Cyclic peptide compound and use thereof

Abstract

The present invention relates to a cyclic peptide compound and the use thereof. The cyclic peptide compound has a structure as represented by formula (I), or is a stereoisomer, tautomer, nitrogen oxide, solvate, metabolite, pharmaceutically acceptable salt or prodrug of the structure as represented by formula (I). Provided is a pharmaceutical composition containing the cyclic peptide compound. The cyclic peptide compound or pharmaceutical composition is used in the preparation of a drug for preventing, treating, alleviating, or modulating diseases associated with or mediated by a melanocortin receptor.

Description

A cyclic peptide compound and its uses Technical Field

[0001] This invention relates to a cyclic peptide compound and its uses, particularly its use in the preparation of medicaments for the prevention, treatment, relief or regulation of diseases mediated by melanocortin receptors. Background Technology

[0002] Melanocortin receptors (MCRs) influence inflammation, immune system responses, metabolism, food intake, and sexual function. There are five types of melanocortin receptors in the human body, named MC1R to MC5R. Significant pharmacological effects can be achieved by modulating these receptors using receptor-specific agonists that activate their function or receptor-specific antagonists that block their function. Many tissues and immune cells in the eye (as well as in other sites such as the intestines and kidneys) express melanocortin receptors, allowing drugs the opportunity to directly activate these naturally occurring pathways to address disease inflammation.

[0003] Melanocortin receptors and their subtypes include: 1) melanocortin receptor 1 (MC1R), expressed on normal human melanocytes, melanoma cells, macrophages, and other cells, which is involved in inflammation-mediated processes, hair and skin pigmentation, and other functions; 2) melanocortin receptor 2 (MC2R), expressed on adrenal cells, which mediates steroid production; 3) melanocortin receptors 3 and 4 (MC3R and MC4R), expressed on cells in the hypothalamus, midbrain, brainstem, and peripheral tissues, which are involved in energy homeostasis, feeding behavior, inflammation-mediated processes, and other functions; and 4) melanocortin receptor 5 (MC5R), expressed in widely distributed peripheral tissues, which is involved in the regulation of the exocrine gland system and other functions.

[0004] Despite extensive research and numerous patent applications filed by pharmaceutical companies and medical scientists regarding melanocortin receptor-specific peptides, only bremelanotide (trade name) has been approved in the United States. For sale, for use in premenopausal women with decreased libido; Amerin (under brand name) Three drugs are available for sale, including one for preventing phototoxicity in adult patients with erythropoietic protoporphyria. Currently, there are no drugs on the market for melanocortin receptor-mediated ophthalmic diseases, indicating an unmet clinical need. Summary of the Invention

[0005] In a first aspect, the present invention provides a cyclic peptide compound having a structure as shown in Formula (I), or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of Formula (I).

[0006] Among them, R 1 It can be hydrogen, deuterium, alkyl, haloalkyl, heteroalkyl, or cycloalkyl;

[0007] R 2 It can be hydrogen, deuterium, alkyl, heterocyclic, or heteroaryl;

[0008] R 3 and R 4 Each is independently an aryl or heteroaryl group;

[0009] R 5 It can be amino, alkyl, alkoxy, heteroaryl, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3, -S(=O)-CH3 or -C(=O)-OR 9 ;

[0010] R 6 Aryl, heteroaryl, heterocyclic, or cycloalkyl;

[0011] R 7 and R 8 Each of these can be independently hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, alkenylalkyl, cycloalkylalkyl, haloalkoxy, cycloalkyloxy, heterocyclicoxy, cycloalkyl, heterocyclic, aryl, or heteroaryl.

[0012] Optionally, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles;

[0013] R 9 It can be hydrogen, alkyl, haloalkyl, deuterated alkyl, hydroxyalkyl, or cycloalkyl;

[0014] R 10 It can be hydrogen, deuterium, halogen, hydroxyl, alkyl, haloalkyl, deuterated alkyl, hydroxyalkyl, or cycloalkyl;

[0015] A is -(CH2) p -or-(CH2) q -O-(CH2)u -;

[0016] m, n, t, p, q or u are each independently 0, 1, 2, 3, 4 or 5;

[0017] R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 The groups described in A are independently and optionally replaced by 1, 2, 3 or 4 substituents selected from deuterium, halogen, amino, hydroxyl, cyano, oxo, nitro, alkyl, alkoxy, alkenyl, alkynyl, haloalkyl, cyanoalkyl, alkoxyalkyl, cycloalkylalkyl, heteroalkyl, heteroalkylalkyl, alkenylalkyl, alkynylalkyl, aryl, heteroaryl, heterocyclic, and cycloalkyl.

[0018] In some embodiments, the cyclic peptide compound is described, wherein,

[0019] R 5 For amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-9 heteroaryl, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3, -S(=O)-CH3 or -C(=O)-OR 9 ;

[0020] R 6 C 6-10 Aryl, C 1-9 heteroaryl, C 1-9 Heterocyclic group or C 3-8 cycloalkyl;

[0021] R 9 For hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl or C 3-8 Cycloalkyl.

[0022] In some other embodiments, the cyclic peptide compound, wherein,

[0023] R 5 It is an amino group, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3 or -S(=O)-CH3;

[0024] R 6 C 6-10 Aryl or C 1-9 heteroaryl; preferably

[0025] In some embodiments, the compound has a structure as shown in formula (II), or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of a compound with a structure as shown in formula (II).

[0026] Among them, R 1 R 2 R 3 R 4 R 7 R 8 A, m, and n are defined in equation (I).

[0027] In some embodiments, the cyclic peptide compound, wherein,

[0028] Among them, R 1 For hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Heteroalkyl or C 3-8 cycloalkyl;

[0029] R 2 For hydrogen, deuterium, C 1-6 Alkyl, C 1-9 Heterocyclic groups or independently optionally halogenated, C 1-4 Alkyl-substituted C 1-9 Mixed aromatic compounds.

[0030] In some other embodiments, the cyclic peptide compound, wherein, R 1 C is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, or optionally substituted with 1, 2, 3, 4, 5, or 6 F, Cl, Br, or OH groups. 1-4 alkyl;

[0031] R 2 C 1-4 Alkyl group, independently and optionally with 1, 2, or 3 halogens or with C 1-4 Alkyl-substituted pyrrole, pyrazol, imidazole, or triazolyl groups.

[0032] In some embodiments, the cyclic peptide compound, wherein R 3 and R 4Each independently can be halogenated or C 1-4 Alkyl-substituted C 6-10 Aryl or C 1-9 Mixed aromatic compounds.

[0033] In some other embodiments, the cyclic peptide compound, wherein R 3 and R 4 Each can be independently selected by 1, 2, or 3 F, Cl, Br, or C atoms. 1-4 Alkyl-substituted pyrrole, pyrazolyl, imidazolyl, triazolyl, phenyl, naphthyl, pyridyl, or C 5-9 Dense heteroaryl compounds.

[0034] In some other embodiments, the cyclic peptide compound, wherein R 4 for

[0035] In some embodiments, the cyclic peptide compound, wherein R 7 and R 8 Each of these can be independently represented by hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, or C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Heteroalkyl, C 1- 6-alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy C 1-6 Alkyl, C 2-6 alkenyl C 1-6 Alkyl, C 3-6 cycloalkyl C 1-6 Alkyl, C 1-6 Halogenated alkoxy groups, C 3-6 Cycloalkyloxy, C 1-6 Heterocyclic oxygen, C 3-6 cycloalkyl, C 1-6 Heterocyclic group, C 6-10 Aryl or C 1-6 heteroaryl; optionally, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles;

[0036] R 10 Hydrogen, deuterium, halogen, hydroxyl, C 1-3 Alkyl, C 1-3 Haloalkyl, C1-3 Deuterated alkyl, C 1-3 Hydroxyalkyl or C 3- 6-Cycloalkyl.

[0037] In some other embodiments, the cyclic peptide compound, wherein R 7 and R 8 Each of these can be independently represented by hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, or C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Heteroalkyl, C 1-3 Alkoxy, C 1-3 Hydroxyalkyl or C 1-3 Alkoxy C 1-3 Alkyl; alternatively, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles;

[0038] R 10 For hydrogen, deuterium, halogen, hydroxyl, methyl, ethyl, C substituted with 1, 2, or 3 F or Cl atoms 1-3 Alkyl, C 1- 3-Deuterated alkyl, C 1-3 Hydroxyalkyl or C 3-6 Cycloalkyl.

[0039] In a second aspect, the present invention also provides a pharmaceutical composition comprising the above-mentioned cyclic peptide compound, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, medium, or combination thereof.

[0040] Thirdly, the use of the cyclic peptide compound or the pharmaceutical composition in the preparation of a medicament for the prevention, treatment, relief or regulation of diseases mediated by melanocortin receptors.

[0041] In some embodiments, the disease described in the intended use is an ocular disease, selected from one or more of the following: dry eye, corneal ulcer, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scarring, epithelial defects, keratoconjunctivitis, idiopathic uveitis, corneal transplantation, age-related macular degeneration, diabetic retinopathy, blepharitis, glaucoma, ocular hypertension, postoperative eye pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, and choroidal neovascularization. Vascular occlusive diseases, allergic eye diseases, tumors, retinitis pigmentosa, ocular infections, scleritis, ptosis, miosis, eye pain, mydriasis, neuralgia, cicatricial ocular surface diseases, ocular infections, inflammatory eye diseases, ocular surface diseases, corneal diseases, retinal diseases, ocular manifestations of systemic diseases, hereditary ocular diseases, ocular tumors, increased intraocular pressure, herpes infections, pterygium, wounds extending to the ocular surface, post-refractive keratotomy ocular pain and inflammation, corneal thermal or chemical burns, scleral wounds, keratoconus or conjunctival wounds.

[0042] Fourthly, a method for treating melanocortin receptor-mediated diseases, indications, symptoms, or syndromes in humans or non-human mammals, the method comprising administering the cyclic peptide compound of the present invention or the pharmaceutical composition of the present invention.

[0043] In some embodiments, the method wherein the melanocortin-mediated disease is an ocular disease selected from one or more of the following: dry eye, corneal ulcer, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scarring, epithelial defects, keratoconjunctivitis, idiopathic uveitis, corneal transplantation, age-related macular degeneration, diabetic retinopathy, blepharitis, glaucoma, ocular hypertension, postoperative ocular pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascularization. Vascular membranes, vascular occlusive diseases, allergic eye diseases, tumors, retinitis pigmentosa, ocular infections, scleritis, ptosis, miosis, eye pain, mydriasis, neuralgia, cicatricial ocular surface diseases, ocular infections, inflammatory eye diseases, ocular surface diseases, corneal diseases, retinal diseases, ocular manifestations of systemic diseases, hereditary ocular diseases, ocular tumors, increased intraocular pressure, herpes infections, pterygium, wounds extending to the ocular surface, post-refractive keratotomy ocular pain and inflammation, corneal thermal or chemical burns, scleral wounds, keratoconus or conjunctival wounds.

[0044] The foregoing description only outlines certain aspects of the invention, but is not limited to these aspects. These and other aspects will be described in more detail below. Detailed Implementation

[0045] Definitions and general terms

[0046] Unless otherwise stated, the terms used in the specification and claims of this invention have the following definitions.

[0047] Certain embodiments of the invention will now be described in detail, examples of which are illustrated by the accompanying structural and chemical formulas. The invention is intended to cover all alternatives, modifications, and equivalents, all of which are included within the scope of the invention as defined in the claims. Those skilled in the art will recognize that many similar or equivalent methods and materials can be used to practice the invention. The invention is by no means limited to the methods and materials described herein. In the event that one or more of the incorporated documents, patents, and similar materials differ from or contradict this application (including, but not limited to, defined terminology, application of terminology, described techniques, etc.), this application shall prevail.

[0048] It should be further appreciated that certain features of the invention, for clarity, have been described in multiple independent embodiments, but may also be provided in combination in a single embodiment. Conversely, various features of the invention, for brevity, have been described in a single embodiment, but may also be provided individually or in any suitable sub-combination.

[0049] Unless otherwise stated, all technical terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. All patents and publications related to this invention are incorporated herein by reference in their entirety.

[0050] Unless otherwise stated, the following definitions shall apply as used herein. For the purposes of this invention, chemical elements are consistent with the periodic table (CAS edition) and the Handbook of Chemistry and Physics, 75th edition, 1994. Furthermore, general principles of organic chemistry can be found in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March's Advanced Organic Chemistry” by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, the entire contents of which are incorporated herein by reference.

[0051] Unless otherwise stated or there is a clear conflict in the context, the articles “a,” “an,” and “described” as used herein are intended to include “at least one” or “one or more.” Therefore, these articles as used herein refer to articles for one or more (i.e., at least one) objects. For example, “a component” refers to one or more components, meaning that more than one component may be considered for use or adoption in the implementation of the described embodiments.

[0052] The term "patient" as used in this invention refers to a person (including adults and children) or other animal. In some embodiments, "patient" refers to a person.

[0053] The term "subject" as used in this invention refers to a mammal or other animal. In some embodiments, "subject" refers to a mammal. In some embodiments, "subject" refers to a human (including adults and children).

[0054] The term "comprising" is an open-ended expression, meaning it includes the contents specified in this invention, but does not exclude other aspects.

[0055] "Stereoisomers" are compounds that have the same chemical structure but whose atoms or groups are arranged differently in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and hindered isomers, etc.

[0056] A diastereomer is a stereoisomer that has two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and reactivity. Mixtures of diastereomers can be separated by high-resolution analytical procedures such as electrophoresis and chromatography, for example, HPLC.

[0057] The stereochemical definitions and rules used in this invention generally follow those of S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., New York, 1994.

[0058] Many organic compounds exist in an optically active form, meaning they possess the ability to rotate the plane of plane-polarized light. In describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to one or more of its chiral centers. The prefixes d and l or (+) and (-) are symbols used to specify the plane-polarized light rotation caused by the compound, where (-) or l indicates that the compound is levorotatory. Compounds with the prefix (+) or d are dextrorotatory. A specific stereoisomer is an enantiomer, and a mixture of such isomers is called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which can occur when there is no stereoselectivity or stereospecificity in the chemical reaction or process.

[0059] Any asymmetric atom (e.g., carbon, etc.) in the compounds disclosed in this invention can exist in a racemic or enantiomerically enriched form, such as in (R)-, (S)-, or (R,S)- configurations. In some embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R)- or (S)- configuration.

[0060] Depending on the choice of starting materials and methods, the compounds of this invention can exist as one or a mixture of possible isomers, such as racemic mixtures and diastereomeric mixtures (depending on the number of asymmetric carbon atoms). Optically active (R)- or (S)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. If the compound contains a double bond, the substituents may be E or Z configurations; if the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituents may be cis or trans configurations.

[0061] Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0062] Unless otherwise indicated, the structural formulas described in this invention include all isomers (e.g., enantiomers, diastereomers, and geometric isomers (or conformational isomers): for example, R and S configurations containing an asymmetric center, (Z) and (E) isomers of double bonds, and (Z) and (E) conformational isomers. Therefore, any single stereochemical isomer of the compounds of this invention, or a mixture of its enantiomers, diastereomers, or geometric isomers (or conformational isomers), is within the scope of this invention.

[0063] As used in this invention, the term "prodrug" refers to the conversion of a compound into the compounds represented by the general formulas of this invention in vivo. Such conversion is influenced by the hydrolysis of the prodrug in the blood or its enzymatic conversion into the parent structure in the blood or tissues. The prodrug compounds of this invention can be esters; in existing inventions, esters that can serve as prodrugs include phenyl esters, aliphatic (C1-24) esters, acyloxymethyl esters, carbonates, carbamates, and amino acid esters. For example, a compound in this invention contains a hydroxyl group, meaning it can be acylated to obtain the prodrug form. Other prodrug forms include phosphate esters, such as those obtained by phosphorylation of a hydroxyl group on the parent compound. For a complete discussion of prodrugs, please refer to the following literature: T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series; Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987; J. Rautio et al, Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270; and SJ Hecker et al, Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51, 2328-2345.

[0064] Racemic mixtures of any resulting end product or intermediate can be separated into optical enantiomers using known methods, such as by separating their diastereomeric salts. Racemic products can also be separated by chiral chromatography, such as high-performance liquid chromatography (HPLC) using chiral adsorbents. In particular, enantiomers can be prepared by asymmetric synthesis, for example, see Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis (2nd Ed. Robert E. Gawley, Jeffrey Aubé, Elsevier, Oxford, UK, 2012); Eliel, ELStereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); Wilen, SH Tables of Resolving Agents and Optical Resolutions p. 268 (ELEliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972); Chiral Separation Techniques: A Practical Approach (Subramanian, G. Ed., Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany, 2007).

[0065] The terms "tautomer" or "tautomer form" refer to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (also called prototropic tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions via the rearrangement of some bonding electrons. A specific example of a keto-enol tautomer is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one. Another example of tautomerism is phenol-keto tautomerism. A specific example of a phenol-keto tautomer is the interconversion between pyridine-4-ol and pyridine-4(1H)-keto. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0066] The salts mentioned in this invention are pharmaceutically acceptable salts, and the term "pharmaceutically acceptable salts" is well known in the field, as described in the literature: Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmacol Sci, 1997, 66, 1-19. Pharmaceutically acceptable, non-limiting examples of salts include inorganic acid salts formed by reactions with amino groups, such as hydrochlorides, hydrobroms, phosphates, metaphosphates, sulfates, sulfites, nitrates, and perchlorates, and organic acid salts, such as carboxylates, sulfonates, sulfinates, and thiocarboxylates, specifically, but not limited to, methanesulfonates, ethanesulfonates, formates, acetates, succinates, benzoates, succinates, bis(hydroxynaphthyl) salts, salicylates, galactobionates, gluconates, mandelates, 1,2-ethanedisulfonates, 2-naphthalenesulfonates, carbonates, trifluoroacetates, glycolates, hydroxyethylsulfonates, oxalates, maleates, tartrates, citrates, succinates, malonates, benzenesulfonates, p-toluenesulfonates, malates, fumarates, lactates, lactobionates, or oxalic acid, or obtained by other methods described in the literature, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecyl sulfate, ethanesulfonate, glucono-heptahydrate, glycerophosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodate, 2-hydroxy-ethanesulfonate, lacturonate, laurate, lauryl sulfate, nicotinate, nitrate, oleate, palmitate, pyruvate, pectate, persulfate, 3-phenylpropionate, picrate, pentanoate, propionate, stearate, thiocyanate, undecanoate, valerate, etc. Furthermore, pharmaceutically acceptable salts also include those obtained by means of appropriate bases, such as alkali metals, alkaline earth metals, ammonium, and N+(C) salts. 1-4 Salts of alkyl groups (4). This invention also contemplates quaternary ammonium salts formed from any compound containing an N group. Water-soluble or oil-soluble or dispersed products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts, and amine cations resistant to the formation of equilibrium ions, such as halides, carboxylates, sulfates, phosphates, nitrates, C 1- 8. Sulfonates and aromatic sulfonates.

[0067] Medicinal salts can form with inorganic and organic acids, such as acetates, aspartates, benzoates, benzenesulfonates, bromides / hydrobromoates, bicarbonates / carbonates, hydrogen sulfates / sulfates, camphor sulfonates, chlorides / hydrochlorides, theophylline salts, citrates, ethanedisulfonates, fumarates, gluconate, gluconate, glucuronide, hippurate, hydroiodide / iodide, hydroxyethyl sulfonate, lactates, lacturonide, lauryl sulfate, malates, maleates, malonates, mandelates, methanesulfonates, methyl sulfates, naphthates, naphthalenesulfonates, nicotinates, nitrates, stearates, oleates, oxalates, palmitates, pyrates, phosphates / hydrogen phosphates / dihydrogen phosphates, polygalactosates, propions, stearates, succinates, sulfosalicylates, tartrates, toluenesulfonates, and trifluoroacetates. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid.

[0068] Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, sulfosalicylic acid, etc.

[0069] In this invention, "solvent" refers to an association formed by one or more solvent molecules and the compound of this invention. Solvents forming solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed when the solvent molecules are water.

[0070] The term "protecting group" or "PG" refers to a substituent that, when reacting with other functional groups, is typically used to block or protect specific functionalities. For example, "amino protecting group" refers to a substituent attached to an amino group to block or protect the functionality of the amino group in a compound. Suitable amino protecting groups include acetyl, trifluoroacetyl, tert-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ), and 9-fluorenemethoxycarbonyl (Fmoc). Similarly, "hydroxyl protecting group" refers to a substituent of a hydroxyl group used to block or protect its functionality; suitable protecting groups include acetyl and silyl. "Carboxyl protecting group" refers to a substituent of a carboxyl group used to block or protect its functionality. Common carboxyl protecting groups include -CH2CH2SO2Ph, cyanoethyl, 2-(trimethylsilyl)ethyl, 2-(trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrobenzenesulfonyl)ethyl, 2-(diphenylphosphine)ethyl, nitroethyl, etc. For a general description of protecting groups, please refer to: TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991; and PJ Kocienski, Protecting Groups, Thieme, Stuttgart, 2005.

[0071] "Pharmaceutical composition" means a salt of one or more of the compounds described herein, or a physiologically / pharmaceutically acceptable salt or prodrug, mixed with other chemical components, such as physiologically / pharmaceutical acceptable carriers or excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compound to a living organism.

[0072] As used in this invention, the term "treatment" refers to any disease or condition, and in some embodiments, it means improving the disease or condition (i.e., slowing down or stopping or alleviating the development of the disease or at least one of its clinical symptoms). In other embodiments, "treatment" means alleviating or improving at least one bodily parameter, including bodily parameters that may not be perceived by the patient. In still other embodiments, "treatment" means regulating the disease or condition physically (e.g., stabilizing perceptible symptoms) or physiologically (e.g., stabilizing bodily parameters) or both. In still other embodiments, "treatment" means preventing or delaying the onset, occurrence, or worsening of the disease or condition.

[0073] Any structural formulas provided in this invention are intended to represent both the unenriched and isotopically enriched forms of these compounds. Isotopically enriched compounds have the structures described by the general formulas provided in this invention, except that one or more atoms are replaced by atoms having a chosen atomic weight or mass number. Exemplary isotopes that may be introduced into the compounds of this invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as... 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 17 O, 18 O, 18 F, 31 P, 32 P, 35 S, 36 Cl and 125 I.

[0074] On the other hand, the compounds described in this invention include isotopically enriched compounds as defined in this invention, for example, compounds containing radioactive isotopes, such as... 3 H, 14 C and 18 Those compounds of F, or those containing non-radioactive isotopes, such as 2 H and 13 C. Compounds enriched by this type of isotope can be used for metabolic studies (using...) 14 C) Reaction kinetic studies (using, for example) 2 H or 3 H) Detection or imaging techniques, such as positron emission tomography (PET) or single-photon emission computed tomography (SPECT) which includes the determination of drug or substrate tissue distribution, may be used in the patient's radiotherapy. 18 F-enriched compounds are particularly desirable for PET or SPECT studies. The isotopically enriched compounds of the general formulas of this invention can be prepared using conventional techniques familiar to those skilled in the art, or by replacing the previously used unlabeled reagents with suitable isotopically labeled reagents, as described in the examples and preparation processes of this invention.

[0075] In addition, heavier isotopes, especially deuterium (i.e., 2Substitution with H or D can provide certain therapeutic advantages resulting from increased metabolic stability. For example, this may lead to an increased half-life in vivo, a reduced dose requirement, or an improved therapeutic index. It should be understood that deuterium in this invention is considered a substituent in the compounds represented by the general formulas of this invention. The concentration of such heavier isotopes, particularly deuterium, can be defined using an isotope enrichment factor. As used in this invention, the term "isotope enrichment factor" refers to the ratio between the isotopic abundance of the specified isotope and its native abundance. If the substituent of the compound of the present invention is designated as deuterium, the compound has an isotopic enrichment factor of at least 3500 (52.5% deuterium doping at each designated deuterium atom), at least 4000 (60% deuterium doping), at least 4500 (67.5% deuterium doping), at least 5000 (75% deuterium doping), at least 5500 (82.5% deuterium doping), at least 6000 (90% deuterium doping), at least 6333.3 (95% deuterium doping), at least 6466.7 (97% deuterium doping), at least 6600 (99% deuterium doping), or at least 6633.3 (99.5% deuterium doping) with respect to each designated deuterium atom. The pharmaceutically usable solvates of the present invention include those in which the crystallization solvent may be isotopically substituted, such as D2O, acetone-d6, DMSO-d6.

[0076] As described in this invention, the compounds of this invention may optionally be substituted with one or more substituents, such as the general formula compounds above, or as the specific examples, subclasses, and classes of compounds included in this invention as described in the embodiments. It should be understood that the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted." Generally, the term "optionally," whether or not it precedes the term "substituted," indicates that one or more hydrogen atoms in the given structure are substituted by a specific substituent. Unless otherwise indicated, an optional substituent group may have one substituent substituted at each substituted position of the group. When more than one position in the given structural formula is substituted by one or more substituents selected from a specific group, the substituents may be substituted at the same or different positions. The substituents mentioned therein can be, but are not limited to, deuterium, hydroxyl, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkylamino, alkylthio, alkyl, alkenyl, alkynyl, heterocyclic, mercapto, nitro, aryloxy, heteroaryloxy, oxo (=O), carboxyl, hydroxy-substituted alkoxy, hydroxy-substituted alkyl-C (=O), alkyl-C (=O), alkyl-S (=O), alkyl-S (=O)2-, hydroxy-substituted alkyl-S (=O), hydroxy-substituted alkyl-S (=O)2, carboxyalkoxy, etc.

[0077] As used in this invention, the term "alkyl" refers to a saturated straight-chain or branched monovalent hydrocarbon group having 1-20 carbon atoms, or 1-10 carbon atoms, or 1-8 carbon atoms, or 1-6 carbon atoms, or 1-4 carbon atoms, or 1-3 carbon atoms, wherein the alkyl group may be independently and optionally substituted by one or more substituents described in this invention. Examples of alkyl groups include, but are not limited to, methyl (Me,-CH3), ethyl (Et,-CH2CH3), n-propyl (n-Pr,-CH2CH2CH3), isopropyl (i-Pr,-CH(CH3)2), n-butyl (n-Bu,-CH2CH2CH2CH3), isobutyl (i-Bu,-CH2CH(CH3)2), sec-butyl (s-Bu,-CH(CH3)CH2CH3), tert-butyl (t-Bu, -C(CH3)3), n-pentyl (-CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1- Butyl (-CH2CH(CH3)CH2CH3), n-hexyl (-CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3) ), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), n-heptyl, n-octyl, etc. The term "alkyl" and its prefix "alkane" are used herein to refer to both straight-chain and branched saturated carbon chains. The term "alkane" is used herein to refer to a saturated divalent hydrocarbon group obtained by eliminating two hydrogen atoms from a straight-chain or branched saturated hydrocarbon; examples of such groups include, but are not limited to, methylene, methine, methinepropyl, etc.

[0078] The term "alkoxy" as used in this invention refers to an alkyl group, as defined herein, that is attached to the main carbon chain by an oxygen atom. Examples of such alkyl groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, etc. Furthermore, the alkoxy group may be substituted or unsubstituted, wherein the substituent may be, but is not limited to, hydroxyl, amino, halogen, cyano, alkoxy, alkyl, alkenyl, alkynyl, mercapto, nitro, etc.

[0079] The term "alkenyl" refers to a straight-chain or branched monovalent hydrocarbon group of 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, wherein at least one position is unsaturated, i.e., one C-C is an sp2 double bond, wherein the alkenyl group may be independently and optionally substituted by one or more substituents described in this invention, including groups with "trans", "cis" or "E", "Z" orientations, wherein specific examples of alkenyl include, but are not limited to, vinyl (-CH=CH2), allyl (-CH2CH=CH2), etc.

[0080] The term "alkynyl" refers to a straight-chain or branched monovalent hydrocarbon group with 2-12 carbon atoms, or 2-8 carbon atoms, or 2-6 carbon atoms, or 2-4 carbon atoms, wherein at least one position is unsaturated, i.e., one C C is an sp triple bond, wherein the alkynyl group may be independently and optionally substituted by one or more substituents described in this invention, wherein specific examples of alkynyl include, but are not limited to, ethynyl (-C≡CH), propynyl (-CH2C≡CH), etc.

[0081] The term "cycloalkyl" or "carbocyclic" refers to a monovalent or polyvalent, non-aromatic, saturated or partially unsaturated ring that does not contain heteroatoms, including monocyclic rings of 3-12 carbon atoms or bicyclic rings of 7-12 carbon atoms. Bicyclic carbocyclic rings with 7-12 atoms can be bicyclic [4,5], [5,5], [5,6], or [6,6] systems, while bicyclic carbocyclic rings with 9 or 10 atoms can be bicyclic [5,6] or [6,6] systems. Suitable cyclic aliphatic groups include, but are not limited to, cycloalkyl, cycloalkenyl, and cycloynyl groups. Examples of cyclic aliphatic groups include, but are by no means limited to, cyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopentyl-1-enyl, 1-cyclopentyl-2-enyl, 1-cyclopentyl-3-enyl, cyclohexyl, 1-cyclohexyl-1-enyl, 1-cyclohexyl-2-enyl, 1-cyclohexyl-3-enyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, cyclododecyl, etc. Furthermore, the "cyclic aliphatic group" or "carbocyclic", "carbocyclic group", and "cycloalkyl" may be substituted or unsubstituted, wherein the substituent may be, but is not limited to, hydroxyl, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkylamino, alkyl, alkenyl, alkynyl, heterocyclic, mercapto, nitro, aryloxy, hydroxy-substituted alkoxy, hydroxy-substituted alkyl-C(=O), alkyl-C(=O), alkyl-S(=O), alkyl-S(=O)2-, hydroxy-substituted alkyl-S(=O), hydroxy-substituted alkyl-S(=O)2, carboxyalkoxy, etc.

[0082] The terms “heterocyclic,” “heterocyclic group,” “heterocyclic alicyclic group,” or “heterocyclic” are used interchangeably herein to refer to monocyclic, bicyclic, or tricyclic systems in which one or more carbon atoms on the ring are independently and optionally substituted with heteroatoms, which have the meaning as described herein. The ring may be fully saturated or contain one or more unsaturations, but is by no means aromatic, and has only one connection point to another molecule. One or more hydrogen atoms on the ring are independently and optionally substituted with one or more substituents described herein. Some of these embodiments are that the "heterocycle", "heterocyclic group", "heterocyclic alicyclic group" or "heterocyclic" group is a 3-7 membered monocyclic ring (1-6 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, wherein S or P is optionally replaced by one or more oxygen atoms to obtain a group such as SO, SO2, PO, PO2, and when the ring is a three membered ring, there is only one heteroatom), or a 7-10 membered bicyclic ring (4-9 carbon atoms and 1-3 heteroatoms selected from N, O, P, S, wherein S or P is optionally replaced by one or more oxygen atoms to obtain a group such as SO, SO2, PO, PO2).

[0083] Heterocyclic groups can be carbonyl or heteroatomyl. "Heterocyclic group" also includes groups formed by the fusion of a heterocyclic group with a saturated or partially unsaturated ring or heterocycle. Examples of heterocycles include, but are not limited to, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiophenyl, piperidinyl, morpholinyl, thiomorpholinyl, thiazolyl, thiazolyl, oxazolyl, piperazine, homopiperazine, aziridine, oxacyclobutyl, thiohexacyclobutyl, piperidinyl, homopiperidinyl, glycidyl, aziridineheptyl, oxacycloheptyl, thiohexacycloheptyl, 4-methoxy-piperidin-1-yl, 1,2,3,6-tetrahydropyridin-1-yl, oxacyclobutyl... 2-diazine Base, sulfur nitrogen 1-pyrrololin-1-yl, 2-pyrrololin-3-pyrrololin-1-yl, dihydroindolyl, 2H-pyranyl, 4H-pyranyl, dioxacyclohexyl, 1,3-dioxopentyl, pyrazolinyl, dithiaalkyl, dithiamonyl, dihydrothiophenyl, pyrazolinyl imidazolinyl, imidazolinyl, 1,2,3,4-tetrahydroisoquinolinyl, 1,2,6-thiadiazinane 1,1-dioxo-2-yl, 4-hydroxy-1,4-azaphosphane 4-oxide-1-yl, 2-hydroxy-1-(piperazin-1-yl)acetone-4-yl, 2-hydroxy-1-(5,6-dihydro-1,2,4-triazin-1(4H)-yl)acetone-4-yl, 5,6-dihydro-4 H-1,2,4-oxadiazine-4-yl, 2-hydroxy-1-(5,6-dihydropyridin-1(2H)-yl) acetone-4-yl, 3-azabicyclo[3.1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, 2-methyl-5,6,7,8-tetrahydro-[1,2,4]triazol[1,5-c]pyrimidin-6-yl, 4,5,6,7-tetrahydroisoxazol[4,3-c]pyridin-5-yl, 3H-indolyl-2-oxo-5-azabicyclo[2.2.1]heptane-5-yl, 2-oxo-5-azabicyclo[2.2.2]octane-5-yl, quinazinyl and N-pyridinyl urea. Examples of heterocyclic groups also include 1,1-dioxothiomorpholino, and those in which two carbon atoms on the ring are replaced by oxygen atoms, such as pyrimidinide groups. Furthermore, the heterocyclic group can be substituted or unsubstituted, wherein the substituent can be, but is not limited to, oxo(=O), hydroxyl, amino, halogen, cyano, heteroaryl, alkoxy, alkylamino, alkyl, alkenyl, alkynyl, heterocyclic, mercapto, nitro, aryloxy, hydroxy-substituted alkoxy, hydroxy-substituted alkyl-C(=O), alkyl-C(=O), alkyl-S(=O), alkyl-S(=O)2-, hydroxy-substituted alkyl-S(=O), hydroxy-substituted alkyl-S(=O)2, carboxyalkoxy, etc.

[0084] The term "aryl" or "aromatic ring" can be used alone or as a part of "aralkyl," "ararylalkoxy," or "aryloxyalkyl," referring to monocyclic, bicyclic, and tricyclic carbocyclic systems containing a total of 6-14 membered rings, wherein at least one ring system is aromatic, and each ring system contains 3-7 membered rings with only one attachment point connected to the rest of the molecule. The term "aryl" can be used interchangeably with the term "aromatic ring," as aromatic rings can include phenyl, naphthyl, and anthracene. Furthermore, the aryl group may be substituted or unsubstituted, wherein the substituent may be, but is not limited to, hydroxyl, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkylamino, alkyl, alkenyl, alkynyl, heterocyclic, mercapto, nitro, aryloxy, hydroxy-substituted alkoxy, hydroxy-substituted alkyl-C(=O), alkyl-C(=O), alkyl-S(=O), alkyl-S(=O)2-, hydroxy-substituted alkyl-S(=O), hydroxy-substituted alkyl-S(=O)2, carboxyalkoxy, etc.

[0085] The term "heteroaryl" or "heteroaromatic ring" refers to monocyclic, bicyclic, and tricyclic systems containing 5-14 membered rings, wherein at least one ring system is aromatic and contains one or more heteroatoms, wherein the heteroatoms have the meaning as described in this invention, wherein each ring system contains 3-7 membered rings and has only one attachment site connected to the rest of the molecule. The term "heteroaryl" may be used interchangeably with the terms "aromatic heterocyclic" or "heteroaromatic compound". Furthermore, the heteroaryl group can be substituted or unsubstituted, wherein the substituent can be, but is not limited to, hydroxyl, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkylamino, alkyl, alkenyl, alkynyl, heterocyclic, mercapto, nitro, aryloxy, hydroxy-substituted alkoxy, hydroxy-substituted alkyl-C(=O)-, alkyl-C(=O)-, alkyl-S(=O)2-, hydroxy-substituted alkyl-S(=O)-, hydroxy-substituted alkyl-S(=O)2-, carboxyalkoxy, etc.

[0086] Other embodiments include, but are not limited to, the following monocyclic compounds: 2-furanyl, 3-furanyl, N-imidazolyl, 2-imidazolyl, 4-imidazolyl, 5-imidazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 4-methylisoxazol-5-yl, N-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 2-pyridyl, 3-pyridyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidinyl, pyrimidin-5-yl, pyridazinyl (e.g., 3-pyridazinyl), 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, tetrazolyl (e.g., 5-tetrazolyl), triazolyl (e.g., 2-triazolyl and 5-triazolyl), 2-thienyl, 3-thienyl, pyrazolyl (e.g., 2-pyrazolyl) ), isothiazolyl, 1,2,3-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,3-thiodiazolyl, 1,3,4-thiodiazolyl, 1,2,5-thiodiazolyl, 1,3,4-thiadiazol-2-yl, pyrazinyl, pyrazin-2-yl, 1,3,5-triazinyl; also includes the following bis Cyclic, but not limited to these bicyclic rings: benzimidazolyl, benzofuranyl, benzothiophenyl, indolyl (e.g., 2-indolyl), purinyl, quinolinyl (e.g., 2-quinolinyl, 3-quinolinyl, 4-quinolinyl), and isoquinolinyl (e.g., 1-isoquinolinyl, 3-isoquinolinyl, or 4-isoquinolinyl), benzo[d]thiazolyl-2-yl, imidazo[1,5-a]pyridin-6-yl.

[0087] The term “heteroatom” refers to one or more O, S, N, P, and Si atoms, including N, S, and P in any oxidation state; in the form of primary, secondary, tertiary amines and quaternary ammonium salts; or in the form where the hydrogen atom on the nitrogen atom in the heterocycle is substituted, for example, N (e.g., N in 3,4-dihydro-2H-pyrrole), NH (e.g., NH in pyrrolealkyl), or NR (e.g., NR in N-substituted pyrrolealkyl).

[0088] The term "halogen" refers to F, Cl, Br, or I.

[0089] In this invention, "halogenated" means replacing the following group with a halogen, and the number of substitutions can be one or more.

[0090] In this invention, "hydroxyl-substituted" means that the group following it is replaced by a hydroxyl group, and the number of substitutions can be one or more.

[0091] When the term "substituted" is used between two groups in this invention, it is preceded by a substituent, such as "aryl-substituted alkyl" indicating that the alkyl group has an aryl substituent, and "alkoxycarbonyl-substituted alkyl" indicating that the alkyl group has an alkoxycarbonyl substituent.

[0092] When multiple groups of the present invention are used in combination, from left to right, they are in a substitution relationship, such as "arylalkyl", which means aryl-substituted alkyl, and "alkoxyalkoxy", which means alkoxy-substituted alkoxy.

[0093] The term "unsaturated" as used in this invention means that a structural portion contains one or more degrees of unsaturation.

[0094] Description of the compounds of the present invention

[0095] In a first aspect, the present invention provides a cyclic peptide compound having a structure as shown in Formula (I), or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of Formula (I).

[0096] Among them, R 1 It can be hydrogen, deuterium, alkyl, haloalkyl, heteroalkyl, or cycloalkyl;

[0097] R 2 It can be hydrogen, deuterium, alkyl, heterocyclic, or heteroaryl;

[0098] R 3 It is aryl or heteroaryl;

[0099] R 4 It is aryl or heteroaryl;

[0100] R 5 It can be amino, alkyl, alkoxy, heteroaryl, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3, -S(=O)-CH3 or -C(=O)-OR 9 ;

[0101] R 6 Aryl, heteroaryl, heterocyclic, or cycloalkyl;

[0102] R 7 and R 8 Each of these can be independently hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, alkenylalkyl, cycloalkylalkyl, haloalkoxy, cycloalkyloxy, heterocyclicoxy, cycloalkyl, heterocyclic, aryl, or heteroaryl.

[0103] Optionally, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles;

[0104] R 9 It can be hydrogen, alkyl, haloalkyl, deuterated alkyl, hydroxyalkyl, or cycloalkyl;

[0105] R 10 It can be hydrogen, deuterium, halogen, hydroxyl, alkyl, haloalkyl, deuterated alkyl, hydroxyalkyl, or cycloalkyl;

[0106] A is -(CH2) p -or-(CH2) q -O-(CH2) u -;

[0107] m, n, t, p, q or u are each independently 0, 1, 2, 3, 4 or 5;

[0108] R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 The groups described in A are independently and optionally replaced by 1, 2, 3 or 4 substituents selected from deuterium, halogen, amino, hydroxyl, cyano, oxo, nitro, alkyl, alkoxy, alkenyl, alkynyl, haloalkyl, cyanoalkyl, alkoxyalkyl, cycloalkylalkyl, heteroalkyl, heteroalkylalkyl, alkenylalkyl, alkynylalkyl, aryl, heteroaryl, heterocyclic, and cycloalkyl.

[0109] In some embodiments, the cyclic peptide compound is described, wherein,

[0110] R 5 For amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-9 heteroaryl, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3, -S(=O)-CH3 or -C(=O)-OR 9 ;

[0111] R 6 C 6-10 Aryl, C 1-9 heteroaryl, C 1-9 Heterocyclic group or C 3-8 cycloalkyl;

[0112] R 9 For hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl or C 3-8 Cycloalkyl.

[0113] In some other embodiments, the cyclic peptide compound, wherein,

[0114] R 5 It is an amino group, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3 or -S(=O)-CH3;

[0115] R 6 C 6-10 Aryl or C 1-9 heteroaryl; preferably

[0116] In some embodiments, the compound has a structure as shown in formula (II), or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of a compound with a structure as shown in formula (II).

[0117] Among them, R 1 R 2 R 3 R 4 R 7 R 8 A, m, and n are defined in equation (I).

[0118] In some embodiments, the cyclic peptide compound, wherein,

[0119] Among them, R 1 For hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Heteroalkyl or C 3-8 cycloalkyl;

[0120] R 2 For hydrogen, deuterium, C 1-6 Alkyl, C 1-9 Heterocyclic groups or independently optionally halogenated, C 1-4 Alkyl-substituted C 1-9 Mixed aromatic compounds.

[0121] In some other embodiments, the cyclic peptide compound, wherein, R 1C is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, or optionally substituted with 1, 2, 3, 4, 5, or 6 F, Cl, Br, or OH groups. 1-4 alkyl;

[0122] R 2 C 1-4 Alkyl group, independently and optionally with 1, 2, or 3 halogens or with C 1-4 Alkyl-substituted pyrrole, pyrazol, imidazole, or triazolyl groups.

[0123] In some embodiments, the cyclic peptide compound, wherein R 3 Or R 4 To be independently and optionally halogenated or C 1-4 Alkyl-substituted C 6-10 Aryl or C 1-9 Mixed aromatic compounds.

[0124] In some other embodiments, the cyclic peptide compound, wherein R 3 To be optionally subjected to 1, 2, 3 F, Cl, Br or C 1-4 Alkyl-substituted pyrrole, pyrazolyl, imidazolyl, triazolyl, phenyl, naphthyl, pyridyl, or C 5-9 Dense heteroaryl compounds.

[0125] In some other embodiments, the cyclic peptide compound, wherein R 4 for

[0126] In some embodiments, the cyclic peptide compound, wherein R 7 and R 8 Each of these can be independently represented by hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, or C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Heteroalkyl, C 1- 6-alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy C 1-6 Alkyl, C 2-6 alkenyl C 1-6 Alkyl, C 3-6 cycloalkyl C 1-6 Alkyl, C 1-6 Halogenated alkoxy groups, C 3-6 Cycloalkyloxy, C 1-6 Heterocyclic oxygen, C 3-6 cycloalkyl, C 1-6 Heterocyclic group, C6-10 Aryl or C 1-6 heteroaryl; optionally, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles;

[0127] R 10 Hydrogen, deuterium, halogen, hydroxyl, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Deuterated alkyl, C 1-3 Hydroxyalkyl or C 3- 6-Cycloalkyl.

[0128] In some other embodiments, the cyclic peptide compound, wherein R 7 and R 8 Each of these can be independently represented by hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, or C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Heteroalkyl, C 1-3 Alkoxy, C 1-3 Hydroxyalkyl or C 1-3 Alkoxy C 1-3 Alkyl; alternatively, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles;

[0129] R 10 For hydrogen, deuterium, halogen, hydroxyl, methyl, ethyl, C substituted with 1, 2, or 3 F or Cl atoms 1-3 Alkyl, C 1- 3-Deuterated alkyl, C 1-3 Hydroxyalkyl or C 3-6 Cycloalkyl.

[0130] In some embodiments, the cyclic peptide compound has one of the following structures, or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug.

[0131] Compositions, formulations and administration of the compounds of the present invention

[0132] The pharmaceutical composition comprises any of the compounds of the present invention. The pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, medium, or combination thereof.

[0133] Substances that can serve as pharmaceutically acceptable carriers include, but are not limited to, ion exchangers, aluminum, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering agents such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-blocking polymers, lanolin, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as carboxymethyl cellulose. Sodium thiosulfate, ethyl cellulose and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic salts; Ringer's solution; ethanol, phosphate buffer solution, and other non-toxic and suitable lubricants such as sodium lauryl sulfate and magnesium stearate, colorants, release agents, coatings, sweeteners, flavorings and spices, preservatives and antioxidants.

[0134] When applicable for treatment, a therapeutically effective amount of the compound of the present invention may be administered as an unprocessed chemical drug or as an active ingredient in a pharmaceutical composition. Therefore, the present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of the compound of the present invention and one or more pharmaceutically acceptable carriers, diluents, or excipients. The term "therapeuticly effective amount" as used herein refers to the total amount of each active component sufficient to demonstrate a meaningful patient benefit (e.g., reduction of viral load). When administered alone using a single active ingredient, the term refers only to that component. When used in combination, the term refers to the combined amount of active ingredients that, regardless of combination, sequential or simultaneous administration, produce a therapeutic effect. The carrier, diluent, or excipient must be acceptable in the sense of compatibility with other components of the formulation and harmlessness to the recipient. According to another aspect of the present invention, a method for preparing a pharmaceutical formulation is also provided, comprising mixing the compound of the present invention with one or more pharmaceutically acceptable carriers, diluents, or excipients. As used in this invention, the term "pharmaceutically acceptable" means that the compounds, raw materials, compositions, and / or dosage forms of this invention are suitable for contact with patient tissues without excessive toxicity, irritation, allergic reactions, or other problems and complications commensurate with a reasonable benefit / risk ratio, and are effective for their intended use, within the limits of reasonable medical judgment.

[0135] It should be understood that, in addition to the ingredients specifically mentioned above, the formulation may also include other ingredients commonly used in the art in relation to the type of formulation, such as flavoring agents, for example, such formulations suitable for oral administration may include flavoring agents.

[0136] Uses of the compounds and compositions of the present invention

[0137] This invention provides the use of the compound or pharmaceutical composition described herein in the preparation of a medicament for the prevention, treatment, relief or regulation of diseases related to or mediated by melanocortin receptors.

[0138] The diseases described in this invention include, but are not limited to, one or more of the following: dry eye syndrome, corneal ulcer, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scarring, epithelial defects, keratoconjunctivitis, idiopathic uveitis, corneal transplantation, age-related macular degeneration, diabetic retinopathy, blepharitis, glaucoma, ocular hypertension, postoperative eye pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascularization, vascular occlusive diseases, and so on. Allergic eye diseases, tumors, retinitis pigmentosa, eye infections, scleritis, ptosis, miosis, eye pain, mydriasis, neuralgia, cicatricial ocular surface diseases, eye infections, inflammatory eye diseases, ocular surface diseases, corneal diseases, retinal diseases, ocular manifestations of systemic diseases, hereditary eye diseases, ocular tumors, increased intraocular pressure, herpes infections, pterygium, wounds extending to the ocular surface, post-refractive keratotomy eye pain and inflammation, corneal thermal or chemical burns, scleral wounds, keratoconus or conjunctival wounds.

[0139] The “effective amount” or “effective dose” of the compounds or pharmaceutically acceptable compositions of the present invention refers to an effective amount for treating or reducing the severity of one or more of the conditions mentioned in the present invention. According to the methods of the present invention, the compounds and compositions thereof can be administered at any dosage and via any route of administration to effectively treat or reduce the severity of the disease. The precise amount required will vary depending on the patient’s condition, which may depend on age, the patient’s general condition, the severity of the infection, specific factors, the route of administration, etc. The compounds or compositions of the present invention may be administered in combination with one or more other therapeutic agents, as discussed in the present invention.

[0140] To enable those skilled in the art to better understand the technical solutions of the present invention, some non-limiting embodiments are further disclosed below to provide a more detailed description of the present invention.

[0141] General Synthesis Process

[0142] Generally, the compounds of the present invention can be prepared by the methods described herein. The following reaction schemes and examples are provided to further illustrate the content of the present invention.

[0143] Those skilled in the art will recognize that the chemical reactions described in this invention can be suitably used to prepare many other compounds of this invention, and that other methods for preparing the compounds of this invention are considered to be within the scope of this invention. For example, the synthesis of those non-illustrative compounds according to this invention can be successfully accomplished by those skilled in the art through modification methods, such as by appropriately protecting interfering groups, by utilizing other known reagents besides those described in this invention, or by making some conventional modifications to the reaction conditions. Furthermore, the reactions disclosed in this invention or the known reaction conditions are also generally accepted to be applicable to the preparation of other compounds of this invention.

[0144] The examples described below, unless otherwise stated, all temperatures are in degrees Celsius. Reagents were purchased from commercial suppliers such as Aldrich Chemical Company, Inc., Arco Chemical Company, and Alfa Chemical Company, and were used without further purification unless otherwise stated. Common reagents were purchased from Shantou Xilong Chemical Plant, Guangdong Guanghua Chemical Reagent Plant, Guangzhou Chemical Reagent Plant, Tianjin Haoyuyu Chemical Co., Ltd., Qingdao Tenglong Chemical Reagent Co., Ltd., and Qingdao Haiyang Chemical Plant.

[0145] Anhydrous tetrahydrofuran, dioxane, toluene, and diethyl ether are obtained by reflux drying with metallic sodium. Anhydrous dichloromethane and chloroform are obtained by reflux drying with calcium hydride. Ethyl acetate, petroleum ether, n-hexane, N,N-dimethylacetamide, and N,N-dimethylformamide are used after prior drying with anhydrous sodium sulfate.

[0146] The following reactions are generally carried out under positive pressure of nitrogen or argon or with a drying tube attached to an anhydrous solvent (unless otherwise specified). All reaction flasks are sealed with suitable rubber stoppers, and the substrate is injected using a syringe. All glassware is dried.

[0147] Silica gel columns were used. Silica gel (300-400 mesh) was purchased from Qingdao Ocean Chemical Plant. Nuclear magnetic resonance spectroscopy used CDCl3, d6-DMSO, CD3OD, or d6-acetone as solvents (reported in ppm), with TMS (0 ppm) or chloroform (7.25 ppm) as reference standards. When multiplets were observed, the following abbreviations were used: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broadened), dd (doublet of doublets), dt (doublet of triplets). Coupling constants were expressed in Hertz (Hz).

[0148] The following abbreviations are used throughout this invention: HOBt: 1-Hydroxybenzotriazole HBTU: O-benzotriazole-tetramethylurea hexafluorophosphate AcOH: Acetic acid Boc2O, BOC acid anhydride: Di-tert-butyl dicarbonate Boc: tert-butyloxycarbonyl Bu4NHSO4: Tetrabutylammonium hydrogen sulfate CH3CN, ACN: Acetonitrile DCM: Dichloromethane DIPEA: N,N-diisopropylethylamine EA: Ethyl acetate HCl: Hydrogen chloride HCl / EA: Ethyl acetate solution of hydrogen chloride H2O: Water NaOH: Sodium hydroxide NaI: Sodium iodide K2CO3: Potassium carbonate rt, rt: Room temperature PE: Petroleum ether THF: Tetrahydrofuran EDCI: 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride DMAP: 4-Dimethylaminopyridine TBAI: Tetrabutylammonium iodide DMF: Dimethylformamide p-TsOH: p-Toluenesulfonic acid TFA: Trifluoroacetic acid TBAF: Tetrabutylammonium fluoride FA: Formic acid CDI: N,N-carbonyldiimide

[0149] Example

[0150] Intermediate 1

[0151] Step 1: Synthesis of 5-aminohexanoic acid (int-1b)

[0152] In a 500 mL round-bottom flask, compound int-1a (25 g, 224.24 mmol) was dissolved in 6 M HCl (200 mL), and the mixture was refluxed for 24 h. The reaction was monitored by LC-MS until completion. After cooling to room temperature, the solvent was removed under reduced pressure to obtain compound int-1b, a white solid (28.1 g, yield: 97.1%). The crude product was used directly in the next step without purification. LC-MS: 132.1 [M+1] + .

[0153] Step 2: Synthesis of 5-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)hexanoic acid (int-1c)

[0154] In a 500 mL round-bottom flask, compound int-1b (15 g, 114.5 mmol) and sodium carbonate (24.3 g, 229.0 mmol) were dissolved in tetrahydrofuran / water (100 mL / 100 mL). A tetrahydrofuran solution of 9-fluorenylmethyl-N-succinimide carbonate (90.8 g, 269.09 mmol) was added dropwise under ice bath conditions. After the addition was complete, the reaction was allowed to proceed at room temperature for 3 h. The reaction was monitored by TLC until completion. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The residue was purified by column chromatography (PE:EA = 5:1) to give compound int-1c, a white solid (20.8 g, yield: 50.7%). LCMS: 354.4 [M+1] + Step 3: Synthesis of (9H-fluorene-9-yl)methyl tert-butylpentane-1,4-dimethyl dicarboxylate (int-1d)

[0155] In a 500 mL round-bottom flask, compound int-1c (18.0 g, 50.99 mmol), triethylamine (6.7 g, 66.29 mmol), and tert-butanol (200 mL) were added dropwise. Diphenyl azidophosphate (18.3 g, 66.29 mmol) was added dropwise under ice bath conditions. The reaction was carried out at room temperature for 3 h. TLC monitoring showed the starting material had disappeared, and the mixture was then refluxed for 6 h. LCMS monitoring indicated the reaction was complete. The mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 8:1) to give compound int-1d, a white solid (7.8 g, yield: 35.4%). LCMS: 425.2 [M+1] + .

[0156] Step 4: Synthesis of (9H-fluorene-9-yl)methyl(5-aminopent-2-yl)carbamate (int-1e)

[0157] In a 100 mL round-bottom flask, compound int-1d (3.4 g, 8.0 mmol) was dissolved in dichloromethane (30 mL). The mixture was cooled to 0 °C in an ice bath, and 10 mL of 4 M dioxane hydrochloride solution was slowly added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 2 h. The reaction was monitored by LCMS until completion. The organic solvent was removed under reduced pressure to obtain compound int-1e, a white solid (2.67 g), which was used directly in the next step without purification. LCMS: 325.2 [M+1] + .

[0158] Step 5:

[0159] In a 250 mL solid-phase peptide synthesis tube, 5 g of 2-chlorotriphenylmethyl resin (2-CTC resin) with a substitution degree of 1.2-1.4 mmol / g was added. After swelling with DMF (50 mL) for 30 min, compound int-1e (2.6 g, 8.0 mmol) was dissolved in DMF (20 mL). After activation with DIPEA (2.1 g, 16 mmol) at 0 °C, the solution was added to the resin-packed reaction column. After reacting for 2 hours, the mixture was washed three times with DMF and three times with DCM, and the solvent was dried. 20 mL of 20% morpholine / DMF solution was added, and the reaction was continued for 30 min. The mixture was then washed three times with DMF and three times with DCM, and the solvent was dried to obtain compound int-1f. LCMS: 389.2 [M+1] + .

[0160] Step 6:

[0161] N-(9-fluorenylmethoxycarbonyl)-N1-tert-butyloxycarbonyl-L-tryptophan (8.7 g, 16.50 mmol) and 1-hydroxybenzotriazole (2.23 g, 16.50 mmol) were dissolved in 60 mL of a 1:1 mixture of DCM and DMF. Benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (6.25 g, 16.50 mmol) was added at 0 °C and activated for 10 min. The solution was then added to the solid-phase reaction column from the previous step and reacted at room temperature for 30 min. The mixture was washed three times with DMF and three times with DCM, and the solvent was removed under vacuum. 20 mL of 20% morpholine / DMF solution was added, and the reaction was continued for another 30 min. The mixture was then washed three times with DMF and three times with DCM, and the solvent was removed under vacuum to obtain compound int-1 g. LCMS: 797.4 [M+1] + .

[0162] Step 7:

[0163] N-fluorenylmethoxycarbonyl-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl-L-arginine (10.7 g, 16.50 mmol) and HOBt (2.23 g, 16.50 mmol) were dissolved in 60 mL of a 1:1 mixture of DCM and DMF. After activation for 10 min at 0 °C, HBTU (6.25 g, 16.50 mmol) was added to the solid-phase reaction column from the previous step. The reaction was carried out at room temperature for 30 min, followed by washing three times with DMF and three times with DCM, and the solvent was removed under vacuum. 20 mL of 20% morpholine / DMF solution was added, and the reaction was continued for another 30 min. The mixture was then washed three times with DMF and three times with DCM, and the solvent was removed under vacuum to obtain compound int-1h. LCMS: 797.4 [M+1] + .

[0164] Step 8:

[0165] Dissolve (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid (6.7 g, 16.50 mmol) and HOBt (2.23 g, 16.50 mmol) in 60 mL of a 1:1 mixture of DCM and DMF. Activate the solution by adding HBTU (6.25 g, 16.50 mmol) at 0 °C for 10 min, then add the solution to a solid-phase reaction column. React at room temperature for 30 min, then wash three times with DMF and three times with DCM, and remove the solvent. Add 20 mL of 20% morpholine / DMF solution, continue the reaction for 30 min, then wash three times with DMF and three times with DCM, and remove the solvent to obtain intermediate 1, a pale yellow solid (12.3 g). LCMS: 948.5 [M+1] + .

[0166] Intermediate 2a and intermediate 2b

[0167] Step 1: (9H-fluorene-9-yl)methyl tert-butylpentane-1,4-diyl(R)-dicarbamate and (9H-fluorene-9-yl)methyl tert-butylpentane-1,4-diyl(S)-dicarbamate tert-butyl ester (int-2a and int-2b)

[0168] Compound int-1e (6.6 g, 15.49 mmol) was chirally resolved to yield compounds int-2a (3.4 g, yield: 51.5%, chiral HPLC RT = 6.754 min) and int-2b (3.0 g, yield: 45.5%, chiral HPLC RT = 6.899 min). LCMS: 425.2 [M+1] + .

[0169] Step 2: (9H-fluorene-9-yl)methyl(R)-(5-aminopent-2-yl)carbamate (int-2c)

[0170] In a 100 mL round-bottom flask, compound int-2a (3.0 g, 7.1 mmol) was dissolved in dichloromethane (30 mL). The mixture was cooled to 0 °C in an ice bath, and 10 mL of 4 M dioxane hydrochloride solution was slowly added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 2 h. The reaction was monitored by LCMS until completion. The organic solvent was removed under reduced pressure to obtain compound int-2c, a white solid (2.4 g), which was used directly as a pre-purified sample for the next step. LCMS: 325.2 [M+1] + .

[0171] Step 3: (9H-fluorene-9-yl)methyl(S)-(5-aminopent-2-yl)carbamate (int-2d)

[0172] In a 100 mL round-bottom flask, compound int-2b (3.0 g, 7.1 mmol) was dissolved in dichloromethane (30 mL). The mixture was cooled to 0 °C in an ice bath, and 10 mL of 4 M dioxane hydrochloride solution was slowly added dropwise. After the addition was complete, the reaction was allowed to proceed at room temperature for 2 h. The reaction was monitored by LCMS until completion. The organic solvent was removed under reduced pressure to obtain compound int-2d, a white solid (2.2 g), which was used directly in the next step without purification. LCMS: 325.2 [M+1] + .

[0173] Step 4:

[0174] In a 250 mL solid-phase polypeptide synthesis tube, 5.5 g of 2-CTC resin with a substitution degree of 1.2-1.4 mmol / g was added and swollen with DMF (50 mL) for 30 min. Then, 2.6 g (8.0 mmol) of compound int-2c was dissolved in DMF (20 mL), activated with DIPEA (2.1 g, 16 mmol) at 0 °C, and added to the above-mentioned reaction column containing resin. After reacting for 2 hours, the mixture was washed 3 times with DMF and 3 times with DCM, and the solvent was dried. 20 mL of 20% morpholine / DMF solution was added, and the reaction was continued for 30 min. The mixture was then washed 3 times with DMF and 3 times with DCM. Subsequently, O-benzotriazole-tetramethylurea hexafluorophosphate and N-fluorenylmethoxycarbonyl-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl-L-arginine were condensed and deprotected to obtain intermediate 2a. LCMS: 797.4 [M+1] + .

[0175] Step 5:

[0176] In a 250 mL solid-phase peptide synthesis tube, 4.5 g of 2-CTC resin with a substitution degree of 1.2-1.4 mmol / g was added. After swelling with DMF (50 mL) for 30 min, compound int-2d (2.1 g, 6.48 mmol) was dissolved in DMF (20 mL). After activation with DIPEA (1.67 g, 12.96 mmol) at 0 °C, the solution was added to the reaction column containing the resin. After reacting for 2 hours, the solution was analyzed using D... Wash three times with MF and three times with DCM, then remove the solvent by vacuum. Add 20% morpholine / DMF solution (20 mL), continue the reaction for 30 min, then wash three times with DMF and three times with DCM. Subsequently, condensate and deprotect N-(9-fluorenylmethoxycarbonyl)-N1-tert-butoxycarbonyl-L-tryptophan and N-fluorenylmethoxycarbonyl-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl-L-arginine sequentially using the int-1g method to obtain intermediate 2b. LCMS: 797.4 [M+1] + .

[0177] Intermediate 3

[0178] first step:

[0179] In a 250 mL solid-phase peptide synthesis tube, 15 g of 2-CTC resin with a substitution degree of 1.2-1.4 mmol / g was added. After swelling with DMF (80 mL) for 30 min, compound int-3a (6.7 g, 21.6 mmol) was dissolved in DMF (80 mL), activated with DIPEA (5.57 g, 43.2 mmol) at 0 °C, and then added to the above-mentioned reaction column packed with resin. After reacting for 2 hours, the solution was transferred to a DMF-containing column. Wash three times with F and three times with DCM, then remove the solvent by vacuum. Add 80 mL of 20% morpholine / DMF solution and continue the reaction for 30 min. Wash three times with DMF and three times with DCM. Then, condensate and deprotect N-(9-fluorenylmethoxycarbonyl)-N1-tert-butoxycarbonyl-L-tryptophan and N-fluorenylmethoxycarbonyl-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl-L-arginine sequentially to give compound int-3b (23 g). LCMS: 797.4 [M+1] + .

[0180] Step Two:

[0181] In a 500 mL solid-phase polypeptide synthesis tube, compound int-3b (15 g) was added, followed by swelling with DMF (100 mL) for 30 min. Then, (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid (18.3 g, 45.0 mmol) and HOBt (6.1 g, 45.0 mmol) were dissolved in 80 mL of a 1:1 mixture of DCM and DMF. After activation at 0 °C for 10 min, HBTU (17.1 g, 45.0 mmol) was added to the solid-phase reaction column. The reaction was carried out at room temperature for 30 min, followed by washing three times with DMF and three times with DCM, and the solvent was removed. A 20% morpholine / DMF solution (100 mL) was added, and the reaction was continued for 30 min. The mixture was then washed three times with DMF and three times with DCM, and the solvent was removed to obtain intermediate 3. LCMS: 948.5 [M+1] + .

[0182] Example 1

[0183] (S)-N-((2S,5R,8S,11S,22S)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-2-((1-methyl-1H-imidazol-4-yl)methyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic dicyclic sugar-22-yl)-2-acetaminohexanamide (Compound 1)

[0184] Step 1: Coupling and Deprotection of Amino Acids

[0185] Following the synthetic method of compound int-1g, intermediate 3 (1.2 g) was added to a polypeptide synthesis reaction tube, followed by condensation and deprotection of N-[fluorenylmethoxycarbonyl]-1-methyl-L-histidine (1.41 g, 3.6 mmol), N-[(9H-fluoren-9-ylmethoxy)carbonyl]-L-glutamic acid 5-tert-butyl ester (1.53 g, 3.6 mmol), and fluorenylmethoxycarbonyl acyl leucine (1.27 g, 3.6 mmol) to obtain compound 1a. LCMS: 1625.8 [M+1] + .

[0186] Step 2: Acetylation

[0187] In a peptide synthesis reaction tube, compound 1a (1.44 mmol), pyridine (171 mg, 2.16 mmol), acetic anhydride (177 mg, 1.73 mmol), and DMF (10 mL) were added. After reacting at room temperature for 1 h, the mixture was washed three times with DMF and three times with DCM. The solvent was then removed by vacuum to obtain compound 1b (2.4 g). LCMS: 1667.8 [M+1] + .

[0188] Step 3: Cutting and removing protection

[0189] In a 100 mL round-bottom flask, compound 1b (2.4 g), triisopropylsilane (0.5 mL), water (0.5 mL), and trifluoroacetic acid (10 mL) were added sequentially, and the mixture was reacted at room temperature for 3 h. The reaction was monitored by LCMS until completion. The mixture was filtered, and the filtrate was concentrated by passing it through a nitrogen stream. The residue was added to methyl ether (10 mL), and the mixture was left to stand overnight at 0 °C. Filtering yielded compound 1c, a white solid (861 mg, yield: 58.1%). LCMS: 1031.6 [M+1] + .

[0190] Step 4: Cycloning

[0191] (S)-N-((2S,5R,8S,11S,22S)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-2-((1-methyl-1H-imidazol-4-yl)methyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic dicyclic sugar-22-yl)-2-acetaminohexanamide (Compound 1)

[0192] In a 100 mL round-bottom flask, compound 1c (860 mg, 0.834 mmol) was dissolved in DMF / THF (3 / 7 mL). The mixture was cooled to 0 °C in an ice bath, and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (434 mg, 0.834 mmol) was added. After reacting at this temperature for 10 min, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (868 mg, 1.67 mmol) and N-methylmorpholine (843 mg, 8.26 mmol) were added, and the mixture was slowly heated to room temperature for 3 h. The reaction was monitored by LCMS until completion. The reaction was quenched with 1 mL of dilute hydrochloric acid (1 M), and the organic solvent was removed under reduced pressure. The residue was slurried with ethyl acetate and tertiary methyl ether (1:1) and filtered to obtain the crude product. The crude product was purified by preparative reversed-phase high-performance chromatography (RP-HPLC) to obtain compound 1, a white solid (110 mg, yield: 13%). 1 H NMR(400MHz,DMSO-d6)δ10.81(s,1H),8.51(d,1H),8.33(d,1H),8.28(s,1H),8.17(d,1H),8.02(t,3H),7.79(d,2H),7.59(t,2H),7.41(s,1H), 7.34(d,1H),7.22(dd,2H),7.13(d,1H),7.09(d,1H),7.07(s,2H),7.05 (s,1H),6.98(t,1H),6.66(s,1H),4.49–4.39(m,2H),4.36–4.28(m,2H), 4.25–4.15(m,2H),3.53(s,3H),3.24(d,2H),3.19-3.13(m,2H),3.05(d ,2H),3.01(d,3H),2.88–2.75(m,2H),2.71–2.56(m,2H),2.06(s,2H),1. 90(d,1H),1.85(s,3H),1.82–1.73(m,1H),1.66(dd,2H),1.53–1.42(m, 2H),1.40–1.30(m,5H),1.24(d,4H),0.85(d,3H)ppm; LCMS: 1013.5[M+1] + .

[0193] Example 2

[0194] (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetamido-5-(4H-1,2,4-triazol-4-yl)pentanamide (Compound 2)

[0195] Step 1: (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-hydroxyvalerate tert-butyl ester (2b)

[0196] In a 500 mL round-bottom flask, compound 2a (10.0 g, 23.52 mmol) was dissolved in 200 mL of THF. 1-Hydroxybenzotriazole (4.76 g, 35.27 mmol) was added, followed by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (6.76 g, 35.27 mmol). The mixture was stirred at room temperature for 2 h, and the reaction was stopped by LCMS. 20 mL of water was added, and the mixture was cooled to 0 °C in an ice bath. Sodium borohydride (1.79 g, 47.04 mmol) was then added, and the reaction was stopped by LCMS. The reaction was quenched with 100 mL of saturated ammonium chloride solution. The organic solvent was removed under reduced pressure. The residue was extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The crude product was purified by silica gel column chromatography to give compound 2b, an anhydrous oil (7.5 g, yield: 78%). LCMS: 412.2 [M+1] + Step 2: (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-(4H-1,2,4-triazol-4-yl)tert-butyl valerate (2c)

[0197] In a 500 mL round-bottom flask, compound 2b (7.5 g, 18.25 mmol) was dissolved in 100 mL of THF. The mixture was cooled to 0 °C in an ice bath, and triphenylphosphine (7.2 g, 27.36 mmol) was added. Then, diisopropyl azodicarbonate (5.53 g, 27.36 mmol) was slowly added dropwise. The mixture was brought to room temperature and stirred for 2 h. The reaction was stopped by LCMS. 20 mL of water was added, and the mixture was cooled to 0 °C in an ice bath. Sodium borohydride (1.79 g, 47.04 mmol) was then added. The reaction was stopped by LCMS. The organic solvent was removed under reduced pressure, and the crude product was purified by silica gel column chromatography to give compound 2c, an anhydrous oil (5.5 g, yield: 65%). LCMS: 462.2 [M+1] + Step 3: (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-(4H-1,2,4-triazol-4-yl)valeric acid (2d)

[0198] In a 250 mL round-bottom flask, compound 2c (5.5 g, 11.90 mmol) was dissolved in 50 mL of DCM, followed by the addition of 20 mL of 4 M HCl solution and 4-dioxane solution. The mixture was stirred at room temperature for 2 h, and the reaction was monitored by LCMS until completion. The organic solvent was removed under reduced pressure to give compound 2d, a white solid (4.7 g, yield: 96%). LCMS: 407.1 [M+1] + .

[0199] Step 4: (7S,10S,13R,16S,19S)-16-((1H-imidazol-4-yl)methyl)-7-((1H-indol-3-yl)methyl)-19-(((S)-2-acetamido-5-(4H-1,2,4-triazol-4-yl)pentamido)-1-amino-13-(4-fluorobenzyl)-10-(3-guanidinopropyl)-6,9,12,15,18-pentoxo-5,8,11,14,17-pentazaeicosazacyclopenta-22-acid (2e)

[0200] Following the amino acid condensation and deprotection method described in Example 1, starting with intermediate 3 (1.2 g), N-Fmoc-N'-triphenylmethyl-L-histidine, Fmoc-O-tert-butyl-L-glutamic acid, and compound 2d were sequentially condensed, deprotected, acetylated, and cleaved to obtain compound 2e, a white solid (430 mg, yield: 27.9%). LCMS: 1070.5 [M+1] + .

[0201] Step 5: (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetamido-5-(4H-1,2,4-triazol-4-yl)pentanamide (Compound 2)

[0202] In a 100 mL round-bottom flask, compound 2e (430 mg, 0.402 mmol) was dissolved in DMF / THF (3 / 7 mL). The mixture was cooled to 0 °C in an ice bath, and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (209 mg, 0.402 mmol) was added. After reacting at this temperature for 10 min, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (418 mg, 0.804 mmol) and N-methylmorpholine (402 mg, 3.98 mmol) were added, and the mixture was slowly heated to room temperature for 3 h. The reaction was monitored by LCMS until completion. The reaction was quenched with 1 mL of dilute hydrochloric acid (1 M), and the organic solvent was removed under reduced pressure. The residue was slurried with ethyl acetate and tertiary methyl ether (1:1) and filtered to obtain the crude product. The crude product was purified by preparative reversed-phase high-performance chromatography (RP-HPLC) to obtain compound 2, a white solid (31 mg, yield: 7.3%). 1 H NMR(500MHz,DMSO-d6)δ10.79(s,1H),8.46(s,2H),8.30(s,2H),8.22(d,1H), 8.12(d,1H),8.05(d,2H),7.93(s,2H),7.78(s,1H),7.61–7.53(m,2H),7.49( s,1H),7.32(d,2H),7.19(dd,2H),7.12(s,1H),7.07(d,2H),7.05(d,1H),6.9 6(t,1H),6.66(s,1H),4.50–4.40(m,2H),4.38–4.26(m,3H),4.20–4.16(m,1H) ,4.14(t,2H),3.25–3.19(m,3H),3.18–3.10(m,3H),3.04–2.99(m,3H),2.99( s,3H),2.88–2.79(m,2H),2.77–2.70(m,2H),2.63(d,1H),2.08–2.01(m,2H),1 .88(d,1H),1.85–1.81(m,3H),1.80–1.70(m,3H),1.69–1.62(m,1H),1.59(d, 1H),1.43(d,2H),1.38–1.33(m,3H),1.32–1.27(m,2H)ppm; LCMS: 1052.5[M+1] + .

[0203] Example 3

[0204] (S)-N-((2S,5S,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((2,3-dihydrobenzofuran-5-yl)methyl)-8-(3-guanidinopropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic disaccharide-22-yl)-2-acetaminohexanamide and (S)-N -((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((2,3-dihydrobenzofuran-5-yl)methyl)-8-(3-guanidinopropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic disaccharide-22-yl)-2-acetaminohexanamide (compounds 3-1 and 3-2)

[0205] Step 1: (E)-2-((tert-Butoxycarbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)methyl acrylate (3b)

[0206] In a 250 mL round-bottom flask, compound 3a (7.0 g, 47.25 mmol) and N-Boc-2-(dimethylphosphono)glycine methyl ester (18.3 g, 61.42 mmol) were dissolved in dichloromethane (70 mL). 1,8-diazabicyclo[5.4.0]undec-7-ene (9 mL, 61.42 mmol) was added dropwise with stirring. The reaction was allowed to proceed at room temperature for 3 h after the addition was complete. The reaction was monitored by TLC until completion. The organic solvent was removed under reduced pressure, and the residue was purified by column chromatography (PE:EA = 8:1) to give compound 3b, a colorless oily solid (13.6 g, yield: 90.7%). LCMS: 320.1 [M+1] + .

[0207] Step 2: Methyl 2-(tert-Butoxycarbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)propionate (3c)

[0208] Under a hydrogen atmosphere, compound 3b (13.5 g, 42.32 mmol) and palladium on carbon (2.7 g, 10% palladium) were dissolved in anhydrous ethanol (130 mL) in a 250 mL round-bottom flask and reacted overnight at room temperature. The reaction was monitored by LCMS until completion. Palladium on carbon was removed by filtration, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 8:1) to give compound 3c, a colorless oily solid (13 g, yield: 96.2%). LCMS: 322.1 [M+1] + .

[0209] Step 3: Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)propionate and methyl (R)-2-((tert-Butoxycarbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)propionate (3d and 3e)

[0210] Compound 3c (6.9 g, 21.5 mmol) was chirally resolved to give compound 3d (3.3 g, chiral HPLC RT = 8.878 min, ee = 98.602%, yield: 47.8%) and compound 3e (3.4 g, chiral HPLC RT = 9.649 min, ee = 98.834%, yield: 49.2%). LCMS: 322.1 [M+1] + .

[0211] Step 4: (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)propionate (3f)

[0212] In a 100 mL round-bottom flask, compound 3d (3.2 g, 9.97 mmol) was dissolved in dichloromethane (30 mL). Dioxane hydrochloride solution (10 mL, 4 M) was added dropwise under ice bath, and the reaction was carried out at room temperature for 1 h. After TLC monitoring showed the reaction was complete, the solvent was removed under reduced pressure. The residue was dissolved in a mixture of tetrahydrofuran and water (15 / 15 mL). Sodium carbonate (3.2 g, 29.91 mmol) and 9-fluorenylmethyl-N-succinimide carbonate (4.03 g, 11.96 mmol) were added under ice bath, and the reaction was carried out at room temperature for 3 h. After TLC monitoring showed the reaction was complete, the mixture was extracted with ethyl acetate, dried over anhydrous sulfuric acid, filtered, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 4:1-1:1) to give compound 3f, a white solid (2.84 g, yield: 64.2%). LCMS: 444.1 [M+1] + .

[0213] Step 5: (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)propionate (3g)

[0214] In a 100 mL round-bottom flask, compound 3e (3.3 g, 10.28 mmol) was dissolved in dichloromethane (30 mL). Dioxane hydrochloride solution (10 mL, 4 M) was added dropwise under ice bath, and the reaction was carried out at room temperature for 1 h. After TLC monitoring showed the reaction was complete, the solvent was removed under reduced pressure. The residue was dissolved in a mixture of tetrahydrofuran and water (15 / 15 mL). Sodium carbonate (3.3 g, 30.84 mmol) and 9-fluorenylmethyl-N-succinimide carbonate (4.16 g, 12.34 mmol) were added under ice bath, and the reaction was carried out at room temperature for 3 h. After TLC monitoring showed the reaction was complete, the mixture was extracted with ethyl acetate, dried over anhydrous sulfuric acid, filtered, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 4:1-1:1) to give 3 g of compound as a white solid (2.76 g, yield: 60.6%). LCMS: 444.1 [M+1] + .

[0215] Step 6: (S)-2-((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)propionic acid (3h)

[0216] In a 100 mL round-bottom flask, compound 3f (2.8 g, 6.32 mmol), trimethyltin hydroxide (3.41 g, 18.96 mmol), and 1,2-dichloroethane (30 mL) were added sequentially, and the mixture was refluxed for 6 h. After the reaction was complete as monitored by TLC, the mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 3f as a white solid (2.49 g, chiral HPLC RT = 9.876 min, ee = 100%, yield: 91.8%). LCMS: 430.1 [M+1] + .

[0217] Step 7: (R)-2-((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2,3-dihydrobenzofuran-5-yl)propionic acid (3i)

[0218] In a 100 mL round-bottom flask, 3 g (2.7 g, 6.09 mmol) of compound, 3.29 g (18.28 mmol) of trimethyltin hydroxide, and 30 mL of 1,2-dichloroethane were added sequentially, and the mixture was refluxed for 6 h. After the reaction was complete as monitored by TLC, the mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 3i, a white solid (2.4 g, chiral HPLC RT = 9.015 min, ee = 100%, yield: 92.3%). LCMS: 430.1 [M+1] + .

[0219] Step 8: (S)-N-((2S,5S,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((2,3-dihydrobenzofuran-5-yl)methyl)-8-(3-guanidinopropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazabiose-22-yl)-2-acetaminohexanamide (compound 3-1)

[0220] The synthesis of compound 3-1 was performed following the steps in Example 1, using compound int-3b (2.5 g) as the starting material, and replacing the third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid with compound 3h. Compound 3-1 was obtained as a white solid (115 mg, yield: 3.7%).

[0221] 1 H NMR(500MHz,DMSO-d6)δ10.78(s,1H),8.56(s,1H),8.36(d,1H),8.15(s,1H),8.01(d,1H),7.95(d,1H),7.91(s,1H),7.76(s,1H),7.55(d,2H), 7.52(d,2H),7.30(d,2H),7.16(s,1H),7.06(s,1H),7.02(dd,2H),6.99 –6.94(m,1H),6.92(s,1H),6.68(d,1H),4.54–4.40(m,4H),4.31(dd,1H ),4.26–4.14(m,2H),4.00(d,1H),3.29–3.22(m,4H),3.18–3.10(m,4H) ,3.02(d,4H),2.98–2.88(m,3H),2.82(d,2H),2.74–2.62(m,2H),2.22– 2.12(m,1H),2.07(dd,1H),1.86(s,4H),1.62(s,1H),1.55(s,2H),1.42(s,3H),1.35(s,3H),1.21(t,4H),0.83(dd,3H)ppm; LCMS: 1023.5[M+1] + .

[0222] Step 9: (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((2,3-dihydrobenzofuran-5-yl)methyl)-8-(3-guanidinopropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazabiose-22-yl)-2-acetaminohexanamide (compound 3-2)

[0223] The synthesis of compound 3-2 was performed following the steps in Example 1, using compound int-3b (2.5 g) as the starting material, and replacing the third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid with compound 3i. Compound 3-2 was obtained as a white solid (235 mg, yield: 7.7%). 1 H NMR(500MHz,DMSO-d6)δ10.77(s,1H),8.41(d,1H),8.20(s,2H),8.14(s,1H ),8.01(d,1H),7.97(d,1H),7.88(d,1H),7.82(s,1H),7.56(t,2H),7.42(s, 1H),7.33(d,1H),7.12(s,1H),7.08–7.01(m,2H),6.98(t,1H),6.88(d,1H) ,6.81(s,1H),6.63(d,1H),4.55(d,1H),4.51–4.41(m,3H),4.37(s,1H),4.2 6(d,1H),4.23–4.14(m,2H),3.27–3.18(m,4H),3.16–3.04(m,5H),2.99(d, 4H),2.93–2.86(m,1H),2.82(s,2H),2.74–2.63(m,2H),2.37(s,1H),2.04(d ,2H),1.86(s,1H),1.84(s,3H),1.80–1.72(m,1H),1.66(s,1H),1.59(s,1H) ,1.44(s,2H),1.32(d,6H),1.24(s,5H),0.83(d,3H)ppm; LCMS: 1023.5[M+1] + .

[0224] Example 4

[0225] (S)-N-((2S,5S,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((3-fluoropyridin-4-yl)methyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide and (S)-N -((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((3-fluoropyridin-4-yl)methyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (compounds 4-1 and 4-2)

[0226] Step 1: (E)-2-((tert-Butoxycarbonyl)amino)-3-(3-fluoropyridin-4-yl)methyl acrylate (4b)

[0227] In a 100 mL round-bottom flask, compound 4a (3.3 g, 25.88 mmol) and N-Boc-2-(dimethylphosphono)glycine methyl ester (10.0 g, 33.64 mmol) were dissolved in dichloromethane (30 mL). 1,8-diazabicyclo[5.4.0]undec-7-ene (4.9 mL, 33.64 mmol) was added dropwise with stirring. After the addition was complete, the reaction was allowed to proceed at room temperature for 3 h. The reaction was monitored by TLC until completion. The organic solvent was removed under reduced pressure, and the residue was purified by column chromatography (PE:EA = 8:1) to give compound 4b as a white solid (7.35 g, yield: 95.8%). LCMS: 297.1 [M+1] + .

[0228] Step 2: Methyl 2-((tert-Butoxycarbonyl)amino)-3-(3-fluoropyridin-4-yl)propionate (4c)

[0229] Under a hydrogen atmosphere, compound 4b (7.3 g, 24.66 mmol) and palladium on carbon (1.46 g, 10% palladium) were dissolved in anhydrous ethanol (70 mL) in a 250 mL round-bottom flask and reacted overnight at room temperature. The reaction was monitored by LCMS until completion. Palladium on carbon was removed by filtration, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 8:1) to give compound 4c, a white solid (7.21 g, yield: 98.7%). LCMS: 299.1 [M+1] + .

[0230] Step 3: Methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoropyridin-4-yl)propionate and methyl (R)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoropyridin-4-yl)propionate (4d and 4e)

[0231] Compound 4c (6.8 g, 22.82 mmol) was chirally resolved to give compound 4d (3.1 g, chiral HPLC RT = 7.758 min, ee = 98.949%, yield: 45.6%) and compound 4e (3.0 g, chiral HPLC RT = 8.244 min, ee = 98.945%, yield: 44.1%). LCMS: 299.1 [M+1] + .

[0232] Step 4: (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-fluoropyridin-4-yl)propionate (4f)

[0233] In a 100 mL round-bottom flask, compound 4d (3.0 g, 10.03 mmol) was dissolved in dichloromethane (30 mL), and dioxane hydrochloride solution (10 mL, 4 M) was added dropwise under ice bath. The reaction was carried out at room temperature for 1 h. After TLC monitoring showed the reaction was complete, the solvent was removed under reduced pressure. The residue was dissolved in a mixture of tetrahydrofuran and water (15 / 15 mL), and sodium carbonate (3.2 g, 30.1 mmol) and 9-fluorenylmethyl-N-succinimide carbonate (4.06 g, 12.04 mmol) were added under ice bath. The reaction was carried out at room temperature for 3 h. After TLC monitoring showed the reaction was complete, the mixture was extracted with ethyl acetate, dried over anhydrous sulfuric acid, filtered, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 4:1-1:1) to give compound 4f, a white solid (2.87 g, yield: 68.1%). LCMS: 421.1 [M+1] + Step 5: (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-fluoropyridin-4-yl)propionate methyl ester (4g)

[0234] In a 100 mL round-bottom flask, compound 4e (2.9 g, 9.73 mmol) was dissolved in dichloromethane (30 mL). Dioxane hydrochloride solution (10 mL, 4 M) was added dropwise under ice bath, and the reaction was carried out at room temperature for 1 h. After TLC monitoring showed the reaction was complete, the solvent was removed under reduced pressure. The residue was dissolved in a mixture of tetrahydrofuran and water (15 / 15 mL). Sodium carbonate (3.1 g, 29.19 mmol) and 9-fluorenylmethyl-N-succinimide carbonate (3.93 g, 11.68 mmol) were added under ice bath, and the reaction was carried out at room temperature for 3 h. After TLC monitoring showed the reaction was complete, the mixture was extracted with ethyl acetate, dried over anhydrous sulfuric acid, filtered, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 4:1-1:1) to give 4 g of compound as a white solid (2.69 g, yield: 65.7%). LCMS: 421.1 [M+1] + Step 6: (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-fluoropyridin-4-yl)propionic acid (4h)

[0235] In a 100 mL round-bottom flask, compound 4f (2.8 g, 6.67 mmol), trimethyltin hydroxide (3.6 g, 20.0 mmol), and 1,2-dichloroethane (30 mL) were added sequentially, and the mixture was refluxed for 6 h. After the reaction was complete as monitored by TLC, the mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 4f as a white solid (2.61 g, chiral HPLC RT = 9.320 min, ee = 100%, yield: 99.2%). LCMS: 407.1 [M+1] + .

[0236] Step 7: (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-fluoropyridin-4-yl)propionic acid (4i)

[0237] In a 100 mL round-bottom flask, 4 g (2.6 g, 6.19 mmol) of compound, 3.34 g (18.57 mmol) of trimethyltin hydroxide, and 30 mL of 1,2-dichloroethane were added sequentially, and the mixture was refluxed for 6 h. After the reaction was complete as monitored by TLC, the mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 4i as a white solid (2.48 g, chiral HPLC RT = 7.493 min, ee = 100%, yield: 98.8%). LCMS: 407.1 [M+1] + .

[0238] Step 8: (S)-N-((2S,5S,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((3-fluoropyridin-4-yl)methyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 4-1)

[0239] The synthesis of compound 4-1 followed the steps in Example 1, using compound int-3b (2.5 g) as the starting material, and replacing the third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid with compound 4h. Compound 4-1 was a white solid (161 mg, yield: 5.3%). 1 H NMR(500MHz,DMSO-d6)δ14.16(s,1H),10.78(s,1H),8.92(s,1H),8.49(s,1H),8.39(s,1H),8.37 (d,2H),8.21(d,1H),8.03(d,1H),7.93(d,1H),7.87–7.81(m,1H),7.71(d,1H),7.58–7.53(m,1H ),7.52(d,2H),7.41–7.36(m,1H),7.27(d,1H),7.24(s,1H),7.08(d,1H),7.03(t,1H),6.96(t,1 H),4.72(s,1H),4.49–4.41(m,1H),4.37–4.30(m,1H),4.21–4.10(m,2H),4.07–3.97(m,1H),3.46 –3.40(m,3H),3.36–3.23(m,2H),3.23–3.18(m,1H),3.17–3.11(m,1H),3.05(s,2H),3.01-2.96( m,1H),2.94(d,1H),2.91(s,1H),2.90–2.86(m,1H),2.84-2.76(m,1H),2.66–2.57(m,1H),2.23–2 .13(m,1H),2.11–2.02(m,1H),1.93–1.87(m,1H),1.85(s,3H),1.83–1.75(m,1H),1.66–1.54(m, 2H),1.53–1.46(m,1H),1.46–1.28(m,7H),1.27–1.08(m,5H),0.80(t,3H)ppm; LCMS: 1000.5[M+1] + .

[0240] Step 9: (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((3-fluoropyridin-4-yl)methyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (compound 4-2)

[0241] The synthesis of compound 4-2 followed the steps in Example 1, using compound int-3b (2.5 g) as the starting material, and replacing the third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid with compound 4i. Compound 4-2 was a white solid (176 mg, yield: 5.8%). 1 H NMR(500MHz,DMSO-d6)δ10.80(s,1H),8.61(d,1H),8.45(d,2H),8.29(d,1H),8.24 (t,2H),8.15(s,1H),8.03(d,1H),7.98(t,2H),7.86(d,1H),7.60–7.55(m,2H),7. 52(t,1H),7.33(d,1H),7.26-7.21(m,1H),7.13(d,1H),7.07(d,1H),7.04(d,1H), 6.98(t,1H),4.64–4.54(m,1H),4.53–4.48(m,1H),4.44(dd,1H),4.30–4.14(m,3H) ,3.25–3.22(m,1H),3.16–3.08(m,3H),3.08–2.93(m,6H),2.90–2.82(m,2H),2.82 –2.75(m,1H),2.75–2.67(m,1H),2.04(t,J=7.7Hz,2H),1.96–1.88(m,1H),1.84(s ,3H),1.80–1.72(m,1H),1.69–1.61(m,1H),1.61–1.53(m,1H),1.50–1.40(m,2H), 1.39–1.27(m,6H),1.27–1.15(m,5H),0.83(t,J=6.7Hz,3H)ppm; LCMS: 1000.5[M+1] + .

[0242] Example 5

[0243] (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-([1,2,4]triazolo[1,5-a]pyridin-7-ylmethyl)-8-(3-guanidinopropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic dihexasaccharide-22-yl)-2-acetaminohexanamide (Compound 5)

[0244] Step 1: (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-([1,2,4]triazolo[1,5-a]pyridin-7-yl)methyl propionate (5b)

[0245] Under nitrogen protection, zinc powder (1.3 g, 19.94 mmol), iodine (254 mg, 0.998 mmol), and anhydrous DMF (10 mL) were added sequentially to a 100 mL reaction tube. The reaction was allowed to proceed at room temperature until the solution changed from brown to colorless. Then, a DMF solution of compound 5a (3.0 g, 6.65 mmol) and iodine (254 mg, 0.998 mmol) (30 mL) was added, and the reaction was allowed to proceed at room temperature until the solution changed from brown to colorless, and the reaction was allowed to continue for 1.5 h. After 1.5 h, 7-bromo-[1,2,4]thiazo[1,5-A]pyridine (1.58 g, 7.98 mmol), tris(dibenzylacetone)palladium (183 mg, 0.20 mmol), and 2-dicyclohexylphosphine-2′,6′-dimethoxy-biphenyl (274 mg, 0.665 mmol) were added, and the temperature was raised to 60 °C and the reaction was allowed to proceed for 6 h. The reaction was monitored by TLC until it ended. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 5b, a white solid (1.86 g, yield: 63.5%). LCMS: 443.2 [M+1] + .

[0246] Step 2: (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-([1,2,4]triazolo[1,5-a]pyridin-7-yl)propionic acid (5c)

[0247] In a 100 mL round-bottom flask, compound 5b (1.8 g, 4.07 mmol), trimethyltin hydroxide (2.2 g, 12.22 mmol), and 1,2-dichloroethane (20 mL) were added sequentially, and the mixture was refluxed for 6 h. The reaction was monitored by TLC until completion. After cooling to room temperature, the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 5c, a white solid (1.69 g, yield: 97.1%). LCMS: 429.1 [M+1] + .

[0248] Step 3: (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-([1,2,4]triazolo[1,5-a]pyridin-7-ylmethyl)-8-(3-guanidinopropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic dihexasaccharide-22-yl)-2-acetaminohexanamide (compound 5)

[0249] Compound 5 was synthesized following the steps in Example 1, using compound int-3b (2.5 g) as the starting material, and replacing the third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid with compound 5c. Compound 5 was a white solid (110 mg, yield: 3.6%). 1H NMR(400MHz,DMSO-d6)δ14.15(s,2H),10.82(s,1H),8.92(d,2H),8.57(s,1H),8 .50(d,2H),8.28(d,1H),8.03(d,1H),7.95(s,1H),7.91(d,1H),7.73(s,1H),7. 67–7.58(m,2H),7.54(d,2H),7.29(d,1H),7.23(d,2H),7.10(s,1H),7.05(t,1H ),6.99(t,1H),4.77(t,1H),4.54–4.45(m,1H),4.43–4.34(m,1H),4.25–4.13(m, 2H),4.07–3.97(m,1H),3.69–3.54(m,4H),3.41–3.26(m,3H),3.25–3.13(m,2H) ,3.09(d,2H),3.05–2.98(m,1H),2.93(d,3H),2.83–2.73(m,1H),2.71–2.59(m,1 H),2.30–2.19(m,1H),2.17–2.07(m,1H),1.91(s,1H),1.86(s,3H),1.73–1.55( m,2H),1.54–1.31(m,8H),1.30–1.07(m,5H),0.82(t,3H)ppm; LCMS: 1022.5[M+1] + .

[0250] Example 6

[0251] (2S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-8-(3-guanidinopropyl)-14-methyl-5-(naphth-2-ylmethyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 6)

[0252] Compound 6 was synthesized using compound int-1h (1.4 g) as the starting material, following the synthetic procedure of Example 1. The third amino acid, (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid, was replaced with compound Fmoc-3-(2-naphthyl)-D-alanine. Compound 6 was obtained as a white solid (48 mg, yield: 2.7%). 1H NMR(400MHz,DMSO-d6)δ8.57(d,1H),8.44(d,2H),8.36–8.22(m,1H),8.22–8.15(m,1H),8.14–8.07(m,1H),8.01(d,1H),7.96–7.78(m,4H ),7.71(d,1H),7.61(d,2H),7.47(d,6H),7.37(dd,1H),7.16(d,1H),7.07(d,1H),7.03–6.96(m,1H),6.64(d,1H),4.58–4.35(m,3H),4.32 –4.14(m,2H),3.70(d,2H),3.54–3.45(m,6H),3.33–3.11(m,6H),3.05(d,2H),3.00–2.82(m,3H),2.71(d,1H),2.12–1.91(m,2H),1.89–1 .80(m,3H),1.61(s,2H),1.51–1.37(m,2H),1.37–1.28(m,3H),1.25(d,5H),1.04(d,1H),0.91(d,1H),0.84(s,3H)ppm; LCMS: 1045.6[M+1] + .

[0253] Example 7

[0254] (2S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-5-(pyridin-4-ylmethyl)-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 7)

[0255] Compound 7 was synthesized using compound int-1h (2.4 g) as the starting material, following the synthetic procedure of Example 1. The third amino acid, (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid, was replaced with compound Fmoc-3-(4-pyridyl)-D-alanine. Compound 7 was obtained as a white solid (26 mg, yield: 0.9%). 1H NMR(500MHz,DMSO-d6)δ10.78(s,1H),8.53(d,1H),8.29(s,1H),8.16(d,1H),8.1 1(d,1H),8.08-8.01(m,2H),7.98(d,1H),7.59(d,1H),7.52(s,1H),7.47(s,1H), 7.39(d,1H),7.33(d,1H),7.28(dd,3H),7.14(s,1H),7.06(t,1H),6.99(s,1H),6 .97(d,1H),6.62(s,1H),4.45(d,1H),4.37(dd,2H),4.30–4.24(m,1H),4.24–4.15 (m,2H),3.70–3.60(m,2H),3.16–3.09(m,2H),3.07–2.96(m,6H),2.95–2.86(m,2 H),2.83–2.73(m,1H),2.73–2.66(m,1H),2.64(d,1H),2.40–2.34(m,1H),2.06–1 .95(m,2H),1.89(s,1H),1.83(s,3H),1.80–1.67(m,2H),1.65–1.55(m,1H),1.42 (d,3H),1.27(s,4H),1.23(d,4H),1.02(d,3H),0.82(t,3H)ppm; LCMS: 996.5[M+1] + .

[0256] Example 8

[0257] (2S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(3,4-difluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 8)

[0258] Compound 8 was synthesized using compound int-1h (2.4 g) as the starting material, following the synthetic procedure of Example 1. The third amino acid, (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid, was replaced with compound FMOC-D-3,4-difluorophenylalanine. Compound 8 was obtained as a white solid (33 mg, yield: 1.1%). 1H NMR(500MHz,DMSO-d6)δ10.78(s,1H),8.53(d,1H),8.29(s,1H),8.16(d,1H),8.1 1(d,1H),8.08–8.01(m,2H),7.98(d,1H),7.59(d,1H),7.52(s,1H),7.47(s,1H), 7.39(d,1H),7.33(d,1H),7.28(dd,3H),7.14(s,1H),7.06(t,1H),6.99(s,1H),6 .97(d,1H),6.62(s,1H),4.45(d,1H),4.37(dd,2H),4.30–4.24(m,1H),4.24–4.15 (m,2H),3.70–3.60(m,2H),3.16–3.09(m,2H),3.07–2.96(m,6H),2.95–2.86(m,2 H),2.83–2.73(m,1H),2.73–2.66(m,1H),2.64(d,H),2.40–2.34(m,1H),2.06–1. 95(m,2H),1.89(s,1H),1.83(s,3H),1.80–1.67(m,2H),1.65–1.55(m,1H),1.42( d,3H),1.27(s,4H),1.23(d,4H),1.02(d,3H),0.82(t,3H)ppm; LCMS: 1031.5[M+1] + .

[0259] Example 9

[0260] (2S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclotrialkyl-22-yl)-2-acetamido-5-(1H-imidazol-1-yl)pentanamide (Compound 9)

[0261] Step 1: (S)-2-((tert-Butoxycarbonyl)amino)-5-(1H-imidazol-1-yl)tert-butyl valerate (9b)

[0262] In a 100 mL round-bottom flask, compound 9a (5.0 g, 14.19 mmol), potassium carbonate (3.92 g, 28.39 mmol), and imidazole (1.16 g, 17.03 mmol) were dissolved in DMF (50 mL), and the mixture was heated to 60 °C and reacted for 6 h. The reaction was monitored by LCMS until completion. After cooling to room temperature, the organic solvent was removed under reduced pressure. The residue was purified by column chromatography (PE:EA = 9:1) to give compound 9b, a colorless oily solid (2.7 g, yield: 56%). LCMS: 340.2 [M+1] + .

[0263] Step 2: (S)-2-((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-(1H-imidazol-1-yl)valeric acid (9c)

[0264] In a 100 mL round-bottom flask, compound 9b (2.7 g, 7.96 mmol) was dissolved in dichloromethane (30 mL), and dioxane hydrochloride solution (10 mL, 4 M) was added dropwise under ice bath. The reaction was carried out at room temperature for 1 h. After TLC monitoring showed the reaction was complete, the solvent was removed under reduced pressure. The residue was dissolved in a mixture of tetrahydrofuran and water (15 / 15 mL), and sodium carbonate (2.53 g, 23.89 mmol) and 9-fluorenylmethyl-N-succinimide carbonate (3.22 g, 9.55 mmol) were added under ice bath. The reaction was carried out at room temperature for 3 h. After TLC monitoring showed the reaction was complete, the mixture was extracted with ethyl acetate, dried over anhydrous sulfuric acid, filtered, and the organic solvent was removed under reduced pressure. The residue was purified by column chromatography (PE:EA = 1:1) to give compound 9c, a white solid (2.1 g, yield: 65.2%). LCMS: 405.2 [M+1] + .

[0265] Step 3: (2S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclotrialkyl-22-yl)-2-acetamido-5-(1H-imidazol-1-yl)pentanamide (Compound 9)

[0266] Compound 9 was synthesized using compound int-1h (2.4 g) as the starting material, following the synthetic procedure in Example 1. The sixth amino acid, fluorenemethoxycarbonyl acyl leucine, was replaced with compound 9c. Compound 9 was obtained as a white solid (80 mg, yield: 2.6%). 1H NMR(400MHz,DMSO-d6)δ10.82(d,1H),9.10(d,1H),8.96(s,1H),8.66–8.40(m,1H) ),8.34(d,1H),8.30–8.19(m,1H),8.12(s,1H),8.10–8.03(m,1H),7.91(s,1H),7. 80(d,1H),7.72(d,2H),7.60(dd,2H),7.34(dd,1H),7.29–7.19(m,3H),7.18–7.1 2(m,2H),7.08(t,3H),7.01(d,1H),4.72–4.53(m,2H),4.52–4.38(m,1H),4.37–4. 28(m,1H),4.20(s,3H),3.88–3.72(m,4H),3.53–3.25(m,6H),3.21–3.09(m,1H), 3.02(s,4H),2.92(d,1H),2.83–2.63(m,2H),2.26–2.13(m,1H),2.01(d,1H),1.88 (t,3H),1.79(d,2H),1.64(d,2H),1.52–1.41(m,2H),1.32(d,J=11.2Hz,5H),1.20 –1.08(m,1H),1.04(d,1H),0.99(d,1H),0.89–0.80(m,1H)ppm; LCMS: 1065.5[M+1] + .

[0267] Example 10

[0268] (S)-N-((2S,5R,8S,11S,22S)-2-((1H-1,2,3-triazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 10)

[0269] Step 1: (S)-2-((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(1-benzyl-1H-1,2,3-triazol-4-yl)propionic acid (10b)

[0270] In a 250 mL round-bottom flask, compound 10a (5.0 g, 14.91 mmol), benzyl azide (2.1 g, 15.65 mmol), anhydrous copper sulfate (119 mg, 0.746 mmol), sodium vitamin C (5.9 g, 29.82 mmol), and DMF (80 mL) were added sequentially, and the mixture was reacted overnight at room temperature. After the reaction was monitored by TLC, the mixture was quenched with water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The residue was purified by column chromatography (PE:EA = 1:1) to give compound 10b, a white solid (5.8 g, yield: 83.1%). LCMS: 469.2 [M+1] + .

[0271] Step 2: (S)-N-((2S,5R,8S,11S,22S)-11-((1H-indol-3-yl)methyl)-2-((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (10c)

[0272] Compound 10c was synthesized using intermediate 3 (2.4 g) as the starting material, following the synthetic procedure in Example 1. The fourth amino acid, N-Fmoc-N'-triphenylmethyl-L-histidine, was replaced with compound 10b. Compound 10c was obtained as a white solid (398 mg, yield: 12.7%). LCMS: 1090.6 [M+1] + .

[0273] Step 3: (S)-N-((2S,5R,8S,11S,22S)-2-((1H-1,2,3-triazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinylpropyl)-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 10)

[0274] Under a hydrogen atmosphere, compound 10c (398 mg, 0.365 mmol), palladium on carbon (200 mg, 10% Pd content, 50% w / w), and methanol (10 mL) were added sequentially to a 100 mL round-bottom flask, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by LCMS until completion. The organic solvent was removed by filtration and under reduced pressure. The residue was passed through a refining apparatus to obtain compound 10, a white solid (152 mg, yield: 41.6%). 1H NMR(500MHz,DMSO-d6)δ10.79(s,1H),8.60(d,1H),8.49(d,1H),8.23(d,1H),7.99(dd,2H),7.79(s,1H),7.55(d,2H),7.38(s,1H),7.31 (d,2H),7.20(dd,2H),7.05(dt,3H),6.95(t,1H),4.53(d,1H),4.41(d,1H),4.31(dd,2H),4.24–4.18(m,1H),4.16–4.10(m,1H),3.27–3. 19(m,2H),3.17(d,1H),3.14(d,1H),3.03(s,2H),3.01(d,2H),2.97(d,2H),2.92(dd,2H),2.84(d,1H),2.78(dd,2H),2.09–1.99(m,2H), 1.85(s,3H),1.83(s,3H),1.78(dd,2H),1.66–1.53(m,2H),1.46(dd,1H),1.34(dd,5H),1.22(t,5H),0.82(t,3H)ppm; LCMS: 1000.5[M+1] + .

[0275] Example 11

[0276] (S)-N-((2S,5R,8S,11S,14R,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(2,4-difluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 11)

[0277] Step 1: (R)-2-((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2,4-difluorophenyl)propionate methyl ester (11a)

[0278] Under nitrogen protection, zinc powder (1.3 g, 19.94 mmol), iodine (254 mg, 0.998 mmol), and anhydrous DMF (10 mL) were added sequentially to a 100 mL reaction tube. The reaction was allowed to proceed at room temperature until the solution changed from brown to colorless. Then, a DMF solution of compound 5a (3.0 g, 6.65 mmol) and iodine (254 mg, 0.998 mmol) (30 mL) was added, and the reaction was allowed to proceed at room temperature until the solution changed from brown to colorless, and the reaction was allowed to continue for 1.5 h. After 1.5 h, 1-bromo-2,4-difluorobenzene (1.92 g, 9.95 mmol), tris(dibenzylacetone)palladium (183 mg, 0.20 mmol), and 2-dicyclohexylphosphine-2′,6′-dimethoxy-biphenyl (274 mg, 0.665 mmol) were added, and the temperature was raised to 60 °C and the reaction was allowed to proceed for 6 h. The reaction was monitored by TLC until it ended. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 9:1) to give compound 11a, a white solid (1.76 g, yield: 61.5%). LCMS: 438.2 [M+1] + .

[0279] Step 2: (R)-2-((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2,4-difluorophenyl)propionic acid (11b)

[0280] In a 100 mL round-bottom flask, compound 11a (1.75 g, 4.00 mmol), trimethyltin hydroxide (2.16 g, 12.01 mmol), and 1,2-dichloroethane (20 mL) were added sequentially, and the mixture was refluxed for 6 h. After the reaction was complete as monitored by TLC, the mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 11b, a white solid (1.69 g, yield: 100%). LCMS: 424.1 [M+1] + .

[0281] Step 3: (S)-N-((2S,5R,8S,11S,14R,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(2,4-difluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 11)

[0282] Compound 11 was synthesized according to the steps in Example 1, starting with intermediate 2b (1.2 g) as the starting amino acid, and replacing the third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid with compound 11b. Compound 11 was a white solid (68 mg, yield: 4.6%). 1 H NMR(400MHz,DMSO-d6)δ14.11(s,2H),10.82(s,1H),8.95(s,1H),8.51(s,1H),8.39(d, 1H),8.06(d,1H),7.96(t,2H),7.75(d,2H),7.56–7.47(m,2H),7.40–7.31(m,1H),7.26( d,1H),7.24(s,1H),7.19(d,1H),7.08(s,1H),7.04(t,2H),6.98(t,2H),6.92(d,1H),4 .81–4.65(m,1H),4.39(t,1H),4.33–4.26(m,1H),4.19–4.08(m,2H),4.03–3.91(m,1H), 3.69–3.57(m,1H),3.30–3.24(m,2H),3.24–3.15(m,2H),3.13–3.06(m,2H),3.06–3.01 (m,2H),2.98–2.88(m,1H),2.79(t,2H),2.71–2.60(m,1H),2.24–2.09(m,1H),2.04(dd, 1H),1.87(s,3H),1.66–1.53(m,3H),1.52–1.46(m,1H),1.40(s,1H),1.37(s,4H),1.25( s,3H),1.24–1.15(m,3H),1.03(d,3H),0.87(d,1H),0.82(t,3H)ppm; LCMS: 1031.5[M+1] + .

[0283] Example 12

[0284] (S)-N-((2S,5R,8S,11S,14R,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(2-chloro-4-fluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 12)

[0285] Step 1: (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2-chloro-4-fluorophenyl)propionate (12a)

[0286] Under nitrogen protection, zinc powder (1.3 g, 19.94 mmol), iodine (254 mg, 0.998 mmol), and anhydrous DMF (10 mL) were added sequentially to a 100 mL reaction tube. The reaction was allowed to proceed at room temperature until the solution changed from brown to colorless. Then, a DMF solution of compound 5a (3.0 g, 6.65 mmol) and iodine (254 mg, 0.998 mmol) (30 mL) was added, and the reaction was allowed to proceed at room temperature until the solution changed from brown to colorless, and the reaction was allowed to continue for 1.5 h. After 1.5 h, 1-bromo-2-chloro-4-fluorobenzene (2.05 g, 9.78 mmol), tris(dibenzylacetone)palladium (183 mg, 0.20 mmol), and 2-dicyclohexylphosphine-2′,6′-dimethoxy-biphenyl (274 mg, 0.665 mmol) were added, and the temperature was raised to 60 °C and the reaction was allowed to proceed for 6 h. The reaction was monitored by TLC until it ended. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 9:1) to give compound 12a, a white solid (1.79 g, yield: 59.5%). LCMS: 454.1 [M+1] + .

[0287] Step 2: (R)-2-((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(2-chloro-4-fluorophenyl)propionic acid (12b)

[0288] In a 100 mL round-bottom flask, compound 12a (1.78 g, 3.93 mmol), trimethyltin hydroxide (2.12 g, 11.79 mmol), and 1,2-dichloroethane (20 mL) were added sequentially, and the mixture was refluxed for 6 h. After the reaction was complete as monitored by TLC, the mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 12b, a white solid (1.67 g, yield: 96.5%). LCMS: 439.1 [M+1] + .

[0289] Step 3: (S)-N-((2S,5R,8S,11S,14R,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(2-chloro-4-fluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 12)

[0290] Compound 12 was synthesized according to the steps in Example 1, starting with intermediate 2b (1.2 g) as the starting amino acid, and replacing the third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid with compound 12b. Compound 12 was obtained as a white solid (48 mg, yield: 3.2%). 1 H NMR(500MHz,DMSO-d6)δ14.11(s,1H),10.80(s,1H),8.91(s,1H),8.39–8.20(m,1H),8 .06(d,1H),7.99(d,1H),7.95(d,1H),7.81(d,1H),7.72(s,1H),7.55–7.47(m,2H),7. 41(dd,1H),7.38–7.34(m,1H),7.27(d,1H),7.22(s,1H),7.16(t,1H),7.09(s,1H),7. 06–7.00(m,1H),6.96(dd,2H),4.69(s,1H),4.37–4.25(m,2H),4.20–4.08(m,2H),4.06 –3.96(m,1H),3.69–3.56(m,1H),3.34–3.30(m,4H),3.29–3.21(m,2H),3.17(t,2H),3 .12–2.98(m,3H),2.95–2.89(m,1H),2.87–2.80(m,1H),2.75–2.63(m,1H),2.17–2.02 (m,2H),1.87(s,3H),1.85–1.80(m,1H),1.72–1.60(m,1H),1.59–1.49(m,3H),1.43(d ,2H),1.40–1.30(m,4H),1.23(d,5H),1.02(d,3H),0.82(t,3H)ppm; LCMS: 1047.5[M+1] + .

[0291] Example 13

[0292] (S)-N-((2S,5R,8S,11S,14R,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclotrialkyl-22-yl)-2-acetaminohexanamide (Compound 13)

[0293] Compound 13 was synthesized according to the steps in Example 1, using intermediate 2a (1.2 g) to obtain compound 13 as a white solid (26 mg, yield: 1.8%). 1 H NMR(400MHz,DMSO-d6)δ10.85(s,1H),8.53(s,1H),8.41(s,2H),8.26(s,1H),8.19(s,2H),8.12(d,1H),8.04(d,1H),7.60 (d,2H),7.50(s,3H),7.45(d,1H),7.35(d,1H),7.27–7.19(m,2H),7.16(s,1H),7.07(t,3H),7.00(t,1H),6.64(s,1H),4. 46(d,1H),4.38(d,1H),4.30(dd,1H),4.21(s,2H),3.22–3.10(m,5H),3.03(d,8H),2.85–2.74(m,2H),2.70(s,2H),2.01( s,2H),1.85(s,3H),1.74(s,1H),1.61(s,1H),1.47(s,2H),1.28(d,9H),1.03(d,3H),0.84(s,3H)ppm; LCMS: 1013.5[M+1] + .

[0294] Example 14

[0295] (S)-N-((2S,5R,8S,11S,14S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexano-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 14)

[0296] Compound 14 was synthesized according to the steps in Example 1, using intermediate 2b (1.2 g) to obtain compound 14 as a white solid (31 mg, yield: 2.1%). 1H NMR(400MHz,DMSO-d6)δ10.85(s,1H),8.53(s,1H),8.41(s,2H),8.26(s,1H),8.19(s,2H),8.12(d,1H),8.04(d,1H),7.60(d,2H),7 .50(s,3H),7.45(d,1H),7.35(d,1H),7.27–7.19(m,2H),7.16(s,1H),7.07(t,3H),7.00(t,1H),6.64(s,1H),4.46(d,1H),4.42–4.3 5(m,1H),4.34–4.25(m,1H),4.24–4.14(m,2H),3.22–3.10(m,5H),3.10–2.88(m,8H),2.85–2.74(m,2H),2.74–2.61(m,2H),2.09–1 .94(m,2H),1.85(s,3H),1.74(s,1H),1.68–1.56(m,1H),1.47(s,2H),1.28(d,9H),1.03(d,3H),0.84(s,3H)ppm; LCMS: 1013.5[M+1] + .

[0297] Example 15

[0298] Step 1: 5-Amino-5-methylhexanoic acid (15b)

[0299] In a 100 mL round-bottom flask, compound 15a (2 g, 15.72 mmol) was dissolved in 20 mL of 6 M HCl, and the mixture was refluxed for 24 h. The reaction was monitored by LC-MS until completion. After cooling to room temperature, the solvent was removed under reduced pressure to obtain compound 15b, a white solid (2.1 g, yield: 92.1%). The crude product was used directly in the next step without purification. LC-MS: 146.1 [M+1] + .

[0300] Step 2: 5-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-methylhexanoic acid (15c)

[0301] In a 100 mL round-bottom flask, compound 15b (2.1 g, 14.48 mmol) and sodium carbonate (4.6 g, 43.45 mmol) were dissolved in tetrahydrofuran / water (20 mL / 20 mL). A tetrahydrofuran solution of 9-fluorenylmethyl-N-succinimide carbonate (5.86 g, 17.38 mmol) was added dropwise under ice bath conditions. After the addition was complete, the reaction was allowed to proceed at room temperature for 3 h. The reaction was monitored by TLC until completion. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The residue was purified by column chromatography (PE:EA = 5:1) to give compound 15c as a white solid (4.08 g, yield: 76.8%). LCMS: 368.1 [M+1] + .

[0302] Step 3: (9H-fluorene-9-yl)methyl(5-amino-2-methylpentan-2-yl)carbamate (15d)

[0303] In a 500 mL round-bottom flask, compound 15c (4.0 g, 10.90 mmol), triethylamine (1.43 g, 14.17 mmol), and N,N-dimethylacetamide (40 mL) were added sequentially. Diphenyl azidophosphate (3.90 g, 14.17 mmol) was added dropwise under ice bath conditions. The reaction was carried out at room temperature for 3 h. TLC monitoring showed the starting material had disappeared. 40 mL of 3 M hydrochloric acid solution was added, and the temperature was raised to 60 °C for another 6 h. LCMS monitoring indicated the reaction was complete. The mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 15d, a colorless oily solid (2.38 g, yield: 64.6%). LCMS: 339.2 [M+1] + .

[0304] Step 4:

[0305] Compound 15e was synthesized following the method used for compound int-1h. Using 2-CTC resin as a starting material, compound 15e was obtained by coupling and deprotection with compound 15d, N-(9-fluorenylmethoxycarbonyl)-N1-tert-butoxycarbonyl-L-tryptophan, and N-fluorenylmethoxycarbonyl-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl-L-arginine, respectively. The resulting pale yellow solid (9.98 g) was LCMS: 811.4 [M+1]. + .

[0306] Step 5: (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinopropyl)-14,14-dimethyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 15)

[0307] The synthesis of compound 15 was carried out in accordance with Example 1. Starting from compound 15e (1.2 g), compound 15 was obtained by coupling, deprotection, acetylation, cleavage and cyclization, respectively, resulting in a white solid (49 mg, yield: 3.2%). 1 H NMR(500MHz,DMSO-d6)δ10.80(s,1H),8.93(s,1H),8.52(d,1H),8.32(d,1H),8.1 3(d,1H),8.02(d,1H),7.96(d,1H),7.86(d,1H),7.60(d,1H),7.50(dd,J=12.5,6 .7Hz,2H),7.35–7.27(m,2H),7.24–7.19(m,2H),7.15(d,2H),7.10–7.03(m,3H), 6.97(t,1H),4.69–4.56(m,1H),4.51–4.42(m,2H),4.25–4.18(m,1H),4.14(t,2H) ,3.49–3.45(m,5H),3.40–3.35(m,3H),3.16–3.06(m,2H),3.04–2.94(m,4H),2.9 3–2.84(m,1H),2.83–2.73(m,2H),2.05(dd,2H),1.93–1.86(m,1H),1.84(s,3H), 1.82–1.74(m,1H),1.72–1.63(m,1H),1.62–1.52(m,1H),1.48–1.33(m,3H),1.32 –1.19(m,8H),1.16(s,3H),1.05(d,3H),0.87–0.79(m,3H)ppm; LCMS: 1027.6[M+1] + .

[0308] Example 16

[0309] (S)-N-((2S,5R,8S,11S,14S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(2,4-difluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 16)

[0310] Compound 16 was synthesized following the steps in Example 1, using intermediate 2b (1.2 g) to obtain a white solid (71 mg, yield: 5.3%). 1 H NMR(500MHz,DMSO-d6)δ14.06(s,1H),10.79(s,1H),8.92(s,1H),8.38(d,1H),8.31(s,1H),7.98( d,1H),7.91(d,1H),7.84(d,1H),7.76–7.69(m,2H),7.53(d,1H),7.49(s,1H),7.43–7.35(m,1H), 7.31(d,J=8.1Hz,1H),7.25(s,1H),7.22–7.15(m,1H),7.09(s,1H),7.06-7.01(m,2H),6.96(t,1H ),4.59(s,1H),4.51–4.42(m,1H),4.37–4.29(m,1H),4.24–4.14(m,2H),4.11–4.04(m,1H),3.79–3 .69(m,1H),3.48–3.41(m,4H),3.33–3.29(m,3H),3.28–3.21(m,2H),3.20–3.12(m,2H),3.11–3.0 1(m,3H),2.96–2.88(m,2H),2.86–2.78(m,2H),2.70–2.59(m,1H),2.37(s,1H),2.21–2.12(m,1H) ,2.12–1.95(m,1H),1.90–1.81(m,4H),1.74-1.65(m,1H),1.63-1.55(m,1H),1.46(d,3H),1.39–1 .29(m,3H),1.24(s,2H),1.21–1.12(m,3H),0.87–0.82(m,3H),0.80(d,2H)ppm; LCMS: 1031.5[M+1] + .

[0311] Example 17

[0312] (S)-N-((2S,5R,8S,11S,14S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(2-chloro-4-fluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-acetaminohexanamide (Compound 17)

[0313] Compound 17 was synthesized following the steps in Example 1, using intermediate 2a (1.2 g) and compound 12b as starting materials to obtain compound 17, a white solid (60 mg, yield: 4.0%). 1 H NMR(500MHz,DMSO-d6)δ14.05(s,1H),10.79(s,1H),8.91(s,1H),8.34(d,1H),8.14(s,1H),8.0 2-7.92(m,1H),7.87(d,1H),7.71(d,1H),7.59(s,1H),7.54(d,1H),7.42(dd,2H),7.31(d,1H),7 .24(s,1H),7.21–7.14(m,1H),7.10(d,1H),7.05(t,H),6.96(t,1H),4.58(s,1H),4.49–4.42(m, 1H),4.29(t,1H),4.25–4.18(m,1H),4.17–4.08(m,1H),3.81–3.68(m,1H),3.49–3.40(m,2H),3. 30–3.20(m,4H),3.19–3.11(m,2H),3.10–3.02(m,3H),3.01–2.88(m,3H),2.88–2.80(m,1H),2.7 7–2.62(m,1H),2.48–2.43(m,1H),2.37(s,1H),2.21–2.12(m,1H),2.12–2.04(m,1H),2.03–1.95 (m,1H),1.90(d,1H),1.84(s,3H),1.74–1.64(m,1H),1.64–1.56(m,1H),1.56–1.39(m,4H),1.39 –1.28(m,3H),1.24(s,4H),1.20(dd,3H),0.88–0.82(m,3H),0.80(d,2H)ppm; LCMS: 1047.5[M+1] + .

[0314] Example 18

[0315] (S)-N-((2S,5R,8S,11S,14S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(2-chloro-4-fluorobenzyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-(2,2,2-trifluoroacetamido)hexanoamide (Compound 18)

[0316] Compound 18 was synthesized following the steps in Example 1, using intermediate 2b (600 mg) and compound 12b as starting materials, and replacing the acetyl group with a trifluoroacetyl group to obtain compound 18, a white solid (20 mg, yield: 2.7%). 1 H NMR(500MHz,DMSO-d6)δ10.78(s,1H),8.53(d,1H),8.29(s,1H),8.16(d,1H),8.1 1(d,1H),8.08-8.01(m,2H),7.98(d,1H),7.59(d,1H),7.52(s,1H),7.47(s,1H), 7.39(d,1H),7.33(d,1H),7.28(dd,3H),7.14(s,1H),7.06(t,1H),6.99(s,1H),6 .97(d,1H),6.62(s,1H),4.45(d,1H),4.37(dd,2H),4.30–4.24(m,1H),4.24–4.15 (m,2H),3.70–3.60(m,2H),3.16–3.09(m,2H),3.07–2.96(m,6H),2.95–2.86(m,2 H),2.83–2.73(m,1H),2.73–2.66(m,1H),2.64(d,1H),2.40–2.34(m,1H),2.06–1. 95(m,2H),1.89(s,1H),1.83(s,3H),1.80–1.67(m,2H),1.65–1.55(m,1H),1.42( d,3H),1.27(s,4H),1.23(d,4H),1.02(d,3H),0.82(t,3H)ppm; LCMS: 1011.5[M+1] + .

[0317] Example 19

[0318] (S)-N-((2S,5R,8S,11S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-(4-fluorobenzyl)-8-(3-guanidinopropyl)-14,14-dimethyl-3,6,9,12,19,23-hexaoxo-1,4,7,10,13,18-hexaazacyclic trisaccharide-22-yl)-2-(2,2,2-trifluoroacetamido)hexanoamide (compound 19)

[0319] Compound 19 was synthesized following the steps in Example 1, using compound 15f (600 mg) as the starting material and replacing the acetyl group with a trifluoroacetyl group to obtain compound 19, a white solid (15 mg, yield: 2.5%). 1 H NMR(500MHz,DMSO-d6)δ14.03(s,2H),10.67–10.44(m,1H),8.93(s,1H),8.52(s ,1H),8.31(t,1H),8.17–7.99(m,2H),7.99–7.83(m,2H),7.51(d,1H),7.47(s,2 H),7.27(t,1H),7.23–7.18(m,2H),7.13(s,1H),7.09–7.04(m,2H),7.01–6.90( m,1H),4.71–4.52(m,1H),4.50–4.41(m,1H),4.18(d,2H),4.14(d,1H),3.34–3.2 7(m,2H),3.14–2.94(m,6H),2.92–2.72(m,3H),2.04(d,1H),2.02–1.96(m,1H), 1.94–1.87(m,1H),1.84(s,3H),1.82–1.75(m,1H),1.74–1.67(m,1H),1.65(s,3H ),1.61–1.53(m,1H),1.43(s,4H),1.32(s,4H),1.27–1.20(m,7H),1.17(d,1H), 1.13(s,2H),1.05(d,1H),1.02(s,1H),0.87–0.78(m,3H)ppm; LCMS: 1081.5[M+1] + .

[0320] Example 20

[0321] Step 1: 4-(1-Aminocyclopropyl)butyric acid (20b)

[0322] In a 100 mL round-bottom flask, compound 20a (5 g, 39.95 mmol) and potassium hydroxide (13.5 g, 239.67 mmol) were dissolved in a mixture of ethanol and water (25 / 25 mL), and the mixture was refluxed for 24 h. The reaction was monitored by LCMS until completion. After cooling to room temperature, the solvent was removed under reduced pressure to obtain compound 20b. The crude product was used directly in the next step without purification. LCMS: 144.1 [M+1] + Step 2: 4-(1-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)cyclopropyl)butyric acid (20c)

[0323] In a 250 mL round-bottom flask, compound 20b (5.7 g, 39.95 mmol) and sodium bicarbonate (6.7 g, 79.9 mmol) were dissolved in tetrahydrofuran / water (50 mL / 50 mL). A tetrahydrofuran solution of 9-fluorenylmethyl-N-succinimide carbonate (13.5 g, 39.95 mmol) was added dropwise under ice bath conditions. After the addition was complete, the reaction was allowed to proceed at room temperature for 3 h. The reaction was monitored by TLC until completion. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, and the organic solvent was removed under reduced pressure. The residue was purified by column chromatography (PE:EA = 5:1) to give compound 20c as a white solid (4.7 g, yield: 32.2%). LCMS: 366.2 [M+1] + .

[0324] Step 3: (9H-fluorene-9-yl)methyl (1-(3-aminopropyl)cyclopropyl)carbamate (20d)

[0325] In a 250 mL round-bottom flask, compound 20c (4.7 g, 12.88 mmol), triethylamine (1.69 g, 16.74 mmol), and N,N-dimethylacetamide (50 mL) were added sequentially. Diphenyl azidophosphate (4.6 g, 16.74 mmol) was added dropwise under ice bath conditions. The reaction was carried out at room temperature for 3 h. TLC monitoring showed the starting material had disappeared. 50 mL of 3 M hydrochloric acid solution was added, and the temperature was raised to 60 °C for another 6 h. LCMS monitoring indicated the reaction was complete. The mixture was cooled to room temperature, and the organic solvent was removed under reduced pressure. The residue was subjected to column chromatography (PE:EA = 1:1) to give compound 20d, a colorless oily solid (4.06 g, yield: 93.5%). LCMS: 337.2 [M+1] + .

[0326] Step 4:

[0327] The synthesis of 20e followed that of compound int-1h. Using 2-CTC resin as a starting material, compound 20e was obtained by coupling and deprotection with compound 20d, N-(9-fluorenylmethoxycarbonyl)-N1-tert-butoxycarbonyl-L-tryptophan, and N-fluorenylmethoxycarbonyl-2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl-L-arginine, respectively. The resulting pale yellow solid (17.4 g) was LCMS: 809.4 [M+1]. + .

[0328] Step 5: (S)-N-((6S,9S,12R,15S,18S)-15-((1H-imidazol-4-yl)methyl)-6-((1H-indol-3-yl)methyl)-12-(4-fluorobenzyl)-9-(3-guanidinylpropyl)-5,8,11,14,17,21-hexano-4,7,10,13,16,22-hexaazacarbon[2.22]pentadosine-18-yl)-2-acetaminohexanamide (compound 20)

[0329] Compound 20 was synthesized according to Example 1. Starting from compound 20e (2.4 g), compound 20 was obtained by coupling, deprotection, acetylation, cleavage and cyclization, respectively, resulting in a white solid (66 mg, yield: 2.2%). 1 H NMR(500MHz,DMSO-d6)δ14.10(d,2H),10.80(s,1H),8.94(s,1H),8.49(d,1H),8.25(dd,2H),8.07(s,1H),7.98(d,2H),7.81(d,1H),7.64–7.4 6(m,3H),7.33(d,1H),7.06(dd,3H),6.97(t,2H),6.90(d,1H),6.62(d ,1H),4.64(dd,1H),4.47(dd,3H),4.42–4.34(m,1H),4.26–4.13(m,3H) ,3.14–3.00(m,5H),2.99–2.82(m,4H),2.76(dd,1H),2.65(dd,1H),2. 00(d,2H),1.94–1.79(m,4H),1.79–1.71(m,1H),1.70–1.61(m,2H),1.5 7(d,1H),1.50–1.41(m,2H),1.36(d,4H),1.23(d,6H),1.04(s,1H),0. 89–0.78(m,3H),0.45(d,1H),0.43–0.21(m,2H)ppm; LCMS: 1025.5[M+1] + .

[0330] Example 21

[0331] (S)-N-((6S,9S,12R,15S,18S)-15-((1H-imidazol-4-yl)methyl)-6-((1H-indol-3-yl)methyl)-9-(3-guanidinopropyl)-12-(naphthyl-2-ylmethyl)-5,8,11,14,17,21-hexaoxo-4,7,10,13,16,22-hexaazacarbon[2.22]pentadodecane-18-yl)-2-acetaminohexanamide (compound 21)

[0332] Compound 21 was synthesized according to the steps in Example 1. Starting from compound 20e (2.4 g), the compounds were coupled, deprotected, acetylated, cleaved, and cyclized to obtain compound 21, a white solid (122 mg, yield: 4.0%).

[0333] 1 H NMR(400MHz,DMSO-d6)δ14.23-13.83(m,2H),10.81(s,1H),8.84(s,1H),8. 60(d,1H),8.31–8.22(m,2H),8.06(s,1H),8.01–7.94(m,2H),7.87(d,2H),7 .82(d,2H),7.71(s,1H),7.61–7.52(m,2H),7.51–7.44(m,3H),7.40(d,1H), 7.35(d,1H),7.17–7.03(m,3H),7.02–6.95(m,2H),4.78–4.57(m,2H),4.45– 4.35(m,1H),4.30–4.15(m,3H),3.25–3.14(m,1H),3.11–2.83(m,7H),2.67( dd,1H),2.01(t,2H),1.93–1.81(m,4H),1.80–1.71(m,1H),1.67(d,1H),1.6 0–1.53(m,1H),1.52–1.30(m,6H),1.29–1.13(m,7H),1.12–1.00(m,1H),0.9 2–0.76(m,3H),0.52–0.44(m,1H),0.43–0.27(m,3H)ppm; LCMS: 1057.6[M+1] + .

[0334] Example 22

[0335] (S)-N-((2S,5R,8S,11S,14R,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-8-(3-guanidinopropyl)-14-methyl-5-(naphth-2-ylmethyl)-3,6,9,12,19,23-hexano-1,4,7,10,13,18-hexaazacyclic trimerose-22-yl)-2-acetaminohexanamide (compound 22)

[0336] Compound 22 was synthesized using compound int-2a (1.2 g) as the starting material, following the synthetic procedure of Example 1. The third amino acid, (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid, was replaced with Fmoc-3-(2-naphthyl)-D-alanine to give compound 22, a white solid (53 mg, yield: 3.5%). 1 H NMR(500MHz,DMSO-d6)δ14.01(d,1H),10.76(s,1H),8.83(s,1H),8.58(d,1H ),8.21(t,2H),8.04(d,1H),7.95(d,1H),7.85(d,1H),7.81(d,2H),7.77(d, 1H),7.71(s,1H),7.60(d,1H),7.52(s,1H),7.45(d,3H),7.39(d,2H),7.34( d,1H),7.15(s,1H),7.06(t,1H),6.99(dd,2H),4.61(s,2H),4.39(d,1H),4. 18(s,3H),3.64(s,1H),3.22–3.10(m,2H),3.08–2.96(m,3H),2.95–2.85(m, 4H),2.77–2.68(m,1H),2.01(d,1H),1.99–1.93(m,1H),1.87(s,1H),1.84(s ,3H),1.81–1.73(m,1H),1.64(s,1H),1.56(s,1H),1.45(s,1H),1.37(s,2H) ,1.29–1.12(m,12H),1.05–0.98(m,3H),0.82(s,3H)ppm; LCMS: 1045.6[M+1] + .

[0337] Example 23

[0338] (S)-N-((2S,5R,8S,11S,14S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-8-(3-guanidinopropyl)-14-methyl-5-(naphth-2-ylmethyl)-3,6,9,12,19,23-hexano-1,4,7,10,13,18-hexaazacyclic trimerose-22-yl)-2-acetaminohexanamide (compound 23)

[0339] Compound 23 was synthesized using compound int-2b (1.2 g) as the starting material, following the synthetic procedure of Example 1. The third amino acid, (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid, was replaced with Fmoc-3-(2-naphthyl)-D-alanine to give compound 23, a white solid (68 mg, yield: 4.5%). 1 H NMR(500MHz,DMSO-d6)δ13.95(d,2H),10.83(s,1H),8.84(s,1H),8.52(d,1H),8.28(d,1H),8.19(d,1H),7.96(s,3H),7.8 5(d,1H),7.82–7.74(m,3H),7.68(s,1H),7.58(d,1H),7.54–7.44(m,3H),7.41(s,1H),7.35(dd,2H),7.14(s,1H),7.06(s, 2H),6.98(t,1H),4.69(s,1H),4.57(s,1H),4.43(d,1H),4.20(dd,3H),3.69(s,1H),3.17(d,1H),3.11–2.81(m,8H),2.64( t,1H),1.98(s,3H),1.83(s,4H),1.76(s,1H),1.58(d,2H),1.43(s,2H),1.27(d,12H),0.83(t,6H)ppm; LCMS: 1045.6[M+1] + .

[0340] Example 24

[0341] (S)-N-((2S,5R,8S,11S,14R,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((2,3-dihydrobenzofuran-5-yl)methyl)-8-(3-dihydrobenzofuran-5-yl)-14-methyl-3,6,9,12,19,23-hexano-1,4,7,10,13,18-hexaazacyclic trimerose-22-yl)-2-acetamidohexamide (Compound 24)

[0342] Compound 24 was synthesized using compound int-2a (2.4 g) as the starting material, following the synthetic procedure in Example 1. The third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid was replaced with compound 3i to give compound 24, a white solid (78 mg, yield: 2.3%). 1 H NMR(500MHz,DMSO-d6)δ14.08(d,2H),10.76(s,1H),8.95(s,1H),8.47(d,1 H),8.22(t,2H),8.06(d,1H),7.97(d,1H),7.73(d,1H),7.60(d,1H),7.50( dd,2H),7.39(d,1H),7.34(d,1H),7.14(s,1H),7.10–7.04(m,2H),7.02–6. 95(m,2H),6.91(d,1H),6.63(d,1H),4.65(dd,1H),4.47(t,2H),4.43–4.35 (m,2H),4.26–4.21(m,1H),4.17(dd,2H),3.63(s,2H),3.08(dt,4H),2.99( dd,3H),2.96–2.86(m,3H),2.81(dd,1H),2.70(dd,1H),2.01(dd,2H),1.88 (d,1H),1.85(s,3H),1.81(d,1H),1.67(s,1H),1.56(s,1H),1.43(dd,1H), 1.34(d,2H),1.24(s,12H),1.02(d,3H),0.82(t,3H)ppm; LCMS: 1037.6[M+1] + .

[0343] Example 25

[0344] (S)-N-((2S,5R,8S,11S,14S,22S)-2-((1H-imidazol-4-yl)methyl)-11-((1H-indol-3-yl)methyl)-5-((2,3-dihydrobenzofuran-5-yl)methyl)-8-(3-dihydrobenzofuran-5-yl)methyl)-8-(3-guanidinopropyl)-14-methyl-3,6,9,12,19,23-hexano-1,4,7,10,13,18-hexaazacyclic trimerose-22-yl)-2-acetamidohexamide (Compound 25)

[0345] Compound 25 was synthesized using compound int-2b (2.4 g) as the starting material, following the synthetic procedure of Example 1. The third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid was replaced with compound 3i to give compound 25, a white solid (63 mg, yield: 2.1%). 1 H NMR(500MHz,DMSO-d6)δ14.01(s,1H),10.83(s,1H),8.91(s,1H),8.43(d,1H),8.31(d,1H),8.16(d,1H),7.98(d,2H),7.92(d,1H) ,7.79(d,1H),7.58(d,1H),7.54-7.47(m,2H),7.33(d,2H),7.13(s,1H),7.09-7.01(m,3H),6.98(t,1H),6.88(d,2H),6.62(d,1H), 4.65–4.53(m,1H),4.52–4.39(m,4H),4.26–4.08(m,3H),3.68(s,1H),3.24(s,1H),3.04(M,6H),2.97–2.81(m,3H),2.78–2.58(m,2 H),1.98(t,2H),1.84(s,4H),1.77(d,1H),1.59(d,2H),1.44(d,2H),1.38–1.17(m,12H),0.89–0.77(m,6H)ppm; LCMS: 1037.6[M+1] + .

[0346] Example 26

[0347] (S)-N-((6S,9S,12R,15S,18S)-15-((1H-imidazol-4-yl)methyl)-6-((1H-indol-3-yl)methyl)-12-((2,3-dihydrobenzofuran-5-yl)methyl)-9-(3-guanidinopropyl)-5,8,11,14,17,21-hexano-4,7,10,13,16,22-hexaazapyran[2.22]pentahydroxy-18-yl)-2-acetaminohexanamide (compound 26)

[0348] Compound 26 was synthesized using compound 20f (2.4 g) as the starting material, following the synthetic procedure of Example 1. The third amino acid (R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(4-fluorophenyl)propionic acid was replaced with compound 3i to give compound 26, a white solid (73 mg, yield: 2.1%). 1 H NMR(500MHz,DMSO-d6)δ14.10(d,2H),10.80(s,1H),8.94(s,1H),8.49(d,1H),8 .34-8.14(m,2H),8.07(s,1H),7.98(d,2H),7.81(d,1H),7.64–7.46(m,3H),7.3 3(d,1H),7.13–7.01(m,3H),6.97(t,2H),6.90(d,1H),6.62(d,1H),4.70-4.59( m,1H),4.52–4.42(m,3H),4.42–4.34(m,1H),4.29–4.09(m,3H),3.37–3.16(m,2 H),3.14–3.00(m,5H),2.99–2.82(m,4H),2.81–2.72(m,1H),2.70–2.62(m,1H), 2.00(d,2H),1.94–1.79(m,4H),1.79–1.71(m,1H),1.70–1.61(m,2H),1.60–1.5 2(m,1H),1.51–1.41(m,2H),1.41–1.28(m,4H),1.28–1.12(m,6H),1.04(s,1H), 0.89–0.78(m,3H),0.52–0.43(m,1H),0.43–0.21(m,2H)ppm; LCMS: 1049.6[M+1] + .

[0349] Biological testing

[0350] Example 1: Compound promotes cAMP production

[0351] This embodiment uses the LANCE Ultra cAMP kit (Revvity, TRF0264) to detect cellular cAMP levels, aiming to analyze the agonistic effects of compounds on MC1R and MC5R in cells. CHO cells overexpressing MC1R and MC5R were cultured in F-12 medium (37°C, 5% CO2) containing 10% fetal bovine serum and 0.2 mg / mL Hygromycin B. Cells were passaged at approximately 80% confluence or subjected to the following experiments. A 1×Stimulation Buffer was prepared according to the LANCE Ultra cAMP kit instructions. The test compounds were serially diluted 10× using the 1×Stimulation Buffer (MC1R starting at 1 nM, MC5R starting at 10 nM; 1:3 dilution, 10 concentrations). Count and dilute the cells obtained from the digestion and resuspending process to the target concentrations (55,000 cells / mL for MC1R and 220,000 cells / mL for MC5R). Add 9 μL of cells to each well of a 384-well plate (500 cells / well for MC1R and 2,000 cells / well for MC5R). Then add 1 μL of the corresponding concentration of compound to each well. Incubate at 37°C for 30 min. Dilute Eu-cAMP to the working concentration with detection buffer and add 5 μL to each well. Use detection buffer to... TM The anti-cAMP antibody was diluted to the working concentration, and 5 μL was added to each well; incubated at room temperature for 1 h. The emission values ​​at 665 nm and 620 nm were detected using a microplate reader (BMG, PHERAstar FSX) under excitation at 330 nm, and the 665 / 620 ratio was calculated. The cAMP production at each concentration was calculated, and curve fitting and EC50 were performed using GraphPad with a 4-parameter method. 50 The calculations are shown in Table 1.

[0352] Table 1

[0353] Example 2: Compound inhibits TNF-α production in mice

[0354] The inhibitory effect of the compound on this pro-inflammatory factor was evaluated by detecting the TNF-α content in mice. Thirty male C57 / 6j mice (5 mice / group) aged 4-6 weeks (Vitollivar) were injected via tail vein with 0.5 mg / kg of the test compound (dissolved in physiological saline, 5 mL / kg); 30 min later, each mouse was injected intraperitoneally with 5 μg LPS (Sigma, L2630); 1 h later, 50 μL of blood was collected via the orbital venous plexus. After the blood was left at room temperature for 30 min, it was centrifuged (4000g, 10 min) to obtain serum. The serum TNF-α content was then detected using a Mouse TNF-α Elisa kit (Invitrogen, ECM008). The TNF-α inhibition rate (%) was calculated using a control group (no compound administered, only LPS induction), as shown in Table 2.

[0355] Table 2

[0356] It will be apparent to those skilled in the art that the present invention is not limited to the foregoing illustrative embodiments, but may be embodied in other specific forms without departing from its essential characteristics. Therefore, the embodiments are intended to be illustrative and non-limiting in all respects, and reference should be made to the appended claims rather than to the foregoing embodiments; thus, all variations within the meaning and scope of the equivalents of the appended claims are included herein.

Claims

1. A cyclic peptide compound having the structure shown in Formula (I), or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of Formula (I), in, R 1 It can be hydrogen, deuterium, alkyl, haloalkyl, heteroalkyl, or cycloalkyl; R 2 It can be hydrogen, deuterium, alkyl, heterocyclic, or heteroaryl; R 3 and R 4 Each is independently an aryl or heteroaryl group; R 5 It can be amino, alkyl, alkoxy, heteroaryl, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3, -S(=O)-CH3 or -C(=O)-OR 9 ; R 6 Aryl, heteroaryl, heterocyclic, or cycloalkyl; R 7 and R 8 Each of these can be independently hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, alkyl, alkenyl, alkynyl, haloalkyl, heteroalkyl, alkoxy, hydroxyalkyl, alkoxyalkyl, alkenylalkyl, cycloalkylalkyl, haloalkoxy, cycloalkyloxy, heterocyclicoxy, cycloalkyl, heterocyclic, aryl, or heteroaryl. Optionally, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles; R 9 It can be hydrogen, alkyl, haloalkyl, deuterated alkyl, hydroxyalkyl, or cycloalkyl; R 10 It can be hydrogen, deuterium, halogen, hydroxyl, alkyl, haloalkyl, deuterated alkyl, hydroxyalkyl, or cycloalkyl; A is -(CH2) p -or-(CH2) q -O-(CH2) u -; m, n, t, p, q or u are each independently 0, 1, 2, 3, 4 or 5; R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 The groups described in A are independently and optionally replaced by 1, 2, 3 or 4 substituents selected from deuterium, halogen, amino, hydroxyl, cyano, oxo, nitro, alkyl, alkoxy, alkenyl, alkynyl, haloalkyl, cyanoalkyl, alkoxyalkyl, cycloalkylalkyl, heteroalkyl, heteroalkylalkyl, alkenylalkyl, alkynylalkyl, aryl, heteroaryl, heterocyclic, and cycloalkyl.

2. The cyclic peptide compound according to claim 1, wherein, R 5 For amino, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-9 heteroaryl, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3, -S(=O)-CH3 or -C(=O)-OR 9 ; R 6 C 6-10 Aryl, C 1-9 heteroaryl, C 1-9 Heterocyclic group or C 3-8 cycloalkyl; R 9 For hydrogen, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Deuterated alkyl, C 1-6 Hydroxyalkyl or C 3-8 Cycloalkyl.

3. The cyclic peptide compound according to claim 2, wherein, R 5 It is an amino group, -NH-(CH2)-NH2, -NH-C(=NH)-NH2, -NH-C(=O)-NH2, -S(=O)2CH3 or -S(=O)-CH3; R 6 C 6-10 Aryl or C 1-9 heteroaryl; preferably 4. The cyclic peptide compound according to any one of claims 1-3, wherein, The compound has a structure as shown in formula (II), or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug of a compound with a structure as shown in formula (II). Among them, R 1 R 2 R 3 R 4 R 7 R 8 A, m, and n are defined in equation (I).

5. The cyclic peptide compound according to claim 1 or 4, wherein, in, R 1 For hydrogen, deuterium, C 1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Heteroalkyl or C 3-8 cycloalkyl; R 2 For hydrogen, deuterium, C 1-6 Alkyl, C 1-9 Heterocyclic groups or independently optionally halogenated, C 1-4 Alkyl-substituted C 1-9 Mixed aromatic compounds.

6. The cyclic peptide compound according to claim 5, wherein, in, R 1 C is methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, or optionally substituted with 1, 2, 3, 4, 5, or 6 F, Cl, Br, or OH groups. 1-4 alkyl; R 2 C 1-4 Alkyl group, independently and optionally with 1, 2, or 3 halogens or with C 1-4 Alkyl-substituted pyrrole, pyrazol, imidazole, or triazolyl groups.

7. The cyclic peptide compound according to claim 1 or 4, wherein, R 3 and R 4 Each independently can be halogenated or C 1-4 Alkyl-substituted C 6-10 Aryl or C 1-9 Mixed aromatic compounds.

8. The cyclic peptide compound according to claim 7, wherein, R 3 and R 4 Each can be independently selected by 1, 2, or 3 F, Cl, Br, or C atoms. 1-4 Alkyl-substituted pyrrole, pyrazolyl, imidazolyl, triazolyl, phenyl, naphthyl, pyridyl, or C 5-9 Dense heteroaryl compounds.

9. The cyclic peptide compound according to claim 1 or 4, wherein, R 7 and R 8 Each of these can be independently represented by hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, or C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Haloalkyl, C 1-6 Heteroalkyl, C 1-6 Alkoxy, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy C 1-6 Alkyl, C 2- 6-Alkenyl C 1-6 Alkyl, C 3-6 cycloalkyl C 1-6 Alkyl, C 1-6 Halogenated alkoxy groups, C 3-6 Cycloalkyloxy, C 1-6 Heterocyclic oxygen, C 3-6 cycloalkyl, C 1-6 Heterocyclic group, C 6-10 Aryl or C 1-6 Mixed aromatics; Optionally, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles; R 10 Hydrogen, deuterium, halogen, hydroxyl, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Deuterated alkyl, C 1-3 Hydroxyalkyl or C 3- 6-Cycloalkyl.

10. The cyclic peptide compound according to claim 9, wherein, R 7 and R 8 Each of these can be independently represented by hydrogen, deuterium, halogen, hydroxyl, amino, cyano, nitro, or C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Heteroalkyl, C 1-3 Alkoxy, C 1-3 Hydroxyalkyl or C 1-3 Alkoxy C 1-3 alkyl; Optionally, R 7 R 8 Together with the carbon atom attached thereto, they form a group optionally bounded by 1, 2, 3 or 4 R atoms. 10 The substituted 3-7 membered carbon rings or optionally replaced by 1, 2, 3 or 4 Rs 10 Substituted 3-7 membered heterocycles; R 10 For hydrogen, deuterium, halogen, hydroxyl, methyl, ethyl, C substituted with 1, 2, or 3 F or Cl atoms 1-3 Alkyl, C 1- 3-Deuterated alkyl, C 1-3 Hydroxyalkyl or C 3-6 Cycloalkyl.

11. The cyclic peptide compound according to any one of claims 1-10, wherein, The compound has one of the following structures, or a stereoisomer, tautomer, nitride, solvate, metabolite, pharmaceutically acceptable salt, or prodrug, or a thereof.

12. A pharmaceutical composition comprising the cyclic peptide compound of any one of claims 1-11, and a pharmaceutically acceptable carrier, excipient, diluent, adjuvant, medium, or combination thereof.

13. Use of the cyclic peptide compound of any one of claims 1-11 or the pharmaceutical composition of claim 12 in the preparation of a medicament for the prevention, treatment, relief or regulation of diseases mediated by melanocortin receptors.

14. The use according to claim 13, wherein the disease is an ocular disease, selected from one or more of the following: dry eye syndrome, corneal ulcer, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scar, epithelial defect, keratoconjunctivitis, idiopathic uveitis, corneal transplantation, age-related macular degeneration, diabetic retinopathy, blepharitis, glaucoma, ocular hypertension, postoperative eye pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal neovascularization. Ocular lesions, vascular occlusive diseases, allergic eye diseases, tumors, retinitis pigmentosa, ocular infections, scleritis, ptosis, miosis, eye pain, mydriasis, neuralgia, cicatricial ocular surface diseases, ocular infections, inflammatory eye diseases, ocular surface diseases, corneal diseases, retinal diseases, ocular manifestations of systemic diseases, hereditary ocular diseases, ocular tumors, increased intraocular pressure, herpes infections, pterygium, wounds extending to the ocular surface, post-refractive keratotomy ocular pain and inflammation, corneal thermal or chemical burns, scleral wounds, keratoconus or conjunctival wounds.

15. A method for treating a disease, indication, symptom, or syndrome mediated by melanocortin receptor in a human or non-human mammal, the method comprising administering the cyclic peptide compound of any one of claims 1-11 or the pharmaceutical composition of claim 12.

16. The method of claim 15, wherein the melanocortin-mediated disease is an ocular disease selected from one or more of the following: dry eye, corneal ulcer, corneal erosion, corneal abrasion, corneal degeneration, corneal perforation, corneal scarring, epithelial defects, keratoconjunctivitis, idiopathic uveitis, corneal transplantation, age-related macular degeneration, diabetic retinopathy, blepharitis, glaucoma, ocular hypertension, postoperative ocular pain and inflammation, posterior segment neovascularization, proliferative vitreoretinopathy, cytomegalovirus retinitis, endophthalmitis, choroidal... Neovascular membranes, vascular occlusive diseases, allergic eye diseases, tumors, retinitis pigmentosa, ocular infections, scleritis, ptosis, miosis, eye pain, mydriasis, neuralgia, cicatricial ocular surface diseases, ocular infections, inflammatory eye diseases, ocular surface diseases, corneal diseases, retinal diseases, ocular manifestations of systemic diseases, hereditary ocular diseases, ocular tumors, elevated intraocular pressure, herpes infections, pterygium, wounds extending to the ocular surface, post-refractive keratotomy ocular pain and inflammation, corneal thermal or chemical burns, scleral wounds, keratoconus or conjunctival wounds.

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