Amanitin derivatives of cyclic peptide toxins, their preparation methods and applications

Abstract

This invention belongs to the field of pharmaceutical preparation technology, specifically relating to a cyclic peptide toxin Amanitin derivative, its preparation method, and its application. Based on the natural Amanitin compound, glycine was replaced with alanine; simultaneously, the eighth amino acid was also replaced. The synthesized cyclic heptapeptide was sequentially oxidized, and proline, phenylalanine, dibutylisobutyric acid, and dihydroxyisoleucine were linked to design and synthesize a series of Amanitin derivatives, namely, products 1 to compounds 12 of this invention, which are 12 cyclic peptide toxin derivatives, and they exhibit good cellular activity.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparation technology, specifically relating to a cyclic peptide toxin Amanitin derivative, its preparation method, and its application. Background Technology

[0002] Antibody-drug conjugates (ADCs) are one of the fastest-growing technologies in cancer treatment, combining the precision of monoclonal antibody (mAbs) with the potency of cytotoxic drugs. This work has made significant contributions to the field of bioconjugation. Amanitin, a bicyclic peptide composed of eight amino acids, is a eukaryotic RNA polymerase II inhibitor that can inhibit cellular transcription and protein synthesis even at extremely low concentrations. When used as a loading medium for ADCs, it has shown promising efficacy in preclinical trials. Currently, there are relatively few derivatives targeting amanitin, and its high hydrophilicity results in poor cell penetration, thus affecting the efficacy of subsequent drugs. Summary of the Invention

[0003] To address the aforementioned problems, this invention provides a cyclic peptide toxin amanitin derivative, its preparation method, and its applications. Based on the structure and function of ADCs, this invention designs and synthesizes a series of amanitin derivatives. In the natural amanitin compound, glycine is replaced with alanine; simultaneously, the eighth amino acid is also replaced. The synthesized cyclic heptapeptide is sequentially oxidized, and proline, phenylalanine, dibutylisobutyric acid, and dihydroxyisoleucine are linked to obtain a series of cyclic peptide toxin derivatives.

[0004] To achieve the above objectives, the technical solution of the present invention is as follows.

[0005] Amanitin derivatives of cyclic peptide toxins having any one of the following structural formulas: .

[0006] A second aspect of this invention provides a method for preparing the aforementioned cyclic peptide toxin Amanitin derivative, comprising the following steps: Compound 8 was subjected to carbon-sulfur cyclization in an iodine-mediated manner to obtain compound 9; Compound 9 was subjected to a deprotection reaction to obtain compound 10; Compound 10 undergoes a deprotection reaction and a condensation reaction to obtain compound 11, wherein compound 11 is any one of product 1 to product 6; Compound 11 was subjected to a deprotection reaction to obtain compound 12, wherein compound 12 is any one of product 7 to product 12; Products 1 to 12 are Amanitin derivatives of the cyclic peptide toxin compound.

[0007] Specifically, the preparation method of the cyclic peptide toxin Amanitin derivative is as follows: Step 1: Fmoc-Hyp, pyridine, and acetyl chloride were mixed with an organic solvent in a molar ratio of 1:1.1:1.1 to carry out an acetylation reaction. The resulting acetylated product, N,N-dimethylpyridine, and ditert-butyl dicarbonate were mixed in a molar ratio of 1:1.1:1.1 to carry out an esterification reaction to obtain compound 1.

[0008] Step 2: Compound 1, N-Fmoc-asparagine, 1-hydroxybenzotriazole, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride were mixed with an organic solvent in a molar ratio of 1:1.1:1.1 to carry out a condensation reaction and a deprotection reaction to obtain compound 2.

[0009] Step 3: Compound 2, N-Fmoc-cysteine, 1-hydroxybenzotriazole, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride are mixed with an organic solvent in a molar ratio of 1:1.1:1.1 to carry out a condensation reaction and a deprotection reaction to obtain compound 3.

[0010] Step 4: Compound 3, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and organic solvent are mixed to carry out condensation and deprotection reactions to obtain compound 4.

[0011] Step 5: Compound 4, N-Fmoc-alanine, 1-hydroxybenzotriazole, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride are mixed with an organic solvent in a molar ratio of 1:1.1:1.1 to carry out a condensation reaction and a deprotection reaction to obtain compound 5.

[0012] Step 6: Compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride are mixed with an organic solvent in a molar ratio of 1:1.1:1.1 to carry out a condensation reaction and a deprotection reaction to obtain compound 6.

[0013] Step 7: The compound 6, N-Fmoc-tryptophan, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent are mixed to carry out a condensation reaction and a deprotection reaction to obtain the compound 7.

[0014] Step 8: Reaction of compound 7 with N-Fmoc-L-isoleucine and removal of the Fmoc protecting group yields white solid compound 8a; reaction of compound 7 with N-Fmoc-proline and removal of the Fmoc protecting group yields white solid compound 8c; reaction of compound 7 with N-Fmoc-phenylalanine and removal of the Fmoc protecting group yields white solid compound 8d; reaction of compound 7 with diaminoisobutyric acid and removal of the Fmoc protecting group yields white solid compound 8e; reaction of compound 7 with dihydroxyisoleucine and removal of the Fmoc protecting group yields white solid compound 8f.

[0015] Step 9: The compound 7 is mixed with m-chloroperoxybenzoic acid and an organic solvent to carry out an oxidation reaction to obtain the compound 8b.

[0016] Step 10: Under a protective atmosphere, the compounds 8a, 8b, 8c, 8d, 8e, 8f and an organic solution of iodine are mixed to perform carbon-sulfur bond cyclization to obtain the compounds 9a, 9b, 9c, 9d, 9e, 9f.

[0017] Step 11: Mix the compounds 9a, 9b, 9c, 9d, 9e, and 9f with an organic solvent to carry out a deprotection reaction to obtain the compounds 10a, 10b, 10c, 10d, 10e, and 10f.

[0018] Step 12: Compounds 10a, 10b, 10c, 10d, 10e, 10f, diisopropylethylamine, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate and an organic solvent are mixed and subjected to deprotection and condensation reactions to obtain compounds 11a, 11b, 11c, 11d, 11e, and 11f.

[0019] Step 13: Deprotection reaction of compounds 11a, 11b, 11c, 11d, 11e, and 11f to obtain the cyclic peptide toxin compound Amanitin derivative.

[0020] Compound 8 is any one of 8a, 8b, 8c, 8d, 8e, and 8f, and its structural formula is shown below: , , , , , .

[0021] Compound 9 is any one of 9a, 9b, 9c, 9d, 9e, and 9f, and its structural formula is shown below: , , , , , .

[0022] Compound 10 is any one of 10a, 10b, 10c, 10d, 10e, and 10f, and its structural formula is shown below: , , , , , .

[0023] In another preferred embodiment, the preparation of compound 9 further includes the introduction of nitrogen gas as a protective atmosphere.

[0024] In another preferred embodiment, the preparation of compounds 9, 10 and 11 further includes purification by silica gel column chromatography after the reaction is completed.

[0025] In another preferred embodiment, the temperature of the deprotection reaction and the condensation reaction is 20°C to 23°C.

[0026] A third aspect of the present invention provides the use of the aforementioned cyclic peptide toxin Amanitin derivative in the preparation of toxin-loaded materials for use as antibody-drug conjugates.

[0027] Compared with the prior art, the present invention has the following beneficial effects.

[0028] This invention, based on the natural amanitin compound, replaces glycine with alanine and also substitutes the eighth amino acid. The synthesized cyclic heptapeptide is sequentially oxidized, linked with proline, phenylalanine, dibutylisobutyric acid, and dihydroxyisoleucine, resulting in the design and synthesis of a series of amanitin derivatives, namely products 1 to 12 of this invention—12 cyclic peptide toxin derivatives. These were used as toxin loading materials for ADCs, and their cellular activity was evaluated. It was found that oxidation and the introduction of derivatives such as dihydroxyisoleucine and phenylalanine improved cellular activity because they improved the lipophilicity of the original compound, giving it better cell penetration and thus enhancing its efficacy. The activity was detected using SKOV3 and HeLa cell lines, employing the CTG activity assay method, aiming to provide a foundation for the design and synthesis of such compounds. Attached Figure Description

[0029] Figure 1The diagrams show the effects of 12 compounds in the HeLa cell line. a represents the effect of product 1, b represents product 7, c represents product 2, d represents product 8, e represents product 3, f represents product 9, g represents product 4, h represents product 10, i represents product 5, j represents product 11, k represents product 6, and l represents product 12.

[0030] Figure 2 The following are effect diagrams of 12 compounds in the SKOV3 cell line: a represents the effect of product 1, b represents the effect of product 7, c represents the effect of product 2, d represents the effect of product 8, e represents the effect of product 3, f represents the effect of product 9, g represents the effect of product 4, h represents the effect of product 10, i represents the effect of product 5, j represents the effect of product 11, k represents the effect of product 6, and l represents the effect of product 12.

[0031] Figure 3 This is a schematic diagram of the structure of the natural amanitin compound.

[0032] Figure 4 This is the hydrogen spectrum of compound 1.

[0033] Figure 5 This is the carbon spectrum of compound 1.

[0034] Figure 6 This is the mass spectrum of compound 1.

[0035] Figure 7 This is the hydrogen spectrum of compound 2.

[0036] Figure 8 This is the carbon spectrum of compound 2.

[0037] Figure 9 This is the mass spectrum of compound 2.

[0038] Figure 10 This is the hydrogen spectrum of compound 3.

[0039] Figure 11 This is the carbon spectrum of compound 3.

[0040] Figure 12 This is the mass spectrum of compound 3.

[0041] Figure 13 This is the hydrogen spectrum of compound 4.

[0042] Figure 14 This is the carbon spectrum of compound 4.

[0043] Figure 15 This is the mass spectrum of compound 4.

[0044] Figure 16 This is the hydrogen spectrum of compound 5.

[0045] Figure 17 This is the carbon spectrum of compound 5.

[0046] Figure 18 This is the mass spectrum of compound 5.

[0047] Figure 19 This is the hydrogen spectrum of compound 6.

[0048] Figure 20 This is the carbon spectrum of compound 6.

[0049] Figure 21 This is the mass spectrum of compound 6.

[0050] Figure 22 This is the hydrogen spectrum of compound 7.

[0051] Figure 23 This is the carbon spectrum of compound 7.

[0052] Figure 24 This is the mass spectrum of compound 7.

[0053] Figure 25 This is the hydrogen spectrum of compound 8a.

[0054] Figure 26 This is the carbon spectrum of compound 8a.

[0055] Figure 27 This is the mass spectrum of compound 8a.

[0056] Figure 28 This is the proton NMR spectrum of compound 8b.

[0057] Figure 29 This is the carbon spectrum of compound 8b.

[0058] Figure 30 This is the mass spectrum of compound 8b.

[0059] Figure 31 This is the proton NMR spectrum of compound 8c.

[0060] Figure 32 This is the carbon spectrum of compound 8c.

[0061] Figure 33 This is the mass spectrum of compound 8c.

[0062] Figure 34 This is the proton NMR spectrum of compound 8d.

[0063] Figure 35 This is the carbon spectrum of compound 8d.

[0064] Figure 36 This is the mass spectrum of compound 8d.

[0065] Figure 37 This is the hydrogen spectrum of compound 8e.

[0066] Figure 38 This is the carbon spectrum of compound 8e.

[0067] Figure 39 This is the mass spectrum of compound 8e.

[0068] Figure 40 This is the hydrogen spectrum of compound 8e.

[0069] Figure 41 This is the carbon spectrum of compound 8e.

[0070] Figure 42 This is the mass spectrum of compound 8e.

[0071] Figure 43 This is the proton NMR spectrum of compound 8f.

[0072] Figure 44 This is the carbon spectrum of compound 8f.

[0073] Figure 45 This is the mass spectrum of compound 8f.

[0074] Figure 46 This is the proton NMR spectrum of compound 9a.

[0075] Figure 47 This is the carbon spectrum of compound 9a.

[0076] Figure 48 This is the mass spectrum of compound 9a.

[0077] Figure 49 This is the proton NMR spectrum of compound 9b.

[0078] Figure 50 This is the carbon spectrum of compound 9b.

[0079] Figure 51 This is the mass spectrum of compound 9b.

[0080] Figure 52 This is the proton NMR spectrum of compound 9c.

[0081] Figure 53 This is the carbon spectrum of compound 9c.

[0082] Figure 54 This is the proton NMR spectrum of compound 9d.

[0083] Figure 55 This is the carbon spectrum of compound 9d.

[0084] Figure 56 This is the mass spectrum of compound 9d.

[0085] Figure 57 This is the proton NMR spectrum of compound 9e.

[0086] Figure 58 This is the carbon spectrum of compound 9e.

[0087] Figure 59 This is the mass spectrum of compound 9e.

[0088] Figure 60 This is the proton NMR spectrum of compound 9f.

[0089] Figure 61 This is the carbon spectrum of compound 9f.

[0090] Figure 62 This is the mass spectrum of compound 9f.

[0091] Figure 63 This is the proton NMR spectrum of compound 10a.

[0092] Figure 64 This is the carbon spectrum of compound 10a.

[0093] Figure 65 This is the mass spectrum of compound 10a.

[0094] Figure 66 This is the proton NMR spectrum of compound 10b.

[0095] Figure 67 This is the carbon spectrum of compound 10b.

[0096] Figure 68 This is the mass spectrum of compound 10b.

[0097] Figure 69 This is the proton NMR spectrum of compound 10c.

[0098] Figure 70 This is the carbon spectrum of compound 10c.

[0099] Figure 71 This is the mass spectrum of compound 10c.

[0100] Figure 72 This is the proton NMR spectrum of compound 10d.

[0101] Figure 73 This is the carbon spectrum of compound 10d.

[0102] Figure 74 This is the mass spectrum of compound 10d.

[0103] Figure 75 This is the proton NMR spectrum of compound 10e.

[0104] Figure 76 This is the carbon spectrum of compound 10e.

[0105] Figure 77 This is the mass spectrum of compound 10e.

[0106] Figure 78 This is the proton NMR spectrum of compound 10f.

[0107] Figure 79 This is the carbon spectrum of compound 10f.

[0108] Figure 80 This is the mass spectrum of compound 10f.

[0109] Figure 81 This is the proton NMR spectrum of compound 11a.

[0110] Figure 82 This is the carbon spectrum of compound 11a.

[0111] Figure 83 This is the mass spectrum of compound 11a.

[0112] Figure 84 This is the proton NMR spectrum of compound 11b.

[0113] Figure 85 This is the carbon spectrum of compound 11b.

[0114] Figure 86 This is the mass spectrum of compound 11b.

[0115] Figure 87 This is the proton NMR spectrum of compound 11c.

[0116] Figure 88 This is the carbon spectrum of compound 11c.

[0117] Figure 89 This is the mass spectrum of compound 11c.

[0118] Figure 90 This is the proton NMR spectrum of compound 11d.

[0119] Figure 91 This is the carbon spectrum of compound 11d.

[0120] Figure 92 This is the mass spectrum of compound 11d.

[0121] Figure 93 This is the proton NMR spectrum of compound 11e.

[0122] Figure 94This is the carbon spectrum of compound 11e.

[0123] Figure 95 This is the mass spectrum of compound 11e.

[0124] Figure 96 This is the proton NMR spectrum of compound 11f.

[0125] Figure 97 This is the carbon spectrum of compound 11f.

[0126] Figure 98 This is the mass spectrum of compound 11f.

[0127] Figure 99 This is the proton NMR spectrum of compound 12a.

[0128] Figure 100 This is the carbon spectrum of compound 12a.

[0129] Figure 101 This is the mass spectrum of compound 12a.

[0130] Figure 102 This is the proton NMR spectrum of compound 12b.

[0131] Figure 103 This is the carbon spectrum of compound 12b.

[0132] Figure 104 This is the mass spectrum of compound 12b.

[0133] Figure 105 This is the proton NMR spectrum of compound 12c.

[0134] Figure 106 This is the carbon spectrum of compound 12c.

[0135] Figure 107 This is the mass spectrum of compound 12c.

[0136] Figure 108 This is the proton NMR spectrum of compound 12d.

[0137] Figure 109 This is the carbon spectrum of compound 12d.

[0138] Figure 110 This is the mass spectrum of compound 12d.

[0139] Figure 111 This is the proton NMR spectrum of compound 12e.

[0140] Figure 112 This is the carbon spectrum of compound 12e.

[0141] Figure 113 This is the mass spectrum of compound 12e.

[0142] Figure 114 This is the proton NMR spectrum of compound 12f.

[0143] Figure 115 This is the carbon spectrum of compound 12f.

[0144] Figure 116 This is the mass spectrum of compound 12f. Detailed Implementation

[0145] The specific embodiments of the present invention are described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention. Unless otherwise specified, the experimental methods described in the embodiments of the present invention are conventional methods.

[0146] Amanitin is a bicyclic peptide composed of eight amino acids, such as... Figure 3 As shown, amanitin is a eukaryotic RNA polymerase II inhibitor that can inhibit cellular transcription and protein synthesis even at extremely low concentrations. When used as a loading medium for ADCs, it has shown promising efficacy in preclinical trials. Currently, there are few derivatives targeting amanitin, and amanitin's high hydrophilicity results in poor cell penetration, thus affecting the efficacy of subsequent drugs.

[0147] This invention, based on the natural amanitin compound, replaces glycine with alanine; simultaneously, it also substitutes the eighth amino acid. The synthesized cyclic heptapeptide is sequentially oxidized, linked with proline, phenylalanine, dibutyl isobutyric acid, and dihydroxyisoleucine, designing and synthesizing a series of amanitin derivatives. This improves the lipophilicity of the original compound, giving it better cell penetration and thus enhancing its efficacy. The principle of the preparation method in this invention is shown in the following formula:

[0148] .

[0149] In the above preparation process, Fmoc is fluorenemethyloxycarbonyl, Ac is acetyl, Trt is triphenylmethyl, and 1~12 represent compounds 1~12 respectively, the same below.

[0150] The following describes a cyclic peptide toxin, Amanitin derivative, its preparation method, and its applications.

[0151] In Example 1, DMAP stands for 4-dimethylaminopyridine.

[0152] The synthesis of compound 1 is based on the following principle: .

[0153] The specific preparation process is as follows: A 2500 mL round-bottom flask was placed with a magnetic stir bar. 35.31 g of 100 mmol of N-Fmoc-4-hydroxyproline was added to the flask, followed by 200 mL of dry dichloromethane. The mixture was stirred and dissolved in an ice-water bath for 5 min. 11.86 g of 150 mmol of pyridine was added using a syringe, and the mixture was stirred for 5 min. 11.70 g of 150 mmol of acetyl chloride was slowly added dropwise using a syringe. The mixture was in an ice-water bath at 0 °C for 30 min, and then reacted at 23 °C for 1 h. After the reaction was complete, the pH was adjusted to 2 with 1 N hydrochloric acid, and the mixture was extracted three times with 500 mL of dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the dichloromethane was removed under reduced pressure to obtain crude product 1.

[0154] Crude product 1 was used directly in the next synthetic reaction without further purification. Crude product 1 was dissolved in 400 mL of tert-butanol, and 1.84 g of 30 mol% DMAP was added. The mixture was stirred at room temperature. A solution containing 43.62 g of (Boc)₂O at 200 mmol was dissolved in 100 mL of tert-butanol and added dropwise to the reaction mixture. The reaction was carried out at room temperature, and TLC was monitored until the starting material was completely consumed. After the reaction was complete, the solvent tert-butanol was removed under reduced pressure to obtain crude product 2. Crude product 2 was dissolved in 150 mL of dichloromethane, and 150 mL of diethylamine was added to remove the Fmoc protecting group. TLC was monitored until the starting material was completely consumed, at which point the reaction was stopped. The mixture of dichloromethane and diethylamine was removed under reduced pressure to obtain crude product 3. Crude product 3 was obtained by column chromatography on a 300-mesh silica gel column, eluting with an ethyl acetate / petroleum ether eluent ratio of 40%–50% to obtain 15.46 g of a yellow oily liquid, which was compound 1, with a yield of 67.5%.

[0155] 1 HNMR: (600MHz, DMSO)δ 5.07 (d, J =2.7Hz, 1H), 3.65(t, J =7.6Hz, 1H), 3.15(dd, J =12.0, 5.Hz, 1H), 2.76(dd, J =12.0,1.5Hz,1H),2.00-1.98(m,5H),1.40(s,9H).

[0156] 13 CNMR(151MHz,DMSO)δ173.1,170.1,80.2,75.0,59.1,52.2,36.4,27.6,20.9.

[0157] HRMS(ESI+)calcd.forC11 H 20 NO4 + (M+H) + :230.1387;found:230.1399. (For example...) Figures 4-6 As shown.

[0158] Example 2: Synthesis of compound 2, the principle of which is as follows: .

[0159] The specific preparation process is as follows: A 2500 mL round-bottom flask was placed with a magnetic stir bar. 65.60 g (0.11 mol) of compound N-Fmoc-asparagine was added to the flask, followed by 500 mL of dry dichloromethane. The mixture was stirred and dissolved, and then in an ice-water bath for 5 min. 12.70 g (0.11 mol) of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 14.90 g (0.11 mol) of 1-hydroxybenzotriazole were added, and the mixture was in an ice-water bath for 30 min. Subsequently, 35.70 g (0.1 mol) of compound 1 was added dropwise, and the mixture was reacted in an ice-water bath for 30 min. The reaction was allowed to proceed overnight at room temperature, and TLC was used to monitor the reaction until the reactants were completely consumed. The organic phase was washed three times with saturated sodium bicarbonate and extracted three times with 1000 mL of dichloromethane. The organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to obtain crude product 1. The unpurified crude product 1 was directly used in the next step of the deprotection reaction. The crude product 1 was dissolved in a 50 wt% diethylamine solution in dichloromethane. The mixture was stirred at 20°C, and the reaction was stopped once the starting material was consumed by TLC. The solvent and diethylamine were removed under reduced pressure, and the mixture was eluted using a 200-300 mesh silica gel column with an ethyl acetate / petroleum ether volume ratio of 50%-60% to give a white solid compound 2, 52.00 g, in 89% yield.

[0160] 1 HNMR(600MHz,DMSO)δ9.27(s,1H),7.31–7.15(m,15H),5.25(s,1H),4.24(t, J =8.1Hz, 1H), 3.79(dd, J =16.9,11.3Hz,2H),3.68(dd, J =11.3,3.8Hz,1H),2.41(dd, J =14.5,10.9Hz,1H),2.34–2.26(m,1H),2.20(d, J =14.7Hz,1H),2.11–2.03(m,1H),2.01(s,3H),1.89(d, J =10.8Hz,2H),1.38(s,9H).

[0161] 13 CNMR(151MHz,DMSO)δ173.1,170.3,169.9,169.6,145.0,128.5,127.4,126.3,80.6,72.7,69.4,58.2,51.5,49.9,40.8,33.9,27.5,20.9.

[0162] HRMS(ESI+)calcd.forC 34 H 40 N3O6 + (M+H) + :586.2912;found:586.2896. (For example...) Figures 7-9 As shown.

[0163] Example 3: Synthesis of compound 3, the principle of which is as follows: .

[0164] Take a 2500 mL round-bottom flask and place a magnetic stir bar inside. Add 64.40 g (0.11 mol) of compound N-Fmoc-cysteine ​​to the flask, add 500 mL of dry dichloromethane, stir to dissolve, and react in an ice-water bath for 5 min. Add 12.70 g (0.11 mol) of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 14.90 g (0.11 mol) of 1-hydroxybenzotriazole, and react in an ice-water bath for 30 min. Then add 70.70 mg (0.11 mol) of compound 2 dropwise, and react in an ice-water bath for 30 min. React overnight at room temperature, monitoring the reaction by TLC until the starting material is consumed. Wash the organic phase with saturated sodium bicarbonate, extract three times with 1000 mL of dichloromethane, combine the organic phases, wash with salt, dry with anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure to obtain the crude product. The unpurified crude product was used directly in the next deprotection reaction. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane, stirred at room temperature, and the reaction was stopped when the starting material was consumed by TLC. The solvent and diethylamine were removed under reduced pressure, and the mixture was eluted using a 300-mesh silica gel column chromatography system with an ethyl acetate / petroleum ether eluent ratio of 75%–80% (v / v), yielding a white solid compound 3, 76.30 g, in 82% yield.

[0165] 1 HNMR(600MHz,DMSO)δ8.73(s,1H),8.25(d, J =7.8Hz,1H),7.34-7.29(m,12H),7.23(dd, J =15.4, 7.2 Hz, 10 Hz), 7.16 (d, J=7.4Hz,8H),5.20(s,1H),4.65(dd, J =13.0,8.0Hz,1H),4.21(t, J =8.1Hz, 1H), 3.74(dt, J =11.6,7.9Hz,2H),3.13(dd, J =7.7, 5.0 Hz, 1H), 2.65 (dd, J =14.8,8.9Hz,1H),2.50-2.48(m,2H),2.25(ddd, J =20.0,11.6,7.1Hz,2H),2.05(ddd, J =13.4,7.8,5.3Hz,1H),1.93(s,3H),1.70(s,2H),1.39(s,9H).

[0166] 13 CNMR(151MHz,DMSO)δ172.8,170.1,169.9,169.6,168.3,144.7,144.5,129.1,128.5,127.9,12 7.4,126.7,126.3,80.7,72.4,69.5,65.9,58.1,53.8,51.7,47.9,38.0,37.1,33.9,27.5,20.8.

[0167] HRMS(ESI+)calcd.forC 34 H 40 N3O6 + (M+H) + :931.4099;found:931.4084. (For example...) Figures 10-12 .

[0168] Example 4: Synthesis of compound 4, the principle of which is as follows: .

[0169] A 2500 mL round-bottom flask was fitted with a magnetic stir bar. 32.70 g (0.11 mol) of compound N-Fmoc-alanine was added to the flask, followed by 500 mL of dry dichloromethane. The mixture was stirred until dissolved and reacted in an ice-water bath for 5 min. Then, 12.70 g (0.11 mol) of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 14.90 g (0.11 mol) of 1-hydroxybenzotriazole were added, and the mixture was reacted in an ice-water bath for 30 min. Subsequently, 0.870 g (0.11 mol) of compound 3 was added dropwise, and the mixture was reacted in an ice-water bath for 30 min. The reaction was allowed to proceed overnight at room temperature, and TLC was used to monitor the reaction until the reactants were completely consumed. The organic phase was washed with saturated sodium bicarbonate, extracted three times with 1000 mL of dichloromethane, and the organic phases were combined. The mixture was washed with salt, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to obtain the crude product. The unpurified crude product was used directly for the next deprotection reaction. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane. The mixture was stirred at room temperature, and the reaction was stopped when the starting material was completely consumed by TLC. The solvent and diethylamine were removed under reduced pressure, and the mixture was eluted by a 200-300 mesh silica gel column with an ethyl acetate / petroleum ether eluent ratio of 0%-0.5% to give a white solid compound 4,68.10 g, with a yield of 69%. 1 HNMR(600MHz,DMSO)δ8.66(s,1H),8.55(d, J =7.7Hz,1H),8.13(s,1H),7.39–7.29(m,12H),7.28–7.21(m,10H),7.20–7.13(m,8H),5.21(s,1H),4.73(d, J =5.6Hz, 1H), 4.53(t, J =5.7Hz, 1H), 4.23(t, J =8.0Hz, 1H), 3.75(dt, J =11.5,8.1Hz,2H),3.13–3.05(m,2H),2.63(dd, J =15.2,8.5Hz,1H),2.56(dd, J =15.1, 4.9 Hz, 1H), 2.40 (d, J =5.9Hz,2H),2.28–2.22(m,1H),2.10–2.03(m,1H),1.93(s,3H),1.86(s,2H),1.41(s,9H). 13CNMR(151MHz,DMSO)δ172.4,170.1,170.0,169.4,169.3,168.2,144.7,144.272,129.1,128.5,128.0,127 .4,126.7,126.3,80.6,72.4,69.5,65.7,58.2,51.7,50.2,47.9,44.5,39.5,37.8,34.5,33.8,27.6,20.7. HRMS(ESI+)calcd.forC 58 H 62 N5O8S + (M+H) + :988.4314;found:988.4307. (For example...) Figures 13-15 .

[0170] Example 5: Synthesis of compound 5, the principle of which is as follows: .

[0171] A 2500 mL round-bottom flask was fitted with a magnetic stir bar. 35.30 g (0.11 mol) of N-Fmoc-isoleucine was added to the flask, followed by 500 mL of dry dichloromethane. The mixture was stirred until dissolved and reacted in an ice-water bath for 5 min. Then, 12.70 g (0.11 mol) of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 14.90 g (0.11 mol) of 1-hydroxybenzotriazole were added, and the mixture was reacted in an ice-water bath for 30 min. Subsequently, 98.70 g (0.10 mol) of compound 4 was added dropwise, and the mixture was reacted in an ice-water bath for 30 min. The reaction was allowed to proceed overnight at room temperature, and TLC was used to monitor the reaction until the reactants were consumed. The organic phase was washed with saturated sodium bicarbonate, extracted three times with 1000 mL of dichloromethane, and the organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to obtain the crude product. The unpurified crude product was used directly for the next deprotection reaction. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane. The mixture was stirred at room temperature, and the reaction was stopped when the starting material was completely consumed by TLC. The solvent and diethylamine were removed under reduced pressure, and the mixture was then subjected to silica gel column chromatography with a gradient elution of 0%–1% methanol / dichloromethane (v / v) to give a white solid compound 5, 80.30 g, in 73% yield.

[0172] 1 HNMR(600MHz,DMSO)δ8.67(s,1H),8.45(d, J =7.1Hz, 1H), 8.16(s, 1H), 8.10(d, J=7.9Hz,1H),7.33–7.30(m,12H),7.25–7.23(m,10H),7.18(d, J =6.7Hz, 8H), 5.19(s, 1H), 4.72(d, J =6.4Hz, 1H), 4.48(d, J =6.4Hz, 1H), 4.21(t, J =7.6Hz,1H),3.76–3.73(m,4H),3.36(s,1H),3.06(d, J =4.2Hz,1H),2.68–2.54(m,2H),2.42(d, J =5.6Hz,1H),2.38–2.28(m,1H),2.23‒2.21(m,1H),2.11–2.01(m,1H),1.91(s,3H),1.69(d, J =30.6Hz,2H),1.41(s,9H),1.12–1.02(m,1H),0.85(d, J =6.4Hz,3H),0.79(t, J =7.0Hz, 3H). 13 CNMR(151MHz,DMSO)δ174.9,170.1,169.9,169.3,169.1,168.6,168.3,144.6,144.2,129.0,128.5,127.9,127.4,126.6,126.3,8 0.6,72.2,69.5,65.7,59.3,58.1,51.6,50.9,47.9,41.9,38.1,34.1,33.8,27.6,23.7,20.7,15.8,11.6.HRMS(ESI+)calcd.forC 64 H 73 N6O9S + (M+H) + :1101.5104;found:1101.5122. (For example...) Figures 16-18 .

[0173] Example 6: Synthesis of compound 6, the principle of which is as follows: .

[0174] A 2500 mL round-bottom flask was fitted with a magnetic stir bar. 32.60 g (0.11 mol) of N-Fmoc-glycine was added to the flask, followed by 500 mL of dry dichloromethane. The mixture was stirred until dissolved and reacted in an ice-water bath for 5 min. Then, 12.70 g (0.11 mol) of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 14.90 g (0.11 mol) of 1-hydroxybenzotriazole were added, and the mixture was reacted in an ice-water bath for 30 min. Subsequently, 110.00 g (0.10 mol) of compound 5 was added dropwise, and the mixture was reacted in an ice-water bath for 30 min. The reaction was allowed to proceed overnight at room temperature, and TLC was used to monitor the reaction until the reactants were consumed. The organic phase was washed with saturated sodium bicarbonate, extracted three times with 1000 mL of dichloromethane, and the organic phases were combined. The mixture was washed with salt, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to obtain the crude product. The unpurified crude product was used directly in the next deprotection reaction. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane. The mixture was stirred at room temperature, and the reaction was stopped when the starting material was completely consumed by TLC. The solvent and diethylamine were removed under reduced pressure, and the mixture was then subjected to silica gel column chromatography with a gradient elution of 0%–1.5% (v / v) methanol / dichloromethane to give a white solid compound 6, 70.60 g, in 61% yield.

[0175] 1HNMR(600MHz, DMSO-d6) δ 10.80 (s, 1H), 8.58 (s, 1H), 8.19 (dd, J = 72.3, 7.7 Hz, 2H), 7.84 (d, J = 35.5 Hz, 1H), 7.53 (dd, J = 39.8, 7.8 Hz, 2H), 7.34 (dd, J = 8.4, 3.6 Hz, 2H), 7.32 (d, J = 8.7 Hz, 8H), 7.23 (d, J = 7.5 Hz, 7H), 7.17 (d, J = 8.2 Hz, 7H), 7.14–7.11 (m, 2H), 7.07 (s, 2H), 6.98 (s, 2H), 5.19 (s, 1H), 4.74 (d, J = 7.1 Hz, 1H), 4.34 (d, J = 6.6 Hz, 1H), 4.23 (d, J = 24.2 Hz, 1H), 4.01 (d, J = 7.2 Hz, 1H), 3.80 (s, 1H), 3.73 (s, 2H), 3.65 (s, 2H), 3.56 (s, 2H), 3.51 (dd, J = 8.8, 4.3 Hz, 1H), 3.13 (s, 2H), 3.04 (d, J = 20.0 Hz, 2H), 2.94 (d, J = 7.6 Hz, 2H), 2.75 (d, J = 5.6 Hz, 1H), 2.64 (dd, J = 15.3, 5.9 Hz, 1H), 2.58 (s, 1H), 2.41–2.34 (m, 1H), 2.21 (s, 1H), 2.07 (s, 1H), 1.99 (s, 1H), 1.91 (s, 2H), 1.70 (s, 1H), 1.41 (s, 8H), 1.26 (s, 1H), 1.19 (s, 2H), 1.10 (s, 1H), 1.03 (s, 2H), 0.92 (s, 1H), 0.78 (s, 5H). 13CNMR(151MHz, DMSO) δ 189.5, 176.1, 175.0, 172.1, 170.6, 169.6, 145.2, 144.7, 136.8, 136.6, 129.6, 129.1, 128.4, 127.9, 127.1, 126.8, 124.2, 124.1, 121.3, 118.8, 118.8, 118.7, 111.8, 110.5, 81.1, 73.3, 70.0, 60.3, 58.6, 57.3, 55.8, 55.6, 55.3, 52.1, 51.8, 43.5, 37.4, 31.4, 31.3, 28.1, 27.8, 25.5, 21.2, 18.4, 15.8, 14.9, 14.4, 11.5. HRMS(ESI) m / z calculated for C78H87N9O11S [M + H]+: 1358.6146; found: 1358.5347. As Figures 19-21 .

[0176] Example 7: Synthesis of compounds 7 and 8, the principle of which is as follows: .

[0177] A 2500 mL round-bottom flask was fitted with a magnetic stir bar. 68.90 g (0.10 mol) of N-Fmoc-tryptophan was added to the flask, followed by 500 mL of dry dichloromethane. The mixture was stirred until dissolved and reacted in an ice-water bath for 5 min. Then, 12.70 g (0.11 mol) of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and 14.90 g (0.11 mol) of 1-hydroxybenzotriazole were added, and the mixture was reacted in an ice-water bath for 30 min. Subsequently, 143.80 g (0.11 mol) of compound 6 was added dropwise, and the mixture was reacted in an ice-water bath for 30 min. The reaction was allowed to proceed overnight at room temperature, and TLC was used to monitor the reaction until the reactants were consumed. The organic phase was washed with saturated sodium bicarbonate, extracted three times with 1000 mL of dichloromethane, and the organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to obtain the crude product. The unpurified crude product was used directly in the next deprotection reaction. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane. The mixture was stirred at room temperature, and the reaction was stopped when the starting material was completely consumed by TLC. The solvent and diethylamine were removed under reduced pressure, and the product was purified by silica gel column chromatography (200-300 mesh, methanol / dichloromethane = 0%-2% v / v) to give compound 7, a white solid, 100.00 g, in 69% yield.

[0178] 1HNMR(600MHz,DMSO-d6)δ10.73(s,1H),8.56(s,1H),8.24(d,J=7.9Hz,1H),8.14(s,1H),8.10(d,J=7.4Hz,1H),7.94(d,J=7.9Hz,1H),7.88(d,J=7.6Hz,2H),7.85(d,J=8.3Hz,1H),7.75–7.70(m,2H),7.57(d,J=7.9Hz,1H),7.41(d,J=7.8Hz,2H),7.31(t,J=5.4Hz,12H),7.25–7.20(m,7H),7.17(d,J=3.6Hz,8H),7.12(d,J=7.5Hz,1H),7.03(t,J=7.8Hz,1H),6.95(t,J=7.3Hz,1H),5.19(s,1H),4.74(q,J=7.3Hz,1H),4.61(s,1H),4.38–4.31(m,2H),4.28–4.18(m,4H),3.91–3.85(m,1H),3.77–3.70(m,3H),3.15(dd,J=14.6,4.9Hz,1H),3.00(dd,J=14.8,8.6Hz,1H),2.64(dd,J=15.4,5.9Hz,1H),2.56–2.47(m,6H),2.43–2.33(m,2H),2.21(d,J=17.4Hz,1H),2.10–2.04(m,1H),1.99(s,1H),1.91(d,J=14.6Hz,2H),1.69(s,1H),1.41(s,9H),1.25(d,J=6.8Hz,2H),1.17(t,J=7.1Hz,3H),1.04(dq,J=14.2,7.4Hz,2H),0.84(qd,J=8.9,7.7,4.6Hz,2H),0.79–0.75(m,6H),0.71(d,J=6.9Hz,3H)。13CNMR(151MHz,DMSO)δ172.8,172.3,172.2,172.1,171.1,170.6,170.4,169.8,169.6,169.0,168.9,168.8,156.2,145.2,144. 8,144.4,144.3,143.1,141.2,139.9,137.9,136.6,129.6,129.4,129.1,128.5,128.1,127.9,127.7,127.6,127.1,126.8,125.8 ,124.7,124.1,121.8,121.3,120.5,120.5,118.8,118.7,111.7,110.4,110.0,81.1,72.7,70.0,66.2,60.2,58.7,57.1,55.3,54.0,52.2,51.7,50.5,48.6,48.3,47.2,42.6,38.6,37.4,34.5,34.4,28.1,28.0,27.9,24.8,21.2,18.6,18.5,15.8,14.5,11.5. HRMS(ESI)m / z calculated for C101H112N10O14S[M]+:1716.8080;found:1716.6033. (Example) Figures 22-24 .

[0179] The synthesis principle of compound 8a is as follows: .

[0180] 18.85 g (14.0 mmol) of compound 7, 25 g (15.43 mmol) of N-Fmoc-L-isoleucine, and 2.08 g (15.43 mmol) of 1-hydroxybenzotriazole were added to 80 mL of dry CH2Cl2, and the temperature was maintained at 0 °C. At the same temperature, 2.96 g (15.43 mmol) of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride was gradually added, and the mixture was heated to 23 °C and stirred for 12 h. The reaction was then terminated with 0.5 N, 2 × 80 mL of HCl aqueous solution, and washed with 2 × 80 mL of saturated Na2CO3 aqueous solution. After separation of the organic phase, the mixture was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography using a gradient elution of dichloromethane and methanol (98:2 v / v) to give 20.15 g (11.22 mmol) of white solid compound 8a, in 80% yield, with a melting point of 153–154 °C. 1H NMR (600 MHz, DMSO-d6) δ 10.73 (s, 1H), 8.56 (s, 1H), 8.24 (d, J = 7.9 Hz, 1H), 8.14 (s, 1H), 8.10 (d, J = 7.4 Hz, 1H), 7.94 (d, J= 7.9 Hz, 1H), 7.88 (d, J = 7.6 Hz, 2H), 7.85 (d, J = 8.3 Hz, 1H), 7.75 –7.70 (m, 2H), 7.57 (d, J = 7.9 Hz, 1H), 7.41 (d, J = 7.8 Hz, 2H), 7.31 (t, J= 5.4 Hz, 12H), 7.25 – 7.20 (m, 7H), 7.17 (d, J = 3.6 Hz, 8H), 7.12 (d, J =7.5 Hz, 1H), 7.03 (t, J = 7.8 Hz, 1H), 6.95 (t, J = 7.3 Hz, 1H), 5.19 (s,1H), 4.74 (q, J = 7.3 Hz, 1H), 4.61 (s, 1H), 4.38 – 4.31 (m, 2H), 4.28 – 4.18(m, 4H), 3.91 – 3.85 (m, 1H), 3.77 – 3.70 (m, 3H), 3.15 (dd, J = 14.6, 4.9Hz, 1H), 3.00 (dd, J = 14.8, 8.6 Hz, 1H), 2.64 (dd, J = 15.4, 5.9 Hz, 1H), 2.56 – 2.47 (m, 6H), 2.43 – 2.33 (m, 2H), 2.21 (d, J = 17.4 Hz, 1H), 2.10 – 2.04 (m, 1H), 1.99 (s, 1H), 1.91 (d, J = 14.6 Hz, 2H), 1.69 (s, 1H), 1.41 (s, 9H), 1.25 (d, J = 6.8 Hz, 2H), 1.17 (t, J = 7.1 Hz, 3H), 1.04 (dq, J = 14.2, 7.4 Hz, 2H), 0.84 (qd, J = 8.9, 7.7, 4.6 Hz, 2H), 0.79 – 0.75 (m, 6H), 0.71 (d, J = 6.9 Hz, 3H). 13C NMR (151 MHz, DMSO-d6) δ 172.83, 172.16, 172.12, 170.60, 170.44, 169.85, 169.58, 168.86, 168.78, 145.21, 144.76, 144.27, 143.11, 141.21, 139.94, 137.95, 136.56, 129.60, 129.39, 129.09, 128.99, 128.46, 128.09, 127.92, 127.88, 127.74, 127.55, 127.12, 126.77, 125.76, 124.10, 121.82, 121.27, 120.52, 120.46, 118.83, 118.68, 111.71, 110.39, 110.04, 81.15, 72.71, 70.03, 66.21, 58.65, 57.06, 53.96, 52.17, 51.68, 48.62, 48.28, 47.19, 42.63, 38.61, 37.42, 34.53, 34.35, 28.10, 27.93, 24.80, 21.18, 18.61, 18.50, 15.84, 14.39, 11.47. HRMS (ESI) m / z calculated for C78H87N9O11S [M+H]+: 1358.6146; found: 1358.5347. Such as. Figures 25-27 .

[0181] Following the synthesis method of compound 8a, 143.80 g and 0.11 mol of compound 8a and 35.7 g and 0.10 mol of m-chlorobenzoic acid were reacted to produce a white solid compound 8b, 104.80 g and 78%. 1 HNMR(600MHz,DMSO-d6)δ10.76(d,J=18.3Hz,1H),8.67(d,J=47.2Hz,1H),8.27(s,1H),7.96(s,2H),7.89(d,J=7.1Hz,4H),7.75(d,J=30.6Hz, 3H),7.56(s,1H),7.43–7.12(m,23H),7.10–6.90(m,3H),5.27(d,J=78. 6Hz,1H),4.55(s,3H),4.45–4.30(m,3H),4.23(s,4H),3.78(d,J=106.5 Hz,5H),3.56(s,1H),3.44(dt,J=14.4,7.1Hz,1H),3.09(s,2H),2.99(s ,1H),2.90(s,2H),2.74(s,2H),2.56(s,2H),2.31(s,1H),2.02(d,J=37 .2Hz,3H),1.69(d,J=31.9Hz,3H),1.39(s,9H),1.24(d,J=4.1Hz,6H),1.16–0.98(m,6H),0.80(d,J=7.1Hz,14H),0.65(dd,J=52.0,6.6Hz,3H). 13CNMR(151MHz,DMSO)δ170.5,168.7,145.2,143.1,139.9,137.9,129.4,129.0,127.9,127.8,126.7,121.8,12 0.5,110.2,81.2,70.0,60.5,60.2,58.4,55.4,52.9,52.5,49.6,48.3,46.8,43.7,28.1,21.2,15.9,12.0,11.4. HRMS(ESI)m / zcalculatedforC82H96N10O15S[M+H]+:1493.6777,found:1493.1962. like Figures 28-30 .

[0182] Following the synthetic method of compound 8a, 143.80 g and 0.11 mol of compound 7 and 34.80 g and 0.10 mol of N-Fmoc-proline were reacted, and the Fmoc protecting group was removed to give compound 8c, a white solid, 100.80 g, 77%.

[0183] 1 HNMR(600 MHz, DMSO-d6) δ 10.75 (d, J = 57.0 Hz, 1H), 8.63 (d, J = 7.8 Hz, 1H), 8.34 (s, 1H), 8.23–8.09 (m, 2H), 8.02 (d, J = 30.2 Hz, 1H), 7.91 (d, J = 7.6 Hz, 4H), 7.67 (d, J = 7.8 Hz, 3H), 7.59 (d, J = 34.3 Hz, 2H), 7.43 (t, J = 7.4 Hz, 4H), 7.30 (s, 14H), 7.23 (d, J = 7.1 Hz, 8H), 7.17 (d, J = 9.8 Hz, 7H), 7.07–6.91 (m, 2H), 5.18 (s, 1H), 4.73 (s, 1H), 4.53 (s, 1H), 4.40–4.33 (m, 2H), 4.29 (d, J = 2.6 Hz, 4H), 4.21 (d, J = 7.1 Hz, 3H), 4.00 (s, 1H), 3.72 (s, 3H), 3.17 (s, 1H), 3.04 (s, 1H), 2.62 (s, 1H), 2.37 (s, 2H), 2.29–2.17 (m, 2H), 2.07 (s, 1H), 2.00 (d, J = 13.2 Hz, 1H), 1.89 (dd, J = 19.8, 4.6 Hz, 5H), 1.71 (s, 3H), 1.40 (d, J = 9.5 Hz, 8H), 1.29–1.23 (m, 3H), 1.17 (d, J = 10.3 Hz, 3H), 0.76 (s, 3H). 13CNMR(151 MHz, DMSO) δ 172.1, 170.6, 170.4, 169.8, 169.5, 168.7, 145.2, 144.7, 143.1, 139.9, 137.9, 136.5, 129.6, 129.4, 129.1, 128.5, 127.9, 127.9, 127.8, 127.6, 127.1, 126.8, 125.7, 121.8, 120.5, 118.9, 118.6, 111.7, 110.2, 81.1, 72.7, 70.0, 66.1, 60.2, 58.6, 57.0, 55.4, 52.1, 51.5, 49.1, 48.5, 48.2, 38.5, 37.4, 37.4, 34.5, 34.3, 30.5, 28.1, 27.9, 25.9, 24.7, 21.2, 18.5, 15.8, 14.6, 11.5. HRMS(ESI) m / z calculated for C97H104N10O14S [M+Na]+: 1428.6540; found: 1428.5728. As Figures 31-33 .

[0184] Following the synthetic method of compound 8a, 143.80 g and 0.11 mol of compound 7 and 44.80 g and 0.10 mol of N-Fmoc-phenylalanine were reacted, and the Fmoc protecting group was removed to give a white solid compound 8d, 111.80 g, 79%.

[0185] 11H NMR (600 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.26 (d, J = 128.7 Hz, 3H), 7.98 (d, J = 8.4 Hz, 1H), 7.97–7.82 (m, 4H), 7.71 (d, J = 8.3 Hz, 1H), 7.63 (d, J = 9.0 Hz, 2H), 7.61–7.47 (m, 4H), 7.41 (s, 4H), 7.35–7.08 (m, 21H), 7.01 (d, J = 49.5 Hz, 3H), 4.58 (d, J = 25.5 Hz, 1H), 4.26 (s, 1H), 4.19 (s, 2H), 4.13 (s, 3H), 4.04 (d, J = 7.1 Hz, 2H), 3.75 (d, J = 5.8 Hz, 2H), 3.44 (s, 1H), 3.20 (s, 1H), 3.10 (s, 1H), 2.99 (d, J = 14.1 Hz, 3H), 2.95–2.85 (m, 1H), 2.78–2.74 (m, 1H), 1.99 (s, 3H), 1.90 (s, 1H), 1.41 (s, 1H), 1.27 (d, J = 21.8 Hz, 2H), 1.18 (s, 3H), 1.13 (s, 3H), 1.01 (s, 2H), 0.82 (s, 6H). 13C NMR (151 MHz, DMSO) δ 172.1, 171.8, 170.8, 170.7, 168.6, 156.2, 144.3, 144.2, 141.1, 138.6, 136.5, 129.7, 129.7, 129.6, 129.6, 129.1, 128.6, 128.4, 128.3, 128.1, 127.9, 127.7, 127.5, 126.6, 125.8, 125.7, 124.8, 124.1, 121.4, 121.3, 120.5, 119.5, 118.8, 118.7, 111.7, 110.3, 110.1, 66.1, 60.2, 56.6, 53.9, 52.5, 47.1, 42.4, 42.0, 41.5, 37.9, 37.4, 28.1, 27.9, 24.5, 21.2, 15.7, 15.0, 14.6, 14.5, 13.3, 13.2, 11.4. HRMS (ESI) m / z calculated for C102H106N10O14S [M]+: 1716.7612; found: 1716.6033. As Figures 34-36 .

[0186] Following the synthetic method of compound 8a, 143.80 g and 0.11 mol of compound 7 and 34.80 g and 0.10 mol of diaminoisobutyric acid were reacted, and the Fmoc protecting group was removed to give a white solid compound 8e, 90.80 g, 73%. 1 HNMR(600MHz,DMSO-d6)δ10.80(s,1H),8.26(d,J=128.7Hz,3H),7.98(d,J=8.4Hz,1H),7.97–7.82(m,4H),7.71(d,J=8.3Hz,1H),7.63(d,J =9.0Hz,2H),7.61–7.47(m,4H),7.41(s,4H),7.35–7.08(m,21H),7.01(d,J=49.5Hz,3H),4.58(d,J=25.5Hz,1H),4.26(s,1H),4.19(s,2H) ,4.13(s,3H),4.04(d,J=7.1Hz,2H),3.75(d,J=5.8Hz,2H),3.44(s,1H),3.20(s,1H),3.10(s,1H),2.99(d,J=14.1Hz,3H),2.95–2.85(m,1 H),2.78–2.74(m,1H),1.99(s,3H),1.90(s,1H),1.41(s,1H),1.27(d,J=21.8Hz,2H),1.18(s,3H),1.13(s,3H),1.01(s,2H),0.82(s,6H). 13CNMR(151MHz,DMSO)δ172.1,171.8,170.8,170.7,168.6,156.2,144.3,144.2,141.1,138.6,136.5,12 9.7,129.7,129.6,129.6,129.1,128.6,128.4,128.3,128.1,127.9,127.7,127.5,126.6,125.8,125.7, 124.8,124.1,121.4,121.3,120.5,119.5,118.8,118.7,111.7,110.3,110.1,66.1,60.2,56.6,53.9,52.5,47.1,42.4,42.0,41.5,37.9,37.4,28.1,27.9,24.5,21.2,15.7,15.0,14.6,14.5,13.3,13.2,11.4. HRMS(ESI)m / z calculated for C102H106N10O14S[M]+:1716.7612;found:1716.6033. (The last part is a repetition of the first and can be omitted.) Figures 37-39.

[0187] Following the synthetic method for compound 8a, 143.80 g and 0.11 mol of compound 7 and 34.80 g and 0.10 mol of diaminoisobutyric acid were reacted, and the Fmoc protecting group was removed to give a white solid compound 8e, 84.80 g, 72%. ¹H NMR (600 MHz, DMSO) δ 10.81, 8.64, 8.36, 8.35, 8.26, 8.25, 8.19, 8.18, 7.91, 7.90, 7.88, 7.85, 7.84, 7.82, 7.81, 7.62, 7.58, 7.47, 7.43, 7.42, 7.41, 7.36, 7.35, 7.34, 7.30, 7.24, 7.22, 7.17, 7.16, 7.12, 7.1 0,7.06,7.05,7.04,6.96,6.95,4.73,4.54,4.37,4.36,4.33,4.32,4.24,4.20,4.18,4.17,4.04,4.03,3.81,3.74,3.72,3.64,3.64,3.18,2.62,2.60,2.35,2.21,1.99,1.98,1.90,1.40,1.39,1.31,1.30,1 .29,1.28,1.24,1.19,1.18,1.11,1.04,0.91,0.90,0.84,0.83,0.76,0.76.13CNMR(151MHz,DMSO)δ19 0.5,172.1,171.1,171.0,170.6,169.8,169.3,168.9,167.8,161.8,161.0,145.2,144.7,144.0,141.5 ,129.6,129.1,128.5,128.1,127.9,127.6,127.1,126.8,125.3,124.7,120.6,119.3,81.1,72.7,70. 0,67.8,66.1,64.0,61.0,58.6,57.0,55.4,51.5,47.9,40.6,34.3,30.9,28.1,27.9,21.2,15.8,11.4. HRMS(ESI)m / zcalculatedforC 78 H 89 N 10 O 14 S[M+Na]+:1389.6560;found:1389.5070. (Example) Figures 40-42 .

[0188] Following the synthetic method of compound 8a, 143.80 g and 0.11 mol of compound 7 and 36.60 g and 0.10 mol of dihydroxyisoleucine were reacted, and the Fmoc protecting group was removed to give a white solid compound 8f, 88.80 g, 75%.

[0189] 1 HNMR(600MHz,DMSO-d6)δ11.19(s,1H),8.63(s,1H),8.42(s,1H),8.28(s,1H),8 .10(d,J=34.1Hz,2H),7.92(d,J=35.9Hz,4H),7.74(s,3H),7.44(d,J=30.8Hz,3H ),7.37–7.23(m,9H),7.17(t,J=8.1Hz,11H),6.99(s,1H),5.18(s,1H),4.90(s,1 H),4.71(d,J=42.2Hz,2H),4.46–4.30(m,2H),4.21(d,J=6.1Hz,3H),4.13–4.05( m,1H),3.88(d,J=15.7Hz,2H),3.70(d,J=18.8Hz,4H),3.35–3.33(m,2H),3.17( s,1H),3.05–2.91(m,2H),2.88(d,J=2.5Hz,1H),2.73(s,1H),2.61(d,J=16.8Hz, 2H),2.27(s,1H),2.04(s,1H),2.01–1.84(m,12H),1.53(s,1H),1.37(s,9H),1.2 8(d,J=10.7Hz,1H),1.21–1.08(m,3H),0.89–0.73(m,9H),0.65(d,J=7.1Hz,2H). 13CNMR(151MHz,DMSO)δ175.4,174.5,173.6,172.5,156.8,144.2,144.2,141.2,131.3,130.5,129.5,129.0,128.1,127.9,127.5,125 .7,125.7,120.6,86.1,80.8,66.4,66.3,62.3,60.4,55.4,55.2,53.4,47.1,47.1,35.6,34.6,30.5,28.0,25.5,21.5,21.0,13.5,8.2. HRMS(ESI) m / z calculated for C103H112N10O18S[M]+: 1808.7877; found: 1817.7610. (For example...) Figures 43-45 .

[0190] Example 9: Synthesis of compounds 9a-9f, the principle of which is as follows: .

[0191] A 2500 mL round-bottom flask was fitted with a magnetic stirrer, and the gas was purged three times using the Schlenk technique. Under N2 atmosphere, 57.00 g of 0.03 mol of compound 8a was added to the flask, followed by 500 mL of dry N,N-dimethylformamide, and stirred until dissolved. The mixture was stirred at 20°C. A 2 mg / mL solution of elemental iodine in DMF was prepared, and 4 equiv of iodine solution was slowly added dropwise to the reaction flask using a syringe. The reaction was carried out at 20°C for 2 h. The reaction was then stopped by adding excess sodium thiosulfate solution to quench the reaction. DMF was removed under reduced pressure, and the mixture was extracted three times with 2000 mL of ethyl acetate. The organic phases were combined, washed with salt, dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure to obtain the crude product. The crude product was eluted using a 300-mesh silica gel column with a methanol / dichloromethane eluent of 0%–2% (v / v) to give 30.60 g of a white solid, compound 9a, in 62% yield.

[0192] 1HNMR(600MHz,DMSO-d6)δ11.12(s,1H),8.56(s,2H),8.27–8.23(m,1H),8.04(d,J=7.6Hz,1H),8.00(d,J=8.6Hz,1H),7.86(d,J=5.8Hz,1H),7.69(d,J=7.9Hz,1H),7.25(dt,J=15.3,8.0Hz,9H),7.17(d,J=6.7Hz,8H),7.16–7.08(m,2H),7.00(t,J=7.5Hz,1H),5.85(s,2H),5.55(s,1H),5.19(s,1H),4.69(d,J=7.2Hz,1H),4.31(t,J=6.5Hz,1H),4.22(t,J=8.2Hz,1H),4.17(td,J=9.7,8.4,3.1Hz,1H),4.02(d,J=5.7Hz,5H),3.87(dd,J=15.3,6.0Hz,1H),3.70(d,J=21.4Hz,2H),3.59(dd,J=15.2,4.6Hz,1H),3.38–3.35(m,1H),3.19(d,J=4.4Hz,9H),3.05–2.99(m,1H),2.95(d,J=4.4Hz,1H),2.82–2.77(m,1H),2.57(d,J=6.5Hz,2H),2.26(d,J=9.7Hz,1H),2.06(d,J=8.0Hz,1H),1.98(d,J=12.1Hz,1H),1.93(d,J=23.0Hz,3H),1.39(s,10H),1.33–1.23(m,3H),1.23–1.17(m,4H),1.10(s,1H),0.87(t,J=7.1Hz,6H),0.58(d,J=6.9Hz,2H),0.51(t,J=7.4Hz,2H)。13CNMR(151MHz,DMSO)δ172.9,172.3,170.9,170.5,170.4,170.0,169.7,168.7,162.8,145.2,144.4,143.0,141.0,140 .9,139.9,137.9,137.7,135.9,133.7,130.0,129.4,129.0,129.0,128.2,127.9,127.9,127.8,127.3,126.9,126.7,12 4.4,122.6,122.5,121.8,121.7,121.1,120.5,118.9,110.2,81.2,72.9,70.0,62.3,60.2,58.5,55.4,54.3,53.1,52.3,51.9,49.4,48.3,38.1,37.6,36.3,34.4,31.3,28.0,27.7,24.9,22.9,21.2,18.8,15.8,15.8,15.5,14.5,12.0,11.4. HRMS(ESI)m / zcalculatedforC82H96N10O14S[M]+:1471.6828;found:1471.5237. like. Figures 46-48 .

[0193] Following the synthetic method of compound 9a, compound 9b, a white solid, was obtained in 34.40 g, with a yield of 74%. 1HNMR(600MHz,DMSO-d6)δ10.76(d,J=18.3Hz,1H),8.67(d,J=47.2Hz,1H),8.27(s,1H),7.96(s,2H),7.89(d,J=7.1Hz,4H),7.75(d,J=30.6Hz, 3H),7.56(s,1H),7.43–7.12(m,23H),7.10–6.90(m,3H),5.27(d,J=78. 6Hz,1H),4.55(s,3H),4.45–4.30(m,3H),4.23(s,4H),3.78(d,J=106.5 Hz,5H),3.56(s,1H),3.44(dt,J=14.4,7.1Hz,1H),3.09(s,2H),2.99(s ,1H),2.90(s,2H),2.74(s,2H),2.56(s,2H),2.31(s,1H),2.02(d,J=37 .2Hz,3H),1.69(d,J=31.9Hz,3H),1.39(s,9H),1.24(d,J=4.1Hz,6H),1.16–0.98(m,6H),0.80(d,J=7.1Hz,14H),0.65(dd,J=52.0,6.6Hz,3H). 13CNMR(151MHz,DMSO)δ170.5,168.7,145.2,143.1,139.9,137.9,129.4,129.0,127.9,127.8,126.7,121.8,12 0.5,110.2,81.2,70.0,60.5,60.2,58.4,55.4,52.9,52.5,49.6,48.3,46.8,43.7,28.1,21.2,15.9,12.0,11.4. HRMS(ESI)m / zcalculatedforC82H96N10O15S[M+H]+:1493.6777,found:1493.1962. like Figures 49-51 .

[0194] Following the synthetic method of compound 9a, compound 9c, a white solid, was obtained in 35.80 g, with a yield of 73%. 1HNMR(600MHz,DMSO-d6)δ11.26(d,J=106.4Hz,1H),8.61(s,1H),8.21(s,1H ),8.12(d,J=26.6Hz,2H),7.93(d,J=35.5Hz,2H),7.87(s,2H),7.69(s,2H) ,7.51(s,1H),7.42(s,2H),7.35(s,2H),7.24(s,4H),7.16(d,J=7.9Hz,6H) ,7.00(s,1H),5.18(s,1H),4.75(d,J=105.4Hz,2H),4.23(d,J=52.2Hz,5H) ,4.03(d,J=15.4Hz,2H),3.94(d,J=29.2Hz,2H),3.66(s,2H),3.18(d,J=4. 8Hz,1H),3.00(s,1H),2.90(s,1H),2.74(d,J=2.0Hz,1H),2.59(d,J=40.8H z,2H),2.26(s,2H),2.03(d,J=17.2Hz,3H),1.92(d,J=16.9Hz,4H),1.64(s ,1H),1.37(s,9H),1.27–1.09(m,6H),1.03(d,J=41.9Hz,2H),0.83(s,6H). 13CNMR(151MHz,DMSO)δ170.5,168.7,162.8,145.2,143.1,139.9,137.9,129.4,129.1,127.9,127.8,127.6,1 26.7,121.8,120.5,110.2,81.2,72.9,70.0,58.5,55.4,52.3,36.2,34.4,28.1,21.2,21.2,18.9,15.8,11.4. HRMS(ESI)m / zcalculatedforC78H88N10O14S[M]+:1432.6202;found:1432.5944. like Figures 52-53 .

[0195] Following the synthetic method of compound 9a, compound 9d, a white solid, was obtained in 32.80 g, with a yield of 72%. 1HNMR(600MHz,DMSO-d6)δ11.19(s,1H),10.80(s,1H),8.64(s,1H),8.31(s,1H),8.16(s,3H),8.06(d,J=8.1Hz,1H),7.88(d,J=7.6Hz,6H),7.76(d,J=20.1Hz,1H),7.61(s,4H),7.56(s,2H),7.40(d,J=7.7Hz,4H),7.24(s,16H),7.16(s,8H),7.05(t,J=7.5Hz,1H),7.01–6.96(m,1H),5.19(s,1H),4.69(s,1H),4.60(s,1H),4.26(s,1H),4.19(s,1H),4.12(s,4H),3.98(s,1H),3.86(s,1H),3.75(d,J=5.9Hz,1H),3.68(s,1H),3.43(s,1H),3.19(s,1H),3.11(s,1H),3.04–2.97(m,2H),2.87(d,J=33.8Hz,1H),2.74(s,2H),2.68(dd,J=14.3,11.0Hz,1H),2.57(s,1H),2.28(s,1H),1.99(s,1H),1.92(d,J=18.5Hz,2H),1.77(s,1H),1.46(s,1H),1.38(s,3H),1.26(s,1H),1.19(d,J=7.1Hz,3H),1.11(d,J=25.4Hz,3H),1.01(t,J=7.0Hz,2H),0.84(dd,J=29.7,10.3Hz,9H)。13CNMR(151MHz,DMSO)δ174.3,172.1,170.7,170.5,168.7,145.2,144.4,143.1,141.1,140.9,139.9,139.0,137.9, 136.6,135.8,133.7,129.7,129.5,129.5,129.4,129.1,129.0,128.6,128.5,128.2,127.9,127.9,127.9,127.8,126 .7,126.6,124.7,124.4,124.1,122.6,121.8,121.7,121.3,121.1,120.5,118.9,118.6,111.7,110.3,110.2,81.2,6 2.3,56.5,54.3,53.4,52.5,49.3,42.3,42.0,37.4,28.4,28.0,24.5,21.2,18.6,15.9,15.0,14.3,13.3,11.5,11.4. HRMS(ESI)m / zcalculatedforC. 83 H 90 N 10 O 14 S[M]+:1471.6360;found:1471.5630. (For example...) Figures 54-56 .

[0196] Following the synthetic method of compound 9a, 35.30 g of white solid compound 9e was obtained, with a yield of 74%. 1H1HNMR(600MHz,DMSO)δ11.18,10.78,8.58,8.29,8.18,7.96,7.89,7.85,7.70,7.66,7.64,7.62,7.58,7 .52,7.42,7.34,7.30,7.24,7.16,7.15,7.12,7.11,5.24,4.68,4.28,4.17,4.15,4.06,4.04,3.98,3.90 ,3.87,3.84,3.80,3.77,3.75,3.74,3.68,3.63,3.33,3.18,2.97,2.89,2.85,2.74,2.70,2.62,2.57,2.51,2.29,2.07,1.99,1.95,1.90,1.77,1.50,1.47,1.45,1.44,1.41,1.39,1.38,1.19,1.14,1.13,1.12,1 .10,1.09,1.08,1.08,0.91,0.90,0.85. 13CNMR(151MHz,DMSO)δ170.9,170.8,170.6,170.5,168.7,168. 0,164.0,145.2,143.1,139.9,137.9,129.4,129.1,127.9,127.8,126.7,124.7,121.8,121.3,120.5,11 8.8,118.7,111.7,110.2,81.2,73.1,72.9,70.0,60.2,58.8,58.5,55.4,54.8,54.1,53.0,49.4,42.1,40.5,36.4,36.2,36.2,34.5,34.4,31.2,28.0,28.0,24.6,21.3,21.2,18.9,15.8,15.3,14.5,11.4,11.2. HRMS(ESI)m / z calculated for C78H88N10O14S[M]+:1420.6202;found:1420.3463. (The last part is a repetition of the first and can be omitted.) Figures 57-59 .

[0197] Following the synthetic method of compound 9a, compound 9f, a white solid, was obtained in 36.80 g, with a yield of 78%. 11H NMR (600 MHz, DMSO-d6) δ 10.79 (s, 1H), 8.62 (s, 1H), 8.44 (d, J = 7.8 Hz, 1H), 8.25 (d, J = 53.8 Hz, 2H), 8.03 (dd, J = 28.3, 7.9 Hz, 2H), 7.93–7.54 (m, 6H), 7.43 (dt, J = 31.0, 6.7 Hz, 3H), 7.33–7.26 (m, 14H), 7.21–7.16 (m, 6H), 7.13–6.90 (m, 4H), 5.18 (s, 1H), 4.71 (d, J = 7.0 Hz, 1H), 4.57 (s, 1H), 4.46 (d, J = 7.2 Hz, 1H), 4.34–4.12 (m, 5H), 3.89–3.52 (m, 8H), 3.24–2.78 (m, 3H), 2.68–2.52 (m, 2H), 2.43–2.33 (m, 1H), 2.33–2.27 (m, 1H), 2.22 (d, J = 13.2 Hz, 1H), 2.10–2.00 (m, 1H), 1.88 (s, 3H), 1.74 (s, 1H), 1.40 (s, 9H), 1.24 (s, 3H), 1.07 (s, 3H), 0.90–0.82 (m, 7H), 0.79 (d, J = 7.2 Hz, 4H). 13C NMR (151 MHz, DMSO) δ 172.0, 171.4, 170.4, 170.3, 170.1, 170.1, 170.0, 169.8, 169.7, 168.6, 168.3, 162.3, 156.3, 144.7, 144.0, 143.7, 140.7, 140.6, 136.8, 128.6, 128.5, 127.6, 127.4, 127.1, 126.3, 125.9, 125.5, 125.3, 122.0, 120.1, 119.1, 118.7, 116.2, 110.9, 80.7, 72.5, 71.9, 71.5, 69.5, 66.2, 66.0, 62.9, 59.7, 58.8, 58.1, 54.9, 54.7, 51.7, 48, .1, 46.6, 42.9, 42.4, 38.0, 37.7, 35.8, 35.2, 34.0, 30.8, 27.6, 27.4, 24.4, 20.8, 20.7, 20.7, 20.6, 20.5, 15.4, 14.1, 11.0, 10.1, 10.0. HRMS (ESI) m / z calculated for C84H96N10NaO18S [M + H]+: 1565.6625; found: 1565.2549. As Figures 60-62 .

[0198] Example 10: Synthesis of compounds 10a-10f, the principle of which is shown in the following formula: .

[0199] A 2500 mL round-bottom flask was fitted with a magnetic stir bar. 49.70 g of 0.03 mol of compound 9a was added to the flask, followed by an excess of 50 wt% diethylamine in dichloromethane. The mixture was reacted at 23 °C for 2 h. After the reaction was complete, the solvent was removed under reduced pressure to obtain the crude product. The crude product was then eluted using a 200–300 mesh silica gel column with a methanol / dichloromethane volume ratio of 2%–4% to give 34.80 g of a white solid, compound 10a, in 81% yield.

[0200] 1HNMR(600MHz,DMSO-d6)δ11.12(s,1H),9.27(s,1H),8.17(s,1H),7.96(s,2H),7.90(s,1H),7.69(s,1H),7.44(s,1H),7.27(s,1H),7 .12(s,3H),6.95(s,1H),5.17(s,1H),4.65(s,1H),4.25(s,1H),4.05(s,2H),3.95(s,1H),3.73(d,J=92.7Hz,3H),3.18(d,J=5.8Hz,3 H),2.90(s,6H),2.84–2.78(m,1H),2.74(s,6H),2.60(s,1H),2.36(d,J=9.1Hz,1H),2.19(s,1H),2.09(d,J=6.8Hz,1H),1.96(d,J=2 3.4Hz,3H),1.72(s,1H),1.45(s,1H),1.34(s,1H),1.25(s,5H),1.03(d,J=82.9Hz,2H),0.87(s,5H),0.74(s,2H),0.54–0.36(m,2H). 13CNMR(151MHz,DMSO)δ173.1,171.3,171.0,171.0,170.9,170.9,170.6,17 0.3,168.3,137.0,127.6,125.0,122.6,121.0,119.0,116.4,111.5,72.8,6 2.5,59.6,58.4,54.0,53.9,53.3,51.1,49.5,42.0,41.9,40.6,39.1,38.7, 36.3,35.5,33.8,30.4,25.8,25.3,21.5,16.2,15.7,15.4,11.5,11.5,11.2. HRMS(ESI)m / zcalculatedforC44H62N10O11S[M+Na]+:965.4320,found:965.3335. like Figures 63-65 .

[0201] Following the synthetic method of compound 10a, 22.32 g of white solid compound 10b was obtained, with a yield of 84%. 1HNMR (600MHz, DMSO) δ10.80,8.72,8.64,8.28,8.21,8.03,7.98,7.91,7.90,7.80,7.59,7.58,7.33,7.31,7.27,7.21,7 .19,7.18,7.17,7.14,7.07,7.05,7.04,6.98,6.97,6.96,5.22,4.71,4.70,4.60,4.54,4.41,4.38,4.37,4.36,4.28,4 .27,4.22,4.21,4.05,4.03,3.74,3.70,3.57,3.53,3.45,3.36,3.17,3.14,3.03,2.99,2.89,2.73,2.62,2.61,2.59,2.59,2.58,2.56,2.55,2.54,2.51,2.32,2.31,2.28,2.08,2.07,2.01,2.00,1.93,1.87,1.77,1.76,1.56,1.40,1.25,1. 24,1.18,1.14,1.09,1.03,0.85,0.80,0.72,0.71,0.70,0.68,0.67. 13CNMR(151MHz,DMSO)δ174.8,172.4,172.3,170. 7,170.6,170.5,170.2,169.0,168.8,145.2,145.2,136.6,129.1,129.0,127.9,127.8,126.8,126.8,124.1,121.3,118 .8,118.6,111.7,110.3,81.2,73.1,70.0,60.2,59.8,57.2,53.5,53.4,52.5,52.4,48.5,48.2,42.6,42.0,38.3,37.4,37.2,34.5,28.5,28.1,27.9,24.7,24.5,23.7,21.3,19.2,18.9,16.2,15.8,15.7,15.0,14.6,13.3,12.0,11.5,11.4. HRMS(ESI) m / z calculated for C67H86N10O13S[M]+: 1279.6097; found: 1279.1031. (For example...) Figures 66-68 .

[0202] Following the synthetic method of compound 10a, 21.41 g of white solid compound 10c was obtained, with a yield of 83%. 1HNMR(600MHz,DMSO-d6)δ11.22(s,1H),8.65(d,J=54.6Hz,2H),8.47(s,1H),8.34(s,1H),8.23(s,4H),8.04(s,1H),7.96(s,2H),7.91 (d,J=10.2Hz,1H),7.62(s,1H),7.48(d,J=24.3Hz,2H),7.25(d,J=8.2Hz,1H),7.06(dt,J=60.8,7.5Hz,2H),5.35(d,J=17.4Hz,1H),4 .92(s,1H),4.52–4.35(m,2H),4.25(dt,J=60.0,8.1Hz,2H),4.07(s,2H),3.85(s,1H),3.63(d,J=10.1Hz,7H),3.15(s,5H),2.90(s,4 H),2.74(s,4H),2.39(s,1H),2.01(s,3H),1.35(d,J=10.2Hz,2H),1.18(d,J=9.5Hz,3H),0.90(d,J=6.7Hz,2H),0.83(d,J=3.8Hz,3H). 13CNMR(151MHz,DMSO)δ171.8,171.4,170.2,169.9,169.6,168.2,167.5,162.4,136.5,127 .4,124.3,122.2,120.1,118.6,116.0,111.1,72.3,71.2,62.8,61.9,59.8,54.9,53.9,52.8 ,52.5,52.6,50.5,48.5,42.3,41.3,39.9,39.8,39.38,39.2,39.1,38.3,35.7,34.8,34.4,34.2,33.5,31.2,31.1,30.6,29.828.9,25.2,22.0,20.9,20.7,20.5,14.8,13.9,11.2,11.1. HRMS(ESI)m / z calculated for C41H54N10O11S[M]+:891.3694;found:891.3476. For example Figures 69-71 .

[0203] Following the synthetic method of compound 10a, 20.40 g of white solid compound 10d was obtained, with a yield of 84%. 1HNMR(600MHz,DMSO-d6)δ10.80(s,1H),8.57(d,J=155.1Hz,1H),8.35–8.16(m,2H),8.08 (d,J=8.2Hz,1H),7.92(d,J=12.6Hz,1H),7.81(s,1H),7.75–7.57(m,1H),7.48(dd,J=54 .2,8.1Hz,1H),7.38–7.03(m,20H),6.97(d,J=7.6Hz,3H),5.05(d,J=160.0Hz,1H),4.69 (s,1H),4.60–4.54(m,1H),4.38(d,J=90.3Hz,1H),4.00(d,J=6.8Hz,1H),3.78(d,J=62.5 Hz,2H),3.67(s,1H),3.45(s,1H),3.21(s,1H),3.10(s,2H),3.02(t,J=9.0Hz,2H),2.86 (d,J=9.0Hz,1H),2.81–2.73(m,1H),2.70(s,1H),2.55(s,1H),2.47–2.39(m,1H),2.30( s,1H),1.95(s,1H),1.72(d,J=67.9Hz,3H),1.55–1.45(m,1H),1.36(d,J=20.3Hz,2H),1 .26(s,1H),1.22–1.12(m,4H),1.09(s,2H),1.02(s,3H),0.89–0.80(m,8H),0.69(s,1H). 13CNMR(151MHz,DMSO)δ174.4,172.1,170.7,168.7,145.1,139.1,136.6,1 29.7,129.6,129.5,129.5,129.0,128.6,128.5,128.5,127.9,126.8,126.5 ,124.1,121.3,118.9,118.6,111.7,110.3,56.5,53.4,52.6,42.3,42.0,40.9,37.4,28.4,28.1,24.5,15.9,15.7,15.0,14.5,13.3,13.1,11.5,11.4. HRMS(ESI)m / z calculated for C68H80N10O12S[M]+:1249.5678;found:1249.5056. For example Figures 72-74 .

[0204] Following the synthetic method of compound 10a, 23.43 g of white solid compound 10e was obtained, with a yield of 85%.1 HNMR(600MHz,DMSO-d6)δ11.22(s,1H),8.64(s,1H),8.48(s,1H),8.32(d,J=8. 1Hz,1H),8.08(d,J=7.5Hz,1H),8.02(d,J=8.9Hz,1H),7.91(d,J=6.0Hz,1H),7. 75(d,J=8.0Hz,1H),7.56(d,J=7.8Hz,1H),7.23(s,8H),7.22–7.14(m,8H),7.1 4–7.08(m,2H),7.01(t,J=7.5Hz,1H),5.19(s,1H),4.66(t,J=10.2Hz,2H),4.34 –4.24(m,1H),4.20(d,J=8.1Hz,1H),4.08–4.03(m,1H),3.99(d,J=8.6Hz,1H), 3.80–3.76(m,1H),3.69–3.59(m,3H),3.44–3.36(m,2H),2.90(s,2H),2.72(d,J =53.6Hz,2H),2.55(s,1H),2.33(d,J=76.6Hz,2H),2.07(s,2H),1.95(s,6H),1 .42–1.34(m,9H),1.24(s,2H),1.19(s,3H),1.10(t,J=7.2Hz,2H),0.86(s,7H). 13CNMR(151MHz,DMSO)δ172.7,172.3,170.9,170.5,170.5,170.4,170.1,1 68.7,145.2,137.7,129.0,127.9,126.7,125.3,122.3,119.5,117.4,111. 0,81.2,72.9,70.0,59.1,58.5,55.4,53.7,51.8,51.7,48.3,43.9,42.3,40.6,40.4,36.4,36.3,34.4,28.2,28.1,27.7,25.0,21.2,18.9,15.8,11.4. HRMS(ESI)m / z calculated for C63H78N10O12S[M]+:1186.5521;found:1186.5286. (The last part is a repetition of the first and can be omitted.) Figures 75-77 .

[0205] Following the synthetic method of compound 10a, 22.43 g of white solid compound 10f was obtained, with a yield of 83%. 1HNMR(600MHz, DMSO-d6) δ 11.94 (s, 1H), 8.58 (d, J = 51.4 Hz, 2H), 8.45 (s, 1H), 8.22 (d, J = 20.3 Hz, 1H), 8.03 (s, 1H), 7.90 (d, J = 31.0 Hz, 2H), 7.79 (s, 1H), 7.40 (s, 1H), 7.22 (d, J = 7.6 Hz, 7H), 7.21–7.06 (m, 12H), 5.17 (s, 1H), 4.99–4.79 (m, 2H), 4.67 (s, 2H), 4.37 (s, 1H), 4.20 (t, J = 8.0 Hz, 2H), 4.06 (s, 1H), 3.86 (s, 3H), 3.76 (d, J = 29.1 Hz, 4H), 3.67 (s, 3H), 2.96 (s, 1H), 2.58 (s, 2H), 2.27 (s, 1H), 2.01 (s, 2H), 1.96 (d, J = 19.9 Hz, 6H), 1.92 (s, 1H), 1.90 (s, 1H), 1.85 (d, J = 3.4 Hz, 1H), 1.48 (d, J = 13.4 Hz, 1H), 1.24 (s, 6H), 0.85 (d, J = 6.9 Hz, 9H). 13C NMR(151 MHz, DMSO) δ 174.5, 174.4, 172.3, 172.2, 171.2, 170.6, 170.5, 170.2, 169.8, 169.6, 169.6, 169.1, 168.8, 162.8, 145.2, 145.0, 144.8, 144.7, 129.6, 128.0, 127.2, 126.9, 124.2, 124.1, 121.3, 118.9, 118.6, 111.7, 110.1, 110.0, 82.2, 81.1, 80.8, 72.7, 70.0, 69.9, 66.2, 66.2, 60.5, 58.6, 57.1, 55.3, 53.2, 52.2, 51.7, 51.6, 48.9, 48.6, 48.3, 46.9, 42.5, 42.0, 38.5, 37.4, 36.2, 34.5, 34.3, 31.2, 30.5, 28.5, 28.4, 28.1, 27.9, 26.0, 26.0, 24.8, 21.3, 21.2, 21.2, 21.0, 18.7, 18.7, 18.5, 15.8, 15.3, 11.8, 11.5, 11.5, 11.2. HRMS(ESI) m / z calculated for C69H86N10O16S [M + H]+: 1343.5944; found: 1343.1350. As Figures 78-80 .

[0206] Example 11: Synthesis of compounds 11a-11f, the principle of which is shown in the following formula: .

[0207] A 2500 mL round-bottom flask was fitted with a magnetic stir bar. 28.70 g of 0.02 mol of compound 10a was added to the flask, followed by 200 mL of trifluoroacetic acid. The mixture was stirred at room temperature for 0.5 h, and the trifluoroacetic acid was removed under reduced pressure to obtain the crude product. The crude product was not further purified and was used directly in the next synthesis. The crude product was dissolved in dry N,N-dimethylformamide and stirred at 0°C for 5 min. Then, 6.90 g of 2 equiv diisopropylethylamine and 15.20 g of 2 equiv 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate were added. The mixture was heated to 23°C and reacted for 12 h. After the reaction was complete, N,N-dimethylformamide was removed under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography with gradient elution of 4%–6% methanol / dichloromethane by volume to obtain a purified white solid compound 11a, 13.56 g, with a yield of 61%. 1HNMR(600MHz,DMSO-d6)δ11.12(s,1H),9.27(s,1H),8.17(s,1H),7.96(s,2H),7.90(s,1H),7.69(s,1H),7.44(s,1H),7.27(s,1H),7 .12(s,3H),6.95(s,1H),5.17(s,1H),4.65(s,1H),4.25(s,1H),4.05(s,2H),3.95(s,1H),3.73(d,J=92.7Hz,3H),3.18(d,J=5.8Hz,3 H),2.90(s,6H),2.84–2.78(m,1H),2.74(s,6H),2.60(s,1H),2.36(d,J=9.1Hz,1H),2.19(s,1H),2.09(d,J=6.8Hz,1H),1.96(d,J=2 3.4Hz,3H),1.72(s,1H),1.45(s,1H),1.34(s,1H),1.25(s,5H),1.03(d,J=82.9Hz,2H),0.87(s,5H),0.74(s,2H),0.54–0.36(m,2H). 13CNMR(151MHz,DMSO)δ173.1,171.3,171.0,171.0,170.9,170.9,170.6,17 0.3,168.3,137.0,127.6,125.0,122.6,121.0,119.0,116.4,111.5,72.8,6 2.5,59.6,58.4,54.0,53.9,53.3,51.1,49.5,42.0,41.9,40.6,39.1,38.7, 36.3,35.5,33.8,30.4,25.8,25.3,21.5,16.2,15.7,15.4,11.5,11.5,11.2. HRMS(ESI)m / zcalculatedforC 44 H 62 N 10 O 11 S[M+Na]+:965.4320,found:965.3335. (For example...) Figures 81-83 .

[0208] Following the synthetic method of compound 11a, 6.25 g of white solid compound 11b was obtained, with a yield of 62%. 1HNMR(600MHz,DMSO-d6)δ11.24(s,1H),8.56(d,J=7.4Hz,1H),8.48(s,1H),8.36(s,1H),8.20(s,1H),8.11(d,J=9.1Hz,1H),8.06(d,J=9.9Hz,1H),8.01(d,J=7.2Hz,1H),7.88(d,J=10.1Hz,1H),7.72(s,1H),7.61(d,J=8.0Hz,1H),7.46(d,J=12.0Hz,4H),7.32(s,2H),7.25(d,J=8.2Hz,1H),7.20(d,J=10.9Hz,1H),7.11(t,J=7.5Hz,1H),7.01(t,J=7.5Hz,1H),5.33(s,1H),5.19(d,J=6.2Hz,1H),5.00–4.94(m,1H),4.70(s,1H),4.48(t,J=13.0Hz,1H),4.39–4.28(m,3H),4.21(t,J=8.9Hz,1H),3.89–3.80(m,2H),3.76(s,1H),3.61(s,1H),3.41(s,1H),3.26–3.16(m,2H),3.12–3.06(m,1H),2.93(dd,J=15.3,8.6Hz,4H),2.81(d,J=10.4Hz,1H),2.62(s,1H),2.39(s,1H),2.27(d,J=41.9Hz,2H),1.99(d,J=13.3Hz,3H),1.82(s,1H),1.58(s,1H),1.46(s,3H),1.34(d,J=7.2Hz,3H),1.24(s,3H),1.17(t,J=7.2Hz,6H),0.89(d,J=6.7Hz,3H),0.82(d,J=6.9Hz,6H)。13CNMR(151MHz,DMSO)δ173.1,171.3,171.0,171.0,170.9,170.9,170.6,170.3,168.3,137.0,127.6,125.0,122.6,121.0,119.0,116.4,111.5,72.8,62.5,59.6,58.4,54.0,53.9,53.3,51.1,49.5,42.0,41.9,40.6,39.1,38.7,36.3,35.5,33.8,30.4,25.8,25.3,21.5,16.2,15.7,15.4,11.5,11.5,11.2。HRMS(ESI)m / z calculated for C. 40 H 54 N 10 O 11 S[M]+:882.3694;found:1112.4630. (For example...) Figures 84-86 .

[0209] Following the synthetic method of compound 11a, 6.25 g of white solid compound 11c was obtained, with a yield of 62%. 1HNMR(600MHz, DMSO-d6) δ 11.94 (s, 1H), 11.68 (s, 1H), 8.42 (s, 1H), 8.36 (s, 1H), 8.27 (s, 1H), 8.06 (s, 1H), 7.95 (d, J = 9.0 Hz, 1H), 7.91–7.88 (m, 1H), 7.80 (s, 1H), 7.58 (s, 1H), 7.47 (dd, J = 14.0, 6.5 Hz, 1H), 7.33 (s, 1H), 7.18 (d, J = 7.6 Hz, 1H), 5.34 (d, J = 3.8 Hz, 1H), 5.07 (t, J = 11.2 Hz, 1H), 5.04 (s, 1H), 4.95 (s, 1H), 4.86 (s, 1H), 4.46 (dd, J = 18.1, 9.2 Hz, 1H), 4.31 (d, J = 44.2 Hz, 1H), 4.18 (s, 1H), 4.12–4.03 (m, 1H), 4.00 (s, 1H), 3.86 (d, J = 13.4 Hz, 1H), 3.60 (s, 1H), 3.54–3.39 (m, 2H), 3.24 (d, J = 15.4 Hz, 1H), 3.17 (s, 1H), 3.04 (s, 1H), 2.92 (s, 3H), 2.74 (s, 1H), 2.38 (d, J = 18.0 Hz, 1H), 2.06 (s, 3H), 1.91 (s, 1H), 1.58 (s, 1H), 1.48 (s, 2H), 1.34 (s, 3H), 1.24 (s, 4H), 1.16 (s, 3H), 1.05 (s, 1H), 0.90 (s, 2H), 0.83 (qd, J = 6.3, 5.4, 3.3 Hz, 7H). 13C NMR(151 MHz, DMSO) δ 172.0, 171.9, 171.1, 170.7, 170.2, 169.9, 169.8, 169.7, 169.6, 168.2, 167.5, 136.5, 127.0, 124.3, 122.2, 120.1, 118.7, 116.1, 111.2, 72.3, 71.2, 62.9, 61.9, 59.0, 54.9, 53.0, 52.9, 52.5, 50.5, 42.3, 41.4, 38.3, 34.9, 34.4, 34.3, 33.6, 30.6, 29.0, 25.2, 21.0, 20.7, 20.6, 14.8, 11.2, 11.0, 10.6. HRMS(ESI) m / z calculated for C 44 H 62 N 10 O 12 S [M+Na]+: 975.4269; found: 975.3335. As Figures 87-89 .

[0210] Following the synthetic method of compound 11a, 6.98 g of white solid compound 11d was obtained, with a yield of 67%. 1 HNMR(600MHz,DMSO-d6)δ11.22(s,1H),8.65(d,J=54.6Hz,2H),8.47(s,1H),8.34(s,1H),8.23(s,4H),8.04(s,1H),7.96(s,2H),7.91 (d,J=10.2Hz,1H),7.62(s,1H),7.48(d,J=24.3Hz,2H),7.25(d,J=8.2Hz,1H),7.06(dt,J=60.8,7.5Hz,2H),5.35(d,J=17.4Hz,1H),4 .92(s,1H),4.52–4.35(m,2H),4.25(dt,J=60.0,8.1Hz,2H),4.07(s,2H),3.85(s,1H),3.63(d,J=10.1Hz,7H),3.15(s,5H),2.90(s,4 H),2.74(s,4H),2.39(s,1H),2.01(s,3H),1.35(d,J=10.2Hz,2H),1.18(d,J=9.5Hz,3H),0.90(d,J=6.7Hz,2H),0.83(d,J=3.8Hz,3H). 13CNMR(151MHz,DMSO)δ171.8,171.4,170.2,169.9,169.6,168.2,167.5,162.4,136.5,127 .4,124.3,122.2,120.1,118.6,116.0,111.1,72.3,71.2,62.8,61.9,59.8,54.9,53.9,52.8 ,52.5,52.6,50.5,48.5,42.3,41.3,39.9,39.8,39.38,39.2,39.1,38.3,35.7,34.8,34.4,34.2,33.5,31.2,31.1,30.6,29.828.9,25.2,22.0,20.9,20.7,20.5,14.8,13.9,11.2,11.1. HRMS(ESI)m / z calculated for C41H54N10O11S[M]+:891.3694;found:891.3476. For example Figures 90-92 .

[0211] Following the synthetic method of compound 11a, 6.69 g of white solid compound 11e was obtained, with a yield of 63%.1 HNMR(600MHz,DMSO-d6)δ11.27(s,1H),8.57(s,1H),8.45(s,1H),8.38(d,J=16.8 Hz,2H),8.16(s,2H),7.98(d,J=22.4Hz,2H),7.89(s,1H),7.72(s,1H),7.61(s,1H ),7.52(s,1H),7.32(s,2H),7.24(d,J=40.4Hz,5H),7.13(t,J=7.9Hz,2H),7.05(s ,2H),5.29(s,1H),5.04(d,J=39.8Hz,2H),4.52(d,J=11.5Hz,1H),4.37(s,2H),4. 23(s,2H),4.09(s,2H),3.96(d,J=23.3Hz,2H),3.64(s,1H),3.49(s,1H),2.98(s, 2H),2.94(s,4H),2.90–2.85(m,2H),2.74(s,1H),2.62(s,1H),2.39(s,2H),2.18( s,1H),2.09(s,1H),2.00(d,J=10.4Hz,3H),1.88(s,1H),1.60(s,1H),1.46(dd,J= 16.0,8.3Hz,2H),1.39–1.33(m,4H),1.24(s,3H),1.15(s,7H),0.84–0.83(m,3H). 13CNMR(151MHz,DMSO)δ173.4,172.5,172.4,171.0,170.9,170.7,170.5,170.3,169.5,137.7,127.4,125.4,122.5,119.7,119.0,117.4,11 1.4,73.1,59.8,58.5,53.2,52.2,51.9,49.3,48.2,43.9,37.4,37.2, 36.6,36.4,34.6,34.5,28.5,25.0,21.3,21.2,19.0,16.0,15.8,11.4. HRMS(ESI) m / z calculated for C44H56N10O11S[M+Na]+: 953.3851; found: 953.3481. (For example...) Figures 93-95 .

[0212] Following the synthetic method of compound 11a, 6.08 g of white solid compound 11f was obtained, with a yield of 61%. 1HNMR(600MHz,DMSO-d6)δ11.64(s,1H),8.86(s,1H),8.54(s,1H),8.30(s,1H),8.26(s,2H),7.87(dd,J=16.3,9.6Hz,2H),7.81(d,J=9.2Hz,1H),7.66(d,J=8.1Hz,1H),7.42(d,J=8.3Hz,2H),7.20(t,J=7.6Hz,2H),7.10(t,J=7.6Hz,1H),5.35(s,2H),5.22(s,1H),5.02(s,1H),4.91(s,1H),4.86(d,J=6.8Hz,1H),4.78(s,1H),4.33–4.26(m,3H),4.09(s,3H),3.89(d,J=7.0Hz,1H),3.71(s,1H),3.50(s,1H),3.48(s,1H),3.44(s,1H),3.41(s,1H),3.18(s,1H),3.11(s,2H),2.96(s,2H),2.84(d,J=7.3Hz,1H),2.75(d,J=14.0Hz,1H),2.62(s,1H),2.56(ddd,J=10.6,7.0,3.6Hz,2H),2.39(s,1H),2.26(dd,J=13.8,7.0Hz,1H),2.12(s,2H),2.03(s,3H),1.99(s,3H),1.96(s,3H),1.90(d,J=16.0Hz,1H),1.56(s,3H),1.46(s,1H),1.30(d,J=7.1Hz,1H),1.24(s,6H),1.12(d,J=7.3Hz,2H),0.93(d,J=7.0Hz,3H),0.84(d,J=7.4Hz,3H),0.81(s,3H)。13CNMR(151MHz,DMSO)δ172.1,171.8,171.2,170.6,170.4,170.3,170.2,169.8,169.8,1 69.6,169.5,168.0,167.0,162.4,147.1,137.3,133.0,125.1,121.9,114.5,111.1,105.1 ,72.4,71.1,62.9,61.9,59.8,59.2,52.9,52.7,52.4,50.8,50.0,42.4,41.2,35.8,34.8 ,34.7,34.6,33.0,30.8,29.1,25.3,21.0,20.9,20.8,20.7,20.6,14.8,14.1,11.1,10.8. HRMS(ESI)m / zcalculatedforC46H62N10O15S[M+H]+:1026.4117;found:1026.2549. like. Figures 96-98 .

[0213] Example 12: Synthesis of compounds 12a-12f, the principle of which is shown in the following formula: .

[0214] Take a 2500 mL round-bottom flask and place a magnetic stir bar inside. Add 5.60 g of 0.005 mol of compound 11a to the flask, followed by 300 mL of a mixed solution of isopropanol and ethanol (volume ratio 2:1). Stir in an ice-water bath for 10 min. Add dropwise a mixed solution containing 0.9 equiv m-chloroperoxybenzoic acid to bring the volume of the mixed solution to 50 mL. React at 0°C for 30 min. After the reaction is complete, remove the mixture of isopropanol and ethanol under reduced pressure to obtain the crude product. Analyze the crude product using a 200–300 mesh silica gel column with a methanol / dichloromethane elution system of 5%–6% (v / v) to obtain 3.90 g of a white solid, compound 12a, in 70% yield.

[0215] 1HNMR(500MHz,DMSO-d6)δ11.26(s,1H),8.59(d,J=7.3Hz,1H),8.50(s,1H),8.38(s, 1H),8.23(s,1H),8.11–8.02(m,2H),7.90(d,J=10.3Hz,1H),7.74(s,1H),7.61(d,J= 8.0Hz,1H),7.52–7.44(m,3H),7.35(s,1H),7.25(d,J=8.1Hz,1H),7.12(t,J=7.5Hz, 1H),7.01(t,J=7.5Hz,1H),5.33(s,1H),5.21(d,J=3.4Hz,1H),4.96(d,J=6.3Hz,1H) ,4.71(s,1H),4.49(s,1H),4.29(s,3H),4.21(t,J=9.0Hz,1H),3.83(d,J=12.0Hz,2H ),3.76(s,1H),3.60(s,1H),3.22(s,1H),3.09(dd,J=14.6,6.3Hz,1H),2.91(q,J=7. 2Hz,4H),2.81(d,J=8.8Hz,1H),2.65(s,1H),2.24(s,1H),1.98(s,2H),1.83(t,J=12 .1Hz,1H),1.48(s,3H),1.37–1.22(m,8H),1.17(t,J=7.2Hz,5H),0.93–0.79(m,9H). 13CNMR(151MHz,DMSO)δ172.9,171.3,171.1,170.6,167.8,166.9,156.8,144.6,140.6,136.7,136.4,129.6,128.1,122.8,120.5,116 .2,69.1,67.7,62.0,59.6,58.6,55.5,53.3,51.5,49.1,38.8,38.0,35.6,34.1,33.8,28.6,25.8,25.4,19.0,16.2,15.4,11.5,11.3. HRMS(ESI)m / zcalculatedforC 42 H 60 N 10 O 10 S[M]+:891.4215,found:891.3476. (For example...) Figures 99-101 .

[0216] Following the synthetic method of compound 12a, compound 12b, a white solid, was obtained in 3.00 g, with a yield of 73%. 11H NMR (600 MHz, DMSO-d6) δ 8.53 (s, 1H), 8.44 (s, 1H), 8.34 (s, 1H), 8.21 (d, J = 7.0 Hz, 1H), 8.03 (d, J = 9.8 Hz, 2H), 7.97–7.87 (m, 2H), 7.80 (d, J = 8.2 Hz, 1H), 7.72 (s, 1H), 7.50–7.39 (m, 4H), 7.32 (d, J = 11.4 Hz, 2H), 7.17 (t, J = 7.6 Hz, 1H), 5.18 (s, 1H), 5.10–4.93 (m, 2H), 4.68 (d, J = 26.1 Hz, 1H), 4.43 (dd, J = 18.2, 9.2 Hz, 1H), 4.36 (t, J = 5.2 Hz, 2H), 4.31–4.26 (m, 1H), 4.20–4.07 (m, 2H), 3.89 (t, J = 13.1 Hz, 1H), 3.79 (d, J = 11.1 Hz, 1H), 3.72 (d, J = 11.2 Hz, 1H), 3.60 (s, 1H), 3.51–3.38 (m, 3H), 3.29–3.19 (m, 1H), 3.17 (d, J = 4.5 Hz, 2H), 3.07–2.87 (m, 2H), 2.24–2.17 (m, 1H), 1.93 (s, 1H), 1.85–1.77 (m, 1H), 1.57 (s, 1H), 1.48 (s, 2H), 1.40–1.29 (m, 3H), 1.25 (d, J = 8.6 Hz, 2H), 1.16 (d, J = 7.2 Hz, 2H), 1.06 (t, J = 6.9 Hz, 3H), 0.90 (d, J = 6.8 Hz, 2H), 0.82 (d, J = 7.2 Hz, 6H). 13C NMR (151 MHz, DMSO) δ 172.9, 171.5, 171.3, 171.1, 170.6, 168.1, 143.2, 136.7, 136.4, 132.8, 130.0, 129.8, 129.6, 129.3, 128.1, 127.5, 125.7, 124.0, 123.3, 122.0, 120.6, 116.4, 113.4, 69.1, 62.3, 58.6, 56.1, 53.4, 50.9, 49.8, 48.5, 38.4, 36.2, 35.6, 25.8, 25.4, 19.0, 16.2, 15.4, 11.5, 11.3. HRMS (ESI) m / z calculated for C42H60N10O11S [M+Na]+: 935.4416, found: 935.3228. As Figures 102-104 .

[0217] Following the synthetic method of compound 12a, compound 12c, a white solid, was obtained in 3.40 g, with a yield of 74%. 1 HNMR(600MHz,DMSO-d6)δ11.23(s,1H),8.59–8.45(m,2H),8.35(s,1H),8.20(s,1H),8. 09(dd,J=26.0,9.4Hz,2H),8.01(d,J=7.3Hz,1H),7.88(s,1H),7.61(d,J=8.0Hz,1H),7 .47(s,2H),7.25(d,J=8.2Hz,1H),7.11(t,J=7.5Hz,1H),7.02(s,1H),5.18(s,1H),4.9 6(d,J=12.9Hz,1H),4.69(s,1H),4.48(s,1H),4.38(s,3H),4.21(s,1H),3.82(d,J=11.8 Hz,2H),3.74(d,J=11.1Hz,1H),3.61(s,2H),3.43–3.41(m,2H),3.24(d,J=14.2Hz,2H) ,3.11–3.07(m,1H),2.95(s,2H),2.81(q,J=7.3Hz,2H),2.40(s,7H),2.26(s,6H),1.95 (s,2H),1.82(d,J=12.1Hz,1H),1.69(d,J=3.8Hz,3H),1.57(s,1H),1.47(s,2H),1.34( d,J=7.1Hz,3H),1.24(s,2H),1.13(d,J=7.2Hz,2H),0.89(s,3H),0.82(d,J=6.7Hz,6H). 13CNMR(151MHz,DMSO)δ172.9,171.2,171.2,171.1,171.0,170.9,170.9,170.8,170.6,168 .4,137.0,130.1,129.6,128.1,127.6,124.9,122.6,121.0,119.0,116.5,111.5,69.1,62.3 ,59.6,58.3,56.2,56.1,54.0,53.8,51.2,49.6,42.1,42.0,38.9,38.5,36.2,35.5,34.3,30 .6,29.5,29.3,29.0,27.0,25.8,25.3,24.0,22.5,16.3,15.6,15.3,14.4,11.9,11.4,11.3. HRMS(ESI)m / zcalculated for C 40 H54 N 10 O 11 S[M+Na]+:854.3589,found:854.1330. (Example) Figures 105-107 .

[0218] Following the synthetic method of compound 12a, compound 12d, a white solid, was obtained in 3.42 g, with a yield of 73%. 1HNMR(600MHz,DMSO-d6)δ10.81(s,1H),8.26–8.19(m,1H),8.16(s,1H),7. 96(d,J=14.5Hz,1H),7.88(s,2H),7.72(d,J=15.2Hz,1H),7.63(d,J=26.7 Hz,4H),7.41(s,3H),7.30(d,J=8.2Hz,5H),7.24(s,4H),7.19(s,2H),7.0 5(s,1H),6.97(s,1H),4.58(d,J=21.3Hz,1H),4.27(s,1H),4.19(s,1H),4 .12(s,2H),4.04(s,2H),3.75(s,1H),3.44(s,1H),3.33(s,4H),3.18(d,J =19.0Hz,1H),3.09(s,1H),2.99(d,J=9.4Hz,2H),2.74(s,1H),1.83(d,J= 75.9Hz,1H),1.48(s,1H),1.41(s,1H),1.26(d,J=25.6Hz,1H),1.18(s,2H ),1.13(s,2H),1.07(q,J=7.5Hz,1H),1.01(s,2H),0.85(d,J=43.2Hz,6H). 13CNMR(151MHz,DMSO)δ173.1,172.8,170.8,170.7,168.6,156.2,145.2,144.7,144.3,144.2,141.1, 138.6,136.5,129.7,129.7,129.6,129.5,128.6,128.5,128.3,127.9,127.6,127.5,126.6,125.8,125 .7,124.9,124.1,121.4,121.3,120.5,119.6,118.8,118.7,111.7,110.3,110.1,66.1,60.2,56.6,53.9,52.5,47.1,42.4,42.0,37.9,37.5,28.9,28.1,24.5,21.2,15.7,15.0,14.6,14.5,13.3,13.2,11.4. HRMS(ESI) m / z calculated for C42H54N10O10S[M+Na]+:923.3745,found:923.3267. (The last part is a separate, unrelated statement and can be omitted.) Figures 108-110 .

[0219] Following the synthetic method of compound 12a, 3.39 g of white solid compound 12e was obtained, with a yield of 72%.1 HNMR (600 MHz, DMSO-d6) δ 11.22 (s, 1H), 8.58–8.47 (m, 2H), 8.35 (s, 1H), 8.19 (s, 1H), 8.10 (d, J = 9.5 Hz, 1H), 8.06 (d, J = 10.0 Hz, 1H), 8.01 (d, J = 7.3 Hz, 1H), 7.87 (d, J = 10.2 Hz, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.46 (d, J = 18.7 Hz, 1H), 7.25 (d, J = 8.1 Hz, 1H), 7.11 (t, J = 7.6 Hz, 1H), 7.01 (t, J = 7.5 Hz, 1H), 5.17 (d, J = 3.4 Hz, 1H), 4.97 (dt, J = 13.5, 7.0 Hz, 1H), 4.71 (d, J = 3.9 Hz, 1H), 4.52–4.48 (m, 1H), 4.38 (s, 1H), 4.37–4.29 (m, 2H), 4.22 (d, J = 9.9 Hz, 1H), 3.82 (d, J = 7.9 Hz, 2H), 3.74 (d, J = 11.1 Hz, 1H), 3.64–3.59 (m, 1H), 3.40 (dd, J = 16.0, 3.6 Hz, 2H), 3.23 (s, 1H), 3.13–3.07 (m, 1H), 2.98–2.88 (m, 3H), 2.81 (d, J = 10.9 Hz, 1H), 2.22 (s, 1H), 1.96 (d, J = 7.9 Hz, 1H), 1.83 (t, J = 14.0 Hz, 1H), 1.58 (s, 1H), 1.47 (s, 2H), 1.34 (d, J = 7.2 Hz, 3H), 1.24 (s, 3H), 1.16 (d, J = 14.5 Hz, 4H), 0.90 (s, 3H), 0.82 (d, J = 6.9 Hz, 7H). 13CNMR (151 MHz, DMSO) δ 173.4, 172.5, 172.4, 171.0, 170.9, 170.7, 170.5, 170.3, 169.5, 137.7, 127.4, 125.4, 122.5, 119.7, 119.0, 117.4, 111.4, 73.1, 59.8, 58.5, 53.2, 52.2, 51.9, 49.3, 48.2, 43.9, 37.4, 37.2, 36.6, 36.4, 34.6, 34.5, 28.5, 25.0, 21.3, 21.2, 19.0, 16.0, 15.8, 11.4. HRMS (ESI) m / z calculated for C38H52N10O10S [M+Na]+: 863.3589, found: 863.3621. As Figures 111-113 .

[0220] Following the synthetic method of compound 12a, compound 12f, a white solid, was obtained in 3.36 g, with a yield of 71%. 1 HNMR(600MHz,DMSO-d6)δ11.64(s,1H),8.76(s,1H),8.54(s,1H),8.39(s,1H),8.36–8.18(m,2H),7.90(dd,J=42.7,9.1Hz,2H),7.67(t,J=8.6H z,2H),7.47–7.34(m,2H),7.26–7.18(m,1H),7.10(d,J=7.5Hz,1H),5.1 8(s,2H),5.00(s,1H),4.88(s,1H),4.67(s,1H),4.37(s,2H),4.32–4.19 (m,2H),4.05(dd,J=11.4,8.4Hz,1H),3.91(d,J=9.9Hz,1H),3.82(s,1H),3.76(s,1H),3.69(s,1H),3.45(s,4H),3.24(d,J=6.8Hz,1H),3.15(t ,J=13.3Hz,1H),2.97(s,1H),2.79(s,1H),2.22(s,1H),2.11–1.95(m,1 H),1.88(s,1H),1.57(s,2H),1.45–0.99(m,3H),0.82(d,J=16.0Hz,8H). 13CNMR(151MHz,DMSO)δ171.9,171.7,171.2,171.0,170.8,170.2,169.9,167.9,167.0,137.4,132.4,127.2,123.1,121.5,119.4,112.3,111.0 ,71.9,68.6,63.5,62.1,59.2,59.0,55.8,54.0,52.6,50.9,50.0,48.6 ,42.4,41.2,40.1,37.8,37.3,34.6,33.4,29.2,25.2,14.8,11.5,10.7. HRMS(ESI)m / z calculated for C44H60N10O14S[M]+:981.4011;found:981.4199. (For example...) Figures 114-116 .

[0221] Cell viability experiments were performed on the 11a~11f and 12a~12f cells prepared above. The specific procedures are as follows: Cell viability assay procedure: Human cancer cell lines Hela and SKOV3 were purchased from ProCell (Wuhan, China) and cultured under standard conditions: 37°C, 5% CO2 by volume in the air, and 95% humidity.

[0222] HeLa cells were cultured in Dulbecco modified Eagle medium (keygenBIO, KGL1206-500) containing L-alanylglutamine and supplemented with 10% fetal bovine serum and 1 mM sodium pyruvate.

[0223] SKOV3 cells were cultured in DMEM containing L-alanylglutamine and supplemented with 10% FBS and 1 mM sodium pyruvate. For passage, adherent cells were detached by adding TrypLEExpress (1×) (Gibco, 12604-013) and transferred to new culture flasks after dilution with PBS.

[0224] To evaluate amantinib derivatives and drug-conjugates, as well as amantinib derivative-ADCs, 2 μL of live cell suspension was seeded into 96-well plates, with 5000 vc per well for HeLa cells and 5000 vc per well for SKOV3 cells, and then cultured overnight in a humid chamber at 37°C.

[0225] Test compounds were prepared at a 10-fold starting concentration and serially diluted. Tests 11a–11f and 12a–12f were prepared at a starting concentration of 5 μg / mL using a serial dilution method, i.e., one part sample solution was mixed with one part dilution buffer, and 12 μL was added to the cells. The experiment was performed using CellTiter-Glo reagent according to the manufacturer's instructions, and cell viability was measured after 72 hours. The contents were mixed on an orbital oscillator for 2 minutes to induce cell lysis, and then incubated at room temperature for 10 minutes to stabilize the luminescence signal.

[0226] The luminescence values ​​were measured using a TECANSpark® multimode microplate reader (Tecan Group Ltd.) and normalized to the luminescence values ​​of untreated control cells.

[0227] The dose-response curves were fitted using a four-parameter function of logarithm versus slope of the response variable for the inhibitor GraphPadPrism (GraphPad Software, Inc.). Data are presented as the percentage of relative cell viability relative to the molar concentration of the compound. IC50 was calculated through multiple experiments. 50 The geometric mean of the values ​​was used. The luminescence values ​​from the carrier control group were set as 100% survival rate. The relative cell viability (%) of the treatment groups was calculated using the following formula: (Treatment group luminescence value / Control group luminescence value) × 100%. Results are as follows... Figure 1 and Figure 2As shown. As expected, based on results from both cell lines, the half-maximal inhibitory concentration (IC50) of product 8 in HeLa cells was [value missing]. 50 The concentration was 1.307 μM in HeLa cells and 5.096 μM in SKOV3 cells. The half-maximal inhibitory concentration (IC50) of HeLa cells after treatment with products 11 and 12 was [not specified]. 50 The half-maximal inhibitory concentrations (IC50) for SKOV3 cells were 6.749 μM and 1.912 μM, respectively. 50 The concentrations were 6.241 μM and 1.830 μM, respectively. Surprisingly, product 12, containing dihydroxyisoleucine and an -OAc protecting group, exhibited superior antitumor activity compared to other compounds. Notably, the introduction of the non-natural amino acid dihydroxyisoleucine significantly enhanced the cellular activity of the derivative.

[0228] Those skilled in the art will readily understand that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A cyclic peptide toxin Amanitin derivative, characterized in that, The structure is as follows: 。 2. The use of the cyclic peptide toxin Amanitin derivative of claim 1 as a toxin loading material in the preparation of antibody-drug conjugates.

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