and optical isomers.

[0092] The inventors unexpectedly found that after adding a disulfide bond in the lipophilic region, compared with the prior art without introducing a disulfide bond (such as compound G0775), the in vitro activity, metabolism, and in vivo pharmacodynamic properties have been significantly improved. First, in the in vitro activity test experiment, the compounds of the present invention have comparable activity to the prior art against Escherichia coli and Klebsiella pneumoniae, while the antibacterial activity against Acinetobacter baumannii and Pseudomonas aeruginosa has been increased by 2-16 times. Second, due to the presence of the disulfide bond, the in vivo pharmacokinetic properties of the present invention have been verified, and it is found that the pharmacokinetics is better than that of the prior art without introducing a disulfide bond. Finally, the present invention has also verified that the compounds of the present invention have significantly excellent in vivo pharmacodynamic properties.

[0093] A large number of studies on the structure-activity relationship of aromamycin have been carried out in the prior art (Nature. 2018; 561(7722): 189-194; ACS Med Chem Lett. 2023; 14(11): 1524-1530). It has been found that introducing a basic side chain on its macrocyclic parent nucleus can promote the penetration of the compound through the bacterial outer membrane, and introducing a cyano group at the carbon terminus can serve as a covalent warhead to covalently bind to the lysine residue K146 of bacterial signal peptidase I, further improving the antibacterial activity of this type of compound. The modification of the lipophilic region also has a great impact on the activity.

[0094] On this basis, the inventors made a mechanistic conjecture on the excellent technical effects of the compounds of the present invention: The target bacterial signal peptidase I of the compounds of the present invention is on the inner membrane of Gram-negative bacteria. The compounds of the present invention first utilize the characteristic of the relatively high permeability of the basic side chain to pass through the outer membrane, and then the lipophilic disulfide bond may help the compound to be anchored on the inner membrane first, enabling the whole molecule to interact with the target on the inner membrane.

[0095] In addition, the compounds of the present invention are not conventional compounds such as quinolones or carbapenems. Their targets and mechanisms of action are completely different from those of the marketed drugs such as quinolones or carbapenems. Therefore, they also have a good therapeutic effect on infections caused by drug-resistant bacteria caused by quinolones or carbapenems and other antibacterial compounds.

[0096] Compounds are understood to include their salts. The term "salt" as used herein refers to acid salts or basic salts formed with inorganic or organic acids and bases. In addition, when the compounds in the present invention contain a basic moiety, it includes but is not limited to amino or pyrimidine, and when it contains an acidic moiety, it includes but is not limited to carboxylic acid. The zwitterions ("inner salts") that may be formed are included within the scope of the term "salt". Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, although other salts are also useful, for example, they can be used in the separation or purification steps during the preparation process. The compounds of the present invention may form salts. For example, Compound I reacts with a certain amount such as an equivalent amount of acid or base, and the salt precipitates out in a medium or is obtained by freeze-drying in an aqueous solution.

[0097] The basic fragments contained in the compounds of the present invention, including but not limited to amines, pyridine or imidazole rings, may form salts with organic or inorganic acids. Typical acids that can form salts include acetate, trifluoroacetate, adipate, alginate, ascorbate, aspartate, benzoate, benzenesulfonate, bisulfate, borate, butyrate, citrate, camphorsulfonate, hydrochloride, hydrobromide, hydroiodide, hydroxyethanesulfonate, lactate, maleate, methanesulfonate, naphthalenesulfonate, nicotinate, nitrate, oxalate, pectinate, persulfate, phenylpropionate, phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, sulfonate, tartrate, thiocyanate, toluenesulfonate such as p-toluenesulfonate, etc.

[0098] Certain compounds of the present invention may contain acidic fragments, including but not limited to carboxylic acids, which may form salts with various organic or inorganic bases. Typical bases for forming salts include ammonium salts, alkali metal salts such as sodium, lithium, potassium salts, alkaline earth metal salts such as calcium, magnesium salts and salts formed with organic bases (such as organic amines), as well as salts formed with amino acids such as arginine, lysine, etc. The basic nitrogen-containing groups can form salts with quaternary ammonium halides, such as small molecule alkyl halides (such as chlorides, bromides and iodides of methyl, ethyl, propyl and butyl), dialkyl sulfates (such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and dipentyl sulfate), long-chain halides (such as chlorides, bromides and iodides of decyl, dodecyl, tetradecyl and hexadecyl), aralkyl halides (such as benzyl and phenyl bromides), etc.

[0099] The compounds or salts in the present invention may exist in tautomeric forms (such as amides and imino ethers). All these tautomers are part of the present invention.

[0100] The compounds represented by the general formula (I) of the present invention contain at least 4 chiral centers, and there are enantiomers and diastereomers. For enantiomers, two enantiomers can be obtained by general chiral resolution methods or asymmetric synthesis methods. For diastereomers, they can be separated by methods such as fractional recrystallization or chromatographic separation. The compounds represented by the general formula (I) of the present invention include any one of the above isomers or mixtures thereof.

[0101] The compounds in the present invention, obtained by preparation, separation and purification in sequence, have a weight content equal to or greater than 90%, for example, equal to or greater than 95%, equal to or greater than 99% ("very pure" compounds), which are listed in the text description. Such "very pure" compounds of the present invention are also part of the present invention.

[0102] The preparation method of the compounds represented by the general formula (I) of the present invention

[0103] The preparation methods of the compounds shown in formula (I) of the present invention are specifically described below, but these specific methods do not impose any limitations on the present invention.

[0104] The compounds shown in formula (I) of the present invention can be prepared by the following methods. However, the conditions of this method, such as reactants, solvents, bases, acids, the amounts of compounds used, reaction temperature, reaction time required, etc., are not limited to the following explanations. The compounds of the present invention can also be conveniently prepared by optionally combining various synthesis 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 pertains.

[0105] Route 1

[0106] One of the preferred preparation routes of the compounds of the present invention is as follows, and this route can prepare Compounds 1-8 and 51:

[0107]

[0108] R4 is defined as before.

[0109] a. Starting material I-1 reacts with pivalic anhydride in the presence of a base in a polar aprotic solvent at 0 °C for 10 min to form intermediate I-2. The base can be potassium carbonate, sodium carbonate, triethylamine, N,N-diisopropylethylamine, and the polar aprotic solvent can be 1,4-dioxane, tetrahydrofuran, dimethylformamide (DMF), dimethylacetamide (DMA), acetonitrile. b. Intermediate I-2 reacts with starting material I-3 in the presence of dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium and sodium carbonate under inert gas protection in a mixed solvent of a polar aprotic solvent and water at 90 °C for 1 h to form intermediate I-4. The polar aprotic solvent can be 1,4-dioxane.

[0110] c. Intermediate I-4 reacts in the presence of lithium hydroxide in a mixed solvent of a polar solvent and water at room temperature for 2 h to form intermediate I-5. The polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, ethanol.

[0111] d. Starting material I-6 reacts with methanesulfonyl chloride in the presence of an organic base in a polar aprotic solvent at room temperature for 3 h to form intermediate I-7. The polar aprotic solvent can be 1,4-dioxane, tetrahydrofuran, dimethylformamide (DMF), dimethylacetamide (DMA), acetonitrile.

[0112] e. Intermediate I-5 and Intermediate I-7 react in a polar solvent at room temperature for 10 min in the presence of an organic base to form Intermediate I-8. The organic base can be triethylamine or N,N-diisopropylethylamine, and the polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, ethanol, dimethylformamide (DMF), dimethylacetamide (DMA), or acetonitrile.

[0113] f. Intermediate I-9 (prepared according to CN111386283) reacts in a mixed solvent of a polar solvent and water at room temperature for 1 h in the presence of lithium hydroxide to form Intermediate I-10. The polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, or ethanol.

[0114] g. Intermediate I-10 and aminoacetonitrile hydrochloride react in a polar aprotic solvent at 0 °C for 1 h in the presence of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate and N,N-diisopropylethylamine to form Intermediate I-11. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), acetonitrile, dichloromethane, or a mixed solvent.

[0115] h. Intermediate I-11 reacts in a polar protic solvent at room temperature for 2 h in the presence of palladium on carbon and hydrogen to form Intermediate I-12. The polar protic solvent can be methanol or ethanol.

[0116] i. Intermediate I-8 and Intermediate I-12 react in a polar aprotic solvent at 0 °C for 10 min in the presence of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate and N,N-diisopropylethylamine to form Intermediate I-13. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), or acetonitrile.

[0117] j. Intermediate I-13 reacts in a polar solvent at 35 °C for 2 h in the presence of trifluoroacetic acid to form Compound 1-8. The polar solvent can be hexafluoroisopropanol.

[0118] k. Compound 6 reacts in a polar solvent at room temperature for 30 min in the presence of sodium bicarbonate to form Compound 51. The polar solvent can be water.

[0119] Route 2

[0120] One of the preferred preparation routes of the compounds of the present invention is as follows. This route can be used to prepare Compounds 9-15, 17-20, 23, 30-40, and 52-56:

[0121]

[0122] R2, R3, R4 and n are defined as before.

[0123] a. The starting material and bis(pinacolato)diboron react in the presence of dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium and potassium acetate under an inert gas atmosphere in a polar solvent at 90 °C for 5 h to form intermediate I-14. The polar solvent can be 1,4-dioxane, dimethylformamide (DMF), or dimethyl sulfoxide (DMSO).

[0124] b. Intermediate I-14 and I-3 react in the presence of dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium and sodium carbonate under an inert gas atmosphere in a mixed solvent of a polar aprotic solvent and water at 90 °C for 1 h to form intermediate I-15. The polar aprotic solvent can be 1,4-dioxane.

[0125] c. Intermediate I-15 and thioacetic acid react in the presence of triphenylphosphine and diisopropyl azodicarboxylate under an inert gas atmosphere in a polar aprotic solvent at 0 °C for 30 min to form intermediate I-16. The polar aprotic solvent can be 1,4-dioxane, tetrahydrofuran, dimethylformamide (DMF), or acetonitrile; or intermediate I-15 and thioacetic acid react in the presence of zinc iodide under an inert gas atmosphere in a polar aprotic solvent at room temperature for 24 h to form intermediate I-16. The polar aprotic solvent can be dichloromethane and chloroform.

[0126] d. Intermediate I-16 reacts in the presence of sodium hydroxide in a mixed solvent of a polar solvent and water at 50 °C for 1 h to form intermediate I-17. The polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, or ethanol.

[0127] e. Intermediate I-17 and intermediate I-7 react in the presence of an organic base in a polar solvent at room temperature for 10 min to form intermediate I-18. The organic base can be triethylamine or N,N-diisopropylethylamine. The polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, ethanol, dimethylformamide (DMF), dimethylacetamide (DMA), or acetonitrile.

[0128] f. Intermediate I-18 and intermediate I-12 react at 0 °C for 10 min in a polar aprotic solvent in the presence of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate and N,N-diisopropylethylamine to form intermediate I-19. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), or acetonitrile.

[0129] g. Intermediate I-19 reacts at 35 °C for 2 h in a polar solvent in the presence of trifluoroacetic acid or HCl to form compound 9-15, 17-20, 23, 30-40, or 55. The polar solvent can be hexafluoroisopropanol.

[0130] h. Compound 11, 13, or 15 reacts at room temperature for 30 min in a polar solvent in the presence of sodium bicarbonate to form compound 52, 53, or 54. The polar solvent can be water.

[0131] i. Compound 54 reacts at room temperature for 30 min in a polar solvent in the presence of sulfuric acid to form compound 56. The polar solvent can be water or methanol.

[0132] Route 3

[0133] One of the preferred preparation routes of the compounds of the present invention is as follows, and this route can prepare compounds 16, 21, 22, and 24-29:

[0134] R2, R3, R4, and n are defined as before;

[0135] LG is a leaving group, preferably halogen or p-toluenesulfonate (OTs).

[0136] a. Starting materials I-20 and I-21 react at room temperature for 2 h in a polar aprotic solvent to form intermediate I-22. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), or acetonitrile.

[0137] b. Intermediate I-22 and I-17 (synthesized according to Route 2) react at room temperature for 10 min in a polar solvent in the presence of an organic base to form intermediate I-18. The organic base can be triethylamine or N,N-diisopropylethylamine, and the polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, ethanol, dimethylformamide (DMF), dimethylacetamide (DMA), or acetonitrile.

[0138] c. Intermediate I-18 and intermediate I-12 react at 0 °C for 10 min in a polar aprotic solvent in the presence of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate and N,N-diisopropylethylamine to form intermediate I-19. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), or acetonitrile.

[0139] d. Intermediate I-19 reacts at 35 °C for 2 h in a polar solvent in the presence of trifluoroacetic acid to form compound 16, 21, 22, or 24-29. The polar solvent can be hexafluoroisopropanol.

[0140] Route 4

[0141] One of the preferred preparation routes of the compounds of the present invention is as follows, and this route can prepare compounds 41-46:

[0142]

[0143] X and Y are defined as before;

[0144] R5 is a C1-C3 alkyl group.

[0145] a. Starting materials I-23 and I-24 react under the protection of an inert gas at 90 °C for 1 h in a mixed solvent of a polar aprotic solvent and water in the presence of dichloro[1,1'-bis(diphenylphosphino)ferrocene]palladium and sodium carbonate to form intermediate I-25. The polar aprotic solvent can be 1,4-dioxane.

[0146] b. Intermediate I-25 and thioacetic acid react under the protection of an inert gas at 0 °C for 30 min in a polar aprotic solvent in the presence of triphenylphosphine and diisopropyl azodicarboxylate to form intermediate I-26. The polar aprotic solvent can be 1,4-dioxane, tetrahydrofuran, dimethylformamide (DMF), or acetonitrile.

[0147] c. Intermediate I-26 reacts at 50 °C for 1 h in a mixed solvent of a polar solvent and water in the presence of sodium hydroxide to form intermediate I-27. The polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, or ethanol.

[0148] d. Intermediate I-27 and I-7-6 react at room temperature for 10 min in a polar solvent in the presence of an organic base to form intermediate I-28. The organic base can be triethylamine or N,N-diisopropylethylamine, and the polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, ethanol, dimethylformamide (DMF), dimethylacetamide (DMA), or acetonitrile.

[0149] e. Intermediate I-28 and Intermediate I-12 react at 0 °C for 10 min in a polar aprotic solvent in the presence of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate and N,N-diisopropylethylamine to form Intermediate I-29. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), or acetonitrile.

[0150] f. Intermediate I-29 reacts at 35 °C for 2 h in a polar solvent in the presence of trifluoroacetic acid to form Compound 41-46. The polar solvent can be hexafluoroisopropanol.

[0151] Route 5

[0152] One of the preferred preparation routes of the compounds of the present invention is as follows, and this route can prepare Compounds 47-49:

[0153]

[0154] R2, R3, R4, and n are defined as before.

[0155] a. Intermediate I-18 (prepared according to Route 2) and Intermediate I-30 (prepared according to CN111386283) react at 0 °C for 10 min in a polar aprotic solvent in the presence of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate and N,N-diisopropylethylamine to form Intermediate I-31. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), or acetonitrile.

[0156] b. Intermediate I-31 reacts at room temperature for 1 h in a mixed solvent of a polar solvent and water in the presence of lithium hydroxide to form Intermediate I-32. The polar solvent can be 1,4-dioxane, tetrahydrofuran, methanol, or ethanol.

[0157] c. Intermediate I-32 reacts at 35 °C for 2 h in a polar solvent in the presence of trifluoroacetic acid to form Compounds 47-49. The polar solvent can be hexafluoroisopropanol.

[0158] Route 6

[0159] One of the preferred preparation routes of the compounds of the present invention is as follows, and this route can prepare Compound 50:

[0160]

[0161] R2, R3, R4, and n are defined as before.

[0162] a. Intermediate I-32 (prepared according to Route 5) and aminoacetaldehyde dimethyl acetal react in a polar aprotic solvent at 0 °C for 1 h in the presence of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate and N,N-diisopropylethylamine to form Intermediate I-33. The polar aprotic solvent can be dimethylformamide (DMF), dimethylacetamide (DMA), dimethyl sulfoxide (DMSO), acetonitrile, dichloromethane, or a mixed solvent.

[0163] b. Intermediate I-33 reacts in a polar solvent at 0 °C for 1 h in the presence of hydrochloric acid to form Compound 50. The polar solvent can be hexafluoroisopropanol.

[0164] Pharmaceutical Composition and Administration Method

[0165] The present invention provides a pharmaceutical composition containing one or more of the compounds represented by the general formula (I) as the main active ingredient.

[0166] Due to the excellent broad-spectrum antibacterial activity of the compounds of the present invention, the compounds of the present invention, or their stereoisomers or optical isomers, pharmaceutically acceptable salts, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used for preventing and / or treating (stabilizing, alleviating, or curing) infectious diseases, especially infectious diseases caused by multi-drug resistant bacteria.

[0167] The pharmaceutical composition of the present invention contains the compound of the present invention within a safe and effective amount range and a pharmaceutically acceptable excipient or carrier. The "safe and effective amount" herein refers to: an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Generally, the pharmaceutical composition contains 1 - 2000 mg of the compound of the present invention per dose, more preferably, contains 10 - 200 mg of the compound of the present invention per dose. Preferably, the "per dose" is a capsule or a tablet.

[0168] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" herein means that the components in the composition can be mixed with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers are cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, 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 dodecyl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0169] There is no particular limitation on the administration route of the compounds or pharmaceutical compositions of the present invention. Representative administration routes include (but are not limited to): oral administration, parenteral (intravenous, intramuscular or subcutaneous).

[0170] Solid dosage forms for oral administration include capsules, tablets, pills, powders, granules, and syrups. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or calcium phosphate, or is mixed with the following components: (a) fillers or bulking agents, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, such as hydroxypropylmethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, such as paraffin wax; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0171] Solid dosage forms such as tablets, pills, capsules, pills, and granules can be prepared with coatings and shell materials, such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed and released in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and wax substances. If necessary, the active compound can also be in the form of microcapsules with one or more of the above excipients.

[0172] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers, and emulsifying agents, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0173] In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, and fragrances.

[0174] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecanol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide, agar, or a mixture of these substances.

[0175] The composition for parenteral injection may contain a physiologically acceptable sterile aqueous or non-aqueous solution, dispersion, suspension or emulsion, and a sterile powder for reconstituting into a sterile injectable solution or dispersion. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0176] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds (such as other antibacterial drugs).

[0177] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal in need of treatment (such as a human), wherein the dosage during administration is a pharmaceutically effective dosage. For a person weighing 60 kg, the daily dosage is usually 1 - 2000 mg, preferably 20 - 500 mg. Of course, the specific dosage should also consider factors such as the route of administration and the health status of the patient, which are within the scope of the skills of a skilled physician.

[0178] The main advantages of the present invention include:

[0179] The compounds of the present invention have a broad antibacterial spectrum, excellent in vitro antibacterial activity (especially against Acinetobacter baumannii and Pseudomonas aeruginosa), good in vivo pharmacokinetic properties, and excellent in vivo efficacy.

[0180] Examples

[0181] The present invention will be further illustrated below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions in the following examples are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0182] In all examples, 1 1H-NMR and 13 13C-NMR were recorded on a BRUKER 400 MHz, 500 MHz or 600 MHz nuclear magnetic resonance spectrometer, and the chemical shifts were expressed in δ (ppm); silica gel was used for separation, which was 200 - 300 mesh if not specified, and the ratio of the eluent was by volume. All reagents were of analytical grade if not otherwise specified.

[0183] Preparation Examples

[0184] Example 1: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(methyldithio)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 1)

[0185]

[0186] (a) (4-(Pivaloylthio)phenyl)boronic acid (I-2)

[0187] Under ice bath conditions, 4-mercaptobenzeneboronic acid (I-1) (1.00 g, 6.49 mmol) was added to 10 ml of N,N-dimethylformamide, stirred to dissolve it, and potassium carbonate (2.69 g, 19.48 mmol) and pivalic anhydride (1.33 g, 7.14 mmol) were added successively. The reaction was carried out at ice bath temperature for 10 min. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography [petroleum ether:ethyl acetate = 2:1] to obtain 954 mg of a white solid with a yield of 62%. 1 1H NMR (600 MHz, Chloroform-d) δ 8.23 (d, J = 7.6 Hz, 2H), 7.54 (d, J = 7.7 Hz, 2H), 1.35 (s, 9H).

[0188] (b) Ethyl 4-methyl-2-(4-(pivaloylthio)phenyl)pyrimidine-5-carboxylate (I-4)

[0189] At room temperature, I-2 (916 mg, 3.85 mmol) and ethyl 4-methyl-2-chloropyrimidine-5-carboxylate I-3 (700 mg, 3.50 mmol) were added to 50 ml of 1,4-dioxane, stirred to dissolve them, and 15 ml of an aqueous solution of sodium carbonate (742 mg, 7.00 mmol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (256 mg, 0.35 mmol) were added successively. Under argon protection, the temperature was raised to 90 °C and the reaction was carried out for 1 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography [petroleum ether:ethyl acetate = 25:1] to obtain 478 mg of a white solid with a yield of 38%. 11H NMR (600 MHz, Chloroform-d) δ 9.23 (s, 1H), 8.57 (d, J = 8.4 Hz, 2H), 7.55 (d, J = 8.4 Hz, 2H), 4.45 (q, J = 7.1 Hz, 2H), 2.92 (s, 3H), 1.46 (t, J = 7.1 Hz, 3H), 1.37 (s, 9H).

[0190] (c) 2-(4-Mercaptophenyl)-4-methylpyrimidine-5-carboxylic acid (I-5)

[0191] At room temperature, I-4 (478 mg, 1.34 mmol) was added to 5 ml of tetrahydrofuran and stirred to dissolve it. Then 1 ml of aqueous lithium hydroxide (224 mg, 5.34 mmol) solution was added, and the reaction was carried out at room temperature for 2 h. The reaction was monitored by TLC until completion. Water was added, and the pH was adjusted to 3 using 1 M aqueous HCl. Ethyl acetate was added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 326 mg of a white solid with a yield of 99%. 1 1H NMR (600 MHz, DMSO-d6) δ 9.13 (s, 1H), 8.43 (d, J = 8.3 Hz, 2H), 7.72 (d, J = 8.3 Hz, 2H), 2.78 (s, 3H).

[0192] (d) 4-Methyl-2-(4-(methyldithio)phenyl)pyrimidine-5-carboxylic acid (I-8-1)

[0193] At room temperature, I-5 (30 mg, 0.12 mmol) and triethylamine (31 mg, 0.31 mmol) were added to 2 ml of methanol and stirred to dissolve them. Then methyl methanethiosulfonate I-7-1 (19 mg, 0.15 mmol) was added, and the reaction was carried out at room temperature for 30 min. The reaction was monitored by TLC until completion. Water was added, and the pH was adjusted to 3 using 1 M aqueous HCl. Ethyl acetate was added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure and purified by column chromatography [dichloromethane:methanol = 10:1] to obtain 29 mg of a white solid with a yield of 83%. 1 1H NMR (600 MHz, DMSO-d6) δ 8.96 (s, 1H), 8.42 (d, J = 8.3 Hz, 2H), 7.67 (d, J = 8.2 Hz, 2H), 2.76 (s, 3H), 2.53 (s, 3H).

[0194] (e) (4S,7S,10S)-10-((S)-2-(((Benzyloxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)-N-methylbutanamido)-1 6 ,2 6-Methyl bis(2-((tert-butoxycarbonyl)amino)ethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclotetradecane-4-carboxylate (I-9)

[0195] Prepared according to the method of patent [CN111386283].

[0196] (f)(4S,7S,10S)-10-((S)-2-(((Benzyloxy)carbonyl)amino)-4-((tert-butoxycarbonyl)amino)-N-methylbutanamido)-1 6 ,2 6 -Bis(2-((tert-butoxycarbonyl)amino)ethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxylic acid (I-10)

[0197] At room temperature, I-9 (3.00 g, 2.87 mmol) was added to 30 ml of tetrahydrofuran and stirred to dissolve it. Then 5 ml of lithium hydroxide (481 mg, ) aqueous solution was added, and the reaction was carried out at room temperature for 1 h. The reaction was monitored by TLC until completion. Water was added, and the pH was adjusted to 3 with 1 M HCl aqueous solution. Ethyl acetate was used for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 2.74 g of white solid with a yield of 92%. 1 1H NMR (400 MHz, DMSO-d6) δ 12.91 (s, 1H), 8.89 (d, J = 7.9 Hz, 1H), 8.41 (d, J = 9.1 Hz, 1H), 7.63 (d, J = 7.7 Hz, 1H), 7.39 - 7.37 (m, 3H), 7.34 - 7.31 (m, 3H), 7.24 - 7.17 (m, 2H), 7.09 - 6.98 (m, 3H), 6.72 - 6.64 (m, 3H), 6.30 (s, 1H), 5.06 (s, 1H), 4.76 - 4.64 (m, 2H), 4.50 (s, 1H), 4.03 - 3.94 (m, 4H), 3.27 - 3.11 (m, 5H), 3.06 - 2.90 (m, 3H), 2.71 (s, 3H), 1.90 - 1.77 (m, 1H), 1.66 - 1.54 (m, 1H), 1.36 - 1.32 (m, 27H), 1.17 (d, J = 6.9 Hz, 3H).

[0198] (g)((S)-4-(((3S,6S,9S)-1 6 ,2 6-Bis(2-((tert-butoxycarbonyl)amino)ethoxy)-9-((cyanomethyl)carbamoyl)-6-methyl-4,7-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecan-3-yl)(methyl)amino)-4-oxobutane-1,3-diyl)di-tert-butyl dicarbonate (I-11)

[0199] Under ice bath conditions, I-10 (2.44 g, 2.36 mmol), aminoacetonitrile hydrochloride (284 mg, 3.07 mmol) and N,N-diisopropylethylamine (763 mg, 5.90 mmol) were added to 15 ml of dichloromethane, stirred to dissolve, and 5 ml of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (1.17 g, 3.07 mmol) in N,N-dimethylformamide was added dropwise. The reaction was carried out at ice bath for 30 min. The reaction was monitored by TLC until complete. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography [dichloromethane:methanol = 30:1] to obtain 2.38 g of white solid with a yield of 94%. 1 H NMR (400 MHz, DMSO-d6) δ 9.00 (d, J = 7.6 Hz, 1H), 8.67 (t, J = 5.7 Hz, 1H), 8.36 (d, J = 9.0 Hz, 1H), 7.61 (d, J = 7.7 Hz, 1H), 7.44 - 7.23 (m, 6H), 7.21 - 7.14 (m, 1H), 7.07 - 7.01 (m, 2H), 6.98 - 6.88 (m, 1H), 6.77 - 6.59 (m, 4H), 6.30 (s, 1H), 5.05 (s, 2H), 4.79 - 4.62 (m, 2H), 4.44 - 4.34 (m, 1H), 4.21 - 4.10 (m, 2H), 4.06 - 3.91 (m, 4H), 3.25 - 3.11 (m, 5H), 3.07 - 2.88 (m, 3H), 2.71 (s, 3H), 1.88 - 1.72 (m, 1H), 1.67 - 1.53 (m, 1H), 1.37 - 1.30 (m, 27H), 1.18 (d, J = 6.6 Hz, 3H).

[0200] (h)(2-(((4S,7S,10S)-10-((S)-2-Amino-4-((tert-butoxycarbonyl)amino)-N-methylbutanamido)-2 6 -(2-((tert-butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecanyl-1 6 -yl)oxy)ethyl)amino tert-butyl carbonate (I-12)

[0201] Under room temperature conditions, I-11 (2.38 g, 2.22 mmol) was added to 80 ml of ethanol, stirred to dissolve it, one drop of ammonia water and 10% palladium on carbon (1 g) were added, and the reaction was carried out at room temperature for 2 h under a hydrogen atmosphere. The reaction was monitored by TLC until completion, filtered through filter paper, and the filtrate was concentrated under reduced pressure and purified by column chromatography [methylene chloride: methanol = 10:1] to obtain 1.25 g of a white solid with a yield of 60%. 1 H NMR (400 MHz, DMSO-d6) δ 9.08 - 8.98 (m, 1H), 8.72 - 8.61 (m, 1H), 8.44 - 8.28 (m, 1H), 7.22 - 7.16 (m, 1H), 7.11 - 7.06 (m, 2H), 7.05 - 7.00 (m, 1H), 6.96 - 6.86 (m, 1H), 6.73 - 6.63 (m, 4H), 6.30 (s, 1H), 4.78 - 4.66 (m, 2H), 4.21 - 4.14 (m, 2H), 4.03 - 3.96 (m, 4H), 3.23 - 2.99 (m, 9H), 2.68 (s, 3H), 1.92 - 1.76 (m, 1H), 1.75 - 1.59 (m, 1H), 1.36 - 1.33 (m, 27H), 1.18 (d, J = 6.6 Hz, 3H).

[0202] (i) (2 - (((4S,7S,10S)-10 - ((S)-4 - ((tert-butoxycarbonyl)amino)-N-methyl-2-(4-methyl-2-(4-(methyldithio)phenyl)pyrimidine-5-carboxamido)butanamido)-2 6 -(2 - ((tert-butoxycarbonyl)amino)ethoxy)-4 - ((cyanomethyl)aminocarbonyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-1 6 -yl)oxy)ethyl) carbamic acid tert-butyl ester (I-13-1)

[0203] Under ice bath conditions, I-8-1 (17 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) were added to 2 ml of N,N-dimethylformamide, stirred to dissolve them, N,N-diisopropylethylamine (14 mg, 0.11 mmol) was added, and the reaction was carried out at ice bath temperature for 10 min. The reaction was monitored by TLC until completion, water and ethyl acetate were added for extraction, the organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography [methylene chloride: methanol = 20:1] to obtain 44 mg of a white solid with a yield of 69%. 11H NMR (600 MHz, DMSO-d6) δ 9.03 (dd, J = 20.1, 7.4 Hz, 2H), 8.83 (s, 1H), 8.70 (t, J = 5.5 Hz, 1H), 8.46 (d, J = 8.2 Hz, 2H), 8.41 (d, J = 8.9 Hz, 1H), 7.72 (d, J = 8.4 Hz, 2H), 7.20 (d, J = 8.4 Hz, 1H), 7.12 (s, 2H), 7.04 (d, J = 8.1 Hz, 2H), 6.75 (s, 2H), 6.69 (d, J = 23.3 Hz, 2H), 6.37 (s, 1H), 4.86 - 4.80 (m, 1H), 4.76 - 4.68 (m, 2H), 4.18 (t, J = 5.2 Hz, 2H), 4.05 - 3.96 (m, 4H), 3.26 - 3.06 (m, 8H), 2.82 (s, 3H), 2.67 (s, 3H), 2.54 (s, 3H), 1.98 - 1.94 (m, 1H), 1.77 - 1.68 (m, 1H), 1.37 - 1.33 (m, 27H), 1.20 (d, J = 6.7 Hz, 3H).

[0204] (j) Compound 1

[0205] At room temperature, I-13-1 (38 mg, 0.03 mmol) was added to 5 ml of hexafluoroisopropanol, stirred to dissolve it, trifluoroacetic acid (53 mg, 0.45 mmol) was added, protected by argon, and the reaction was carried out at 35 °C for 2 h. The reaction was monitored by UPLC until completion, concentrated to dryness, and purified by preparative liquid chromatography (C 18 Reverse-phase preparative column; solvent A, 0.1% aqueous TFA solution; solvent B, methanol; gradient elution for 30 min, B increased from 2% to 90%), to obtain Compound 1, 24 mg of white solid, yield 80%, 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.8 Hz, 1H), 8.85 (s, 1H), 8.78 (t, J = 5.7 Hz, 1H), 8.46 (d, J = 8.4 Hz, 2H), 8.38 (d, J = 9.0 Hz, 1H), 8.04 - 7.90 (m, 10H), 7.73 (d, J = 8.5 Hz, 2H), 7.24 (d, J = 8.2 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.03 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.14 (m, 6H), 3.20 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.54 (s, 3H), 2.13 - 2.07 (m, 1H), 2.02 - 1.97 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.47, 170.16, 166.20, 165.82, 163.05, 159.15 (q, J = 32.2 Hz), 156.36, 155.16, 154.15, 140.58, 135.49, 135.34, 131.39, 130.94, 130.27, 129.83, 129.73, 129.30, 128.68, 128.63, 127.36, 126.85, 118.03, 117.39 (q, J = 297.8 Hz), 115.02, 114.79, 66.16, 65.96, 59.82, 51.40, 48.46, 48.26, 38.69, 38.68, 36.43, 34.07, 33.01, 29.45, 27.80, 23.21, 23.00, 19.85. HRMS (ESI): Anal. Calcd for C 44 H 54 N 11 O7S2[(M + H) + : 912.3644, found: 912.3644.

[0206] Example 2: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-(ethyldithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6-Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 2)

[0207]

[0208] (a) Ethyl S-methylsulfonylthioate (I-7-2)

[0209] Under ice bath conditions, ethanethiol I-6-2 (200 mg, 3.22 mmol) and triethylamine (489 mg, 4.83 mmol) were added to 5 ml of tetrahydrofuran. Methanesulfonyl chloride (480 mg, 4.19 mmol) was added, and the reaction was carried out at room temperature for 5 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with dilute hydrochloric acid, aqueous sodium bicarbonate solution, water, and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 208 mg of a colorless oil, with a yield of 46%. 1 H NMR (600 MHz, Chloroform-d) δ 3.33 (s, 3H), 3.20 (q, J = 7.4 Hz, 2H), 1.46 (t, J = 7.4 Hz, 3H).

[0210] (b) 2-[4-(Ethyl disulfide)phenyl]-4-methylpyrimidine-5-carboxylic acid (I-8-2)

[0211] Using I-5 (30 mg, 0.12 mmol), triethylamine (31 mg, 0.31 mmol), and I-7-2 (20 mg, 0.15 mmol) as raw materials, according to the synthesis method of I-8-1, 29 mg of a white solid was obtained, with a yield of 79%. 1 H NMR (600 MHz, DMSO-d6) δ 9.12 (s, 1H), 8.44 (d, J = 8.6 Hz, 2H), 7.71 (d, J = 8.6 Hz, 2H), 2.86 (q, J = 7.3 Hz, 2H), 2.80 (s, 3H), 1.27 (t, J = 7.3 Hz, 3H).

[0212] (c) (2-(((4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-2-(2-(4-(ethyl disulfide)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-2 6 -(2-((tert-Butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-1 6 -yl)oxy)ethyl)carbamic acid tert-butyl ester (I-13-2)

[0213] Using I-8-2 (17 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I-13-1, 32 mg of white solid was obtained with a yield of 49%. 1 H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 12.4, 7.4 Hz, 2H), 8.82 (s, 1H), 8.66 (t, J = 5.6 Hz, 1H), 8.44 (d, J = 8.5 Hz, 2H), 8.37 (d, J = 8.9 Hz, 1H), 7.73 (d, J = 8.5 Hz, 2H), 7.19 (d, J = 8.4 Hz, 1H), 7.12 (s, 2H), 7.04 (dd, J = 8.6, 3.2 Hz, 1H), 7.01 - 6.94 (m, 1H), 6.75 (s, 1H), 6.72 - 6.61 (m, 3H), 6.37 (s, 1H), 4.87 - 4.81 (m, 1H), 4.77 - 4.70 (m, 2H), 4.20 - 4.16 (m, 2H), 4.05 - 3.98 (m, 4H), 3.25 - 3.08 (m, 8H), 2.89 - 2.84 (m, 2H), 2.82 (s, 3H), 2.67 (s, 3H), 2.00 - 1.94 (m, 1H), 1.78 - 1.70 (m, 1H), 1.36 - 1.34 (m, 27H), 1.28 (t, J = 7.3 Hz, 3H), 1.20 (d, J = 6.7 Hz, 3H).

[0214] (d) Compound 2

[0215] Using I-13-2 (26 mg, 0.02 mmol) and trifluoroacetic acid (36 mg, 0.32 mmol) as raw materials, according to the synthesis method of Compound 1, Compound 2 was obtained, 25 mg of white solid with a yield of 90%. 11H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.2 Hz, 1H), 8.99 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.76 (t, J = 5.0 Hz, 1H), 8.44 (d, J = 8.1 Hz, 2H), 8.38 (d, J = 8.9 Hz, 1H), 8.00 - 7.86 (m, 10H), 7.74 (d, J = 8.2 Hz, 2H), 7.24 (d, J = 8.2 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.5 Hz, 1H), 6.74 (s, 2H), 6.38 (s, 1H), 5.03 - 4.96 (m, 1H), 4.79 - 4.70 (m, 2H), 4.24 - 4.14 (m, 6H), 3.21 - 2.99 (m, 8H), 2.89 - 2.86 (m, 2H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.07 (m, 1H), 2.03 - 1.98 (m, 1H), 1.28 (t, J = 7.1 Hz, 3H), 1.22 (d, J = 6.4 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.24, 171.93, 170.97, 169.67, 165.69, 165.30, 162.57, 158.56 (q, J = 31.5 Hz), 155.86, 154.66, 153.65, 140.80, 134.87, 134.78, 130.90, 130.44, 129.77, 129.37, 129.25, 128.70, 128.16, 128.14, 126.85, 126.29, 117.54, 116.93 (q, J = 297.6), 114.50, 114.28, 65.66, 65.47, 59.33, 50.88, 47.94, 47.75, 38.20, 38.17, 35.94, 33.58, 32.51, 32.07, 28.97, 27.29, 22.71, 19.36, 14.03. HRMS (ESI): Anal. Calcd for C 45 H 56 N 11 O7S2[(M + H) + : 926.3800, found: 926.3796.

[0216] Example 3: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(propyl disulfide)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 , 26 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 3)

[0217]

[0218] Using propanethiol as the starting material, according to the synthesis method of Compound 2, through four steps (the synthetic route is as above), Compound 3 was prepared, 22 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 8.99 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.76 (t, J = 5.4 Hz, 1H), 8.44 (d, J = 8.3 Hz, 2H), 8.38 (d, J = 8.9 Hz, 1H), 8.02 - 7.87 (m, 10H), 7.73 (d, J = 8.4 Hz, 2H), 7.24 (d, J = 8.1 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.4 Hz, 1H), 6.75 (s, 2H), 6.38 (s, 1H), 5.00 (dt, J = 13.6, 6.4 Hz, 1H), 4.79 - 4.68 (m, 2H), 4.23 - 4.15 (m, 6H), 3.19 - 2.98 (m, 8H), 2.85 (s, 3H), 2.84 - 2.79 (m, 2H), 2.66 (s, 3H), 2.13 - 2.07 (m, 1H), 2.02 - 1.98 (m, 1H), 1.69 - 1.63 (m, 2H), 1.22 (d, J = 6.6 Hz, 3H), 0.95 (t, J = 7.3 Hz, 3H). 1313C NMR(126MHz,DMSO-d6)δ172.24,171.94,170.97,169.65,165.69,165.32,162.55,158.50(q,J=32.3Hz),155.86,154.65,153.64,140.75,134.89,134.83,130.88,130.44,129.73,129.33,129.23,128.69,128.17,128.12,126.83,126.34,117.53,116.85(q,J=297.8Hz),114.52,114.26,65.65,65.46,59.31,50.89,47.95,47.74,40.04,38.19,38.17,35.93,33.57,32.52,28.95,27.29,22.71,21.67,19.35,12.68.HRMS(ESI):Anal.Calcd for C 46 H 58 N 11 O7S2[(M+H) + :940.3957,found:940.3955.

[0219] Example 4: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-(isopropylthio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 4)

[0220]

[0221] Using isopropyl mercaptan as the starting material, according to the synthesis method of Compound 2, through four steps (the synthesis route is as above), Compound 4 was prepared, 30 mg of white solid, 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.5 Hz, 1H), 8.43 (d, J = 8.4 Hz, 2H), 8.38 (d, J = 9.0 Hz, 1H), 8.03 - 7.89 (m, 10H), 7.74 (d, J = 8.5 Hz, 2H), 7.24 (d, J = 9.1 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.72 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.78 - 4.71 (m, 2H), 4.24 - 4.14 (m, 6H), 3.24 - 3.21 (m, 1H), 3.18 - 2.97 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.02 - 1.98 (m, 1H), 1.29 (d, J = 6.7 Hz, 6H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.25, 171.93, 170.97, 169.65, 165.69, 165.31, 162.57, 158.73 (q, J = 32.3 Hz), 155.86, 154.65, 153.64, 141.28, 134.84, 134.79, 130.90, 130.44, 129.76, 129.32, 129.23, 128.63, 128.18, 128.13, 126.80, 126.13, 117.53, 116.84 (q, J = 298.3), 114.51, 114.28, 65.66, 65.47, 59.31, 50.89, 47.94, 47.75, 41.31, 38.19, 35.93, 33.59, 32.51, 31.25, 28.98, 27.30, 22.71, 22.12, 19.36. HRMS (ESI): Anal. Calcd for C 46 H 58 N 11 O7S2[(M + H) + : 940.3957, found: 940.3957.

[0222] Example 5: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(butyldithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,26 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 5)

[0223]

[0224] Using n-butyl mercaptan as the starting material, following the synthesis method of Compound 2, Compound 5 was prepared through four steps (synthesis route as above), obtaining 32 mg of white solid 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.5 Hz, 1H), 8.44 (d, J = 8.5 Hz, 2H), 8.38 (d, J = 8.9 Hz, 1H), 8.01 - 7.88 (m, 10H), 7.73 (d, J = 8.5 Hz, 2H), 7.24 (d, J = 10.2 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.78 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 3.16 - 2.97 (m, 8H), 2.88 - 2.85 (m, 2H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.03 - 1.98 (m, 1H), 1.66 - 1.62 (m, 2H), 1.40 - 1.36 (m, 2H), 1.22 (d, J = 6.7 Hz, 3H), 0.85 (d, J = 7.4 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.74, 172.43, 171.47, 170.16, 166.20, 165.82, 163.06, 159.06 (q, J = 32.6 Hz), 156.35, 155.15, 154.14, 141.27, 135.39, 135.34, 131.40, 130.94, 130.26, 129.83, 129.72, 129.19, 128.67, 128.62, 127.33, 126.86, 118.03, 117.22 (q, J = 297.2 Hz), 115.01, 114.77, 66.15, 65.96, 59.81, 51.38, 48.44, 48.26, 38.71, 38.69, 38.30, 36.44, 33.01, 31.22, 30.83, 29.48, 27.80, 23.21, 21.34, 19.85, 13.91. HRMS (ESI): Anal. Calcd for C 47 H 60 N 11 O7S2[(M + H) + : 954.4113, found: 954.4109.

[0225] Example 6: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(tert-butyldithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 6)

[0226]

[0227] Using tert-butyl mercaptan as the starting material, following the synthetic method of Compound 2, through four steps (synthetic route as above), Compound 6 was prepared, 34 mg of white solid, 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 8.99 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.76 (t, J = 5.4 Hz, 1H), 8.41 (d, J = 8.4 Hz, 2H), 8.38 (d, J = 9.0 Hz, 1H), 8.01 - 7.88 (m, 10H), 7.76 (d, J = 8.5 Hz, 2H), 7.24 (d, J = 9.2 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.96 (m, 1H), 4.79 - 4.72 (m, 2H), 4.22 - 4.17 (m, 6H), 3.19 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.02 - 1.98 (m, 1H), 1.33 (s, 9H), 1.22 (d, J = 6.6 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.24, 171.92, 170.97, 169.65, 165.69, 165.31, 162.59, 158.56 (q, J = 33.5 Hz), 155.85, 154.65, 153.64, 141.64, 134.83, 134.70, 130.90, 130.44, 129.76, 129.32, 129.23, 128.53, 128.17, 128.12, 126.75, 126.03, 117.53, 116.44 (q, J = 295.5 Hz), 114.51, 114.27, 65.66, 65.47, 59.30, 50.87, 49.58, 47.93, 47.76, 38.21, 38.18, 35.94, 32.52, 29.35, 28.99, 28.68, 27.29, 22.71, 19.36. HRMS (ESI): Anal. Calcd for C 47 H 60 N 11 O7S2[(M + H) + : 954.4113, found: 954.4119.

[0228] Example 7: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(pentyl disulfide)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 , 2 6-Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 7)

[0229]

[0230] Using n-pentyl mercaptan as the starting material, following the synthesis method of Compound 2, through four steps (synthesis route as above), Compound 7 was prepared, 35 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.44 (d, J = 8.5 Hz, 2H), 8.38 (d, J = 8.9 Hz, 1H), 8.01 - 7.90 (m, 10H), 7.73 (d, J = 8.6 Hz, 2H), 7.24 (d, J = 8.6 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.78 - 4.71 (m, 2H), 4.23 - 4.15 (m, 6H), 3.16 - 2.97 (m, 8H), 2.86 - 2.83 (m, 5H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.01 - 1.97 (m, 1H), 1.66 - 1.63 (m, 2H), 1.36 - 1.32 (m, 2H), 1.28 - 1.25 (m, 2H), 1.22 (d, J = 6.7 Hz, 3H), 0.84 (d, J = 7.3 Hz, 3H). 1313C NMR(126MHz, DMSO-d6) δ 172.75, 172.45, 171.47, 170.16, 166.21, 165.82, 163.05, 159.16 (d, J = 32.3 Hz), 156.35, 155.15, 154.14, 141.28, 135.40, 135.34, 131.40, 130.93, 130.26, 129.83, 129.72, 129.18, 128.67, 128.62, 127.34, 126.91, 118.03, 117.47 (q, J = 299.2 Hz), 115.02, 114.77, 66.15, 65.96, 59.82, 51.39, 48.46, 48.26, 38.70, 38.68, 38.62, 36.43, 34.06, 33.01, 30.29, 29.45, 28.40, 27.80, 23.21, 22.12, 19.85, 14.20. HRMS(ESI): Anal. Calcd for C 48 H 62 N 11 O7S2[(M + H) + : 968.4270, found: 968.4273.

[0231] Example 8: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-(cyclopentyl dithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 8)

[0232]

[0233] Using cyclopentyl mercaptan as the starting material, according to the synthesis method of Compound 2, after four steps (synthesis route as above), Compound 8 was obtained as a white solid, 34 mg, 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.4 Hz, 1H), 8.43 (d, J = 8.4 Hz, 2H), 8.38 (d, J = 8.9 Hz, 1H), 8.04 - 7.92 (m, 10H), 7.74 (d, J = 8.5 Hz, 2H), 7.24 (d, J = 7.6 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.03 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 3.18 - 2.99 (m, 8H), 2.99 - 2.92 (m, 1H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.02 - 1.95 (m, 3H), 1.72 - 1.67 (m, 2H), 1.63 - 1.55 (m, 4H), 1.22 (d, J = 6.6 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.25, 171.96, 170.98, 169.66, 165.72, 165.32, 162.57, 158.74 (q, J = 32.8 Hz), 155.85, 154.65, 153.64, 141.06, 134.90, 134.84, 130.89, 130.43, 129.77, 129.34, 129.21, 128.67, 128.17, 128.13, 126.81, 126.18, 117.53, 116.91 (q, J = 297.6 Hz), 114.53, 114.28, 65.65, 65.45, 59.34, 50.91, 49.81, 47.97, 47.77, 38.20, 38.19, 35.93, 33.57, 32.51, 32.22, 28.95, 27.29, 24.17, 22.71, 19.34. HRMS (ESI): Anal. Calcd for C 48 H 60 N 11 O7S2 [(M + H) + : 966.4113, found: 966.4115.

[0234] Example 9: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-((methyldisulfanyl)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclotetradecane-4-carboxamide trifluoroacetate (Compound 9)

[0235]

[0236] (a) Ethyl 2-(4-(hydroxymethyl)phenyl)-4-methylpyrimidine-5-carboxylate (I-15-9)

[0237] At room temperature, (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol (1.00 g, 4.27 mmol) and I-3 (854 mg, 4.27 mmol) were added to 30 ml of 1,4-dioxane, stirred to dissolve, 10 ml of aqueous sodium carbonate (905 mg, 8.54 mmol) solution and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (312 mg, 0.43 mmol) were added successively, protected by argon, heated to 90 °C and reacted for 1 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography [petroleum ether:ethyl acetate = 8:1] to obtain 809 mg of white solid with a yield of 70%. 1 1H NMR (400 MHz, Chloroform-d) δ 9.22 (s, 1H), 8.53 (d, J = 8.3 Hz, 2H), 7.50 (d, J = 7.9 Hz, 2H), 4.79 (s, 2H), 4.43 (q, J = 7.1 Hz, 2H), 2.91 (s, 3H), 1.43 (t, J = 7.1 Hz, 3H).

[0238] (b) Ethyl 2-[4-((acetylthio)methyl)phenyl]-4-methylpyrimidine-5-carboxylate (I-16-9)

[0239] Under ice bath conditions, triphenylphosphine (2.33 g, 8.88 mmol) was added to 10 ml of tetrahydrofuran, stirred to dissolve it, protected by argon, and diisopropyl azodicarboxylate (1.79 g, 8.88 mmol) was slowly added dropwise. After reacting for 20 min under ice bath, 4 ml of a mixed solution of I-15-9 (805 mg, 2.96 mmol) and thioacetic acid (8.88 mmol) in tetrahydrofuran was added, and the reaction was carried out for 30 min under ice bath. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography [petroleum ether: ethyl acetate = 25:1] to obtain 787 mg of a white solid with a yield of 81%. 1 H NMR (600 MHz, Chloroform-d) δ 9.20 (s, 1H), 8.45 (d, J = 8.3 Hz, 2H), 7.42 (d, J = 8.1 Hz, 2H), 4.42 (q, J = 7.1 Hz, 2H), 4.18 (s, 2H), 2.89 (s, 3H), 2.37 (s, 3H), 1.43 (t, J = 7.1 Hz, 3H).

[0240] (c) 2-[4-(Mercaptomethyl)phenyl]-4-methylpyrimidine-5-carboxylic acid (I-17-9)

[0241] Under room temperature conditions, I-16-9 (780 mg, 2.36 mmol) was added to 5 ml of methanol, stirred to dissolve it, 5 ml of an aqueous solution of sodium hydroxide (378 mg, 9.45 mmol) was added, protected by argon, and the temperature was raised to 50 °C and reacted for 1 h. The reaction was monitored by TLC until completion. Water was added, and the pH was adjusted to 3 with 1 M aqueous HCl solution. Ethyl acetate was added for extraction. The organic layers were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 615 mg of a white solid with a yield of 99%. 1 H NMR (400 MHz, DMSO-d6) δ 13.54 (s, 1H), 9.16 (s, 1H), 8.40 (d, J = 8.4 Hz, 2H), 7.53 (d, J = 8.4 Hz, 2H), 3.83 (d, J = 7.9 Hz, 2H), 2.99 (t, J = 7.9 Hz, 1H), 2.82 (s, 3H).

[0242] (d) 4-Methyl-2-(4-((methyldithio)methyl)phenyl)pyrimidine-5-carboxylic acid (I-18-9)

[0243] At room temperature, I-17-9 (25 mg, 0.10 mmol) and triethylamine (29 mg, 0.29 mmol) were added to 2 ml of methanol, stirred to dissolve, methyl methanesulfonate (15 mg, 0.12 mmol) was added, and the reaction was carried out at room temperature for 30 min. The reaction was monitored by TLC until completion. Water was added, and the pH was adjusted to 3 with 1 M aqueous HCl solution. Ethyl acetate was added for extraction. The organic layers were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Column chromatography [dichloromethane:methanol = 10:1] gave 22 mg of a white solid with a yield of 70%. 1 1H NMR (600 MHz, DMSO-d6) δ 8.91 (s, 1H), 8.35 (d, J = 7.9 Hz, 2H), 7.48 (d, J = 8.0 Hz, 2H), 4.06 (s, 2H), 2.75 (s, 3H), 2.20 (s, 3H).

[0244] (e)(2 - (((4S,7S,10S)-10 - ((S)-4 - ((tert-butoxycarbonyl)amino)-N-methyl-2-(4-methyl-2-(4-((methyldithio)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-2 6 -(2 - ((tert-butoxycarbonyl)amino)ethoxy)-4 - ((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-1 6 -yl)oxy)ethyl) tert-butyl carbamate (I-19-9)

[0245] Under ice bath conditions, I-18-9 (24 mg, 0.08 mmol), I-12 (50 mg, 0.05 mmol) and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) were added to 2 ml of N,N-dimethylformamide, stirred to dissolve, N,N-diisopropylethylamine (14 mg, 0.11 mmol) was added, and the reaction was carried out under ice bath for 10 min. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Column chromatography [dichloromethane:methanol = 20:1] gave 37 mg of a white solid with a yield of 57%. 11H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 15.6, 7.2 Hz, 2H), 8.83 (s, 1H), 8.67 (t, J = 5.5 Hz, 1H), 8.41 (d, J = 8.0 Hz, 2H), 8.37 (d, J = 8.7 Hz, 1H), 7.55 (d, J = 8.1 Hz, 2H), 7.20 (d, J = 8.4 Hz, 1H), 7.12 (s, 2H), 7.04 (d, J = 8.6 Hz, 1H), 7.01 - 6.96 (m, 1H), 6.75 (s, 1H), 6.72 - 6.62 (m, 3H), 6.37 (s, 1H), 4.87 - 4.81 (m, 1H), 4.78 - 4.71 (m, 2H), 4.20 - 4.16 (m, 2H), 4.09 (s, 2H), 4.06 - 4.02 (m, 2H), 4.00 - 3.97 (m, 2H), 3.24 - 3.08 (m, 8H), 2.83 (s, 3H), 2.67 (s, 3H), 2.25 - 2.22 (m, 3H), 2.00 - 1.95 (m, 1H), 1.78 - 1.70 (m, 1H), 1.37 - 1.34 (m, 27H), 1.20 (d, J = 6.7 Hz, 3H).

[0246] (f) Compound 9

[0247] Using I-19-9 (32 mg, 0.03 mmol) and trifluoroacetic acid (52 mg, 0.45 mmol) as starting materials, according to the synthesis method of Compound 1, Compound 9 was prepared as a white solid (23 mg, yield 64%). 1 1H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.3 Hz, 1H), 9.01 (d, J = 7.2 Hz, 1H), 8.84 (s, 1H), 8.79 (t, J = 4.8 Hz, 1H), 8.42 - 8.38 (m, 3H), 8.08 - 7.94 (m, 10H), 7.55 (d, J = 7.9 Hz, 2H), 7.24 (d, J = 8.3 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.5 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.37 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.70 (m, 2H), 4.25 - 4.15 (m, 6H), 4.09 (s, 2H), 3.17 - 2.98 (m, 8H), 2.84 (s, 3H), 2.66 (s, 3H), 2.24 (s, 3H), 2.12 - 2.08 (m, 1H), 2.00 - 1.96 (m, 1H), 1.21 (d, J = 6.3 Hz, 3H). 1313C NMR (201 MHz, DMSO-d6) δ 172.75, 172.47, 171.48, 170.17, 166.24, 165.78, 163.44, 158.95 (d, J = 32.7 Hz), 156.34, 155.17, 154.16, 141.65, 135.98, 135.34, 131.39, 130.96, 130.28, 130.10, 129.83, 129.75, 128.69, 128.64, 128.55, 127.31, 118.04, 117.52 (q, J = 298.6 Hz), 115.06, 114.82, 66.15, 65.96, 59.83, 51.40, 48.48, 48.26, 41.37, 38.69, 38.66, 36.43, 35.57, 34.07, 33.02, 27.80, 23.22, 22.65, 19.85. HRMS (ESI): Anal. Calcd for C 45 H 56 N 11 O7S2[(M + H) + : 926.3800, found: 926.3801.

[0248] Example 10: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((ethyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 10)

[0249]

[0250] (a) 2-(4-((Ethyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxylic acid (I-18-10)

[0251] Using I-17-9 (30 mg, 0.12 mmol), triethylamine (35 mg, 0.35 mmol) and I-7-2 (22 mg, 0.12 mmol) as raw materials, according to the synthesis method of I-18-9, 35 mg of white solid was obtained with a yield of 95%. 11H NMR (600 MHz, Methanol-d4) δ 9.10 (s, 1H), 8.41 (d, J = 8.3 Hz, 2H), 7.48 (d, J = 8.2 Hz, 2H), 3.97 (s, 2H), 2.84 (s, 3H), 2.44 (q, J = 7.4 Hz, 2H), 1.19 (t, J = 7.4 Hz, 3H).

[0252] (b) (2 - (((4S,7S,10S)-10 - ((S)-4 - ((tert-Butoxycarbonyl)amino)-2-(2-(4 - ((ethyldithio)methyl)phenyl)-4 - methylpyrimidine-5 - carboxamido)-N - methylbutanamido)-26-(2 - ((tert-Butoxycarbonyl)amino)ethoxy)-4 - ((cyanomethyl)carbamoyl)-7 - methyl-6,9 - dioxo-5,8 - diaza-1,2(1,3)-dibenzocyclodecane-16 - yl)oxy)ethyl) tert-Butylcarbamate (I-19-10)

[0253] Using I-18-10 (18 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-Tetramethyl-O-(7 - azabenzotriazol-1 - yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N - Diisopropylethylamine (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I-19-9, 46 mg of white solid was obtained, with a yield of 70%. 1 1H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 15.7, 7.4 Hz, 2H), 8.82 (s, 1H), 8.66 (t, J = 5.6 Hz, 1H), 8.43 - 8.39 (m, 2H), 8.39 - 8.35 (m, 1H), 7.53 (d, J = 8.3 Hz, 2H), 7.20 (s, 1H), 7.12 (s, 2H), 7.04 (d, J = 8.6 Hz, 1H), 7.02 - 6.95 (m, 1H), 6.75 (s, 1H), 6.73 - 6.61 (m, 3H), 6.37 (s, 1H), 4.87 - 4.81 (m, 1H), 4.78 - 4.69 (m, 2H), 4.18 (t, J = 4.9 Hz, 2H), 4.07 - 4.02 (m, 4H), 4.01 - 3.97 (m, 2H), 3.27 - 3.07 (m, 8H), 2.83 (s, 3H), 2.67 (s, 3H), 2.55 (q, J = 6.5, 5.8 Hz, 2H), 1.99 - 1.93 (m, 1H), 1.77 - 1.71 (m, 1H), 1.38 - 1.34 (m, 27H), 1.20 - 1.17 (m, 6H).

[0254] (c) Compound 10

[0255] Using I-19-10 (41 mg, 0.03 mmol) and trifluoroacetic acid (57 mg, 0.50 mmol) as starting materials, according to the synthesis method of Compound 1, Compound 10 was prepared as a white solid (25 mg, yield 54%). 1 H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.41 - 8.37 (m, 3H), 8.05 - 7.91 (m, 10H), 7.53 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 8.5 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.78 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 4.06 (s, 2H), 3.19 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.56 (q, J = 7.2 Hz, 2H), 2.13 - 2.09 (m, 1H), 2.03 - 1.98 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H), 1.19 (t, J = 7.3 Hz, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 172.24, 171.94, 170.96, 169.65, 165.73, 165.26, 162.92, 158.49 (q, J = 31.5 Hz), 155.82, 154.65, 153.64, 141.15, 135.45, 134.82, 130.88, 130.44, 129.89, 129.74, 129.31, 129.23, 128.17, 128.12, 128.02, 126.80, 117.53, 116.92 (q, J = 298.1 Hz), 114.52, 114.29, 65.65, 65.46, 59.31, 50.88, 47.95, 47.76, 41.70, 38.20, 38.17, 35.93, 33.56, 32.52, 31.20, 28.96, 27.30, 22.72, 19.35, 14.11. HRMS (ESI): Anal. Calcd for C 46 H 58 N 11 O7S2[(M + H) +:940.3957, found:940.3954.

[0256] Example 11: (4S,7S,10S)-10-((S)-4-amino-N-methyl-2-(4-methyl-2-(4-((propyl disulfanyl)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 11)

[0257]

[0258] Using I-17-9 and I-7-3 as starting materials, following the synthetic method of Compound 10, through three steps (synthetic route as above), Compound 11 was prepared, 26 mg of white solid, 1 1H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.79 (t, J = 5.5 Hz, 1H), 8.41 - 8.37 (m, 3H), 8.09 - 7.91 (m, 10H), 7.53 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 8.6 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.37 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.25 - 4.16 (m, 6H), 4.06 (s, 2H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.53 - 2.52 (m, 2H), 2.14 - 2.09 (m, 1H), 2.03 - 1.98 (m, 1H), 1.58 (q, J = 7.3 Hz, 2H), 1.22 (d, J = 6.7 Hz, 3H), 0.88 (t, J = 7.3 Hz, 3H). 1313C NMR(126MHz,DMSO-d6)δ172.74,172.46,171.47,170.16,166.25,165.77,163.42,159.00(q,J=32.3Hz),156.33,155.15,154.14,141.65,135.96,135.33,131.39,131.37,130.94,130.24,129.83,129.72,128.67,128.63,128.52,127.31,118.04,117.61(q,J=297.4Hz),115.05,114.82,66.13,65.95,59.83,51.40,48.48,48.26,42.15,38.68,38.66,36.42,34.06,33.02,32.12,29.41,27.79,23.22,22.13,19.84,13.25.HRMS(ESI):Anal.Calcd for C 47 H 60 N 11 O7S2[(M+H) + :954.4113,found:954.4113.

[0259] Example 12: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((isopropyl disulfanyl)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 12)

[0260]

[0261] Using I-17-9 and I-7-4 as starting materials, according to the synthesis method of Compound 10, through three steps (synthesis route as above), Compound 12 was prepared, 27 mg of white solid, 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.42 - 8.37 (m, 3H), 8.03 - 7.89 (m, 10H), 7.53 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 9.3 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 4.06 (s, 2H), 3.17 - 2.97 (m, 8H), 2.92 - 2.87 (m, 1H), 2.85 (s, 3H), 2.66 (s, 3H), 2.14 - 2.09 (m, 1H), 2.01 - 1.97 (m, 1H), 1.23 - 1.21 (m, 9H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.74, 172.44, 171.46, 170.16, 166.23, 165.76, 163.41, 158.99 (q, J = 32.7 Hz), 156.32, 155.15, 154.14, 141.59, 135.96, 135.34, 131.38, 130.94, 130.22, 130.10, 129.82, 129.73, 128.68, 128.62, 128.52, 127.31, 118.03, 117.34 (q, J = 297.8 Hz), 115.02, 114.80, 66.15, 65.96, 59.82, 51.39, 48.45, 48.26, 43.11, 38.69, 38.67, 36.43, 34.07, 33.02, 29.47, 29.04, 27.80, 23.22, 22.76, 19.86. HRMS (ESI): Anal. Calcd for C 47 H 60 N 11 O7S2[(M + H) + : 954.4113, found: 954.4108.

[0262] Example 13: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((butyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6-Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 13)

[0263]

[0264] Using I-17-9 and I-7-5 as starting materials, following the synthesis method of Compound 10, through three steps (the synthetic route is as above), Compound 13 was obtained as a white solid (23 mg). 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.41 - 8.37 (m, 3H), 8.04 - 7.91 (m, 10H), 7.53 (d, J = 8.2 Hz, 2H), 7.24 (d, J = 9.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.72 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.79 - 4.73 (m, 2H), 4.23 - 4.15 (m, 6H), 4.06 (s, 2H), 3.17 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.54 - 2.53 (m, 2H), 2.14 - 2.09 (m, 1H), 2.00 - 1.97 (m, 1H), 1.53 - 1.50 (m, 2H), 1.30 - 1.28 (m, 2H), 1.22 (d, J = 6.7 Hz, 3H), 0.82 (d, J = 7.5 Hz, 3H). 1313C NMR(126MHz,DMSO-d6)δ172.74,172.44,171.47,170.15,166.23,165.74,163.42,159.06(q,J=32.3Hz),156.32,155.15,154.14,141.68,135.97,135.33,131.39,130.93,130.23,130.09,129.85,129.73,128.67,128.62,128.51,127.30,118.03,117.48(q,J=299.3Hz),115.02,114.80,66.15,65.96,59.81,51.38,48.45,48.25,42.18,38.69,38.67,37.49,36.42,34.09,33.02,30.96,29.45,27.79,23.22,21.41,19.85,13.89.HRMS(ESI):Anal.Calcd for C 48 H 62 N 11 O7S2[(M+H) + :968.4270,found:968.4270.

[0265] Example 14: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((isobutyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 14)

[0266]

[0267] (a) Isobutyl S-methylsulfonylsulfate (I-7-14)

[0268] Using isobutyl mercaptan (200 mg, 2.22 mmol), triethylamine (337 mg, 3.33 mmol) and methanesulfonyl chloride (278 mg, 2.44 mmol) as raw materials, according to the synthesis method of I-7-2, 268 mg of colorless oil was obtained with a yield of 72%. 11H NMR (600 MHz, Chloroform-d) δ 3.31 (s, 3H), 3.07 (d, J = 6.9 Hz, 2H), 2.01 (dt, J = 13.4, 6.7 Hz, 1H), 1.04 (d, J = 6.7 Hz, 6H).

[0269] (b) 2-(4-((Isobutyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxylic acid (I-18-14)

[0270] Using I-17-9 (30 mg, 0.12 mmol), triethylamine (35 mg, 0.35 mmol) and I-7-14 (23 mg, 0.14 mmol) as starting materials, according to the synthesis method of I-18-9, 33 mg of white solid was obtained, with a yield of 82%. 1 1H NMR (400 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.40 (d, J = 8.3 Hz, 2H), 7.51 (d, J = 8.3 Hz, 2H), 4.04 (s, 2H), 2.80 (s, 3H), 2.39 (d, J = 6.8 Hz, 2H), 1.79 (dt, J = 13.4, 6.7 Hz, 1H), 0.87 (d, J = 6.7 Hz, 6H).

[0271] (c) (2-(((4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-2-(2-(4-((isobutyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-2 6 -(2-((tert-Butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-1 6 -yl)oxy)ethyl) tert-butyl carbamate (I-19-14)

[0272] Using I-18-14 (20 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as starting materials, according to the synthesis method of I-19-9, 47 mg of white solid was obtained, with a yield of 65%. 11H NMR (600 MHz, DMSO-d6) δ 8.98 (dd, J = 14.3, 7.4 Hz, 2H), 8.81 (s, 1H), 8.66 (t, J = 5.6 Hz, 1H), 8.40 (d, J = 8.1 Hz, 2H), 8.38 - 8.35 (m, 1H), 7.52 (d, J = 7.9 Hz, 2H), 7.19 (d, J = 8.4 Hz, 1H), 7.11 (s, 2H), 7.03 (d, J = 8.6 Hz, 1H), 7.00 - 6.95 (m, 1H), 6.75 (s, 1H), 6.72 - 6.61 (m, 3H), 6.36 (s, 1H), 4.86 - 4.80 (m, 1H), 4.77 - 4.69 (m, 2H), 4.17 (t, J = 5.0 Hz, 2H), 4.05 - 4.01 (m, 4H), 3.98 (s, 2H), 3.26 - 3.18 (m, 4H), 3.14 - 2.96 (m, 4H), 2.82 (s, 3H), 2.66 (s, 3H), 2.40 (d, J = 6.8 Hz, 2H), 1.98 - 1.93 (m, 1H), 1.84 - 1.78 (m, 1H), 1.76 - 1.68 (m, 1H), 1.36 - 1.33 (m, 27H), 1.19 (d, J = 6.7 Hz, 3H), 0.87 (d, J = 6.6 Hz, 6H).

[0273] (d) Compound 14

[0274] Using I-19-14 (41 mg, 0.03 mmol) and trifluoroacetic acid (55 mg, 0.48 mmol) as starting materials, according to the synthesis method of Compound 1, Compound 14 was prepared as a white solid (33 mg, yield 72%). 11H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.5 Hz, 1H), 8.41 - 8.38 (m, 3H), 8.03 - 7.90 (m, 10H), 7.54 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 8.5 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.72 (m, 2H), 4.24 - 4.15 (m, 6H), 4.06 (s, 2H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.42 (d, J = 6.8 Hz, 2H), 2.14 - 2.09 (m, 1H), 2.03 - 1.99 (m, 1H), 1.84 - 1.79 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H), 0.89 (d, J = 6.7 Hz, 6H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.74, 172.46, 171.47, 170.16, 166.26, 165.76, 163.42, 159.04 (q, J = 32.7 Hz), 156.32, 155.15, 154.14, 141.68, 135.97, 135.33, 131.38, 130.93, 130.25, 129.96, 129.82, 129.72, 128.67, 128.62, 128.51, 127.32, 118.03, 117.51 (q, J = 299.1 Hz), 115.02, 114.78, 66.15, 65.95, 59.82, 51.41, 48.46, 48.26, 46.97, 42.09, 38.69, 38.67, 36.43, 34.07, 33.01, 32.10, 29.44, 27.82, 23.21, 21.80, 19.84. HRMS (ESI): Anal. Calcd for C 48 H 62 N 11 O7S2[(M + H) + : 968.4270, found: 968.4271.

[0275] Example 15: (4S,7S,10S)-10-((S)-4-amino-2-(2-(4-((tert-butyldisulfanyl)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 15)

[0276]

[0277] Using I-17-9 and I-7-6 as starting materials, following the synthetic method of Compound 10, through three steps (synthetic route as above), Compound 15 was prepared, 31 mg of white solid, 1 1H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.5 Hz, 1H), 9.01 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.42 - 8.38 (m, 3H), 8.05 - 7.92 (m, 10H), 7.51 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 7.0 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 4.07 (s, 2H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.12 - 2.07 (m, 1H), 2.04 - 1.98 (m, 1H), 1.33 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.25, 171.95, 170.96, 169.66, 165.74, 165.26, 162.90, 158.64 (q, J = 31.1 Hz), 155.82, 154.65, 153.64, 140.88, 135.50, 134.83, 130.89, 130.43, 129.76, 129.70, 129.60, 129.31, 129.23, 128.18, 128.10, 126.81, 117.53, 116.98 (q, J = 297.1 Hz), 114.52, 114.29, 65.66, 65.47, 59.32, 50.90, 47.95, 47.92, 47.76, 43.79, 38.19, 38.18, 35.93, 33.57, 32.51, 29.67, 28.96, 27.30, 22.72, 19.35. HRMS (ESI): Anal. Calcd for C 48 H 62 N 11 O7S2[(M + H) + : 968.4270, found: 968.4273.

[0278] Example 16: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((cyclobutyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 16)

[0279]

[0280] (a) S-Cyclobutyl-4-methylbenzenesulfonyl sulfate (I-22-16)

[0281] At room temperature, potassium p-toluenethiosulfonate (120 mg, 0.53 mmol) was added to 3 ml of N,N-dimethylformamide, stirred to dissolve, and iodocyclobutane (96 mg, 0.53 mmol) was added. The reaction was carried out at room temperature for 12 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Column chromatography [petroleum ether:ethyl acetate = 15:1] gave 40 mg of a white solid with a yield of 31%. 11H NMR (600 MHz, Chloroform-d) δ 7.82 (d, J = 8.3 Hz, 2H), 7.35 (d, J = 8.1 Hz, 2H), 3.97 (p, J = 8.2 Hz, 1H), 2.47 (s, 3H), 2.38 - 2.32 (m, 2H), 2.12 - 2.04 (m, 2H), 2.00 - 1.93 (m, 2H).

[0282] (b) 2-(4-((Cyclobutyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxylic acid (I-18-16)

[0283] Using I-17-9 (30 mg, 0.12 mmol), triethylamine (35 mg, 0.35 mmol) and I-22-16 (34 mg, 0.14 mmol) as raw materials, according to the synthesis method of I-18-9, 34 mg of white solid was obtained, with a yield of 85%. 1 1H NMR (600 MHz, DMSO-d6) δ 9.04 (s, 1H), 8.39 (d, J = 8.3 Hz, 2H), 7.48 (d, J = 8.3 Hz, 2H), 4.03 (s, 2H), 3.51 (p, J = 8.0 Hz, 1H), 2.78 (s, 3H), 2.18 - 2.11 (m, 2H), 2.00 - 1.94 (m, 2H), 1.84 - 1.77 (m, 2H).

[0284] (c) (2-(((4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-2-(2-(4-((cyclobutyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-2 6 -(2-((tert-Butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-1 6 -yl)oxy)ethyl) tert-butyl carbamate (I-19-16)

[0285] Using I-18-16 (19 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I-19-9, 51 mg of white solid was obtained, with a yield of 76%. 11H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 15.4, 7.4 Hz, 2H), 8.82 (s, 1H), 8.67 (t, J = 5.6 Hz, 1H), 8.41 (d, J = 8.2 Hz, 2H), 8.37 (d, J = 9.1 Hz, 1H), 7.51 (d, J = 8.2 Hz, 2H), 7.20 (d, J = 8.2 Hz, 1H), 7.12 (s, 2H), 7.04 (d, J = 8.6 Hz, 1H), 7.01 - 6.98 (m, 1H), 6.75 (s, 1H), 6.72 - 6.63 (m, 3H), 6.37 (s, 1H), 4.86 - 4.82 (m, 1H), 4.76 - 4.71 (m, 2H), 4.18 (t, J = 4.9 Hz, 2H), 4.05 - 3.97 (m, 6H), 3.53 (q, J = 7.9 Hz, 1H), 3.27 - 3.07 (m, 8H), 2.83 (s, 3H), 2.67 (s, 3H), 2.19 - 2.10 (m, 2H), 2.02 - 1.93 (m, 4H), 1.85 - 1.80 (m, 1H), 1.77 - 1.70 (m, 1H), 1.37 - 1.34 (m, 27H), 1.20 (d, J = 6.8 Hz, 3H).

[0286] (c) Compound 16

[0287] Using I-19-16 (47 mg, 0.04 mmol) and trifluoroacetic acid (68 mg, 0.60 mmol) as starting materials, following the synthesis method of Compound 1, Compound 16 was prepared as a white solid (24 mg, yield 44%). 11H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.7 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.5 Hz, 1H), 8.42 - 8.37 (m, 3H), 8.01 - 7.89 (m, 10H), 7.52 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 8.6 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.78 - 4.71 (m, 2H), 4.23 - 4.14 (m, 6H), 4.05 (s, 2H), 3.57 - 3.53 (m, 1H), 3.17 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.19 - 2.14 (m, 2H), 2.12 - 2.06 (m, 1H), 2.01 - 1.96 (m, 3H), 1.86 - 1.79 (m, 2H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.74, 172.44, 171.46, 170.16, 166.24, 165.75, 163.41, 159.06 (q, J = 31.9 Hz), 156.31, 155.14, 154.14, 141.60, 135.96, 135.34, 131.39, 130.93, 130.28, 130.22, 129.82, 129.72, 128.67, 128.62, 128.54, 127.31, 118.03, 117.50 (q, J = 297.6 Hz), 115.00, 114.78, 66.15, 65.95, 59.82, 51.39, 48.44, 48.26, 43.84, 43.03, 38.70, 38.68, 36.43, 34.06, 33.01, 29.91, 29.46, 27.80, 23.21, 19.85, 17.93. HRMS (ESI): Anal. Calcd for C 48 H 60 N 11 O7S2[(M + H) + : 966.4113, found: 966.4120.

[0288] Example 17: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(pentyl disulfide)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 17)

[0289]

[0290] Using I-17-9 and I-7-7 as starting materials, according to the synthesis method of Compound 10, after three steps (synthesis route as above), Compound 17 was prepared, 30 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.6 Hz, 1H), 8.41 - 8.37 (m, 3H), 8.03 - 7.91 (m, 10H), 7.53 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 10.2 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.74 (m, 2H), 6.38 (s, 1H), 5.03 - 4.96 (m, 1H), 4.78 - 4.71 (m, 2H), 4.23 - 4.14 (m, 6H), 4.06 (s, 2H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.50 - 2.48 (m, 2H), 2.13 - 2.09 (m, 1H), 2.04 - 1.99 (m, 1H), 1.55 - 1.51 (m, 2H), 1.27 - 1.23 (m, 4H), 1.22 (d, J = 6.7 Hz, 3H), 0.83 (t, J = 7.0 Hz, 3H). 1313C NMR(126MHz, DMSO-d6) δ 172.75, 172.45, 171.48, 170.16, 166.25, 165.75, 163.43, 159.32(q, J = 32.1Hz), 156.31, 155.15, 154.14, 141.69, 135.98, 135.33, 131.39, 130.93, 130.27, 130.22, 129.83, 129.72, 128.68, 128.63, 128.52, 127.29, 118.03, 117.50(q, J = 297.8Hz), 115.02, 114.77, 66.16, 65.96, 59.83, 51.40, 48.45, 48.26, 42.20, 38.69, 38.68, 37.76, 36.44, 34.08, 33.01, 30.42, 29.46, 28.50, 27.80, 23.21, 22.08, 19.85, 14.25. HRMS(ESI): Anal. Calcd for C 49 H 64 N 11 O7S2[(M + H) + : 982.4426, found: 982.4427.

[0291] Example 18: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((cyclopentyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 18)

[0292]

[0293] Using I-17-9 and I-7-8 as starting materials, following the synthesis method of Compound 10, through three steps (synthesis route as above), Compound 18 was prepared, 24 mg of white solid 11H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 8.99 (d, J = 7.9 Hz, 1H), 8.83 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.41 - 8.37 (m, 3H), 8.02 - 7.90 (m, 10H), 7.52 (d, J = 8.3 Hz, 2H), 7.23 (d, J = 8.5 Hz, 1H), 7.18 (s, 2H), 7.09 (d, J = 8.6 Hz, 1H), 6.77 - 6.72 (m, 2H), 6.37 (s, 1H), 5.01 - 4.97 (m, 1H), 4.78 - 4.71 (m, 2H), 4.23 - 4.13 (m, 6H), 4.07 (s, 2H), 3.21 - 3.06 (m, 8H), 2.98 - 2.95 (m, 1H), 2.84 (s, 3H), 2.65 (s, 3H), 2.12 - 2.07 (m, 1H), 2.02 - 1.98 (m, 1H), 1.90 - 1.85 (m, 2H), 1.66 - 1.62 (m, 2H), 1.57 - 1.51 (m, 4H), 1.21 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.74, 172.45, 171.47, 170.16, 166.24, 165.75, 163.42, 159.10 (q, J = 32.8 Hz), 156.32, 155.15, 154.14, 141.62, 135.95, 135.33, 131.38, 130.93, 130.27, 130.22, 129.82, 129.72, 128.67, 128.62, 128.53, 127.31, 118.03, 117.49 (q, J = 295.8 Hz), 115.02, 114.79, 66.15, 65.96, 59.82, 51.40, 49.50, 48.46, 48.26, 42.64, 38.70, 38.68, 36.43, 34.07, 33.01, 32.92, 29.46, 27.80, 24.65, 23.21, 19.85. HRMS (ESI): Anal. Calcd for C 49 H 62 N 11 O7S2[(M + H) + : 980.4270, found: 980.4268.

[0294] Example 19: (4S,7S,10S)-10-((S)-4-amino-2-(2-(4-((heptyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 19)

[0295]

[0296] Using n-heptyl mercaptan and I-17-9 as starting materials, following the synthesis method of Compound 14, after four steps (synthesis route as above), Compound 19 was obtained as a white solid, 33 mg, 1 1H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.42 - 8.38 (m, 3H), 8.02 - 7.88 (m, 10H), 7.53 (d, J = 8.4 Hz, 2H), 7.24 (dd, J = 8.7, 1.9 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.72 (m, 2H), 6.38 (s, 1H), 5.03 - 4.98 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.14 (m, 6H), 4.05 (s, 2H), 3.19 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.50 - 2.47 (m, 2H), 2.13 - 2.08 (m, 1H), 2.02 - 1.98 (m, 1H), 1.53 - 1.48 (m, 2H), 1.26 - 1.24 (m, 2H), 1.22 (d, J = 6.8 Hz, 3H), 1.20 - 1.16 (m, 6H), 0.83 (t, J = 7.0 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.48, 170.17, 166.24, 165.75, 163.42, 159.27, 159.00, 156.31, 155.15, 154.15, 141.75, 135.97, 135.34, 131.40, 130.94, 130.28, 130.23, 129.83, 129.73, 128.68, 128.63, 128.52, 127.28, 118.03, 117.48 (q, J = 296.8 Hz), 115.02, 114.77, 66.16, 65.96, 59.83, 51.40, 48.45, 48.27, 42.24, 38.69, 37.79, 36.44, 34.07, 33.01, 31.59, 29.47, 28.83, 28.61, 28.21, 27.80, 23.22, 22.45, 19.85, 14.35. HRMS (ESI): Anal. Calcd for C 51 H 68 N 11 O7S2[(M + H) + : 1010.4739, found: 1010.4735.

[0297] Example 20: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((cyclohexyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-cyanomethyl-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 20)

[0298]

[0299] Using cyclohexanethiol and I-17-9 as starting materials, according to the synthesis method of Compound 14, through four steps (synthesis route as above), Compound 20 was prepared, 24 mg of white solid 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.79 (t, J = 5.5 Hz, 1H), 8.41 - 8.38 (m, 3H), 8.05 - 7.91 (m, 10H), 7.52 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 10.1 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.03 - 4.96 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 4.05 (s, 2H), 3.19 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.58 - 2.54 (m, 1H), 2.14 - 2.08 (m, 1H), 2.05 - 1.98 (m, 1H), 1.93 - 1.89 (m, 2H), 1.71 - 1.66 (m, 2H), 1.54 - 1.50 (m, 1H), 1.28 - 1.26 (m, 1H), 1.22 (d, J = 6.8 Hz, 3H), 1.20 - 1.12 (m, 4H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.68, 172.38, 171.41, 170.09, 166.19, 165.68, 163.37, 159.17 (q, J = 31.8 Hz), 156.25, 155.09, 154.08, 141.54, 135.93, 135.27, 131.32, 130.86, 130.17, 129.83, 129.76, 129.66, 128.61, 128.56, 128.42, 127.25, 117.96, 117.30 (q, J = 296.9 Hz), 114.96, 114.71, 66.09, 65.89, 59.76, 51.33, 48.69, 48.38, 48.20, 43.17, 38.63, 38.60, 36.36, 33.98, 32.95, 32.69, 29.39, 27.72, 25.85, 25.47, 23.15, 19.78. HRMS (ESI): Anal. Calcd for C 50 H 64 N 11 O7S2[(M + H) + : 994.4426, found: 994.4428.

[0300] Example 21: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(phenyldisulfanyl)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 21)

[0301]

[0302] (a) 4-Methyl-2-(4-((phenyldisulfanyl)methyl)phenyl)pyrimidine-5-carboxylic acid (I-18-21)

[0303] Using I-17-9 (25 mg, 0.10 mmol), triethylamine (29 mg, 0.29 mmol) and S-phenyl-4-methylbenzenesulfonyl thioester (32 mg, 0.12 mmol) as starting materials, according to the synthesis method of I-18-9, 26 mg of white solid was obtained, with a yield of 71%, 1 1H NMR (600 MHz, DMSO-d6) δ 9.12 (s, 1H), 8.37 (d, J = 8.2 Hz, 2H), 7.48 (d, J = 8.1 Hz, 4H), 7.34 (t, J = 7.7 Hz, 2H), 7.25 (t, J = 7.3 Hz, 1H), 4.13 (s, 2H), 2.80 (s, 3H).

[0304] (b) (2-(((4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-N-methyl-2-(4-methyl-2-(4-((phenyldisulfanyl)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-2 6 -(2-((tert-Butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecan-1 6 -yl)oxy)ethyl)carbamic acid tert-butyl ester (I-19-21)

[0305] Using I-18-21 (21 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as starting materials, according to the synthesis method of I-19-9, 45 mg of white solid was obtained, with a yield of 66%, 11H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 11.4, 7.4 Hz, 2H), 8.82 (s, 1H), 8.66 (t, J = 5.7 Hz, 1H), 8.37 (d, J = 8.2 Hz, 3H), 7.52 - 7.47 (m, 4H), 7.37 - 7.33 (m, 2H), 7.28 - 7.24 (m, 1H), 7.21 - 7.18 (m, 1H), 7.12 (s, 2H), 7.04 (d, J = 8.7 Hz, 1H), 7.01 - 6.97 (m, 1H), 6.75 (s, 1H), 6.72 - 6.62 (m, 3H), 6.37 (s, 1H), 4.88 - 4.81 (m, 1H), 4.77 - 4.70 (m, 2H), 4.20 - 4.13 (m, 4H), 4.05 - 4.02 (m, 2H), 4.01 - 3.96 (m, 2H), 3.26 - 3.08 (m, 8H), 2.83 (s, 3H), 2.67 (s, 3H), 1.99 - 1.94 (m, 1H), 1.78 - 1.69 (m, 1H), 1.37 - 1.34 (m, 27H), 1.20 (d, J = 6.8 Hz, 3H).

[0306] (c) Compound 21

[0307] Using I-19-21 (40 mg, 0.03 mmol) and trifluoroacetic acid (53 mg, 0.47 mmol) as starting materials, according to the synthesis method of Compound 1, Compound 21 was prepared as a white solid (23 mg, yield 51%). 1 1H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.40 - 8.35 (m, 3H), 8.03 - 7.89 (m, 10H), 7.52 - 7.49 (m, 4H), 7.37 - 7.34 (m, 2H), 7.29 - 7.23 (m, 2H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.72 (m, 2H), 6.38 (s, 1H), 5.03 - 4.98 (m, 1H), 4.79 - 4.70 (m, 2H), 4.27 - 4.17 (m, 6H), 4.15 (s, 2H), 3.17 - 2.97 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.02 - 1.98 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.48, 170.16, 166.25, 165.75, 163.37, 159.14 (q, J = 32.5 Hz), 156.32, 155.15, 154.15, 140.71, 136.61, 136.12, 135.34, 131.39, 130.93, 130.32, 129.93, 129.83, 129.72, 129.67, 128.67, 128.62, 128.58, 127.88, 127.60, 127.33, 118.04, 117.39 (q, J = 297.1 Hz), 115.02, 114.78, 66.16, 65.96, 59.82, 51.40, 48.45, 48.27, 42.29, 38.70, 38.69, 36.44, 34.07, 33.02, 29.46, 27.80, 23.21, 19.85. HRMS (ESI): Anal. Calcd for C 50 H 58 N 11 O7S2[(M + H) + : 988.3957, found: 988.3955.

[0308] Example 22: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(((benzyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 22)

[0309]

[0310] Using I-17-9 and S-benzyl methanethiosulfonate I-7-22 as starting materials, according to the synthesis method of Compound 10, through three steps (synthesis route as above), Compound 22 was prepared, 31 mg of white solid 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.80 (t, J = 5.6 Hz, 1H), 8.40 (t, J = 5.9 Hz, 3H), 8.07 - 7.94 (m, 10H), 7.46 (d, J = 8.4 Hz, 2H), 7.38 - 7.35 (m, 2H), 7.32 - 7.30 (m, 3H), 7.24 (dd, J = 8.6, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.37 (s, 1H), 5.01 - 4.97 (m, 1H), 4.77 - 4.70 (m, 2H), 4.25 - 4.15 (m, 6H), 3.85 (s, 2H), 3.81 (s, 2H), 3.18 - 2.98 (m, 8H), 2.84 (s, 3H), 2.66 (s, 3H), 2.14 - 2.09 (m, 1H), 2.04 - 1.99 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.69, 172.42, 171.41, 170.10, 166.20, 165.71, 163.34, 159.07 (q, J = 31.9 Hz), 156.25, 155.09, 154.08, 141.24, 137.66, 135.96, 135.28, 131.32, 130.86, 130.25, 129.99, 129.78, 129.64, 128.84, 128.67, 128.61, 128.57, 128.48, 127.75, 127.25, 117.96, 117.38 (q, J = 298.0 Hz), 114.97, 114.73, 66.07, 65.88, 59.78, 51.36, 48.43, 48.21, 42.02, 41.56, 38.63, 38.61, 36.36, 33.97, 32.95, 29.34, 27.73, 23.14, 19.77. HRMS (ESI): Anal. Calcd for C 51 H 60 N 11 O7S2[(M + H) + : 1002.4113, found: 1002.4115.

[0311] Example 23: (4S,7S,10S)-10-((S)-2-(2-(4-((allyldithio)methyl)phenyl)-4-methylpyrimidin-5-carboxamido)-4-amino-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 23)

[0312]

[0313] Using allyl mercaptan and I-17-9 as starting materials, according to the synthesis method of Compound 14, after four steps (the synthetic route is as above), Compound 23 was obtained as a white solid, 19 mg, 1 H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.41 (d, J = 8.2 Hz, 2H), 8.38 (d, J = 8.5 Hz, 1H), 8.05 - 7.90 (m, 10H), 7.53 (d, J = 8.3 Hz, 2H), 7.26 - 7.23 (m, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.81 - 5.74 (m, 1H), 5.15 - 5.09 (m, 2H), 5.02 - 4.97 (m, 1H), 4.78 - 4.70 (m, 2H), 4.24 - 4.15 (m, 6H), 4.07 (s, 2H), 3.21 (d, J = 7.4 Hz, 2H), 3.17 - 2.97 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.09 (m, 1H), 2.02 - 1.98 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.75, 172.46, 171.47, 170.16, 166.25, 165.77, 163.41, 159.05 (q, J = 32.5 Hz), 156.32, 155.15, 154.14, 141.49, 136.00, 135.33, 133.79, 131.38, 130.93, 130.31, 129.86, 129.83, 129.72, 128.67, 128.63, 128.53, 127.31, 119.25, 118.03, 117.56 (q, J = 297.4 Hz), 115.02, 114.79, 66.15, 65.95, 59.83, 51.41, 48.46, 48.26, 42.03, 40.90, 38.70, 38.68, 36.43, 33.01, 31.14, 29.45, 27.80, 23.21, 19.84. HRMS (ESI): Anal. Calcd for C 47 H 58 N 11 O7S2[(M + H) + : 952.3957, found: 952.3960.

[0314] Example 24: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(((2-Methoxyethyl)disulfanyl)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 24)

[0315]

[0316] Using potassium p-toluenethiosulfonate and 1-iodo-2-methoxyethane as starting materials, according to the synthesis method of Compound 16, after four steps (synthesis route as above), Compound 24 was prepared, 33 mg of white solid 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.42 - 8.37 (m, 3H), 8.05 - 7.90 (m, 10H), 7.54 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 8.3 Hz, 1H), 7.20 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.25 - 4.15 (m, 6H), 4.08 (s, 2H), 3.50 (t, J = 6.4 Hz, 2H), 3.24 (s, 3H), 3.17 - 2.97 (m, 8H), 2.85 (s, 3H), 2.75 (t, J = 6.3 Hz, 2H), 2.67 (s, 3H), 2.13 - 2.08 (m, 1H), 2.03 - 1.98 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.47, 170.16, 166.24, 165.76, 163.42, 159.22 (q, J = 31.2 Hz), 156.32, 155.15, 154.14, 141.55, 135.99, 135.34, 131.39, 130.94, 130.26, 129.92, 129.83, 129.73, 128.68, 128.63, 128.55, 127.31, 118.04, 117.56 (q, J = 298.5 Hz), 115.03, 114.79, 70.41, 66.15, 65.96, 59.82, 58.22, 51.40, 48.46, 48.26, 42.19, 38.70, 38.68, 37.70, 36.43, 34.06, 33.01, 29.45, 27.80, 23.21, 19.85. HRMS (ESI): Anal. Calcd for C 47 H 60 N 11 O8S2 [(M + H) + : 970.4062, found: 970.4060.

[0317] Example 25: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(((2-(methylthio)ethyl)disulfanyl)methyl)phenyl)pyrimidin-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 25)

[0318]

[0319] Using potassium p-toluenethiosulfonate and 2-chloroethyl methyl sulfide as starting materials, according to the synthesis method of Compound 16, after four steps (the synthetic route is as above), Compound 25 was prepared, 29 mg of white solid 1 H NMR(600MHz,DMSO-d6)δ9.19(d,J=7.3Hz,1H),9.00(d,J=7.8Hz,1H),8.84(s,1H),8.78(t,J=5.4Hz,1H),8.41(d,J=8.1Hz,2H),8.38(d,J=8.7Hz,1H),8.03-7.89(m,10H),7.55(d,J=8.2Hz,2H),7.24(d,J=8.2Hz,1H),7.20(s,2H),7.10(d,J=8.5Hz,1H),6.77-6.73(m,2H),6.38(s,1H),5.03-4.98(m,1H),4.78-4.71(m,2H),4.24-4.15(m,6H),4.09(s,2H),3.18-2.98(m,8H),2.85(s,3H),2.75-2.71(m,2H),2.67-2.65(m,5H),2.14-2.09(m,1H),2.01(s,3H),2.00-1.98(m,1H),1.22(d,J=6.6Hz,3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.48, 170.16, 166.25, 165.75, 163.41, 159.10 (q, J = 31.4 Hz), 156.32, 155.15, 154.15, 141.64, 136.02, 135.33, 131.39, 130.94, 130.27, 130.02, 129.83, 129.72, 128.68, 128.63, 128.56, 127.33, 118.03, 117.48 (q, J = 298.9 Hz), 115.03, 114.79, 66.15, 65.96, 59.82, 51.40, 48.46, 48.26, 42.24, 38.69, 38.67, 37.30, 36.43, 34.07, 33.01, 32.95, 29.44, 27.79, 23.21, 19.85, 14.96. HRMS (ESI): Anal. Calcd for C 47 H 60 N 11 O7S3[(M + H) + : 986.3834, found: 986.3832.

[0320] Example 26: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-((((trimethylsilyl)methyl)disulfanyl)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 26)

[0321]

[0322] Using potassium p-toluenethiosulfonate and iodomethyltrimethylsilane as starting materials, according to the synthesis method of Compound 16, after four steps (the synthetic route is as above), Compound 26 was prepared, 18 mg of white solid 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.7 Hz, 1H), 8.84 (s, 1H), 8.79 (t, J = 5.7 Hz, 1H), 8.41 - 8.37 (m, 3H), 8.25 - 7.79 (m, 10H), 7.53 (d, J = 8.0 Hz, 2H), 7.24 (d, J = 8.5 Hz, 1H), 7.20 (s, 2H), 7.10 (d, J = 8.5 Hz, 1H), 6.77 - 6.74 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.72 (m, 2H), 4.24 - 4.15 (m, 6H), 4.09 (s, 2H), 3.18 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.17 - 2.10 (m, 1H), 2.09 (s, 2H), 2.03 - 1.98 (m, 1H), 1.22 (d, J = 6.6 Hz, 3H), 0.05 (s, 9H). 13 13C NMR (126 MHz, DMSO-d6) δ 173.52, 173.23, 172.25, 170.93, 167.03, 166.52, 164.22, 159.82 (q, J = 31.1 Hz), 157.10, 155.93, 154.92, 142.33, 136.69, 136.11, 132.15, 131.71, 131.12, 131.05, 130.60, 130.50, 129.45, 129.41, 129.24, 128.07, 118.81, 118.36 (q, J = 297.6 Hz), 115.81, 115.58, 66.93, 66.74, 60.60, 52.18, 49.25, 49.04, 41.81, 39.47, 39.45, 37.20, 34.84, 33.80, 30.20, 28.57, 26.42, 24.00, 20.62, -0.49. HRMS (ESI): Anal. Calcd for C 48 H 64 N 11 O7S2Si[(M + H) + : 998.4195, found: 998.4196.

[0323] Example 27: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-(((3-Fluoropropyl)disulfanyl)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,26 -Bis(2-aminoethoxy)-N-cyanomethyl-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 27)

[0324]

[0325] Using potassium p-toluenethiosulfonate and 3-fluoro-1-iodopropane as starting materials, following the synthesis method of Compound 16, through four steps (synthesis route as above), Compound 27 was prepared, 29 mg of white solid 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.6 Hz, 1H), 8.42 - 8.37 (m, 3H), 8.06 - 7.90 (m, 10H), 7.54 (d, J = 8.4 Hz, 2H), 7.24 (d, J = 10.5 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.03 - 4.98 (m, 1H), 4.79 - 4.72 (m, 2H), δ 4.46 (dt, J = 47.4, 5.8 Hz, 2H), 4.24 - 4.14 (m, 6H), 4.08 (s, 2H), 3.19 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.65 - 2.62 (m, 2H), 2.14 - 2.08 (m, 1H), 2.03 - 2.00 (m, 1H), 1.97 - 1.91 (m, 2H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.74, 172.45, 171.47, 170.16, 166.25, 165.75, 163.41, 159.22 (q, J = 32.3 Hz), 156.31, 155.15, 154.14, 141.51, 136.01, 135.33, 131.38, 130.93, 130.24, 130.02, 129.82, 129.72, 128.67, 128.62, 128.55, 127.32, 118.03, 117.49 (q, J = 299.3 Hz), 115.02, 114.78, 82.62 (d, J = 162.9 Hz), 66.15, 65.96, 59.82, 51.40, 48.46, 48.26, 41.98, 38.69, 38.68, 36.43, 34.06, 33.40 (d, J = 4.6 Hz), 33.01, 29.81 (d, J = 19.8 Hz), 29.45, 27.79, 23.21, 19.84. HRMS (ESI): Anal. Calcd for C 47 H 59 FN 11 O7S2 [(M + H) + : 972.4019, found: 972.4021.

[0326] Example 28: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(((3,3-difluoropropyl)disulfanyl)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 28)

[0327]

[0328] (a) 3,3-Difluoropropyl 4-methylbenzenesulfonate (I-21-28)

[0329] Under ice bath conditions, 3,3-difluoro-1-propanol (100 mg, 0.52 mmol) and pyridine (411 mg, 5.21 mmol) were added to 2 ml of dichloromethane, stirred to dissolve, protected by argon, p-toluenesulfonyl chloride (119 mg, 0.63 mmol) was added, and the reaction was carried out at room temperature for 12 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography [petroleum ether: ethyl acetate = 10:1] to obtain 30 mg of a colorless oil, with a yield of 23%. 1 H NMR(600MHz,Chloroform-d)δ7.82(d,J=8.3Hz,2H),7.39(d,J=8.0Hz,2H),5.92(tt,J=56.1,4.6Hz,1H),4.20(t,J=6.1Hz,2H),2.49(s,3H),2.23(ttd,J=16.3,6.1,4.5Hz,2H).

[0330] (b) S-(3,3-difluoropropyl) 4-methylbenzenesulfonyl sulfate (I-22-28)

[0331] Using potassium p-toluenethiosulfonate (70 mg, 0.31 mmol) and I-21-28 (30 mg, 0.12 mmol) as raw materials, according to the synthesis method of I-22-16, 15 mg of a colorless oil was obtained, with a yield of 46%. 1 H NMR(600MHz,Chloroform-d)δ7.83(d,J=8.2Hz,2H),7.37(d,J=8.0Hz,2H),5.86(tt,J=56.0,4.0Hz,1H),3.08(t,J=7.6Hz,2H),2.47(s,3H),2.25(ttd,J=17.0,7.5,4.0Hz,2H).

[0332] (c) 2-(4-(((3,3-difluoropropyl)disulfanyl)methyl)phenyl)-4-methylpyrimidine-5-carboxylic acid (I-18-28)

[0333] Using I-17-9 (25 mg, 0.11 mmol), triethylamine (29 mg, 0.29 mmol) and I-22-28 (31 mg, 0.12 mmol) as raw materials, according to the synthesis method of I-18-9, 26 mg of a white solid was obtained, with a yield of 73%. 11H NMR (600 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.41 (d, J = 8.3 Hz, 2H), 7.53 (d, J = 8.3 Hz, 2H), 6.07 (tt, J = 56.4, 4.3 Hz, 1H), 4.08 (s, 2H), 2.80 (s, 3H), 2.62 (t, J = 7.7 Hz, 2H), 2.17 - 2.07 (m, 2H).

[0334] (d)(2 - (((4S,7S,10S)-10 - ((S)-4 - ((tert - butoxycarbonyl)amino)-2-(2-(4 - (((3,3 - difluoropropyl)disulfanyl)methyl)phenyl)-4 - methylpyrimidine - 5 - carboxamido)-N - methylbutanamido)-2 6 -(2 - ((tert - butoxycarbonyl)amino)ethoxy)-4 - ((cyanomethyl)carbamoyl)-7 - methyl - 6,9 - dioxo - 5,8 - diaza - 1,2(1,3)-dibenzocyclodecane - 1 6 -yl)oxy)ethyl) tert - butyl carbamate (I - 19 - 28)

[0335] Using I - 18 - 28 (21 mg, 0.06 mmol), I - 12 (50 mg, 0.05 mmol), N,N,N′,N′ - tetramethyl - O-(7 - azabenzotriazol - 1 - yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N - diisopropylethylamine (14 mg, 0.11 mmol) as starting materials, according to the synthesis method of I - 19 - 9, 44 mg of white solid was obtained with a yield of 64%. 11H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 14.0, 7.4 Hz, 2H), 8.82 (s, 1H), 8.66 (t, J = 5.6 Hz, 1H), 8.41 (d, J = 8.2 Hz, 2H), 8.36 (d, J = 8.8 Hz, 1H), 7.53 (d, J = 8.2 Hz, 2H), 7.19 (d, J = 7.9 Hz, 1H), 7.11 (s, 2H), 7.03 (dd, J = 8.7, 3.1 Hz, 1H), 7.01 - 6.95 (m, 1H), 6.75 (s, 1H), 6.72 - 6.60 (m, 3H), 6.36 (s, 1H), 6.06 (tt, J = 56.2, 4.0 Hz, 1H), 4.86 - 4.80 (m, 1H), 4.77 - 4.69 (m, 2H), 4.19 - 4.16 (m, 2H), 4.10 - 4.07 (m, 2H), 4.04 - 3.97 (m, 4H), 3.29 - 3.18 (m, 4H), 3.16 - 3.00 (m, 4H), 2.82 (s, 3H), 2.66 (s, 3H), 2.63 - 2.60 (m, 2H), 2.17 - 2.07 (m, 2H), 1.99 - 1.94 (m, 1H), 1.73 (s, 1H), 1.36 - 1.33 (m, 27H), 1.19 (d, J = 6.6 Hz, 3H).

[0336] (e) Compound 28

[0337] Using I-19-28 (37 mg, 0.03 mmol) and trifluoroacetic acid (49 mg, 0.43 mmol) as starting materials, according to the synthesis method of Compound 1, Compound 28 was prepared as a white solid (30 mg, yield 72%). 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.41 (d, J = 8.1 Hz, 2H), 8.38 (d, J = 8.5 Hz, 1H), 8.06 - 7.89 (m, 10H), 7.55 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 8.6 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 6.07 (tt, J = 56.4, 4.3 Hz, 1H), 5.03 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 4.10 (s, 2H), 3.17 - 2.97 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.64 - 2.62 (m, 2H), 2.15 - 2.09 (m, 3H), 2.03 - 1.98 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.75, 172.46, 171.47, 170.16, 166.25, 165.75, 163.40, 159.14 (q, J = 31.3 Hz), 156.31, 155.15, 154.14, 141.43, 136.04, 135.33, 131.39, 130.93, 130.25, 130.07, 129.82, 129.72, 128.67, 128.62, 128.57, 127.32, 118.03, 117.52 (q, J = 299.5 Hz), 116.97 (t, J = 238.3 Hz), 115.01, 114.78, 66.15, 65.95, 59.82, 51.40, 48.45, 48.26, 41.91, 38.70, 38.68, 36.43, 34.06, 33.64 (t, J = 21.3 Hz), 33.01, 29.99 (t, J = 5.6 Hz), 29.45, 27.79, 23.20, 19.84. HRMS (ESI): Anal. Calcd for C 47 H 58 F2N 11 O7S2[(M + H) + : 990.3925, found: 990.3926.

[0338] Example 29: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(((3,3,3-trifluoropropyl)disulfanyl)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 29)

[0339]

[0340] Using potassium p-toluenethiosulfonate and 1-iodo-3,3,3-trifluoropropane as starting materials, following the synthesis method of Compound 16, after four steps (synthesis route as above), Compound 29 was obtained as a white solid, 37 mg, 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.41 (d, J = 8.2 Hz, 2H), 8.38 (d, J = 9.8 Hz, 1H), 8.04 - 7.90 (m, 10H), 7.56 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 10.3 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.79 - 4.71 (m, 2H), 4.25 - 4.16 (m, 6H), 4.12 (s, 2H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.67 - 2.65 (m, 5H), 2.55 - 2.52 (m, 2H), 2.12 - 2.08 (m, 1H), 2.02 - 1.97 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.74, 172.45, 171.47, 170.16, 166.25, 165.74, 163.38, 158.99 (q, J = 31.8 Hz), 156.31, 155.15, 154.14, 141.47, 136.09, 135.33, 131.37, 130.93, 130.26, 130.13, 129.83, 129.72, 128.68, 128.62, 128.58, 127.34, 126.91 (q, J = 277.3 Hz), 118.03, 117.51 (q, J = 299.7 Hz), 115.02, 114.78, 66.15, 65.95, 59.82, 51.40, 48.45, 48.26, 41.92, 38.70, 38.68, 36.43, 34.06, 33.11 (q, J = 28.0 Hz), 33.00, 32.99, 29.50, 27.79, 23.20, 19.84. HRMS (ESI): Anal. Calcd for C 47 H 57 F3N 11 O7S2[(M + H) + : 1008.3830, found: 1008.3833.

[0341] Example 30: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(2-(methyldithio)ethyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 , 2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 30)

[0342]

[0343] (a) Ethyl 2-(4-(2-hydroxyethyl)phenyl)-4-methylpyrimidine-5-carboxylate (I-15-30)

[0344] Using 4-(hydroxyethyl)phenylboronic acid pinacol ester (372 mg, 1.50 mmol), I-3 (300 mg, 1.50 mmol), sodium carbonate (318 mg, 3.00 mmol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (110 mg, 0.15 mmol) as raw materials, according to the synthesis method of I-15-9, 277 mg of white solid was obtained with a yield of 65%. 11H NMR (600 MHz, Chloroform-d) δ 9.24 (s, 1H), 8.51 (d, J = 7.7 Hz, 2H), 7.41 (d, J = 7.8 Hz, 2H), 4.45 (q, J = 7.1 Hz, 2H), 3.94 (t, J = 6.6 Hz, 2H), 2.99 (t, J = 6.6 Hz, 2H), 2.93 (s, 3H), 1.46 (t, J = 7.1 Hz, 3H), 1.36 (s, 1H).

[0345] (b) Ethyl 2-(4-(2-(acetylthio)ethyl)phenyl)-4-methylpyrimidine-5-carboxylate (I-16-30)

[0346] Using I-15-30 (237 mg, 0.83 mmol), thioacetic acid (189 mg, 2.48 mmol), triphenylphosphine (649 mg, 2.48 mmol) and diisopropyl azodicarboxylate (500 mg, 2.48 mmol) as starting materials, according to the synthesis method of I-16-9, 282 mg of white solid was obtained, with a yield of 98%. 1 1H NMR (600 MHz, Chloroform-d) δ 9.20 (s, 1H), 8.46 (d, J = 6.8 Hz, 2H), 7.36 (d, J = 6.9 Hz, 2H), 4.45 - 4.40 (m, 2H), 3.16 (t, J = 7.1 Hz, 2H), 2.95 (t, J = 6.8 Hz, 2H), 2.90 (s, 3H), 2.34 (s, 3H), 1.43 (t, J = 6.7 Hz, 3H).

[0347] (c) 2-[4-(2-mercaptoethyl)phenyl]-4-methylpyrimidine-5-carboxylic acid (I-17-30)

[0348] Using I-16-30 (100 mg, 0.29 mmol) and sodium hydroxide (46 mg, 1.16 mmol) as starting materials, according to the synthesis method of I-17-9, 79 mg of white solid was obtained, with a yield of 99%. 1 1H NMR (600 MHz, DMSO-d6) δ 13.54 (s, 1H), 9.16 (s, 1H), 8.39 (d, J = 8.3 Hz, 2H), 7.42 (d, J = 8.3 Hz, 2H), 2.95 (t, J = 7.5 Hz, 2H), 2.83 - 2.79 (m, 5H), 2.32 (t, J = 7.7 Hz, 1H).

[0349] (d) 4-Methyl-2-(4-(2-(methyldithio)ethyl)phenyl)pyrimidine-5-carboxylic acid (I-18-30)

[0350] Using I-17-30 (30 mg, 0.11 mmol), triethylamine (33 mg, 0.33 mmol) and methyl methanesulfonate (17 mg, 0.13 mmol) as raw materials, according to the synthesis method of I-18-9, 22 mg of white solid was obtained with a yield of 63%. 1 H NMR (600 MHz, DMSO-d6) δ 9.07 (s, 1H), 8.36 (d, J = 7.9 Hz, 2H), 7.42 (d, J = 7.9 Hz, 2H), 3.08 - 3.03 (m, 4H), 2.78 (s, 3H), 2.43 (s, 3H).

[0351] (e)(2 - (((4S,7S,10S)-10 - ((S)-4 - ((tert-butoxycarbonyl)amino)-N-methyl-2-(4-methyl-2-(4-(2-(methyldisulfanyl)ethyl)phenyl)pyrimidine-5-carboxamido)butanamido)-2 6 -(2 - ((tert-butoxycarbonyl)amino)ethoxy)-4 - ((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-1 6 -yl)oxy)ethyl)carbamic acid tert-butyl ester (I-19-30)

[0352] Using I-18-30 (18 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I-19-9, 49 mg of white solid was obtained with a yield of 75%. 11H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 21.2, 7.4 Hz, 2H), 8.81 (s, 1H), 8.67 (t, J = 5.8 Hz, 1H), 8.38 (d, J = 8.0 Hz, 3H), 7.45 (d, J = 7.8 Hz, 2H), 7.20 (d, J = 8.3 Hz, 1H), 7.12 (s, 2H), 7.04 (d, J = 8.6 Hz, 1H), 7.02 - 6.98 (m, 1H), 6.75 (s, 1H), 6.72 - 6.63 (m, 3H), 6.37 (s, 1H), 4.87 - 4.81 (m, 1H), 4.77 - 4.71 (m, 2H), 4.18 (t, J = 5.0 Hz, 2H), 4.05 - 3.97 (m, 4H), 3.26 - 3.09 (m, 8H), 3.08 - 3.06 (m, 4H), 2.82 (s, 3H), 2.66 (s, 3H), 2.45 (s, 3H), 1.98 - 1.94 (m, 1H), 1.77 - 1.69 (m, 1H), 1.36 - 1.34 (m, 27H), 1.20 (d, J = 6.8 Hz, 3H).

[0353] (f) Compound 30

[0354] Using I-19-30 (43 mg, 0.04 mmol) and trifluoroacetic acid (59 mg, 0.52 mmol) as starting materials, according to the synthetic method of Compound 1, Compound 30 was prepared as a white solid (17 mg, yield 35%). 1 1H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.83 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.39 - 8.36 (m, 3H), 8.01 - 7.87 (m, 10H), 7.46 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 8.5 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.14 (m, 6H), 3.18 - 3.00 (m, 12H), 2.85 (s, 3H), 2.66 (s, 3H), 2.45 (s, 3H), 2.12 - 2.07 (m, 1H), 2.02 - 1.97 (m, 1H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.68, 172.36, 171.41, 170.09, 166.21, 165.64, 163.53, 159.08 (q, J = 31.8 Hz), 156.24, 155.08, 154.07, 143.86, 135.27, 135.08, 131.32, 130.87, 130.19, 129.75, 129.66, 129.55, 128.60, 128.55, 128.49, 127.11, 117.97, 117.44 (q, J = 298.8 Hz), 114.95, 114.71, 66.09, 65.90, 59.74, 51.31, 48.37, 48.19, 38.64, 38.62, 38.18, 36.37, 34.98, 34.00, 32.95, 29.41, 27.73, 23.16, 22.92, 19.79. HRMS (ESI): Anal. Calcd for C 46 H 58 N 11 O7S2[(M + H) + : 940.3957, found: 940.3962.

[0355] Example 31: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(2-(tert-butyldithio)ethyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-cyanomethyl-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 31)

[0356]

[0357] Using I-17-30 and I-7-6 as starting materials, according to the synthesis method of Compound 30, through three steps (synthesis route as above), Compound 31 was prepared, 32 mg of white solid 11H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 8.99 (d, J = 7.8 Hz, 1H), 8.83 (s, 1H), 8.76 (t, J = 5.5 Hz, 1H), 8.40 - 8.35 (m, 3H), 8.00 - 7.86 (m, 10H), 7.43 (d, J = 8.3 Hz, 2H), 7.24 (dd, J = 8.6, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.03 - 4.98 (m, 1H), 4.79 - 4.72 (m, 2H), 4.23 - 4.14 (m, 6H), 3.18 - 3.00 (m, 12H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.01 - 1.97 (m, 1H), 1.33 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.48, 170.16, 166.28, 165.71, 163.59, 159.13 (q, J = 31.8 Hz), 156.30, 155.15, 154.14, 143.87, 135.33, 135.17, 131.39, 130.93, 130.27, 129.83, 129.73, 129.55, 128.67, 128.62, 128.58, 127.18, 118.03, 117.45 (q, J = 298.5 Hz), 115.01, 114.78, 66.15, 65.96, 59.82, 51.40, 48.44, 48.25, 48.10, 41.04, 38.70, 38.68, 36.44, 35.07, 34.07, 33.01, 30.11, 29.46, 27.80, 23.22, 19.85. HRMS (ESI): Anal. Calcd for C 49 H 64 N 11 O7S2[(M + H) + : 982.4426, found: 982.4422.

[0358] Example 32: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(3-(methyldithio)propyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 , 2 6-Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 32)

[0359]

[0360] (a) Ethyl 2-(4-(3-hydroxypropyl)phenyl)-4-methylpyrimidine-5-carboxylate (I-15-32)

[0361] Using 4-hydroxypropylphenylboronic acid pinacol ester (250 mg, 0.99 mmol), I-3 (198 mg, 0.99 mmol), sodium carbonate (210 mg, 1.98 mmol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (72 mg, 0.10 mmol) as starting materials, according to the synthesis method of I-15-9, 237 mg of white solid was obtained with a yield of 80%. 1 H NMR (600 MHz, Chloroform-d) δ 9.20 (s, 1H), 8.44 (d, J = 8.2 Hz, 2H), 7.34 (d, J = 8.2 Hz, 2H), 4.42 (q, J = 7.1 Hz, 2H), 3.70 (t, J = 6.4 Hz, 2H), 2.89 (s, 3H), 2.80 (t, J = 8.1 Hz, 2H), 1.97 - 1.91 (m, 2H), 1.43 (t, J = 7.1 Hz, 3H), 1.24 (s, 1H).

[0362] (b) Ethyl 2-(4-(3-(acetylthio)propyl)phenyl)-4-methylpyrimidine-5-carboxylate (I-16-32)

[0363] Using I-15-32 (135 mg, 0.45 mmol), thioacetic acid (103 mg, 1.35 mmol), triphenylphosphine (354 mg, 1.35 mmol) and diisopropyl azodicarboxylate (273 mg, 1.35 mmol) as starting materials, according to the synthesis method of I-16-9, 130 mg of white solid was obtained with a yield of 81%. 1 H NMR (600 MHz, Chloroform-d) δ 9.23 (s, 1H), 8.47 (d, J = 8.0 Hz, 2H), 7.34 (d, J = 8.0 Hz, 2H), 4.45 (q, J = 7.1 Hz, 2H), 2.95 - 2.89 (m, 5H), 2.80 (t, J = 7.3 Hz, 2H), 2.37 (s, 3H), 1.97 (p, J = 7.4 Hz, 2H), 1.45 (t, J = 7.1 Hz, 3H).

[0364] (c) 2-(4-(3-Mercaptopropyl)phenyl)-4-methylpyrimidine-5-carboxylic acid (I-17-32)

[0365] Using I-16-32 (128 mg, 0.36 mmol) and sodium hydroxide (57 mg, 1.43 mmol) as starting materials, according to the synthesis method of I-17-9, 94 mg of white solid was obtained, with a yield of 91%. 1 H NMR (600 MHz, DMSO-d6) δ 13.51 (s, 1H), 9.14 (s, 1H), 8.37 (d, J = 8.2 Hz, 2H), 7.39 (d, J = 8.2 Hz, 2H), 2.80 (s, 3H), 2.78 - 2.71 (m, 4H), 2.38 (t, J = 7.8 Hz, 1H), 1.89 (p, J = 7.2 Hz, 2H).

[0366] (d) 4-Methyl-2-(4-(3-(methyldithio)propyl)phenyl)pyrimidine-5-carboxylic acid (I-18-32)

[0367] Using I-17-32 (30 mg, 0.10 mmol), triethylamine (32 mg, 0.31 mmol) and methyl methanesulfonate (16 mg, 0.13 mmol) as starting materials, according to the synthesis method of I-18-9, 21 mg of white solid was obtained, with a yield of 60%. 1 H NMR (600 MHz, DMSO-d6) δ 13.65 (s, 1H), 9.11 (s, 1H), 8.38 (d, J = 8.2 Hz, 2H), 7.40 (d, J = 8.0 Hz, 2H), 2.80 (s, 3H), 2.79 - 2.75 (m, 4H), 2.41 (s, 3H), 2.01 (p, J = 7.4 Hz, 2H).

[0368] (e) (2-(((4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-N-methyl-2-(4-methyl-2-(4-(3-(methyldithio)propyl)phenyl)pyrimidine-5-carboxamido)butanamido)-2 6 -(2-((tert-Butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-1 6 -yl)oxy)ethyl)carbamic acid tert-butyl ester (I-19-32)

[0369] Using I-18-32 (19 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as starting materials, according to the synthesis method of I-19-9, 45 mg of white solid was obtained with a yield of 68%. 1 H NMR (600 MHz, DMSO-d6) δ 8.98 (dd, J = 20.7, 7.5 Hz, 2H), 8.80 (s, 1H), 8.66 (t, J = 5.7 Hz, 1H), 8.36 (d, J = 8.0 Hz, 3H), 7.40 (d, J = 7.9 Hz, 2H), 7.19 (d, J = 8.3 Hz, 1H), 7.11 (s, 2H), 7.03 (d, J = 8.5 Hz, 1H), 7.00 - 6.96 (m, 1H), 6.75 (s, 1H), 6.71 - 6.62 (m, 3H), 6.36 (s, 1H), 4.85 - 4.80 (m, 1H), 4.77 - 4.70 (m, 2H), 4.17 (t, J = 4.9 Hz, 2H), 4.04 - 3.97 (m, 4H), 3.25 - 3.07 (m, 8H), 2.82 (s, 3H), 2.79 - 2.76 (m, 4H), 2.65 (s, 3H), 2.41 (s, 3H), 2.01 - 1.97 (m, 3H), 1.77 - 1.70 (m, 1H), 1.36 - 1.33 (m, 27H), 1.19 (d, J = 6.7 Hz, 3H).

[0370] (f) Compound 32

[0371] Using I-19-32 (40 mg, 0.03 mmol) and trifluoroacetic acid (55 mg, 0.48 mmol) as starting materials, according to the synthesis method of Compound 1, Compound 32 was obtained as a white solid, 9 mg, with a yield of 20%. 11H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.8 Hz, 1H), 8.83 (s, 1H), 8.80 (t, J = 5.5 Hz, 1H), 8.40 - 8.35 (m, 3H), 8.08 - 7.93 (m, 10H), 7.41 (d, J = 8.3 Hz, 2H), 7.24 (dd, J = 8.7, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.37 (s, 1H), 5.01 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.14 (m, 6H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.80 - 2.77 (m, 4H), 2.66 (s, 3H), 2.42 (s, 3H), 2.14 - 2.09 (m, 1H), 2.03 - 1.99 (m, 3H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.75, 172.48, 171.48, 170.16, 166.30, 165.69, 163.65, 159.10 (q, J = 32.3 Hz), 156.29, 155.15, 154.14, 145.41, 135.32, 134.86, 131.38, 130.92, 130.27, 129.83, 129.72, 129.28, 128.73, 128.67, 128.63, 127.11, 118.02, 117.50 (q, J = 298.5 Hz), 115.03, 114.80, 66.14, 65.95, 59.84, 51.42, 48.48, 48.27, 48.13, 38.69, 38.67, 36.76, 36.43, 34.01, 33.01, 30.45, 29.41, 27.79, 23.22, 23.01, 19.83. HRMS (ESI): Anal. Calcd for C 47 H 60 N 11 O7S2[(M + H) + : 954.4113, found: 954.4113.

[0372] Example 33: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(3-(tert-Butyldithio)propyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,26 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 33)

[0373]

[0374] Using I-17-32 and I-7-6 as starting materials, following the synthesis method of Compound 32, through three steps (synthesis route as above), Compound 33 was prepared, 28 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.9 Hz, 1H), 8.83 (s, 1H), 8.79 (t, J = 5.7 Hz, 1H), 8.40 - 8.35 (m, 3H), 8.05 - 7.92 (m, 10H), 7.40 (d, J = 8.2 Hz, 2H), 7.24 (dd, J = 8.7, 2.3 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.74 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.79 - 4.72 (m, 2H), 4.25 - 4.16 (m, 6H), 3.18 - 2.99 (m, 8H), 2.85 (s, 3H), 2.78 - 2.75 (m, 4H), 2.66 (s, 3H), 2.13 - 2.09 (m, 1H), 2.02 - 1.96 (m, 3H), 1.29 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.68, 172.39, 171.42, 170.10, 166.23, 165.62, 163.58, 159.03 (q, J = 32.3 Hz), 156.23, 155.09, 154.08, 145.27, 135.26, 134.78, 131.32, 130.87, 130.21, 129.76, 129.67, 129.23, 128.61, 128.56, 128.53, 127.05, 117.96, 117.37 (q, J = 298.3 Hz), 114.96, 114.73, 66.08, 65.89, 59.76, 51.35, 48.40, 48.20, 48.00, 38.63, 38.61, 36.37, 34.05, 33.95, 32.95, 32.04, 30.50, 29.99, 29.38, 27.73, 23.16, 19.78. HRMS (ESI): Anal. Calcd for C 50 H 66 N 11 O7S2[(M + H) + : 996.4583, found: 996.4582.

[0375] Example 34: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-((R)-1-(methyldithio)ethyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 34)

[0376]

[0377] (a) (S)-1-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)ethan-1-ol (I-14-34)

[0378] At room temperature, (S)-1-(4-bromophenyl)ethan-1-ol (1.00 g, 5.00 mmol), bis(pinacolato)diboron (2.54 g, 10.00 mmol) and potassium acetate (1.47 g, 15 mmol) were added to 20 ml of 1,4-dioxane. The mixture was stirred to dissolve, and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium(II) (366 mg, 0.50 mmol) was added. Under argon protection, the reaction was heated to 90 °C for 5 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Column chromatography [petroleum ether:ethyl acetate = 10:1] gave 1.05 g of a white solid with a yield of 85%. 1 H NMR(600MHz,Chloroform-d)δ7.80(d,J=7.6Hz,2H),7.38(d,J=7.6Hz,2H),4.92(q,J=6.6Hz,1H),1.49(d,J=6.4Hz,3H),1.34(s,12H),1.25(d,J=7.4Hz,1H).

[0379] (b) Ethyl (S)-2-(4-(1-hydroxyethyl)phenyl)-4-methylpyrimidine-5-carboxylate (I-15-34)

[0380] Using I-14-34 (248 mg, 1.00 mmol), I-3 (200 mg, 1.00 mmol), sodium carbonate (212 mg, 2.00 mmol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium(II) (73 mg, 0.10 mmol) as starting materials, following the synthetic method of I-15-9, 181 mg of a white solid was obtained with a yield of 63%. 1 H NMR(600MHz,Chloroform-d)δ9.23(s,1H),8.53(d,J=8.0Hz,2H),7.54(d,J=7.9Hz,2H),5.02(q,J=6.6Hz,1H),4.45(q,J=7.2Hz,2H),2.92(s,3H),1.56(d,J=6.5Hz,3H),1.46(t,J=7.2Hz,3H).

[0381] (c) Ethyl (R)-2-(4-(1-(acetylthio)ethyl)phenyl)-4-methylpyrimidine-5-carboxylate (I-16-34)

[0382] Using I-15-34 (100 mg, 0.35 mmol), thioacetic acid (80 mg, 1.05 mmol), triphenylphosphine (275 mg, 1.05 mmol) and diisopropyl azodicarboxylate (212 mg, 1.05 mmol) as raw materials, according to the synthesis method of I-16-9, 88 mg of white solid was obtained, with a yield of 73%. 1 H NMR (600 MHz, Chloroform-d) δ 9.24 (s, 1H), 8.51 (d, J = 8.3 Hz, 2H), 7.50 (d, J = 8.3 Hz, 2H), 4.83 (q, J = 7.2 Hz, 1H), 4.45 (q, J = 7.2 Hz, 2H), 2.93 (s, 3H), 2.33 (s, 3H), 1.71 (d, J = 7.2 Hz, 3H), 1.45 (t, J = 7.1 Hz, 3H).

[0383] (d) (R)-2-(4-(1-Mercaptoethyl)phenyl)-4-methylpyrimidine-5-carboxylic acid (I-17-34)

[0384] Using I-16-34 (60 mg, 0.17 mmol) and sodium hydroxide (28 mg, 0.70 mmol) as raw materials, according to the synthesis method of I-17-9, 47 mg of white solid was obtained, with a yield of 98%. 1 H NMR (600 MHz, DMSO-d6) δ 13.54 (s, 1H), 9.15 (s, 1H), 8.40 (d, J = 8.3 Hz, 2H), 7.57 (d, J = 8.4 Hz, 2H), 4.31 (p, J = 6.8 Hz, 1H), 3.18 (d, J = 6.1 Hz, 1H), 2.81 (s, 3H), 1.64 (d, J = 6.9 Hz, 3H).

[0385] (e) (R)-4-Methyl-2-(4-(1-(methyldithio)ethyl)phenyl)pyrimidine-5-carboxylic acid (I-18-34)

[0386] Using I-17-34 (25 mg, 0.09 mmol), triethylamine (28 mg, 0.27 mmol) and methyl methanesulfonate (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I-18-9, 21 mg of white solid was obtained, with a yield of 72%. 1 H NMR (600 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.41 (d, J = 8.1 Hz, 2H), 7.56 (d, J = 8.1 Hz, 2H), 4.35 (q, J = 7.0 Hz, 1H), 2.80 (s, 3H), 2.14 (s, 3H), 1.67 (d, J = 7.0 Hz, 3H).

[0387] (f) (2 - (((4S,7S,10S)-10 - ((S)-4 - ((tert - butoxycarbonyl)amino)-N - methyl - 2 - (4 - methyl - 2 - (4 - ((R)-1 - (methyldithio)ethyl)phenyl)pyrimidine - 5 - carboxamido)butanamido)-2 6 -(2 - ((tert - butoxycarbonyl)amino)ethoxy)-4 - ((cyanomethyl)carbamoyl)-7 - methyl - 6,9 - dioxo - 5,8 - diaza - 1,2(1,3)-dibenzocyclodecane - 1 6 -yl)oxy)ethyl) tert - butyl carbamate (I - 19 - 34)

[0388] Using I - 18 - 34 (18 mg, 0.06 mmol), I - 12 (50 mg, 0.05 mmol), N,N,N′,N′ - tetramethyl - O - (7 - azabenzotriazol - 1 - yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N - diisopropylethylamine (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I - 19 - 9, 55 mg of white solid was obtained with a yield of 84%. 1 H NMR (600 MHz, DMSO - d6) δ 8.99 (dd, J = 17.1, 7.4 Hz, 2H), 8.82 (s, 1H), 8.66 (t, J = 5.6 Hz, 1H), 8.41 (d, J = 8.1 Hz, 2H), 8.37 (d, J = 9.1 Hz, 1H), 7.57 (d, J = 8.1 Hz, 2H), 7.19 (d, J = 7.7 Hz, 1H), 7.11 (s, 2H), 7.03 (d, J = 8.6 Hz, 1H), 7.01 - 6.97 (m, 1H), 6.74 (s, 1H), 6.72 - 6.62 (m, 3H), 6.36 (s, 1H), 4.85 - 4.81 (m, 1H), 4.76 - 4.70 (m, 2H), 4.36 (q, J = 6.7 Hz, 1H), 4.18 - 4.16 (m, 2H), 4.04 - 3.96 (m, 4H), 3.25 - 3.07 (m, 8H), 2.82 (s, 3H), 2.66 (s, 3H), 2.16 (s, 3H), 1.98 - 1.93 (m, 1H), 1.73 (s, 1H), 1.67 (d, J = 7.0 Hz, 3H), 1.36 - 1.33 (m, 27H), 1.19 (d, J = 6.7 Hz, 3H).

[0389] (g) Compound 34

[0390] Using I-19-34 (47 mg, 0.04 mmol) and trifluoroacetic acid (65 mg, 0.57 mmol) as starting materials, according to the synthesis method of compound 1, compound 34 was prepared as a white solid (30 mg, yield 56%). 1 H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.41 (d, J = 8.3 Hz, 2H), 8.38 (d, J = 9.3 Hz, 1H), 8.02 - 7.89 (m, 10H), 7.58 (d, J = 8.4 Hz, 2H), 7.24 (dd, J = 8.7, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.78 - 4.71 (m, 2H), 4.37 (q, J = 6.9 Hz, 1H), 4.25 - 4.15 (m, 6H), 3.19 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.18 (s, 3H), 2.12 - 2.07 (m, 1H), 2.01 - 1.95 (m, 1H), 1.68 (d, J = 7.0 Hz, 3H), 1.22 (d, J = 6.7 Hz, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 172.75, 172.46, 171.47, 170.16, 166.25, 165.76, 163.38, 159.19 (q, J = 32.2 Hz), 156.31, 155.15, 154.14, 145.93, 136.17, 135.33, 131.39, 130.93, 130.27, 129.82, 129.72, 128.68, 128.63, 128.54, 128.52, 127.33, 118.03, 117.43 (q, J = 297.4 Hz), 115.01, 114.78, 66.16, 65.96, 59.82, 51.40, 48.52, 48.46, 48.27, 38.71, 38.68, 36.43, 34.07, 33.01, 29.46, 27.80, 23.32, 23.21, 20.48, 19.85. HRMS (ESI): Anal. Calcd for C 46 H 58 N 11 O7S2[(M + H) +:940.3957, found:940.3956.

[0391] Example 35: (4S,7S,10S)-10-((S)-4-amino-2-(2-(4-((R)-1-(tert-butyldisulfanyl)ethyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 35)

[0392]

[0393] Using I-17-34 and I-7-6 as starting materials, according to the synthesis method of Compound 34, after three steps (synthesis route as above), Compound 35 was obtained as a white solid, 33 mg, 1 1H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.6 Hz, 1H), 8.41 (d, J = 8.3 Hz, 2H), 8.38 (d, J = 8.5 Hz, 1H), 8.04 - 7.91 (m, 10H), 7.54 (d, J = 8.4 Hz, 2H), 7.24 (dd, J = 8.7, 2.3 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.78 - 4.70 (m, 2H), 4.23 - 4.15 (m, 7H), 3.18 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.09 (m, 1H), 2.03 - 1.99 (m, 1H), 1.67 (d, J = 7.0 Hz, 3H), 1.30 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (201 MHz, DMSO-d6) δ 172.78, 172.48, 171.50, 170.19, 166.28, 165.78, 163.40, 159.22 (q, J = 32.0 Hz), 156.34, 155.18, 154.17, 145.90, 136.24, 135.35, 131.41, 130.95, 130.30, 129.94, 129.84, 129.75, 128.81, 128.67, 128.55, 127.35, 118.04, 117.53 (q, J = 298.6 Hz), 115.04, 114.80, 66.17, 65.97, 59.84, 51.42, 50.41, 48.47, 48.29, 48.10, 38.71, 38.69, 36.44, 34.07, 33.02, 30.20, 29.45, 27.80, 23.22, 21.07, 19.85. HRMS (ESI): Anal. Calcd for C 49 H 64 N 11 O7S2[(M + H) + : 982.4426, found: 982.4430.

[0394] Example 36: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-((S)-1-(methyldithio)ethyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclotetradecane-4-carboxamide trifluoroacetate (Compound 36)

[0395]

[0396] Using (R)-1-(4-bromophenyl)ethan-1-ol as the starting material, following the synthetic method of Compound 34, through seven steps (the synthetic route is as above), Compound 36 was prepared, 27 mg of white solid, 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.5 Hz, 1H), 9.01 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.79 (t, J = 5.6 Hz, 1H), 8.41 (d, J = 8.3 Hz, 2H), 8.38 (d, J = 8.8 Hz, 1H), 8.06 - 7.93 (m, 10H), 7.58 (d, J = 8.4 Hz, 2H), 7.24 (dd, J = 8.7, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.72 (m, 2H), 4.37 (q, J = 6.9 Hz, 1H), 4.24 - 4.15 (m, 6H), 3.19 - 2.99 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.18 (s, 3H), 2.13 - 2.09 (m, 1H), 2.03 - 1.99 (m, 1H), 1.68 (d, J = 7.0 Hz, 3H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (201 MHz, DMSO-d6) δ 172.76, 172.48, 171.48, 170.18, 166.26, 165.78, 163.39, 159.10 (d, J = 31.4 Hz), 156.34, 155.17, 154.16, 145.94, 136.19, 135.34, 131.40, 130.95, 130.28, 129.84, 129.73, 128.69, 128.65, 128.57, 128.55, 127.34, 118.04, 117.40 (q, J = 298.8 Hz), 115.05, 114.81, 66.15, 65.96, 59.84, 51.41, 48.53, 48.48, 48.27, 38.70, 38.67, 36.43, 34.06, 33.02, 29.43, 27.80, 23.32, 23.22, 20.48, 19.84. HRMS (ESI): Anal. Calcd for C 46 H 58 N 11 O7S2 [(M + H) + : 940.3957, found: 940.3956.

[0397] Example 37: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((S)-1-(tert-butyldithio)ethyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 37)

[0398]

[0399] Using I-17-36 and I-7-6 as starting materials, following the synthetic method of Compound 34, through three steps (synthetic route as above), Compound 37 was prepared, 39 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.6 Hz, 1H), 8.41 (d, J = 8.3 Hz, 2H), 8.38 (d, J = 8.0 Hz, 1H), 8.04 - 7.92 (m, 10H), 7.54 (d, J = 8.4 Hz, 2H), 7.24 (dd, J = 8.6, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.78 - 4.71 (m, 2H), 4.23 - 4.16 (m, 7H), 3.17 - 2.98 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.09 (m, 1H), 2.03 - 1.99 (m, 1H), 1.67 (d, J = 7.0 Hz, 3H), 1.30 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR(201MHz, DMSO-d6) δ 172.77, 172.48, 171.49, 170.18, 166.27, 165.78, 163.39, 159.10 (q, J = 31.7, 31.0 Hz), 156.34, 155.17, 154.16, 145.89, 136.24, 135.35, 131.40, 130.95, 130.29, 129.84, 129.74, 128.81, 128.70, 128.67, 128.55, 127.35, 118.04, 117.49 (q, J = 298.3 Hz), 115.04, 114.81, 66.16, 65.96, 59.83, 51.41, 50.40, 48.47, 48.30, 48.27, 38.70, 38.68, 36.44, 34.06, 33.02, 30.20, 29.44, 27.80, 23.22, 21.08, 19.84. HRMS(ESI): Anal. Calcd for C 49 H 64 N 11 O7S2[(M + H) + : 982.4426, found: 982.4430.

[0400] Example 38: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(2-(methyldithio)propan-2-yl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 38)

[0401]

[0402] (a) 2-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propan-2-ol (I-14-38)

[0403] Using the starting materials 2-(4-bromophenyl)-2-propanol (2.00 g, 9.35 mmol), bis(pinacolato)diboron (2.85 g, 11.22 mmol), potassium acetate (2.75 g, 28.05 mmol) and 1,1'-bis(diphenylphosphino)ferrocene dichloropalladium (683 mg, 0.94 mmol), according to the synthesis method of I-14-34, 2.37 g of a white solid was obtained, with a yield of 97%.1 1H NMR (400 MHz, Chloroform-d) δ 7.80 (d, J = 8.3 Hz, 2H), 7.50 (d, J = 8.3 Hz, 2H), 1.84 (s, 1H), 1.58 (s, 6H), 1.34 (s, 12H).

[0404] (b) Ethyl 2-(4-(2-hydroxypropan-2-yl)phenyl)-4-methylpyrimidine-5-carboxylate (I-15-38)

[0405] Using I-14-38 (655 mg, 2.50 mmol), I-3 (500 mg, 2.50 mmol), sodium carbonate (530 mg, 5.00 mmol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (II) (183 mg, 0.25 mmol) as starting materials, according to the synthesis method of I-15-9, 394 mg of white solid was obtained with a yield of 53%. 1 1H NMR (400 MHz, Chloroform-d) δ 9.19 (s, 1H), 8.47 (d, J = 8.7 Hz, 2H), 7.61 (d, J = 8.7 Hz, 2H), 4.42 (q, J = 7.1 Hz, 2H), 2.88 (s, 3H), 1.91 (s, 1H), 1.62 (s, 6H), 1.42 (t, J = 7.1 Hz, 3H).

[0406] (c) Ethyl 2-(4-(2-(acetylthio)propan-2-yl)phenyl)-4-methylpyrimidine-5-carboxylate (I-16-38)

[0407] At room temperature, intermediate I-15-38 (300 mg, 1.00 mmol) and thioacetic acid (228 mg, 3.00 mmol) were added to 5 ml of dichloromethane, stirred to dissolve, zinc iodide (319 mg, 1.00 mmol) was added, and the reaction was carried out at room temperature for 24 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography [petroleum ether:ethyl acetate = 10:1] to obtain 241 mg of white solid with a yield of 67%. 1 1H NMR (400 MHz, Chloroform-d) δ 9.19 (s, 1H), 8.45 (d, J = 8.7 Hz, 2H), 7.66 (d, J = 8.7 Hz, 2H), 4.42 (q, J = 7.1 Hz, 2H), 2.88 (s, 3H), 2.18 (s, 3H), 1.87 (s, 6H), 1.43 (t, J = 7.1 Hz, 3H).

[0408] (d) 2-(4-(2-Mercaptopropan-2-yl)phenyl)-4-methylpyrimidine-5-carboxylic acid (I-17-38)

[0409] Using I-16-38 (182 mg, 0.51 mmol) and sodium hydroxide (81 mg, 2.03 mmol) as starting materials, according to the synthesis method of I-17-9, 135 mg of white solid was obtained, with a yield of 92%. 1 H NMR (400 MHz, DMSO-d6) δ 13.55 (s, 1H), 9.16 (s, 1H), 8.39 (d, J = 8.5 Hz, 2H), 7.73 (d, J = 8.6 Hz, 2H), 3.46 (s, 1H), 2.81 (s, 3H), 1.80 (s, 6H).

[0410] (e) 4-Methyl-2-(4-(2-(methyldithio)propan-2-yl)phenyl)pyrimidine-5-carboxylic acid (I-18-38)

[0411] Using I-17-38 (55 mg, 0.19 mmol), triethylamine (58 mg, 0.57 mmol) and methyl methanesulfonate (29 mg, 0.23 mmol) as starting materials, according to the synthesis method of I-18-9, 61 mg of white solid was obtained, with a yield of 96%. 1 H NMR (600 MHz, DMSO-d6) δ 9.14 (s, 1H), 8.41 (d, J = 8.3 Hz, 2H), 7.70 (d, J = 8.3 Hz, 2H), 2.81 (s, 3H), 2.02 (s, 3H), 1.77 (s, 6H).

[0412] (f) (2-(((4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-N-methyl-2-(4-methyl-2-(4-(2-(methyldithio)propan-2-yl)phenyl)pyrimidine-5-carboxamido)butanamido)-2 6 -(2-((tert-Butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-1 6 -yl)oxy)ethyl)carbamic acid tert-butyl ester (I-19-38)

[0413] Using I-18-38 (19 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I-19-9, 44 mg of white solid was obtained, with a yield of 66%. 1 H NMR (600 MHz, DMSO-d6) δ 8.99 (dd, J = 17.9, 7.4 Hz, 2H), 8.83 (s, 1H), 8.67 (t, J = 5.7 Hz, 1H), 8.42 (d, J = 8.4 Hz, 2H), 8.37 (d, J = 9.1 Hz, 1H), 7.72 (d, J = 8.5 Hz, 2H), 7.20 (d, J = 8.4 Hz, 1H), 7.12 (s, 2H), 7.04 (d, J = 8.6 Hz, 1H), 7.01 - 6.97 (m, 1H), 6.76 (s, 1H), 6.72 - 6.62 (m, 3H), 6.37 (s, 1H), 4.87 - 4.82 (m, 1H), 4.77 - 4.69 (m, 2H), 4.18 (t, J = 4.9 Hz, 2H), 4.05 - 3.99 (m, 4H), 3.25 - 3.08 (m, 8H), 2.83 (s, 3H), 2.67 (s, 3H), 2.06 (s, 3H), 1.99 - 1.94 (m, 1H), 1.78 (s, 6H), 1.76 - 1.70 (m, 1H), 1.37 - 1.34 (m, 27H), 1.20 (d, J = 6.7 Hz, 3H).

[0414] (g) Compound 38

[0415] Using I-19-38 (38 mg, 0.03 mmol) and trifluoroacetic acid (51 mg, 0.45 mmol) as raw materials, according to the synthesis method of Compound 1, Compound 38 was obtained, 33 mg of white solid, with a yield of 78%. 11H NMR(600MHz,DMSO-d6)δ9.18(d,J=7.0Hz,1H),8.99(d,J=7.7Hz,1H),8.84(s,1H),8.78 - 8.75(m,1H),8.41(d,J=8.0Hz,2H),8.38(d,J=9.3Hz,1H),8.01 - 7.88(m,10H),7.72(d,J=8.2Hz,2H),7.24(d,J=8.2Hz,1H),7.19(s,2H),7.10(d,J=8.4Hz,1H),6.77 - 6.72(m,2H),6.38(s,1H),5.00(q,J=7.3Hz,1H),4.80 - 4.71(m,2H),4.19(t,J=6.3Hz,6H),3.20 - 2.99(m,8H),2.85(s,3H),2.66(s,3H),2.13 - 2.09(m,1H),2.07(s,3H),2.00 - 1.96(m,1H),1.79(s,6H),1.22(d,J=6.5Hz,3H). 13 13C NMR(126MHz,DMSO-d6)δ172.23,171.94,170.96,169.65,165.73,165.25,162.79,158.48(q,J=32.4,31.9Hz),155.82,154.65,153.64,148.07,135.24,134.75,130.44,129.76,129.23,128.39,128.17,128.12,127.75,127.44,126.97,126.87,117.54,116.87(q,J=297.8Hz),114.54,114.29,65.65,65.45,59.31,52.41,50.88,47.93,47.75,38.20,38.17,38.17,35.92,32.51,28.96,28.04,27.29,23.51,22.72,19.36.HRMS(ESI):Anal.Calcd for C 47 H 60 N 11 O7S2[(M + H) + :954.4113,found:954.4110.

[0416] Example 39: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-(2-(tert-Butyldithio)propan-2-yl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-16 , 2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 39)

[0417]

[0418] Using I-17-38 and I-7-6 as starting materials, following the synthetic method of Compound 38, through three steps (synthetic route as above), Compound 39 was prepared, 34 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.5 Hz, 1H), 9.00 (d, J = 7.9 Hz, 1H), 8.84 (s, 1H), 8.77 (t, J = 5.6 Hz, 1H), 8.42 - 8.37 (m, 3H), 8.03 - 7.90 (m, 10H), 7.71 (d, J = 8.5 Hz, 2H), 7.24 (dd, J = 8.6, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.72 (m, 2H), 4.24 - 4.15 (m, 6H), 3.17 - 2.97 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.13 - 2.08 (m, 1H), 2.02 - 1.97 (m, 1H), 1.74 (s, 6H), 1.22 (d, J = 6.7 Hz, 3H), 1.11 (s, 9H). 1313C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.46, 170.16, 166.25, 165.73, 163.30, 159.08 (q, J = 32.3 Hz), 156.31, 155.15, 154.14, 149.22, 135.59, 135.33, 131.39, 130.93, 130.27, 129.82, 129.72, 128.68, 128.62, 128.23, 127.51, 127.35, 118.03, 117.34 (q, J = 298.7 Hz), 115.02, 114.78, 66.15, 65.96, 59.82, 51.69, 51.39, 48.45, 48.26, 47.28, 38.70, 38.68, 36.43, 34.06, 33.01, 30.54, 29.45, 29.27, 27.79, 23.21, 19.85. HRMS (ESI): Anal. Calcd for C 50 H 66 N 11 O7S2[(M + H) + : 996.4583, found: 996.4581.

[0419] Example 40: (4S, 7S, 10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(1-(methyldithio)cyclobutyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 40)

[0420]

[0421] Using 1-(4-bromophenyl)cyclobutanol as the starting material, according to the synthetic method of Compound 38, through seven steps (the synthetic route is as above), Compound 40 was prepared, 29 mg of white solid, 11H NMR (600 MHz, DMSO-d6) δ 9.18 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.8 Hz, 1H), 8.84 (s, 1H), 8.78 (t, J = 5.6 Hz, 1H), 8.42 (d, J = 8.2 Hz, 2H), 8.38 (d, J = 9.0 Hz, 1H), 8.04 - 7.91 (m, 10H), 7.46 (d, J = 8.3 Hz, 2H), 7.24 (dd, J = 8.7, 2.4 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.5 Hz, 1H), 6.77 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.70 (m, 2H), 4.24 - 4.14 (m, 6H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.72 - 2.68 (m, 2H), 2.68 - 2.62 (m, 5H), 2.27 - 2.22 (m, 1H), 2.12 - 2.07 (m, 1H), 2.01 - 1.97 (m, 1H), 1.92 (s, 3H), 1.90 - 1.86 (m, 1H), 1.22 (d, J = 6.6 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.68, 172.39, 171.41, 170.10, 166.20, 165.70, 163.33, 159.04 (q, J = 31.7 Hz), 156.24, 155.08, 154.07, 149.63, 135.38, 135.27, 131.32, 130.86, 130.21, 129.77, 129.63, 128.74, 128.60, 128.56, 128.26, 127.23, 126.78, 117.96, 117.44 (q, J = 298.4 Hz), 114.96, 114.72, 66.07, 65.88, 59.78, 57.54, 51.34, 48.41, 48.21, 38.65, 38.61, 36.37, 34.06, 32.95, 29.36, 27.73, 23.27, 23.14, 19.77, 16.18. HRMS (ESI): Anal. Calcd for C 48 H 60 N 11 O7S2[(M + H) + : 966.4113, found: 966.4114.

[0422] Example 41: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldithio)methyl)phenyl)-4,6-dimethylpyrimidin-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 41)

[0423]

[0424] (a) Ethyl 2-[4-(hydroxymethyl)phenyl]-4,6-dimethylpyrimidine-5-carboxylate (I-25-41)

[0425] Using 4-(hydroxymethyl)phenylboronic acid pinacol ester (273 mg, 1.17 mmol), ethyl 2-chloro-4,6-dimethylpyrimidine-5-carboxylate (250 mg, 1.17 mmol), sodium carbonate (248 mg, 2.34 mmol) and 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (85 mg, 0.12 mmol) as raw materials, according to the synthesis method of I-15-9, 313 mg of white solid was obtained, with a yield of 94%, 1 1H NMR (600 MHz, Chloroform-d) δ 8.46 (d, J = 7.9 Hz, 2H), 7.47 (d, J = 7.9 Hz, 2H), 4.77 (s, 2H), 4.45 (q, J = 7.1 Hz, 2H), 2.62 (s, 6H), 1.42 (t, J = 7.1 Hz, 3H).

[0426] (b) Ethyl 2-[4-((acetylthio)methyl)phenyl]-4,6-dimethylpyrimidine-5-carboxylate (I-26-41)

[0427] Using I-25-41 (310 mg, 1.08 mmol), thioacetic acid (247 mg, 3.25 mmol), triphenylphosphine (851 mg, 3.25 mmol) and diisopropyl azodicarboxylate (656 mg, 3.25 mmol) as raw materials, according to the synthesis method of I-16-9, 322 mg of white solid was obtained, with a yield of 87%, 1 1H NMR (600 MHz, Chloroform-d) δ 8.39 (d, J = 8.1 Hz, 2H), 7.39 (d, J = 8.0 Hz, 2H), 4.44 (q, J = 7.1 Hz, 2H), 4.17 (s, 2H), 2.61 (s, 6H), 2.36 (s, 3H), 1.42 (t, J = 7.1 Hz, 3H).

[0428] (c) 2-[4-(Mercaptomethyl)phenyl]-4,6-dimethylpyrimidine-5-carboxylic acid (I-27-41)

[0429] Using I-26-41 (50 mg, 0.15 mmol) and sodium hydroxide (29 mg, 0.73 mmol) as starting materials, according to the synthesis method of I-17-9, 38 mg of white solid was obtained, with a yield of 96%. 1 H NMR (600 MHz, DMSO-d6) δ 13.81 (s, 1H), 8.35 (d, J = 8.3 Hz, 2H), 7.50 (d, J = 8.3 Hz, 2H), 3.81 (d, J = 7.8 Hz, 2H), 2.97 (t, J = 7.9 Hz, 1H), 2.57 (s, 6H).

[0430] (d) 2-(4-((tert-Butyldithio)methyl)phenyl)-4,6-dimethylpyrimidine-5-carboxylic acid (I-28-41)

[0431] Using I-27-41 (25 mg, 0.09 mmol), triethylamine (28 mg, 0.27 mmol) and I-7-6 (18 mg, 0.11 mmol) as starting materials, according to the synthesis method of I-18-9, 22 mg of white solid was obtained, with a yield of 67%. 1 H NMR (600 MHz, DMSO-d6) δ 8.30 (d, J = 8.2 Hz, 2H), 7.42 (d, J = 8.2 Hz, 2H), 4.03 (s, 2H), 2.43 (s, 6H), 1.31 (s, 9H).

[0432] (e) (2-(((4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-2-(2-(4-((tert-butyldithio)methyl)phenyl)-4,6-dimethylpyrimidine-5-carboxamido)-N-methylbutyramido)-2 6 -(2-((tert-Butoxycarbonyl)amino)ethoxy)-4-((cyanomethyl)carbamoyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-1 6 -yl)oxy)ethyl) tert-butyl carbamate (I-29-41)

[0433] Using I-28-41 (20 mg, 0.06 mmol), I-12 (50 mg, 0.05 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (24 mg, 0.06 mmol) and N,N-diisopropylethylamine (14 mg, 0.11 mmol) as raw materials, according to the synthesis method of I-19-9, 40 mg of white solid was obtained, with a yield of 60%. 1 H NMR (600 MHz, DMSO-d6) δ 8.99 (t, J = 6.5 Hz, 2H), 8.66 (t, J = 5.7 Hz, 1H), 8.37 (d, J = 8.2 Hz, 2H), 8.34 (d, J = 6.6 Hz, 1H), 7.47 (d, J = 8.2 Hz, 2H), 7.19 (d, J = 9.0 Hz, 1H), 7.13 - 7.01 (m, 4H), 6.75 (s, 1H), 6.72 - 6.62 (m, 3H), 6.40 (s, 1H), 4.88 - 4.83 (m, 1H), 4.76 - 4.70 (m, 2H), 4.18 - 4.16 (m, 2H), 4.06 - 3.98 (m, 6H), 3.26 - 3.05 (m, 8H), 2.85 (s, 3H), 2.53 (s, 6H), 1.97 - 1.88 (m, 1H), 1.73 - 1.64 (m, 1H), 1.36 - 1.34 (m, 27H), 1.32 (s, 9H), 1.19 (d, J = 6.7 Hz, 3H).

[0434] (f) Compound 41

[0435] Using I-29-41 (34 mg, 0.03 mmol) and trifluoroacetic acid (45 mg, 0.40 mmol) as raw materials, according to the synthesis method of Compound 1, Compound 41 was obtained, 28 mg of white solid, with a yield of 72%. 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.2 Hz, 1H), 9.01 (d, J = 7.7 Hz, 1H), 8.78 (t, J = 5.5 Hz, 1H), 8.37 (d, J = 8.1 Hz, 3H), 8.07 - 7.91 (m, 10H), 7.49 (d, J = 8.1 Hz, 2H), 7.25 (d, J = 8.5 Hz, 1H), 7.21 (d, J = 8.6 Hz, 1H), 7.15 (d, J = 8.5 Hz, 1H), 7.11 (d, J = 8.6 Hz, 1H), 6.75 (d, J = 7.3 Hz, 2H), 6.43 (s, 1H), 5.06 - 5.02 (m, 1H), 4.79 - 4.71 (m, 2H), 4.26 - 4.15 (m, 6H), 4.07 (s, 2H), 3.20 - 2.97 (m, 8H), 2.91 (s, 3H), 2.52 (s, 6H), 2.12 - 2.07 (m, 1H), 2.01 - 1.97 (m, 1H), 1.33 (s, 9H), 1.22 (d, J = 6.6 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.71, 172.45, 171.61, 170.10, 167.13, 163.57, 162.07, 159.09 (q, J = 31.6, 29.8 Hz), 155.17, 154.16, 140.89, 136.31, 135.37, 131.44, 130.96, 130.18, 130.08, 129.83, 129.72, 128.70, 128.62, 128.43, 128.28, 118.03, 117.53 (q, J = 299.8 Hz), 115.08, 114.76, 66.19, 65.94, 59.70, 51.42, 48.40, 48.29, 48.08, 44.36, 38.71, 38.69, 36.41, 34.07, 32.98, 30.17, 29.22, 27.80, 22.47, 19.82. HRMS (ESI): Anal. Calcd for C 49 H 64 N 11 O7S2[(M + H) + : 982.4426, found: 982.4429.

[0436] Example 42: (4S, 7S, 10S)-10-((S)-4-Amino-2-(6-(4-((tert-butyldithio)methyl)phenyl)-4-chloronicotinamido)-N-methylbutanamido)-1 6 ,26 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 42)

[0437]

[0438] Using 4-(hydroxymethyl)phenylboronic acid pinacol ester I-14-9 and methyl 4,6-dichloronicotinate I-24-42 as starting materials, following the synthesis method of Compound 41, through six steps (synthesis route as above), Compound 42 was prepared, 32 mg of white solid, 1 HNMR(600MHz,DMSO-d6)δ9.27(d,J=7.7Hz,1H),9.01(d,J=7.7Hz,1H),8.78(t,J=5.8Hz,1H),8.70(s,1H),8.39(d,J=8.9Hz,1H),8.23(s,1H),8.15(d,J=7.9Hz,2H),8.04-7.89(m,10H),7.49(d,J=8.0Hz,2H),7.24(d,J=8.1Hz,1H),7.22-7.17(m,2H),7.10(d,J=8.5Hz,1H),6.75(d,J=7.1Hz,2H),6.35(s,1H),5.06-5.00(m,1H),4.78-4.72(m,2H),4.24-4.15(m,6H),4.06(s,2H),3.18-2.97(m,8H),2.85(s,3H),2.14-2.08(m,1H),2.03-1.98(m,1H),1.34(s,9H),1.22(d,J=6.7Hz,3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.76, 172.45, 171.12, 170.17, 164.60, 159.10 (q, J = 32.1 Hz), 158.33, 155.16, 154.15, 149.64, 141.90, 140.28, 136.12, 135.34, 131.39, 130.94, 130.37, 130.32, 129.83, 129.72, 128.66, 127.97, 127.59, 121.35, 118.03, 117.32 (q, J = 297.6 Hz), 115.03, 114.76, 66.16, 65.98, 59.91, 51.41, 48.44, 48.35, 48.25, 44.18, 38.71, 38.69, 36.47, 34.07, 33.09, 32.10, 30.16, 29.75, 27.80, 19.86. HRMS (ESI): Anal. Calcd for C 48 H 60 ClN 10 O7S2[(M + H) + : 987.3771, found: 987.3774.

[0439] Example 43: (4S, 7S, 10S)-10-((S)-4-Amino-2-(6-(4-((tert-butyldithio)methyl)phenyl)-2-methylnicotinamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 43)

[0440]

[0441] Using 4-(hydroxymethyl)phenylboronic acid pinacol ester I-14-9 and methyl 6-chloro-2-methylnicotinate I-24-43 as starting materials, following the synthetic method of Compound 41, through six steps (synthetic route as above), Compound 43 was prepared, 39 mg of white solid, 1HNMR(600MHz, DMSO-d6) δ 9.00 (d, J = 7.3 Hz, 2H), 8.77 (t, J = 5.6 Hz, 1H), 8.38 (d, J = 8.9 Hz, 1H), 8.10 (d, J = 8.1 Hz, 2H), 8.03 - 7.90 (m, 10H), 7.88 - 7.84 (m, 2H), 7.47 (d, J = 8.2 Hz, 2H), 7.24 (dd, J = 8.6, 2.3 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.74 (m, 2H), 6.38 (s, 1H), 5.00 - 4.96 (m, 1H), 4.78 - 4.72 (m, 2H), 4.23 - 4.15 (m, 6H), 4.06 (s, 2H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.65 (s, 3H), 2.11 - 2.07 (m, 1H), 2.02 - 1.98 (m, 1H), 1.34 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 13 C NMR(126MHz, DMSO-d6) δ 172.75, 172.44, 171.66, 170.20, 168.44, 159.08 (q, J = 33.4 Hz), 156.11, 155.94, 155.14, 154.14, 139.36, 137.38, 137.13, 135.33, 131.40, 130.94, 130.26, 130.19, 130.08, 129.97, 129.80, 128.69, 128.60, 127.30, 118.03, 117.48, 117.07 (q, J = 297.6 Hz), 115.03, 114.76, 66.16, 65.96, 59.79, 51.39, 48.41, 48.33, 48.26, 44.30, 38.70, 38.69, 36.50, 34.07, 33.00, 30.17, 29.46, 27.79, 23.57, 19.85. HRMS(ESI): Anal. Calcd for C 49 H 63 N 10 O7S2[(M + H) + : 967.4317, found: 967.4320.

[0442] Example 44: (4S, 7S, 10S)-10-((S)-4-Amino-2-(6-(4-((tert-butyldithio)methyl)phenyl)-4-methylnicotinamido)-N-methylbutanamido)-1 6 , 26 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 44)

[0443]

[0444] Using 4-(hydroxymethyl)phenylboronic acid pinacol ester I-14-9 and methyl 6-chloro-4-methylnicotinate I-24-44 as starting materials, following the synthetic method of Compound 41, through six steps (synthetic route as above), Compound 44 was prepared, 35 mg of white solid, 1 HNMR(600MHz,DMSO-d6)δ9.05(d,J=7.5Hz,1H),9.00(d,J=7.8Hz,1H),8.77(t,J=5.7Hz,1H),8.63(s,1H),8.38(d,J=8.9Hz,1H),8.10(d,J=8.1Hz,2H),8.02 - 7.87(m,11H),7.48(d,J=8.1Hz,2H),7.25 - 7.18(m,3H),7.10(d,J=8.5Hz,1H),6.77 - 6.74(m,2H),6.38(s,1H),5.01 - 4.97(m,1H),4.79 - 4.72(m,2H),4.24 - 4.16(m,6H),4.06(s,2H),3.18 - 2.97(m,8H),2.85(s,3H),2.49(s,3H),2.13 - 2.08(m,1H),2.03 - 1.99(m,1H),1.34(s,9H),1.22(d,J=6.7Hz,3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.76, 172.44, 171.61, 170.20, 167.38, 159.04 (q, J = 33.5 Hz), 156.76, 155.14, 154.14, 148.24, 147.12, 139.46, 137.28, 135.33, 131.39, 130.93, 130.91, 130.26, 130.21, 129.80, 128.75, 128.67, 128.62, 127.31, 122.39, 118.03, 117.00 (q, J = 295.9 Hz), 115.01, 114.78, 66.15, 65.97, 59.81, 51.39, 48.42, 48.33, 48.25, 44.29, 38.71, 38.69, 36.52, 34.07, 33.02, 30.16, 29.52, 27.80, 19.85, 19.72. HRMS (ESI): Anal. Calcd for C 49 H 63 N 10 O7S2[(M + H) + : 967.4317, found: 967.4316.

[0445] Example 45: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldisulfanyl)methyl)-3-fluorophenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide trifluoroacetate (Compound 45)

[0446]

[0447] Using (2-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)methanol I-23-45 and I-3 as starting materials, following the synthetic method of Compound 41, through six steps (synthetic route as above), Compound 45 was prepared, 36 mg of white solid, 11H NMR (600 MHz, DMSO-d6) δ 9.22 (d, J = 7.4 Hz, 1H), 9.00 (d, J = 7.8 Hz, 1H), 8.86 (s, 1H), 8.78 (t, J = 5.6 Hz, 1H), 8.38 (d, J = 8.9 Hz, 1H), 8.25 (d, J = 7.9 Hz, 1H), 8.14 (d, J = 11.1 Hz, 1H), 8.05 - 7.89 (m, 10H), 7.60 (t, J = 7.9 Hz, 1H), 7.24 (d, J = 8.5 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.76 - 6.73 (m, 2H), 6.38 (s, 1H), 5.02 - 4.97 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 4.09 (s, 2H), 3.18 - 2.98 (m, 8H), 2.85 (s, 3H), 2.67 (s, 3H), 2.15 - 2.09 (m, 1H), 2.05 - 1.99 (m, 1H), 1.32 (s, 9H), 1.22 (d, J = 6.6 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 172.75, 172.46, 171.44, 170.15, 166.08, 165.96, 162.06, 161.16 (d, J = 265.6 Hz), 159.08 (q, J = 31.7 Hz), 156.38, 155.15, 154.14, 138.65, 138.58, 135.34, 132.79 (d, J = 3.5 Hz), 131.39, 130.93, 130.27, 129.83, 129.71, 128.65 (d, J = 5.7 Hz), 128.27 (d, J = 15.5 Hz), 127.80, 124.32, 124.30, 118.03, 117.54 (q, J = 298.7 Hz), 115.03, 114.85 (d, J = 23.3 Hz), 114.79, 66.15, 65.95, 59.83, 51.41, 48.46, 48.26, 38.69, 38.67, 37.63, 36.42, 34.06, 33.01, 30.06, 29.42, 27.79, 23.16, 19.84. HRMS (ESI): Anal. Calcd for C 48 H 61 FN 11 O7S2[(M + H) + : 986.4175, found: 986.4174.

[0448] Example 46: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldithio)methyl)-3-methylphenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide trifluoroacetate (Compound 46)

[0449]

[0450] Using 4-hydroxymethyl-3-methylphenylboronic acid I-23-46 and I-3 as starting materials, following the synthetic method of Compound 41, through six steps (synthetic route as above), Compound 46 was prepared, 31 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 9.01 (d, J = 7.9 Hz, 1H), 8.83 (s, 1H), 8.78 (t, J = 5.6 Hz, 1H), 8.38 (d, J = 9.0 Hz, 1H), 8.27 (s, 1H), 8.22 (d, J = 8.3 Hz, 1H), 8.04 - 7.90 (m, 10H), 7.43 (d, J = 8.1 Hz, 1H), 7.24 (dd, J = 8.6, 2.3 Hz, 1H), 7.20 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.74 (m, 2H), 6.38 (s, 1H), 5.02 - 4.98 (m, 1H), 4.79 - 4.71 (m, 2H), 4.24 - 4.15 (m, 6H), 4.10 (s, 2H), 3.17 - 2.96 (m, 8H), 2.85 (s, 3H), 2.66 (s, 3H), 2.50 (s, 3H), 2.13 - 2.08 (m, 1H), 2.02 - 1.97 (m, 1H), 1.33 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 1313C NMR (126 MHz, DMSO-d6) δ 172.75, 172.45, 171.47, 170.16, 166.26, 165.70, 163.43, 159.10 (q, J = 31.5 Hz), 156.26, 155.15, 154.15, 139.07, 137.45, 136.27, 135.35, 131.43, 130.94, 130.25, 129.83, 129.73, 128.68, 128.63, 127.31, 126.12, 118.03, 117.55 (q, J = 298.8 Hz), 115.03, 114.79, 66.16, 65.97, 59.82, 51.40, 51.21, 48.45, 48.39, 48.26, 42.98, 38.70, 38.68, 36.43, 34.07, 33.01, 30.16, 29.56, 29.46, 27.80, 23.21, 19.85, 19.61. HRMS (ESI): Anal. Calcd for C 49 H 64 N 11 O7S2[(M + H) + : 982.4426, found: 982.4426.

[0451] Example 47: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-(tert-butyldithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxylic acid trifluoroacetate (Compound 47)

[0452]

[0453] (a) (4S,7S,10S)-10-((S)-4-((tert-Butoxycarbonyl)amino)-2-(2-(4-(tert-butyldithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-((tert-butoxycarbonyl)amino)ethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxylic acid methyl ester (I-31-47)

[0454] Under ice bath conditions, intermediate I-8-6 (19 mg, 0.06 mmol), methyl (4S,7S,10S)-10-((S)-2-amino-4-((tert-butoxycarbonyl)amino)-N-methylbutanamido)-16,26-bis(2-((tert-butoxycarbonyl)amino)ethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxylate I-30 (prepared according to the method of patent CN111386283) (50 mg, 0.06 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (25 mg, 0.07 mmol) were added to 3 ml of N,N-dimethylformamide, stirred to dissolve, N,N-diisopropylethylamine (14 mg, 0.11 mmol) was added, and the reaction was carried out under ice bath for 10 min. The reaction was monitored by TLC until complete. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure and purified by column chromatography [dichloromethane:methanol = 20:1] to obtain 52 mg of white solid with a yield of 77%. 1 H NMR(600MHz,DMSO-d6)δ9.06(d,J=8.0Hz,1H),8.99(d,J=7.2Hz,1H),8.82(s,1H),8.44-8.39(m,3H),7.75(d,J=8.5Hz,2H),7.18(d,J=8.9Hz,1H),7.12(s,2H),7.04(d,J=8.6Hz,1H),7.01-6.96(m,1H),6.71(d,J=8.0Hz,2H),6.70-6.66(m,1H),6.65-6.61(m,1H),6.37(s,1H),4.89-4.81(m,2H),4.76-4.68(m,1H),4.05-4.02(m,2H),4.00-3.97(m,2H),3.70(s,3H),3.26-3.20(m,3H),3.16-3.00(m,5H),2.82(s,3H),2.66(s,3H),1.99-1.94(m,1H),1.74(s,1H),1.36-1.32(m,36H),1.19(d,J=6.6Hz,3H).

[0455] (b)(4S,7S,10S)-10-((S)-4-((tert-butoxycarbonyl)amino)-2-(2-(4-(tert-butyldithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6-Bis(2-((tert-butoxycarbonyl)amino)ethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxylic acid (I-32-47)

[0456] At room temperature, the intermediate I-31-47 (46 mg, 0.04 mmol) was added to 2 ml of tetrahydrofuran and stirred until dissolved. 0.3 ml of an aqueous solution of lithium hydroxide (3.8 mg, 0.16 mmol) was added, and the reaction was carried out at room temperature for 1 h. The reaction was monitored by TLC until completion. Water was added, and the pH was adjusted to 3 with 1 M aqueous HCl solution. Ethyl acetate was added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 42 mg of a white solid with a yield of 93%. 1 1H NMR (600 MHz, DMSO-d6) δ 12.79 (s, 1H), 8.99 (d, J = 7.3 Hz, 1H), 8.90 (d, J = 7.9 Hz, 1H), 8.82 (s, 1H), 8.43 - 8.38 (m, 3H), 7.75 (d, J = 8.5 Hz, 2H), 7.18 (d, J = 8.8 Hz, 1H), 7.11 (s, 2H), 7.03 (d, J = 8.5 Hz, 1H), 7.00 - 6.96 (m, 1H), 6.72 (d, J = 15.5 Hz, 2H), 6.70 - 6.61 (m, 2H), 6.37 (s, 1H), 4.86 - 4.82 (m, 1H), 4.78 - 4.70 (m, 2H), 4.06 - 4.03 (m, 2H), 4.01 - 3.97 (m, 2H), 3.29 - 3.08 (m, 8H), 2.82 (s, 3H), 2.66 (s, 3H), 1.98 - 1.94 (m, 1H), 1.76 - 1.71 (m, 1H), 1.36 - 1.32 (m, 36H), 1.18 (d, J = 6.7 Hz, 3H).

[0457] (c) Compound 47

[0458] Using I-32-47 (36 mg, 0.03 mmol) and trifluoroacetic acid (51 mg, 0.44 mmol) as starting materials, compound 47 was prepared according to the synthesis method of compound 1. 28 mg of a white solid was obtained with a yield of 68%. 11H NMR (600 MHz, DMSO-d6) δ 12.84 (s, 1H), 9.19 (d, J = 7.4 Hz, 1H), 8.88 (d, J = 8.1 Hz, 1H), 8.84 (s, 1H), 8.43 - 8.40 (m, 3H), 8.39 - 7.77 (m, 10H), 7.76 (d, J = 8.6 Hz, 2H), 7.24 (dd, J = 8.7, 2.3 Hz, 1H), 7.19 (s, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 (s, 1H), 6.72 (s, 1H), 6.37 (s, 1H), 5.02 - 4.98 (m, 1H), 4.78 - 4.70 (m, 2H), 4.25 - 4.14 (m, 4H), 3.17 - 3.00 (m, 8H), 2.84 (s, 3H), 2.66 (s, 3H), 2.12 - 2.08 (m, 1H), 2.03 - 1.98 (m, 1H), 1.33 (s, 9H), 1.20 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 173.49, 172.45, 171.43, 170.16, 166.15, 165.75, 163.02, 159.07 (q, J = 31.8 Hz), 156.28, 155.08, 154.07, 142.06, 135.20, 135.14, 131.62, 130.80, 130.14, 129.74, 129.55, 128.98, 128.61, 128.59, 127.19, 126.46, 117.46 (q, J = 298.7 Hz), 114.85, 114.69, 66.07, 65.88, 59.77, 50.58, 50.02, 48.42, 48.13, 38.64, 38.62, 36.36, 33.28, 32.97, 29.78, 29.36, 23.14, 19.72. HRMS (ESI): Anal. Calcd for C 45 H 58 N9O8S2 [(M + H) + : 916.3844, found: 916.3846.

[0459] Example 48: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldisulfanyl)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 , 2 6-Bis(2-aminoethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxylic acid trifluoroacetate (Compound 48)

[0460]

[0461] Using intermediates I-18-15 and I-30 as starting materials, following the synthetic method of Compound 47, through three steps (synthetic route as above), Compound 48 was prepared, 37 mg of white solid, 1 H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.4 Hz, 1H), 8.88 - 8.85 (m, 1H), 8.84 (s, 1H), 8.42 - 8.39 (m, 3H), 8.26 - 7.61 (m, 10H), 7.51 (d, J = 8.4 Hz, 2H), 7.23 (dd, J = 8.7, 2.3 Hz, 1H), 7.19 (s, 2H), 7.09 (d, J = 8.6 Hz, 1H), 6.77 (s, 1H), 6.71 (s, 1H), 6.37 (s, 1H), 5.02 - 4.97 (m, 1H), 4.77 - 4.69 (m, 2H), 4.25 - 4.15 (m, 4H), 4.07 (s, 2H), 3.14 - 3.00 (m, 8H), 2.84 (s, 3H), 2.66 (s, 3H), 2.12 - 2.08 (m, 1H), 2.02 - 1.98 (m, 1H), 1.33 (s, 9H), 1.20 (d, J = 6.7 Hz, 3H). 13 C NMR (126 MHz, DMSO-d6) δ 173.54, 172.42, 171.42, 170.16, 166.19, 165.70, 163.34, 159.01 (q, J = 31.5 Hz), 156.25, 155.07, 154.06, 141.31, 135.93, 135.21, 131.66, 130.84, 130.13, 129.84, 129.74, 129.55, 128.66, 128.60, 128.54, 127.24, 117.47 (q, J = 298.9 Hz), 114.83, 114.69, 66.06, 65.88, 59.76, 50.66, 48.41, 48.35, 48.14, 44.22, 38.64, 38.62, 36.35, 33.31, 32.97, 30.09, 29.36, 23.15, 19.72. HRMS (ESI): Anal. Calcd for C 46 H 60 N9O8S2[(M + H)+ :930.4001,found:930.4002.

[0462] Example 49: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-(2-(methyldithio)propan-2-yl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxylate trifluoroacetate (Compound 49)

[0463]

[0464] Using intermediates I-18-38 and I-30 as starting materials, according to the synthesis method of Compound 47, after three steps (synthesis route as above), Compound 49 was prepared, 41 mg of white solid, 1 H NMR(600MHz,DMSO-d6)δ9.17(d,J=7.4Hz,1H),8.90(d,J=8.2Hz,1H),8.83(s,1H),8.43-8.38(m,3H),8.07-7.92(m,10H),7.71(d,J=8.5Hz,2H),7.23(dd,J=8.7,2.3Hz,1H),7.18(s,2H),7.09(d,J=8.6Hz,1H),6.76(s,1H),6.71(s,1H),6.36(s,1H),5.00-4.97(m,1H),4.79-4.69(m,2H),4.24-4.14(m,4H),3.15-2.96(m,8H),2.83(s,3H),2.66(s,3H),2.13-2.08(m,1H),2.06(s,3H),2.01-1.96(m,1H),1.78(s,6H),1.19(d,J=6.7Hz,3H). 1313C NMR (126 MHz, DMSO-d6) δ 173.49, 172.55, 171.48, 170.24, 166.26, 165.76, 163.30, 159.08 (q, J = 31.7 Hz), 156.30, 155.14, 154.14, 148.57, 135.73, 135.26, 131.58, 130.80, 130.22, 129.82, 129.60, 129.11, 128.68, 128.65, 128.25, 127.46, 127.35, 117.51 (q, J = 298.1 Hz), 114.96, 114.73, 66.11, 65.90, 59.84, 52.89, 50.48, 48.48, 48.18, 38.70, 38.68, 36.42, 33.03, 29.40, 28.52, 23.99, 23.20, 19.77. HRMS (ESI): Anal. Calcd for C 45 H 58 N9O8S2 [(M + H) + : 916.3844, found: 916.3844.

[0465] Example 50: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-7-methyl-6,9-dioxo-N-(2-oxoethyl)-5,8-diaza-1,2(1,3)-dibenzocyclotetradecane-4-carboxamide hydrochloride (Compound 50)

[0466]

[0467] (a) (2-(((4S,7S,10S)-10-((S)-4-((tert-butoxycarbonyl)amino)-2-(2-(4-((tert-butyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-2 6 -(2-((tert-butoxycarbonyl)amino)ethoxy)-4-((2,2-dimethoxyethyl)aminocarbonyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-1 6 -yl)oxy)ethyl)carbamic acid tert-butyl ester (I-33-50)

[0468] Under ice bath conditions, intermediate I-32-48 (50 mg, 0.04 mmol), aminoacetaldehyde dimethyl acetal (8.6 mg, 0.08 mmol) and N,N-diisopropylethylamine (16 mg, 0.12 mmol) were added to 2 ml of dichloromethane, stirred to dissolve, and 0.5 ml of N,N-dimethylformamide solution of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (23 mg, 0.06 mmol) was added. The reaction was carried out at ice bath for 1 h. The reaction was monitored by TLC until completion. Water and ethyl acetate were added for extraction. The organic layers were combined, washed with water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography [dichloromethane:methanol = 20:1] to obtain 43 mg of white solid with a yield of 80%. 1 H NMR(600MHz,DMSO-d6)δ8.98(d,J=7.2Hz,1H),8.86-8.81(m,2H),8.40(d,J=8.1Hz,2H),8.34(d,J=8.9Hz,1H),7.92(t,J=5.8Hz,1H),7.50(d,J=7.9Hz,2H),7.16(d,J=8.2Hz,1H),7.12(s,2H),7.02(d,J=8.5Hz,1H),7.01-6.95(m,1H),6.75-6.61(m,4H),6.37(s,1H),4.84(s,1H),4.79-4.71(m,2H),4.38(t,J=5.4Hz,1H),4.08-4.06(m,2H),4.04-3.97(m,4H),3.28(s,6H),3.26-3.05(m,10H),2.82(s,3H),2.67(s,3H),1.99-1.93(m,1H),1.78-1.70(m,1H),1.37-1.34(m,27H),1.33(s,9H),1.19(d,J=6.7Hz,3H).

[0469] (b) Compound 50

[0470] At room temperature, intermediate I-33-50 (38 mg, 0.03 mmol) was added to 5 ml of hexafluoroisopropanol, stirred to dissolve, and 6M HCl aqueous solution (71 μL, 0.43 mmol) was added. The reaction was carried out at room temperature for 30 min. The reaction was monitored by UPLC until completion, concentrated to dryness to obtain 31 mg of white solid of compound 50 with a yield of 99%. 11H NMR (600 MHz, DMSO-d6) δ 9.47 (s, 1H), 9.23 (d, J = 7.3 Hz, 1H), 8.99 (d, J = 8.0 Hz, 1H), 8.87 (s, 1H), 8.45 (t, J = 5.5 Hz, 1H), 8.40 (d, J = 8.1 Hz, 3H), 8.26 - 8.08 (m, 10H), 7.51 (d, J = 8.2 Hz, 2H), 7.24 (d, J = 8.3 Hz, 1H), 7.21 (s, 2H), 7.12 (d, J = 8.8 Hz, 1H), 6.77 (d, J = 14.2 Hz, 1H), 6.36 (s, 1H), 5.17 - 5.12 (m, 1H), 5.01 - 4.96 (m, 1H), 4.85 - 4.72 (m, 2H), 4.29 - 4.19 (m, 4H), 4.07 (s, 2H), 3.99 - 3.95 (m, 1H), 3.15 - 2.96 (m, 8H), 2.85 (s, 3H), 2.67 (s, 3H), 2.18 - 2.12 (m, 1H), 2.05 - 2.02 (m, 1H), 1.33 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 200.07, 172.67, 172.57, 171.46, 170.17, 166.20, 165.79, 163.35, 156.38, 155.15, 154.09, 141.35, 136.00, 135.34, 131.44, 131.24, 130.30, 130.20, 129.87, 129.78, 129.00, 128.70, 128.60, 127.31, 115.00, 66.08, 65.92, 59.89, 51.44, 49.78, 48.64, 48.42, 48.29, 44.30, 38.62, 38.59, 36.35, 34.44, 33.13, 30.17, 29.45, 29.23, 23.32, 19.90. HRMS (ESI): Anal. Calcd for C 48 H 63 N 10 O8S2[(M + H) + : 971.4266, found: 971.4265.

[0471] Example 51: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-(tert-butyldithio)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6-Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide (Compound 51)

[0472]

[0473] At room temperature, dissolve Compound 6 (34 mg, 0.02 mmol) in 0.5 ml of water, add sodium bicarbonate (12 mg, 0.15 mmol), stir at room temperature for 30 min, a large amount of solid precipitates, filter, dry, to obtain Compound 51, 19 mg of white solid, yield 87%, 1 1H NMR (600 MHz, DMSO-d6) δ 9.10 - 8.98 (m, 2H), 8.80 (s, 1H), 8.71 (t, J = 5.5 Hz, 1H), 8.41 (d, J = 8.4 Hz, 2H), 8.34 (d, J = 8.9 Hz, 1H), 7.75 (d, J = 8.4 Hz, 2H), 7.19 (dd, J = 8.6, 2.4 Hz, 1H), 7.14 - 7.09 (m, 2H), 7.04 - 7.01 (m, 1H), 6.72 (dd, J = 8.7, 2.3 Hz, 2H), 6.37 (s, 1H), 5.02 - 4.96 (m, 1H), 4.80 - 4.71 (m, 2H), 4.19 - 4.15 (m, 2H), 3.99 - 3.89 (m, 4H), 2.85 (s, 3H), 2.82 - 2.70 (m, 8H), 2.66 (s, 3H), 1.92 - 1.85 (m, 1H), 1.81 - 1.73 (m, 1H), 1.33 (s, 9H), 1.20 (d, J = 6.7 Hz, 3H).

[0474] Example 52: (4S,7S,10S)-10-((S)-4-Amino-N-methyl-2-(4-methyl-2-(4-((propyl disulfide)methyl)phenyl)pyrimidine-5-carboxamido)butanamido)-1 6 ,2 6 -Bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide (Compound 52)

[0475]

[0476] Using Compound 11 (26 mg, 0.02 mmol) as the raw material, according to the preparation method of Compound 51, Compound 52 was obtained, 16 mg of white solid, yield 89%, 11H NMR (600 MHz, DMSO-d6) δ 9.13 - 8.97 (m, 2H), 8.85 - 8.79 (m, 1H), 8.78 - 8.70 (m, 1H), 8.41 (d, J = 8.1 Hz, 2H), 8.37 - 8.32 (m, 1H), 7.53 (d, J = 8.1 Hz, 2H), 7.19 (d, J = 7.8 Hz, 1H), 7.16 - 7.08 (m, 2H), 7.03 (d, J = 8.5 Hz, 1H), 6.76 - 6.69 (m, 2H), 6.37 (s, 1H), 5.00 (s, 1H), 4.81 - 4.71 (m, 2H), 4.21 - 4.15 (m, 2H), 4.10 - 4.04 (m, 2H), 4.01 - 3.91 (m, 4H), 3.26 - 3.04 (m, 4H), 2.86 (s, 3H), 2.83 - 2.72 (m, 4H), 2.66 (s, 3H), 2.55 - 2.53 (m, 2H), 2.07 - 1.94 (m, 1H), 1.83 - 1.73 (m, 1H), 1.62 - 1.55 (m, 2H), 1.20 (d, J = 6.6 Hz, 3H), 0.88 (t, J = 7.7 Hz, 3H).

[0477] Example 53: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((butyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecyl-4-carboxamide (Compound 53)

[0478]

[0479] Using compound 13 (23 mg, 0.02 mmol) as the starting material, according to the preparation method of compound 51, compound 53 was prepared, 15 mg of white solid, yield 93%, 11H NMR (600 MHz, DMSO-d6) δ 9.09 - 8.98 (m, 2H), 8.84 - 8.80 (m, 1H), 8.76 - 8.70 (m, 1H), 8.41 (d, J = 7.8 Hz, 2H), 8.35 (d, J = 8.5 Hz, 1H), 7.53 (d, J = 7.7 Hz, 2H), 7.19 (d, J = 7.6 Hz, 1H), 7.16 - 7.09 (m, 2H), 7.03 (d, J = 8.2 Hz, 1H), 6.73 (d, J = 11.0 Hz, 2H), 6.37 (s, 1H), 5.03 - 4.95 (m, 1H), 4.78 - 4.70 (m, 2H), 4.20 - 4.16 (m, 2H), 4.07 - 4.04 (m, 2H), 4.00 - 3.91 (m, 4H), 3.26 - 3.07 (m, 4H), 2.85 (s, 3H), 2.82 - 2.71 (m, 4H), 2.67 (s, 3H), 2.55 - 2.52 (m, 2H), 2.00 - 1.92 (m, 1H), 1.80 - 1.72 (m, 1H), 1.53 - 1.49 (m, 2H), 1.31 - 1.27 (m, 2H), 1.20 (d, J = 6.3 Hz, 3H), 0.83 (t, J = 7.2 Hz, 3H).

[0480] Example 54: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutyramido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide (Compound 54)

[0481]

[0482] Using compound 15 (31 mg, 0.02 mmol) as the starting material, according to the preparation method of compound 51, compound 54 was prepared. 20 mg of white solid was obtained, with a yield of 95%. 11H NMR (600 MHz, DMSO-d6) δ 9.11 - 8.97 (m, 2H), 8.84 - 8.79 (m, 1H), 8.74 - 8.68 (m, 1H), 8.40 (d, J = 7.9 Hz, 2H), 8.35 (d, J = 9.4 Hz, 1H), 7.51 (d, J = 7.9 Hz, 2H), 7.19 (d, J = 7.9 Hz, 1H), 7.16 - 7.09 (m, 2H), 7.03 (d, J = 8.4 Hz, 1H), 6.77 - 6.71 (m, 2H), 6.37 (s, 1H), 5.08 - 4.92 (m, 1H), 4.80 - 4.72 (m, 2H), 4.22 - 4.17 (m, 2H), 4.09 - 4.05 (m, 2H), 4.00 - 3.91 (m, 4H), 3.24 - 3.08 (m, 4H), 2.85 (s, 3H), 2.82 - 2.71 (m, 4H), 2.67 (s, 3H), 2.00 - 1.95 (m, 1H), 1.81 - 1.73 (m, 1H), 1.33 (s, 9H), 1.20 (d, J = 6.5 Hz, 3H).

[0483] Example 55: (4S,7S,10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide hydrochloride (Compound 55)

[0484]

[0485] At room temperature, I-19-15 (40 mg, 0.03 mmol) was added to 5 ml of hexafluoroisopropanol and stirred to dissolve it. A 1,4-dioxane solution of 2M HCl (225 μl, 0.45 mmol) was added, and the reaction was carried out at 35 °C for 2 h under argon protection. The reaction was monitored by UPLC until completion, concentrated to dryness, and 35 mg of Compound 55 as a white solid was obtained with a yield of 99%. 11H NMR (600 MHz, DMSO-d6) δ 9.19 (d, J = 7.5 Hz, 1H), 9.02 - 8.99 (m, 1H), 8.83 (s, 1H), 8.80 - 8.77 (m, 1H), 8.40 - 8.37 (m, 3H), 8.04 - 7.91 (m, 10H), 7.51 (d, J = 8.3 Hz, 2H), 7.22 (dd, J = 8.7, 2.3 Hz, 1H), 7.20 - 7.17 (m, 2H), 7.10 (d, J = 8.6 Hz, 1H), 6.77 - 6.71 (m, 2H), 6.38 (s, 1H), 5.03 - 4.97 (m, 1H), 4.79 - 4.69 (m, 2H), 4.25 - 4.15 (m, 6H), 4.07 (s, 2H), 3.20 - 2.99 (m, 8H), 2.83 (s, 3H), 2.64 (s, 3H), 2.16 - 2.08 (m, 1H), 2.03 - 1.97 (m, 1H), 1.31 (s, 9H), 1.21 (d, J = 6.7 Hz, 3H).

[0486] Example 56: (4S, 7S, 10S)-10-((S)-4-Amino-2-(2-(4-((tert-butyldithio)methyl)phenyl)-4-methylpyrimidine-5-carboxamido)-N-methylbutanamido)-1 6 ,2 6 -bis(2-aminoethoxy)-N-(cyanomethyl)-7-methyl-6,9-dioxo-5,8-diaza-1,2(1,3)-dibenzocyclodecane-4-carboxamide sulfate (Compound 56)

[0487]

[0488] At room temperature, Compound 54 (30 mg, 0.03 mmol) was added to 1 ml of methanol, stirred to dissolve it, concentrated sulfuric acid (6 mg, 0.06 mmol) was added, and the reaction was carried out at room temperature for 30 min. After concentration to dryness, Compound 56 was obtained as a white solid (36 mg, yield 99%). 1HNMR(600MHz, DMSO-d6) δ 9.22 (d, J = 7.5 Hz, 1H), 9.06 - 8.99 (m, 1H), 8.86 (s, 1H), 8.82 - 8.78 (m, 1H), 8.42 - 8.38 (m, 3H), 8.09 - 7.99 (m, 10H), 7.59 (d, J = 8.1 Hz, 2H), 7.23 (dd, J = 8.5, 2.3 Hz, 1H), 7.21 - 7.12 (m, 2H), 7.16 (d, J = 8.2 Hz, 1H), 6.77 - 6.79 (m, 2H), 6.35 (s, 1H), 5.12 - 4.97 (m, 1H), 4.88 - 4.74 (m, 2H), 4.28 - 4.17 (m, 6H), 4.09 (s, 2H), 3.28 - 3.04 (m, 8H), 2.86 (s, 3H), 2.66 (s, 3H), 2.18 - 2.10 (m, 1H), 2.09 - 2.02 (m, 1H), 1.33 (s, 9H), 1.22 (d, J = 6.7 Hz, 3H).

[0489] Example 57: In Vitro Antibacterial Activity of the Compounds of the Present Invention

[0490] 1. Test Strains

[0491] The tested clinical isolated strains selected for in vitro antibacterial activity screening are shown in Table 1.

[0492] Table 1 Tested Clinical Isolated Strains for In Vitro Antibacterial Activity Screening

[0493]

[0494]

[0495] a Moderate resistance to meropenem; b High resistance to meropenem

[0496] Source of strains: The above strains are all clinically isolated pathogenic bacteria collected in Sichuan and Beijing regions in recent years. They were identified by the VITEK-60 automatic microbial identifier in the collection unit and then re-identified by our laboratory using conventional methods. Each strain of bacteria was subcultured into single colonies by streaking on an agar plate before the experiment, and the bacteria freshly cultured overnight at 37°C were appropriately diluted for the experiment.

[0497] Quality control strain: Escherichia coli ATCC35218 was purchased from ATCC (American Type Culture Collection).

[0498] 2. Culture Media and Culture Conditions

[0499] Culture medium: CAMHB (produced by Becton, Dickinson and Company)

[0500] Culture conditions: Incubate at 37°C for 16 - 20 h and observe the results.

[0501] 3. In vitro antibacterial assay method

[0502] The microbroth dilution method recommended by the Clinical and Laboratory Standards Institute (CLSI) M07 - A11 of the United States National Committee for Clinical Laboratory Standards was adopted to determine the minimum inhibitory concentration (MIC) of the test drug against the test strains.

[0503] The structure of the control compound G0775 is as follows:

[0504] Weigh an appropriate amount of the test sample powder (the control drug G0775 is in the form of hydrochloride). According to the solubility of the sample, dissolve the sample with an appropriate amount of DMSO, dilute it with sterile broth to different concentrations, and then add it to the 96 - well loading trough, so that the drug concentrations in the loading trough are 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.06 mg / L in turn (the concentration test range when meropenem is used as a quality control is 16 - 0.008 mg / L), and obtain the test sample. Select several colonies from the agar plate cultured for 18 - 24 h and directly make a bacterial suspension in sterile physiological saline, and adjust the concentration of the bacterial suspension to 0.5 McFarland units. Dilute the calibrated bacterial solution with CAMHB broth to (4 - 8)×10 5 CFU / mL to obtain the inoculum. Respectively pipette 100 μL of the above - mentioned test sample solutions with different concentrations into the 1st to 12th wells of a sterile 96 - well polystyrene plate, and then add 100 μL of the above - mentioned inoculum to each well. The final concentrations of the test samples in the wells are 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.06, 0.03 mg / L in turn (the concentration test range when meropenem is used as a quality control is 8 - 0.004 mg / L), and the final inoculum concentration is (2 - 4)×10 5 CFU / mL. Additionally, set up a growth control well containing 100 μL of inoculum and 100 μL of sterile broth. After mixing the test substances and the inoculum in each well, seal them. Place the 96 - well plate after sample addition and inoculation in an incubator at (35 ± 2)°C for 16 - 20 h. After the culture is completed, observe the growth of bacteria in each well, and take the lowest drug concentration that completely inhibits the growth of bacteria in the well as its minimum inhibitory concentration (MIC).

[0505] 4. In vitro antibacterial activity

[0506] Table 2 Experimental results of in vitro antibacterial activity of some compounds of the present invention

[0507]

[0508]

[0509] a Escherichia coli. b Klebsiella pneumonia. c Acinetobacterbaumannii. d Pseudomonas aeruginosa. e Indicates not measured.

[0510] As can be seen from Table 2, the representative compounds of the present invention have strong in vitro antibacterial activity, and have strong antibacterial activity against Escherichia coli that is moderately resistant to clinically isolated carbapenem (meropenem), and highly resistant Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa. In particular, Acinetobacter baumannii that is insensitive to the positive control compound G0775 and Pseudomonas aeruginosa that is almost ineffective against G0775 still maintain high activity, expanding the antibacterial spectrum of G0775 and overcoming the defects of G0775 with weak antibacterial activity and narrow antibacterial spectrum. In particular, for compounds 4, 6, 8, 12, 14, 15, 31, 32, 37 and 40, compared with G0775, the antibacterial activity against meropenem-resistant Acinetobacter baumannii is increased by 4, 2 - 8, 2 - 8, 4 - 8, 4 - 8, 8 - 16, 2 - 8, 4, 4 - 8, 4 times respectively, and the antibacterial activity against meropenem-resistant Pseudomonas aeruginosa is increased by 4 - 8, 4 - 8, 8 - 16, 4 - 16, 4 - 8, 4 - 8, 4 - 8, 4 - 8, 8 - 16, 8 times respectively.

[0511] The compounds of the present invention have novel structures and unique action mechanisms, and there is no cross-resistance with existing marketed drugs, having great development value.

[0512] Example 58: Pharmacokinetic study of the compounds of the present invention in rats

[0513] Healthy male SD rats were randomly divided into five groups, with 3 rats in each group. The test compound (G0775 in free base form) was injected intravenously (IV). The specific arrangement is shown in Table 3 below:

[0514] Table 3 Administration plan for pharmacokinetic experiments in rats:

[0515]

[0516] At 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8, 10, and 24 h after intravenous injection, 0.2 mL of whole blood was collected from the jugular vein at the above - set time points into an EP tube containing EDTA - K2, immediately placed on ice after collection, centrifuged at 11000 rpm for 5 min within 5 min, the plasma was separated, immediately frozen in a - 70 °C refrigerator, and analyzed and tested after an appropriate time. The analysis results are shown in Table 4.

[0517] Table 4 Pharmacokinetic Results of Rats after Intravenous Injection

[0518]

[0519]

[0520] The representative compounds of the present invention have excellent in - vivo metabolic properties in rats. When intravenously injected at a dose of 10 mg / kg, for Compound 51 and Compound 54, compared with G0775, the exposure (AUC) in rats increased by 2.8 - fold and 2.3 - fold respectively, the mean residence time (MRT) was prolonged by 1.7 - fold and 2.4 - fold respectively, and the half - life (T 1 / 2 ) was prolonged by 1.9 - fold and 2.8 - fold respectively.

[0521] Example 59: In - vivo Protective Effect of the Compounds of the Present Invention on the Thigh Infection Model of Mice

[0522] 1. Test Strains

[0523] The test clinical isolated strain selected for in - vivo pharmacodynamic study was Pseudomonas aeruginosa Pae T - 2022 - D2 - 048, clinically isolated from Nanfang Hospital of Southern Medical University. The sensitivity results of each drug are shown in Table 5:

[0524] Table 5 In - vitro Antibacterial Test Results of Each Drug against Pae T - 2022 - D2 - 048

[0525]

[0526] 2. Experimental Animals

[0527] Male ICR mice (SPF grade), body weight: 18 - 22 g.

[0528] 3. Establishment of Immunocompromised Mouse Model

[0529] For each experimental group of mice, cyclophosphamide at a dose of 150 mg / kg was first administered 4 days before infection, and cyclophosphamide at a dose of 100 mg / kg was administered 1 day before infection to immunosuppress the mice.

[0530] 4. Preparation of Bacterial Suspension

[0531] Inoculate Pseudomonas aeruginosa (Pae T-2022-D2-048) onto the slant of nutrient agar medium and culture it at 35 - 37°C for 18 - 24 h. Then inoculate it into 100 mL of nutrient broth liquid medium and culture it at 35 - 37°C for 18 - 24 h. After that, centrifuge at 3000 rpm / min for 5 min to collect the bacterial cells, discard the supernatant, resuspend with an equal volume of normal saline to obtain the initial bacterial suspension, and adjust it to an appropriate concentration with sterile normal saline for standby.

[0532] 5. Infection of the thigh muscle of immunocompromised mice

[0533] Inject the bacterial suspension of Pseudomonas aeruginosa (Pae T-2022-D2-048) into the thigh muscle of immunocompromised mice (except for the blank control group) for infection, injecting 0.1 mL of the bacterial suspension into each of the left and right legs.

[0534] 6. Administration of drugs

[0535] According to the experimental grouping, after 2 h of infection, the meropenem group and each dose group of the test article are administered by tail vein injection according to the dose (G0775 is in the free base form), and the administration volume for each mouse is 0.1 mL / 10 g. There are 8 mice in each group, and the grouping is shown in Table 6:

[0536] Table 6. Grouping for in vivo pharmacodynamic experiments

[0537]

[0538]

[0539] 7. Sample collection and colony counting

[0540] For the 2 h infection group, after 2 h of infection, sacrifice the mice, weigh the tissues, immediately put them into 5 mL of sterile normal saline, and perform tissue homogenization after storing in an ice box. For the remaining groups, after 24 h of thigh muscle infection, sacrifice the mice, weigh the tissues, immediately put them into 5 mL of sterile normal saline, perform tissue homogenization after storing in an ice box, and serially dilute the homogenate 10-fold with sterile normal saline. Take 1 mL of the diluted homogenate and add it to a sterile petri dish, then add CAB and mix well. Incubate in an incubator at 35 - 37°C for 36 - 48 h, and then perform colony counting.

[0541] 8. Experimental results

[0542] The results of colony counting are as Figure 1 shown. It can be seen that the representative compound of the present invention has excellent in vivo pharmacodynamic effects, significantly superior to G0775 and meropenem. From Figure 1It can be seen that compound 54 shows dose-dependent activity in inhibiting Pseudomonas aeruginosa in mice. When administered at a dose of 12 mg / kg, the CFU decreased by 3.5-Log relative to the infected control group, even lower than the pre-treatment level, indicating significant efficacy. In contrast, G0775 not only has poor efficacy but also lacks dose-dependence. Meropenem shows almost no activity when administered at a dose of 40 mg / kg.

[0543] All documents mentioned in this invention are cited herein as references, as if each document was individually cited as a reference. In addition, it should be understood that after reading the above teachings of this invention, those skilled in the art can make various changes or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. A macrocyclic lipopeptide compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof: Wherein, R1 is a hydroxyl group, an amino group, R2 and R3 are each independently H or a substituted or unsubstituted C1-C6 alkyl group; or, R2 and R3 may be connected to the adjacent carbon atom to form a 3-7 membered cycloalkane ring or a 3-7 membered heteroalkane ring; wherein, the C1-C6 alkyl group is optionally substituted by one or more of the following groups: C1-C6 alkyl group, C1-C6 alkoxy group, halogen and amino group; n is 0, 1, 2, 3, 4, 5 or 6; R4 is H, substituted or unsubstituted C1-C 10 alkyl, substituted or unsubstituted 3-8 membered cycloalkyl or substituted or unsubstituted C6-C 10 aryl, wherein the C1-C 10 alkyl, 3-8 membered cycloalkyl, C6-C 10 aryl is optionally substituted by one or more of the following groups: phenyl, C2-C6 alkenyl, C2-C6 alkynyl, halogen, C1-C6 alkoxy, C1-C6 alkylthio and C1-C6 alkylsilyl; X is a substituted or unsubstituted C6-C 10 aryl ring or a substituted or unsubstituted 3- to 8-membered heteroaryl ring, and the C6-C 10 aryl ring or 3- to 8-membered heteroaryl ring is optionally substituted by one or more of the following groups: C1-C6 alkyl, C1-C6 alkoxy, halogen, and amino; Y is a substituted or unsubstituted C6-C 10 aryl ring or a substituted or unsubstituted 3- to 8-membered heteroaryl ring, wherein the C6-C 10 aryl ring or 3- to 8-membered heteroaryl ring is optionally substituted with one or more of the following groups: C1-C6 alkyl, C1-C6 alkoxy, halogen, and amino.

2. The macrocyclic lipopeptide compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, The macrocyclic lipopeptide compound has a structure represented by the general formula (II): Wherein, R1, R2, R3, R4, X, Y, n are as described in claim 1.

3. The macrocyclic lipopeptide compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, R1 is a hydroxyl group, R2 and R3 are each independently H or a substituted or unsubstituted C1-C4 alkyl group, or R2 and R3 may be connected to the adjacent carbon atom to form a 3-4 membered cycloalkane ring or a 3-4 membered heteroalkane ring; n is 0, 1, 2, 3, 4, 5 or 6; R4 is a substituted or unsubstituted C1-C8 alkyl group, a substituted or unsubstituted 3-8 membered cycloalkyl group or a substituted or unsubstituted 6 membered aryl group, and the C1-C8 alkyl group, 3-8 membered cycloalkyl group, 6 membered aryl group are optionally substituted by one or more of the following: phenyl group, C2-C4 alkenyl group, C2-C4 alkynyl group, halogen, C1-C4 alkoxy group, C1-C4 alkylthio group and C1-C4 alkylsilyl group; X is a substituted or unsubstituted 3-8 membered nitrogen-containing aromatic heterocycle, and the 3-8 membered nitrogen-containing aromatic heterocycle is optionally substituted by one or more of the following groups: C1-C6 alkyl group, halogen and amino group; Y is a substituted or unsubstituted benzene ring, and the benzene ring is optionally substituted by one or more of the following: C1-C6 alkyl group and halogen.

4. The macrocyclic lipopeptide compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that, R1 is a hydroxyl group, R2 and R3 are each independently H or a C1-C3 alkyl group, or, R2 and R3 may be connected to the adjacent carbon atom to form cyclopropane or cyclobutane; n is 0, 1, 2, 3, 4, 5 or 6; R4 is a C1-C8 alkyl group, a 3-6 membered cycloalkyl group or a 6 membered aryl group, wherein the C1-C8 alkyl group is optionally substituted by halogen; X is a substituted or unsubstituted 5-6 membered aromatic heterocycle containing 1-2 nitrogen atoms, and the aromatic heterocycle is optionally substituted by one or more of the following groups: C1-C3 alkyl group and halogen; Y is a substituted or unsubstituted benzene ring, and the benzene ring is optionally substituted by one or more of the following: C1-C6 alkyl group and halogen.

5. The macrocyclic lipopeptide compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1 or 4, characterized in that, X is wherein "*" represents the linking site adjacent to the amide, and Ra and Rb are each independently H, halogen or C1-C4 alkyl; Y is wherein "*" represents the linking site near the disulfide bond, and Rc is H, halogen or C1-C4 alkyl.

6. The macrocyclic lipopeptide compound represented by the general formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, R1 is R2 and R3 are each independently H or a C1-C3 alkyl group, or, R2 and R3 may be connected to the adjacent carbon atom to form cyclopropane or cyclobutane; n is 0, 1, 2 or 3; R4 is a C1-C6 alkyl group, a 3-6 membered cycloalkyl group or a 6 membered aryl group, wherein the C1-C6 alkyl group is optionally substituted by halogen; X is where "*" represents the linkage site close to the amide; Y is where "*" represents the linking site close to the disulfide bond.

7. The macrocyclic lipopeptide compound represented by the general formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The macrocyclic lipopeptide compound represented by the general formula (I) is selected from the following group:

8. A pharmaceutical composition, comprising: One or more of the macrocyclic lipopeptide compounds represented by the general formula (I) as described in claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

9. Use of a macrocyclic lipopeptide compound represented by the general formula (I) as described in claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a pharmaceutical composition, characterized in that, The pharmaceutical composition is used for preventing and / or treating infectious diseases.

10. The use according to claim 9, characterized in that, The infectious diseases are infectious diseases caused by multi-drug resistant bacteria.

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