Method for preparing cyclic peptide toxin α-amanitin and / or amaninamide, and intermediate and use thereof

Abstract

The present application belongs to the field of organic synthesis, and provided is a method for preparing cyclic peptide toxin α-Amanitin and / or Amanamide, and an intermediate and the use thereof. Provided is a complete synthesis method for cyclic peptide toxin compounds α-Amanitin and Amanamide, wherein the used raw materials and reagents are easy to purchase commercially, the intermediate is stable, the reaction conditions are mild, the operation process is simple and convenient, and the separation and purification process is good in terms of operability, is high in terms of yield, and has an important reference and practical value, which can realize gram-scale preparation of α-Amanitin and Amanamide and has good industrial prospects. The preparation method has a significant application value in the field of cyclic peptide toxin synthesis.

Description

Preparation method, intermediates and applications of cyclic peptide toxin α-Amanitin and / or Amaninamide

[0001] This application claims priority to a Chinese patent application filed with the Patent Office of China on March 7, 2024, with application number CN 202410257131.1 and invention name “Method for preparing cyclic peptide toxin α-Amanitin and / or Amaninamide, intermediates and applications”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present application relates to the technical field of organic synthesis, and in particular to a preparation method, intermediates and applications of cyclic peptide toxins α-Amanitin and / or Amaninamide. Background Art

[0003] Amanitin is a cyclic peptide isolated from highly toxic mushrooms and belongs to the amanitin family of toxins. Amanitin is a bicyclic octapeptide structure comprising nine distinct peptides: α-amanitin, β-amanitin, γ-amanitin, ε-amanitin, amanin, amaninamide, amanullin, amanullinic acid, and proamanullin. α-Amanitin is the most abundant bicyclic octapeptide in the amanitin family, and its antibody-drug conjugates (ADCs) are used in cancer therapy. Consequently, research on its chemical properties and applications is highly sought after.

[0004] α-Amanitin is a highly potent inhibitor of eukaryotic RNA polymerase II, blocking mRNA transcription and protein synthesis while simultaneously killing both rapidly growing and quiescent cells, leading to their rapid degradation and subsequent apoptosis. Currently, α-Amanitin antibody-drug conjugates have cured pancreatic cancer in xenograft mice and are progressing toward human trials. Medical researchers require sufficient quantities of α-Amanitin to prepare their ADCs for cancer treatment. α-Amanitin is primarily isolated and extracted from naturally occurring poisonous mushrooms, and can also be synthesized by fermentation. However, the yields of natural product extraction and fermentation are low, and the market price is extremely high (approximately 12,000 CNY / mg), severely hindering the development of antibody-drug conjugates for cancer therapy. Chemical synthesis of α-Amanitin is a proven method with the potential for large-scale production. Although Heinrich Wieland et al. characterized the structure and properties of α-Amanitin and its derivatives in the 1940s, chemists have also been attempting to chemically synthesize α-Amanitin.

[0005] So far, there is still a lack of efficient and large-scale methods for synthesizing α-Amanitin and its derivatives.

[0006] Summary of the Invention

[0007] In view of this, the purpose of this application is to provide a preparation method, intermediates, and applications of the cyclic peptide toxin α-Amanitin and / or Amaninamide. The preparation method provided in this application is a total synthesis method with readily available raw materials, stable intermediates, mild reaction conditions, simple operation process, good operability of the separation and purification process, and high yield.

[0008] In order to achieve the above application objectives, this application provides the following technical solutions:

[0009] The present application provides a method for preparing cyclic peptide toxin compounds α-Amanitin and / or Amaninamide, wherein the α-Amanitin has a structure shown in Formula 1a, and the Amaninamide has a structure shown in Formula 1b:

[0010] The following steps are involved:

[0011] Step 1: subjecting compound 2 to acetylation and esterification to obtain compound 3;

[0012] Step 2: subjecting the compound 3 to a condensation reaction and a deprotection reaction to obtain compound 4;

[0013] Step 3: subjecting the compound 4 to condensation reaction and deprotection reaction to obtain compound 5;

[0014] Step 4: subjecting the compound 5 to condensation reaction and deprotection reaction to obtain compound 6;

[0015] Step 5: subjecting the compound 6 to condensation reaction and deprotection reaction to obtain compound 7;

[0016] Step 6: subjecting the compound 7 to condensation reaction and deprotection reaction to obtain compound 8;

[0017] Step 7: subjecting the compound 8 to condensation reaction and deprotection reaction to obtain compound 9;

[0018] Step 8: subjecting the compound 9 to condensation reaction and deprotection reaction to obtain compound 10;

[0019] Step 9: Compound 10 is subjected to carbon-sulfur ring closure in an iodine-mediated manner to obtain compound 11;

[0020] Step 10: subjecting the compound 11 to a deprotection reaction to obtain compound 12;

[0021] Step 11: subjecting the compound 12 to a deprotection reaction and a condensation reaction to obtain compound 13;

[0022] Step 12: subjecting the compound 13 to an oxidation reaction to obtain compound 14;

[0023] Step 13: subjecting the compound 14 to a deprotection reaction to obtain the cyclic peptide toxin compounds α-Amanitin and Amaninamide;

[0024] The structures of compounds 2 to 14 are as follows:

[0025] In the compounds 9 to 14, R is -H or -OBn.

[0026] Preferably, the preparation method comprises the following steps:

[0027] Step 1: Compound 2, pyridine, acetyl chloride and an organic solvent are mixed for acetylation reaction, and the obtained acetylated product is mixed with N,N-dimethylpyridine and di-tert-butyl dicarbonate for esterification reaction to obtain Compound 3;

[0028] Step 2: mixing the compound 3, N-Fmoc-asparagine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 4;

[0029] Step 3: mixing the compound 4, N-Fmoc-cysteine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 5;

[0030] Step 4: mixing the compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 6;

[0031] Step 5: mixing the compound 6, N-Fmoc-isoleucine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 7;

[0032] Step 6: Mixing the compound 7, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 8;

[0033] Step 7: Mixing the compound 8, compound 15, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent for condensation reaction and deprotection reaction to obtain the compound 9; the compound 15 is N-Fmoc-6-benzyloxytryptophan or N-Fmoc-tryptophan;

[0034] Step 8: Mixing the compound 9, a (2S,3R,4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain compound 10;

[0035] Step 9: Under a protective atmosphere, the compound 10 and an iodine organic solution are mixed to perform carbon-sulfur bond cyclization to obtain the compound 11;

[0036] Step 10: Mixing the compound 11 with an organic solvent to carry out a deprotection reaction to obtain the compound 12;

[0037] Step 11: Mixing the compound 12, diisopropylethylamine, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and an organic solvent to carry out a deprotection reaction and a condensation reaction to obtain compound 13;

[0038] Step 12: Mixing the compound 13, m-chloroperbenzoic acid and an organic solvent to carry out an oxidation reaction to obtain the compound 14;

[0039] Step 13:

[0040] When R is -OBn, the compound 14, ethanethiol and boron trifluoride ether solution are reacted to obtain a crude product, and the crude product is mixed with an alcoholic ammonia solution to perform a deprotection reaction to obtain the α-Amanitin;

[0041] When R is -H, the compound 14 is mixed with an alcoholic ammonia solution to carry out a deprotection reaction to obtain the Amaninamide.

[0042] Preferably, in step 1, the molar ratio of compound 2, acetyl chloride and pyridine is 1:1-3:1-5; the molar ratio of compound 2, N,N-dimethylpyridine and di-tert-butyl dicarbonate is 1:0.1-1:1-5.

[0043] Preferably, in step 2, the molar ratio of compound 3, N-Fmoc-asparagine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1;

[0044] In step 3, the molar ratio of compound 4, N-Fmoc-cysteine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1;

[0045] In step 4, the molar ratio of compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1;

[0046] In step 5, the molar ratio of compound 6, N-Fmoc-isoleucine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1;

[0047] In step 6, the molar ratio of compound 7, N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1;

[0048] In step 7, the molar ratio of compound 8, compound 15, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1;

[0049] In step 8, the molar ratio of compound 9, (2S,3R,4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1.

[0050] Preferably, in step 9, the molar ratio of compound 10 to elemental iodine in the organic iodine solution is 1:4, and the concentration of the organic iodine solution is 2 mg / mL.

[0051] Preferably, in step 10, the organic solvent is a diethylamine dichloromethane solution, the mass fraction of diethylamine in the diethylamine dichloromethane solution is 50%, and the concentration of compound 11 in the diethylamine dichloromethane solution is 0.1 g / mL.

[0052] Preferably, in step 11, the molar ratio of compound 12, diisopropylethylamine, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate is 1:2.2:2.

[0053] Preferably, the concentration of the alcoholic ammonia solution in step 13 is 7 mol / L.

[0054] Preferably, the acetylation reaction in step 1 is carried out at a temperature of 0° C. and for 2 hours.

[0055] Preferably, the esterification reaction in step 1 is carried out at room temperature and for 0.5 h.

[0056] Preferably, the condensation reaction and deprotection reaction in step 2 are carried out at room temperature and for 12 hours.

[0057] Preferably, the condensation reaction and deprotection reaction in step 3 are carried out at room temperature and for 12 hours.

[0058] Preferably, the condensation reaction and deprotection reaction in step 4 are carried out at room temperature and for 12 hours.

[0059] Preferably, the condensation reaction and deprotection reaction in step 5 are carried out at room temperature and for 12 hours.

[0060] Preferably, the condensation reaction and deprotection reaction in step 6 are carried out at room temperature and for 12 hours.

[0061] Preferably, the condensation reaction and deprotection reaction in step 7 are carried out at room temperature and for 12 hours.

[0062] Preferably, the condensation reaction and deprotection reaction in step 8 are carried out at room temperature and for 12 hours.

[0063] Preferably, the temperature for carbon-sulfur ring closure in step 9 is room temperature and the time is 1 hour.

[0064] Preferably, the deprotection reaction in step 10 is carried out at room temperature and for 1 hour.

[0065] Preferably, the condensation reaction and deprotection reaction in step 11 are carried out at room temperature and for 12 hours.

[0066] Preferably, the oxidation reaction in step 12 is carried out at room temperature and for 0.5 h.

[0067] Preferably, after the acetylation reaction in step 1 is completed, the reaction is quenched with a saturated ammonium chloride solution, the pH of the resulting reaction solution is adjusted to 2 with dilute hydrochloric acid, and extracted three times with dichloromethane. The organic phases are combined, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product.

[0068] Preferably, after the esterification reaction in step 1 is completed, the obtained product is decompressed to remove tert-butanol to obtain a crude product, the crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 hour, the solvent is removed under reduced pressure, and the product is separated by column chromatography to obtain the compound 3.

[0069] Preferably, after the condensation reaction and deprotection reaction in step 2 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The product is separated by column chromatography to obtain the compound 4.

[0070] Preferably, after the condensation reaction and deprotection reaction in step 3 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The product is separated by column chromatography to obtain the compound 5.

[0071] Preferably, after the condensation reaction and deprotection reaction in step 4 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The product is separated by column chromatography to obtain the compound 6.

[0072] Preferably, after the condensation reaction and deprotection reaction in step 5 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The product is separated by column chromatography to obtain the compound 7.

[0073] Preferably, after the condensation reaction and deprotection reaction in step 6 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 h, and the solvent is removed under reduced pressure. The product is separated by column chromatography to obtain the compound 8.

[0074] Preferably, after the condensation reaction and deprotection reaction in step 7 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The product is separated by column chromatography to obtain the compound 9.

[0075] Preferably, after the condensation reaction and deprotection reaction in step 8 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product, which is then separated by column chromatography to obtain compound 10.

[0076] Preferably, after the carbon-sulfur ring-closure reaction in step 9 is completed, N,N-dimethylformamide is removed under reduced pressure and recovered to obtain a crude product, and the crude product is separated by column chromatography to obtain compound 11.

[0077] Preferably, after the deprotection reaction in step 10 is completed, the solvent is removed under reduced pressure, and the obtained crude product is separated by column chromatography to obtain the compound 12.

[0078] This application also provides an intermediate obtained by the preparation method described in the above technical solution, the structure of which is as follows:

[0079] In the compounds 9 to 14, R is -H or -OBn.

[0080] The present application also provides the use of the intermediate described in the above technical solution in the preparation of cyclic peptide toxin compounds.

[0081] The present application provides a method for preparing cyclic peptide toxin compounds α-Amanitin and Amaninamide. Compared with the prior art, the present application has the following beneficial effects:

[0082] The present application provides a total synthesis method for cyclic peptide toxin compounds α-Amanitin and Amaninamide. The raw materials and reagents used are easily purchased through commercial channels, and the raw material cost is low. The intermediates obtained in the synthesis method are stable, the reaction conditions are mild, the operation process is simple, the separation and purification process has good operability and high yield, and has important reference and practical value. It can achieve gram-scale preparation of α-Amanitin and Amaninamide. The preparation method of the present application has significant application value in the field of cyclic peptide toxin synthesis.

[0083] The present application also provides a series of intermediates that help promote the synthesis of cyclic peptide toxins and other polypeptide compounds, and have significant application value in the field of polypeptide synthesis. DETAILED DESCRIPTION

[0084] The present application provides a method for preparing cyclic peptide toxin compounds α-Amanitin and / or Amaninamide, wherein the α-Amanitin has a structure shown in Formula 1a, and the Amaninamide has a structure shown in Formula 1b:

[0085] The following steps are involved:

[0086] Step 1: subjecting compound 2 to acetylation and esterification to obtain compound 3;

[0087] Step 2: subjecting the compound 3 to a condensation reaction and a deprotection reaction to obtain compound 4;

[0088] Step 3: subjecting the compound 4 to condensation reaction and deprotection reaction to obtain compound 5;

[0089] Step 4: subjecting the compound 5 to condensation reaction and deprotection reaction to obtain compound 6;

[0090] Step 5: subjecting the compound 6 to condensation reaction and deprotection reaction to obtain compound 7;

[0091] Step 6: subjecting the compound 7 to condensation reaction and deprotection reaction to obtain compound 8;

[0092] Step 7: subjecting the compound 8 to condensation reaction and deprotection reaction to obtain compound 9;

[0093] Step 8: subjecting the compound 9 to condensation reaction and deprotection reaction to obtain compound 10;

[0094] Step 9: Compound 10 is subjected to carbon-sulfur ring closure in an iodine-mediated manner to obtain compound 11;

[0095] Step 10: subjecting the compound 11 to a deprotection reaction to obtain compound 12;

[0096] Step 11: subjecting the compound 12 to a deprotection reaction and a condensation reaction to obtain compound 13;

[0097] Step 12: subjecting the compound 13 to an oxidation reaction to obtain compound 14;

[0098] Step 13: subjecting the compound 14 to a deprotection reaction to obtain the cyclic peptide toxin compounds α-Amanitin and Amaninamide;

[0099] The structures of compounds 2 to 14 are as described above.

[0100] In this application, unless otherwise specified, the raw materials used are commercially available products in the field or are prepared using preparation methods well known to those skilled in the art.

[0101] In this application, the principle of the preparation method is shown in the following formula:

[0102] In the present application, Fmoc in the compound is fluorenylmethyloxycarbonyl, Ac is acetyl, Trt is trityl, and Bn is benzyl.

[0103] In the present application, the preparation method preferably comprises the following steps:

[0104] Step 1: Compound 2, pyridine, acetyl chloride and an organic solvent are mixed for acetylation reaction, and the obtained acetylated product, N,N-dimethylpyridine and di-tert-butyl dicarbonate are mixed for esterification reaction to obtain Compound 3;

[0105] Step 2: mixing the compound 3, N-Fmoc-asparagine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 4;

[0106] Step 3: mixing the compound 4, N-Fmoc-cysteine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 5;

[0107] Step 4: mixing the compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 6;

[0108] Step 5: mixing the compound 6, N-Fmoc-isoleucine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain the compound 7;

[0109] Step 6: Mixing the compound 7, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 8;

[0110] Step 7: Mixing the compound 8, compound 15, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent for condensation reaction and deprotection reaction to obtain the compound 9; the compound 15 is N-Fmoc-6-benzyloxytryptophan or N-Fmoc-tryptophan;

[0111] Step 8: Mixing the compound 9, a (2S,3R,4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain compound 10;

[0112] Step 9: Under a protective atmosphere, the compound 10 and an iodine organic solution are mixed to perform carbon-sulfur bond cyclization to obtain the compound 11;

[0113] Step 10: Mixing the compound 11 with an organic solvent to carry out a deprotection reaction to obtain the compound 12;

[0114] Step 11: Mixing the compound 12, diisopropylethylamine, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and an organic solvent to carry out a deprotection reaction and a condensation reaction to obtain compound 13;

[0115] Step 12: Mixing the compound 13, m-chloroperbenzoic acid and an organic solvent to carry out an oxidation reaction to obtain the compound 14;

[0116] Step 13:

[0117] When R is -OBn, the compound 14, ethanethiol and boron trifluoride ether solution are reacted to obtain a crude product, and the crude product is mixed with an alcoholic ammonia solution to perform a deprotection reaction to obtain the α-Amanitin;

[0118] When R is -H, the compound 14 is mixed with an alcoholic ammonia solution to carry out a deprotection reaction to obtain the Amaninamide.

[0119] In this application, unless otherwise specified, the raw materials used are commercially available products in the field.

[0120] In the present application, the compound 2, pyridine, acetyl chloride and an organic solvent are mixed for acetylation reaction, and the obtained acetylated product, N,N-dimethylpyridine and di-tert-butyl dicarbonate are mixed for esterification reaction to obtain the compound 3.

[0121] In the present application, the molar ratio of compound 2, acetyl chloride and pyridine in step 1 is preferably 1:1-3:1-5, more preferably 1:1.5:1.5.

[0122] In the present application, the organic solvent is preferably dichloromethane, more preferably dry dichloromethane.

[0123] In the present application, the temperature of the acetylation reaction is preferably 0° C., and the time is preferably 2 h.

[0124] The present application preferably dissolves compound 2 in dry dichloromethane, and adds the pyridine and acetyl chloride under ice bath conditions.

[0125] After the acetylation reaction is completed, the present application preferably quenches the reaction with a saturated ammonium chloride solution, adjusts the pH of the resulting reaction solution to 2 with dilute hydrochloric acid, extracts three times with dichloromethane, combines the organic phases, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is recorded as the acetylation reaction.

[0126] In the present application, the molar ratio of the compound 2, N,N-dimethylpyridine and di-tert-butyl dicarbonate is preferably 1:0.1 to 1:1 to 5, and more preferably 1:0.3:2.

[0127] In the present application, the temperature of the esterification reaction is preferably room temperature, and the time is preferably 0.5 h.

[0128] The present application preferably dissolves the crude product in tert-butanol, adds N,N-dimethylpyridine and di-tert-butyl dicarbonate, and performs the esterification reaction.

[0129] After the esterification reaction is completed, the present application preferably removes tert-butanol from the obtained product under reduced pressure to obtain a crude product, and the crude product is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The product is separated by column chromatography to obtain the compound 3.

[0130] After obtaining the compound 3, the present application mixed the compound 3, N-Fmoc-asparagine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 4.

[0131] In the present application, the molar ratio of compound 3, N-Fmoc-asparagine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is preferably 1:1.1:1.1:1.1;

[0132] In the present application, the organic solvent is preferably dry dichloromethane.

[0133] In the present application, the temperature of the condensation reaction and the deprotection reaction is preferably room temperature, and the time is preferably 12 hours.

[0134] The present application preferably dissolves the compound 3 in dry dichloromethane, adds N-Fmoc-asparagine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with a saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The mixture is separated by column chromatography to obtain the compound 4.

[0135] After obtaining compound 4, the present application mixes the compound 4, N-Fmoc-cysteine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain compound 5.

[0136] In the present application, the molar ratio of compound 4, N-Fmoc-cysteine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is preferably 1:1.1:1.1:1.1;

[0137] In the present application, the organic solvent is preferably dry dichloromethane.

[0138] In the present application, the temperature of the condensation reaction and the deprotection reaction is preferably room temperature, and the time is preferably 12 hours.

[0139] The present application preferably dissolves the compound 4 in dry dichloromethane, adds N-Fmoc-cysteine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with a saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, the solvent is removed under reduced pressure, and separated by column chromatography to obtain the compound 5.

[0140] After obtaining compound 5, the present application mixed the compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain compound 6.

[0141] In the present application, the molar ratio of compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is preferably 1:1.1:1.1:1.1.

[0142] In the present application, the organic solvent is preferably dry dichloromethane.

[0143] In the present application, the temperature of the condensation reaction and the deprotection reaction is preferably room temperature, and the time is preferably 12 hours.

[0144] The present application preferably dissolves the compound 5 in dry dichloromethane, adds N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with a saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, the solvent is removed under reduced pressure, and separated by column chromatography to obtain the compound 6.

[0145] After obtaining compound 6, the present application mixed the compound 6, N-Fmoc-isoleucine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain compound 7.

[0146] In the present application, the molar ratio of compound 6, N-Fmoc-isoleucine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is preferably 1:1.1:1.1:1.1.

[0147] In the present application, the organic solvent is preferably dry dichloromethane.

[0148] In the present application, the temperature of the condensation reaction and the deprotection reaction is preferably room temperature, and the time is preferably 12 hours.

[0149] The present application preferably dissolves the compound 6 in dry dichloromethane, adds N-Fmoc-isoleucine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with a saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, the solvent is removed under reduced pressure, and separated by column chromatography to obtain compound 7.

[0150] After obtaining compound 7, the present application mixed the compound 7, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 8.

[0151] In the present application, the molar ratio of compound 7, N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is preferably 1:1.1:1.1:1.1.

[0152] In the present application, the organic solvent is preferably dry dichloromethane.

[0153] In the present application, the temperature of the condensation reaction and the deprotection reaction is preferably room temperature, and the time is preferably 12 hours.

[0154] The present application preferably dissolves the compound 7 in dry dichloromethane, adds N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with a saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, the solvent is removed under reduced pressure, and separated by column chromatography to obtain compound 8.

[0155] After obtaining compound 8, the present application mixes the compound 8, compound 15, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent for condensation reaction and deprotection reaction to obtain the compound 9; the compound 15 is N-Fmoc-6-benzyloxytryptophan or N-Fmoc-tryptophan.

[0156] In the present application, the molar ratio of compound 8, compound 15, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is preferably 1:1.1:1.1:1.1.

[0157] In the present application, the organic solvent is preferably dry dichloromethane.

[0158] In the present application, the temperature of the condensation reaction and the deprotection reaction is preferably room temperature, and the time is preferably 12 hours.

[0159] In this application, the structure of compound 15 is shown below:

[0160] Wherein R is -OBn or -H. When R is -OBn, the compound 15 is N-Fmoc-6-benzyloxytryptophan. When R is -H, the compound 15 is N-Fmoc-tryptophan.

[0161] The present application preferably dissolves the compound 8 in dry dichloromethane, adds N-Fmoc-6-benzyloxytryptophan, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with a saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The mixture is separated by column chromatography to obtain the compound 9a.

[0162] The present application preferably dissolves the compound 8 in dry dichloromethane, adds N-Fmoc-tryptophan, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with a saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is dissolved in a dichloromethane / diethylamine solution (the mass fraction of dichloromethane in the dichloromethane / diethylamine solution is preferably 50%), stirred at room temperature for 1 hour, and the solvent is removed under reduced pressure. The mixture is separated by column chromatography to obtain the compound 9b.

[0163] After obtaining compound 9, the present application mixes the compound 9, a (2S,3R,4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out a condensation reaction and a deprotection reaction to obtain compound 10.

[0164] In the present application, the molar ratio of compound 9, (2S,3R,4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is preferably 1:1.1:1.1:1.1.

[0165] In the present application, the organic solvent is preferably dry dichloromethane.

[0166] In the present application, the temperature of the condensation reaction and the deprotection reaction is preferably room temperature, and the time is preferably 12 hours.

[0167] In the present application, the structure of the (2S,3R,4R)-4,5-dihydroxyisoleucine derivative is shown in Formula 16 (Compound 16):

[0168] The present application preferably dissolves compound 9 in dry dichloromethane, adds (2S,3R,4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride under ice bath conditions, reacts at room temperature for 12 hours, washes the organic phase with saturated sodium carbonate solution, and dries over anhydrous sodium sulfate. The solvent is removed under reduced pressure to obtain a crude product, which is separated by column chromatography to obtain compound 10.

[0169] After obtaining compound 10, the present application mixed the compound 10 with an organic solution of iodine under a protective atmosphere to perform carbon-sulfur bond cyclization to obtain compound 11.

[0170] In the present application, the protective atmosphere is preferably nitrogen.

[0171] In the present application, the molar ratio of the compound 10 to the iodine element in the iodine organic solution is preferably 1:4, and the concentration of the iodine organic solution is preferably 2 mg / mL.

[0172] In the present application, the solvent of the organic solution of iodine is preferably N,N-dimethylformamide.

[0173] In the present application, the temperature for the carbon-sulfur ring closure is preferably room temperature, and the time is preferably 1 hour.

[0174] In the present application, the compound 10 is preferably added to an iodine-containing N,N-dimethylformamide solution under a nitrogen environment, reacted at room temperature for 1 hour, and the N,N-dimethylformamide is removed under reduced pressure and recovered to obtain a crude product, which is then separated by column chromatography to obtain compound 11.

[0175] After obtaining compound 11, the present application mixed the compound 11 with an organic solvent to carry out a deprotection reaction to obtain compound 12.

[0176] In the present application, the organic solvent is preferably a diethylamine dichloromethane solution, the mass fraction of diethylamine in the diethylamine dichloromethane solution is preferably 50%, and the concentration of the compound 11 in the diethylamine dichloromethane solution is preferably 0.1 g / mL.

[0177] In the present application, the temperature of the deprotection reaction is preferably room temperature, and the time is preferably 1 h.

[0178] The present application preferably dissolves the compound 11 in a dichloromethane / diethylamine solution, stirs at room temperature for 1 hour, removes the solvent under reduced pressure, and separates the obtained crude product by column chromatography to obtain the compound 12.

[0179] After obtaining compound 12, the present application mixed the compound 12, diisopropylethylamine, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and an organic solvent to carry out deprotection reaction and condensation reaction to obtain compound 13.

[0180] In the present application, the organic solvent is preferably N,N-dimethylformamide.

[0181] In the present application, the molar ratio of compound 12, diisopropylethylamine, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate is preferably 1:2.2:2.

[0182] In the present application, the temperature of the deprotection reaction and the condensation reaction is preferably room temperature, and the time is preferably 12 hours.

[0183] The present application preferably dissolves the compound 12 in trifluoroacetic acid to obtain a system with a concentration of compound 12 of 0.1 g / mL. The mixture is stirred at room temperature for 1 h, and the trifluoroacetic acid is removed under reduced pressure to obtain a crude product. The crude product is dissolved in dry N,N-dimethylformamide, and diisopropylethylamine and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate are added. The mixture is reacted at room temperature for 12 h, and the N,N-dimethylformamide is removed under reduced pressure and recovered to obtain a crude product, which is separated by column chromatography to obtain compound 13.

[0184] After obtaining compound 13, the present application mixes the compound 13, m-chloroperbenzoic acid and an organic solvent to carry out an oxidation reaction to obtain compound 14.

[0185] In the present application, the molar ratio of compound 13 to m-chloroperbenzoic acid is preferably 1:1.

[0186] In the present application, the organic solvent is preferably a mixed solution of isopropanol and ethanol, and the volume ratio of isopropanol to ethanol in the mixed solution of isopropanol and ethanol is preferably 2:1.

[0187] In the present application, the temperature of the oxidation reaction is preferably room temperature, and the time is preferably 0.5 h.

[0188] The present application preferably dissolves the compound 13 in a mixed solution of isopropanol and ethanol to obtain a system with a concentration of compound 13 of 0.2 g / mL, adds m-chloroperbenzoic acid, reacts at room temperature for 0.5 h, removes the solvent under reduced pressure to obtain a crude product, and separates the crude product by column chromatography to obtain compound 14.

[0189] After obtaining compound 14, when R is -OBn (ie, compound 14a), the present application reacts the compound 14, ethanethiol, and boron trifluoride ether solution to obtain a crude product, and the crude product is mixed with an alcoholic ammonia solution for deprotection reaction to obtain the α-Amanitin.

[0190] The present application preferably dissolves the compound 14a in ethyl mercaptan, adds a solution of boron trifluoride in ethyl ether, reacts at room temperature for 1 hour, removes ethyl mercaptan under reduced pressure to obtain a crude product, dissolves the crude product in a solution of ammonia in methanol, reacts at room temperature for 2 hours, removes the solvent under reduced pressure, and separates the obtained crude product by column chromatography to obtain compound 1a.

[0191] In the present application, the molar ratio of compound 14a to boron trifluoride in boron trifluoride ether solution is preferably 1:20; the concentration of compound 14a in ethanethiol is preferably 8 mg / mL; and the concentration of the crude product in ammonia methanol solution is preferably 0.01 g / mL.

[0192] When R is -H (ie, compound 14b), the present application mixes the compound 14 with an alcoholic solution of ammonia to perform a deprotection reaction to obtain the Amaninamide.

[0193] The present application preferably dissolves the compound 14b in an alcoholic ammonia solution, reacts at room temperature for 2 h, removes the solvent under reduced pressure to obtain a crude product, and separates the crude product by column chromatography to obtain compound 1b.

[0194] In the present application, the concentration of the compound 14b in the alcoholic ammonia solution is preferably 0.01 g / mL.

[0195] In the present application, the concentration of the alcoholic ammonia solution is preferably 7 mol / L; the alcoholic ammonia solution is preferably a methanolic ammonia solution.

[0196] This application also provides a series of intermediates, the structures of which are as follows:

[0197] In the compounds 9 to 14, R is -H or -OBn.

[0198] The present application also provides a method for preparing the intermediate described in the above technical solution. The method for preparing the intermediate is consistent with the method for preparing compounds 4 to 14 in the above solution, and will not be repeated here.

[0199] The present application also provides the use of the intermediate described in the above technical solution in the preparation of cyclic peptide toxin compounds.

[0200] The following will be combined with the embodiments of this application to clearly and completely describe the technical solutions in this application. Obviously, the embodiments described are only some of the embodiments of this application, not all of them. Based on the embodiments of this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0201] Example 1: Synthesis of Compound 3, the principle is as follows:

[0202] Place a magnetic stir bar in a 2500 mL round-bottom flask. Add compound 2 (N-Fmoc-4-hydroxyproline) (35.31 g, 100 mmol) to the 2500 mL round-bottom flask. Add 200 mL of dry dichloromethane and stir to dissolve. Stir in an ice-water bath for 5 minutes. Add pyridine (11.86 g, 150 mmol) via syringe and stir for 5 minutes. Slowly add acetyl chloride (11.70 g, 150 mmol) dropwise via syringe. Incubate in an ice-water bath for 30 minutes and allow to react at room temperature for 1 hour. After the reaction is complete, adjust the pH to 2 with 1N hydrochloric acid and extract three times with 500 mL of dichloromethane. Combine the organic phases, dry over anhydrous sodium sulfate, filter, and remove the dichloromethane under reduced pressure to obtain the crude product. The crude product is used directly in the next synthetic reaction without further purification. Dissolve the crude product in 400 mL of tert-butyl alcohol, add DMAP (1.84 g, 30 mol%), and stir at room temperature. A solution of (Boc)2O (43.62 g, 200 mmol) dissolved in 100 mL of tert-butanol was added dropwise to the reaction mixture. The reaction was allowed to react at room temperature and monitored by TLC until the starting material was consumed. Upon completion of the reaction, the tert-butanol solvent was removed under reduced pressure to obtain a crude product. The crude product was dissolved in 150 mL of dichloromethane and the Fmoc protecting group was removed by adding 150 mL of diethylamine. The reaction was stopped by TLC monitoring until the starting material was completely consumed. The mixed solution of dichloromethane and diethylamine was removed under reduced pressure to obtain the crude product. The crude product was purified by chromatography on a 200-300 mesh silica gel column (ethyl acetate / petroleum ether = 40%-50% by volume) to obtain 15.46 g of a yellow oily liquid, with a yield of 67.5%. 1 H NMR (600MHz, DMSO) δ5.07(d,J=2.7Hz,1H),3.65(t,J=7.6Hz,1H),3.15(dd,J=1 2.0,5.2Hz,1H),2.76(dd,J=12.0,1.5Hz,1H),2.00-1.98(m,5H),1.40(s,9H). 13 C NMR(151MHz,DMSO)δ173.1,170.1,80.2,75.0,59.1,52.2,36.4,27.6,20.9.HRMS(ESI+)calcd.for C 11 H 20 NO4 + (M+H) + :230.1387; found:230.1399.

[0203] Example 2: Synthesis of Compound 4, the principle is as follows:

[0204] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add N-Fmoc-asparagine (65.60 g, 0.11 mol) to the 2500 mL round-bottom flask. Add 500 mL of dry dichloromethane and stir to dissolve. Incubate in an ice-water bath for 5 minutes. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12.70 g, 0.11 mol) and 1-hydroxybenzotriazole (14.90 g, 0.11 mol) and incubate in an ice-water bath for 30 minutes. Then, add compound 3 (35.70 g, 0.1 mol) dropwise and let it react in an ice-water bath for 30 minutes. Allow to react overnight at room temperature and monitor the reaction by TLC until the starting material is consumed. Wash the organic phase three times with saturated sodium bicarbonate and extract it three times with 1000 mL of dichloromethane. Combine the organic phases, wash with salt, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure to obtain the crude product. The crude product is used directly in the next deprotection reaction without purification. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane and stirred at room temperature. The reaction was stopped after TLC monitoring of the complete consumption of the starting material. The solvent and diethylamine were removed under reduced pressure, and the product was purified by chromatography on a 200-300 mesh silica gel column (ethyl acetate / petroleum ether = 50%-60% by volume) to afford 52.00 g of compound 4 as a white solid in an 89% yield. 1 H NMR (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,1 H),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). 13 C NMR(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.HRMS(ESI+)calcd.for C 34 H 40 N3O6 + (M+H) + :586.2912; found:586.2896.

[0205] Example 3: Synthesis of Compound 5, the principle is as follows:

[0206] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add N-Fmoc-cysteine ​​(64.40 g, 0.11 mol) to the 2500 mL round-bottom flask and stir to dissolve in 500 mL of dry dichloromethane. Incubate in an ice-water bath for 5 minutes. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12.70 g, 0.11 mol) and 1-hydroxybenzotriazole (14.90 g, 0.11 mol) and incubate in an ice-water bath for 30 minutes. Then, add compound 4 (70.70 mg, 0.11 mol) dropwise and incubate in an ice-water bath for 30 minutes. Allow to react overnight at room temperature and monitor the reaction by TLC until the starting material is consumed. Wash the organic phase with saturated sodium bicarbonate and extract three times with 1000 mL of dichloromethane. Combine the organic phases, wash with salt, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure to obtain the crude product. The crude product is used directly in the next deprotection reaction without purification. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane and stirred at room temperature. The reaction was stopped after TLC monitoring of the complete consumption of the starting material. The solvent and diethylamine were removed under reduced pressure, and the product was purified by chromatography on a 200-300 mesh silica gel column (ethyl acetate / petroleum ether = 75%-80% by volume) to afford 76.30 g of compound 5 as a white solid in an 82% yield. 1 H NMR (600MHz, DMSO) δ8.73 (s, 1H), 8.25 (d, J = 7.8Hz, 1H), 7.34 -7.29(m,12H),7.23(dd,J=15.4,7.2Hz,10H),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= 7.7,5.0Hz,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). 13 C NMR (151MHz, DMSO) δ172.8,170.1,169.9,169.6,168.3,144.7,144.5,129.1,128.5,127.9,127.4,126.7, 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.HRMS(ESI+)calcd.for C34 H 40 N3O6 + (M+H) + :931.4099; found:931.4084.

[0207] Example 4: Synthesis of Compound 6, the principle is as follows:

[0208] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add N-Fmoc-glycine (32.70 g, 0.11 mol) to the 2500 mL round-bottom flask and stir to dissolve in 500 mL of dry dichloromethane. Incubate in an ice-water bath for 5 minutes. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12.70 g, 0.11 mol) and 1-hydroxybenzotriazole (14.90 g, 0.11 mol) and incubate in an ice-water bath for 30 minutes. Then, add compound 5 (108.70 g, 0.11 mol) dropwise and incubate in an ice-water bath for 30 minutes. Allow to react overnight at room temperature and monitor the reaction by TLC until the starting material is consumed. Wash the organic phase with saturated sodium bicarbonate and extract three times with 1000 mL of dichloromethane. Combine the organic phases, wash with salt, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure to obtain the crude product. The crude product is used directly in the next deprotection reaction without purification. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane and stirred at room temperature. The reaction was stopped after TLC monitoring of the complete consumption of the starting material. The solvent and diethylamine were removed under reduced pressure, and the product was purified by chromatography on a 200-300 mesh silica gel column (methanol / dichloromethane volume ratio = 0% to 0.5%) to afford 68.10 g of compound 6 as a white solid, in a 69% yield. 1 H NMR (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=15.1,4.9Hz, 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). 13C NMR (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.for C 58 H 62 N5O8S + (M+H) + :988.4314; found:988.4307.

[0209] Example 5: Synthesis of Compound 7, the principle is as follows:

[0210] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add N-Fmoc-isoleucine (35.30 g, 0.11 mol) to the 2500 mL round-bottom flask. Add 500 mL of dry dichloromethane and stir to dissolve. Incubate in an ice-water bath for 5 minutes. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12.70 g, 0.11 mol) and 1-hydroxybenzotriazole (14.90 g, 0.11 mol) and incubate in an ice-water bath for 30 minutes. Then, add compound 6 (98.70 g, 0.10 mol) dropwise and incubate in an ice-water bath for 30 minutes. Allow to react overnight at room temperature and monitor the reaction by TLC until the starting material is consumed. Wash the organic phase with saturated sodium bicarbonate and extract three times with 1000 mL of dichloromethane. Combine the organic phases, wash with salt, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure to obtain the crude product. The crude product is used directly in the next deprotection reaction without purification. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane and stirred at room temperature. The reaction was stopped after the starting material was consumed by monitoring with TLC. The solvent and diethylamine were removed under reduced pressure, and the product was purified by chromatography on a 200-300 mesh silica gel column (methanol / dichloromethane volume ratio = 0%-1%) to afford 80.30 g of compound 7 as a white solid in a yield of 73%. 1H NMR (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,4 H),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 C NMR (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.for C 64 H 73 N6O9S + (M+H) + :1101.5104;found:1101.5122.

[0211] Example 6: Synthesis of Compound 8, the principle is as follows:

[0212] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add N-Fmoc-glycine (32.60 g, 0.11 mol) to the 2500 mL round-bottom flask. Add 500 mL of dry dichloromethane and stir to dissolve. Incubate in an ice-water bath for 5 minutes. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12.70 g, 0.11 mol) and 1-hydroxybenzotriazole (14.90 g, 0.11 mol) and incubate in an ice-water bath for 30 minutes. Then, add compound 7 (110.00 g, 0.10 mol) dropwise and incubate in an ice-water bath for 30 minutes. Allow to react overnight at room temperature and monitor the reaction by TLC until the starting material is consumed. Wash the organic phase with saturated sodium bicarbonate and extract three times with 1000 mL of dichloromethane. Combine the organic phases, wash with salt, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure to obtain the crude product. The crude product is used directly in the next deprotection reaction without purification. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane and stirred at room temperature. The reaction was stopped after TLC monitoring of the complete consumption of the starting material. The solvent and diethylamine were removed under reduced pressure, and the product was purified by chromatography on a 200-300 mesh silica gel column (methanol / dichloromethane volume ratio = 0% to 1.5%) to afford 70.60 g of compound 8 as a white solid in a 61% yield. 1H NMR (600MHz, DMSO) δ8.68 (s, 1H), 8.46 (d, J = 7.4Hz, 1H), 8.32 (d, J = 5.3Hz, 1H), 8.07 (d,J=8.0Hz,1H),7.97(s,1H),7.35-7.28(m,12H),7.25-7.23(m,10H),7.18(d,J=7. 7Hz,8H),5.19(s,1H),4.72(dd,J=13.7,7.0Hz,1H),4.46(dd,J=13.8,7.0Hz,1H),4. 26(s,1H),4.21(t,J=7.8Hz,1H),3.79-3.73(m,4H),3.45-3.30(m,1H),3.11(d,J=6. 6Hz,2H),2.65(dd,J=15.1,5.1Hz,1H),2.56(dd,J=15.2,7.4Hz,1H),2.50(s,1H),2. 40(dd,J=11.0,5.1Hz,1H),2.37-2.28(m,1H),2.22(t,J=9.3Hz,1H),2.10-2.03(m,1 H),1.98(d,J=8.6Hz,1H),1.90(d,J=6.2Hz,3H),1.76(t,J=14.7Hz,1H),1.50-1.43( m,1H),1.41(s,9H),1.10-1.00(m,1H),0.84(d,J=6.7Hz,3H),0.81(t,J=7.3Hz,3H). 13 C NMR (151MHz, DMSO) δ172.7,171.4,170.1,169.9,169.4,169.7,168.4,168.3,144.7,144.2,129.1,128.6,128.5,128.0,127.4,126.7,126.3, 80.6,72.3,69.5,65.7,58.1,56.5,51.7,51.0,47.8,44.5,41.8,37.9,37.1,34.2,33.8,27.6,24.3,20.7,15.4,11.2.HRMS(ESI+)calcd.for C 66 H 76 N7O 10 S + (M+H) + :1158.5369; found:1158.5354.

[0213] Example 7: Synthesis of compounds 9a and 9b, the principle is as follows:

[0214] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add N-Fmoc-6-benzyloxytryptophan (68.90 g, 0.10 mol) to the 2500 mL round-bottom flask. Add 500 mL of dry dichloromethane and stir to dissolve. Incubate in an ice-water bath for 5 minutes. Add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (12.70 g, 0.11 mol) and 1-hydroxybenzotriazole (14.90 g, 0.11 mol) and incubate in an ice-water bath for 30 minutes. Subsequently, add compound 8 (143.80 g, 0.11 mol) dropwise and incubate in an ice-water bath for 30 minutes. Allow to react overnight at room temperature and monitor the reaction by TLC until the starting material is consumed. Wash the organic phase with saturated sodium bicarbonate and extract three times with 1000 mL of dichloromethane. Combine the organic phases, wash with salt, dry over anhydrous sodium sulfate, filter, and remove the solvent under reduced pressure to obtain the crude product. The crude product was used directly in the next step of deprotection. The crude product was dissolved in a 50 wt% diethylamine solution in dichloromethane and stirred at room temperature. The reaction was stopped after TLC monitoring of the complete consumption of the starting material. The solvent and diethylamine were removed under reduced pressure, and the product was purified by chromatography on a 200-300 mesh silica gel column (methanol / dichloromethane by volume = 0%-2%) to afford 100.00 g of compound 9a as a white solid in a 69% yield. 1H NMR(600MHz,DMSO)δ10.65(s,1H),8.64(s,1H),8.46(d,J=7.4Hz,1H),8.24(d,J=5.5Hz,1H),8.18(s,1H),8.02(d,J=7.9Hz,1H),7.92(d,J=8.5Hz,1H),7.46(d,J=7.4Hz,2H),7.42(d,J=8.6Hz,1H),7.38(t,J=7.5Hz,2H),7.32-7.28(m,12H),7.24-7.22(m,10H),7.18-7.15(m,8H),7.10(d,J=7.8Hz,1H),7.04(s,1H),6.91(s,1H),6.72(d,J=8.6Hz,1H),5.17(s,1H),5.10-5.08(m,2H),4.73-4.67(m,1H),4.48-4.42(m,1H),4.24(t,J=7.7Hz,1H),4.19(t,J=7.9Hz,1H),3.82-3.78(m,1H),3.78-3.74(m,2H),3.70(s,2H),3.46(dd,J=8.6,4.1Hz,1H),3.07(dd,J=14.1,3.7Hz,1H),2.72-2.58(m,2H),2.56-2.52(m,1H),2.37(dd,J=11.1,5.1Hz,1H),2.33-2.27(m,1H),2.25-2.15(m,1H),2.05(dd,J=13.6,6.6Hz,1H),1.88(s,3H),1.73(s,2H),1.45(s,1H),1.40(s,9H),1.12-1.02(m,1H),0.83(d,J=6.7Hz,3H),0.78(t,J=7.2Hz,3H). 13C NMR (151MHz, DMSO) δ174.9,171.2,170.1,169.9,169.3,169.1,168.7,168.3,168.2,154.4,144.6,144.2,137.7,1 36.8,129.0,128.5,128.3,127.9,127.5,127.4,126.6,126.3,122.5,118.9,110.6,109.1,96.0,80.6,72.2,69.5, 65.7,58.1,56.9,55.2,51.6,50.9,47.8,41.9,41.8,37.9,36.7,34.2,33.8,30.9,27.6,24.2,20.7,15.3,11.1.HRMS(ESI+)calcd.for C 84 H 92 N9O 12 S + (M+H) + :1450.6581;found:1450.6591.

[0215] Referring to the synthesis method of compound 9a, compound 8 (143.80 g, 0.11 mol) was reacted with N-Fmoc-tryptophan (42.60 g, 0.10 mol), and the Fmoc protecting group was removed to obtain white solid compound 9b, 104.80 g, 78%. 1H NMR(600MHz,DMSO)δ10.87(s,1H),8.67(s,1H),8.49(d,J=6.8Hz,1H),8.28(s,1H),8.22(s,1H),8.06(d,J=7.5Hz,1H),7.96(d,J=8.1Hz,1H),7.56(d,J=7.6Hz,1H),7.36-7.31(m,13H),7.24-7.23(m,8H),7.18(d,J=6.9Hz,10H),7.12(d,J=7.4Hz,1H),7.07(t,J=7.2Hz,1H),6.97(t,J=7.1Hz,1H),5.18(s,1H),4.73(d,J=6.4Hz,1H),4.48(d,J=6.4Hz,1H),4.26(t,J=7.3Hz,1H),4.21(t,J=7.7Hz,1H),3.82-3.73(m,5H),3.54-3.47(m,1H),3.38(s,2H),3.19-3.08(m,1H),2.74(dd,J=13.8,9.0Hz,1H),2.64(d,J=10.5Hz,1H),2.56(dd,J=14.5,7.0Hz,1H),2.39(d,J=5.8Hz,1H),2.37-2.27(m,1H),2.22(s,1H),2.11-2.02(m,1H),1.94-1.84(m,3H),1.76(s,2H),1.47(s,1H),1.41(s,9H),1.15-1.04(m,1H),0.85(d,J=6.2Hz,3H),0.80(t,J=7.0Hz,3H). 13 C NMR(151MHz,DMSO)δ174.9,171.3,170.1,169.9,169.4,169.2,168.8,168.3,168.2,144.7,144.2,136.3,129.1,128.6,128.5,128.0,127.4,126.7,126.3,123.8,120.9,118.4,118.3,111.3,110.7,80.7,72.3,69.5,65.7,58.1,56.9,55.3,51.7,50.9,47.8,41.9,41.8,37.9,36.8,34.3,33.8,30.9,27.6,24.2,20.7,15.4,11.2.HRMS(ESI+)calcd.for C 77 H 86 N9O 11 S +(M+H) + :1344.6162; found:1344.6189.

[0216] Example 8: Synthesis of compounds 10a and 10b, the principle is as follows:

[0217] Place a magnetic stir bar in a 2500 mL round-bottom flask. Add compound 16 (2S,3R,4R)-4,5-dihydroxyisoleucine derivative (46.90 g, 0.10 mol) to the 2500 mL round-bottom flask. Add 500 mL of dry dichloromethane and stir to dissolve. Incubate in an ice-water bath for 5 minutes. Add 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) (12.70 g, 0.11 mol) and 1-hydroxybenzotriazole (14.90 g, 0.11 mol) and incubate in an ice-water bath for 30 minutes. Then, add compound 9a (183.10 g, 0.11 mol) dropwise and incubate in an ice-water bath for 30 minutes. Allow to react overnight at room temperature and monitor the reaction by TLC until the starting material is consumed. The organic phase was washed with saturated sodium bicarbonate and extracted three times with 1000 mL of dichloromethane. The combined organic phases were washed with salt, dried over anhydrous sodium sulfate, filtered, and the solvent removed under reduced pressure to obtain a crude product. The product was purified by chromatography on a 200-300 mesh silica gel column (methanol / dichloromethane volume ratio = 0%-2%) to afford 152.00 g of compound 10a as a white solid in an 80% yield. 1H NMR(600MHz,DMSO)δ10.66(s,1H),8.68(s,1H),8.51(s,1H),8.33(s,1H),8.28(s,1H),8.16-8.05(m,1H),8.07-8.03(m,2H),7.90-7.89(m,3H),7.75(dd,J=13.3,7.3Hz,2H),7.58(d,J=9.0Hz,1H),7.55-7.49(m,2H),7.47(d,J=7.2Hz,2H),7.45-7.37(m,5H),7.35-7.30(m,13H),7.26-7.24(d,J=6.7Hz,10H),7.20(d,J=7.0Hz,8H),7.14(d,J=7.5Hz,1H),7.10(s,1H),6.94(s,1H),6.76(d,J=8.2Hz,1H),5.20(s,1H),5.11-5.08(d,J=16.4Hz,3H),4.84-4.74(m,2H),4.71(d,J=5.5Hz,1H),4.55-4.46(m,2H),4.38(dd,J=24.5,10.2Hz,2H),4.31(s,1H),4.24(dt,J=14.8,7.4Hz,2H),4.19-4.11(m,1H),4.04(d,J=7.6Hz,1H),3.81(s,4H),3.75(s,2H),3.41(s,1H),3.17(d,J=11.0Hz,1H),3.05(s,1H),2.67(d,J=10.7Hz,1H),2.60(s,1H),2.42(s,1H),2.35(s,1H),2.23(s,1H),2.13-2.04(m,1H),1.98(dd,J=16.9,6.9Hz,6H),1.91(d,J=8.7Hz,3H),1.79(s,1H),1.47(d,J=26.7Hz,1H),1.42(s,9H),1.14(d,J=26.1Hz,1H),0.87(s,3H),0.80(t,J=20.4Hz,3H),0.76(d,J=5.5Hz,3H). 13C NMR (151MHz, DMSO) δ171.6,171.3,170.5,170.2,170.1,169.9,169.7,169.4,169.2,168.5,168.4,168.3,156.3,154.5,144.7,144.3,1 44.0,143.7,140.7,140.6,137.7,136.7,129.1,128.6,128.5,128.3,128.0,127.7,127.6,127.4,127.2,126.7,126.3,125.6,125.3,1 22.5,122.1,120.1,119.0,109.8,109.1,95.9,80.7,72.3,71.6,69.6,69.5,66.2,65.7,62.9,58.1,56.9,54.9,54.7,53.5,51.7,50.9 ,47.9,46.7,42.1,41.8,39.5,37.9,36.8,35.3,34.3,33.9,27.6,27.5,24.2,20.8,20.7,20.6,15.4,11.2,10.3.HRMS(ESI+)calcd.for C 109 H 116 N 10 NaO 19 S + (M+Na) + :1923.8031; found:1923.8068.

[0218] Referring to the synthesis method of compound 10a, compound 9b (180.00 g, 0.11 mol) and compound 16 (2S, 3R, 4R)-4,5-dihydroxyisoleucine derivative (46.90 g, 0.10 mol) were reacted, and the Fmoc protecting group was removed to obtain white solid compound 10b, 147.20 g, in a yield of 82%. 1H NMR(600MHz,DMSO)δ10.86(s,1H),8.70(s,1H),8.53(s,1H),8.36(s,1H),8.30(s,1H),8.09(s,1H),7.90(d,J=6.5Hz,3H),7.77(dd,J=16.4,6.9Hz,2H),7.67(d,J=7.2Hz,1H),7.60(d,J=8.0Hz,1H),7.43(s,2H),7.34(s,14H),7.28-7.19(m,20H),7.08(d,J=6.8Hz,1H),7.01(s,1H),5.74(s,2H),5.22(s,1H),4.86-4.69(m,3H), 4.53(s,2H),4.42(d,J=11.0Hz,2H),4.33(s,1H),4.26(d,J=7.4Hz,2H),4.17(s,1H),4.05(d,J=7.6Hz,1H),3.81(d,J=33.7Hz,6H),3.44(s,1H),3.24(d,J=10.3Hz,1H),3.12(s,1H),2.66(d,J=39.3Hz,2H),2.57-2.50(m,1H),2.42(d,J=46.2Hz,2H),2.24(s,1H),2.08(s,1H),2.00(d,J=11.4Hz,6H),1.91(s,3H),1.82(s,1H),1.51(s,1H),1.43(s,9H),1.14(s,1H),0.82(m,9H). 13C NMR (151MHz, DMSO) δ171.7,171.3,170.6,170.2,170.1,169.9,169.7,169.4,169.2,168.6,168.4,168.3,156.3,144.7,144.3, 144.1,143.7,140.8,140.7,136.2,129.1,128.6,128.0,127.7,127.6,127.4,127.2,126.7,126.4,125.6,125.3,123.7,120.8 ,120.1,118.4,118.2,111.3,109.9,80.7,72.3,71.6,69.6,66.2,65.8,63.0,58.2,56.9,54.9,54.7,53.5,51.7,51.0,47.9,4 6.7,42.2,41.9,39.5,38.0,36.8,35.4,34.3,33.9,28.0,27.6,24.3,20.8,20.7,20.6,15.4,11.2,10.4.HRMS(ESI+)calcd.for C 102 H 110 N 10 NaO 18 S + (M+Na) + :1817.7612; found:1817.7625.

[0219] Example 9: Synthesis of compounds 11a and 11b. The principle is as follows:

[0220] A 2500 mL round-bottom flask was placed with a magnetic stir bar and the atmosphere was replaced three times using the Schlenk technique. Compound 10a (57.00 g, 0.03 mol) was added to the 2500 mL round-bottom flask under N₂ atmosphere. 500 mL of dry N,N-dimethylformamide was added and dissolved with stirring. Stirring was continued at room temperature. A 2 mg / mL solution of elemental iodine in DMF was prepared and 4 equiv of the iodine solution was slowly added dropwise to the reaction flask using a syringe. The reaction was allowed to react at room temperature for 2 h. After quenching, an excess of sodium thiosulfate solution was added. The 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 removed under reduced pressure to yield the crude product. The crude product was purified by chromatography on a 200-300 mesh silica gel column (methanol / dichloromethane = 0%-2% by volume) to afford 30.60 g of compound 11a as a white solid in a 62% yield. 1H NMR(600MHz,DMSO)δ11.06(d,J=49.8Hz,1H),8.63(s,1H),8.40(s,1H),8.28(d,J=45.1Hz,1H),8.18-7.98(m,3H),7.89(s,2H),7.74-7.74(m, 2H),7.65-7.60(m,1H),7.52-7.32(m,10H),7.26-7.24(m,6H),7.16(t,J=13.4Hz,9H),6.85(s,1H),6.77(d,J=9.1Hz,1H),5.20(s,1H),5.13-5.09(m,2H),4.77(s,1H),4.69(s,1H),4.48-4.26(m,3H),4.25-4.21(m,2H),4.17-4.09(m,1H),4.03(s,2H),3.88(s,2H),3.71(d,J=20.8Hz,4H),3.32(s,2H),2.94(d,J=8.9Hz,2H),2.61(s,2H),2.53-2.46(m,1H),2.37(s,1H),2.28(s,1H),2.06(s,1H),2.01-1.83(m,10H),1.54(s,1H),1.39(s,9H),1.20-1.06(m,1H),0.96-0.60(m,9H). 13C NMR (151MHz, DMSO) δ172.1,172.1,171.4,170.5,170.4,170.2,170.1,169.9,169.9,169.6,168.7,168.6,168.3,156.3,155.3,152.3,144.7 ,144.6,144.0,143.7,143.6,140.7,140.6,139.4,137.7,137.5,132. 2,128.9,128.6,128.4,128.3,127.8,127.6,127.5,127.4,127.3,127 .1,126.3,125.5,125.3,123.8,121.8,121.4,120.1,116.8,111.6,10 9.7,95.3,80.7,72.5,71.5,70.0,69.5,69.4,66.2,62.9,59.7,58.8, 58.2,54.9,54.7,51.8,48.1,46.7,42.9,42.4,39.5,37.7,35.2,33.9,27.6,24.5,20.7,20.5,15.5,11.0,10.2,10.0.HRMS(ESI+)calcd.for C 90 H 100 N 10 NaO 19 S + (M+Na) + :1679.6779; found:1679.6795.

[0221] Referring to the synthesis method of compound 11a, a white solid compound 11b was obtained, 34.40 g, with a yield of 74%. 1H NMR(600MHz,DMSO)δ11.18(d,J=8.8Hz,1H),8.63(s,1H),8.42(s,1H),8.25(d,J=40.3Hz,1H),8.10(d,J=34.1Hz,1H),7.96(d,J=14.3Hz,1H),7.89(s,2H),7.73(d,J=6.4Hz,2H),7.47(d,J=9.2Hz,1H),7.41(d,J=3.0Hz,2H),7.37-7.28(m,3H),7.26-7.24(m,7H),7.18-7.15(m,9H),7.12-7.09(m,1H),7.03-6.94(m,1H),5.18(s,1H),4.98-4.83(m,1H),4.75(s,1H),4.67(d,J=5.9Hz,1H),4.41-4.29(m,3H),4.26-4.16(m,3H),4.16-4.04(m,1H),4.05-4.01(m,3H),3.89-3.86(d,J=15.7Hz,2H),3.70(d,J=18.8Hz,4H),3.18-3.16(m,1H),3.04-2.90(m,2H),2.62-2.50(m,2H),2.31(d,J=54.1Hz,2H),2.07(s,1H),1.99-1.97(m,5H),1.94-1.92(m,4H),1.89-1.84(m,1H),1.53(s,1H),1.37(s,9H),1.15-1.12(m,1H),0.90-0.81(m,9H). 13 C NMR(151MHz,DMSO)δ172.0,171.4,170.4,170.3,170.2,170.1,169.9,169.8,169.7,169.7,168.6,168.3,162.3,144.7,143.9,140.7,136.8,128.6,127.6,127.4,127.1,126.3,125.3,121.9,120.0,118.6,116.2,110.9,80.7,72.5,71.9,71.5,69.5,66.2,65.9,62.9,62.9,59.7,58.8,58.1,54.8,51.8,48.1,46.6,42.8,42.4,37.9,37.7,35.76,35.2,33.9,30.7,27.6,24.5,20.7,20.6,15.4,14.1,11.0,10.0.HRMS(ESI+)calcd.for C83 H 94 N 10 NaO 18 S + (M+Na) + :1573.6360; found:1573.6369.

[0222] Example 10: Synthesis of compounds 12a and 12b, the principle is shown in the following formula:

[0223] A 2500 mL round-bottom flask was placed with a magnetic stirrer. Compound 11a (49.70 g, 0.03 mol) was added to the 2500 mL round-bottom flask, along with an excess of 50 wt% diethylamine in dichloromethane. The mixture was allowed to react at room temperature for 2 h. Upon completion, the solvent was removed under reduced pressure to yield the crude product. Chromatography on a 200-300 mesh silica gel column (methanol / dichloromethane = 2%-4% by volume) afforded 34.80 g of compound 12a as a white solid, yielding 81%.

[0224] 1H NMR(600MHz,DMSO)δ11.10-10.94(m,1H),11.00(s,1H),8.60(s,1H),8.41-8.35(m,1H),8.05-8.02(m,1H),7.94(d,J=37.2Hz,1H),7.88-7.80(m,1H),7.78-7.68(m,1H),7.57(d,J=22.5Hz,1H),7.51(d,J=12.6Hz,1H),7.47(d,J=7.2Hz,1H),7.40(d,J=7.1Hz,2H),7.33(dd,J=13.5,6.7Hz,2H),7.26-724(m,6H),7.18-7.13(m,9H),6.84(s,1H),6.78-6.71(m,1H),5.17(s,1H),5.14-5.01(m,2H),4.71-4.59(m,2H),4.44-4.25(m,1H),4.21(t,J=7.9Hz,1H),4.06(dd,J=15.8,9.1Hz,2H),3.96-3.82(m,3H),3.69(d,J=16.5Hz,4H),3.28(s,2H),3.18(s,1H),2.98-2.81(m,2H),2.58(s,2H),2.28(s,1H),2.10-2.03(m,1H),2.02-1.86(m,10H),1.83-1.74(m,2H),1.53(s,1H),1.39(s,9H),1.14(d,J=6.8Hz,2H),0.76(m,9H). 13 C NMR(151MHz,DMSO)δ172.4,171.3,170.9,170.4,170.3,170.0,169.9,169.8,169.6,169.5,168.6,168.3,155.2,144.7,137.5,128.6,128.4,128.3,127.8,127.6,127.4,127.3,126.3,123.7,117.0,111.5,109.7,95.3,80.7,72.4,70.0,69.5,66.3,58.6,58.1,54.8,51.7,48.1,42.3,39.5,35.2,33.9,27.6,24.3,22.4,20.7,15.5,11.1,10.4,9.5.HRMS(ESI+)calcd.for C 75 H 91 N 10 O 17 S +(M+H) + :1435.6279; found:1435.6289.

[0225] Referring to the synthesis method of compound 12a, a white solid compound 12b was obtained, 22.32 g, with a yield of 84%. 1 H NMR (600MHz, DMSO) δ11.23(d,J=31.1Hz,1H),8.73(s,1H),8.61(d,J=30.2Hz,1H),8.41(s ,1H),8.33-8.24(m,1H),8.05-7.97(m,2H),7.89-7.79(m,1H),7.76-7.72(m,1H),7.68(d, J=7.3Hz,1H),7.57(s,1H),7.30-7.24(m,6H),7.20-7.12(m,9H),7.04-6.98(m,1H),5.19(s,1H),4.89-4.79(m,1H),4.73(s,1H),4.6 9-4.51(m,2H),4.28(d,J=11.4Hz,1H),4.21(d,J=3.5Hz,1H),4.15(d,J=7.1Hz,1H),4.09-3.99(m,1H),3.97-3.81(m,3H),3.78-3.59( m,4H),3.53-3.40(m,1H),3.27(d,J=11.6Hz,1H),3.23-3.11(m,1H),3.00-2.84(m,1H),2.74(s,1H),2.59(s,1H),2.36-2.24(m,1H),2 .15-1.99(m,4H),1.98-1.90(m,8H),1.84(d,J=14.4Hz,1H),1.57(d,J=7.2Hz,2H),1.39(s,9H),1.20-1.08(m,1H),0.96-0.64(m,9H). 13C NMR (151MHz, DMSO) δ173.6,171.8,171.6,171.4,170.8,170.6,170.6,170.5,170.31,170.2,170.1, 170.0,169.1,168.9,168.8,168.7,145.2,145.2,137.6,129.1,127.9,126.8,125.7,122.6,119.5,1 18.9,117.4,111.4,81.2,73.0,72.5,69.9,68.6,66.9,63.5,60.2,58.6,55.4,54.4,52.3,48.6,44. 2,42.6,38.1,35.9,35.6,34.4,28.1,24.4,22.9,21.2,16.1,11.7,10.0,8.7.HRMS(ESI+)calcd.for C 68 H 85 N 10 O 16 S + (M+H) + :1329.5860; found:1329.5867.

[0226] Example 11: Synthesis of compounds 13a and 13b, the principle is shown in the following formula:

[0227] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add compound 12a (28.70 g, 0.02 mol) and 200 mL of trifluoroacetic acid to the 2500 mL round-bottom flask. Stir at room temperature for 0.5 h. Remove the trifluoroacetic acid under reduced pressure to yield a crude product. The crude product is used directly in the next step without further purification. Dissolve the crude product in dry N,N-dimethylformamide and stir at zero temperature for 5 min. Then, add diisopropylethylamine (6.90 g, 2 equiv) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (15.20 g, 2 equiv). Warm to room temperature and react for 12 h. Upon completion of the reaction, remove the N,N-dimethylformamide under reduced pressure to yield the crude product. The crude product was purified by 200-300 mesh silica gel column chromatography (methanol / dichloromethane volume ratio = 4%-6%) to obtain white solid compound 13a, 13.56 g, yield 61%. 1H NMR(600MHz,DMSO)δ11.08(d,J=42.1Hz,1H),8.95(s,1H),8.57(s,1H),8.33(d,J=12.6Hz,1H),8.05(d,J= 11.8Hz,2H),8.00-7.93(m,1H),7.92-7.83(m,2H),7.52(d,J=7.1Hz,1H),7.46(t,J=7.2Hz,1H),7.40(t,J=7.4Hz,2H),7.33-7.31(m,2H),7.16(dd,J=17.6,8.8Hz,2H),6.82-6.76(m,1H),5.35(s,1H),5.15-5.07(m,3H),4.92-4.81(m,2H),4.75(s,1H),4.57(d,J=9.8Hz,1H),4.36-4.26(m,2H),4.24-4.17(m,1H),4.13-4.01(m,43H),3.89(dd,J=17.0,6.7Hz,1H),3.72(s,1H),3.52-3.42(m,2H),3.21-3.16(m,2H),3.05-2.91(m,3H),2.65-2.59(m,1H),2.26(s,1H),2.11(t,J=11.0Hz,1H),2.02(s,3H),1.99(s,3H),1.95(s,3H),1.56(s,2H),1.13-1.06(m,2H),0.95(dd,J=20.7,9.5Hz,3H),0.83(t,J=6.9Hz,3H),0.78(d,J=6.0Hz,3H). 13 C NMR(151MHz,DMSO)δ171.9,171.9,171.8,171.1,171.1,170.6,170.2,169.9,169.8,169.8,169.7,169.6,168.1,167.5,162.2,158.7,155.3,151.9,137.2,128.4,127.8,127.4,124.9,123.6,122.5,121.7,116.4,109.9,72.3,71.0,70.2,69.5,62.9,61.8,59.1,54.9,54.3,53.0,52.8,52.5,50.5,39.5,35.7,34.4,30.8,28.9,25.2,20.9,20.7,20.6,14.8,11.2,10.6.HRMS(ESI+)calcd.for C 52 H 66 N10 O 16 NaS + (M+Na) + :1141.4271; found:1141.4280.

[0228] Referring to the synthesis method of compound 13a, a white solid compound 13b was obtained, 6.25 g, with a yield of 62%. 1 H NMR (600MHz, DMSO) δ11.25(s,1H),8.97(s,1H),8.56(s,1H),8.35(s,1H),8.06(d,J=14.9Hz,2H),7.98( d,J=9.9Hz,1H),7.91(d,J=7.0Hz,2H),7.58(d,J=7.7Hz,1H),7.32(s,1H),7.26(d,J=7.9Hz,1H),7.12( t,J=7.3Hz,1H),7.05-6.96(m,1H),5.36(s,1H),5.18(d,J=13.7Hz,1H),4.91(s,1H),4.86(d,J=7.7Hz, 1H),4.77(d,J=8.0Hz,1H),4.59(t,J=9.2Hz,1H),4.37-4.28(m,2H),4.19(dd,J=18.4,7.9Hz,1H),4.06 (dd,J=14.9,9.6Hz,3H),3.90(dd,J=17.1,6.9Hz,1H),3.73(s,1H),3.46(d,J=17.8Hz,2H),3.29-3.19( m,1H),3.17(s,1H),3.09-2.98(m,2H),2.90(dd,J=14.0,7.3Hz,2H),2.77(d,J=8.1Hz,1H),2.73(s,1H) ,2.62(s,1H),2.27(d,J=6.4Hz,1H),2.20-2.07(m,1H),2.03(s,3H),2.00(s,3H),1.95(s,3H),1.56(d, J=6.9Hz,2H),1.10(d,J=6.4Hz,2H),0.95(d,J=6.5Hz,3H),0.83(d,J=6.8Hz,3H),0.79(d,J=6.0Hz,3H). 13C NMR(151MHz,DMSO)δ172.0,171.9,171.1,170.7,170.2,169.9,169.8,169.7,169.6, 168.2,167.5,136.5,127.0,124.3,122.3,120.1,118.7,116.1,111.2,72.3,71.2,6 2.9,61.9,59.1,54.9,53.0,52.9,52.5,50.5,42.3,41.4,38.3,34.9,34.4,34.3,33 .6,30.6,29.0,25.2,21.0,20.7,20.6,14.8,11.2,11.0,10.6.HRMS(ESI+)calcd.for C 45 H 60 N 10 NaO 15 S + (M+Na) + :1035.3853; found:1035.3869.

[0229] Example 12: Synthesis of Compound 14, the principle is shown in the following formula:

[0230] Place a magnetic stirrer in a 2500mL round-bottom flask. Add compound 13a (5.60g, 0.005mol) to the 2500mL round-bottom flask, followed by 300mL of a mixed solution of isopropanol and ethanol (volume ratio of isopropanol to ethanol = 2:1), and stir in an ice-water bath for 10 minutes. Add a mixed solution of m-chloroperbenzoic acid in isopropanol and ethanol dropwise to the reaction mixture, and allow to react at zero temperature for 30 minutes. After the reaction is complete, remove the isopropanol and ethanol mixture under reduced pressure to obtain a crude product. The crude product is purified by 200-300 mesh silica gel column chromatography (volume ratio of methanol to dichloromethane = 5%-6%) to obtain 3.90g of compound 14a as a white solid in a 70% yield.

[0231] Referring to the synthesis method of compound 14a, a white solid compound 14b was obtained, 3.00 g, with a yield of 73%. 1H NMR(600MHz,DMSO)δ11.67(s,1H),8.88(d,J=5.8Hz,1H),8.55(s,1H),8.30(s,1H),8.26(d,J=10.5Hz,2H),7.89(d,J=8.8Hz,1H),7.85(d,J=10.4Hz,1H),7.81(d,J=9.0Hz,1H),7.65(d,J=8.1Hz,1H),7.44-7.38(m,2H),7.19(t,J=7.6Hz,1H),7.09(t,J=7.4Hz,1H),5.33(s,1H),5.21(dd,J=19.6,8.7Hz,1H),5.04-4.99(m,1H),4.91(s,1H),4.85(dd,J=10.4,3.3Hz,1H),4.76(d,J=8.4Hz,1H),4.32-4.24(m,3H),4.07-4.00(m,3H),3.89(dd,J=17.3,7.1Hz,1H),3.70(d,J=4.0Hz,1H),3.48(d,J=17.0Hz,2H),3.41(d,J=18.1Hz,1H),3.25(dd,J=15.1,6.8Hz,2H),3.10(t,J=13.3Hz,1H),2.93(dd,J=10.1,6.6Hz,2H),2.79-2.69(m,1H),2.57-2.51(m,1H),2.24(dd,J=13.6,6.7Hz,1H),2.12(d,J=12.8Hz,1H),2.02(s,3H),1.98(s,3H),1.95(s,3H),1.58-1.50(m,2H),1.12-1.03(m,2H),0.92(d,J=7.0Hz,3H),0.82(t,J=7.2Hz,3H),0.79(d,J=6.5Hz,3H). 13C NMR(151MHz,DMSO)δ172.0,171.7,171.2,170.5,170.3,170.2,170.1,169.8,169.7, 169.6,167.9,166.9,137.4,132.3,127.2,123.2,121.3,119.5,112.3,110.8,72.3,7 1.1,62.9,61.9,59.1,52.9,52.6,52.4,50.7,49.9,42.3,41.1,39.5,36.8,34.7,34 .6,34.5,32.9,29.2,25.2,20.9,20.7,20.5,14.8,11.1,10.8.HRMS(ESI+)calcd.for C 45 H 60 N 10 O 16 NaS + (M+Na) + :1051.3802;found:1051.3816.

[0232] Example 13: Synthesis of Compound 1a, the principle is shown in the following formula:

[0233] A 2500 mL round-bottom flask was placed with a magnetic stirrer. Compound 14a (4.00 g, 0.0045 mol) was added to the 2500 mL round-bottom flask, followed by the addition of ethanethiol solution and boron trifluoride etherate. After 2 hours of reaction, the solvent was removed under reduced pressure to yield the crude product, which had been debenzylated. The crude product was then added to a 7N ammonia-methanol solution and stirred at room temperature for 2 hours. The reaction was monitored by TLC until the starting material was exhausted. The ammonia-methanol solution was removed under reduced pressure to yield 1.883 g of the product, α-Amanitin1a, in a 50% yield.

[0234] 1H NMR (600MHz, DMSO) δ11.25(s,1H),9.21(s,1H),8.77(t,J=5.5Hz,1H),8.56(s,1H),8.37(s,1H),8 .24(d,J=10.4Hz,1H),8.21(s,1H),7.92(d,J=8.9Hz,1H),7.84(d,J=8.6Hz,1H),7.67(d,J=9.8Hz, 1H),7.42(d,J=8.7Hz,1H),7.37(s,1H),6.75(s,1H),6.61(d,J=8.6Hz,1H),5.18(s,1H),5.09(dd ,J=19.7,8.4Hz,1H),4.98-4.92(m,1H),4.84(dd,J=9.8,4.2Hz,1H),4.65(s,1H),4.41-4.36(m,3H ),4.25(ddd,J=27.6,14.9,8.0Hz,2H),3.90(dd,J=17.3,7.3Hz,1H),3.81(d,J=9.3Hz,1H),3.73( d,J=11.1Hz,1H),3.68(d,J=4.4Hz,1H),3.45-3.44(m,3H),3.41(s,1H),3.32-3.27(m,1H),3.23-3 .18(m,1H),3.17-3.15(m,1H),3.07(t,J=13.2Hz,1H),2.98-2.91(m,2H),2.74(t,J=13.8Hz,1H),2 .22-2.15(m,2H),1.86(t,J=11.0Hz,1H),1.59-1.50(m,2H),1.15-1.06(m,1H),0.85-0.77(m,9H). 13 C NMR (151MHz, DMSO) δ171.9,171.7,171.1,171.0,170.8,170.2,169.9,167.8,167.0,154.6,138.8,129.7,122.0,120.7,111.5,110.7,96. 7,71.9,68.6,63.5,62.1,59.2,55.8,54.9,53.9,52.7,50.8,50.0,4 8.6,42.4,41.3,37.8,37.3,34.6,33.4,29.2,25.2,14.7,11.4,10.7.

[0235] The synthesis principle of compound 1b is shown in the following formula:

[0236] Place a magnetic stirrer in a 2500 mL round-bottom flask. Add compound 14b (3.00 g, 0.003 mol) and 7N methanolic ammonia solution to the 2500 mL round-bottom flask. Stir at room temperature for 2 hours. Monitor the reaction by TLC until the starting material is consumed. Remove the methanolic ammonia solution under reduced pressure to obtain the crude product. Purify the crude product by 200-300 mesh silica gel column chromatography (methanol / dichloromethane = 10%-15% by volume) to obtain Amaninamide 1b as a white solid (1.92 g, 74% yield). 1 H NMR (600MHz, DMSO) δ11.80-11.03(m,1H),8.82(s,1H),8.57(s,1H),8.40(s,1H),8.29(d,J=10.3 Hz,1H),8.22(d,J=23.3Hz,1H),7.94(t,J=12.5Hz,1H),7.86(dd,J=26.5,7.9Hz,1H),7.69-7.62 (m,2H),7.40(d,J=7.7Hz,2H),7.18(dd,J=19.6,11.9Hz,1H),7.08(t,J=7.5Hz,1H),5.33-5.22( m,1H),5.16(dd,J=19.6,8.3Hz,1H),5.04-4.96(m,1H),4.85(dd,J=9.7,4.2Hz,1H),4.66(s,1H) ,4.44-4.369(m,3H),4.30-4.18(m,2H),3.90(dd,J=17.3,7.3Hz,1H),3.79(dd,J=29.6,10.1Hz, 2H),3.68(d,J=4.3Hz,1H),3.50-3.35(m,8H),3.31(dd,J=10.9,5.7Hz,8H),3.28-3.19(m,11H), 3.13(d,J=13.2Hz,1H),2.95(t,J=10.5Hz,2H),2.79(dd,J=27.3,10.7Hz,2H),2.23-2.16(m,2H) ,1.86(t,J=10.9Hz,1H),1.55(d,J=6.1Hz,2H),1.13-1.04(m,2H),0.80(dd,J=22.1,5.0Hz,9H). 13C NMR(151MHz,DMSO)δ171.9,171.7,171.21,171.0,170.8,170.2,169.9,167 .9,167.0,137.4,132.4,127.2,123.1,121.5,119.4,112.3,110.9,71.9,68 .7,63.5,62.10,59.2,58.9,55.8,54.0,52.6,50.9,49.9,42.4,41.7,41.0, 39.5,37.3,34.6,33.4,29.2,25.2,14.8,11.4,10.7.HRMS(ESI+)calcd.for C 39 H 54 N 10 NaO 13 S + (M+Na) + :925.3482; found:925.3489.

[0237] The above description is merely a preferred embodiment of the present application and does not constitute any form of limitation to the present application. It should be noted that those skilled in the art may make various improvements and modifications without departing from the principles of the present application, and such improvements and modifications shall also be considered within the scope of protection of the present application.

Claims

1. A method for preparing cyclic peptide toxin compounds α-Amanitin and / or Amaninamide, wherein the α-Amanitin has a structure shown in Formula 1a, and the Amaninamide has a structure shown in Formula 1b: It is characterized in that The following steps are involved: Step 1: subjecting compound 2 to acetylation and esterification to obtain compound 3; Step 2: subjecting the compound 3 to condensation reaction and deprotection reaction to obtain compound 4; Step 3: subjecting the compound 4 to condensation reaction and deprotection reaction to obtain compound 5; Step 4: subjecting the compound 5 to condensation reaction and deprotection reaction to obtain compound 6; Step 5: subjecting the compound 6 to condensation reaction and deprotection reaction to obtain compound 7; Step 6: subjecting the compound 7 to condensation reaction and deprotection reaction to obtain compound 8; Step 7: subjecting the compound 8 to condensation reaction and deprotection reaction to obtain compound 9; Step 8: subjecting the compound 9 to condensation reaction and deprotection reaction to obtain compound 10; Step 9: The compound 10 is subjected to carbon-sulfur bond cyclization in an iodine-mediated manner to obtain compound 11; Step 10: subjecting the compound 11 to a deprotection reaction to obtain a compound 12; Step 11: subjecting the compound 12 to a deprotection reaction and a condensation reaction to obtain a compound 13; Step 12: subjecting the compound 13 to an oxidation reaction to obtain a compound 14; Step 13: subjecting the compound 14 to a deprotection reaction to obtain the cyclic peptide toxin compounds α-Amanitin and Amaninamide; The structures of compounds 2 to 14 are as follows: In the compounds 9 to 14, R is -H or -OBn.

2. The preparation method according to claim 1, characterized in that: The following steps are involved: Step 1: mixing the compound 2, pyridine, acetyl chloride and an organic solvent for acetylation reaction, and mixing the obtained acetylated product with N,N-dimethylpyridine and di-tert-butyl dicarbonate for esterification reaction to obtain the compound 3; Step 2: mixing the compound 3, N-Fmoc-asparagine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent for condensation reaction and deprotection reaction to obtain the compound 4; Step 3: mixing the compound 4, N-Fmoc-cysteine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 5; Step 4: mixing the compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 6; Step 5: mixing the compound 6, N-Fmoc-isoleucine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 7; Step 6: mixing the compound 7, N-Fmoc-glycine, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain the compound 8; Step 7: Mixing the compound 8, the compound 15, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent for condensation reaction and deprotection reaction to obtain the compound 9; the compound 15 is N-Fmoc-6-benzyloxytryptophan or N-Fmoc-tryptophan; Step 8: Mixing the compound 9, (2S, 3R, 4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and an organic solvent to carry out condensation reaction and deprotection reaction to obtain compound 10; Step 9: Under a protective atmosphere, the compound 10 and an organic solution of iodine are mixed to perform carbon-sulfur bond cyclization to obtain the compound 11; Step 10: Mix the compound 11 and an organic solvent to carry out a deprotection reaction to obtain the compound 12; Step 11: mixing the compound 12, diisopropylethylamine, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and an organic solvent to carry out a deprotection reaction and a condensation reaction to obtain a compound 13; Step 12: Mix the compound 13, m-chloroperbenzoic acid and an organic solvent to carry out an oxidation reaction to obtain the compound 14; Step 13: When R is -OBn, the compound 14, ethanethiol and boron trifluoride ether solution are reacted to obtain a crude product, and the crude product is mixed with an alcohol solution of ammonia to perform a deprotection reaction to obtain the α-Amanitin; When R is -H, the compound 14 and an alcoholic solution of ammonia are mixed to carry out a deprotection reaction to obtain the Amaninamide.

3. The preparation method according to claim 2, characterized in that: In the step 1, the molar ratio of compound 2, acetyl chloride and pyridine is 1:1-3:1-5; the molar ratio of compound 2, N,N-dimethylpyridine and di-tert-butyl dicarbonate is 1:0.1-1:1-5.

4. The preparation method according to claim 2, characterized in that: In the step 2, the molar ratio of compound 3, N-Fmoc-asparagine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1; In the step 3, the molar ratio of compound 4, N-Fmoc-cysteine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1; In step 4, the molar ratio of compound 5, N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1; In the step 5, the molar ratio of compound 6, N-Fmoc-isoleucine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1; In step 6, the molar ratio of compound 7, N-Fmoc-glycine, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1; In step 7, the molar ratio of compound 8, compound 15, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.1; In step 8, the molar ratio of compound 9, (2S, 3R, 4R)-4,5-dihydroxyisoleucine derivative, 1-hydroxybenzotriazole and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride is 1:1.1:1.1:1.

1.

5. The preparation method according to claim 2, characterized in that: In the step 9, the molar ratio of compound 10 to the iodine element in the iodine organic solution is 1:4, and the concentration of the iodine organic solution is 2 mg / mL.

6. The preparation method according to claim 2, characterized in that: In the step 10, the organic solvent is a diethylamine dichloromethane solution, the mass fraction of diethylamine in the diethylamine dichloromethane solution is 50%, and the concentration of the compound 11 in the diethylamine dichloromethane solution is 0.1 g / mL.

7. The preparation method according to claim 2, characterized in that: In the step 11, the molar ratio of compound 12, diisopropylethylamine, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate is 1:2.2:

2.

8. The preparation method according to claim 2, characterized in that: The concentration of the alcohol solution of ammonia in step 13 is 7 mol / L.

9. The preparation method according to claim 1, characterized in that: The temperature of the acetylation reaction in step 1 is 0° C. and the time is 2 h.

10. The preparation method according to claim 1, characterized in that: The temperature of the esterification reaction in step 1 is room temperature and the time is 0.5 h.

11. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 2 is room temperature and the time is 12 hours.

12. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 3 is room temperature and the time is 12 hours.

13. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 4 is room temperature and the time is 12 hours.

14. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 5 is room temperature, and the time is 12 hours.

15. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 6 is room temperature, and the time is 12 hours.

16. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 7 is room temperature, and the time is 12 hours.

17. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 8 is room temperature and the time is 12 hours.

18. The preparation method according to claim 1, characterized in that: In step 9, the temperature for closing the carbon-sulfur bond is room temperature and the time is 1 hour.

19. The preparation method according to claim 1, characterized in that: The temperature of the deprotection reaction in step 10 is room temperature and the time is 1 hour.

20. The preparation method according to claim 1, characterized in that: The temperature of the condensation reaction and the deprotection reaction in step 11 is room temperature, and the time is 12 hours.

21. The preparation method according to claim 1, characterized in that: The temperature of the oxidation reaction in step 12 is room temperature and the time is 0.5 h.

22. The preparation method according to claim 1, characterized in that: After the acetylation reaction in step 1 is completed, the reaction is quenched with a saturated ammonium chloride solution, the pH of the resulting reaction solution is adjusted to 2 with dilute hydrochloric acid, and extracted with dichloromethane three times. The organic phases are combined, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product.

23. The preparation method according to claim 1, characterized in that: After the esterification reaction in step 1 is completed, the obtained product is decompressed to remove tert-butanol to obtain a crude product, the crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 hour, the solvent is removed under reduced pressure, and separated by column chromatography to obtain the compound 3.

24. The preparation method according to claim 1, characterized in that: After the condensation reaction and deprotection reaction in step 2 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 h, the solvent is removed under reduced pressure, and separated by column chromatography to obtain the compound 4.

25. The preparation method according to claim 1, characterized in that: After the condensation reaction and deprotection reaction in step 3 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 h, and the solvent is removed under reduced pressure. The compound 5 is separated by column chromatography to obtain the compound 5.

26. The preparation method according to claim 1, characterized in that: After the condensation reaction and deprotection reaction in step 4 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 h, and the solvent is removed under reduced pressure. The compound 6 is separated by column chromatography to obtain the compound 6.

27. The preparation method according to claim 1, characterized in that: After the condensation reaction and deprotection reaction in step 5 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 h, and the solvent is removed under reduced pressure. The compound 7 is separated by column chromatography to obtain the compound 7.

28. The preparation method according to claim 1, characterized in that: After the condensation reaction and deprotection reaction in step 6 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 h, and the solvent is removed under reduced pressure. The compound 8 is separated by column chromatography to obtain the compound 8.

29. The preparation method according to claim 1, characterized in that: After the condensation reaction and deprotection reaction in step 7 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is dissolved in a dichloromethane / diethylamine solution, stirred at room temperature for 1 h, and the solvent is removed under reduced pressure. The compound 9 is separated by column chromatography to obtain the compound 9.

30. The preparation method according to claim 1, characterized in that: After the condensation reaction and the deprotection reaction in step 8 are completed, the organic phase is washed with a saturated sodium carbonate solution, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure to obtain a crude product. The crude product is separated by column chromatography to obtain the compound 10.

31. The preparation method according to claim 1, characterized in that: After the carbon-sulfur ring-closure reaction in step 9 is completed, N,N-dimethylformamide is removed under reduced pressure and recovered to obtain a crude product, and the crude product is separated by column chromatography to obtain the compound 11.

32. The preparation method according to claim 1, characterized in that: After the deprotection reaction in step 10 is completed, the solvent is removed under reduced pressure, and the obtained crude product is separated by column chromatography to obtain the compound 12.

33. The intermediate obtained by the preparation method according to claim 1, characterized in that: The structure is as follows: In the compounds 9 to 14, R is -H or -OBn.

34. Use of the intermediate according to claim 33 in the preparation of cyclopeptide toxin compounds.

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