CN115677827A

CN115677827A - Peptide compounds

Info

Publication number: CN115677827A
Application number: CN202211432502.2A
Authority: CN
Inventors: 张利香; 吴丽芬; 马洪季; 李新宇; 付玉清; 余仕文; 岳泽乐
Original assignee: SHENZHEN JYMED TECHNOLOGY CO LTD
Current assignee: SHENZHEN JYMED TECHNOLOGY CO LTD
Priority date: 2022-11-16
Filing date: 2022-11-16
Publication date: 2023-02-03

Abstract

The present invention relates to compounds of formula I and the use of such compounds in solid phase synthesis, the compounds of formula I being useful in solid phase synthesis comprising a Lys-moiety linked to a peptide chain (AEEA) _n -gamma Glu-an intermediate of a polypeptide of a fatty alkanedioic acid side chain building block.

Description

Peptide compounds

Technical Field

The invention relates to a peptide compound and application thereof.

Background

GLP-1 receptor agonist and GLP-1/GIP double-receptor agonist are hot spots in research and development of medicaments for treating type II diabetes and are pursued by a plurality of international medicinal enterprises, such as Semaglutide (Semaglutide) and Tirzepatide. The number of amino acids of the peptide is about 40, and the Lys side chain is modified by PEG, glu and alkyl dicarboxylic acid, so that the hydrophilicity is greatly improved, and the binding force with albumin is enhanced; meanwhile, after mutation of 2 nd Ala at the N end into Aib, inactivation caused by DPP-IV enzymolysis is effectively avoided, the half-life period is remarkably prolonged, and the patient only needs to inject once every week, so that the medication compliance of the patient is greatly improved. In addition, the effects of the medicines in the aspects of losing weight and protecting the cardiovascular system are developed successively.

In the synthesis of such peptides comprising a Lys-moiety linked to the peptide chain (AEEA) _n In the industrial production of long peptide drugs of the-gamma Glu-fatty alkanedioic acid side chain construction unit, the traditional method mostly adopts an orthogonal protection strategy, and after the synthesis of the main chain is completed, the protecting group of the side chain connecting point is removed, and then the side chain amino acid coupling is carried out. Because the main chain sequence is longer and the side chain is larger, the traditional method can cause the main chain to be folded and curled, the amino-terminal amino acid and the side chain are difficult to be successfully connected, more side reactions are generated, more mismatched peptide impurities similar to the target product in structure are generated, the purification and the separation are very difficult, the good separation effect can not be achieved by using various purification and separation systems, if the separation is carried out for multiple times, the obvious loss of the product can be caused, the batch production yield is low, the production cost is high, and the industrialization can not be realized. Meanwhile, the existence of mismatched peptide impurities has adverse effects on the quality of the medicine, and not only influences the stability and effectiveness of the medicine, but also influences the safety of the medicine, thereby being harmful to the health of human bodies and not conforming to the regulatory requirements of national laws and regulations on the quality of the medicine.

Disclosure of Invention

To solve the problem of the prior art comprising a Lys-moiety linked to the peptide chain (AEEA) _n Problems in the Synthesis of peptides of the side chain building Block of-Gamma Glu-fatty alkanedioic acid, the present invention provides a novel intermediate peptide compound which can be easily inserted into a solid phase of a long peptide chainAnd (4) during synthesis.

The invention solves the technical problem and relates to the following compounds:

has a structure shown in a general formula I, and the structure is as follows:

wherein the content of the first and second substances,

r1 is selected from hydrogen, amino protecting group, COCH (CH 3) NH ₂ Amino protected COCH (CH 3) NH ₂ ，

R2 is selected from hydroxyl, hydroxyl protecting group, NHCH (CH) ₃ ) NHCH (CH) with COOH and carboxyl protected ₃ )COOH，

R3 is selected from hydrogen and ester protecting group,

r4 is selected from hydrogen, amino protecting group, CO (CH) ₂ ) _m R ₅ ，

m is 10 to 20, R ₅ Is COOH or carboxyl with a protecting group,

n is 1 or 2, and n is,

or enantiomers, diastereomers and salts thereof.

In some embodiments, R1 is selected from H, fmoc, dde, alloc, boc, moz, trt, dmb, mmt, mtt.

In some embodiments, R1 is selected from COCH (CH 3) NH ₂ 、COCH(CH3)NH ₂ -Fmoc、COCH(CH3)NH ₂ -Dde、COCH(CH3)NH ₂ -Alloc、COCH(CH3)NH ₂ -Boc、COCH(CH3)NH ₂ -Moz、COCH(CH3)NH ₂ -Trt、COCH(CH3)NH ₂ -Dmb、COCH(CH3)NH ₂ -Mmt、COCH(CH3)NH ₂ -Mtt。

In some embodiments, R2 is selected from OH, otBu, OCH ₃ 、OCH2CH ₃ 、OBzl、OCH ₂ CH＝CH ₂ 、OCH ₂ C ₆ H ₅ 。

In some embodiments, R2 is selected from NHCH (CH) ₃ )COOH、NHCH(CH ₃ )COOtBu、NHCH(CH ₃ )COOCH3、NHCH(CH ₃ )COOCH ₂ CH ₃ 、NHCH(CH ₃ )COOBzl、NHCH(CH ₃ )COOCH ₂ CH＝CH ₂ 、NHCH(CH ₃ )COOCH ₂ C ₆ H ₅ 。

In some embodiments, R3 is selected from H, tBu, CH ₃ 、CH ₂ CH ₃ 、Bzl、CH ₂ CH＝CH ₂ 、CH ₂ C ₆ H ₅ 。

In some embodiments, R4 is selected from H, fmoc, dde, alloc, boc, moz, trt, dmb, mmt or Mtt.

In some embodiments, m is 14 to 20.

Preferably, m is 16.

Preferably, m is 18.

In some embodiments, R5 is selected from COOH, COOtBu, COOCH ₃ 、COOCH ₂ CH ₃ 、COOBzl、COOCH ₂ CH＝CH ₂ 、COOCH ₂ C ₆ H ₅ 。

In some embodiments, R4 is selected from CO (CH) ₂ ) ₁₆ COOH、CO(CH ₂ ) ₁₈ COOH、CO(CH ₂ ) ₁₆ COOtBu、CO(CH ₂ ) ₁₈ COOtBu

In some embodiments, n is 2.

In some embodiments, R1 is Fmoc and R2 is NHCH (CH) ₃ ) COOH, R3 is tBu, R4 is CO (CH) ₂ ) ₁₈ COOtBu, n is 2.

In some embodiments, R1 is Alloc and R2 is NHCH (CH) ₃ ) COOH, R3 is Me, R4 is CO (CH) ₂ ) ₁₈ COOtBu, n is 2.

In some embodiments, R1 is NHCH (CH) ₃ ) CO-Fmoc, R2 is OH, R3 is tBu, R4 is CO (CH) ₂ ) ₁₆ COOtBu, n is 2.

In some embodiments, R1 is NHCH (CH) ₃ ) CO-Dde, R2 is OH, R3 is tBu, R4 is CO (CH) ₂ ) ₁₆ COOBzl, n is 2.

Peptide compounds having the structure of formula I are synthesized comprising a Lys-moiety linked to the peptide chain (AEEA) _n -gammaglu-fatty alkanedioic acid side chain building block.

In some embodiments, wherein the synthesis is solid phase peptide synthesis.

The application of the peptide compound with the structure shown in the general formula I in the preparation of antidiabetic polypeptide medicaments.

In some embodiments, the anti-diabetic polypeptide drug is a GLP-1 analog or a GIP/GLP-1 analog.

In some embodiments, the GLP-1 analog is Semaglutide.

In some embodiments, the GIP/GLP-1 analog is Tirzepatide.

Compared with the prior art, the invention has the following beneficial effects:

in the synthesis of peptides comprising a Lys-moiety linked to the peptide chain (AEEA) _n When the compound of the formula I is adopted, the side chain can be inserted more easily in solid phase peptide synthesis, so that the reaction rate of the compound I and the subsequent amino acid at the amino terminal of peptide resin is increased, the coupling efficiency is effectively increased, the generation of mismatching peptide impurities is reduced, the yield and the purity of a crude peptide product are greatly increased, the purification difficulty is reduced, the crude peptide is favorably purified and separated to obtain a refined peptide, the yield of the refined peptide can reach more than 30 percent, the purity is more than 99 percent, and the maximum single impurity can reach less than 0.2 percent. Through the technical scheme of this application, shorten production time, reduction in production cost improves production efficiency, is favorable to realizing the industrialization, and the reduction of mismatch peptide impurity simultaneously all has better effect to improving medicine quality, stability, security even.

Detailed Description

The present invention is further illustrated by the following examples, which are intended to be illustrative, but not limiting, of the present invention. It should be noted that various changes and modifications can be made by those skilled in the art without departing from the principle of the invention, and these changes and modifications also fall into the scope of the invention.

Example (b):

example 1

Fmoc-Ala-Lys(COOtBu(CH ₂ ) ₁₆ Preparation of CO-Glu (AEEA-AEEA) -OtBu) -OH

Structural formula (xvi):

(1)COOtBu(CH ₂ ) ₁₆ synthesis of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

weighing 40.0g of CTC resin (40 mmol, substitution value of 1.10 mmol/g), and adding into a polypeptide synthesis reaction column; weighing 30.8g of Fmoc-AEEA-OH, adding 200ml of DCM for dissolving until the mixture is clear, then adding 25.8g of DIEA, and fully stirring; then adding the mixed solution into a reaction column, stirring and reacting for more than 2.0h, and discharging waste.

6.4g of methanol and 12.9g of DIEA are weighed, 320ml of DCM is added and stirred evenly, and then a DCM solution of methanol/DIEA is added into a reaction column and stirred for reaction for more than 1.0h. Waste, DCM wash 4 times.

The reaction column was charged with 20% PIPE/DMF and the Fmoc protection was removed twice, the deprotection times were respectively 5min +15min and DMF washes 6 times. Weighing Fmoc-AEEA-OH (23.1g, 1.5eq.) and dissolving in 100ml DCM, adding DIC (11.4g, 2.25eq.) and activating at 0-10 ℃ for 4min, adding into a synthesis reaction column, and reacting for 2-4h at 25 +/-5 ℃ under the protection of nitrogen. Waste, DCM wash 4 times.

Repeating the steps, and sequentially coupling Fmoc-Glu-OtBu and HO-OC- (CH) ₂ ) ₁₆ -CO-OtBu, wherein the Fmoc-Glu-OtBu coupling dissolution and washing solvent after coupling is DMF. Coupling the target peptide sequence, washing with DCM for 6 times, and drying to obtain COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu, weighing 70.26g.

(2)COOtBu(CH ₂ ) ₁₆ Cleavage of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

70.26g of COOtBu (CH) was weighed out from the above-mentioned peptide resin ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu, added to 706.8ml of cracking reagent (TFE/DCM = 80/20) and cracked at 25 + -5 deg.CReacting for 1-4 h. After the reaction is finished, filtering, concentrating, washing with saturated salt water, extracting with DCM, combining organic phases, drying with anhydrous sodium sulfate, and concentrating to obtain COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu, weighing 36.20g.

(3) Synthesis of Fmoc-Ala-Lys-OH:

H-Lys (Boc) -OH (10.8 g, 1.1eq.) was dissolved in 600ml of THF as a mixed solvent: h ₂ O =1:1, completely dissolving and stirring, adjusting the pH value to 7.5-11.0 by using sodium hydroxide aqueous solution, adding Fmoc-Ala-OSu (17.1g, 1.05eq.), and adjusting the pH value to 7.5-11.0 after the addition. TLC monitors the reaction progress, after the reaction is finished, petroleum ether is used for extraction for 3 times, and the product is discarded. The aqueous layer was adjusted to pH 2-3 with dilute hydrochloric acid, the aqueous layer was extracted 3 times with DCM, 200 ml/time, and the organic phases were combined. 600ml of trifluoroacetic acid and 30ml of 3-mercaptopropionic acid are added and stirred for reaction for 2 to 4 hours. After the reaction is finished, concentrating under reduced pressure until no liquid drips, adding a mixed solvent (petroleum ether/isopropyl ether = 1) and pulping and washing for 1h, filtering, washing a filter cake by using petroleum ether, and drying a solid in vacuum to obtain Fmoc-Ala-Lys-OH with the weight of 14.65g.

(4)Fmoc-Ala-Lys(COOtBu(CH ₂ ) ₁₆ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -OH:

weighing COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu was dissolved in 600ml of DCM, and HOPfp (8.8g, 1.2eq.) was added thereto with stirring at room temperature and dissolved therein. DCC (9.9g, 1.2eq.) was weighed and dissolved in 400ml of DCM, and the solution was added dropwise to the reaction solution and stirred for 2-6 hours. TLC (thin layer chromatography) detection is carried out to ensure that the raw materials completely react, the reaction solution is filtered, white solid is filtered out, the white solid is washed twice by DCM (DCM), the filtrate is collected and concentrated under reduced pressure until no liquid drips, and rotary evaporation is stopped to obtain COOtBu (CH) ₂ ) ₁₆ Dissolving CO-Glu (AEEA-AEEA-OPfp) -OtBu in appropriate amount of acetonitrile, and filtering.

Fmoc-Ala-Lys-OH (14.65g, 1.1eq.) was dissolved in 800-1000ml of a mixed solvent (acetonitrile: water = 1), stirred at room temperature until the solution became clear, adjusted to pH with aqueous sodium hydroxide solution, and measured to pH 7.5-11.0. Stirring to separate COOtBu (CH) ₂ ) ₁₆ Slowly dripping CO-Glu (AEEA-AEEA-OPfp) -OtBu solution into the Fmoc-Ala-Lys-OH mixed solution, controlling the pH value to be 7.5-11.0, reacting for 2-8h at room temperature,TLC detected the starting material reaction was complete, filtered to remove a white solid, which was washed once with a mixed solvent (acetonitrile: water = 1), and the combined filtrates were collected and subjected to purification to give Fmoc-Ala-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH, 47.85g in weight.

Example 2

Fmoc-Lys(COOtBu(CH ₂ ) ₁₈ Preparation of CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH

Structural formula (xvi):

(1)COOtBu(CH ₂ ) ₁₈ synthesis of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

weighing 40.0g of CTC resin (40 mmol, substitution value of 1.10 mmol/g), and adding into a polypeptide synthesis reaction column; weighing 30.8g of Fmoc-AEEA-OH, adding 200ml of DCM for dissolving until the mixture is clear, then adding 25.8g of DIEA, and fully stirring; then adding the mixed solution into a reaction column, stirring and reacting for 2.0h, and discharging waste.

6.4g of methanol and 12.9g of DIEA were weighed, 320ml of DCM was added and stirred uniformly, and a solution of methanol/DIEA in DCM was added to the reaction column and stirred for reaction for 1.0h. Waste, DCM wash 4 times.

Repeating the steps to couple Fmoc-Glu-OtBu and HO-OC- (CH) in sequence ₂ ) ₁₈ -CO-OtBu, wherein the Fmoc-Glu-OtBu coupling dissolution and washing solvent after coupling is DMF. Coupling of the target peptide sequence was completed, washed 6 times with DCM and dried to yield 71.68g of COOtBu (CH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA-CTC resin) -OtBu.

(2)COOtBu(CH ₂ ) ₁₈ Cleavage of CO-Glu (AEEA-AEEA-CTC resin) -OtBuSolution:

71.68g of COOtBu (CH) as described above are weighed out ₂ ) ₁₈ CO-Glu (AEEA-AEEA-CTC resin) -OtBu was added to 706.8ml of a cracking reagent (TFE/DCM = 80/20) and subjected to a cracking reaction at 25. + -. 5 ℃ for 1 to 4 hours. After the reaction was complete, it was filtered and the resin was washed with DCM, 150 ml/time. Mixing the lysates, concentrating, washing with saturated saline, extracting with DCM, mixing the organic phases, drying with anhydrous sodium sulfate, and concentrating to obtain COOtBu (CH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA-OH) -OtBu, 35.86g in weight.

(3) Synthesis of Fmoc-Lys-Ala-OH:

H-Ala-OH (3.9 g, 1.1eq.) was dissolved in 600ml of THF as a mixed solvent: h ₂ O =1:1, dissolving and stirring completely, adjusting the pH value to 7.5-11.0 by using sodium hydroxide aqueous solution, adding Fmoc-Lys (Boc) -OSu (23.7g, 1.05eq.) and adjusting the pH value to 7.5-11.0 after the addition. TLC monitors the reaction progress, after the reaction is finished, petroleum ether is used for extraction for 3 times, and the product is discarded. Adjusting the pH of the water layer to 2-3 by using dilute hydrochloric acid, extracting the water layer for 3 times by using DCM, combining organic phases and concentrating. To the concentrate was added 100ml of lysate TFA/Mpr/H ₂ O =95/2.5/2.5 (v/v), stirring the reaction for 2-4h. After the reaction, isopropyl ether was settled, washed, filtered, and vacuum-dried to obtain Fmoc-Lys-Ala-OH with a weight of 14.90g.

(4)Fmoc-Lys(COOtBu(CH ₂ ) ₁₈ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH:

reacting COOtBu (CH) ₂ ) ₁₈ Dissolving CO-Glu (AEEA-AEEA) -OtBu) -OH in 800ml of DCM until the mixture is clear, adding HOBT (0.5g, 0.1eq.), EDC (8.1g, 1.05eq.) and Fmoc-Lys-Ala-OH (14.90g, 1.05eq.), stirring the mixture at room temperature until the mixture is clear, adjusting the pH value of triethylamine to 7.5-11.0, and stirring the mixture for reaction for 6-10 hours. TLC (thin layer chromatography) detection is carried out to ensure that the raw materials completely react, the pH value of reaction liquid is adjusted to 6.0-7.0 by dilute hydrochloric acid, an organic phase is washed three times by saturated saline solution, the organic phase is decompressed and concentrated until no liquid drips, and the organic phase is purified to obtain Fmoc-Lys (COOtBuCH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH, weight 47.85g.

Example 3

Boc-Ala-Lys(COOtBu(CH ₂ ) ₁₆ Preparation of CO-Glu (AEEA-AEEA) -OtBu) -OH

Structural formula (la):

(1)COOtBu(CH ₂ ) ₁₆ synthesis of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

weighing 40.0g of CTC resin (40 mmol, the substitution value is 1.10 mmol/g), and adding the CTC resin into a polypeptide synthesis reaction column; weighing 30.8g of Fmoc-AEEA-OH, adding 200ml of DCM for dissolving until the mixture is clear, then adding 25.8g of DIEA, and fully stirring; then the mixed solution is added into a reaction column, stirred and reacted for 2.0 hours, and waste is discharged.

The reaction column was charged with 20% of PIPE/DMF and the Fmoc protection was removed twice, with deprotection times of respectively 5min +15min and 6 DMF washes. Fmoc-AEEA-OH (23.1g, 1.5eq.) is weighed and dissolved in 100ml DCM, DIC (11.4g, 2.25eq.) is added at 0-10 ℃ for activation for 4min and added into a synthesis reaction column, and the mixture reacts for 2-4h under the protection of nitrogen at 25 +/-5 ℃. Waste, DCM wash 4 times.

Repeating the steps, and sequentially coupling Fmoc-Glu-OtBu and HO-OC- (CH) ₂ ) ₁₆ -CO-OtBu, wherein the solvent for Fmoc-Glu-OtBu coupling dissolution and washing after coupling is DMF. Coupling the target peptide sequence, washing 6 times with DCM, and drying to obtain COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu, weighing 71.08g.

(2)COOtBu(CH ₂ ) ₁₆ Cleavage of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

71.08g of COOtBu (CH) in the above-mentioned peptide resin was weighed ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu was added to 706.8ml of a cracking reagent (TFE/DCM = 80/20) and subjected to a cracking reaction at 25. + -. 5 ℃ for 1 to 4 hours. After the reaction is finished, filtering, concentrating, washing with saturated salt water, extracting with DCM, combining organic phases, drying with anhydrous sodium sulfate, and concentrating to obtain COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu, weighing 36.20g.

(3) Synthesis of Boc-Ala-Lys-OH:

H-Lys-OH. HCl (8.0 g, 1.1eq.) was purified with 600ml of THF, a mixed solvent: h ₂ O =1:1, completely dissolving and stirring, adjusting the pH value of a sodium hydroxide aqueous solution to 7.5-11.0, adding Boc-Ala-OSu (17.1g, 1.05eq.) and adjusting the pH value to 7.5-11.0 after the addition. The progress of the reaction was monitored by TLC, and after completion of the reaction, it was extracted 3 times with petroleum ether and discarded. Adjusting the pH of a water layer to 2-3 by using dilute hydrochloric acid, extracting the water layer by using ethyl acetate for 3 times, combining organic phases, washing by using saturated saline solution, drying by using anhydrous sodium sulfate, concentrating until no liquid drops, adding a mixed solvent (petroleum ether/isopropyl ether = 1), pulping and washing for 1h, filtering, washing a filter cake by using petroleum ether, and drying the solid in vacuum to obtain Boc-Ala-Lys-OH solid with the weight of 11.80g.

(4)Boc-Ala-Lys(COOtBu(CH ₂ ) ₁₆ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -OH:

weighing COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu in 600ml DCM was added HOPfp (8.8g, 1.2eq.) with stirring at room temperature and dissolved to dryness with stirring. DCC (9.9g, 1.2eq.) was weighed and dissolved in 400ml of DCM, added dropwise to the reaction solution, and reacted for 2-6 hours with stirring. TLC (thin-layer chromatography) for detecting complete reaction of raw materials, filtering reaction liquid, filtering out white solid, washing the white solid twice with DCM (DCM), collecting filtrate, concentrating under reduced pressure until no liquid drips, stopping rotary evaporation to obtain COOtBu (CH) ₂ ) ₁₆ Dissolving CO-Glu (AEEA-AEEA-OPfp) -OtBu in appropriate amount of acetonitrile, and filtering.

Boc-Ala-Lys-OH (11.80g, 1.1eq.) was dissolved in 800-1000ml of a mixed solvent (acetonitrile: water = 1), stirred at room temperature until the solution became clear, adjusted to pH with aqueous sodium hydroxide solution, and measured as pH 7.5-11.0. Stirring to separate COOtBu (CH) ₂ ) ₁₆ Slowly dropwise adding a CO-Glu (AEEA-AEEA-OPfp) -OtBu solution into a Boc-Ala-Lys-OH mixed solution, controlling the pH value to be 7.5-11.0, reacting for 2-8h at room temperature, detecting that raw materials are completely reacted by TLC, filtering, removing a white solid, washing the white solid once by using a mixed solvent (acetonitrile: water = 1) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH, 40.02g in weight.

Example 4

Boc-Lys(COOtBu(CH ₂ ) ₁₆ Preparation of CO-Glu (AEEA-AEEA) -OtBu) -Ala-OAll

Structural formula (xvi):

(1)COOtBu(CH ₂ ) ₁₆ synthesis of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

weighing 40.0g of CTC resin (40 mmol, substitution value of 1.10 mmol/g), and adding into a polypeptide synthesis reaction column; weighing 30.8g of Fmoc-AEEA-OH, adding 200ml of DCM for dissolving until the mixture is clear, then adding 25.8g of DIEA, and fully stirring; then the mixed solution is added into a reaction column, stirred and reacted for 2.0 hours, and waste is discharged.

Repeating the steps, and sequentially coupling Fmoc-Glu-OtBu and HO-OC- (CH) ₂ ) ₁₆ -CO-OtBu, wherein the Fmoc-Glu-OtBu coupling dissolution and washing solvent after coupling is DMF. Coupling of the target peptide sequence was completed, washed 6 times with DCM and dried to give 69.86g of COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu.

(2)COOtBu(CH ₂ ) ₁₆ Cleavage of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

69.86g of the above-mentioned COOtBu (CH) was weighed out ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu was added to 706.8ml of a cracking reagent (TFE/DCM = 80/20) and subjected to a cracking reaction at 25. + -. 5 ℃ for 1 to 4 hours. After the reaction was complete, it was filtered and the resin was washed with DCM, 150 ml/time. Mixing the lysates, concentrating,Washing with saturated saline, extracting with DCM, mixing organic phases, drying with anhydrous sodium sulfate, and concentrating to obtain COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu, 35.22g in weight.

(3)Boc-Lys(COOtBu(CH ₂ ) ₁₆ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -OH:

weighing COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu in 600ml DCM was added HOPfp (8.8g, 1.2eq.) with stirring at room temperature and dissolved to dryness with stirring. DCC (9.9g, 1.2eq.) was weighed and dissolved in 400ml of DCM, and added dropwise to the reaction solution, and the reaction was stirred for 2 hours. TLC detection of complete reaction of raw materials, filtration, washing of white solid twice with DCM, collection of filtrate, and concentration under reduced pressure to obtain COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OPfp) -OtBu was dissolved in an appropriate amount of acetonitrile, and a white solid was precipitated and removed by filtration.

Boc-Lys-OH (10.8g, 1.1eq.) was weighed and dissolved in a mixed solvent of 800 (acetonitrile: water = 1. Slowly dropwise adding COOtBu (CH) under stirring ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OPfp) -OtBu solution, pH value is controlled to be 7.5-11.0, the reaction is carried out for 2-8h at room temperature, TLC detection raw material reaction is complete, filtration is carried out to remove white solid, the white solid is washed once by mixed solvent (acetonitrile: water = 1), and the combined filtrates are collected and sent to purification treatment to obtain Boc-Lys (COOtBu (CH ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH, weighing 36.6g.

(4)Boc-Lys(COOtBu(CH ₂ ) ₁₆ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -Ala-OAll:

Boc-Lys (COOtBu (CH) ₂ ) ₁₆ Adding CO-Glu (AEEA-AEEA) -OtBu) -OH into 800ml of ACN to dissolve until the solution is clear; CDI (13.0 g,2.0 eq.) was weighed and dissolved in 400ml of DCM, and added dropwise to the above reaction solution, followed by reaction at 35. + -. 5 ℃ for 1-2 hours with stirring. TLC detects that the raw material is completely reacted and is cooled to 20 +/-5 ℃.

H-Ala-OAll · HCl (7.3g, 1.1eq.) was weighed and dissolved in 800-1000ml of a mixed solvent (acetonitrile: water = 1), stirred at room temperature until clear, and pH was adjusted to 7.0-9.0 with 25% aqueous ammonia. Slowly dropwise adding the mixture into the reaction solution, and controlling the reactionThe temperature is less than or equal to 22 ℃, the reaction is carried out for 1-2h, and the intermediate is detected by TLC to be completely reacted. Concentrating under reduced pressure to remove acetonitrile, adding water, continuously distilling to remove acetonitrile, extracting with ethyl acetate twice, mixing organic phases, washing with water, standing for layering, separating water layer, and concentrating organic phase to obtain Boc-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OAll, purified and weighed 41.03g.

Example 5

Dde-Ala-Lys(COOBzl(CH ₂ ) ₁₆ Preparation of CO-Glu (AEEA-AEEA) -OtBu) -OH

Structural formula (xvi):

(1)COOBzl(CH ₂ ) ₁₆ synthesis of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

weighing 40.0g of CTC resin (40 mmol, substitution value of 1.10 mmol/g), and adding into a polypeptide synthesis reaction column; weighing 30.8g of Fmoc-AEEA-OH, adding 200ml of DCM to dissolve until the mixture is clear, then adding 25.8g of DIEA, and fully stirring; then adding the mixed solution into a reaction column, stirring and reacting for 2.0h, and discharging waste.

The reaction column was charged with 20% of PIPE/DMF and the Fmoc protection was removed twice, with deprotection times of respectively 5min +15min and 6 DMF washes. Weighing Fmoc-AEEA-OH (23.1g, 1.5eq.) and dissolving in 100ml DCM, adding DIC (11.4g, 2.25eq.) and activating at 0-10 ℃ for 4min, adding into a synthesis reaction column, and reacting for 2-4h at 25 +/-5 ℃ under the protection of nitrogen. Waste, DCM wash 4 times.

Repeating the steps, and sequentially coupling Fmoc-Glu-OtBu and HO-OC- (CH) ₂ ) ₁₆ -CO-OBzl, wherein the Fmoc-Glu-OtBu coupling dissolution and washing solvent after coupling is DMF. Coupling the target peptide sequence, washing 6 times with DCM, and drying to obtain COOBzl (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu, weighing 68.95g.

(2)COOBzl(CH ₂ ) ₁₆ Cleavage of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

68.95g of COOBzl (CH) are weighed out ₂ ) ₁₆ CO-Glu (AEEA-AEEA-CTC resin) -OtBu was added to 706.8ml of a cracking reagent (TFE/DCM = 80/20) and subjected to a cracking reaction at 25. + -. 5 ℃ for 1 to 4 hours. After the reaction, filtering, concentrating, washing with saturated salt water, extracting with DCM, combining organic phases, drying with anhydrous sodium sulfate, and concentrating to obtain COOBzl (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu, weighing 34.62g.

(3) Synthesis of Dde-Ala-Lys-OH:

H-Lys-OH & HCl (8.0 g, 1.1eq.) was purified using 600ml of mixed solvent THF: h ₂ O =1:1, dissolving and stirring completely, adjusting the pH value of the sodium hydroxide aqueous solution to 7.5-11.0, and adding Dde-Ala-OSu

(14.7g, 1.05eq.) and after addition the pH is adjusted to 7.5-11.0. The progress of the reaction was monitored by TLC, and after completion of the reaction, it was extracted 3 times with petroleum ether and discarded. Adjusting the pH of a water layer to 2-3 by using dilute hydrochloric acid, extracting the water layer by using ethyl acetate for 3 times, combining organic phases, washing by using saturated saline solution, drying by using anhydrous sodium sulfate, concentrating until no liquid drops, adding a mixed solvent (petroleum ether/isopropyl ether = 1) and pulping and washing for 1h, filtering, washing a filter cake by using petroleum ether, and drying a solid in vacuum to obtain Dde-Ala-Lys-OH with the weight of 12.70g.

(4)Dde-Ala-Lys(COOBzl(CH ₂ ) ₁₆ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -OH:

weighing COOBzl (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA-OH) -OtBu was added to 600ml of DCM with stirring at room temperature, HOBT (0.5g, 0.1eq.), EDC (8.1g, 1.05eq.) and Dde-Ala-Lys-OH (12.70g, 1.1eq.) were added to the mixture with stirring at room temperature, the mixture was stirred to a clear solution at room temperature, triethylamine was added to adjust the pH to 7.5-11.0, and the mixture was reacted with stirring for 6-10 hours. TLC detecting the reaction of raw material completely, adjusting pH of reaction solution to 6.0-7.0 with dilute hydrochloric acid, washing with saturated saline solution for three times, concentrating under reduced pressure until no liquid drips, and purifying to obtain Dde-Ala-Lys (COOBzl (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH, weighing 46.97g.

Example 6

Alloc-Lys(COOtBu(CH ₂ ) ₁₈ Preparation of CO-Glu (AEEA-AEEA) -OMe) -Ala-OH formula:

(1)Alloc-Lys(COOtBu(CH ₂ ) ₁₈ synthesis of CO-Glu (AEEA-AEEA) -OMe) -Aal-CTC resin:

weighing 40.0g of CTC resin (40 mmol, the substitution value is 1.10 mmol/g), and adding the CTC resin into a polypeptide synthesis reaction column; weighing 24.9g of Fmoc-Ala-OH, adding 200ml of DMF, dissolving until the mixture is clear, then adding 25.8g of DIEA, and fully stirring; then adding the mixed solution into a reaction column, stirring and reacting for 2.0h, and discharging waste.

6.4g of methanol and 12.9g of DIEA were weighed, 320ml of DMF was added and stirred uniformly, and then a DMF solution of methanol/DIEA was added to the reaction column and stirred for reaction for 1.0 hour. The waste was discarded and washed 4 times with DMF.

The reaction column was charged with 20% PIPE/DMF and the Fmoc protection was removed twice, the deprotection times were respectively 5min +15min and DMF washes 6 times. Alloc-Lys (Fmoc) -OH (27.2g, 1.5eq.) is weighed and dissolved in 100ml DMF, DIC (11.4g, 2.25eq.) is added at the temperature of 0-10 ℃ for activation for 4min and then added into a synthetic reaction column for reaction for 2-4h under the protection of nitrogen at the temperature of 25 +/-5 ℃. Waste, DCM wash 4 times.

Adding 2% hydrazine hydrate/DMF to the reaction column to remove Fmoc protection three times, wherein the deprotection time is 10min +30min respectively, and washing 6 times with DMF. Weighing Fmoc-AEEA-OH (23.1g, 1.5eq.) and dissolving in 100ml DCM, adding DIC (11.4g, 2.25eq.) and activating at 0-10 ℃ for 4min, adding into a synthesis reaction column, and reacting for 2-4h at 25 +/-5 ℃ under the protection of nitrogen. Waste, DCM wash 4 times.

Repeating the steps, and sequentially coupling Fmoc-Glu-OMe and HO-OC- (CH) ₂ ) ₁₈ -CO-OtBu，Wherein the washing solvent after Fmoc-Glu-OMe coupling dissolution and coupling is DMF. Coupling of the target peptide sequence was completed, washed 6 times with DCM and dried to yield Alloc-Lys (COOtBu (CH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OMe) -Aal-CTC resin, 76.85g in weight.

(2)Alloc-Lys(COOtBu(CH ₂ ) ₁₈ Cleavage of CO-Glu (AEEA-AEEA) -Aal-CTC resin-OMe:

76.85g of Alloc-Lys (COOtBu (CH)) as the above-mentioned peptide resin was weighed ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OMe) -Aal-CTC resin is added into 706.8ml of cracking reagent (TFE/DCM = 80/20), and the cracking reaction is carried out for 1-4 h at 25 +/-5 ℃. After the reaction is finished, filtering, concentrating, washing with saturated salt water, extracting with DCM, combining organic phases, drying with anhydrous sodium sulfate, concentrating, and purifying to obtain Alloc-Lys (COOtBu (CH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OMe) -Ala-OH, weighing 35.87g.

Example 7

H-Ala-Lys(COOH(CH ₂ ) ₁₆ CO-Glu(AEEA-AEEA)-OH)-OH

Structural formula (la):

following the procedure of example 3, boc-Ala-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH with a weight of 40.85g;

Boc-Ala-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH was added to 800-1000ml of a mixed solvent (acetonitrile: water = 1), stirring to clear at room temperature, heating to 40 +/-5 ℃, introducing hydrochloric acid gas into the solution, reacting for 6-10h, monitoring the reaction process by TLC, concentrating under reduced pressure to remove acetonitrile after the reaction is finished, adding water, continuously distilling to remove acetonitrile, extracting with ethyl acetate twice, combining organic phases, washing with water, standing for layering, separating out a water layer, and concentrating the organic phase to obtain H-Ala-Lys (COOH (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OH) -OH, purified to give a solid weighing 37.65g.

Example 8

H-Lys(COOH(CH ₂ ) ₁₆ CO-Glu(AEEA-AEEA)-OH)-Ala-OH

Structural formula (la):

the procedure of example 4, in which H-Ala-OH (3.9 g, 1.1eq.) was used as a starting material for liquid phase synthesis, yielded Boc-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH, weight 41.03g;

Boc-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH was added to 800-1000ml of a mixed solvent (acetonitrile: water = 1), stirring to clear at room temperature, heating to 40 +/-5 ℃, introducing hydrochloric acid gas into the solution, reacting for 6-10h, monitoring the reaction process by TLC, concentrating under reduced pressure to remove acetonitrile after the reaction is finished, adding water, continuously distilling to remove acetonitrile, extracting with ethyl acetate twice, combining organic phases, washing with water, standing for layering, separating out a water layer, and concentrating the organic phase to obtain H-Lys (COOH (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OH) -Ala-OH was purified to give a solid weighing 37.65g.

Example 9

Fmoc-Ala-Lys(COOH(CH ₂ ) ₁₀ Preparation of CO-Glu (AEEA-AEEA) -OtBu) -OH

Structural formula (xvi):

(1)COOBzl(CH ₂ ) ₁₀ synthesis of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

The reaction column was charged with 20% PIPE/DMF and the Fmoc protection was removed twice, the deprotection times were respectively 5min +15min and DMF washes 6 times. Fmoc-AEEA-OH (23.1g, 1.5eq.) is weighed and dissolved in 100ml DCM, DIC (11.4g, 2.25eq.) is added at 0-10 ℃ for activation for 4min and added into a synthesis reaction column, and the mixture reacts for 2-4h under the protection of nitrogen at 25 +/-5 ℃. Waste, DCM wash 4 times.

Repeating the steps, and sequentially coupling Fmoc-Glu-OtBu and HO-OC- (CH) ₂ ) ₁₀ -CO-Bzl, wherein DMF is used as a solvent for Fmoc-Glu-OtBu coupling dissolution and washing after coupling. Coupling of the target peptide sequence was completed, washed 6 times with DCM, and dried to give COOBzl (CH) ₂ ) ₁₀ CO-Glu (AEEA-AEEA-CTC resin) -OtBu, weighing 66.10g.

(2)COOBzl(CH ₂ ) ₁₀ Cleavage of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

66.10g of the above peptide resin COOBzl (CH) was weighed ₂ ) ₁₀ CO-Glu (AEEA-AEEA-CTC resin) -OtBu was added to 706.8ml of a cracking reagent (TFE/DCM = 80/20) and subjected to a cracking reaction at 25. + -. 5 ℃ for 1 to 4 hours. After the reaction, filtering, concentrating, washing with saturated salt water, extracting with DCM, combining organic phases, drying with anhydrous sodium sulfate, and concentrating to obtain COOBzl (CH) ₂ ) ₁₀ CO-Glu (AEEA-AEEA-OH) -OtBu, weighing 33.93g.

(3) Synthesis of Fmoc-Ala-Lys-OH:

H-Lys-OH & HCl (8.0 g, 1.1eq.) was purified using 600ml of mixed solvent THF: h ₂ O =1:1, completely dissolving and stirring, adjusting the pH value to 7.5-11.0 by using sodium hydroxide aqueous solution, adding Fmoc-Ala-OSu (17.1g, 1.05eq.), and adjusting the pH value to 7.5-11.0 after the addition. TLC monitors the reaction progress, after the reaction is finished, petroleum ether is used for extraction for 3 times, and the product is discarded. Adjusting the pH of a water layer to 2-3 by using dilute hydrochloric acid, extracting the water layer by using ethyl acetate for 3 times, combining organic phases, washing by using saturated saline solution, drying by using anhydrous sodium sulfate, concentrating until no liquid drops, adding a mixed solvent (petroleum ether/isopropyl ether = 1).

(4)Fmoc-Ala-Lys(COOBzl(CH ₂ ) ₁₀ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -OH:

weighing COOBzl (CH) ₂ ) ₁₀ CO-Glu (AEEA-AEEA-OH) -OtBu in 600ml DCM was added HOPfp (8.8g, 1.2eq.) with stirring at room temperature and dissolved to dryness with stirring. DCC (9.9g, 1.2eq.) was weighed and dissolved in 400ml of DCM, added dropwise to the reaction solution, and reacted for 2-6 hours with stirring. TLC detects that the raw material completely reacts, the reaction liquid is filtered, white solid is filtered out, the white solid is washed twice by DCM, filtrate is collected, reduced pressure concentration is carried out until no liquid drips, rotary evaporation is stopped, and COOBzl (CH) is obtained ₂ ) ₁₀ Dissolving CO-Glu (AEEA-AEEA-OPfp) -OtBu in appropriate amount of acetonitrile, and filtering.

Fmoc-Ala-Lys-OH (14.00g, 1.1eq.) was dissolved in 800-1000ml of a mixed solvent (acetonitrile: water = 1), stirred at room temperature until the solution became clear, adjusted to pH with aqueous sodium hydroxide solution, and measured to pH 7.5-11.0. COOBzl (CH) is added under stirring ₂ ) ₁₀ Slowly dropwise adding a CO-Glu (AEEA-AEEA-OPfp) -OtBu solution into a mixed solution of Fmoc-Ala-Lys-OH, controlling the pH value to be 7.5-11.0, reacting at room temperature for 2-8h, detecting the complete reaction of raw materials by TLC, filtering, removing a white solid, washing the white solid once by using a mixed solvent (acetonitrile: water = 1), collecting the combined filtrate, and concentrating under reduced pressure to obtain Fmoc-Ala-Lys (COOBzl (CH ₂ ) ₁₀ CO-Glu (AEEA-AEEA) -OtBu) -OH, weighing 46.82g.

(5)Fmoc-Ala-Lys(COOOH(CH ₂ ) ₁₀ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -OH:

Fmoc-Ala-Lys (COOBzl (CH) was weighed ₂ ) ₁₀ CO-Glu (AEEA-AEEA) -OtBu) -OH 46.82g and 10% Pd-C5.0 g were added to 800ml of tetrahydrofuran, the mixture was purged with nitrogen, and H was purged ₂ The reaction pressure was controlled at 3bar. Stirring and reacting for 8-16h. After the reaction, the reaction solution was concentrated under reduced pressure after nitrogen substitution. 300ml of H are added ₂ And O, adjusting the pH value of the reaction solution to 7.5-11.0 by using NaOH solution, extracting by using petroleum ether for three times, and discarding an organic phase. Adjusting pH of the aqueous phase to 6.0-7.0 with dilute hydrochloric acid, extracting with ethyl acetate, drying with anhydrous sodium sulfate, concentrating under reduced pressure, and purifying to obtain Fmoc-Ala-Lys (COOH (CH) ₂ ) ₁₀ CO-Glu(AEEA-AEEA)-OtBu)-OH, weight 45.25g.

Example 10

Fmoc-Lys(COOH(CH ₂ ) ₂₀ Preparation of CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH

Structural formula (xvi):

(1)COOBzl(CH ₂ ) ₂₀ synthesis of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

Repeating the steps, and sequentially coupling Fmoc-Glu-OtBu and HO-OC- (CH) ₂ ) ₂₀ -CO-Bzl, wherein DMF is used as a solvent for Fmoc-Glu-OtBu coupling dissolution and washing after coupling. Coupling of the target peptide sequence was completed, washed 6 times with DCM, and dried to give 73.05g of COOBzl (CH) ₂ ) ₂₀ CO-Glu (AEEA-AEEA-CTC resin) -OtBu.

(2)COOBzl(CH ₂ ) ₂₀ Cleavage of CO-Glu (AEEA-AEEA-CTC resin) -OtBu:

73.05g of the above COOBzl (CH) are weighed out ₂ ) ₂₀ CO-Glu (AEEA-AEEA-CTC resin) -OtBu was added to 706.8ml of a cracking reagent (TFE/DCM = 80/20) and subjected to a cracking reaction at 25. + -. 5 ℃ for 1 to 4 hours. After the reaction is finished, filtering, useThe resin was washed with DCM, 150 ml/time. Mixing the lysates, concentrating, washing with saturated saline, extracting with DCM, mixing the organic phases, drying with anhydrous sodium sulfate, and concentrating to obtain COOBzl (CH) ₂ ) ₂₀ CO-Glu (AEEA-AEEA-OH) -OtBu, weighing 37.66g.

(3) Synthesis of Fmoc-Lys-Ala-OH:

H-Ala-OH (3.9g, 1.1eq.) was dissolved in 600ml of THF: h ₂ O =1:1, dissolving and stirring completely, adjusting the pH value to 7.5-11.0 by using sodium hydroxide aqueous solution, adding Fmoc-Lys (Boc) -OSu (23.7g, 1.05eq.) and adjusting the pH value to 7.5-11.0 after the addition. The progress of the reaction was monitored by TLC, and after completion of the reaction, it was extracted 3 times with petroleum ether and discarded. The aqueous layer was adjusted to pH 2-3 with dilute hydrochloric acid, the aqueous layer was extracted 3 times with DCM, 200 ml/time, and the organic phases were combined. 600ml of trifluoroacetic acid and 30ml of 3-mercaptopropionic acid are added and stirred for reaction for 2 to 4 hours. After the reaction is finished, concentrating under reduced pressure until no liquid drips, adding a mixed solvent (petroleum ether/isopropyl ether = 1) and pulping and washing for 1h, filtering, washing a filter cake by using petroleum ether, and drying the solid in vacuum to obtain Fmoc-Lys-Ala-OH with the weight of 14.90g.

(4)Fmoc-Lys(COOBzl(CH ₂ ) ₂₀ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH:

reacting COOBzl (CH) ₂ ) ₂₀ Dissolving CO-Glu (AEEA-AEEA) -OtBu) -OH in 800ml of DCM until the mixture is clear, adding HOBT (0.5g, 0.1eq.), EDC (8.1g, 1.05eq.) and Fmoc-Lys-Ala-OH (14.90g, 1.05eq.), stirring the mixture at room temperature until the mixture is clear, adjusting the pH value of triethylamine to 7.5-11.0, and stirring the mixture for reaction for 6-10 hours. TLC detection of the reaction completion of the raw materials, adjusting the pH value of the reaction solution to 6.0-7.0 with dilute hydrochloric acid, washing the organic phase with saturated saline solution for three times, and concentrating under reduced pressure to obtain Fmoc-Lys (COOBzlCH) ₂ ) ₂₀ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH, weighing 50.06g.

(5)Fmoc-Lys(COOOH(CH ₂ ) ₂₀ Synthesis of CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH:

Fmoc-Lys (COOBzlCH) was weighed ₂ ) ₂₀ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH 50.06g, 10% Pd-C5.0 g, added to 800ml tetrahydrofuran, nitrogen replaced, H purged ₂ The reaction pressure was controlled at 3bar. Stirring and reacting for 8-16h. After the reaction, nitrogen gas was replacedThen, the reaction mixture was concentrated under reduced pressure. 300ml of H are added ₂ And O, adjusting the pH value of the reaction solution to 7.5-11.0 by using NaOH solution, extracting by using petroleum ether for three times, and discarding an organic phase. Adjusting pH of the water phase to 6.0-7.0 with dilute hydrochloric acid, extracting with ethyl acetate, drying with anhydrous sodium sulfate, concentrating, and purifying to obtain Fmoc-Lys (COOBzlCH) ₂ ) ₂₀ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH, weighing 48.42g.

Example 11

Semaglutide synthesis:

(1) Synthesis of Semaglutide peptide resin A

50g of Wang resin (20 mmol, substitution value 0.40 mmol/g) was weighed, added to a polypeptide synthesis reaction column, added with 500ml of DMF, swollen for 30min, and drained. Washed twice with DMF and then drained. Fmoc-Gly-OH (11.90g, 2.0eq.) and HOBt (5.90g, 2.2eq.) and DMAP (0.50g, 0.2eq.) are weighed and dissolved in 200ml of DMF, DIC (7.60g, 3.0eq.) is added at the temperature of 0-10 ℃ for activation for 4min, the mixture is added into a synthesis reaction column, and the reaction is carried out for 2-4h at the temperature of 25 +/-5 ℃ under the protection of nitrogen. It was drained and washed 4 times with DMF.

Add 20% of PIPE/DMF to remove Fmoc protection twice, deprotection times were 5min +15min, respectively, DMF washes 6 times. Add 20% of PIPE/DMF to remove Fmoc protection twice, deprotection times are respectively 5min +15min and DMF washes 6 times. Fmoc-Arg (Pbf) -OH (38.93g, 4.0eq.) and HOBt (11.89g, 3.3eq.) are weighed and dissolved in 200ml of DMF, DIC (15.15g, 4.5eq.) is added at the temperature of 0-10 ℃ for activation for 4min, the mixture is added into a synthesis reaction column, and the reaction is carried out for 2-4h under the protection of nitrogen at the temperature of 25 +/-5 ℃. It was drained and washed 4 times with DMF.

Repeating the above steps, coupling Fmoc-Gly-OH, fmoc-Arg (Pbf) -OH, fmoc-Val-OH, fmoc-Leu-OH, fmoc-Trp (Boc) -OH, fmoc-Ala-OH, fmoc-Ile-OH, fmoc-Phe-OH, fmoc-Glu (OtBu) -OH, fmoc-Ala-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH, fmoc-Ala-OH, fmoc-Gln (Trt) -OH, fmoc-Gly-OH, fmoc-Glu (OtBu) -OH, fmoc-Leu-OH, fmoc-Tyr (tBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Val-OH, fmoc-Asp (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Thr (tBu) -OH, fmoc-His (Trt) -b-Glu (OtBu) -Gly-Thr (tBu) -Phe-OH. Fmoc protecting group is removed, and then washed 4 times with DMF, 3 times with DCM, meOHWashing for 3 times, and drying to obtain the Semaglutide resin A: H-His (Trt) -Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Val-Ser (tBu) -Ser (tBu) -Tyr (tBu) -Leu-Glu (OtBu) -Gly-Gln (Trt) -Ala-Ala-Lys (COOtBu (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu-Glu (OtBu) -Phe-Ile-Ala-Trp (Boc) -Leu-Val-Arg (Pbf) -Gly-Arg (Pbf) -Gly-Wang resin, weight 188.23g.

(2) Semaglutide peptide resin A cleavage

188.23g of the Semaglutide peptide resin A is weighed and then 1882.3ml of cracking reagent TFA/Tis/H is added ₂ O/Mpr/m-Cresol =92.5/0.5/1/2/4, and the cracking reaction is carried out for 1-4 h at 25 +/-5 ℃. After the reaction was complete, filtration was carried out and the resin was washed with 200ml TFA. Adding the filtrate into 20L of frozen isopropyl ether for sedimentation, separating out solids, centrifuging, and washing to obtain a crude product of Semaglutide: H-His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys (HO-OC (CH) ₂ ) ₁₆ CO-gamma-Glu-AEEA-AEEA) -Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH, the weight is 86.43g, the yield is 95.1%, the purity is 76.9%, and the maximum single impurity is less than 5%.

(3) Refining of crude Semaglutide

Dissolving the crude Semaglutide in 10% acetonitrile water solution, filtering with 0.45 μm organic membrane, purifying and separating with reversed phase liquid chromatography, wherein the filler is reversed phase C18, particle diameter is 8 μm, and pore diameter is

Semaglutide depsipeptide was obtained by gradient elution using 0.1% aqueous TFA and acetonitrile, and had a weight of 26.95g, a yield of 31.6%, a purity of 99%, and a maximum single impurity of less than 0.2%.

Example 12

Semaglutide synthesis:

(1) Synthesis of Semaglutide peptide resin B

50g of Wang resin (20 mmol, substitution value 0.40 mmol/g) was weighed, added to a polypeptide synthesis reaction column, added with 500ml of DMF, swollen for 30min, and drained. Washed twice with DMF and then drained. Fmoc-Gly-OH (11.90g, 2.0eq.) and HOBt (5.90g, 2.2eq.) and DMAP (0.50g, 0.2eq.) are weighed and dissolved in 200ml of DMF, DIC (7.60g, 3.0eq.) is added at the temperature of 0-10 ℃ for activation for 4min, and the mixture is added into a synthesis reaction column and reacts for 2-4h under the protection of nitrogen at the temperature of 25 +/-5 ℃. It was drained and washed 4 times with DMF.

Add 20% of PIPE/DMF to remove Fmoc protection twice, deprotection times are respectively 5min +15min and DMF washes 6 times. Fmoc-Arg (Pbf) -OH (38.93g, 4.0eq.) and HOBt (11.89g, 3.3eq.) are weighed and dissolved in 200ml of DMF, DIC (15.15g, 4.5eq.) is added at the temperature of 0-10 ℃ for activation for 4min, the mixture is added into a synthesis reaction column, and the reaction is carried out for 2-4h under the protection of nitrogen at the temperature of 25 +/-5 ℃. It was drained and washed 4 times with DMF.

Repeating the above steps, coupling Fmoc-Gly-OH, fmoc-Arg (Pbf) -OH, fmoc-Val-OH, fmoc-Leu-OH, fmoc-Trp (Boc) -OH, fmoc-Ala-OH, fmoc-Ile-OH, fmoc-Phe-OH, fmoc-Glu (OtBu) -OH, dde-Ala-Lys (COOBzl (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu) -OH, coupling is finished and DMF washing is carried out for 4 times;

adding 2% hydrazine hydrate/DMF to remove Dde protection for three times, the deprotection time is 10min +30min respectively, and DMF washing for 6 times. Repeating amino acid coupling and Fmoc protecting group removal, and sequentially continuing coupling Fmoc-Ala-OH, fmoc-Gln (Trt) -OH, fmoc-Gly-OH, fmoc-Glu (OtBu) -OH, fmoc-Leu-OH, fmoc-Tyr (tBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Val-OH, fmoc-Asp (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Thr (tBu) -OH, fmoc-His (Trt) -Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-OH. Removing Fmoc protecting group, washing with DMF for 4 times, washing with DCM for 3 times, washing with MeOH for 3 times, and drying to obtain Semaglutide resin B: H-His (Trt) -Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Val-Ser (tBu) -Ser (tBu) -Tyr (tBu) -Leu-Glu (OtBu) -Gly-Gln (Trt) -Ala-Ala-Lys (COOBzl (CH) ₂ ) ₁₆ CO-Glu (AEEA-AEEA) -OtBu-Glu (OtBu) -Phe-Ile-Ala-Trp (Boc) -Leu-Val-Arg (Pbf) -Gly-Arg (Pbf) -Gly-Wang resin, 192.28g in weight.

(2) Semaglutide peptide resin cleavage

192.28g of the Semaglutide peptide resin B is weighed and added with 1922.8ml of a cleavage reagent TFA/Tis/H ₂ O/Mpr/m-Cresol =92.5/0.5/1/2/4, and the cracking reaction is carried out for 1-4 h at 25 +/-5 ℃. After the reaction was complete, filtration was carried out and the resin was washed with 200ml TFA. Adding the filtrate into 20L of frozen isopropyl ether, precipitating, and separating out solidCentrifuging, washing to obtain a crude product of Semaglutide with Bzl protecting group: H-His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys (BzlO-OC (CH) ₂ ) ₁₆ CO-gamma-Glu-AEEA-AEEA) -Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH, the weight was 86.43g.

(3) Semaglutide hydrogenation reduction

Weighing 86.43g of crude Semaglutide product with Bzl protecting group and 5.0g of 10% Pd-C, adding into 800ml of methanol, replacing with nitrogen, introducing H ₂ The reaction pressure was controlled at 3bar. Stirring and reacting for 8-16h. After the reaction, the reaction solution was concentrated to remove methanol by nitrogen substitution. 300ml of H are added ₂ And O, adjusting the pH value of the reaction solution to 7.5-11.0 by using NaOH solution, extracting by using petroleum ether for three times, and discarding an organic phase. Adjusting the pH value of the reaction solution to 6.0-7.0 by using dilute hydrochloric acid in the water phase, extracting by using ethyl acetate, drying by using anhydrous sodium sulfate, and concentrating to obtain a crude product of Semaglutide: H-His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys (HO-OC (CH) ₂ ) ₁₆ CO-gamma-Glu-AEEA-AEEA) -Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH, weight 81.5g, yield 96.8%, purity 75.8%, maximum single impurity less than 5%.

(4) Refining of crude Semaglutide

Semaglutide depsipeptide was obtained by gradient elution using 0.1% aqueous TFA and acetonitrile, in a weight of 25.80g, in a yield of 30.9%, in a purity of 99%, and in a maximum single impurity content of less than 0.2%.

Example 13

Tirzepatide synthesis:

(1) Synthesis of Tirzepatide peptide resin A

62.5g Rink Amide MBHA resin (20 mmol, substitution value of 0.32 mmol/g) is weighed into a polypeptide synthesis reaction column, 200ml DMF is added for swelling for 30min, and the mixture is drained. Washed twice with DMF and then drained.

Add 20% of PIPE/DMF to remove Fmoc protection twice, deprotection times are respectively 5min +15min and DMF washes 6 times. Fmoc-Ser (tBu) -OH (30.68g, 4.0eq.) and HOBt (11.89g, 4.4q.) are weighed and dissolved in 100ml of DMF, DIC (15.15ml, 6.0eq.) is added at the temperature of 0-10 ℃ for activation for 4min, the mixture is added into a synthesis reaction column, and the reaction is carried out for 2-4h under the protection of nitrogen at the temperature of 25 +/-5 ℃. It was drained and washed 4 times with DMF.

Repeating the above steps, coupling Fmoc-Pro-OH, fmoc-Ala-OH, fmoc-Gly-OH, fmoc-Ser (tBu) -OH, fmoc-Pro-OH, fmoc-Gly-OH, fmoc-Ala-OH, fmoc-Ile-OH, fmoc-Leu-OH, fmoc-Trp (Boc) -OH, fmoc-Gln (Trt) -OH, fmoc-Val-OH, fmoc-Phe-OH, fmoc-Lys (COOtBu (CH-Pro-OH) in sequence ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OtBu) -Ala-OH, fmoc-Gln (Trt) -OH, fmoc-Ala-OH, fmoc-Ile-OH, fmoc-Lys (Boc) -OH, fmoc-Asp (OtBu) -OH, fmoc-Leu-OH, fmoc-Aib-OH, fmoc-Ile-OH, fmoc-Ser (tBu) -OH, fmoc-Tyr (tBu) -OH, fmoc-Asp (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Thr (tBu) -OH, fmoc-Phe-Thr (tBu) -OH, fmoc-Gly-OH, fmoc-Glu (OtBu) -OH, fmoc-Aib-OH and Boc-Tyr (tBu) -OH. The peptide resin synthesis was completed, with 4 washes with DMF, 3 washes with DCM, 3 washes with MeOH, and drying to give Tirzepatide resin a: boc-Tyr (tBu) -Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Tyr (tBu) -Ser (tBu) -Ile-Aib-Leu-Asp (OtBu) -Lys (Boc) -Ile-Ala-Gln (Trt) -Lys (COOtBu (CH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OtBu) -Ala-Phe-Val-Gln (Trt) -Trp (Boc) -Leu-Ile-Ala-Gly-Gly-Pro-Ser (tBu) -Ser (tBu) -Gly-Ala-Pro-Pro-Ser (tBu) -Rink Amide MBHA resin, the weight was 208.41g.

(2) Cleavage of the Tirzepatide peptide resin

208.41g of the Tirzepatide peptide resin A are weighed and 2084.0ml of cleavage reagent TFA/Tis/H is added ₂ O/Mpr/m-Cresol =94/1/1/2/2, and the cleavage reaction is carried out for 1-4 h at 25 +/-5 ℃. After the reaction was complete, filtration was performed and the resin was washed with 200ml TFA. Adding the filtrate into 20L of frozen isopropyl ether for sedimentation, separating out solids, centrifuging, and washing to obtain a Tirzepatide crude product: H-Tyr-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Aib-Leu-Asp-Lys-Ile-Ala-Gln-Lys (COOtBu (CH) ₂ ) ₁₈ CO-γ-Glu-AEEA-AEEA)-Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH ₂ The weight is 96.08g, the yield is 96.75, the purity is 78.9 percent, and the maximum single impurity is less than 5.0 percent.

(3) Purification of the crude Tirzepatide

Dissolving the Tirzepatide crude product in 10% acetonitrile water solution, filtering with 0.45 μm organic membrane, purifying and separating with reversed phase liquid chromatography, wherein the filler is reversed phase C18, particle size is 8 μm, and pore size is

Gradient elution was performed using 0.1% aqueous TFA and acetonitrile to give the Tirzepatide protide fine peptide in a weight of 28.02g, yield of 32.6%, purity of 99%, and maximum single impurity of less than 0.2%.

Example 14

Tirzepatide synthesis:

(1) Synthesis of Tirzepatide peptide resin B

Add 20% of PIPE/DMF to remove Fmoc protection twice, deprotection times were 5min +15min, respectively, DMF washes 6 times. Fmoc-Ser (tBu) -OH (30.68g, 4.0eq.) and HOBt (11.89g, 4.4q.) are weighed and dissolved in 100ml of DMF, DIC (15.15ml, 6.0eq.) is added at the temperature of 0-10 ℃ for activation for 4min, the mixture is added into a synthesis reaction column, and the reaction is carried out for 2-4h under the protection of nitrogen at the temperature of 25 +/-5 ℃. It was drained and washed 4 times with DMF.

Repeating the above steps, coupling Fmoc-Pro-OH, fmoc-Ala-OH, fmoc-Gly-OH, fmoc-Ser (tBu) -OH, fmoc-Pro-OH, fmoc-Gly-OH, fmoc-Ala-OH, fmoc-Ile-OH, fmoc-Leu-OH, fmoc-Trp (Boc) -OH, fmoc-Gln (Trt) -OH, fmoc-Val-OH, fmoc-Phe-OH, alloc-Lys (COOtBu (CH-Pro-OH), fmoc-Pro-OH, fmoc-Ala-Gly-OH, fmoc-Ile-OH, fmoc-Leu-OH, fmoc-Trp (Boc) -OH, fmoc-Gln (Trt) -OH, fmoc-Val-OH, and Alloc-Lys (COOtBu (CH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OMe) -Ala-OH, coupling was completed 4 times with DMF washes;

alloc protection was removed twice by addition of phenylsilane (21.64g, 10.0 q.) and tetrakis (triphenylphosphine) palladium (2.31g, 0.1q.) and deprotection was performed30min, and DMF washing for 6 times. Repeating the amino acid coupling and the removal of Fmoc protecting group, and sequentially continuing to couple Fmoc-Gln (Trt) -OH, fmoc-Ala-OH, fmoc-Ile-OH, fmoc-Lys (Boc) -OH, fmoc-Asp (OtBu) -OH, fmoc-Leu-OH, fmoc-Aib-OH, fmoc-Ile-OH, fmoc-Ser (tBu) -OH, fmoc-Tyr (tBu) -OH, fmoc-Asp (OtBu) -OH, fmoc-Ser (tBu) -OH, fmoc-Thr (tBu) -OH, fmoc-Gly-OH, fmoc-Glu (OtBu) -OH, fmoc-Aib-OH and Boc-Tyr (tBu) -OH. The peptide resin synthesis was completed, 4 times with DMF, 3 times with DCM, 3 times with MeOH, and dried to give Tirzepatide resin a: boc-Tyr (tBu) -Aib-Glu (OtBu) -Gly-Thr (tBu) -Phe-Thr (tBu) -Ser (tBu) -Asp (OtBu) -Tyr (tBu) -Ser (tBu) -Ile-Aib-Leu-Asp (OtBu) -Lys (Boc) -Ile-Ala-Gln (Trt) -Lys (COOtBu (CH) ₂ ) ₁₈ CO-Glu (AEEA-AEEA) -OMe) -Ala-Phe-Val-Gln (Trt) -Trp (Boc) -Leu-Ile-Ala-Gly-Gly-Pro-Ser (tBu) -Ser (tBu) -Gly-Ala-Pro-Pro-Ser (tBu) -Rink Amide MBHA resin, the weight was 206.85g.

(2) Cleavage of the Tirzepatide peptide resin

206.85g of the Tirzepatide peptide resin B was weighed and 2068.5ml of cleavage reagent TFA/Tis/H was added ₂ O/Mpr/m-Cresol =94/1/1/2/2, and the cleavage reaction is carried out for 1-4 h at 25 +/-5 ℃. After the reaction was complete, filtration was carried out and the resin was washed with 200ml TFA. Adding the filtrate into 20L of frozen isopropyl ether for sedimentation, separating out solid, centrifuging and washing to obtain a crude product of the Tirzepatide with Lys side chain gamma-Glu with OMe protecting group: H-Tyr-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ile-Aib-Leu-Asp-Lys-Ile-Ala-Gln-Lys (COOtBu (CH) ₂ ) ₁₈ CO-Glu(AEEA-AEEA)-OMe)-Ala-Phe-Val-Gln-Trp-Leu-Ile-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH ₂ The weight was 97.43g.

(3) Tirzepatide methyl removing esterification and purification refining

97.43g of crude Tirzepatide with OMe protecting group Lys side chain gamma-Glu was weighed and added to 1000ml acetonitrile: h ₂ A mixed solution of O = 1. Slowly adding LiOH until the pH value of the reaction solution is 9.0-11.0, and stirring to react for 8-16h. After the reaction is finished, the pH value of the reaction solution is adjusted to 6.0-7.0 by dilute hydrochloric acid, and the reaction solution is purified. The aqueous acetonitrile solution of the crude Tirzepatide was diluted with 5L of water and applied to a 0.45 μm organic filmFiltering, purifying and separating with reversed phase liquid chromatography, wherein the filler is reversed phase C18, particle diameter is 8 μm, and pore diameter is

Gradient elution was performed using 0.1% aqueous TFA and acetonitrile to give the Tirzepatide refined peptide in a weight of 27.86g, yield of 30.2%, purity of 99%, and maximum single impurity of less than 0.2%.

Claims

1. A peptide compound characterized by: has a structure shown in a general formula I, and the structure is as follows:

wherein the content of the first and second substances,

R3 is selected from hydrogen, ester protecting group,

m is 10 to 20 ₅ Is COOH or carboxyl with a protecting group,

n is 1 or 2, and n is a hydrogen atom,

or enantiomers, diastereomers and salts thereof.

2. The peptide compound of claim 1, wherein R1 is selected from the group consisting of H, fmoc, dde, alloc, boc, moz, trt, dmb, mmt, mtt.

3. The peptide compound of claim 1, wherein R1 is selected from COCH (CH 3) NH ₂ 、COCH(CH3)NH ₂ -Fmoc、COCH(CH3)NH ₂ -Dde、COCH(CH3)NH ₂ -Alloc、COCH(CH3)NH ₂ -Boc、COCH(CH3)NH ₂ -Moz、COCH(CH3)NH ₂ -Trt、COCH(CH3)NH ₂ -Dmb、COCH(CH3)NH ₂ -Mmt、COCH(CH3)NH ₂ -Mtt。

4. The peptide compound of claim 1, wherein R2 is selected from the group consisting of OH, otBu, OCH ₃ 、OCH2CH ₃ 、OBzl、OCH ₂ CH＝CH ₂ 、OCH ₂ C ₆ H ₅ 。

5. The peptidal compound of claim 1, wherein R2 is selected from NHCH (CH) ₃ )COOH、NHCH(CH ₃ )COOtBu、NHCH(CH ₃ )COOCH3、NHCH(CH ₃ )COOCH ₂ CH ₃ 、NHCH(CH ₃ )COOBzl、NHCH(CH ₃ )COOCH ₂ CH＝CH ₂ 、NHCH(CH ₃ )COOCH ₂ C ₆ H ₅ 。

6. The peptide compound of claim 1, wherein R3 is selected from the group consisting of H, tBu, and CH ₃ 、CH ₂ CH ₃ 、Bzl、CH ₂ CH＝CH ₂ 、CH ₂ C ₆ H ₅ 。

7. The peptide compound of claim 1, wherein R4 is selected from H, fmoc, dde, alloc, boc, moz, trt, dmb, mmt, or Mtt.

8. The peptide compound of claim 1, wherein m is 14 to 20.

9. The peptide compound of claim 1, wherein m is 16.

10. The peptide compound of claim 1, wherein m is 18.

11. The peptide compound of claim 1, wherein R5 is selected from the group consisting of COOH, COOtBu, COOCH ₃ 、COOCH ₂ CH ₃ 、COOBzl、COOCH ₂ CH＝CH ₂ 、COOCH ₂ C ₆ H ₅ 。

12. The peptide compound of claim 1, wherein R4 is selected from CO (CH) ₂ ) ₁₆ COOH、CO(CH ₂ ) ₁₈ COOH、CO(CH ₂ ) ₁₆ COOtBu、CO(CH ₂ ) ₁₈ COOtBu。

13. The peptide compound of claim 1, n is 2.

14. The peptide compound of claim 1, wherein R1 is Fmoc and R2 is NHCH (CH) ₃ ) COOH, R3 is tBu, R4 is CO (CH) ₂ ) ₁₈ COOtBu, n is 2.

15. The peptidal compound of claim 1, wherein R1 is Alloc and R2 is NHCH (CH) ₃ ) COOH, R3 is Me, R4 is CO (CH) ₂ ) ₁₈ COOtBu, n is 2.

16. The peptidal compound of claim 1, wherein R1 is NHCH (CH) ₃ ) CO-Fmoc, R2 is OH, R3 is tBu, R4 is CO (CH) ₂ ) ₁₆ COOtBu, n is 2.

17. The peptidal compound of claim 1, wherein R1 is NHCH (CH) ₃ ) CO-Dde, R2 is OH, R3 is tBu, R4 is CO (CH) ₂ ) ₁₆ COOBzl, n is 2.

18. Synthesis of a peptide compound according to claim 1 comprising linking to the Lys-moiety of the peptide chain (AEEA) _n -gamma Glu-a peptide of fatty alkanedioic acid side chain building blocks.

19. The use of claim 18, wherein the synthesis is solid phase peptide synthesis.

20. Use of a peptidic compound according to claim 1 for the preparation of a medicament of an antidiabetic polypeptide.

21. The use of a peptide compound of claim 20 for the preparation of an anti-diabetic polypeptide medicament, wherein: the antidiabetic polypeptide medicament is a GLP-1 analogue or a GIP/GLP-1 analogue.

22. The GLP-1 analog of claim 21 which is Semaglutide.

23. The GIP/GLP-1 analog of claim 21 which is Tirzepatide.