Glycopeptide antibiotic-amino / guanidino derivative, and preparation method, pharmaceutical composition and application thereof

Abstract

The invention belongs to the technical field of medicinal chemistry and medicine, and particularly relates to a glycopeptide antibiotic-amino / guanidino derivative, pharmaceutically acceptable salt thereof, a preparation method thereof, a pharmaceutical composition and application of the glycopeptide antibiotic-amino / guanidino derivative. The glycopeptide antibiotic-amino / guanidino derivative, especially a norvancomycin-amino / guanidino derivative or vancomycin-amino / guanidino derivative, has a structure as shown in formula (I). Different parts of norvancomycin and vancomycin are modified, and extra terminal primary amino or guanidino is introduced, so that the glycopeptide antibiotic-amino / guanidino derivative is obtained. The invention further explores the specific action mechanisms of the Gram-positive bacteria and the Gram-negative bacteria, so that a new variety is added for the antibiotic Wushu for resisting the drug resistance of the glycopeptide antibiotics, and the Gram-positive bacteria and the Gram-negative bacteria can be used for resisting the drug resistance of the glycopeptide antibiotics by using the Gram-positive bacteria and the Gram-negative bacteria as well as the specific action mechanisms of the Gram-positive bacteria and the Gram-negative bacteria.

Description

A class of glycopeptide antibiotics - amino / guanidinyl derivatives, their preparation methods, pharmaceutical compositions and uses Technical Field

[0001] This invention belongs to the fields of medicinal chemistry and pharmaceutical technology, specifically relating to a class of glycopeptide antibiotics—amino / guanidinyl derivatives, particularly vancomycin or norvancomycin derivatives with a primary amino or guanidinyl group at the end of the modified fragment, and pharmaceutically acceptable salts thereof; a method for preparing the glycopeptide antibiotics—amino / guanidinyl derivatives; a pharmaceutical composition comprising the glycopeptide antibiotics—amino / guanidinyl derivatives or pharmaceutically acceptable salts thereof; and the use of the glycopeptide antibiotics—amino / guanidinyl derivatives or pharmaceutically acceptable salts thereof, or the pharmaceutical composition thereof, in the preparation of a medicament for treating and / or preventing diseases or conditions related to Gram-positive and / or Gram-negative bacterial infections. Background Technology

[0002] Antimicrobial resistance (AMR), a slowly evolving pandemic, was declared one of the top ten global public health threats by the World Health Organization (WHO) in 2019. AMR leads to serious infections with high morbidity and mortality rates. Drug-resistant Gram-negative pathogens, such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacteriaceae, have been a major concern, posing the greatest threat to human health. Whether in the well-known "ESKAPE" strain, the 2019 US Antibiotic Resistance Threat Report, or the WHO's 2017 and 2024 Bacterial Priority Pathogens (BPPL) lists, there is an urgent need for new antibiotics. Although there were 76 antimicrobial candidates in clinical trials as of 2021, relatively few are in late-stage clinical development with clinical differentiation, especially against key priority pathogens. Therefore, there is an urgent need to develop new antibiotics, particularly those targeting key priority pathogens, to address the current pressing situation of antibiotic resistance.

[0003] Vancomycin and Norvancomycin both belong to the glycopeptide antibiotic family and are considered the last line of defense against drug-resistant positive bacterial infections. Vancomycin was isolated by Eli Lilly scientists in the 1950s from the fermentation broth of *Amycosis orientalis* and approved for marketing in 1958, and has been used clinically for over 60 years. Norvancomycin was isolated by Chinese scientists in 1959 from the fermentation of *Actinomyces wan 23*, which was isolated from soil in Guizhou, and began mass production for clinical use in China in 1967. Currently, it is only available in China. Structurally, the difference between Norvancomycin and Vancomycin lies in the methyl group on the α-amino group of leucine at position 1. Their mechanisms of action are identical: they hinder subsequent transglycosylation and transpeptidation by forming five hydrogen bonds with the terminal dipeptide of the peptidoglycan precursor Lipid II and by the hydrophobic interaction of the aromatic group cavity in its parent nucleus. Ultimately, this prevents peptidoglycan from forming a mature network structure to protect bacterial cells, making the cells unable to withstand changes in osmotic pressure and resulting in lysis and death. However, with the first case of vancomycin-resistant enterococci (VRE) reported in France in 1986, followed by the first case of moderately vancomycin-resistant Staphylococcus aureus (VISA) reported in Japan in 1996, and the first report of highly vancomycin-resistant Staphylococcus aureus (VRSA) in 2002, the "last line of defense" is in grave danger. The emergence of more and more vancomycin-resistant strains poses a significant challenge and threat once again. Therefore, developing effective strategies against vancomycin-resistant strains is urgently needed.

[0004] The unique structure of Gram-negative bacteria, with their peptidoglycan layer located in the periplasm between the outer and inner membranes, makes them naturally resistant to glycopeptide antibiotics like vancomycin and teicoplanin. This has led to consideration of whether glycopeptide antibiotics can be transformed into effective antibiotics against drug-resistant Gram-negative bacteria, overcoming their inherent resistance. Given their unique mechanism of action, the absence of significant cross-resistance, and the long development time required for resistance mechanisms to take hold, developing novel glycopeptide antibiotics against drug-resistant Gram-negative bacteria, using first-generation glycopeptides such as vancomycin and the domestically unique teicoplanin as lead compounds, holds immense potential and significance.

[0005] Modification of small-molecule antibacterial compounds with terminal primary amino and guanidinyl groups, under physiological conditions, facilitates additional interactions with bacterial cell membranes due to their additional positive charge, thereby enhancing antibacterial activity. Previous studies have shown that terminal guanidinyl modification at the C-terminus of vancomycin can synergistically work with (4-chlorobiphenyl)methyl (CBP) modification at the vancomycin glycosamine site to combat drug-resistant Gram-positive bacteria (ACS Infect. Dis. 2020, 6(8), 2169-2180); single arginine modification at the C-terminus of vancomycin can overcome the inherent resistance of Gram-negative bacteria and make it effective against drug-resistant Escherichia coli (ACS Chem. Biol. 2019, 14(9), 2065-2070); while small molecules with molecular weights less than 500D typically enhance their antibacterial activity by increasing their penetration into bacterial cell membranes through additional amination modifications (Ann. NY Acad. Sci. 2019, 1435(1), 18-38).

[0006] Based on the above research, we found that the diversity of guanidine-containing fragments in these studies is limited, and the potential of this novel strategy has not been fully elucidated. Previous studies have not investigated modifications at sites other than the C-terminus of vancomycin, and the additional mechanisms of action behind them require further clarification. Furthermore, the impact of a single amino group on the complex molecular structure of vancomycin (molecular weight > 1000D) remains unexplored. More importantly, research on aminoation / guanidinolation of the core structure of norvancomycin, a glycopeptide antibiotic unique to China, is lacking. Summary of the Invention

[0007] This invention addresses the shortcomings of the existing technology by providing a class of glycopeptide antibiotics—amino / guanidinyl derivatives, their preparation methods, pharmaceutical compositions, and uses. By modifying different sites of vancomycin and desmethylvancomycin to introduce additional terminal primary amino or guanidinyl groups, the invention explores their effects on the antibacterial activity of acquiredly resistant Gram-positive bacteria and intrinsically resistant Gram-negative bacteria, and further investigates their specific mechanisms of action, thus adding new varieties to the antibiotic arsenal against glycopeptide antibiotic resistance.

[0008] The specific technical solution is as follows:

[0009] The first objective of this invention is to provide a class of glycopeptide antibiotics—amino / guanidinyl derivatives, particularly vancomycin-amino / guanidinyl derivatives or vancomycin-amino / guanidinyl derivatives, having the structure shown in formula (I):

[0010]

[0011] in:

[0012] R0 is selected from -H or -CH3;

[0013] R1 is selected from -OH, -NH-L-NH2, and -NH-L-guanidinyl groups;

[0014] R2 is selected from -H, -L-NH2, and -L-guanidino groups;

[0015] R3 is selected from -H, -CH2-NH-L-NH2, and -CH2-NH-L-guanidinyl groups;

[0016] R4 is selected from -L-NH2 or -L-guanidinyl groups;

[0017] Furthermore, at least one of R1, R2, R3, and R4 contains a -NH2 or -guanidinyl structural fragment;

[0018] L, as a linker, is independently selected from the following groups:

[0019] Substituted or unsubstituted C4-C 20 Straight-chain or branched alkylene, substituted or unsubstituted C4-C 20 Straight-chain or branched alkenyl groups, substituted or unsubstituted C4-C 20 Straight-chain or branched ynylene groups, C4-C 20 Straight-chain or branched imide groups, substituted or unsubstituted C3-C 10 Cycloalkylene, substituted or unsubstituted C6-C 20 3-10 membered nonaromatic heterocyclic groups containing one or more heteroatoms selected from N, O and S on a arylene ring, substituted or unsubstituted ring; 5-10 membered heteroarylene groups containing one or more heteroatoms selected from N, O and S on a substituted or unsubstituted ring.

[0020] The substitution refers to substitution by one or more substituents selected from the following: halogen, -O-, -OH, -NH2, carbonyl, cyano, C1-C. 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Alkylamino, C1-C 10 Alkyl group, C3-C 10 cycloalkyl, halogenated C1-C 10 Alkyl, C2-C6 straight-chain or branched alkynyl, phenylethynyl, trimethylsilylethynyl, pyridyl, phenyl, cyanophenyl, C1-C6 alkylphenyl, trifluoromethylphenyl, chlorophenyl, biphenyl, methyl biphenyl, trifluoromethyl biphenyl, halogen-substituted biphenyl.

[0021] Furthermore, L, as a linker, is independently selected from the following groups:

[0022] Substituted or unsubstituted C4-C 10Straight-chain or branched alkylene, substituted or unsubstituted C4-C 10 Straight-chain or branched alkenyl groups, substituted or unsubstituted C4-C 10 Straight-chain or branched ynylene groups, C4-C 10 Straight-chain or branched imide groups, substituted or unsubstituted C3-C 10 Cycloalkylene, substituted or unsubstituted C6-C 14 3-10 membered nonaromatic heterocyclic groups containing one or more heteroatoms selected from N, O and S on a arylene ring, substituted or unsubstituted ring; 5-10 membered heteroarylene groups containing one or more heteroatoms selected from N, O and S on a substituted or unsubstituted ring.

[0023] The substitution refers to substitution by one or more substituents selected from the following: halogen, -O-, -OH, -NH2, carbonyl, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkylacyl, C3-C 10 Cycloalkyl, halogenated C1-C6 alkyl, C2-C6 straight-chain or branched alkynyl, phenylethynyl, trimethylsilylethynyl, pyridyl, phenyl, cyanophenyl, C1-C6 alkylphenyl, trifluoromethylphenyl, chlorophenyl, biphenyl, methyl biphenyl, trifluoromethyl biphenyl, halogen-substituted biphenyl.

[0024] Furthermore, L, as a linker, is independently selected from the following groups:

[0025] -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7-, -(CH2)8-, -CO(CH2)2-, -CO(CH2)3-, -(CH2)2-O-(CH2)2-, -(CH2)2-O-(CH2)2-O-(CH2)2-, -CH2-benzene-CH2-, -CH2-biphenyl-CH2-.

[0026] Furthermore, the vancomycin-amino / guanidinyl derivative or vancomycin-amino / guanidinyl derivative shown in formula (I) is selected from the following compounds:

[0027]

[0028]

[0029]

[0030]

[0031]

[0032] A second object of the present invention is to provide a pharmaceutically acceptable salt of a glycopeptide antibiotic-amino / guanidine derivative as described above, particularly a pharmaceutically acceptable salt of a vancomycin-amino / guanidine derivative or a vancomycin-amino / guanidine derivative as described above.

[0033] A third objective of this invention is to provide a method for preparing the glycopeptide antibiotic-amino / guanidinyl derivatives as described above, particularly a method for preparing the vancomycin-amino / guanidinyl derivatives or vancomycin-amino / guanidinyl derivatives as described above, comprising the following steps: vancomycin hydrochloride or vancomycin hydrochloride undergoes a site-selective amide condensation reaction, a Mannich reaction, or a reductive amination reaction, and a deprotection reaction to obtain vancomycin or vancomycin derivatives with a terminal primary amino group; a guanidinylation reaction is then carried out, and a vancomycin derivative or vancomycin derivative with a terminal guanidinyl group is obtained through a selective substitution reaction and an excess of a guanidinylating agent.

[0034] Furthermore, the amide condensation reaction, Mannich reaction, or reductive amination reaction is carried out at -20°C to 70°C; wherein, the amide condensation reaction temperature is preferably 37°C, the Mannich reaction temperature is preferably -10°C, the reductive amination reaction at the R3 site is preferably 50°C, and the reductive amination reaction at the R4 site is preferably 70°C.

[0035] Furthermore, the deprotection reaction is carried out at -10℃ to 10℃, preferably 0℃.

[0036] Furthermore, the solvent for the deprotection reaction is a trifluoroacetic acid-water mixture or a trifluoroacetic acid-dichloromethane mixture, preferably a trifluoroacetic acid-dichloromethane mixture.

[0037] Furthermore, the volume content of trifluoroacetic acid in the solvent is 10% to 30%, preferably 20%.

[0038] Furthermore, the deprotection reaction takes 10 min to 2 h, preferably 30 min.

[0039] Furthermore, the guanidinizing agent is selected from 1H-pyrazole-1-formamidine hydrochloride, N-BOC-1H-pyrazole-1-formamidine, N,N'-di-BOC-1H-1-guanidinylpyrazole, N,N'-di-BOC-S-methylisothiourea, preferably 1H-pyrazole-1-formamidine hydrochloride.

[0040] Furthermore, the guanidination reaction is carried out at 20°C to 70°C, preferably 60°C or 70°C.

[0041] Specifically, the preparation method of the vancomycin-amino / guanidinyl derivative or the vancomycin-amino / guanidinyl derivative can be one of the following four preparation methods:

[0042] Method 1:

[0043] Norvancomycin hydrochloride or vancomycin hydrochloride reacts with the NHBoc-L-NH2 fragment via an amide condensation reaction and a deprotection reaction to yield the corresponding norvancomycin-amino derivative or vancomycin-amino derivative of formula (I); further, this derivative undergoes a guanidinolation reaction to yield the corresponding norvancomycin-guanidino derivative or vancomycin-guanidino derivative of formula (I):

[0044]

[0045] in,

[0046] The definition of L is the same as that in equation (I) above.

[0047] Method 2:

[0048] Norvancomycin hydrochloride or vancomycin hydrochloride reacts with the NHBoc-L-CHO fragment via a reductive amination reaction and a deprotection reaction to yield the corresponding norvancomycin-amino derivative or vancomycin-amino derivative of formula (I); further, this derivative undergoes a guanidinolation reaction to yield the corresponding norvancomycin-guanidino derivative or vancomycin-guanidino derivative of formula (I):

[0049]

[0050] in,

[0051] The definition of L is the same as that in equation (I) above.

[0052] Method 3:

[0053] Norvancomycin hydrochloride or vancomycin hydrochloride reacts with the NHBoc-L-NH2 fragment via a Mannich reaction and a deprotection reaction to yield the corresponding norvancomycin-amino derivative or vancomycin-amino derivative of formula (I); further, this derivative undergoes a guanidinolation reaction to yield the corresponding norvancomycin-guanidino derivative or vancomycin-guanidino derivative of formula (I):

[0054]

[0055] in,

[0056] The definition of L is the same as that in equation (I) above.

[0057] Method 4:

[0058] Norvancomycin hydrochloride or vancomycin hydrochloride reacts with the NHBoc-L-CHO fragment via a reductive amination reaction and a deprotection reaction to yield the corresponding norvancomycin-amino derivative or vancomycin-amino derivative of formula (I); further, this derivative undergoes a guanidinolation reaction to yield the corresponding norvancomycin-guanidino derivative or vancomycin-guanidino derivative of formula (I):

[0059]

[0060] in,

[0061] The definition of L is the same as that in equation (I) above.

[0062] A fourth object of the present invention is to provide a pharmaceutical composition comprising the glycopeptide antibiotics as described above—amino / guanidine derivatives or pharmaceutically acceptable salts thereof; particularly comprising vancomycin—amino / guanidine derivatives or pharmaceutically acceptable salts thereof, or vancomycin—amino / guanidine derivatives or pharmaceutically acceptable salts thereof.

[0063] A fourth object of the present invention is to provide the use of the glycopeptide antibiotics as described above—amino / guanidinyl derivatives or pharmaceutically acceptable salts thereof, or pharmaceutical compositions as described above, in the preparation of a medicament for treating and / or preventing diseases or conditions associated with Gram-positive and / or Gram-negative bacterial infections; particularly the use of vancomycin as described above—amino / guanidinyl derivatives or pharmaceutically acceptable salts thereof, vancomycin as described above—amino / guanidinyl derivatives or pharmaceutically acceptable salts thereof, or pharmaceutical compositions as described above, in the preparation of a medicament for treating and / or preventing diseases or conditions associated with Gram-positive and / or Gram-negative bacterial infections.

[0064] Furthermore, the Gram-positive bacteria include, but are not limited to, Staphylococcus aureus and Enterococcus.

[0065] Furthermore, the Gram-negative bacteria include, but are not limited to: Escherichia coli, Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa.

[0066] Furthermore, the diseases or conditions associated with Gram-positive and / or Gram-negative bacterial infections include, but are not limited to: respiratory tract infections (upper respiratory tract infections, such as pharyngitis; lower respiratory tract infections including tracheitis, bronchitis, pneumonia caused by Enterobacter spp. and Serratia marcescens, such as community-acquired pneumonia, ventilator-associated pneumonia, hospital-acquired pneumonia, bronchiectasis), pulmonary tuberculosis and pulmonary infections complicated by pulmonary fibrosis, urinary tract infections (including uncomplicated and complicated pyelonephritis, recurrent cystitis, complicated urethritis) Infections include: urinary tract infections, central nervous system infections (encephalitis, meningitis, brain abscess), ear infections (otitis externa, otitis media), abdominal infections (including peritonitis), cardiovascular infections (bloodstream infections, such as sepsis or bacteremia, endocarditis, myocarditis, pericarditis), skin or soft tissue infections, bone and joint infections (arthritis, osteomyelitis), genital infections (genital ulcers, vaginitis, cervicitis), eye infections (conjunctivitis, keratitis, endophthalmitis), and oral infections (including gingivitis, periodontitis).

[0067] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0068] (1) This invention explores the effects of modified vancomycin and different parts of vancomycin on the antibacterial activity of acquired drug-resistant Gram-positive bacteria and inherently drug-resistant Gram-negative bacteria by introducing additional terminal primary amino or guanidine groups, and further explores their specific mechanisms of action, adding new varieties to the antibiotic arsenal to combat glycopeptide antibiotic resistance.

[0069] (2) The antibacterial experiments of the present invention show that the structural modification strategy of the novel vancomycin-amino / guanidinyl derivatives and vancomycin-amino / guanidinyl derivatives involved in the present invention can significantly enhance their antibacterial activity and show potential against the inherent drug resistance of Gram-negative bacteria.

[0070] (3) The cytotoxicity experiment of the present invention shows that the compound prepared in the present invention has no cytotoxicity to the HEK-293T cell line at the measured concentration and has high safety.

[0071] (4) The hemolytic toxicity test of the present invention shows that the compound NV014 prepared in the present invention has no hemolytic toxicity at a drug concentration of 100 mg / L and has good safety.

[0072] (5) The time-kill curve experiment of the present invention shows that the bactericidal performance of the compound NV014 prepared by the present invention is better than that of the negative control vancomycin;

[0073] (6) The pharmacokinetic experiments in mice of the present invention showed that the compound NV014 of the present invention had a longer half-life (T1 / 2) than vancomycin, and the area under the concentration-time curve (AUC) was 4-5 times larger than that of vancomycin. In terms of plasma clearance (CL), the compound NV014 of the present invention had a slower clearance rate than vancomycin, showing better drug-likeness parameters than the positive compound vancomycin.

[0074] (7) The in vivo efficacy experiment in mice showed that the compound NV014 of the present invention has significant in vivo protective effect against the systemic infection model of drug-resistant bacteria MRSA252, and its in vivo efficacy is far superior to vancomycin. Attached Figure Description

[0075] Figure 1 is a bar chart showing the cytotoxicity test of the compound of the present invention against HEK-293T cells in biological test 2;

[0076] Figure 2 is a graph showing the hemolytic toxicity test of the compound of the present invention in biological test 3;

[0077] Figure 3 shows the time-kill kinetic curves of the compounds of the present invention against Staphylococcus aureus MRSA252 strain (a) and Escherichia coli AB1157 strain (b) in biological test 4.

[0078] Figure 4 shows the in vivo pharmacokinetic plasma concentration-time curve of the compound of the present invention in mice in biotest 5;

[0079] Figure 5 shows the survival rate curves of mice in the Staphylococcus aureus MRSA252 strain model after single-dose administration (20 mg / kg, 10 mg / kg) in Biotest 6. Detailed Implementation

[0080] The principles and features of the present invention are described below with reference to examples. The examples are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0081] Terminology Explanation

[0082] In this invention, the term "C1-C" X "Alkyl" refers to a straight-chain or branched alkyl group having 1 to X carbon atoms in its main chain, such as "C1-C". 10 "Alkyl" refers to a straight-chain or branched alkyl group having 1 to 10 carbon atoms in its main chain, preferably C1-C6 alkyl or C1-C4 alkyl, examples of which include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isopentyl, hexyl, etc.

[0083] In this invention, the term "aryl" refers to an aromatic cyclic group that does not contain heteroatoms, such as phenyl or naphthyl.

[0084] In this invention, the term "heteroaryl" refers to an aryl group containing one or more heteroatoms selected from N, O, and S, such as pyrroleyl, imidazolyl, oxazolyl, thiazolyl, furanyl, thiopheneyl, pyridinyl, pyrimidinyl, indolyl, quinolinyl, etc.

[0085] The term "pharmaceutically acceptable salt" in this invention refers to salts formed with inorganic acids such as phosphoric acid, sulfuric acid, and hydrochloric acid, or organic acids such as acetic acid, tartaric acid, citric acid, malic acid, and trifluoroacetic acid, or acidic amino acids such as aspartic acid and glutamic acid, or salts formed with the above acids as esters or amides and then with inorganic bases, such as sodium, potassium, calcium, aluminum, and ammonium salts.

[0086] For the following examples, standard operating and purification methods known to those skilled in the art were used. Unless otherwise specified, the raw materials were generally available from commercially available sources, such as Shanghai Titan Technology Co., Ltd. and Bidex Pharmaceutical Technology Co., Ltd. Commercially available solvents and reagents were generally used without further purification; anhydrous solvents were processed using standard methods, and other reagents were commercially available analytical grade. Specifically, vancomycin was purchased from Nanjing Zhongbiao Chenxi Chemical Technology Co., Ltd., batch number 130338-201704. Vancomycin was purchased from Titan Technology Co., Ltd., batch number 011073672. Vancomycin hydrochloride was purchased from Titan Technology Co., Ltd., batch number 011073672.

[0087] Unless otherwise stated, all temperatures are expressed in °C (degrees Celsius), room temperature or ambient temperature refers to 20–25 °C, and the structure of the compounds is determined by nuclear magnetic resonance (NMR) spectroscopy and / or mass spectrometry (MS).

[0088] The shifts (δ) in the proton NMR spectra are given in parts per million (ppm). Proton NMR spectra were determined using an AscEnd™-600MHz NMR spectra with deuterated methanol (CD3OD), deuterated chloroform (CDCl3), deuterated dimethyl sulfoxide (DMSO-d6), and deuterated water (D2O) as solvents, and tetramethylsilane (TMS) as an internal standard.

[0089] High-resolution mass spectrometry was performed using a Waters ACQUITY UPLC I-Class-Vion IMS QTof LC-MS. When describing the intensity of chlorine or bromine-containing ions, the expected intensity ratio was observed (approximately 3:1 for ions containing 35Cl / 37Cl and 1:1 for ions containing 79Br / 81Br), and only the intensity of the lower-mass ions was given.

[0090] HPLC: UltiMate 3000 analytical high-performance liquid chromatography system (Thermo Fisher Scientific). -3030 Analytical High Performance Liquid Chromatography System (Shanghai Tongwei Analytical Technology Co., Ltd.) and -3050 Preparative High Performance Liquid Chromatography System (Shanghai Tongwei Analytical Technology Co., Ltd.). Analytical High Performance Liquid Chromatography Conditions: C18 column (Welch...) 5 μm, 4.6 × 250 mm), UV detection bands were 214 nm and 254 nm, elution conditions were 2-45% acetonitrile (containing 0.1% v / v TFA) gradient wash for 20 min. Preparative high performance liquid chromatography conditions: C18 column (Welch) 5μm, 21.2×250mm), UV detection bands are 214nm and 254nm, elution conditions are 2-45% acetonitrile (containing 0.1% v / v TFA) gradient wash for 20 minutes.

[0091] In the foregoing and the following embodiments, the following abbreviations have the following meanings. If an abbreviation is not defined, it has the generally accepted meaning.

[0092] DCM is dichloromethane;

[0093] DIPEA is N,N-diisopropylethylamine;

[0094] DMF is N,N-dimethylformamide;

[0095] DMSO stands for dimethyl sulfoxide.

[0096] HATU is O-(pyridotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate;

[0097] MTBE is methyl tert-butyl ether;

[0098] TFA stands for trifluoroacetic acid;

[0099] TLC stands for Thin Layer Chromatography.

[0100] Preparation of starting materials

[0101] Preparation Example 1: Preparation of [({4-[4-(aminomethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (Compound 1)

[0102]

[0103] Weigh commercially available [4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl]methaneamine (233 mg, 1.0 mmol), {[(4-bromophenyl)methyl]amino}methane-2-methylpropyl-2-yl ester (285 mg, 1.0 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium dichloride (73 mg, 0.1 mmol), and potassium phosphate (637 mg, 3.0 mmol) into a thick-walled pressure-resistant tube. Add 4 mL of DME and 1 mL of water sequentially, and stir at 120 °C for 2 h. Detect the reaction by TLC until it stops. The reaction solution was then filtered with diatomaceous earth, and the filtrate was extracted three times with ethyl acetate. The organic layer was then washed successively with water, 1 mol / L HCl, and saturated brine. The organic layer was then separated, dried with anhydrous sodium sulfate, and purified by a rapid preparative column chromatography to obtain the target product compound 1 (240 mg, yield 77%).

[0104] Compound 1: 1 H NMR(400MHz,Chloroform-d)δ7.57-7.52(m,2H),7.47(d,J=6.2Hz,2H),7.38(d,J=8.0Hz,2H),7 .25(d,J=4.2Hz,2H),4.87(s,1H),4.37(d,J=6.0Hz,2H),3.91(s,2H),1.55(s,2H),1.46(s,9H).

[0105] Preparation Example 2: Preparation of [({4-[4-(2-aminoethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (compound 2)

[0106]

[0107] In Preparation Example 1, [4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl))phenyl]methaneamine was replaced with commercially available 2-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl]ethyl-1-amine. The remaining raw materials, reagents, and preparation steps were the same as in Preparation Example 1, yielding the target product compound 2 (243 mg, yield 74%).

[0108] Compound 2: 1H NMR(400MHz,Chloroform-d)δ7.51(dd,J=10.5,7.9Hz,4H),7.33(d,J=7.9Hz,2H),7.25(d,J=10.5Hz,2H), 4.92(s,1H),4.34(d,J=5.9Hz,2H),2.99(t,J=6.8Hz,2H),2.78(t,J=6.8Hz,2H),1.46(s,9H),1.34(s,2H).

[0109] Preparation Example 3: Preparation of [(2-{4-[4-(aminomethyl)phenyl]phenyl}ethyl)amino]methane-2-methylpropyl-2-yl ester (compound 3)

[0110]

[0111] In Preparation Example 1, {[(4-bromophenyl)methyl]amino}methane-2-methylpropyl-2-yl ester was replaced with commercially available {[2-(4-bromophenyl)ethyl]amino}methane-2-methylpropyl-2-yl ester. The remaining raw materials, reagents and preparation steps were the same as in Preparation Example 1, and the target product compound 3 (246 mg, yield 75%) was obtained.

[0112] Compound 3: 1 H NMR(600MHz,Chloroform-d)δ7.61-7.51(m,4H),7.42-7.36(m,2H),7.28-7.26(m,1H),7.26-7.24(m,1H) ,4.59(s,1H),3.92(s,2H),3.41(q,J=6.8Hz,2H),2.84(t,J=7.1Hz,2H),1.67-1.52(m,2H),1.44(s,9H).

[0113] Preparation Example 4: Preparation of [({3-[4-(aminomethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (compound 4)

[0114]

[0115] In Preparation Example 1, {[(4-bromophenyl)methyl]amino}methane-2-methylpropyl-2-yl ester was replaced with commercially available {[(3-bromophenyl)methyl]amino}methane-2-methylpropyl-2-yl ester. The other required raw materials, reagents and preparation methods were the same as in Preparation Example 1, and the target product compound 4 (170 mg, yield 54%) was obtained.

[0116] Compound 4:1 H NMR(600MHz,Chloroform-d)δ7.57-7.54(m,2H),7.51-7.47(m,2H),7.41-7.36(m,3H) ,7.27(s,1H),4.90(s,1H),4.40-4.36(m,2H),3.92(s,2H),1.94(s,2H),1.47(s,9H).

[0117] Preparation Example 5: Preparation of [({2-[4-(aminomethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (compound 5)

[0118]

[0119] In Preparation Example 1, {[(4-bromophenyl)methyl]amino}methane-2-methylpropyl-2-yl ester was replaced with commercially available {[(2-bromophenyl)methyl]amino}methane-2-methylpropyl-2-yl ester. The other required raw materials, reagents and preparation steps were the same as in Preparation Example 1, and the target product compound 5 (106 mg, yield 34%) was obtained.

[0120] Compound 5: 1 H NMR(600MHz,Chloroform-d)δ7.46(d,J=7.7Hz,1H),7.41(d,J=7.6Hz,2H),7.38-7.34(m,1H),7.34-7.30(m,1H),7. 29-7.26(m,2H),7.24-7.21(m,1H),4.72(t,J=5.8Hz,1H),4.30-4.25(m,2H),3.97(s,2H),3.13(s,2H),1.44(s,9H).

[0121] Preparation Example 6: Preparation of ({[4-(4-formylphenyl)phenyl]methyl}amino)methane-2-methylpropyl-2-yl ester (compound 6)

[0122]

[0123] In Preparation Example 1, [4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl]methaneamine was replaced with commercially available 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)phenyl-1-carboxaldehyde. The remaining raw materials, reagents, and preparation steps were the same as in Preparation Example 1, yielding the target product compound 6 (157 mg, yield 50%).

[0124] Compound 6: 1 H NMR (400MHz, DMSO-d6) δ10.05(s,1H),7.99(d,J=8.3Hz,2H),7.90(d,J=8.1Hz,2H),7.73(d, J=8.2Hz,2H),7.53-7.41(m,1H),7.37(d,J=7.9Hz,2H),4.18(d,J=6.1Hz,2H),1.40(s,9H).

[0125] Preparation Example 7: Preparation of ({2-[4-(4-formylphenyl)phenyl]ethyl}amino)methane-2-methylpropyl-2-yl ester (compound 7)

[0126]

[0127] In Preparation Example 6, {[(4-bromophenyl)methyl]amino}methane-2-methylpropyl-2-yl ester was replaced with commercially available {[2-(4-bromophenyl)ethyl]amino}methane-2-methylpropyl-2-yl ester. The remaining raw materials, reagents and preparation steps were the same as in Preparation Example 1, and the target product compound 7 (166 mg, yield 51%) was obtained.

[0128] Compound 7: 1 H NMR (400MHz, Methanol-d4) δ9.95 (s, 1H), 7.90 (d, J = 8.3Hz, 2H), 7.74 (d, J = 8.4Hz, 2H), 7.57 (d, J=8.2Hz,2H),7.28(d,J=8.2Hz,2H),3.25(t,J=7.1Hz,2H),2.77(t,J=7.3Hz,2H),1.38(s,9H).

[0129] Example 1: Preparation of compound NV001

[0130]

[0131] Weigh commercially available vancomycin hydrochloride (50 mg, 34 μmol) and [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester (8 mg, 50 μmol) into a 10 mL single-necked reaction flask. Add 1 mL DMF, 1 mL DMSO, and 18 μL DIPEA (102 μmol) sequentially, and stir vigorously until the reaction solution becomes clear. Then, add HATU (13 mg, 34 μmol) pre-dissolved in 250 μL DMF dropwise to the above system. Continue stirring at room temperature and monitor the reaction progress using analytical RP-HPLC until the reaction stops. Add TFA dropwise to adjust the reaction solution to a weakly acidic state to quench the reaction. Add methyl tert-butyl ether (MTBE) to precipitate a white precipitate, which is then centrifuged to obtain a white crude solid. Repeat this step three times, finally washing once with acetonitrile. Subsequently, the product was deprotected in DCM with 20% TFA at 0°C for 30 min. The resulting white crude solid was dissolved in an appropriate amount of water and acetonitrile, then separated and purified by preparative RP-HPLC. The target component was freeze-dried to obtain a white flocculent solid NV001 (34 mg, yield 68%). NV001 and subsequent final products were all trifluoroacetates.

[0132] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.75(s,1H),8.54-8.51(m,1H),8.23-8.15(m,1H),7.85(s,1H),7.59(d,J=8.4Hz,1H),7.54(s, 1H),7.49-7.45(m,1H),7.33(d,J=8.3Hz,1H),7.23-7.19(m,2H),6.77(d,J=8.4Hz,1H),6.71(d,J=8.5 Hz,1H),6.39(d,J=2.3Hz,1H),6.21(d,J=2.3Hz,1H),5.76(d,J=7.5Hz,1H),5.64(s,1H),5.28(d,J=7 .7Hz,1H),5.26-5.24(m,1H),5.23(s,1H),5.17-5.15(m,1H),5.13-5.10(m,1H),4.84-4.80(m,1H),4. 69-4.65(m,1H),4.48-4.44(m,1H),4.38(d,J=5.6Hz,1H),4.25-4.21(m,1H),4.12(s,1H),4.08-4.03 (m,1H),3.68(d,J=10.8Hz,1H),3.58-3.50(m,2H),3.41-3.37(m,2H),3.29-3.23(m,2H),3.18(s,1H), 2.92–2.87 (m, 2H), 2.68 (s, 1H), 2.16–2.09 (m, 1H), 1.91 (d, J = 12.4 Hz, 1H), 1.76–1.66 (m, 2H), 1.66–1.53 (m, 2H), 1.31 (s, 3H), 1.07 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.3 Hz, 3H), 0.90 (d, J = 6.3 Hz, 3H). High-resolution mass spectrometry (ESI) + C 67 H 79 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1476.4800, measured value is m / z 1476.4776.

[0133] Example 2: Preparation of compound NV002

[0134]

[0135] Weigh NV001 (30 mg, 20 μmol) and 1H-pyrazole-1-formamidinium hydrochloride (15 mg, 100 μmol) into a 10 mL single-necked reaction flask. Add 2 mL of DMF and 35 μL of DIPEA (200 μmol) sequentially. Stir vigorously until the reaction solution becomes clear. Transfer the reaction system to 70 °C and continue reacting until analytical RP-HPLC monitoring shows no further change in the reaction. Then, return the reaction system to room temperature and add TFA dropwise to adjust the reaction solution to a weakly acidic state to quench the reaction. Add methyl tert-butyl ether (MTBE) to precipitate a white precipitate, which is then centrifuged to obtain a white solid crude product. Repeat this step three times, finally washing once with acetonitrile. Dissolve the final white solid crude product in an appropriate amount of water and acetonitrile, separate and purify by preparative RP-HPLC, and freeze-dry the target component to obtain a white flocculent solid NV002 (18 mg, yield 58%).

[0136] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.75(s,1H),8.54(s,1H),8.35-8.21(m,1H),8.14(s,1H),7.85(s,1H),7.74-7.68(m,1H),7 .60(d,J=8.4Hz,1H),7.54(s,1H),7.47(d,J=8.0Hz,1H),7.34(d,J=8.3Hz,1H),7.22-7.19(m,2H), 6.81-6.76(m,1H),6.72(d,J=8.3Hz,1H),6.41-6.38(m,1H),6.24(d,J=2.4Hz,1H),5.77(d,J=7.6 Hz,1H),5.64(s,1H),5.28(d,J=7.7Hz,1H),5.26-5.22(m,2H),5.17(s,1H),5.13-5.09(m,1H),4.8 2(s,1H),4.70-4.64(m,1H),4.46(d,J=5.6Hz,1H),4.39(d,J=5.4Hz,1H),4.23(s,1H),4.12(s,1H ),4.09-4.03(m,1H),3.68(d,J=10.9Hz,1H),3.57-3.50(m,2H),3.33-3.25(m,3H),3.24(d,J=6.2H z, 1H), 3.19(s, 1H), 2.69(s, 1H), 2.17–2.08(m, 1H), 1.95–1.88(m, 1H), 1.76–1.67(m, 2H), 1.65–1.53(m, 2H), 1.42–1.36(m, 1H), 1.31(s, 3H), 1.07(d, J = 6.2 Hz, 3H), 0.91(d, J = 6.3 Hz, 3H). High-resolution mass spectrometry (ESI) + C 68 H 81 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1518.5018, measured value is m / z 1518.4994.

[0137] Example 3: Preparation of compound NV003

[0138]

[0139] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [(4-aminobutyl)amino]methane-2-methylpropyl-2-yl ester. The other required raw materials, reagents and preparation steps were the same as in Example 1, and NV003 (38 mg, yield 75%) was obtained.

[0140] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.52-8.48(m,1H),7.86-7.83(m,1H),7.60(d,J=8.5Hz,1H),7.54(s,1H),7.48-7. 45(m,1H),7.33(d,J=8.4Hz,1H),7.23-7.20(m,2H),6.79-6.76(m,1H),6.71(d,J=8.4Hz,1H),6.38(d, J=2.3Hz,1H),6.24(d,J=2.3Hz,1H),5.76(d,J=7.6Hz,1H),5.64(s,1H),5.28(d,J=7.7Hz,1H),5.26-5 .23(m,2H),5.18-5.16(m,1H),5.13-5.10(m,1H),4.83-4.81(m,1H),4.69-4.65(m,1H),4.49-4.45(m, 1H),4.38(d,J=5.6Hz,1H),4.22(s,1H),4.12(s,1H),4.07-4.04(m,1H),3.68(d,J=10.8Hz,1H),3.57 -3.49(m,2H),3.30-3.25(m,2H),3.24-3.20(m,2H),3.18(s,1H),3.13-3.08(m,1H),2.79(t,J=7.0Hz, 2H), 2.68(s, 1H), 2.16–2.08(m, 1H), 1.93–1.88(m, 1H), 1.77–1.66(m, 2H), 1.66–1.55(m, 2H), 1.54–1.47(m, 4H), 1.31(s, 3H), 1.07(d, J = 6.3 Hz, 3H), 0.91(d, J = 6.3 Hz, 3H), 0.90(d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI) + C 69 H 83 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1504.5113, measured value is m / z 1504.5094.

[0141] Example 4: Preparation of compound NV004

[0142]

[0143] In Example 2, NV001 was replaced with NV003, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV004 (22 mg, yield 71%).

[0144] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.49(s,1H),7.99(s,1H),7.85(s,1H),7.59(d,J=8.5Hz,1H),7.54(s,1H),7.4 8-7.45(m,1H),7.33(d,J=8.3Hz,1H),7.23-7.18(m,2H),6.77(d,J=8.4Hz,1H),6.71(d,J=8.4Hz,1 H),6.38(d,J=2.3Hz,1H),6.25(d,J=2.4Hz,1H),5.76(d,J=7.6Hz,1H),5.64(s,1H),5.28(d,J=7. 8Hz,1H),5.27-5.22(m,2H),5.16(s,1H),5.12-5.07(m,1H),4.85-4.78(m,1H),4.70-4.63(m,1H), 4.46(d,J=5.6Hz,1H),4.38(d,J=5.6Hz,1H),4.22(s,1H),4.12(s,1H),4.07-4.03(m,1H),3.68(d ,J=10.6Hz,1H),3.58-3.50(m,3H),3.30-3.23(m,2H),3.23-3.16(m,2H),3.15-3.11(m,1H),3.11- 3.05 (m, 2H), 2.73–2.67 (m, 1H), 2.16–2.08 (m, 1H), 1.95–1.88 (m, 1H), 1.77–1.67 (m, 2H), 1.65–1.53 (m, 2H), 1.51–1.43 (m, 5H), 1.31 (s, 3H), 1.07 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 70 H 85 Cl2N 13 O 23 Theoretical value [M+H] +m / z 1546.5331, measured value is m / z 1546.5301.

[0145] Example 5: Preparation of compound NV005

[0146]

[0147] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [(6-aminohexyl)amino]methane-2-methylpropyl-2-yl ester, and the remaining raw materials, reagents and preparation steps were the same as in Example 1, to obtain NV005 (35 mg, yield 67%).

[0148] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.48(s,1H),7.87-7.83(m,1H),7.60(d,J=8.5Hz,1H),7.54(s,1H),7.49-7.45(m,1H), 7.33(d,J=8.4Hz,1H),7.23-7.19(m,2H),6.82(d,J=11.5Hz,1H),6.77(d,J=8.4Hz,1H),6.71(d,J=8.4Hz, 1H),6.38(d,J=2.3Hz,1H),6.25(d,J=2.4Hz,1H),5.76(d,J=7.5Hz,1H),5.63(s,1H),5.28(d,J=7.8Hz,1H ),5.25-5.20(m,1H),5.18-5.15(m,1H),5.13-5.08(m,1H),4.85-4.77(m,1H),4.67(d,J=6.7Hz,1H),4.50- 4.44(m,1H),4.37(d,J=5.7Hz,1H),4.26-4.20(m,1H),4.12(s,1H),4.08-4.02(m,1H),3.68(d,J=10.7Hz, 1H),3.59-3.50(m,2H),3.32-3.24(m,2H),3.21-3.16(m,2H),3.14-3.09(m,1H),2.77(t,J=7.6Hz,2H),2.1 6-2.09 (m, 1H), 1.91 (d, J = 12.5 Hz, 1H), 1.76-1.66 (m, 2H), 1.64-1.55 (m, 2H), 1.54-1.49 (m, 2H), 1.47-1.42 (m, 2H), 1.34-1.23 (m, 6H), 1.07 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.3 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 71 H 87 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1532.5426, measured value is m / z 1532.5393.

[0149] Example 6: Preparation of compound NV006

[0150]

[0151] In Example 2, NV001 was replaced with NV005, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV006 (22 mg, yield 71%).

[0152] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.73(s,1H),8.46(s,1H),7.87-7.84(m,1H),7.60(d,J=8.4Hz,1H),7.54(s,1H),7.48-7.45 (m,1H),7.32(d,J=8.4Hz,1H),7.23-7.18(m,2H),6.76(d,J=8.4Hz,1H),6.71(d,J=8.4Hz,1H),6.3 8(d,J=2.3Hz,1H),6.25(d,J=2.3Hz,1H),5.76(d,J=7.6Hz,1H),5.64(s,1H),5.27(d,J=7.7Hz,1H) ,5.25-5.22(m,2H),5.16(s,1H),5.13-5.10(m,1H),4.82(d,J=3.5Hz,1H),4.69-4.64(m,1H),4.45 (d,J=5.2Hz,1H),4.38(d,J=5.7Hz,1H),4.21(s,1H),4.12(s,1H),4.07-4.03(m,1H),3.68(d,J=1 0.6Hz,1H),3.59-3.50(m,2H),3.29-3.24(m,2H),3.20-3.17(m,1H),3.12-3.06(m,3H),2.73-2.64 (m, 1H), 2.16–2.08 (m, 1H), 1.94–1.88 (m, 1H), 1.76–1.68 (m, 2H), 1.66–1.54 (m, 2H), 1.50–1.43 (m, 4H), 1.33–1.30 (m, 3H), 1.30–1.26 (m, 4H), 1.07 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 72 H 89 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1574.5644, measured value is m / z 1574.5623.

[0153] Example 7: Preparation of compound NV007

[0154]

[0155] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [(8-aminooctyl)amino]methane-2-methylpropyl-2-yl ester, and the other required raw materials, reagents and preparation steps were the same as in Example 1, to obtain NV007 (39 mg, yield 74%).

[0156] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.73(s,1H),8.46(s,1H),7.88-7.83(m,1H),7.59(d,J=8.4Hz,1H),7.54(s,1H),7.46(d,J=8.3Hz, 1H),7.32(d,J=8.4Hz,1H),7.23-7.17(m,1H),6.76(d,J=8.4Hz,1H),6.71(d,J=8.4Hz,1H),6.37(d,J=2. 3Hz,1H),6.25(d,J=2.4Hz,1H),5.76(d,J=7.6Hz,1H),5.64(s,1H),5.27(d,J=7.7Hz,1H),5.26-5.22(m, 1H),5.16(s,1H),5.13-5.10(m,1H),4.85-4.80(m,1H),4.70-4.64(m,1H),4.45(d,J=5.6Hz,1H),4.38(d ,J=5.7Hz,1H),4.21(s,1H),4.12(s,1H),4.07-4.02(m,1H),3.68(d,J=10.6Hz,1H),3.57-3.51(m,1H),3 .30-3.24(m,1H),3.18(s,1H),3.16-3.07(m,1H),2.77(t,J=7.6Hz,2H),2.69(s,1H),2.17-2.09(m,1H), 1.91 (d, J = 12.4 Hz, 1H), 1.77–1.66 (m, 2H), 1.65–1.55 (m, 2H), 1.55–1.49 (m, 2H), 1.49–1.42 (m, 2H), 1.31 (s, 3H), 1.29–1.21 (m, 6H), 1.07 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.2 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 73 H 91 Cl2N 11 O 23 Theoretical value [M+H]+ m / z 1560.5739, measured value is m / z 1560.5705.

[0157] Example 8: Preparation of compound NV008

[0158]

[0159] In Example 2, NV001 was replaced with NV007, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV008 (24 mg, yield 75%).

[0160] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.73(s,1H),8.45(d,J=5.2Hz,1H),7.92-7.84(m,2H),7.59(d,J=8.5Hz,1H),7.54(s,1H),7.4 7-7.44(m,1H),7.32(d,J=8.4Hz,1H),7.22-7.18(m,2H),6.76(d,J=8.5Hz,1H),6.71(d,J=8.5Hz,1H) ,6.37(d,J=2.3Hz,1H),6.25(d,J=2.4Hz,1H),5.76(d,J=7.6Hz,1H),5.64(s,1H),5.27(d,J=7.7Hz, 1H),5.26-5.22(m,2H),5.16(s,1H),5.12-5.10(m,1H),4.82(d,J=3.6Hz,1H),4.70-4.64(m,1H),4.4 7-4.44(m,1H),4.38(d,J=5.7Hz,1H),4.21(s,1H),4.12(s,1H),4.07-4.04(m,1H),3.68(d,J=10.6H z,1H),3.58-3.50(m,2H),3.30-3.24(m,2H),3.20-3.16(m,1H),3.15-3.11(m,1H),3.10-3.05(m,3H) 2.73–2.65 m, 1H; 2.16–2.09 m, 1H; 1.91 d, J = 9.5 Hz, 1H; 1.77–1.67 m, 2H; 1.67–1.55 m, 2H; 1.51–1.42 m, 5H; 1.31 s, 3H; 1.28 s, 9H; 1.07 d, J = 6.3 Hz, 3H; 0.91 d, J = 6.5 Hz, 3H. High-resolution mass spectrometry (ESI) + C 74 H93 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1602.5957, measured value is m / z 1602.5939.

[0161] Example 9: Preparation of compound NV009

[0162]

[0163] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [(8-amino-3,6-dioxaoct-1-yl)amino]methane-2-methylpropyl-2-yl ester, and the remaining raw materials, reagents and preparation steps were the same as in Example 1, to obtain NV009 (32 mg, yield 60%).

[0164] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.50(s,1H),7.88-7.83(m,1H),7.60(d,J=8.5Hz,1H),7.54(s,1H),7.46(d,J=8.3Hz, 1H),7.33(d,J=8.3Hz,1H),7.22-7.17(m,2H),6.77(d,J=8.4Hz,1H),6.71(d,J=8.4Hz,1H),6.39(d,J=2.4 Hz,1H),6.26(d,J=2.3Hz,1H),5.76(d,J=7.6Hz,1H),5.64(s,1H),5.27(d,J=7.7Hz,1H),5.26-5.21(m,2 H),5.18-5.15(m,1H),5.14-5.09(m,1H),4.83-4.81(m,1H),4.67(q,J=6.6Hz,1H),4.46(d,J=5.5Hz,1H), 4.40(d,J=5.8Hz,1H),4.23-4.20(m,1H),4.12(s,1H),4.08-4.02(m,1H),3.68(d,J=10.7Hz,1H),3.62-3 .56(m,J=2.7Hz,5H),3.56-3.53(m,1H),3.52-3.48(m,2H),3.36-3.29(m,2H),3.29-3.23(m,2H),3.18(s, 1H), 2.96 (t, J = 5.2 Hz, 2H), 2.68 (s, 1H), 2.16–2.09 (m, 1H), 1.91 (d, J = 9.3 Hz, 1H), 1.77–1.67 (m, 2H), 1.65–1.54 (m, 2H), 1.31 (s, 3H), 1.07 (d, J = 6.4 Hz, 3H), 0.91 (d, J = 6.2 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 71 H 87 Cl2N 11 O 25 Theoretical value [M+H] + m / z 1564.5325, measured value is m / z 1564.5301.

[0165] Example 10: Preparation of compound NV010

[0166]

[0167] In Example 2, NV001 was replaced with NV009, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV010 (18 mg, yield 56%).

[0168] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.49(s,1H),7.97(s,1H),7.89-7.84(m,1H),7.59(d,J=8.4Hz,1H),7.54(s,1H),7. 48-7.45(m,1H),7.32(d,J=8.3Hz,1H),7.21(d,J=8.4Hz,1H),7.19(s,1H),6.77(d,J=8.4Hz,1H),6.71( d,J=8.4Hz,1H),6.38(d,J=2.4Hz,1H),6.25(d,J=2.4Hz,1H),5.76(d,J=7.6Hz,1H),5.64(s,1H),5.27 (d,J=7.7Hz,1H),5.26-5.21(m,2H),5.16(d,J=2.0Hz,1H),5.13-5.10(m,1H),4.85-4.78(m,1H),4.71- 4.64(m,1H),4.48-4.44(m,1H),4.40(d,J=5.7Hz,1H),4.21(s,1H),4.12(s,1H),4.08-4.03(m,1H),3. 68(d,J=10.7Hz,1H),3.61-3.56(m,5H),3.55-3.53(m,1H),3.52-3.49(m,2H),3.38-3.29(m,2H),3.28- 3.22 (m, 4H), 3.18 (s, 1H), 2.68 (s, 1H), 2.16–2.09 (m, 1H), 1.94–1.89 (m, 1H), 1.76–1.67 (m, 2H), 1.65–1.53 (m, 2H), 1.31 (s, 3H), 1.07 (d, J = 6.4 Hz, 3H), 0.91 (d, J = 6.4 Hz, 3H), 0.90 (d, J = 6.4 Hz, 3H). High-resolution mass spectrometry (ESI) + C 72 H 89 Cl2N 13 O 25 Theoretical value [M+H] + m / z 1606.5543, measured value is m / z 1606.5523.

[0169] Example 11: Preparation of compound NV011

[0170]

[0171] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with ({[4-(aminomethyl)phenyl]methyl}amino)methane-2-methylpropyl-2-yl ester, and the other required raw materials, reagents and preparation steps were the same as in Example 1, to obtain NV011 (35 mg, yield 66%).

[0172] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.75-8.72(m,1H),8.55-8.49(m,2H),7.85(s,1H),7.60(d,J=8.5Hz,1H),7.53(s,1H),7.46(d,J=8. 3Hz,1H),7.36(d,J=8.3Hz,2H),7.33-7.30(m,3H),7.22-7.18(m,2H),6.78(d,J=1.9Hz,1H),6.71(d,J=8.4 Hz,1H),6.39(d,J=2.4Hz,1H),6.30(d,J=2.3Hz,1H),5.76(d,J=7.6Hz,1H),5.63(s,1H),5.27(d,J=7.8Hz ,1H),5.25-5.21(m,2H),5.16(s,1H),5.11(d,J=3.4Hz,1H),4.81(d,J=3.6Hz,1H),4.66(q,J=6.6Hz,1H),4 .49-4.42(m,3H),4.36-4.30(m,1H),4.24(s,1H),4.13(s,1H),4.05(d,J=8.6Hz,1H),4.00-3.96(m,2H),3 .67(d,J=10.9Hz,1H),3.58-3.52(m,2H),3.46-3.41(m,2H),3.30-3.23(m,2H),3.18(s,1H),2.72-2.64(m, 1H), 2.54(s, 1H), 2.14–2.08(m, 1H), 2.06(s, 1H), 1.93–1.88(m, 1H), 1.73(d, J = 12.9 Hz, 1H), 1.69–1.51(m, 4H), 1.30(s, 3H), 1.23(s, 1H), 1.07–1.02(m, 4H), 0.91(d, J = 6.4 Hz, 3H), 0.90(d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C73 H 83 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1552.5113, measured value is m / z 1552.5098.

[0173] Example 12: Preparation of compound NV012

[0174]

[0175] In Example 2, NV001 was replaced with NV011, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV012 (19 mg, yield 59%).

[0176] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.73(s,1H),8.54-8.51(m,1H),8.48(s,1H),7.86-7.84(m,1H),7.60(d,J=8.5Hz,1H),7.53(s,1H ),7.47-7.44(m,1H),7.32(d,J=8.3Hz,1H),7.29-7.26(m,2H),7.24-7.19(m,3H),6.79-6.75(m,1H),6. 71(d,J=8.4Hz,1H),6.38(d,J=2.3Hz,1H),6.31(d,J=2.3Hz,1H),5.76(d,J=7.4Hz,1H),5.63(s,1H),5. 27(d,J=7.7Hz,1H),5.25-5.23(m,2H),5.16(s,1H),5.11(d,J=3.4Hz,1H),4.81(d,J=3.6Hz,1H),4.69-4 .64(m,1H),4.49-4.45(m,1H),4.43(d,J=4.9Hz,1H),4.35-4.29(m,3H),4.24(s,1H),4.13(s,1H),4.07 -4.03(m,1H),3.67(d,J=10.7Hz,1H),3.57-3.52(m,2H),3.45-3.41(m,3H),3.27-3.24(m,2H),3.18(s,1 2.73–2.64 (m, 1H), 2.54 (s, 1H), 2.15–2.08 (m, 1H), 1.90 (d, J = 9.9 Hz, 1H), 1.73 (d, J = 13.0 Hz, 1H), 1.70–1.53 (m, 4H), 1.30 (s, 3H), 1.08–1.03 (m, 4H), 0.91 (d, J = 6.4 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 74 H 85 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1594.5331, measured value is m / z 1594.5334.

[0177] Example 13: Preparation of compound NV013

[0178]

[0179] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [({4-[4-(aminomethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (compound 1), and the other required raw materials, reagents and preparation steps were the same as in Example 1, to obtain NV013 (36 mg, yield 65%).

[0180] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.59-8.53(m,1H),8.53-8.49(m,1H),7.88-7.84(m,1H),7.78(s,1H),7.70-7.67(m,1H), 7.63(d,J=8.1Hz,2H),7.60(d,J=8.9Hz,1H),7.53(d,J=6.0Hz,1H),7.51(d,J=7.7Hz,1H),7.49-7.45(m,1H), 7.44-7.42(m,1H),7.40(d,J=8.1Hz,2H),7.33(d,J=8.3Hz,1H),7.24-7.19(m,2H),6.78(d,J=8.4Hz,1H),6.7 2(d,J=8.4Hz,1H),6.39(d,J=2.3Hz,1H),6.33(d,J=2.3Hz,1H),5.77(d,J=7.2Hz,1H),5.64(s,1H),5.29-5.2 4(m,2H),5.17(s,1H),5.11(s,1H),4.83-4.80(m,1H),4.70-4.65(m,1H),4.53-4.50(m,1H),4.50-4.45(m,2H ),4.43-4.37(m,1H),4.26(s,1H),4.16-4.09(m,3H),4.07-4.03(m,1H),3.68(d,J=10.8Hz,1H),3.57-3.50(m ,2H), 3.30-3.23 (m,2H), 3.18 (s,1H), 2.69 (s,1H), 2.16-2.09 (m,1H), 1.94-1.88 (m,1H), 1.76-1.66 (m,2H), 1.65-1.53 ​​(m,2H), 1.31 (s,3H), 1.07 (d,J=6.3Hz,3H), 0.91 (d,J=6.5Hz,3H), 0.90 (d,J=6.5Hz,3H). High-resolution mass spectrometry (ESI) + C 79 H 87 Cl2N 11 O23 Theoretical value [M+H] + m / z 1628.5426, measured value 1628.5393.

[0181] Example 14: Preparation of compound NV014

[0182]

[0183] In Example 2, NV001 was replaced with NV013, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV014 (22 mg, yield 66%).

[0184] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.75(s,1H),8.57-8.54(m,1H),8.53-8.47(m,1H),7.86(s,1H),7.60(s,1H),7.60-7.59(m,2H),7.58(s, 1H),7.54(s,1H),7.50-7.48(m,1H),7.48-7.45(m,1H),7.39(d,J=8.0Hz,2H),7.33(d,J=8.4Hz,1H),7.29(d,J= 7.5Hz,1H),7.23-7.20(m,2H),6.78(d,J=8.4Hz,1H),6.72(d,J=8.4Hz,1H),6.39(d,J=2.3Hz,1H),6.33(d,J=2. 3Hz,1H),5.77(d,J=7.4Hz,1H),5.64(s,1H),5.29-5.26(m,2H),5.25-5.23(m,1H),5.18-5.16(m,1H),5.11(d,J =3.4Hz,1H),4.82(d,J=3.6Hz,1H),4.70-4.64(m,1H),4.48(d,J=5.3Hz,2H),4.44-4.42(m,2H),4.41-4.37(m,1 H),4.26(s,1H),4.12(s,1H),4.05(t,J=7.3Hz,1H),3.68(d,J=10.7Hz,1H),3.55(d,J=8.8Hz,1H),3.54-3.50(m 3.30–3.24 (m, 2H), 3.18 (s, 1H), 2.73 (s, 1H), 2.18–2.08 (m, 1H), 1.91 (d, J = 10.6 Hz, 1H), 1.76–1.66 (m, 2H), 1.65–1.51 (m, 2H), 1.31 (s, 3H), 1.07 (d, J = 6.3 Hz, 3H), 0.92 (d, J = 6.5 Hz, 3H), 0.91 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 80 H 89 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1670.5644, measured value is m / z 1670.5604.

[0185] Example 15: Preparation of compound NV015

[0186]

[0187] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [({4-[4-(2-aminoethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (compound 2). The other required raw materials, reagents and preparation steps were the same as in Example 1, and NV015 (36 mg, yield 64%) was obtained.

[0188] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.75(s,1H),8.50(s,1H),8.13(s,1H),7.90-7.85(m,1H),7.72(d,J=8.1Hz,2H),7.63(d,J=8.1Hz,2H),7. 59(d,J=8.5Hz,1H),7.55-7.51(m,3H),7.49-7.45(m,1H),7.37(d,J=8.1Hz,2H),7.33(d,J=8.3Hz,1H),7.23-7. 20(m,2H),6.79-6.76(m,1H),6.72(d,J=8.5Hz,1H),6.38(d,J=2.3Hz,1H),6.31(d,J=2.3Hz,1H),5.77(d,J=7.4 Hz,1H),5.64(s,1H),5.28(d,J=7.8Hz,1H),5.27-5.23(m,2H),5.19-5.15(m,1H),5.12-5.10(m,1H),4.83-4.80( m,1H),4.70-4.65(m,1H),4.46(s,1H),4.42(d,J=5.6Hz,1H),4.23(s,1H),4.12(s,1H),4.09-4.06(m,2H),4.06 -4.03(m,1H),3.68(d,J=10.8Hz,1H),3.56(d,J=8.8Hz,1H),3.54-3.50(m,2H),3.40-3.36(m,1H),3.30-3.23(m 2H), 3.18 (s, 1H), 2.89–2.80 (m, 2H), 2.74–2.65 (m, 1H), 2.16–2.09 (m, 1H), 1.95–1.87 (m, 1H), 1.77–1.66 (m, 2H), 1.65–1.52 (m, 2H), 1.31 (s, 3H), 1.07 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.6 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C80 H 89 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1642.5583, measured value is m / z 1642.5545.

[0189] Example 16: Preparation of compound NV016

[0190]

[0191] In Example 2, NV001 was replaced with NV015, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV016 (25 mg, yield 74%).

[0192] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.50(s,1H),8.13(d,J=7.9Hz,1H),7.87(s,1H),7.68(d,J=8.2Hz,2H),7.61(d,J=8 .1Hz,2H),7.56-7.52(m,1H),7.49-7.45(m,1H),7.38(d,J=8.2Hz,2H),7.36(d,J=8.3Hz,2H),7.33(d, J=8.3Hz,1H),7.21(d,J=8.4Hz,2H),6.79-6.76(m,1H),6.72(d,J=8.5Hz,1H),6.38(d,J=2.3Hz,1H),6 .31(d,J=2.3Hz,1H),5.76(d,J=7.1Hz,1H),5.64(s,1H),5.29-5.24(m,3H),5.17(s,1H),5.11(d,J=3. 2Hz,1H),4.82(d,J=3.6Hz,1H),4.70-4.65(m,1H),4.46(s,1H),4.43-4.39(m,2H),4.23(s,1H),4.12( s,1H),4.05(t,J=7.4Hz,1H),3.68(d,J=10.4Hz,1H),3.58-3.50(m,2H),3.41-3.36(m,2H),3.31-3.24 (m, 2H), 3.18(s, 1H), 2.87–2.80(m, 2H), 2.75–2.64(m, 2H), 2.15–2.08(m, 1H), 1.94–1.87(m, 2H), 1.76–1.67(m, 2H), 1.64–1.53(m, 2H), 1.31(s, 3H), 1.07(d, J = 6.3 Hz, 3H), 0.91(d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 91 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1684.5801, measured value is m / z 1684.5754.

[0193] Example 17: Preparation of compound NV017

[0194]

[0195] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [(2-{4-[4-(aminomethyl)phenyl]phenyl}ethyl)amino]methane-2-methylpropyl-2-yl ester (compound 3). The other required raw materials, reagents and preparation steps were the same as in Example 1, and NV017 (36 mg, yield 65%) was obtained.

[0196] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.61(s,1H),8.44-8.40(m,1H),8.38-8.34(m,1H),7.73-7.72(m,1H),7.50-7.48(m,2H),7.46-7.43(m ,2H),7.42-7.39(m,1H),7.34-7.31(m,1H),7.24(s,1H),7.23-7.22(m,2H),7.21(s,1H),7.19(d,J=8.4Hz,1H ),7.10-7.06(m,2H),6.66-6.63(m,1H),6.58(d,J=8.5Hz,1H),6.26(d,J=2.3Hz,1H),6.19(d,J=2.4Hz,1H), 5.65-5.61(m,1H),5.50(s,1H),5.16-5.12(m,2H),5.12-5.10(m,1H),5.05-5.03(m,1H),4.99-4.96(m,1H),4 .68(d,J=3.6Hz,1H),4.56-4.51(m,1H),4.38-4.35(m,1H),4.35-4.31(m,2H),4.28-4.22(m,1H),4.12(s,1H ),3.99(s,1H),3.94-3.90(m,1H),3.54(d,J=10.9Hz,1H),3.45-3.37(m,3H),3.16-3.10(m,2H),3.04(s,1H), 2.97–2.92 (m, 2H), 2.79–2.74 (m, 2H), 2.56 (s, 1H), 2.04–1.95 (m, 1H), 1.80–1.74 (m, 1H), 1.64–1.52 (m, 2H), 1.52–1.40 (m, 2H), 1.17 (s, 3H), 0.93 (d, J = 6.3 Hz, 3H), 0.78 (d, J = 6.4 Hz, 3H), 0.77 (d, J = 6.4 Hz, 3H). High-resolution mass spectrometry (ESI) + C 80 H 89 Cl2N 11 O23 Theoretical value [M+H] + m / z 1642.5583, measured value is m / z 1642.5557.

[0197] Example 18: Preparation of compound NV018

[0198]

[0199] In Example 2, NV001 was replaced with NV017, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV018 (26 mg, yield 77%).

[0200] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.61(s,1H),8.43-8.39(m,1H),8.38-8.35(m,1H),7.73(s,1H),7.49-7.46(m,3H),7.45(d,J=8.1H z,2H),7.41(s,1H),7.35-7.32(m,1H),7.26-7.22(m,3H),7.22(s,1H),7.20(d,J=8.4Hz,1H),7.11-7.04 (m,2H),6.65(d,J=8.4Hz,1H),6.59(d,J=8.4Hz,1H),6.26(d,J=2.3Hz,1H),6.19(d,J=2.4Hz,1H),5.63( d,J=7.1Hz,1H),5.51(s,1H),5.16-5.13(m,2H),5.12(d,J=5.1Hz,1H),5.04(s,1H),4.98(d,J=3.4Hz,1H) ,4.69(d,J=3.6Hz,1H),4.56-4.52(m,1H),4.38-4.32(m,3H),4.28-4.22(m,1H),4.12(s,1H),4.00(s,1H ),3.94-3.90(m,1H),3.55(d,J=10.6Hz,1H),3.42(d,J=8.8Hz,1H),3.41-3.36(m,2H),3.29-3.25(m,3H) 3.17–3.10 (m, 2H), 3.05 (s, 1H), 2.72–2.68 (m, 2H), 2.56 (s, 1H), 2.03–1.95 (m, 1H), 1.80–1.74 (m, 1H), 1.62–1.54 (m, 2H), 1.52–1.40 (m, 2H), 1.18 (s, 3H), 0.94 (d, J = 6.3 Hz, 3H), 0.78 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 91 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1684.5801, measured value is m / z 1684.5781.

[0201] Example 19: Preparation of compound NV019

[0202]

[0203] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [({3-[4-(aminomethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (compound 4). The other required raw materials, reagents and preparation steps were the same as in Example 1, and NV019 (36 mg, yield 65%) was obtained.

[0204] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.59(s,1H),8.41-8.35(m,2H),7.72-7.69(m,1H),7.62(s,1H),7.54-7.50(m,1H),7.47(d,J=8.0Hz,2H),7. 44(d,J=8.7Hz,1H),7.38(d,J=6.7Hz,1H),7.35(d,J=7.7Hz,1H),7.32-7.30(m,1H),7.28-7.26(m,1H),7.24(d,J=8 .1Hz,2H),7.17(d,J=8.4Hz,1H),7.08-7.03(m,2H),6.63(d,J=8.4Hz,1H),6.56(d,J=8.3Hz,1H),6.24(d,J=2.3Hz, 1H),6.17(d,J=2.3Hz,1H),5.61(d,J=7.7Hz,1H),5.48(s,1H),5.13-5.10(m,2H),5.09(d,J=4.0Hz,1H),5.02-5.00 (m,1H),4.96(d,J=3.1Hz,1H),4.66(d,J=3.5Hz,1H),4.51(q,J=6.6Hz,1H),4.34-4.30(m,2H),4.27-4.21(m,1H), 4.10(s,1H),3.96-3.94(m,2H),3.91-3.87(m,1H),3.52(d,J=10.9Hz,1H),3.41-3.37(m,2H),3.30-3.26(m,2H),3. 13-3.10 (m, 2H), 3.02 (s, 1H), 2.57-2.50 (m, 1H), 2.39 (s, 1H), 1.99-1.92 (m, 1H), 1.77-1.72 (m, 1H), 1.57 (d, J = 13.1 Hz, 1H), 1.54-1.38 (m, 4H), 1.15 (s, 3H), 0.94-0.88 (m, 4H), 0.75 (d, J = 6.5 Hz, 3H), 0.75 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 79 H87 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1628.5426, measured value is m / z 1628.5402.

[0205] Example 20: Preparation of compound NV020

[0206]

[0207] In Example 2, NV001 was replaced with NV019, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV020 (21 mg, yield 63%).

[0208] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.57-8.50(m,2H),7.87-7.84(m,1H),7.61-7.56(m,4H),7.55-7.52(m,1H),7.50-7.44(m ,2H),7.39(d,J=8.0Hz,2H),7.32(d,J=8.4Hz,1H),7.29(d,J=7.7Hz,1H),7.23-7.19(m,2H),6.78(d,J=8.4Hz ,1H),6.72(d,J=8.4Hz,1H),6.39(d,J=2.3Hz,1H),6.32(d,J=2.3Hz,1H),5.78-5.73(m,1H),5.63(s,1H),5. 29-5.26(m,2H),5.24(d,J=4.0Hz,1H),5.17-5.15(m,1H),5.13-5.10(m,1H),4.81(d,J=3.6Hz,1H),4.69-4.6 4(m,1H),4.49-4.46(m,2H),4.44-4.42(m,2H),4.39(d,J=15.4Hz,1H),4.25(s,1H),4.14-4.09(m,1H),4.07 -4.03(m,1H),3.67(d,J=10.8Hz,1H),3.57-3.52(m,2H),3.45-3.41(m,2H),3.29-3.24(m,2H),3.18(s,1H),2 0.72–2.65 (m, 1H), 2.54 (s, 1H), 2.15–2.07 (m, 1H), 1.93–1.87 (m, 1H), 1.73 (d, J = 13.1 Hz, 1H), 1.70–1.54 (m, 4H), 1.30 (s, 3H), 1.24–1.21 (m, 1H), 1.09–1.03 (m, 4H), 0.91 (d, J = 6.5 Hz, 3H), 0.90 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C 80 H 89 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1670.5644, measured value is m / z 1670.5629.

[0209] Example 21: Preparation of compound NV021

[0210]

[0211] The [(2-aminoethyl)amino]methane-2-methylpropyl-2-yl ester in Example 1 was replaced with [({2-[4-(aminomethyl)phenyl]phenyl}methyl)amino]methane-2-methylpropyl-2-yl ester (compound 5). The other required raw materials, reagents and preparation steps were the same as in Example 1, and NV021 (39 mg, yield 70%) was obtained.

[0212] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.59(s,1H),8.44-8.36(m,2H),7.70(s,1H),7.44(d,J=7.7Hz,2H),7.38(s,1H),7.35-7.29(m ,3H),7.24(d,J=7.9Hz,2H),7.19-7.12(m,4H),7.08-7.02(m,2H),6.62(d,J=8.4Hz,1H),6.57(d,J=8 .4Hz,1H),6.24(d,J=2.3Hz,1H),6.17(d,J=2.3Hz,1H),5.61(d,J=7.5Hz,1H),5.48(s,1H),5.14-5. 08(m,3H),5.01(s,1H),4.96(d,J=3.4Hz,1H),4.66(d,J=3.6Hz,1H),4.51(q,J=6.6Hz,1H),4.40-4.3 5(m,1H),4.33-4.31(m,2H),4.28-4.23(m,1H),4.10(s,1H),3.98(s,1H),3.92-3.87(m,1H),3.84-3 .79(m,2H),3.52(d,J=10.9Hz,1H),3.42-3.37(m,2H),3.30-3.25(m,2H),3.14-3.08(m,2H),3.03(s, 1H), 2.58-2.47 (m, 1H), 2.39 (s, 1H), 2.01-1.93 (m, 1H), 1.78-1.73 (m, 1H), 1.60-1.56 (m, 1H), 1.54-1.37 (m, 4H), 1.15 (s, 3H), 0.93-0.88 (m, 4H), 0.75 (d, J = 6.2 Hz, 3H), 0.75 (d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI) + C 79 H 87 Cl2N 11 O 23 Theoretical value [M+H] +m / z 1628.5426, measured value is m / z 1628.5434.

[0213] Example 22: Preparation of compound NV022

[0214]

[0215] In Example 2, NV001 was replaced with NV021, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV022 (22 mg, yield 66%).

[0216] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.59(s,1H),8.42-8.36(m,2H),7.71-7.69(m,1H),7.61-7.55(m,1H),7.44(d,J=8.4Hz,1H),7.3 9(s,1H),7.32-7.25(m,3H),7.24-7.20(m,3H),7.17(d,J=8.4Hz,1H),7.14-7.10(m,2H),7.08-7.04(m, 2H),6.65-6.61(m,1H),6.57(d,J=8.4Hz,1H),6.24(d,J=2.3Hz,1H),6.17(d,J=2.3Hz,1H),5.61(d,J= 7.6Hz,1H),5.48(s,1H),5.14-5.07(m,3H),5.01(s,1H),4.96(d,J=3.3Hz,1H),4.66(d,J=3.5Hz,1H),4 .53-4.49(m,1H),4.39-4.34(m,1H),4.34-4.31(m,2H),4.28-4.23(m,1H),4.11-4.06(m,3H),3.97(s, 1H),3.92-3.87(m,1H),3.52(d,J=10.8Hz,1H),3.42-3.37(m,2H),3.30-3.26(m,2H),3.14-3.08(m,2H) 3.02 (s, 1H), 2.57–2.50 (m, 1H), 2.00–1.93 (m, 1H), 1.75 (d, J = 12.5 Hz, 1H), 1.57 (d, J = 13.3 Hz, 1H), 1.55–1.38 (m, 4H), 1.15 (s, 3H), 0.93–0.88 (m, 4H), 0.76 (d, J = 6.5 Hz, 3H), 0.75 (d, J = 6.5 Hz, 3H). High-resolution mass spectrometry (ESI) + C80 H 89 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1670.5644, measured value is m / z 1670.5635.

[0217] Example 23: Preparation of compound NV023

[0218]

[0219] In Example 13, vancomycin hydrochloride was replaced with vancomycin hydrochloride. The other required raw materials, reagents and preparation methods were the same as in Example 1, and NV023 (39 mg, yield 70%) was obtained.

[0220] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ7.78(s,1H),7.68(d,J=8.0Hz,1H),7.62(d,J=8.1Hz,2H),7.58-7.55(m,1H),7.52(d,J=7.7Hz,2H),7.5 0(s,1H),7.48-7.46(m,1H),7.44-7.42(m,1H),7.40(d,J=8.2Hz,2H),7.33(d,J=8.3Hz,1H),7.25-7.22(m,1H ),7.20(d,J=8.4Hz,1H),6.78(d,J=8.3Hz,1H),6.73(d,J=8.4Hz,1H),6.40(d,J=2.3Hz,1H),6.32(d,J=2.3Hz ,1H),5.76(d,J=7.8Hz,1H),5.60(s,1H),5.29-5.26(m,2H),5.24(d,J=4.1Hz,1H),5.20-5.18(m,2H),4.93(s ,1H),4.68(q,J=6.6Hz,1H),4.51(d,J=5.8Hz,1H),4.50-4.45(m,2H),4.42-4.36(m,1H),4.30-4.22(m,1H),4 .13-4.08(m,2H),3.97(s,1H),3.68(d,J=10.8Hz,1H),3.58-3.55(m,1H),3.54-3.53(m,1H),3.45-3.42(m,1H ), 3.28-3.25 (m, 2H), 3.18 (s, 1H), 2.64 (s, 3H), 2.18-2.10 (m, 1H), 1.93-1.88 (m, 1H), 1.74 (d, J = 13.0 Hz, 1H), 1.71-1.53 ​​(m, 4H), 1.30 (s, 3H), 1.07 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.2 Hz, 3H), 0.86 (d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI) + C 80 H 89 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1642.5583, measured value is m / z 1642.5575.

[0221] Example 24: Preparation of compound NV024

[0222]

[0223] In Example 2, NV001 was replaced with NV023, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV024 (22 mg, yield 65%).

[0224] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.56(s,1H),8.36(s,1H),7.87-7.83(m,1H),7.70(s,1H),7.47-7.42(m,4H),7.41-7.37(m,2H), 7.35-7.30(m,2H),7.23(d,J=8.1Hz,2H),7.17(d,J=8.3Hz,1H),7.13(d,J=7.4Hz,1H),7.08-7.04(m,2 H),6.64-6.62(m,1H),6.57(d,J=8.5Hz,1H),6.24(d,J=2.3Hz,1H),6.17(d,J=2.3Hz,1H),5.60(d,J=7 .7Hz,1H),5.45(s,1H),5.13-5.10(m,2H),5.09-5.07(m,1H),5.04-5.02(m,2H),4.76(s,1H),4.55-4. 49(m,1H),4.37-4.34(m,1H),4.33-4.30(m,2H),4.29-4.26(m,2H),4.25-4.21(m,1H),4.10(s,1H),4. 05(s,1H),3.81(s,1H),3.52(d,J=10.8Hz,1H),3.42-3.39(m,2H),3.29-3.26(m,1H),3.12-3.09(m,2H ), 3.02(s, 1H), 2.47(s, 3H), 2.03-1.95(m, 1H), 1.75(d, J = 11.6 Hz, 1H), 1.58(d, J = 13.0 Hz, 1H), 1.55-1.38(m, 4H), 1.15(s, 3H), 0.91(d, J = 6.4 Hz, 3H), 0.76(d, J = 6.1 Hz, 3H), 0.71(d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 91 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1684.5801, measured value is m / z 1684.5833.

[0225] Example 25: Preparation of compound NV025

[0226]

[0227] In Example 15, vancomycin hydrochloride was replaced with vancomycin hydrochloride. The other raw materials, reagents and preparation steps were the same as in Example 1, and NV025 (38 mg, yield 67%) was obtained.

[0228] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.71(s,1H),8.47(s,1H),8.13(d,J=6.6Hz,1H),7.88-7.86(m,1H),7.72(d,J=8.2Hz,2H),7.63(d,J=8.0H z,2H),7.56(d,J=7.8Hz,1H),7.53(d,J=8.3Hz,2H),7.48(d,J=8.3Hz,1H),7.37(d,J=8.1Hz,2H),7.33(d,J=8.3 Hz,1H),7.20(d,J=8.4Hz,2H),6.77(d,J=8.4Hz,1H),6.72(d,J=8.5Hz,1H),6.39(d,J=2.3Hz,1H),6.30(d,J=2. 4Hz,1H),5.77-5.74(m,1H),5.60(s,1H),5.29-5.26(m,2H),5.24(d,J=4.0Hz,1H),5.20-5.17(m,1H),4.92(s,1 H),4.71-4.65(m,1H),4.46(d,J=5.4Hz,1H),4.41(d,J=5.6Hz,1H),4.23(s,1H),4.07(s,2H),3.96(s,1H),3.68 (d,J=10.7Hz,1H),3.57-3.55(m,1H),3.55-3.52(m,1H),3.45-3.43(m,1H),3.39(q,J=7.3Hz,2H),3.29-3.26(m 2H), 3.19(s, 1H), 2.88–2.79(m, 2H), 2.63(s, 3H), 2.19–2.12(m, 1H), 1.94–1.88(m, 1H), 1.74(d, J = 13.0 Hz, 1H), 1.71–1.53(m, 4H), 1.31(s, 3H), 1.07(d, J = 6.4 Hz, 3H), 0.91(d, J = 6.2 Hz, 3H), 0.87(d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI) +C 81 H 91 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1656.5739, measured value is m / z 1656.5730.

[0229] Example 26: Preparation of compound NV026

[0230]

[0231] In Example 2, NV001 was replaced with NV025, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV026 (23 mg, yield 68%).

[0232] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.71(s,1H),8.47(s,1H),8.12(s,1H),7.88-7.84(m,1H),7.67(d,J=8.1Hz,2H),7.61(d,J=7.9Hz,2 H),7.55(d,J=8.6Hz,1H),7.49-7.46(m,1H),7.38(d,J=8.2Hz,2H),7.35(d,J=8.2Hz,2H),7.33(d,J=8.4H z,1H),7.22-7.19(m,2H),6.78-6.76(m,1H),6.72(d,J=8.5Hz,1H),6.38(d,J=2.3Hz,1H),6.30(d,J=2.3H z,1H),5.76(d,J=7.4Hz,1H),5.60(s,1H),5.29-5.23(m,3H),5.19-5.15(m,2H),4.92(s,1H),4.71-4.65(m ,1H),4.45(d,J=5.4Hz,1H),4.42-4.39(m,3H),4.23(s,1H),3.96(s,1H),3.68(d,J=10.8Hz,1H),3.59-3. 53(m,2H),3.47-3.42(m,2H),3.40-3.35(m,2H),3.29-3.24(m,2H),3.18(s,1H),2.87-2.78(m,2H),2.63( 2.54(s, 1H), 2.18–2.12(m, 1H), 1.91(d, J = 12.2 Hz, 1H), 1.73(d, J = 13.0 Hz, 1H), 1.71–1.53(m, 4H), 1.31(s, 3H), 1.08–1.06(m, 3H), 1.06–1.04(m, 2H), 0.91(d, J = 6.2 Hz, 3H), 0.86(d, J = 6.3 Hz, 3H). High-resolution mass spectrometry (ESI) + C 82 H 93 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1698.5957, measured value is m / z 1698.5962.

[0233] Example 27: Preparation of compound NV027

[0234]

[0235] In Example 17, vancomycin hydrochloride was replaced with vancomycin hydrochloride. The other required raw materials, reagents and preparation methods were the same as in Example 1, and NV027 (38 mg, yield 67%) was obtained.

[0236] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.71(s,1H),8.54-8.49(m,2H),7.86(s,1H),7.62(d,J=8.1Hz,2H),7.59-7.56(m,2H),7.55(s ,1H),7.47(d,J=8.3Hz,1H),7.39-7.36(m,3H),7.35-7.31(m,2H),7.22(s,1H),7.20(d,J=8.4Hz,1H) ,6.79(d,J=1.9Hz,1H),6.72(d,J=8.5Hz,1H),6.39(d,J=2.3Hz,1H),6.32(d,J=2.3Hz,1H),5.76(d,J =7.8Hz,1H),5.60(s,1H),5.28-5.26(m,2H),5.24(d,J=4.0Hz,1H),5.20-5.16(m,2H),4.92(s,1H),4 .70-4.65(m,1H),4.49-4.45(m,2H),4.41-4.36(m,1H),4.29-4.16(m,2H),3.97(s,1H),3.68(d,J=1 0.8Hz,1H),3.58-3.53(m,2H),3.28-3.26(m,2H),3.18(s,1H),3.11-3.05(m,2H),2.91-2.87(m,2H), 2.63 (s, 3H), 2.17–2.11 (m, 1H), 1.91 (d, J = 9.5 Hz, 1H), 1.73 (d, J = 13.0 Hz, 1H), 1.70–1.54 (m, 4H), 1.30 (s, 3H), 1.16 (s, 1H), 1.07 (d, J = 6.4 Hz, 3H), 0.91 (d, J = 6.2 Hz, 3H), 0.87 (d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 91 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1656.5739, measured value is m / z 1656.5757.

[0237] Example 28: Preparation of compound NV028

[0238]

[0239] In Example 2, NV001 was replaced with NV027, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV028 (26 mg, yield 74%).

[0240] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.54(s,1H),8.38-8.34(m,1H),7.69(s,1H),7.44(d,J=8.0Hz,2H),7.41(d,J=8.1Hz,2H),7.39 -7.37(m,1H),7.32-7.30(m,1H),7.23-7.18(m,4H),7.17(d,J=8.5Hz,1H),7.06(s,1H),7.04(d,J=8.4 Hz,1H),6.63-6.61(m,1H),6.56(d,J=8.4Hz,1H),6.23(d,J=2.3Hz,1H),6.16(d,J=2.3Hz,1H),5.60( d,J=7.5Hz,1H),5.44(s,1H),5.12-5.09(m,2H),5.08(d,J=4.0Hz,1H),5.04-5.02(m,1H),4.76(s,1H) ,4.51(q,J=6.7Hz,1H),4.32-4.29(m,2H),4.25-4.19(m,1H),4.09(s,1H),3.81(s,1H),3.51(d,J=10 .9Hz,1H),3.41-3.36(m,2H),3.30-3.26(m,2H),3.26-3.22(m,2H),3.13-3.09(m,2H),3.02(s,1H),2. 69-2.65(m, 2H), 2.47(s, 3H), 2.02-1.95(m, 1H), 1.75(d, J = 10.5Hz, 1H), 1.57(d, J = 13.1Hz, 1H), 1.55-1.38(m, 4H), 1.14(s, 3H), 0.92-0.88(m, 4H), 0.75(d, J = 6.1Hz, 3H), 0.70(d, J = 6.2Hz, 3H). High-resolution mass spectrometry (ESI) + C 82 H 93 Cl2N 13 O 23 Theoretical value [M+H] +m / z 1698.5957, measured value is m / z 1698.5967.

[0241] Example 29: Preparation of compound NV029

[0242]

[0243] In Example 7, vancomycin hydrochloride was replaced with vancomycin hydrochloride. The other raw materials, reagents and preparation steps were the same as in Example 1, and NV029 (35 mg, yield 65%) was obtained.

[0244] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.69(s,1H),8.43(s,1H),7.86-7.83(m,1H),7.57-7.51(m,2H),7.47-7.45(m,1H),7.32(d,J=8. 4Hz,1H),7.21-7.18(m,2H),6.76(d,J=8.4Hz,1H),6.71(d,J=8.4Hz,1H),6.37(d,J=2.3Hz,1H),6.24( d,J=2.3Hz,1H),5.75(d,J=7.7Hz,1H),5.60(s,1H),5.26(d,J=7.8Hz,1H),5.24-5.22(m,2H),5.20-5. 17(m,2H),4.92(s,1H),4.70-4.65(m,1H),4.44(d,J=5.5Hz,1H),4.37(d,J=5.7Hz,1H),4.23-4.19(m, 2H),3.98-3.95(m,1H),3.67(d,J=10.9Hz,1H),3.57-3.52(m,2H),3.46-3.40(m,3H),3.28-3.25(m,2H ),3.18(s,1H),3.15-3.08(m,2H),2.78-2.74(m,2H),2.63(s,3H),2.17-2.11(m,1H),1.90(d,J=12.3H z, 1H), 1.73 (d, J = 13.2 Hz, 1H), 1.69–1.54 (m, 4H), 1.54–1.48 (m, 2H), 1.47–1.42 (m, 2H), 1.31 (d, J = 5.1 Hz, 3H), 1.29–1.25 (m, 8H), 1.08–1.03 (m, 5H), 0.91 (d, J = 6.3 Hz, 3H), 0.86 (d, J = 6.3 Hz, 3H). High-resolution mass spectrometry (ESI) + C74 H 93 Cl2N 11 O 23 Theoretical value [M+H] + m / z 1574.5896, measured value is m / z 1574.5891.

[0245] Example 30: Preparation of compound NV030

[0246]

[0247] In Example 2, NV001 was replaced with NV029, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV030 (22 mg, yield 68%).

[0248] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.70(s,1H),8.43(s,1H),7.90(d,J=6.0Hz,1H),7.85(s,1H),7.56-7.52(m,2H),7.48-7. 44(m,1H),7.32(d,J=8.4Hz,1H),7.22-7.18(m,2H),6.76(d,J=8.6Hz,1H),6.71(d,J=8.4Hz,1H) ,6.37(d,J=2.4Hz,1H),6.24(d,J=2.3Hz,1H),5.75(s,1H),5.60(s,1H),5.26(d,J=7.7Hz,1H),5 .24-5.21(m,2H),5.20-5.16(m,2H),4.91(s,1H),4.70-4.64(m,1H),4.44(d,J=5.4Hz,1H),4.37 (d,J=5.6Hz,1H),4.20(s,2H),3.96(s,1H),3.67(d,J=10.8Hz,1H),3.58-3.52(m,2H),3.45-3. 39(m,3H),3.29-3.24(m,2H),3.17(s,1H),3.15-3.10(m,2H),3.10-3.05(m,2H),2.62(s,3H),2. 18–2.11 (m, 1H), 1.90 (d, J = 10.0 Hz, 1H), 1.77–1.72 (m, 1H), 1.68–1.52 (m, 4H), 1.49–1.41 (m, 4H), 1.34–1.19 (m, 11H), 1.10–1.03 (m, 5H), 0.91 (d, J = 6.1 Hz, 3H), 0.86 (d, J = 6.1 Hz, 3H). High-resolution mass spectrometry (ESI) + C 75 H 95 Cl2N 13 O 23 Theoretical value [M+H] + m / z 1616.6114, the measured value is m / z1616.6114.

[0249] Example 31: Preparation of compound NV031

[0250]

[0251] Weigh commercially available vancomycin hydrochloride (50 mg, 0.034 mmol) and [(8-aminooctyl)amino]methane-2-methylpropyl-2-yl ester (66 mg, 0.272 mmol) into a single-necked reaction flask. Add 1 mL of H₂O, 1 mL of CH₃CN, and 120 μL of DIPEA (0.68 mmol) sequentially. Stir vigorously until the reaction solution becomes clear. Transfer the reaction system to -10°C, then add 2.8 μL of 37% formaldehyde aqueous solution (36.72 μmol). Maintain the temperature at -10°C and continue stirring until the reaction stops as detected by analytical reversed-phase high-performance liquid chromatography (RP-HPLC). Add TFA to adjust the reaction solution to a weakly acidic state to quench the reaction. Then add a large amount of acetonitrile to precipitate a white precipitate, which is obtained by centrifugation as a white crude solid. The above operation was repeated twice, followed by deprotection in DCM with 20% TFA at 0°C for 30 min. The target component was separated and purified by preparative RP-HPLC with the addition of appropriate amounts of water and acetonitrile. The target component was freeze-dried to obtain a white flocculent solid NV031 (37 mg, yield 69%).

[0252] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.84-8.80(m,1H),8.67-8.62(m,1H),7.82(s,1H),7.58(s,1H),7.52(d,J=8.5Hz,1H) ,7.47(d,J=8.3Hz,1H),7.32(d,J=8.4Hz,1H),7.22(d,J=8.4Hz,1H),7.11(s,1H),6.86(d,J =8.5Hz,1H),6.79(d,J=8.5Hz,1H),6.54(s,1H),5.75-5.69(m,2H),5.30-5.22(m,2H),5.16 -5.09(m,2H),5.07(s,1H),4.77-4.74(m,1H),4.67(q,J=6.6Hz,1H),4.46(d,J=5.7Hz,1H),4 .43(d,J=5.9Hz,1H),4.14-4.00(m,4H),3.67(d,J=10.9Hz,1H),3.57-3.52(m,2H),3.45-3. 40(m,3H),3.29-3.23(m,2H),3.17(s,1H),2.93-2.86(m,2H),2.79-2.72(m,3H),2.54(s,1H) ,2.16-2.09 (m, 1H), 1.96-1.86 (m, 1H), 1.76-1.68 (m, 2H), 1.67-1.45 (m, 8H), 1.30 (s, 3H), 1.29-1.23 (m, 8H), 1.07-1.03 (m, 5H), 0.91 (d, J = 8.9 Hz, 3H), 0.90 (d, J = 8.9 Hz, 3H). High-resolution mass spectrometry (ESI) + C 74 H 93 Cl2N 11 O 24 Theoretical value [M+H] + m / z 1590.5845, measured value is m / z 1590.5837.

[0253] Example 32: Preparation of compound NV032

[0254]

[0255] In Example 2, NV001 was replaced with NV031, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV032 (26 mg, yield 80%).

[0256] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.84-8.80(m,1H),8.65(s,1H),7.94(s,1H),7.82(s,1H),7.58(s,1H),7.52(d,J=8.6H z,1H),7.47(d,J=8.3Hz,1H),7.32(d,J=8.3Hz,1H),7.22(d,J=8.4Hz,1H),7.11(s,1H),6.86 (d,J=8.4Hz,1H),6.79(d,J=8.5Hz,1H),6.54(s,1H),5.76-5.68(m,2H),5.29-5.23(m,2H),5 .14(s,1H),5.11-5.06(m,2H),4.78-4.74(m,1H),4.67(q,J=6.6Hz,1H),4.46(d,J=5.5Hz,1H ),4.44-4.40(m,1H),4.13-4.00(m,5H),3.67(d,J=10.8Hz,1H),3.57-3.52(m,2H),3.43(q,J =7.1Hz,2H),3.29-3.24(m,2H),3.17(s,1H),3.10-3.04(m,2H),2.89(s,3H),2.80-2.71(m,2 2.18–2.08 (m, 1H), 1.95–1.86 (m, 1H), 1.76–1.68 (m, 2H), 1.68–1.41 (m, 8H), 1.30 (s, 3H), 1.29–1.25 (m, 8H), 1.07–1.03 (m, 4H), 0.91 (d, J = 9.0 Hz, 3H), 0.90 (d, J = 9.0 Hz, 3H). High-resolution mass spectrometry (ESI) + C 75 H 95 Cl2N 13 O 24 Theoretical value [M+H] + m / z 1632.6063, measured value is m / z 1632.6056.

[0257] Example 33: Preparation of compound NV033

[0258]

[0259] In Example 31, vancomycin hydrochloride was replaced with vancomycin hydrochloride. The other raw materials, reagents and preparation steps were the same as in Example 31, and NV033 (39 mg, yield 71%) was obtained.

[0260] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.81-8.77(m,1H),8.64(s,1H),7.81(s,1H),7.62(s,1H),7.53-7.49(m,1H),7.47(d,J=8.4 Hz,1H),7.32(d,J=8.4Hz,1H),7.21(d,J=8.4Hz,1H),7.11(s,1H),6.86(d,J=8.5Hz,1H),6.79(d, J=8.5Hz,1H),6.55(s,1H),5.76-5.67(m,2H),5.29-5.23(m,2H),5.16-5.10(m,3H),4.82(s,1H), 4.67(q,J=6.6Hz,1H),4.46(d,J=5.7Hz,1H),4.43(d,J=5.9Hz,1H),4.14-4.06(m,3H),4.02(d,J= 7.9Hz,1H),3.67(d,J=10.9Hz,1H),3.57-3.53(m,2H),3.45-3.39(m,2H),3.26(d,J=6.0Hz,2H),3 .18(s,1H),2.92-2.86(m,2H),2.80-2.72(m,3H),2.59(s,3H),2.54(s,1H),2.18-2.09(m,1H),1. 94–1.88 (m, 1H), 1.73 (d, J = 13.0 Hz, 1H), 1.69–1.60 (m, 4H), 1.52 (tt, J = 14.9, 6.2 Hz, 4H), 1.30 (s, 3H), 1.29–1.22 (m, 8H), 1.09–1.02 (m, 4H), 0.92 (d, J = 6.0 Hz, 3H), 0.87 (d, J = 6.1 Hz, 3H). High-resolution mass spectrometry (ESI) + C 75 H 95 Cl2N 11 O 24 Theoretical value [M+H] + m / z 1604.6001, measured value is m / z 1604.5987.

[0261] Example 34: Preparation of compound NV034

[0262]

[0263] In Example 2, NV001 was replaced with NV033, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV034 (23 mg, yield 70%).

[0264] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.82-8.76(m,1H),8.63(s,1H),7.81(s,1H),7.62(s,1H),7.53-7.49(m,1H),7.47(d,J=8.4H z,1H),7.32(d,J=8.4Hz,1H),7.22(d,J=8.4Hz,1H),7.14-7.10(m,1H),6.86(d,J=8.4Hz,1H),6.79 (d,J=8.5Hz,1H),6.54(s,1H),5.75-5.67(m,2H),5.29-5.22(m,2H),5.17-5.13(m,2H),5.11(s,1H ),4.81(s,1H),4.67(q,J=6.6Hz,1H),4.46(d,J=5.5Hz,1H),4.44-4.41(m,1H),4.14-4.03(m,4H), 3.67(d,J=10.8Hz,1H),3.57-3.52(m,2H),3.45-3.40(m,3H),3.30-3.24(m,2H),3.17(s,1H),3.09 -3.03(m,2H),2.93-2.84(m,2H),2.78-2.68(m,1H),2.57(s,3H),2.16-2.08(m,1H),1.94-1.88(m, 1H), 1.77-1.71 (m, 1H), 1.70-1.60 (m, 4H), 1.58-1.48 (m, 2H), 1.48-1.40 (m, 2H), 1.30 (s, 3H), 1.28-1.24 (m, 8H), 1.05 (dd, J = 8.0, 6.0 Hz, 5H), 0.92 (d, J = 6.1 Hz, 3H), 0.87 (d, J = 6.1 Hz, 3H). High-resolution mass spectrometry (ESI) + C 76 H 97 Cl2N 13 O 24 Theoretical value [M+H] + m / z 1646.6219, measured value is m / z 1646.6227.

[0265] Example 35: Preparation of compound NV035

[0266]

[0267] Weigh commercially available vancomycin hydrochloride (75 mg, 0.05 mmol), ({[4-(4-formylphenyl)phenyl]methyl}amino)methane-2-methylpropyl-2-yl ester (compound 6, 31 mg, 0.10 mmol), and DIPEA (43 μL, 0.25 mmol) into a single-necked reaction flask containing 3 mL of DMF and stir. Transfer the reaction system to 50 °C and react until the reaction no longer changes as monitored by analytical RP-HPLC. Then, return the reaction system to room temperature, and slowly add sodium cyanoborohydride (13 mg, 0.2 mmol) dissolved in 300 μL of methanol to the reaction flask. Add an appropriate amount of TFA to the above reaction system to adjust the pH to about 3-4, and continue the reaction for 2 hours to allow the carbon-nitrogen double bonds of the formed Siefer base to be fully reduced to carbon-nitrogen single bonds. Monitor the reaction changes using analytical RP-HPLC. After the reaction was completed, 30 mL of MTBE was added to the reaction system to precipitate the crude product. The supernatant was discarded by centrifugation. Then, the product was deprotected in DCM with 20% TFA at 0 °C for 30 min. After dissolving in water and acetonitrile, the product was directly separated and purified by preparative RP-HPLC. The collected target compound fraction was lyophilized to obtain a white fluffy solid NV035 (20 mg, yield 24%).

[0268] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.72(s,1H),8.59-8.54(m,1H),7.86(s,1H),7.79-7.72(m,4H),7.61-7.53(m,5H),7.4 8(d,J=8.4Hz,1H),7.34(d,J=8.3Hz,1H),7.21(d,J=8.4Hz,1H),7.17(s,1H),6.79(d,J=8.7H z,1H),6.73(d,J=7.9Hz,1H),6.41(d,J=2.4Hz,1H),6.25(d,J=2.4Hz,1H),5.76(d,J=7.6Hz, 1H),5.63(s,1H),5.36(d,J=7.7Hz,1H),5.32-5.29(m,1H),5.22-5.16(m,2H),5.12(s,1H),4. 93(s,1H),4.69(q,J=6.8Hz,1H),4.48-4.46(m,1H),4.44(d,J=5.6Hz,1H),4.26-4.16(m,2H) ,4.13-4.02(m,4H),3.99-3.93(m,1H),3.69(d,J=10.8Hz,1H),3.62-3.57(m,1H),3.32-3.25( m, 2H), 2.64 (s, 3H), 2.21–2.11 (m, 2H), 2.06 (s, 1H), 1.85 (d, J = 13.2 Hz, 1H), 1.73–1.54 (m, 2H), 1.51 (s, 3H), 1.14 (d, J = 6.3 Hz, 3H), 0.91 (d, J = 6.3 Hz, 3H), 0.87 (d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI) + C 80 H 88 Cl2N 10 O 24 Theoretical value [M+H] + m / z 1643.5423, measured value is m / z 1643.5392.

[0269] Example 36: Preparation of compound NV036

[0270]

[0271] In Example 2, NV001 was replaced with NV035, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV036 (11 mg, yield 65%).

[0272] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.73(s,1H),8.58-8.54(m,1H),8.08-8.03(m,1H),7.87-7.84(m,1H),7.76-7.72(m,2H),7.72-7 .69(m,2H),7.58-7.54(m,4H),7.50-7.47(m,1H),7.41(d,J=8.2Hz,2H),7.34(d,J=8.3Hz,1H),7.21(d ,J=8.4Hz,1H),7.17(s,1H),6.79(d,J=8.4Hz,1H),6.73(d,J=8.4Hz,1H),6.41(d,J=2.3Hz,1H),6.25( d,J=2.3Hz,1H),5.76(d,J=7.7Hz,1H),5.64(s,1H),5.36(d,J=7.6Hz,1H),5.30(d,J=4.1Hz,1H),5.19( d,J=3.7Hz,1H),5.18-5.16(m,1H),5.12(s,1H),4.92(s,1H),4.68(q,J=6.6Hz,1H),4.46(d,J=5.6Hz, 1H),4.45-4.40(m,3H),4.25-4.17(m,2H),4.09-4.01(m,2H),3.99-3.94(m,1H),3.69(d,J=10.7Hz,1H) 3.61–3.57 (m, 1H), 3.32–3.26 (m, 2H), 2.63 (s, 3H), 2.19–2.10 (m, 2H), 1.85 (d, J = 13.1 Hz, 1H), 1.71–1.55 (m, 4H), 1.51 (s, 3H), 1.14 (d, J = 6.3 Hz, 3H), 0.92 (d, J = 6.2 Hz, 3H), 0.87 (d, J = 6.3 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 90 Cl2N 12 O 24 Theoretical value [M+H] + m / z 1685.5641, measured value is m / z 1685.5618.

[0273] Example 37: Preparation of compound NV037

[0274]

[0275] The ({[4-(4-formylphenyl)phenyl]methyl}amino)methane-2-methylpropyl-2-yl ester (compound 6) in Example 35 was replaced with ({2-[4-(4-formylphenyl)phenyl]ethyl}amino)methane-2-methylpropyl-2-yl ester (compound 7). The other required raw materials, reagents and preparation steps were the same as in Example 35, and NV037 (23 mg, yield 28%) was obtained.

[0276] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.58(s,1H),8.45-8.39(m,1H),7.71(s,1H),7.57(d,J=7.8Hz,2H),7.51(d,J=7.8Hz,2H),7.44 -7.39(m,3H),7.34(d,J=8.4Hz,1H),7.23(d,J=7.9Hz,2H),7.19(d,J=8.3Hz,1H),7.08(d,J=8.4Hz,1 H),7.02(s,1H),6.64(d,J=8.5Hz,1H),6.58(d,J=8.7Hz,1H),6.28-6.24(m,1H),6.13-6.08(m,1H),5 .61(s,1H),5.49(s,1H),5.22(d,J=7.6Hz,1H),5.16(s,1H),5.06-5.01(m,2H),4.97(s,1H),4.77(s,1 H),4.54(d,J=6.8Hz,1H),4.34-4.31(m,1H),4.29(d,J=5.8Hz,1H),4.12-4.01(m,2H),3.95-3.84(m, 2H),3.58-3.52(m,2H),3.17-3.11(m,3H),3.04-3.00(m,2H),2.96-2.90(m,2H),2.80-2.72(m,3H),2. 58 (s, 1H), 2.51–2.44 (m, 3H), 1.95–1.89 (m, 15H), 1.70 (d, J = 13.1 Hz, 1H), 1.54–1.47 (m, 3H), 1.39–1.32 (m, 3H), 1.09 (s, 1H), 0.99 (d, J = 6.2 Hz, 3H), 0.77 (d, J = 6.0 Hz, 3H), 0.72 (d, J = 5.9 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 90 Cl2N 10 O 24 Theoretical value [M+H] +m / z 1657.5579, measured value is m / z 1657.5582.

[0277] Example 38: Preparation of compound NV038

[0278]

[0279] In Example 2, NV001 was replaced with NV037, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV038 (12 mg, yield 71%).

[0280] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.73(s,1H),8.58-8.54(m,1H),7.86(d,J=1.9Hz,1H),7.72(d,J=8.0Hz,2H),7.64(d,J=8.1Hz,2H) ,7.57-7.54(m,4H),7.50-7.47(m,1H),7.38(d,J=8.1Hz,2H),7.34(d,J=8.3Hz,1H),7.22(d,J=8.3Hz,1H) ,7.18-7.15(m,1H),6.80(d,J=1.9Hz,1H),6.73(d,J=8.4Hz,1H),6.41(d,J=2.3Hz,1H),6.25(d,J=2.3Hz ,1H),5.76(d,J=7.5Hz,1H),5.64(s,1H),5.37(d,J=7.7Hz,1H),5.31(s,1H),5.21-5.17(m,2H),5.12(s,1 H),4.92(s,1H),4.68(q,J=6.7Hz,1H),4.46(d,J=5.6Hz,1H),4.43(d,J=5.6Hz,1H),4.23-4.18(m,2H),4 .08-4.01(m,2H),4.00-3.93(m,1H),3.69(d,J=10.1Hz,1H),3.62-3.57(m,2H),3.43-3.38(m,2H),3.30-3 2.26 (m, 2H), 2.87–2.82 (m, 2H), 2.63 (s, 3H), 2.19–2.10 (m, 2H), 1.85 (d, J = 13.1 Hz, 1H), 1.71–1.55 (m, 2H), 1.51 (s, 3H), 1.23 (s, 1H), 1.14 (d, J = 6.3 Hz, 3H), 0.92 (d, J = 6.1 Hz, 3H), 0.87 (d, J = 6.2 Hz, 3H). High-resolution mass spectrometry (ESI)+ C 82 H 92 Cl2N 12 O 24 Theoretical value [M+H] + m / z 1699.5797, measured value is m / z 1699.5775.

[0281] Example 39: Preparation of compound NV039

[0282]

[0283] Weigh commercially available vancomycin hydrochloride (75 mg, 0.05 mmol) and ({[4-(4-formylphenyl)phenyl]methyl}amino)methane-2-methylpropyl-2-yl ester (compound 6, 46 mg, 0.15 mmol) into a single-necked reaction flask containing 1.5 mL of water and 1.5 mL of acetonitrile and stir. Slowly add sodium cyanoborohydride (32 mg, 0.5 mmol) dissolved in 300 μL of methanol to the reaction system. Then raise the temperature to 70 °C and react until the reaction stops when the analytical RP-HPLC monitoring shows no further change. Subsequently, a large amount of acetonitrile is added to precipitate a white precipitate, which is centrifuged to obtain a white solid crude product. Repeat the above operation twice, and then deprotect the product in DCM with 20% TFA at 0 °C for 30 min. After adding appropriate amounts of water and acetonitrile, separate and purify the product by preparative RP-HPLC, NV039 (18 mg, yield 22%).

[0284] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.54(s,1H),7.84(s,1H),7.80-7.76(m,2H),7.56(d,J=8.0Hz, 4H),7.49-7.46(m,1H),7.35(d,J=8.4Hz,1H),7.22(s,1H),7.16(s,1H),6.81-6.77 (m,1H),6.73(d,J=8.5Hz,1H),6.41(d,J=2.3Hz,1H),6.26(d,J=2.3Hz,1H),5.77(s ,1H),5.62(s,1H),5.28-5.22(m,3H),5.18(s,1H),5.12(s,1H),4.93(s,1H),4.72- 4.63(m,1H),4.44(d,J=5.8Hz,1H),4.19(s,1H),4.09(s,3H),3.67(d,J=10.8Hz,1 H),3.58-3.50(m,2H),3.31-3.23(m,2H),3.16(s,3H),2.84-2.65(m,2H),2.14(s,1 1.93–1.86 (m, 1H), 1.72 (d, J = 13.1 Hz, 1H), 1.68–1.60 (m, 1H), 1.29 (s, 3H), 1.23 (s, 1H), 1.06 (d, J = 6.3 Hz, 3H), 0.99–0.94 (m, 3H), 0.91 (d, J = 6.0 Hz, 3H). High-resolution mass spectrometry (ESI) + C 80 H 88 Cl2N 10 O 24 Theoretical value [M+H] + m / z 1643.5423, measured value is m / z 1643.5403.

[0285] Example 40: Preparation of compound NV040

[0286]

[0287] In Example 2, NV001 was replaced with NV039, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV040 (9 mg, yield 60%).

[0288] 1H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.76(s,1H),8.56(s,1H),7.85(d,J=1.9Hz,1H),7.83-7.78(m,1H),7.76-7.73(m,2H),7 .67-7.59(m,2H),7.54(s,1H),7.49-7.46(m,1H),7.43-7.40(m,2H),7.35(d,J=8.3Hz,1H),7. 21(d,J=8.3Hz,1H),7.16(s,1H),6.78(d,J=8.4Hz,1H),6.73(d,J=8.5Hz,1H),6.41(d,J=2.3H z,1H),6.26(d,J=2.3Hz,1H),5.78(d,J=7.7Hz,1H),5.63(s,1H),5.28-5.21(m,2H),5.17(s,1 H),5.12(s,1H),4.93(s,1H),4.70-4.64(m,1H),4.47-4.41(m,3H),4.18(s,1H),3.68(d,J=10 .8Hz,1H),3.57-3.52(m,2H),3.44-3.40(m,2H),3.30-3.23(m,2H),3.19-3.16(m,1H),2.83-2 0.68 (m, 3H), 2.13 (s, 1H), 1.94–1.87 (m, 1H), 1.73 (d, J = 13.1 Hz, 1H), 1.66–1.59 (m, 1H), 1.30 (s, 3H), 1.23 (s, 1H), 1.06 (d, J = 6.3 Hz, 3H), 1.00–0.96 (m, 3H), 0.91 (d, J = 6.4 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 90 Cl2N 12 O 24 Theoretical value [M+H] + m / z 1685.5641, measured value is m / z 1685.5621.

[0289] Example 41: Preparation of compound NV041

[0290]

[0291] The ({[4-(4-formylphenyl)phenyl]methyl}amino)methane-2-methylpropyl-2-yl ester (compound 6) in Example 39 was replaced with ({2-[4-(4-formylphenyl)phenyl]ethyl}amino)methane-2-methylpropyl-2-yl ester (compound 7). The other required raw materials, reagents and preparation steps were the same as in Example 39, and NV041 (20 mg, yield 24%) was obtained.

[0292] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.74(s,1H),8.54(s,1H),7.86-7.83(m,1H),7.80-7.73(m,2H),7.69(d,J=7.9Hz,2H),7 .62-7.52(m,3H),7.49-7.46(m,1H),7.38(d,J=8.0Hz,2H),7.35(d,J=8.4Hz,1H),7.21(d,J=8 .4Hz,1H),7.16(s,1H),6.78(d,J=8.5Hz,1H),6.73(d,J=8.4Hz,1H),6.41(d,J=2.3Hz,1H),6. 26(d,J=2.3Hz,1H),5.77(s,1H),5.61(s,1H),5.28-5.21(m,3H),5.18(s,1H),5.12(s,1H),4.9 4(s,1H),4.70-4.65(m,1H),4.47-4.42(m,2H),4.19(s,1H),3.67(d,J=10.6Hz,1H),3.58-3.5 2(m,1H),3.54-3.52(m,1H),3.45-3.42(m,1H),3.30-3.24(m,2H),3.18-3.15(m,2H),3.11-3.0 6 (m, 2H), 2.95–2.88 (m, 2H), 2.14 (s, 1H), 1.93–1.87 (m, 1H), 1.73 (d, J = 13.1 Hz, 1H), 1.63 (s, 1H), 1.29 (s, 3H), 1.06 (d, J = 6.3 Hz, 3H), 0.96 (d, J = 6.0 Hz, 3H), 0.90 (d, J = 6.3 Hz, 3H). High-resolution mass spectrometry (ESI) + C 81 H 90 Cl2N 10 O 24 Theoretical value [M+H] + m / z 1657.5579, actual value is m / z 1657.5557.

[0293] Example 42: Preparation of compound NV042

[0294]

[0295] In Example 2, NV001 was replaced with NV041, and the remaining raw materials, reagents and preparation steps were the same as in Example 2, to obtain NV042 (11 mg, yield 65%).

[0296] 1 H NMR (600MHz, DMSO-d6with 20μL D2O)δ8.76(s,1H),8.56(s,1H),7.84(s,1H),7.80-7.76(m,2H),7.70-7.66(m,3H),7.63-7.58( m,2H),7.54(s,1H),7.52-7.49(m,1H),7.49-7.45(m,1H),7.39(d,J=7.9Hz,2H),7.35(d,J=8.3 Hz,1H),7.21(d,J=8.3Hz,1H),7.16(s,1H),6.78(d,J=8.3Hz,1H),6.73(d,J=8.5Hz,1H),6.41( d,J=2.3Hz,1H),6.26(d,J=2.3Hz,1H),5.78(d,J=7.8Hz,1H),5.63(s,1H),5.29-5.21(m,3H),5. 17(s,1H),5.12(s,1H),4.98-4.87(m,1H),4.70-4.64(m,2H),4.45-4.43(m,2H),4.18(s,1H),3 .68(d,J=10.7Hz,1H),3.57-3.53(m,2H),3.43-3.38(m,3H),3.30-3.24(m,2H),3.17(s,1H),2. 87-2.83 (m, 2H), 2.80-2.68 (m, 3H), 2.12 (s, 1H), 1.95-1.86 (m, 2H), 1.75-1.71 (m, 2H), 1.29 (s, 3H), 1.23 (s, 1H), 1.06 (d, J = 6.4 Hz, 3H), 0.97 (d, J = 6.1 Hz, 3H), 0.91 (d, J = 6.3 Hz, 3H). High-resolution mass spectrometry (ESI) + C 82 H 92 Cl2N 12 O 24 Theoretical value [M+H] + m / z 1699.5797, measured value is m / z 1699.5770.

[0297] Biological testing section

[0298] Biological Test 1: In Vitro Antibacterial Activity Test

[0299] In vitro antibacterial activity tests were performed on vancomycin, teicoplanin, and 42 compounds (NV001–NV042) of this invention. The minimum inhibitory concentration (MIC) of the compounds was determined according to the Clinical Laboratory Standards Institute (CLSI) standards for drug susceptibility testing, the technical requirements of WS / T639-2018 antimicrobial susceptibility testing, and the 32nd edition of the CLSI M-100 antimicrobial susceptibility testing implementation standard.

[0300] The tested Gram-positive bacteria strains were: vancomycin-sensitive Staphylococcus aureus (Newman strain, abbreviated as Newman strain), moderately vancomycin-resistant Staphylococcus aureus (Mu50 strain, abbreviated as Mu50 strain), vancomycin-resistant Enterococcus faecium (VRE, VanA phenotype Efm-HS0649 strain, VanM phenotype Efm-HS08257 strain, and VanB phenotype), and vancomycin-resistant Enterococcus faecalis (VRE, VanB phenotype). This invention also tested the antibacterial activity of the compound against four strains of Gram-negative bacteria: *Escherichia coli* (AB1157), *Acinetobacter baumannii* (clinical isolate, Aba strain), *Klebsiella pneumoniae* (clinical isolate, Kpn strain), and *Pseudomonas aeruginosa* (PAO1, Pae strain). These strains were obtained from Renji Hospital, Shanghai.

[0301] In the experiment, 100 μL of the test drug solution at different concentrations (256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125 mg / L) was added to wells 1 through 12 of a sterile 96-well polystyrene plate. Then, 100 μL of the test bacterial culture (200 μL per well) was added to each well, resulting in a final bacterial concentration of approximately 10⁵ CFU / mL. This resulted in final drug concentrations of 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.0625 mg / L, and stepwise dilutions of up to two times the concentration (diluted to additional 96-well polystyrene plates). Each test was performed in replicates. After sealing, the plates were incubated at 35-37°C for 16-24 hours, and the results were then interpreted. The lowest drug concentration that completely inhibits bacterial growth within a small well was defined as the minimum inhibitory concentration (MIC). The results are shown in Tables 1 and 2.

[0302] Table 1

[0303]

[0304]

[0305] Table 2

[0306]

[0307] The above in vitro antibacterial activity studies show that the vancomycin-amino / guanidinyl derivatives and vancomycin-amino / guanidinyl derivatives represented in the embodiments of the present invention exhibit significantly higher antibacterial activity against vancomycin-resistant Staphylococcus aureus and enterococci than vancomycin and vancomycin, with some preferred compounds showing antibacterial activity more than 64 times that of vancomycin or vancomycin. Regarding Gram-negative bacteria, some preferred compounds show 4-16 times greater activity compared to vancomycin and vancomycin, particularly against Escherichia coli. Antibacterial experiments demonstrate that the structural modification strategy of the novel vancomycin-amino / guanidinyl derivatives and vancomycin-amino / guanidinyl derivatives involved in the present invention can significantly enhance their antibacterial activity and shows potential against the inherent resistance of Gram-negative bacteria.

[0308] Biological Test 2: Cytotoxicity Assay

[0309] To investigate the toxicity of the compounds, cytotoxicity experiments were conducted using vancomycin and compounds NV014, NV016, and NV018 prepared in this invention. Two groups of experiments were set up, using HEK-293T: human embryonic kidney cells. Cell viability was measured using the CCK8 (Cell Counting Kit-8) method.

[0310] Cells in the logarithmic growth phase were seeded at an appropriate density (approximately 6000 cells) into 96-well plates, 100 μL per well. After overnight culture, two experimental groups were set up with different concentrations of NV014, NV016, NV018, and vancomycin (25 mg / L, 50 mg / L, and 100 mg / L), 10 μL per well, for 24 h. Each concentration was used in triplicate, with corresponding concentrations of physiological saline as a control and cell-free wells for zeroing. After incubation, 10 μL of CCK8 assay solution was added to each well, and the cells were incubated at 37°C for approximately 1.5 h. The optical density (OD value) at 450 nm was then measured using a Multiskan FC microplate reader. The OD value was converted to cell viability [conversion formula: cell viability value = OD]. C / OD C＝0 ×100, where C represents the concentration. The results are shown in Figure 1.

[0311] The above experiments demonstrate that the compound prepared in this invention has no cytotoxicity to the HEK-293T cell line at the measured concentrations, indicating high safety.

[0312] Biological Test 3: Hemolytic Toxicity Test

[0313] To further test the safety of the compounds of this invention, fresh rat blood was extracted to test the hemolytic effect of the compounds. First, the fresh blood was diluted 4-fold with PBS buffer (Solarbio, China), centrifuged at 3000 rpm for 10 minutes, and repeated 5-6 times until the supernatant was colorless and transparent to collect red blood cells. The washed red blood cells were prepared into an 8% red blood cell suspension with PBS buffer. Compound NV014 was diluted to different concentration gradients (100 mg / L, 50 mg / L, 25 mg / L) and added to the prepared red blood cell suspension, incubated at 37°C for 1 hour. A blank control (red blood cells only) and a positive control (red blood cells containing 0.1% Triton X-100) were set up as references. After 1 hour, the mixture was centrifuged at 3000 rpm for 3 minutes, and the hemolytic effect was directly observed by photographing. The supernatant was then carefully transferred to a 96-well plate to obtain the absorbance at 540 nm. Abt, Abp, and Abn were defined as the absorbance values ​​for the test, positive control, and negative control, respectively. The hemolysis rate is calculated as follows: Hemolysis rate = [(Abt - Abn) / (Abp - Abn)] × 100. The results are shown in Figure 2.

[0314] The above experiments demonstrate that the compound NV014 prepared in this invention has no hemolytic toxicity at a drug concentration of 100 mg / L and has good safety.

[0315] Biological Test 4: Time-to-Kill Curve Experiment

[0316] Compound NV014 was prepared into final concentrations of 1×MIC, 4×MIC, and 8×MIC using appropriate culture media. Staphylococcus aureus MRSA252 or Escherichia coli AB1157 bacterial suspensions were added to each tube to ensure a final bacterial concentration of approximately 10 MIC in each tube. 6 CFU / mL. A bacterial growth control was also included. The bacterial growth control tubes contained only bacterial culture, without any added drug solution. After mixing the liquids in each tube, they were incubated at (35±2)℃ for 24 hours. At 0, 1, 2, 4, 8, and 24 hours of incubation, 100 μL aliquots were taken from each tube for viable cell counting. Viable cell counting method: The 100 μL aliquots were diluted tenfold with sterile physiological saline (10... -1 -10 -7 10 μL of each dilution gradient solution was inoculated onto plates and incubated at (35±2)℃ for 24 hours. The results were then observed. Colony counts were recorded at each inoculation site, with sites ranging from 5 to 50 CFU selected for statistical analysis. The bacterial concentration in the stock solution was calculated using the formula: Stock solution bacterial concentration (CFU / mL) = Plate colony count × Dilution factor × 100. Data were statistically analyzed using log10 CFU / mL. The results were recorded, and a time-sterilization curve was plotted with time on the x-axis and bacterial concentration on the y-axis. The specific results are shown in Figure 3 below.

[0317] As shown in Figure 3(a), for Staphylococcus aureus MRSA252 strain, the compound NV014 prepared in this invention reduced the bacterial count by approximately 1 log10 CFU / mL after treatment with 4×MIC (0.5 μg / mL) and 8×MIC (1 μg / mL) for 24 h, while the 1×MIC treatment showed antibacterial activity. Treatment with 1 μg / mL vancomycin, which is equivalent to the 8×MIC concentration of NV014, showed general antibacterial activity. As shown in Figure 3(b), for Escherichia coli AB1157, the compound NV014 prepared in this invention reduced the bacterial count by approximately 3 log10 CFU / mL after 8 h at 4×MIC (32 μg / mL) and 8×MIC (64 μg / mL), showing a concentration-dependent rapid bactericidal effect, while the 1×MIC treatment showed antibacterial activity. Treatment with 128 μg / mL vancomycin, which is equivalent to 16 × MIC of NV014, showed general antibacterial activity; the above results indicate that the bactericidal performance of the compound NV014 prepared in this invention is superior to that of the negative control vancomycin.

[0318] Biological Test 5: Pharmacokinetics in Mice

[0319] The compound NV014 of this invention and the positive compound vancomycin were selected for in vivo pharmacokinetic experiments in mice. All male mice (CD-1 mice) used were obtained from the Shanghai Laboratory Animal Center of the Chinese Academy of Sciences and were grown to 20-22g under conditions of 18-29℃ and 30-70% humidity.

[0320] Solutions of the aforementioned compounds were injected via the tail vein, and blood samples (from the femoral vein) were collected at seven time points: 0.05 h, 0.25 h, 0.75 h, 2 h, 4 h, 8 h, and 24 h. The corresponding plasma drug concentrations (ng / mL) in the three groups of mice at different time points were detected using LC-MS / MS, and the corresponding half-life (T0) was calculated using the pharmacokinetic software WinNonlin 6.4. 1 / 2 The area under the drug concentration-time curve (AUC), plasma clearance (CL), mean residence time (MRT), and volume of distribution (Vss) calculated when the drug reaches steady state in vivo are shown in Table 3 and Figure 4.

[0321] Table 3. Experimental results of the pharmacokinetic analysis of compound NV014 of the present invention in mice.

[0322]

[0323] Note: AUC last Area under the curve (AUC) of the drug-time curve from the start of dosing to the last point. INF_obs : The area under the pharmacokinetic curve from the start of drug administration to the theoretical extrapolation infinity; CL _obs MRT (Mean Transmission Rate) is the apparent volume of distribution of a drug removed from the body per unit time, i.e., how much drug in a plasma is completely removed per unit time (L / h); INF_obs V: The average residence time from the start of drug administration to the theoretical extrapolation to infinity; SS_obs : The steady-state distribution volume of a drug in the human body (the higher the plasma protein binding, the smaller Vss); obs stands for observed, which is the measured value; INF stands for infinity.

[0324] As shown in Table 3 and Figure 4, the compound NV014 of the present invention has a half-life T 1 / 2 In terms of duration of action, the drug concentration-time curve (AUC) of NV014 is longer than that of vancomycin, and the AUC is 4-5 times larger than that of vancomycin. Regarding plasma clearance (CL), the NV014 compound of this invention exhibits slower clearance than vancomycin, demonstrating superior drug-like properties compared to the positive control compound vancomycin.

[0325] Biological Test 6: In vivo drug efficacy experiment in mice

[0326] To determine the in vivo efficacy of the compounds of this invention, an in vivo protective experiment was conducted using a systemic infection model of drug-resistant MRSA strains to evaluate the in vivo efficacy of this class of glycopeptide antibiotic derivatives.

[0327] In the in vivo efficacy study of MRSA strain (MRSA252) infection, a single-dose administration method was used. Forty 8-week-old female BALB / c mice were randomly divided into 5 groups (NV014-20 mg / kg group, NV014-10 mg / kg group, blank group, control vancomycin-20 mg / kg group, and control vancomycin-10 mg / kg group), with 8 mice in each group. Mice were challenged via tail vein with 1 x 10- MRSA strain infection. 8 CFU containing MRSA252 strain was administered 1 hour later, and the mortality of mice was observed for 7 days. The survival rate was calculated, and it was found that the mice did not die. The results are shown in Figure 5.

[0328] The above experimental results show that the survival rate of mice in the NV014 (10 / 20 mg / kg) group reached 100%, while the survival rate of mice in the vancomycin hydrochloride (20 / 10 mg / kg) group was 75 / 62.5%.

[0329] The above-mentioned in vivo pharmacodynamic studies show that the compound NV014 of the present invention has significant in vivo protective benefits against a systemic infection model of drug-resistant bacteria MRSA252, and its in vivo efficacy is far superior to vancomycin.

[0330] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. A class of glycopeptide antibiotics—amino / guanidine derivatives, characterized in that, It has the structure shown in equation (Ⅰ): Wherein: R0 is selected from -H, -CH3; R1 is selected from -OH, -NH-L-NH2, -NH-L-guanidinyl; R2 is selected from -H, -L-NH2, -L-guanidinyl; R3 is selected from -H, -CH2-NH-L-NH2, -CH2-NH-L-guanidinyl; R4 is selected from -L-NH2, -L-guanidinyl; and at least one of R1, R2, R3, and R4 contains a -NH2 or -guanidinyl structural fragment; L, as a linker, is independently selected from the following groups: substituted or unsubstituted C4-C 20 Straight-chain or branched alkylene, substituted or unsubstituted C4-C 20 Straight-chain or branched alkenyl groups, substituted or unsubstituted C4-C 20 Straight-chain or branched ynylene groups, C4-C 20 Straight-chain or branched imide groups, substituted or unsubstituted C3-C 10 Cycloalkylene, substituted or unsubstituted C6-C 20 A 3-10 membered non-aromatic heterocyclic group containing one or more heteroatoms selected from N, O, and S on a substituted or unsubstituted ring; or a 5-10 membered heterocyclic group containing one or more heteroatoms selected from N, O, and S on a substituted or unsubstituted ring; wherein the substitution refers to being substituted by one or more substituents selected from: halogen, -O-, -OH, -NH2, carbonyl, cyano, C1-C 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Alkylamino, C1-C 10 Alkyl group, C3-C 10 cycloalkyl, halogenated C1-C 10 Alkyl, C2-C6 straight-chain or branched alkynyl, phenylethynyl, trimethylsilylethynyl, pyridyl, phenyl, cyanophenyl, C1-C6 alkylphenyl, trifluoromethylphenyl, chlorophenyl, biphenyl, methyl biphenyl, trifluoromethyl biphenyl, halogen-substituted biphenyl.

2. The glycopeptide antibiotic-amino / guanidine derivative according to claim 1, characterized in that, L, as a linker, is independently selected from the following groups: substituted or unsubstituted C4-C 10 Straight-chain or branched alkylene, substituted or unsubstituted C4-C 10 Straight-chain or branched alkenyl groups, substituted or unsubstituted C4-C 10 Straight-chain or branched ynylene groups, C4-C 10 Straight-chain or branched imide groups, substituted or unsubstituted C3-C 10 Cycloalkylene, substituted or unsubstituted C6-C 14 The arylene group, a 3-10 membered non-aromatic heterocyclic group with one or more heteroatoms selected from N, O, and S on a substituted or unsubstituted ring, or a 5-10 membered heteroarylene group with one or more heteroatoms selected from N, O, and S on a substituted or unsubstituted ring; the substitution refers to being substituted by one or more substituents selected from: halogen, -O-, -OH, -NH2, carbonyl, cyano, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkanoyl, C3-C 10 Cycloalkyl, halogenated C1-C6 alkyl, C2-C6 straight-chain or branched alkynyl, phenylethynyl, trimethylsilylethynyl, pyridyl, phenyl, cyanophenyl, C1-C6 alkylphenyl, trifluoromethylphenyl, chlorophenyl, biphenyl, methyl biphenyl, trifluoromethyl biphenyl, halogen-substituted biphenyl.

3. The glycopeptide antibiotic-amino / guanidine derivative according to claim 2, characterized in that, L, as a linker, is independently selected from the following groups: -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -(CH2)7-, -(CH2)8-, -CO(CH2)2-, -CO(CH2)3-, -(CH2)2-O-(CH2)2-, -(CH2)2-O-(CH2)2-O-(CH2)2-, -CH2-benzene-CH2-, -CH2-biphenyl-CH2-.

4. A pharmaceutically acceptable salt of an amino / guanidine derivative of a glycopeptide antibiotic as described in any one of claims 1 to 3.

5. A method for preparing a glycopeptide antibiotic-amino / guanidine derivative as described in any one of claims 1 to 3, characterized in that, The process includes the following steps: vancomycin hydrochloride or vancomycin hydrochloride undergoes site-selective amide condensation, Mannich reaction or reductive amination reaction, and deprotection reaction to obtain vancomycin or vancomycin derivatives with a terminal primary amino group; guanidinolation reaction is then carried out, and vancomycin derivatives or vancomycin derivatives with a terminal guanidino group are obtained through selective substitution reaction with a guanidinolation reagent.

6. The glycopeptide antibiotic-amino / guanidine derivative according to claim 5, characterized in that, The amide condensation reaction, Mannich reaction, or reductive amination reaction are carried out at -20°C to 70°C.

7. The glycopeptide antibiotic-amino / guanidine derivative according to claim 5, characterized in that, The deprotection reaction is carried out at -10℃ to 10℃; the solvent for the deprotection reaction is a trifluoroacetic acid-water mixture or a trifluoroacetic acid-dichloromethane mixture, wherein the volume content of trifluoroacetic acid in the solvent is 10% to 30%; and the deprotection reaction time is 10 min to 2 h.

8. The glycopeptide antibiotic-amino / guanidine derivative according to claim 5, characterized in that, The guanidinizing agent is selected from 1H-pyrazole-1-formamidinium hydrochloride, N-BOC-1H-pyrazole-1-formamidinium, N,N'-di-BOC-1H-1-guanidinylpyrazole, and N,N'-di-BOC-S-methylisothiourea. The guanidinization reaction is carried out at 20°C to 70°C.

9. A pharmaceutical composition, characterized in that, It comprises a pharmaceutically acceptable salt of a glycopeptide antibiotic-amino / guanidine derivative as described in any one of claims 1 to 3 or a glycopeptide antibiotic-amino / guanidine derivative as described in claim 4.

10. Use of a glycopeptide antibiotic-amino / guanidinium derivative as described in any one of claims 1 to 3, a pharmaceutically acceptable salt of a glycopeptide antibiotic-amino / guanidinium derivative as described in claim 4, or a pharmaceutical composition as described in claim 9 in the preparation of a medicament for treating and / or preventing diseases or conditions associated with Gram-positive and / or Gram-negative bacterial infections.

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