CN110642923B - Cyclic peptide with antibacterial activity and synthetic method thereof - Google Patents

Cyclic peptide with antibacterial activity and synthetic method thereof Download PDF

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CN110642923B
CN110642923B CN201911005981.8A CN201911005981A CN110642923B CN 110642923 B CN110642923 B CN 110642923B CN 201911005981 A CN201911005981 A CN 201911005981A CN 110642923 B CN110642923 B CN 110642923B
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cys
cyclic peptide
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antibacterial activity
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权春善
张丽影
吴亚多
白鹏阳
陈苛蒙
范圣第
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Dalian Minzu University
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Abstract

The invention relates to a cyclic peptide with antibacterial activity and a synthesis method thereof, belonging to the technical field of biological pharmacy. The method adopts the main technical scheme that under the room temperature and slightly alkaline environment, m-dibromide benzyl is used as a micromolecular auxiliary reagent, and the thiol of two cysteine in the linear peptide is subjected to nucleophilic substitution reaction with the thiol, so that the linear peptide is cyclized. The method for cyclizing the linear chain peptide has mild reaction conditions and is easy to realize. The reaction speed is high, the time is short, and the reaction can be finished within 2 hours. The cyclization efficiency can reach more than 90 percent, the total yield of the cyclic peptide can reach more than 60 percent, and the synthesis efficiency of the cyclic peptide is greatly improved. The prepared cyclic peptide has high antibacterial activity.

Description

Cyclic peptide with antibacterial activity and synthetic method thereof
Technical Field
The invention belongs to the field of biological pharmacy, and particularly relates to a cyclic peptide with antibacterial activity and a synthesis method thereof.
Background
The problem of bacterial resistance has been increasing with the long-term use or abuse of antibiotics, particularly broad-spectrum antibiotics, since the 20 th century. Bacterial resistance has become a global concern, and the evolution and infection of resistant bacteria poses new challenges for the prevention and treatment of bacterial infectious diseases, which will become a major disease threatening human health. The american society for infectious diseases research reports that drug-resistant bacterial infection is associated with a significant increase in mortality, and statistically, drug-resistant infection causes 200 million patients per year in the united states, with about 23000 dying. Therefore, there is an urgent need to develop new, highly effective, safe antibacterial agents to cope with serious pathogenic infection.
The cyclic lipopeptide compound has wide application prospect in the field of medicine due to the unique structural characteristics and biological activity. The cyclic lipopeptide compound has broad-spectrum antibacterial activity, the antibacterial mechanism of the cyclic lipopeptide compound is different from that of any type of antibiotics on the market at present, and the cyclic lipopeptide compound has the characteristics of high efficiency, low toxicity and difficulty in drug resistance generation clinically.
Bacillus D (spore peptide) is a main member of Iturin family cyclic lipopeptide, and is separated and extracted from Bacillus subtilis fermentation liquor for the first time in 1980 and the chemical structure of the Bacillus subtilis fermentation liquor is determined. Bacillus D has strong inhibiting effect on yeast and filamentous fungi and good anti-tumor activity. Bacillus lacticola D is formed by cyclizing beta-amino fatty acid consisting of 14-17 carbon atoms and a peptide chain consisting of 7 amino acids through amido bonds, the molecular weight is about 1000, and the amino acid sequence in the peptide chain is as follows: L-Asn-D-Tyr-D-Asn-L-Gln-L-Pro-D-Asn-L-Ser, the structure diagram is shown in figure 18.
Until now, bacillus subtilis D is obtained by microbial fermentation, but the yield is low, and various cyclic lipopeptides or homologues of the same cyclic lipopeptides exist in metabolites, so that the microbial fermentation method for obtaining a large amount of high-purity bacillus subtilis D meets the bottleneck, and the research and application of the bacillus subtilis D in the field of medicine are hindered. By adopting a chemical synthesis method, the natural cyclic lipopeptide is subjected to structural modification to hopefully obtain the cyclic peptide drug which has high biological activity and can be industrially produced.
The synthesis of linear peptides is well established, but the synthesis of cyclic peptides is much more difficult. The conventional methods for cyclizing a linear peptide mainly include an active ester method, an azide method, a mixed acid anhydride method, a thioester method, and the like, but these methods have certain disadvantages. Such as: the active ester method is to prepare active ester from carboxyl, remove a nitrogen end protecting group and cyclize, but the active ester has large stereoscopic effect and low reactivity in the reaction process, so the cyclization speed is slow. The azide method is characterized in that carboxyl terminals are prepared into acyl azide active intermediates, and then cyclization is carried out in a dilute alkaline solution, wherein the azide method has higher efficiency than an active ester method, but the acyl azide intermediates are unstable, the azido acid is toxic, and the preparation steps are very complicated. The mixed anhydride method is to prepare mixed anhydride from carboxyl terminal and then cyclize in diluted alkaline solution, because the reactive intermediate formed in the reaction is extremely unstable, the temperature is strictly kept low during the preparation of anhydride, and the used instruments and solvents need to be absolutely dried. The thioester method is to utilize the nucleophilicity of sulfhydryl to attack carbonyl on thioester bond at carbon end by cysteine sulfhydryl at nitrogen end to form cyclic thioester in molecule, and then form cyclic peptide connected by amide bond through bond type conversion from S to N.
Disclosure of Invention
The invention provides a cyclic peptide with antibacterial activity and a synthesis method thereof. The linear peptide cyclization reaction condition is mild, the reaction time is short, the cyclization efficiency is high, and the synthesis efficiency of cyclic peptide can be greatly improved. The technical scheme of the invention is as follows:
a cyclic peptide with antibacterial activity is prepared from the thiol groups of two cysteines of straight-chain peptide and m-dibromobenzyl through nucleophilic substitution reaction and cyclizing.
Further, the linear peptide has the structural formula:
Figure BDA0002242787420000021
the structural formula of the cyclic peptide is:
Figure BDA0002242787420000022
further, the linear peptide has the structural formula:
Figure BDA0002242787420000031
the structural formula of the cyclic peptide is:
Figure BDA0002242787420000032
further, the linear peptide has the structural formula:
Figure BDA0002242787420000033
the structural formula of the cyclic peptide is:
Figure BDA0002242787420000034
further, the linear peptide has the structural formula:
Figure BDA0002242787420000041
the structural formula of the cyclic peptide is:
Figure BDA0002242787420000042
in the above four structural formulas, the groups represented by R are the same, and are specifically as follows:
position 1 (R) 1 ): DL-alanine; DL-arginine; DL-aspartic acid; DL-glutamine; DL-glutamic acid; DL-histidine; DL-isoleucine; DL-glycine; DL-asparagine; DL-leucine; DL-lysine; DL-methionine; DL-phenylalanine; DL-proline; DL-serine; DL-threonine; DL-tryptophan; DL-tyrosine; DL-valine;
position 2 (R) 2 ): DL-alanine; DL-arginine; DL-aspartic acid; DL-glutamine; DL-glutamic acid; DL-histidine; DL-isoleucine; DL-glycine; DL-asparagine; DL-leucine; DL-lysine; DL-methionine; DL-phenylalanine; DL-proline; DL-serine; DL-threonine; DL-tryptophan; DL-tyrosine; DL-valine;
position 3 (R) 3 ): DL-alanine; DL-arginine; DL-aspartic acid; DL-glutamine; DL-glutamic acid; DL-histidine; DL-isoleucine; DL-glycine; DL-asparagine; DL-leucine; DL-lysine; DL-methionine; DL-phenylalanine; DL-proline; DL-serine; DL-threonine; DL-tryptophan; DL-tyrosine; DL-valine;
position 4 (R) 4 ): DL-alanine; DL-arginine; DL-aspartic acid; DL-glutamine; DL-glutamic acid; DL-histidine; DL-isoleucine; DL-glycine; DL-asparagine; DL-leucine;DL-lysine; DL-methionine; DL-phenylalanine; DL-proline; DL-serine; DL-threonine; DL-tryptophan; DL-tyrosine; DL-valine;
position 5 (R) 5 ): DL-alanine; DL-arginine; DL-aspartic acid; DL-glutamine; DL-glutamic acid; DL-histidine; DL-isoleucine; DL-glycine; DL-asparagine; DL-leucine; DL-lysine; DL-methionine; DL-phenylalanine; DL-proline; DL-serine; DL-threonine; DL-tryptophan; DL-tyrosine; DL-valine;
position 6 (R) 6 ): DL-alanine; DL-arginine; DL-aspartic acid; DL-glutamine; DL-glutamic acid; DL-histidine; DL-isoleucine; DL-glycine; DL-asparagine; DL-leucine; DL-lysine; DL-methionine; DL-phenylalanine; DL-proline; DL-serine; DL-threonine; DL-tryptophan; DL-tyrosine; DL-valine;
position 7 (R) 7 ): DL-alanine; DL-arginine; DL-aspartic acid; DL-glutamine; DL-glutamic acid; DL-histidine; DL-isoleucine; DL-glycine; DL-asparagine; DL-leucine; DL-lysine; DL-methionine; DL-phenylalanine; DL-proline; DL-serine; DL-threonine; DL-tryptophan; DL-tyrosine; DL-valine;
position 8 (R) 8 )：CH 3 (CH 2 ) n COOH, n is any integer from 1 to 18; CH (CH) 3 (CH 2 ) n COOCl, n is any integer from 1 to 18; sorbic acid, trans-10-hydroxy-2-decenoic acid, trans-2-hexenoic acid, dodecenoic acid, decatetraenoic acid, oleic acid, linoleic acid, elaidic acid, arachidonic acid, cis-9-hexadecenoic acid, 11-dodecenoic acid, 10-undecenoic acid.
Further, the cyclic peptide adjusts the structure of the cyclic peptide by changing the position of cysteine.
The invention simultaneously discloses a preparation method of the cyclic peptide with antibacterial activity, which comprises the steps of preparing the linear peptide by utilizing a Fomc solid phase synthesis method, and then carrying out nucleophilic substitution reaction on sulfydryl of two cysteines in the linear peptide and m-dibromide benzyl to cyclize the linear peptide.
The invention has the following beneficial effects:
(1) in the invention, m-dibromide benzyl is used as a micromolecular auxiliary reagent under room temperature and slightly alkaline environment, and the thiol groups of two cysteine in the linear peptide are subjected to nucleophilic substitution reaction with the thiol groups, so that the linear peptide is cyclized. The method for cyclizing the linear chain peptide has mild reaction conditions and is easy to realize. The reaction speed is high, the time is short, and the reaction can be finished within 2 hours. The cyclization efficiency can reach more than 90 percent, the total yield of the cyclic peptide can reach more than 60 percent, and the synthesis efficiency of the cyclic peptide is greatly improved.
(2) The prepared cyclic peptide has high antibacterial activity.
Drawings
FIG. 1 Linear CC 19 High performance liquid chromatogram map
(Linear CC) 19 Peptide sequence: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Ser (D) -Thr-Cys);
FIG. 2 Linear chain CC 19 Mass spectrogram;
FIG. 3Cyclo-CC 19 High performance liquid chromatogram;
FIG. 4Cyclo-CC 19 Mass spectrogram;
FIG. 5 Linear chain CC 18 High performance liquid chromatogram map
(Linear CC) 18 Peptide sequence: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Ser (D) -Cys-Thr);
FIG. 6 Linear CC 18 Mass spectrogram;
FIG. 7Cyclo-CC 18 High performance liquid chromatogram;
FIG. 8Cyclo-CC 18 Mass spectrogram;
FIG. 9 straight-chain CC 17 High performance liquid chromatogram;
(Linear CC) 17 Peptide sequence: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Cys-Ser (D) -Thr);
FIG. 10 Linear chain CC 17 Mass spectrogram;
FIG. 11Cyclo-CC 17 High performance liquid chromatogram;
FIG. 12Cyclo-CC 17 Mass spectrogram;
FIG. 13 straight-chain CC17 high performance liquid chromatogram;
(linear CC16 peptide sequence Cys-Asp-Tyr (D) -Gln (D) -Pro-Cys-Glu-Ser (D) -Thr);
FIG. 14 Linear CC 17 Mass spectrogram;
FIG. 15Cyclo-CC 17 High performance liquid chromatogram map;
FIG. 16Cyclo-CC 17 Mass spectrogram;
FIG. 17 shows the bacteriostatic effect of cyclic peptide;
FIG. 18 is a schematic view of Bacillus subtilis D (bacitracin).
Detailed Description
The invention will be further illustrated and explained with reference to specific examples.
Example 1: Cyclo-CC 19 Synthesis of (2)
Figure BDA0002242787420000071
The linear peptide amino acid sequence was changed to: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Ser (D) -Thr-Cys
Cyclized Cyclo-CC 19 The structure is as follows:
Figure BDA0002242787420000072
(1) linkage of the first amino acid
1g of dried 2-CTC resin was placed in a reaction kettle and 15ml of DCM was added and swollen for 60 min. After removal of DCM, swelling was continued with 15ml of DMF for 60 min. After DMF was removed, the resin was placed in 10ml DCM, and at a ratio of Fomc-amino acid/resin of 3mmol/g and a ratio of condensing agent DIEA/resin of 17mmol/g, Fomc-cys (trt) -OH 1.758g and DIEA 2.805ml were added, respectively, stirred at room temperature for 2h, followed by addition of DIEA/methanol (1: 9) solution to block the unreacted site for 20 min. After blocking, the resin was filtered and washed with DCM, DMF, DCM and MeOH in that order and dried in vacuo.
(2) Detection of degree of amino acid substitution
The resin with the amino acid attached is weighed to be m 0.0157g, and is dissolved in 4ml of 25% piperidine/DMF solution, and blank control with the same proportion is prepared at the same time. The sample group and the control group were simultaneously diluted 100-fold, and the absorbance at 300.5nm of the samples in the two groups was measured using a quartz cuvette, and the amino acid substitution degree was (0.1293 × 0.138-0.0014) × 0.4/0.0157 was 0.42 mmol/g.
(3) Straight chain CC 19 Synthesis of (2)
0.8g of 2-CTC-Cys-Fmoc resin was weighed into a reaction vessel, swollen with 15ml of DCM for 60min, filtered off the DCM and swollen with 15ml of DMF for 60 min. After swelling, detection using (ninhydrin + n-butanol + acetic acid) solution showed no removal of Fmoc protecting group as colorless. And (3) adding 15ml of 20% piperidine/DMF (dimethyl formamide) to remove the Fmoc protecting group of the amino acid, reacting for 20min, and detecting by using a (ninhydrin + n-butanol + acetic acid) solution after the reaction is finished, wherein the condition is bluish purple, which indicates that the Fmoc protecting group is completely removed. Washing with 15ml DMF, 15ml DCM, 15ml methanol, 15ml DMF twice in sequence, transferring the resin into 15ml DMF, adding the next amino acid and the condensing agents HBTU and HOBT in proportion, and adding DIEA to start the reaction after 10 min. Reaction time: 40min, reaction temperature: at 35 deg.c. And repeating the steps in sequence until all the amino acids are connected. The peptide chain-linked resin was dried in vacuo and weighed 0.84 g/m.
TABLE 1 amount of reactants added
Figure BDA0002242787420000081
Figure BDA0002242787420000091
(4) Straight chain CC 19 Detection of degree of substitution
Weighing connected straight-chain CC 19 The resin mass of (1) was 0.0088g, dissolved in 4ml of 25% piperidine/DMF solution, while preparing a blank in the same ratio. The sample group and the control group were simultaneously diluted 100-fold, and the absorbance at 300.5nm of the samples in the two groups was measured using a quartz cuvette, and the amino acid substitution degree was (0.1293 × 0.138-0.0014) × 0.4/0.0157 was 0.42 mmol/g.The degree of amino acid substitution was (0.1293 × 0.079-0.0014) × 0.4/0.0088 was 0.4 mmol/g.
(5) Straight chain CC 19 Cutting of (2)
0.84g of the peptide chain-linked resin was placed in a round-bottomed flask, 20ml of a solution precooled at-20 ℃ (TFA: m-cresol: ethanedithiol 95%: 1%: 4%) was added, and the mixture was reacted in an ice-water bath for 30min and then at room temperature for 3 hours. After the reaction was complete, the resin was filtered and 50ml of glacial ethyl ether was added to the filtrate and precipitated overnight at-20 ℃. The mixture was centrifuged at 6000rpm at 4 ℃ for 5min, the precipitate was collected and dissolved in 1ml of 50% acetonitrile/water, filtered through a 0.22um organic membrane, and subjected to HPLC.
(6) Straight chain CC 19 LC-MS detection
LC-MS detection of Linear CC 19 Column model Agela Venusil ASB C18, size 4.6 x 250mm, mobile phase a water containing 0.1% (vol/vol) trifluoroacetic acid; mobile phase B acetonitrile containing 0.1% (vol/vol) trifluoroacetic acid. The flow rate is 0.3mL/min, the detection wavelength is 220nm, and the B phase is reduced from 90% to 59.5% in 30 min. Identification of the structure by LC-MS (see FIGS. 1 and 2)
(7) Straight chain CC 19 Liquid phase preparation
Purification was performed by means of waters preparative liquid chromatography. Column model Agilent ZORBAX SB-C18, size 9.4 x 250mm, mobile phase A: water containing 0.1% (volume concentration) trifluoroacetic acid; mobile phase B: acetonitrile containing 0.1% (vol/vol) trifluoroacetic acid. Keeping 90% of phase B and 10% of phase A, eluting for 10min at equal gradient with flow rate of 5mL/min and detection wavelength of 220 nm. The prepared linear peptide was lyophilized to give the final product in a purity of 95% or more (95.2% in this example).
(8) Straight chain CC 19 By cyclization of
18mg of the linear peptide was dissolved in 1ml of a 50% acetonitrile/water solution, followed by dissolving 2.56mg of ammonium carbonate in 100. mu.l of water having a pH of 8.6, 200. mu.M of tris (2-carboxyethyl) phosphine hydrochloride (TCEP) in water, 3.86mg of M-dibromide in 200. mu.l of acetonitrile, mixing the above solutions, adjusting the system pH to 8.6, and reacting with stirring at room temperature for 120 min. After the reaction was completed, the mixture was dialyzed 3 times against water having a pH of 8.6. Filtering the dialysate with 0.22um organic membrane, and freeze drying.
(9)Cyclo-CC 19 LC-MS detection
A small amount of cyclopeptide powder was dissolved in 50% acetonitrile/water and detected by LC-MS. Column model Agela Venusil ASB C18, size 4.6 x 250mm, mobile phase a water containing 0.1% (vol/vol) trifluoroacetic acid; mobile phase B acetonitrile containing 0.1% (vol/vol) trifluoroacetic acid. The flow rate is 0.3mL/min, the detection wavelength is 220nm, and the B phase is reduced from 90% to 59.5% in 30min, and the structure is identified by LC-MS (shown in figure 3 and figure 4).
(10) Preparation of cyclic peptides
Purifying by using waters preparative liquid chromatography. Column model Agilent ZORBAX SB-C18, size 9.4 x 250mm, mobile phase A: water containing 0.1% (volume concentration) trifluoroacetic acid; mobile phase B: acetonitrile containing 0.1% (vol/vol) trifluoroacetic acid. Keeping 90% of phase B and 10% of phase A, eluting for 10min at equal gradient with flow rate of 5mL/min and detection wavelength of 220 nm. And (3) freeze-drying the prepared sample to obtain the cyclic peptide. The purity was more than 95% (95% in this example).
Example 2: Cyclo-CC 18 Synthesis of (2)
The synthetic steps are the same as those of Cyclo-CC 19 Only the linear peptide amino acid sequence was changed to:
Cys-Asp-Tyr(D)-Gln(D)-Pro-Glu-Ser(D)-Cys-Thr
liquid phase and mass spectra are shown in figures 5 and 6,
cyclized Cyclo-CC 18 The structure is as follows:
Figure BDA0002242787420000111
the liquid phase and mass spectra are shown in FIGS. 7 and 8.
Example 3: Cyclo-CC 17 Synthesis of (2)
The synthetic steps are the same as those of Cyclo-CC 19 Only the linear peptide amino acid sequence was changed to:
Cys-Asp-Tyr(D)-Gln(D)-Pro-Glu-Cys-Ser(D)-Thr
the liquid phase and mass spectra are shown in FIGS. 9 and 10.
Cyclized Cyclo-CC 17 The structure is as follows:
Figure BDA0002242787420000112
the liquid phase and mass spectra are shown in FIGS. 11 and 12.
Example 4: Cyclo-CC 16 Synthesis of (2)
The synthetic steps are the same as those of Cyclo-CC 19 Only the linear peptide amino acid sequence was changed to:
Cys-Asp-Tyr(D)-Gln(D)-Pro-Cys-Glu-Ser(D)-Thr
the liquid phase and mass spectra are shown in FIGS. 13 and 14.
Cyclized Cyclo-CC 16 The structure is as follows:
Figure BDA0002242787420000121
the liquid phase and mass spectra are shown in FIGS. 15 and 16.
EXAMPLE 5 bacteriostatic Activity of Cyclic peptides
(1) Preparation of a culture medium: PDA culture medium formula: 200 g of potato, 20 g of glucose, 15 g of agar, 1000 ml of distilled water, natural pH and sterilization for later use.
(2) Activating strains: the preserved Candida albicans strain was inoculated into a sterilized liquid medium in a super clean bench at an inoculum size of 0.1% by pipetting and cultured overnight in a shaker (180r/min) at 30 ℃.
(3) Activity detection
In a clean bench, 1ml of the activated bacterial liquid is diluted by 3 times by using a PDA culture medium, the OD value is measured, and when the value is 0.5, the bacterial liquid is diluted by 1000 times by using the PDA culture medium for standby.
The cyclopeptide and pure DMSO solvent are diluted to a certain concentration by using PDA culture medium, then the sample is added into a 96-well plate, and 100ul of PDA culture medium is added into each well in the outermost circle, so that the edge effect is prevented. The wells with 90ul of diluted bacterial solution and 10ul of diluted cyclic peptide were used as experimental groups, the wells with 90ul of diluted bacterial solution and 10ul of diluted DMSO were used as control groups, each of the wells was run in parallel three times, and the concentration of DMSO in each well was controlled to be less than one thousandth. After sample application, the cover is closed, the culture is carried out for 10h at 30 ℃ and 180r/min, the OD value of each well is measured, and the cyclic peptide inhibition rate is calculated by the following formula.
Inhibition ratio (OD) Control -OD Experiment of )÷OD Control
As can be seen from FIG. 17, the four cyclic peptides all have obvious bacteriostatic activity, the smaller the peptide loop is, the longer the tail chain is, the stronger the bacteriostatic activity is, wherein 1.25mM of the cyclic peptide Cyclo-CC 16 The activity of inhibiting candida albicans reaches 42.3% in 10 hours.

Claims (7)

1. A cyclic peptide with antibacterial activity is characterized in that the cyclic peptide is formed by cyclization through nucleophilic substitution reaction of sulfydryl of two cysteines of straight-chain peptide and m-dibromobenzyl;
the amino acid sequence of the linear peptide is as follows: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Ser (D) -Thr-Cys, Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Ser (D) -Cys-Thr, Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Cys-Ser (D) -Thr or Cys-Asp-Tyr (D) -Gln (D) -Pro-Cys-Glu-Ser (D) -Thr.
2. A cyclic peptide having antibacterial activity according to claim 1, wherein the amino acid sequence of the linear peptide is: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Ser (D) -Thr-Cys, structural formula of cyclic peptide:
Figure FDA0003752571770000011
3. a cyclic peptide having antibacterial activity according to claim 1, wherein the amino acid sequence of the linear peptide is: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Ser (D) -Cys-Thr, and the structural formula of the cyclic peptide is as follows:
Figure FDA0003752571770000021
4. a cyclic peptide having antibacterial activity according to claim 1, wherein the amino acid sequence of the linear peptide is: Cys-Asp-Tyr (D) -Gln (D) -Pro-Glu-Cys-Ser (D) -Thr, and the structural formula of the cyclic peptide is as follows:
Figure FDA0003752571770000022
5. a cyclic peptide having antibacterial activity according to claim 1, wherein the amino acid sequence of the linear peptide is: Cys-Asp-Tyr (D) -Gln (D) -Pro-Cys-Glu-Ser (D) -Thr, and the structural formula of the cyclic peptide is as follows:
Figure FDA0003752571770000031
6. the cyclic peptide having antibacterial activity according to claim 1, wherein the structure of the cyclic peptide is adjusted by changing the position of cysteine.
7. A method for preparing the cyclic peptide having antibacterial activity according to claim 1, comprising: the preparation method comprises the steps of preparing a linear peptide by using a Fomc solid phase synthesis method, and then cyclizing the linear peptide by using a nucleophilic substitution reaction between sulfydryl of two cysteine in the linear peptide and m-dibrombenzyl.
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