Polypeptide later-stage modification and construction method applied to polypeptide coupling medicine

Abstract

The invention relates to a polypeptide later-stage modification method and a construction method applied to a polypeptide coupling drug. Specifically, the invention provides mild polypeptide modification for specific sites by taking copper as a catalyst and phenylalanine boric acid side chains and imidazole derivatives as substrates, and the preparation method has the advantages of mild conditions, simplicity and convenience in operation, wide application range and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical synthesis, and in particular to a method for late-stage modification of a polypeptide and its application in the construction of a polypeptide-coupled drug. Background Art

[0002] Imidazole is a common key pharmacophore in medicinal chemistry, especially in small molecule drugs, and therefore may provide potential benefits when introduced into peptides, such as improved protease stability and enhanced biological activity. However, imidazole is widely used as a building block or directing group in small molecule organic chemistry, but has rarely been reported in the modification of peptides due to issues such as site specificity, compatibility, chemical selectivity, and maintaining peptide integrity.

[0003] The Cu-catalyzed Chan-Lam reaction, first discovered in 1998, is a landmark method for constructing sp2 CN bonds via oxidative CN cross-coupling. The reaction has been shown to be compatible with a variety of functional groups, including amines, amides, sulfonamides, imidazoles, tetrazoles, and phenols, demonstrating its versatility. As a result, the Chan-Lam reaction has been widely used in the development of active pharmaceutical ingredients (APIs) and has had a significant impact on the development of small molecule drugs. In 2018, Ball et al. developed a groundbreaking biorthogonal Chan-Lam coupling on a pyroglutamic acid-histidine dipeptide sequence using histidine as a directing group. More recently, Dang et al. used the Chan-Lam reaction to site-directedly introduce the corresponding group in a specially designed peptide containing histidine at a specific position. In this case, the imidazole side chain of histidine acts as an anchoring group to coordinate with Cu(II) and Asp, Asn, or Ser. These strategies require the deliberate use of "histidine-containing metal binding" tags in peptides / proteins to achieve effective conjugation. It is still uncertain whether imidazole-containing compounds can serve as a universal bridge for site-specific peptide modification. The development of site-specific peptide modification tools using imidazole as a substrate can promote the creation of new bioactive conjugates, cyclic peptides, etc. Summary of the invention

[0004] Problem that the invention aims to solve

[0005] In order to solve the above problems existing in the prior art, the present invention provides a mild coupling method using copper as a catalyst, phenylalanine boronated side chain and imidazole derivatives as substrates, which can be used for site-specific late-stage polypeptide modification. In the method of the present invention, imidazole derivatives have unique selectivity for a variety of unprotected amino acid side chains, and can be used to couple various imidazole derivatives including those with strong electron-withdrawing groups with a variety of amino acids and biologically active long peptides to obtain a variety of novel peptide-drug conjugates (PDCs), and can be used for the construction of novel cyclic peptide molecules.

[0006] Solutions for solving problems

[0007] A method for preparing a compound represented by formula (III), comprising reacting a compound represented by formula (I) and a compound represented by formula (II) in the presence of a copper catalyst to obtain a compound represented by formula (III),

[0008]

[0009] wherein R1 is selected from hydrogen, halogen, nitro, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl;

[0010] R2, R3, and R4 are each independently selected from hydrogen, halogen, nitro, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; or,

[0011] R2 and R3 are linked together with the carbon atom to which they are attached to form a substituted or unsubstituted aryl, heteroaryl or heterocyclic group;

[0012] The substitution is by at least one selected from hydrogen, deuterium, halogen, cyano, hydroxyl, amino, ester, substituted or unsubstituted alkyl, substituted or unsubstituted haloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkoxy, substituted or unsubstituted haloalkoxy, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl and substituted by a substituent;

[0013] R5 is hydrogen or an amino protecting group.

[0014] Effects of the Invention

[0015] In the method of the present invention, a Chan-Lam reaction is performed in the presence of a copper catalyst using a boronic acid derivative represented by formula (I) and an imidazole derivative represented by formula (II) as substrates to achieve site-specific modification in a polypeptide compound. In the method provided by the present invention, the imidazole derivative has a unique selectivity for a variety of unprotected amino acid side chains (excluding cysteine ​​and arginine), and can be used to couple various imidazole derivatives including those with strong electron-withdrawing groups with a variety of amino acids and biologically active long peptides, thereby obtaining a variety of novel polypeptide-drug conjugates (PDCs), and can be used for the construction of novel cyclic peptide molecules, and can accurately adjust the physicochemical properties of peptides, for example, the stability of polypeptide compounds and cell membrane permeability. The wide range of imidazole derivatives will provide unprecedented flexibility and structural diversity for exploring reactivity in a biological context.

[0016] The preparation method provided by the present invention has mild reaction conditions and high yield; the method can be applied to site-specific modification of peptide chains of different lengths and has wide applicability; the method provided by the present invention has good chemical selectivity and potential for application in the modification of polypeptides and has wide application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is the ESI spectrum of compound 52;

[0018] Figure 2 is the ESI spectrum of compound 53. DETAILED DESCRIPTION

[0019] In order to make the technical solutions and beneficial effects of the present invention more clearly understandable, the following is a detailed description by listing specific embodiments. The drawings are not necessarily drawn to scale, and local features may be enlarged or reduced to more clearly show the details of the local features; unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application belongs.

[0020] The late-stage site-specific modification of peptides plays a key role in the coupling of bioactive molecules and drug development, and there is still a need to develop chemical methods that can achieve site-specific modification of peptides under milder conditions. After extensive and in-depth research, the inventors have developed a peptide site-specific late-stage modification strategy based on the Chan-Lam coupling reaction, which uses phenylalanine-based boric acid side chains and imidazole derivatives to couple imidazole functional group-containing compounds to complex peptides using a copper catalyst to catalyze the Chan-Lam coupling reaction. Different from the prior art, the present invention utilizes the mild reaction conditions of the Chan-Lam coupling and the unique nucleophilicity of imidazole to develop a new modification strategy for peptide compounds. Based on this, the present invention is completed.

[0021] The present invention provides a method for preparing a compound represented by formula (III), wherein a compound represented by formula (I) and a compound represented by formula (II) are reacted in the presence of a copper catalyst to obtain a compound represented by formula (III).

[0022]

[0023] wherein R1 is selected from hydrogen, halogen, nitro, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl;

[0024] R2, R3, and R4 are each independently selected from hydrogen, halogen, nitro, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; or,

[0025] R2 and R3 are linked together with the carbon atom to which they are attached to form a substituted or unsubstituted aryl, heteroaryl or heterocyclic group;

[0026] The substitution is by at least one selected from hydrogen, deuterium, halogen, cyano, hydroxyl, amino, ester, substituted or unsubstituted alkyl, substituted or unsubstituted haloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkoxy, substituted or unsubstituted haloalkoxy, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl and substituted by a substituent;

[0027] R5 is hydrogen or an amino protecting group.

[0028] In certain embodiments, the copper catalyst is selected from one or more of Cu(OAc)2, CuCl, CuCl2, CuBr, CuBr2, Cu(SO3CF3)2, Cu(HCOO)2·4H2O, CuSO4, [Cu(L1)], [Cu(L2)], [Cu(L3)], [Cu(L4)], [Cu(L5)], [Cu(L6)], [Cu(L7)], and [Cu(L8)]; wherein the structural formulas of [Cu(L1)], [Cu(L2)], [Cu(L3)], [Cu(L4)], [Cu(L5)], [Cu(L6)], [Cu(L7)], and [Cu(L8)] are as follows:

[0029]

[0030] In certain embodiments, the copper catalyst is selected from one or more of [Cu(L1)], [Cu(L2)], [Cu(L3)], [Cu(L4)], [Cu(L5)], [Cu(L6)], [Cu(L7)], and [Cu(L8)].

[0031] In certain embodiments, the copper catalyst is selected from [Cu(L6)].

[0032] In certain embodiments, the method is performed in the presence of a solvent, and the solvent is selected from one or more of MeOH, DMF, DCM, MeCN, i-PrOH, EtOH, DMA and THF.

[0033] In certain embodiments, the solvent is selected from one or more of MeOH, MeCN, and EtOH.

[0034] In certain embodiments, the solvent is selected from MeOH.

[0035] In certain embodiments, the method is performed in the presence of a base, and the base is selected from an organic base or an inorganic base.

[0036] In certain embodiments, the organic base is selected from one or more of TEA, DIPEA, DABCO, TEMED, MeONa, t-BuOK, 2,6-Lutidine, and Py.

[0037] In certain embodiments, the organic base is selected from one or more of TEA, DIPEA, TEMED, and MeONa.

[0038] In certain embodiments, the organic base is selected from DIPEA.

[0039] In certain embodiments, the inorganic base is selected from one or more of NaHCO3, Na2CO3, and K3PO4.

[0040] In certain embodiments, the inorganic base is selected from NaHCO3.

[0041] In certain embodiments, the compound represented by formula (I) is a compound represented by formula (I-1):

[0042]

[0043] Wherein, R6 is selected from a peptide chain or an amino protecting group;

[0044] R7 is selected from a peptide chain, a carboxyl protecting group, a hydroxyl group and an amino group.

[0045] In certain embodiments, R6 is a peptide chain formed by connecting amino acid residues.

[0046] In certain embodiments, the amino acid is selected from one or more of substituted or unsubstituted Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro, Lys, Trp, Asp, Gly, Ser, Glu, Thr, His;

[0047] In certain embodiments, the amino acid is selected from one or more of substituted or unsubstituted Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro.

[0048] In certain embodiments, the amino acid is selected from Tyr.

[0049] The substitution is substituted with at least one substituent selected from amino or Boc.

[0050] In certain embodiments, R6 is an amino protecting group selected from Ac and / or Boc.

[0051] In certain embodiments, R7 is selected from a peptide chain formed by connecting amino acid residues.

[0052] In certain embodiments, the amino acid is selected from one or more of substituted or unsubstituted Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro, Lys, Trp, Asp, Asn, Gly, Ser, Glu, Gln, Thr, and His.

[0053] In certain embodiments, the amino acid is selected from one or more of substituted or unsubstituted Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro, Lys, Trp, Asp, Gly, Ser, Glu, Thr, His;

[0054] The substitution is substituted with at least one substituent selected from amino or Boc.

[0055] In certain embodiments, the length of the peptide chain is 1 to 50 amino acids, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, etc.

[0056] In certain embodiments, the peptide chain is 1 to 30 amino acids in length.

[0057] In certain embodiments, the length of the peptide chain is 1 to 25 amino acids.

[0058] In certain embodiments, the length of the peptide chain is 1 to 20 amino acids.

[0059] In certain embodiments, the R7 is a carboxyl protecting group, and the carboxyl protecting group is selected from methoxy.

[0060] In certain embodiments, the compound represented by formula (I-1) is any one of the following:

[0061]

[0062]

[0063] In certain embodiments, R2, R3, and R4 are each independently selected from hydrogen, halogen, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3～8 Cycloalkyl, substituted or unsubstituted 3-7 membered heterocyclic group, substituted or unsubstituted C 6～10 Aryl, substituted or unsubstituted 5- to 10-membered heteroaryl.

[0064] In certain embodiments, R2, R3, and R4 are each independently selected from hydrogen, halogen, nitro, substituted or unsubstituted C 3～8 Cycloalkyl, substituted or unsubstituted 3-7 membered heterocyclic group, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 6～10 Aryl.

[0065] In certain embodiments, R2, R3, and R4 are each independently selected from hydrogen, halogen, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 6～10 Aryl.

[0066] In certain embodiments, the halogen is selected from fluorine, chlorine, and bromine.

[0067] In certain embodiments, R2 and R3 together with the carbon atoms to which they are attached are linked to form a substituted or unsubstituted C 6-10 Aryl, 5- to 10-membered heteroaryl or 3- to 7-membered heterocyclic group.

[0068] In certain embodiments, R2 and R3 together with the carbon atoms to which they are attached are linked to form a substituted or unsubstituted C 6-10 Aryl.

[0069] The substitution is by at least one selected from hydrogen, deuterium, halogen, hydroxyl, amino, C 1～6 Alkyl, C 1～6 Haloalkyl, C 3～8 Cycloalkyl, 3-7 membered heterocyclic group, C 1～6 Alkoxy, C 1～6 Haloalkoxy, C 6～10 Aryl, 5- to 6-membered heteroaryl and substituted by a substituent.

[0070] In certain embodiments, the substitution is by at least one selected from hydrogen, deuterium, halogen, amino and substituted by a substituent.

[0071] R5 is an amino protecting group.

[0072] In certain embodiments, R5 is Boc.

[0073] Preferably, the compound represented by formula (II) is any one of the following:

[0074]

[0075] In certain embodiments, the temperature of the reaction is 10-50°C, such as 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C.

[0076] In certain embodiments, the temperature of the reaction is 20-40°C.

[0077] In certain embodiments, the reaction time of the reaction is 10-96 h, for example, 10 h, 15 h, 20 h, 25 h, 30 h, 35 h, 40 h, 45 h, 50 h, 55 h, 60 h, 65 h, 70 h, 75 h, 80 h, 85 h, 90 h, etc.

[0078] In certain embodiments, the reaction time of the reaction is 20-72 hours.

[0079] In certain embodiments, the molar ratio of the compound represented by formula (I) to the compound represented by formula (II) is 1:1 to 5, for example, 1:1, 1:2, 1:3, 1:4, 1:5.

[0080] In certain embodiments, the molar ratio of the compound represented by formula (I) to the compound represented by formula (II) is 1:1-3.

[0081] In certain embodiments, the molar ratio of the compound represented by formula (I) to the compound represented by formula (II) is 1:2.

[0082] In certain embodiments, the molar ratio of the compound represented by formula (I) to the copper catalyst is 1:0.1-1, for example, 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1.

[0083] In certain embodiments, the molar ratio of the compound represented by formula (I) to the copper catalyst is 1:0.1 to 0.5.

[0084] In certain embodiments, the molar ratio of the compound represented by formula (I) to the copper catalyst is 1:0.2.

[0085] In certain embodiments, the molar ratio of the compound represented by formula (I) to the base is 1:1 to 5, for example 1:1, 1:2, 1:3, 1:4, 1:5.

[0086] In certain embodiments, the molar ratio of the compound represented by formula (I) to the base is 1:1-3.

[0087] In certain embodiments, the molar ratio of the compound represented by formula (I) to the base is 1:2.

[0088] Term Explanation

[0089] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0090] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms, and more preferably an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof. More preferred are lower alkyl groups having 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of D atoms, halogen, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0091] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, preferably 3 to 8 (e.g., 3, 4, 5, 6, 7, and 8) carbon atoms, and more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include cycloalkyls of spirocyclic, condensed, and bridged rings.

[0092] The term "spirocycloalkyl" refers to a polycyclic group of 5 to 20 yuan, a carbon atom (called spiral atom) shared between monocyclic rings, which may contain one or more double bonds. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan (e.g., 7, 8, 9 or 10 yuan). According to the number of spiral atoms shared between rings, the spirocycloalkyl is divided into a single spiral cycloalkyl, a double spiral cycloalkyl or a multi-spirocycloalkyl, preferably a single spiral cycloalkyl and a double spiral cycloalkyl. More preferably, it is 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral cycloalkyl. Non-limiting examples of spirocycloalkyl include:

[0093]

[0094] The term "condensed cycloalkyl" refers to 5 to 20 yuan, and each ring in the system shares a pair of adjacent carbon atoms with other rings in the system. All carbon polycyclic groups, wherein one or more rings can contain one or more double bonds. Preferably 6 to 14 yuan, more preferably 7 to 10 yuan (such as 7,8,9 or 10 yuan). According to the number of the composition ring, dicyclo, tricyclo, tetracycle or polycyclic condensed cycloalkyl can be divided, preferably dicyclo or tricyclo, more preferably 3 yuan / 4 yuan, 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 4 yuan, 5 yuan / 5 yuan, 5 yuan / 6 yuan, 6 yuan / 3 yuan, 6 yuan / 4 yuan, 6 yuan / 5 yuan and 6 yuan / 6 yuan of dicycloalkyl. The limiting examples of condensed cycloalkyl include:

[0095]

[0096] The term "bridged cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, and which may contain one or more double bonds. Preferably, it is 6 to 14 members, and more preferably, it is 7 to 10 members (e.g., 7, 8, 9, or 10 members). According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl, preferably a bicyclic, tricyclic, or tetracyclic, more preferably a bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl include:

[0097]

[0098] The cycloalkyl ring includes a cycloalkyl group as described above (including a monocyclic ring, a spirocyclic ring, a condensed ring and a bridged ring) fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring connected to the parent structure is a cycloalkyl group, non-limiting examples include etc.; preferred

[0099] The cycloalkyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0100] The term "alkoxy" refers to -O-(alkyl) and -O-(cycloalkyl), wherein alkyl and cycloalkyl are as defined above. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy and butoxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, it is preferably one or more of the following groups, which are independently selected from D atoms, halogen, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0101] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic substituent containing 3 to 20 ring atoms, one or more of which is a heteroatom selected from nitrogen, oxygen and sulfur, the sulfur being optionally oxo-substituted (i.e. forming a sulfoxide or sulfone), but excluding the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms being carbon. Preferably, it contains 3 to 12 (e.g. 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) ring atoms, of which 1 to 4 (e.g. 1, 2, 3 and 4) are heteroatoms; more preferably, it contains 3 to 8 ring atoms (e.g. 3, 4, 5, 6, 7 and 8), of which 1-3 (e.g. 1, 2 and 3) are heteroatoms; more preferably, it contains 3 to 6 ring atoms, of which 1-3 are heteroatoms; most preferably, it contains 5 or 6 ring atoms, of which 1-3 are heteroatoms. Non-limiting examples of monocyclic heterocyclyls include pyrrolidinyl, tetrahydropyranyl, 1,2,3,6-tetrahydropyridinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclyls include spiro, fused and bridged heterocyclyls.

[0102] The term "spiro heterocyclic group" refers to a polycyclic heterocyclic group of 5 to 20 members, one atom (called spiral atom) shared between monocyclic rings, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, and the sulfur may be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon. It may contain one or more double bonds. It is preferably 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members). According to the number of spiral atoms shared between rings, the spiral heterocyclic group is divided into a monospiro heterocyclic group, a bispiro heterocyclic group or a polyspiro heterocyclic group, preferably a monospiro heterocyclic group and a bispiro heterocyclic group. It is more preferably a 3-yuan / 5-yuan, 3-yuan / 6-yuan, 4-yuan / 4-yuan, 4-yuan / 5-yuan, 4-yuan / 6-yuan, 5-yuan / 5-yuan or 5-yuan / 6-yuan monospiro heterocyclic group. Non-limiting examples of spiral heterocyclic groups include:

[0103]

[0104] The term "fused heterocyclic radical" refers to 5 to 20 yuan, each ring in the system shares a pair of adjacent atoms with other rings in the system. The polycyclic heterocyclic group, one or more rings may contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, and the sulfur may be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan (e.g., 7, 8, 9 or 10 yuan). According to the number of the composition ring, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic radical, preferably a bicyclic or tricyclic, more preferably 3 yuan / 4 yuan, 3 yuan / 5 yuan, 3 yuan / 6 yuan, 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 4 yuan, 5 yuan / 5 yuan, 5 yuan / 6 yuan, 6 yuan / 3 yuan, 6 yuan / 4 yuan, 6 yuan / 5 yuan and 6 yuan / 6 yuan bicyclic fused heterocyclic radical. Non-limiting examples of fused heterocyclic radicals include:

[0105]

[0106] The term "bridged heterocyclic group" refers to a polycyclic heterocyclic group of 5 to 14 members, any two rings sharing two atoms that are not directly connected, which may contain one or more double bonds, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen and sulfur, the sulfur may be optionally oxoed (i.e., forming sulfoxide or sulfone), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, more preferably 7 to 10 members (e.g., 7, 8, 9 or 10 members). According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

[0107]

[0108] The heterocyclic ring includes a heterocyclic group as described above (including a monocyclic, spiro heterocyclic, fused heterocyclic and bridged heterocyclic) fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, non-limiting examples of which include:

[0109] wait.

[0110] The heterocyclyl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0111] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (fused polycyclic is a ring that shares adjacent carbon atom pairs) group with a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. The aryl ring includes an aryl ring as described above fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, and its non-limiting examples include:

[0112]

[0113] The aryl group may be substituted or unsubstituted. When substituted, it may be substituted at any available point of attachment, and the substituents are preferably independently selected from one or more substituents of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0114] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 (e.g., 1, 2, 3, and 4) heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. Heteroaryl is preferably 5 to 10 yuan (e.g., 5, 6, 7, 8, 9, or 10 yuan), more preferably 5 yuan or 6 yuan, such as furanyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidyl, pyrazinyl, pyridazinyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, etc. The heteroaryl ring includes a heteroaryl fused to an aryl, heterocyclyl, or cycloalkyl ring as described above, wherein the ring connected to the parent structure is a heteroaryl ring, and non-limiting examples thereof include:

[0115]

[0116] The heteroaryl group may be substituted or unsubstituted, and when substituted, it may be substituted at any available point of attachment, the substituents preferably being independently selected from one or more of halogen, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyloxy, heterocyclyloxy, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl and heteroaryl.

[0117] The term "amino acid" refers to a molecule containing an amino group and a carboxylic acid group and optionally containing one or more additional groups often referred to as side chains. The amino acids in the sequence of compounds of the present invention are derived from natural amino acids or related amino acid variants and / or derivatives. The abbreviations and codes of the natural amino acids adopt the general rules well known to those in the industry. Amino acids typically use standard single-letter codes (e.g., L = leucine) and α-methyl substituted residues of natural amino acids (e.g., α-methylleucine, or αMeL). Unless explicitly stated, all amino acid residues of the present invention are preferably in the L-configuration, and the D-configuration is also applicable, for example, dS represents D-type Ser.

[0118] Amino acids and their abbreviations and English names are shown in the following table:

[0119]

[0120]

[0121] When used in this article with respect to polypeptides, "amino acid" and "amino acid residue" have the same meaning, which means that when amino acids are connected by chemical bonds, part of their groups are lost due to participation in the formation of the connecting bonds, and the remaining amino acid part is the amino acid residue.

[0122] The term "polypeptide" refers to a sequence of two or more amino acids. A "peptide" may also include an extension of amino acids at the N-terminal and / or C-terminal positions and / or a truncation at the N-terminal and / or C-terminal positions. Typically, amino acid residues may be represented by their full names, their single-letter codes, and / or their three-letter codes. These three methods are completely equivalent.

[0123] The term "protecting group" refers to a group that selectively blocks one reactive site of a polyfunctional compound so that a chemical reaction is selectively carried out at another unprotected reactive site (meaning it is usually associated with it in synthetic chemistry).

[0124] The term "carboxyl protecting group" refers to a carboxyl substituent used to block or protect the functionality of the carboxyl group, and can be converted into a carboxyl protecting group by hydrolysis or deprotection reaction of the carboxyl protecting group. The carboxyl protecting group is preferably an alkyl group (e.g., methyl, ethyl, tert-butyl) or an aralkyl group (e.g., benzyl), and more preferably an amino group, a tert-butoxy group, a methoxy group, or an ethoxy group. In the present invention, a "protected carboxyl" refers to a group formed after a carboxyl group is protected by a suitable carboxyl protecting group, and is preferably a methoxycarbonyl group, an ethoxycarbonyl group, a tert-butoxycarbonyl group, or a benzyloxycarbonyl group. The carboxyl protecting group can be removed by hydrolysis under the catalysis of an acid or a base, and can occasionally be eliminated by a thermal decomposition reaction, for example, the tert-butyl group can be removed under mild acidic conditions, and the benzyl group can be removed by hydrogenolysis. The reagent for removing the carboxyl protecting group is selected from TFA, H2O, LiOH, NaOH, KOH, MeOH, EtOH, and a combination thereof, preferably a combination of TFA and H2O, a combination of LiOH and MeOH, or a combination of LiOH and EtOH. The protected carboxyl group is deprotected to produce the corresponding free acid, and the deprotection is carried out in the presence of a base. The base and the free acid formed by the deprotection form a pharmaceutically acceptable salt.

[0125] In the present invention, "amino protecting group" is equivalent to "amine protecting group" and includes all groups that can be used as conventional amino / amine protecting groups, such as aryl C 1-6 Alkyl, C 1-6 Alkoxy C 1-6 Alkyl, C 1-6 Alkoxycarbonyl, aryloxycarbonyl, C 1-6 Alkylsulfonyl, arylsulfonyl or silyl, etc. The amino protecting group is preferably Ac (acetyl), Boc (tert-butyloxycarbonyl), Moz (p-methoxybenzyloxycarbonyl) or Fmoc (9-fluorenylmethyleneoxycarbonyl). The reagent for removing the amino protecting group is selected from TFA, H2O, LiOH, MeOH, EtOH and a combination thereof, preferably a combination of TFA and H2O, a combination of LiOH and MeOH, or a combination of LiOH and EtOH. The reagent for removing Boc protection is TFA or HCl / EA; preferably TFA. The deprotecting agent used for removing Fmoc protection is a solution of N,N-dimethylformamide (DMF) containing 20% ​​piperidine.

[0126] The term "hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups, wherein alkyl is as defined above.

[0127] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0128] The term "hydroxy" refers to -OH.

[0129] The term "amino" refers to -NH2.

[0130] The term "nitro" refers to -NO2.

[0131] The term "oxo" or "oxo" refers to "=0".

[0132] The term "carboxy" refers to -C(O)OH.

[0133] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0134] "Substituted" means that one or more hydrogen atoms, preferably 1 to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently by a corresponding number of substituents. Those skilled in the art can determine possible or impossible substitutions (by experiment or theory) without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0135] In the present application, the "reaction yield" refers to the yield of the product calculated by detecting the product concentration in the reaction solution by LC-MS after the reaction is completed and before the product is purified.

[0136] The method of the present invention is described below by means of specific examples. It should be understood that these examples are used to illustrate the basic principles, main features and advantages of the present invention, and the present invention is not limited to the scope of the following examples; the implementation conditions adopted in the examples can be further adjusted according to specific requirements, and the implementation conditions not specified are usually the conditions in routine experiments.

[0137] In the following embodiments 1 H NMR spectra were measured using a Bruker instrument (400 MHz), and chemical shifts were expressed in ppm using tetramethylsilane as an internal standard (0.00 ppm). 1 HNMR notation: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, br = broadened, dd = doublet of a doublet, dt = doublet of a triplet. Coupling constants, when given, are given in Hz.

[0138] The mass spectrum was obtained by LC / MS using ESI as the ionization method.

[0139] HPLC model: Agilent 1260, Thermo Fisher U3000; chromatographic column model: Waters xbrigeC18 (4.6*150mm, 3.5μm); mobile phase: A:ACN, B:Water (0.1%H3PO4); flow rate: 1.0mL / min; gradient: 5%Afor 1min, increase to 20%A within 4min, increase to 80%A within 8min, 80%Afor 2min, back to 5%A ​​within 0.1min; wavelength: 220nm; column oven: 35℃.

[0140] TLC: Thin layer chromatography. The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.2mm-0.3mm, and the specification of the thin layer chromatography separation and purification product is 0.4mm-0.5mm.

[0141] Column chromatography generally uses Yantai Huanghai Silica Gel 200-300 mesh silica gel as the carrier.

[0142] In the following examples, unless otherwise specified, all temperatures are in degrees Celsius. Unless otherwise specified, various starting materials and reagents are commercially available or synthesized according to known methods. Commercially available materials and reagents are used directly without further purification. Unless otherwise specified, commercial manufacturers include but are not limited to Sinopharm Group, J&K Technology Co., Ltd., TCI (Shanghai) Chemical Industry Development Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd. and Shanghai Myrel Chemical Technology Co., Ltd.

[0143] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0144] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C-30°C.

[0145] The reaction progress in the embodiment is monitored by thin layer chromatography (TLC), and the developing solvent used in the reaction, the system of eluent for column chromatography or the developing solvent system for thin layer chromatography used for purifying the compound includes: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: n-hexane: ethyl acetate; wherein the volume ratio of the solvent varies according to the polarity of the compound, and a small amount of acidic or alkaline reagent, such as acetic acid or triethylamine, can also be added for adjustment.

[0146] Example 1

[0147]

[0148] In an open glass bottle, a stirring bar, boronic acid derivative 1 (0.05mmol, 1 equivalent), imidazole (0.10mmol, 2 equivalents), copper dichlorophenanthroline (0.001mmol, 0.2 equivalents) and 2mL of methanol were added. After stirring for 3 minutes, 2 equivalents of DIPEA were added and reacted at 25°C for 10 hours to obtain compound 3. The reaction was monitored by LC-MS. After the reaction, the solvent was removed by a rotary evaporator and purified by a silica gel column using a dichloromethane / methanol system (yield was 70%).

[0149] The preparation methods of Examples 2 to 20 are basically the same as those of Example 1, except that the type of catalyst is different from that of Example 1. For specific information, see Table 1. In addition, the reaction time of Example 8 is also different from that of Example 1, and the reaction time of Example 8 is 16 hours.

[0150] The preparation methods of Examples 21 to 27 are basically the same as those of Example 1, except that the types of solvents are different from those of Example 1. For specific information, see Table 1.

[0151] The preparation methods of Examples 28 to 39 are basically the same as those of Example 1, except that the types of bases are different from those of Example 1. For specific information, see Table 1.

[0152] The preparation method of Example 40 is basically the same as that of Example 1, except that the reaction temperature and time are different from those of Example 1. The reaction temperature of Example 40 is 30°C and the reaction time is 16 hours. For specific information, see Table 1.

[0153] Table 1

[0154] Example Catalyst Solvent Base Temperature (°C) Yield (%) 1 [Cu(L6)] MeOH DIPEA 25 70 2 CuO MeOH DIPEA 25 3 3 <![CDATA[Cu2O]]> MeOH DIPEA 25 0 4 <![CDATA[Cu(SO3CF3)2]]> MeOH DIPEA 25 14 5 <![CDATA[Cu(COCF3)2]]> MeOH DIPEA 25 8 6 <![CDATA[Cu[N(SO2CF3)2]2]]> MeOH DIPEA 25 18 7 <![CDATA[Cu(HCOO)2·4H2O]]> MeOH DIPEA 25 34 8 <![CDATA[CuSO4]]> MeOH DIPEA 25 33 9 <![CDATA[Cu(OAc)2]]> MeOH DIPEA 25 22 10 <![CDATA[CuCl2]]> MeOH DIPEA 25 14 11 CuCl MeOH DIPEA 25 14 12 <![CDATA[CuBr2]]> MeOH DIPEA 25 6 13 CuBr MeOH DIPEA 25 10 14 [Cu(L1)] MeOH DIPEA 25 17 15 [Cu(L2)] MeOH DIPEA 25 14 16 [Cu(L3)] MeOH DIPEA 25 33 17 [Cu(L4)] MeOH DIPEA 25 12 18 [Cu(L5)] MeOH DIPEA 25 10 19 [Cu(L7)] MeOH DIPEA 25 trace 20 [Cu(L8)] MeOH DIPEA 25 trace 21 [Cu(L6)] EtOH DIPEA 25 19 22 [Cu(L6)] i-PrOH DIPEA 25 5 23 [Cu(L6)] DMA DIPEA 25 trace 24 [Cu(L6)] THF DIPEA 25 5 25 [Cu(L6)] DMF DIPEA 25 10 26 [Cu(L6)] DCM DIPEA 25 5 27 [Cu(L6)] MeCN DIPEA 25 48 28 [Cu(L6)] MeOH pyridine 25 36 29 [Cu(L6)] MeOH <![CDATA[Na2CO3]]> 25 34 30 [Cu(L6)] MeOH DABCO 25 30 31 [Cu(L6)] MeOH TEMED 25 57 32 [Cu(L6)] MeOH MeONa 25 52 33 [Cu(L6)] MeOH t-BuOK 25 25 34 [Cu(L6)] MeOH 2,6-lutidine 25 30 35 [Cu(L6)] MeOH <![CDATA[K3PO4]]> 25 26 36 [Cu(L6)] MeOH <![CDATA[B(OH)3]]> 25 17 37 [Cu(L6)] MeOH TEA 25 64 38 [Cu(L6)] MeOH <![CDATA[NaHCO3]]> 25 57 39 [Cu(L6)] MeOH / 25 trace 40 [Cu(L6)] MeOH DIPEA 30 86

[0155] Embodiment 41

[0156]

[0157] Boric acid derivative 1 and imidazole or its derivative were reacted under conditions basically the same as those in Example 40, except that the reaction time was 24 hours, to prepare compounds 5 to 10, 12, 14, 15, 17, 19, 20 and compound 23.

[0158] Boric acid derivative 2 and imidazole were selected and reacted under conditions basically the same as those in Example 40, except that the reaction time was 24 hours, to prepare compounds 4 and 22.

[0159] Boric acid derivative 1 and different types of imidazole derivatives were selected and reacted under conditions basically the same as in Example 40, except that the reaction time was 24 hours and the temperature was 40°C to prepare compounds 13, 16, 18, and 21.

[0160] Boric acid derivative 1 and imidazole derivative were selected and reacted under conditions basically the same as those in Example 40, except that the reaction time was 72 hours and the temperature was 40°C to prepare compound 11.

[0161] The structures of different types of imidazole derivatives are as follows:

[0162]

[0163] The structure of boronic acid derivative 2 is The yields of compounds 4 to 23 are shown in Table 2.

[0164] Table 2

[0165]

[0166]

[0167]

[0168] Embodiment 42

[0169]

[0170] Different types of boronic acid derivatives were selected and reacted with imidazole under conditions substantially consistent with those in Example 40 to produce compounds 24 to 40. The specific operation is as follows:

[0171] In an open glass bottle, add a stirrer, a boronic acid derivative (0.05mmol, 1 equivalent), imidazole (0.10mmol, 2 equivalents), copper dichlorophenanthroline (0.001mmol, 0.2 equivalents) and 2mL of methanol. After stirring for 3 minutes, add 2 equivalents of DIPEA and react at 40°C for 24 hours. LC-MS is used to monitor the reaction. After the reaction, the solvent is removed by a rotary evaporator, and the product is purified by preparative HPLC (water containing 1 / 1000 trifluoroacetic acid and acetonitrile as mobile phase).

[0172] The structures of different types of boronic acid derivatives are as follows:

[0173]

[0174]

[0175] The yields of compounds 24-40 are shown in Table 3.

[0176] Table 3

[0177]

[0178]

[0179]

[0180] Embodiment 43

[0181]

[0182] Medium and short oligopeptides of different lengths were selected and reacted with imidazole under conditions substantially consistent with those of Example 40 to generate compounds 41 to 51. The specific operation is as follows:

[0183] In an open glass bottle, add a stirrer, a boronic acid derivative (0.05mmol, 1 equivalent), imidazole (0.10mmol, 2 equivalents), copper dichlorophenanthroline (0.001mmol, 0.2 equivalents) and 2mL of methanol. After stirring for 3 minutes, add 4 equivalents of DIPEA and react at 40°C for 24 hours. LC-MS is used to monitor the reaction. After the reaction, the solvent is removed by a rotary evaporator, and the product is purified by preparative HPLC (water containing 1 / 1000 trifluoroacetic acid and acetonitrile as mobile phase).

[0184] The structures of different types of boronic acid derivatives are as follows:

[0185]

[0186]

[0187] The yields of compounds 41-51 are shown in Table 4.

[0188] Table 4

[0189]

[0190]

[0191]

[0192] Embodiment 44

[0193]

[0194] Some complex peptides were selected and reacted with imidazole under conditions basically consistent with those in Example 43 to generate compounds 52-53. The specific operation is as follows:

[0195] In an open glass bottle, add a stirrer, a boronic acid derivative (0.05mmol, 1 equivalent), imidazole (0.10mmol, 2 equivalents), copper dichlorophenanthroline (0.001mmol, 0.2 equivalents) and 2mL of methanol. After stirring for 3 minutes, add 4 equivalents of DIPEA and react at 40°C for 24 hours. LC-MS is used to monitor the reaction. After the reaction, the solvent is removed by a rotary evaporator, and the product is purified by preparative HPLC (water containing 1 / 1000 trifluoroacetic acid and acetonitrile as mobile phase). The ESIMass spectra of purified compounds 52 and 53 are shown in Figure 1 and Figure 2 .

[0196] The structures 52-53 of different types of boronic acid derivatives and their yields are shown in Table 5 below:

[0197]

[0198]

[0199] The yields of compounds 52-53 are shown in Table 5.

[0200] Table 5

[0201]

[0202] It should be understood that the above embodiments are exemplary and are not intended to include all possible implementations included in the claims. Various modifications and changes may be made on the basis of the above embodiments without departing from the scope of the present disclosure. Similarly, the various technical features of the above embodiments may be arbitrarily combined to form other embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only express several implementations of the present invention and do not limit the scope of protection of the patent of the present invention.

Claims

1. A method for preparing a compound represented by formula (III), characterized in that: The compound represented by formula (I) and the compound represented by formula (II) react in the presence of a copper catalyst to obtain a compound represented by formula (III), wherein R1 is selected from hydrogen, halogen, nitro, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R2, R3, and R4 are each independently selected from hydrogen, halogen, nitro, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl; or, R2 and R3 are linked together with the carbon atom to which they are attached to form a substituted or unsubstituted aryl, heteroaryl or heterocyclic group; The substitution is by at least one selected from hydrogen, deuterium, halogen, cyano, hydroxyl, amino, ester, substituted or unsubstituted alkyl, substituted or unsubstituted haloalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted alkoxy, substituted or unsubstituted haloalkoxy, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl and substituted by a substituent; R5 is hydrogen or an amino protecting group.

2. The method according to claim 1, characterized in that The copper catalyst is selected from one or more of Cu(OAc)2, CuCl, CuCl2, CuBr, CuBr2, Cu(SO3CF3)2, Cu(HCOO)2·4H2O, CuSO4, [Cu(L1)], [Cu(L2)], [Cu(L3)], [Cu(L4)], [Cu(L5)], [Cu(L6)], [Cu(L7)] and [Cu(L8)]; wherein the structural formulas of [Cu(L1)], [Cu(L2)], [Cu(L3)], [Cu(L4)], [Cu(L5)], [Cu(L6)], [Cu(L7)] and [Cu(L8)] are as follows: Preferably, the copper catalyst is selected from one or more of [Cu(L1)], [Cu(L2)], [Cu(L3)], [Cu(L4)], [Cu(L5)], [Cu(L6)], [Cu(L7)], and [Cu(L8)]; More preferably, the copper catalyst is selected from [Cu(L6)].

3. The method according to claim 1, characterized in that The method is carried out in the presence of a solvent, wherein the solvent is selected from one or more of MeOH, DMF, DCM, MeCN, i-PrOH, EtOH, DMA and THF; Preferably, the solvent is selected from one or more of MeOH, MeCN, and EtOH; More preferably, the solvent is selected from MeOH.

4. The method according to claim 1, characterized in that: The method is carried out in the presence of a base, wherein the base is selected from an organic base or an inorganic base; Preferably, the organic base is selected from one or more of TEA, DIPEA, DABCO, TEMED, MeONa, t-BuOK, 2,6-Lutidine, and Py; more preferably, one or more of TEA, DIPEA, TEMED, and MeONa; Preferably, the inorganic base is selected from one or more of NaHCO3, Na2CO3, K3PO4; more preferably, NaHCO3.

5. The method according to claim 1, characterized in that The compound represented by formula (I) is a compound represented by formula (I-1): Wherein, R6 is selected from a peptide chain or an amino protecting group; R7 is selected from a peptide chain, a carboxyl protecting group, a hydroxyl group and an amino group.

6. The method according to claim 5, characterized in that R6 is a peptide chain, and the peptide chain is formed by connecting amino acid residues; Preferably, the amino acid is selected from one or more of substituted or unsubstituted Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro, Lys, Trp, Asp, Gly, Ser, Glu, Thr, His; preferably one or more of Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro; more preferably Tyr; The substitution is substituted by at least one substituent selected from amino or Boc; Alternatively, R6 is an amino protecting group, and the amino protecting group is selected from Ac and / or Boc.

7. The method according to claim 5, characterized in that R7 is selected from a peptide chain, wherein the peptide chain is formed by connecting amino acid residues; Preferably, the amino acid is selected from one or more of substituted or unsubstituted Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro, Lys, Trp, Asp, Asn, Gly, Ser, Glu, Gln, Thr, His; more preferably, one or more of substituted or unsubstituted Tyr, Ala, Val, Leu, Ile, Met, Phe, Pro, Lys, Trp, Asp, Gly, Ser, Glu, Thr, His; The substitution is substituted by at least one substituent selected from amino or Boc; Preferably, the R7 is a carboxyl protecting group, and the carboxyl protecting group is selected from methoxy.

8. The method according to claim 5, characterized in that The compound represented by formula (I-1) is any one of the following:

9. The method according to claim 1, characterized in that: R2, R3, and R4 are each independently selected from hydrogen, halogen, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 3～8 Cycloalkyl, substituted or unsubstituted 3-7 membered heterocyclic group, substituted or unsubstituted C 6～10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; preferably selected from hydrogen, halogen, nitro, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 6～10 Aryl; more preferably selected from hydrogen, halogen, nitro, substituted or unsubstituted C 1-3 Alkyl, substituted or unsubstituted C 6～10 Aryl; Alternatively, R2 and R3 and the carbon atom to which they are attached are linked together to form a substituted or unsubstituted C 6-10 Aryl, 5-10 membered heteroaryl or 3-7 membered heterocyclic group; preferably substituted or unsubstituted C 6-10 Aryl; The substitution is by at least one selected from hydrogen, deuterium, halogen, hydroxyl, amino, C 1～6 Alkyl, C 1～6 Haloalkyl, C 3～8 Cycloalkyl, 3-7 membered heterocyclic group, C 1～6 Alkoxy, C 1～6 Halogenated alkoxy, C 6～10 Aryl, 5- to 6-membered heteroaryl and Substituents; preferably selected from hydrogen, deuterium, halogen, amino and substituted by a substituent; R5 is an amino protecting group.

10. The method according to claim 9, characterized in that The compound represented by formula (II) is any one of the following:

Images