CN108707189A - Antibacterial cyclic peptides and its preparation method and application - Google Patents
Antibacterial cyclic peptides and its preparation method and application Download PDFInfo
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- CN108707189A CN108707189A CN201810499240.9A CN201810499240A CN108707189A CN 108707189 A CN108707189 A CN 108707189A CN 201810499240 A CN201810499240 A CN 201810499240A CN 108707189 A CN108707189 A CN 108707189A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/64—Cyclic peptides containing only normal peptide links
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The invention discloses a kind of antibacterial cyclic peptides and its preparation method and application, the structure of antibacterial cyclic peptides antibacterial cyclic peptides as shown in any one of formula (I-1), formula (I-2), formula (I-3), formula (I-4), formula (I-5), formula (I-6) have superior antibacterial effect, the preparation method has the advantages that process is simple and yield is high simultaneously, so that the antibacterial cyclic peptides can be with a wide range of applications.
Description
Technical field
The present invention relates to cyclic peptide, and in particular, to a kind of antibacterial cyclic peptides and its preparation method and application.
Background technology
Cyclic peptide Desotamide B ((glycine-tryptophan-leucine-D-Leu-valine-asparagine) peptide) from
It is extracted in the actinomyces Streptomyces scopuliridis SCSIO ZJ46 metabolites of South China Sea deep-sea, by Trp-
Six cyclic peptide (J.Nat.Prod.2014,77 (8), 1937-1941) of Leu-D-Leu-Val-Asn-Gly compositions, color ammonia therein
Acid is the indispensable amino acid of cyclic peptide antibacterial skeleton, and is tested to its antibacterial activity, finds it to resistance to methoxy west
Woods staphylococcus epidermis (MRSE) has good inhibiting effect；Cyclic peptide Desotamide B are to methicillin-resistant epidermis grape ball
Bacterium (MRSE) has good inhibiting effect, reaches 16 μ g/mL to the minimal inhibitory concentration of staphylococcus aureus
(J.Nat.Prod.2014,77(8),1937-1941)。
Currently, ascendant trend is in the research of the functional aspects of cyclic peptide Desotamide B both at home and abroad, but about cyclic peptide
The specific reforming direction of Desotamide B is not able to explore how to improve cyclic peptide also in the primary exploratory stage
The antibacterial effect of Desotamide B.
Invention content
The object of the present invention is to provide a kind of antibacterial cyclic peptides and its preparation method and application, which has better
Antibacterial effect, while the preparation method has the advantages that process is simple and yield is high, so that the antibacterial cyclic peptides can have
Have wide practical use.
To achieve the goals above, the present invention provides a kind of antibacterial cyclic peptides, the structure such as formula (I-1) of the antibacterial cyclic peptides,
Shown in any one of formula (I-2), formula (I-3), formula (I-4), formula (I-5), formula (I-6),
The present invention also provides a kind of preparation methods such as above-mentioned antibacterial cyclic peptides, including：
1) matrix resin is soaked in organic solvent, then removes solvent；Then by matrix resin successively with a variety of ammonia
Base acid carries out multiple step-by-step processing and obtains the second resin；Step-by-step processing is：By resin, amino acid, N, N- diisopropylethylamine has
Solvent carries out haptoreaction, then carries out Seal treatment, washing, removing Fmoc protecting groups, washing；
2) the second resin, cutting liquid are subjected to oscillating reactions, then remove solvent and obtains straight chain peptide chain；
3) in the presence of protecting gas, by straight chain peptide chain, 1- hydroxyl -7- azos benzotriazole, (3H-1,2,3- triazoles
Bing [4,5-b]Pyridine -3- oxygroups) three -1- pyrrolidinyls hexafluorophosphates, N-methylmorpholine, organic solvent carry out ring-closure reaction；
Then solvent is removed, then by post-processing primary cyclic peptide is made；
4) primary cyclic peptide and deprotecting regent are subjected to haptoreaction to remove Boc protecting groups, then by system carry out from
Heart sedimentation, drying are to be made antibacterial cyclic peptides；
Wherein, shown in antibacterial cyclic peptides such as formula (I-1) in the case of, the amino acid of step-by-step processing is independent successively in step 2)
For Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-D-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp (Boc)-OH, Fmoc-
Gly-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-2), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Leu-OH、Fmoc-Leu-OH、Fmoc-Trp(Boc)-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH、Fmoc-
Ala-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-3), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Ala-OH、Fmoc-Leu-OH、Fmoc-Val-OH、Fmoc-Asn(Trt)-OH、Fmoc-Gly-OH、Fmoc-Trp
(Boc)-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-4), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Leu-OH、Fmoc-Val-OH、Fmoc-Asn(Trt)-OH、Fmoc-Gly-OH、Fmoc-Trp(Boc)-OH、Fmoc-
Ala-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-5), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Leu-OH、Fmoc-Leu-OH、Fmoc-Ala-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH、Fmoc-Val-
OH；In the case of shown in antibacterial cyclic peptides such as formula (I-6), the amino acid of step-by-step processing stands alone as Fmoc-D- successively in step 2)
Leu-OH、Fmoc-Leu-OH、Fmoc-Trp(Boc)-OH、Fmoc-Ala-OH、Fmoc-Asn(Trt)-OH、Fmoc-Val-OH。
Invention further provides a kind of such as application of the above-mentioned antibacterial cyclic peptides in antibacterial.
In the above-mentioned technical solutions, as shown in Figure 1, what the present invention was combined by Solid-phase synthesis peptides with alanine scanning
Method, use is small, each amino acid on the amino acid substitution Desotamide B without other functional groups, to eliminate spy
Different site upper amino acid side chain is on the active influence of antibacterial, the results showed that the most special site of cyclic peptide Desotamide B is in amino
Sour Asn (asparagine) position, so that the cyclic peptide has superior antibacterial effect, while the preparation method has work
The advantage that sequence is simple and yield is high, so that the antibacterial cyclic peptides can be with a wide range of applications.
Other features and advantages of the present invention will be described in detail in subsequent specific embodiment part.
Description of the drawings
Attached drawing is to be used to provide further understanding of the present invention, an and part for constitution instruction, with following tool
Body embodiment is used to explain the present invention together, but is not construed as limiting the invention.In the accompanying drawings：
Fig. 1 is the preparation principle figure of Desotamide B-1 provided by the invention；
Fig. 2 is the product MS spectrograms of embodiment 1 provided by the invention；
Fig. 3 is the product of embodiment 1 provided by the invention1H spectrograms；
Fig. 4 is the product MS spectrograms of embodiment 2 provided by the invention；
Fig. 5 is the product of embodiment 2 provided by the invention1H spectrograms；
Fig. 6 is the product MS spectrograms of embodiment 3 provided by the invention；
Fig. 7 is the product of embodiment 3 provided by the invention1H spectrograms；
Fig. 8 is the product MS spectrograms of embodiment 4 provided by the invention；
Fig. 9 is the product of embodiment 4 provided by the invention1H spectrograms；
Figure 10 is the product MS spectrograms of embodiment 5 provided by the invention；
Figure 11 is the product of embodiment 5 provided by the invention1H spectrograms；
Figure 12 is the product MS spectrograms of embodiment 6 provided by the invention；
Figure 13 is the product of embodiment 6 provided by the invention1H spectrograms；
Figure 14 is the bacteriostatic activity test result statistical chart of application examples 1 provided by the invention.
Specific implementation mode
The specific implementation mode of the present invention is described in detail below.It should be understood that described herein specific
Embodiment is merely to illustrate and explain the present invention, and is not intended to restrict the invention.
The endpoint of disclosed range and any value are not limited to the accurate range or value herein, these ranges or
Value should be understood as comprising the value close to these ranges or value.For numberical range, between the endpoint value of each range, respectively
It can be combined with each other between the endpoint value of a range and individual point value, and individually between point value and obtain one or more
New numberical range, these numberical ranges should be considered as specific open herein.
The present invention provides a kind of antibacterial cyclic peptides, structure such as formula (I-1), formula (I-2), formula (I-3), the formulas of the antibacterial cyclic peptides
(I-4), shown in any one of formula (I-5), formula (I-6),
In the present invention, the structure of the antibacterial cyclic peptides can be selected at above-mentioned six kinds, but in order to further increase cyclic peptide
Antibacterial effect, it is preferable that shown in the structure of the antibacterial cyclic peptides such as formula (I-1).
The present invention also provides a kind of preparation methods such as above-mentioned antibacterial cyclic peptides, including：
1) matrix resin is soaked in organic solvent, then removes solvent；Then by matrix resin successively with a variety of ammonia
Base acid carries out multiple step-by-step processing and obtains the second resin；Step-by-step processing is：By resin, amino acid, N, N- diisopropylethylamine has
Solvent carries out haptoreaction, then carries out Seal treatment, washing, removing Fmoc protecting groups, washing；
2) the second resin, cutting liquid are subjected to oscillating reactions, then remove solvent and obtains straight chain peptide chain；
3) in the presence of protecting gas, by straight chain peptide chain, 1- hydroxyl -7- azos benzotriazole, (3H-1,2,3- triazoles
Bing [4,5-b]Pyridine -3- oxygroups) three -1- pyrrolidinyls hexafluorophosphates, N-methylmorpholine, organic solvent carry out ring-closure reaction；
Then solvent is removed, then by post-processing primary cyclic peptide is made；
4) primary cyclic peptide and deprotecting regent are subjected to haptoreaction to remove Boc protecting groups, then by system carry out from
Heart sedimentation, drying are to be made antibacterial cyclic peptides；
Wherein, shown in antibacterial cyclic peptides such as formula (I-1) in the case of, the amino acid of step-by-step processing is independent successively in step 2)
For Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-D-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp (Boc)-OH, Fmoc-
Gly-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-2), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Leu-OH、Fmoc-Leu-OH、Fmoc-Trp(Boc)-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH、Fmoc-
Ala-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-3), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Ala-OH、Fmoc-Leu-OH、Fmoc-Val-OH、Fmoc-Asn(Trt)-OH、Fmoc-Gly-OH、Fmoc-Trp
(Boc)-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-4), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Leu-OH、Fmoc-Val-OH、Fmoc-Asn(Trt)-OH、Fmoc-Gly-OH、Fmoc-Trp(Boc)-OH、Fmoc-
Ala-OH；In the case of shown in antibacterial cyclic peptides such as formula (I-5), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Leu-OH、Fmoc-Leu-OH、Fmoc-Ala-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH、Fmoc-Val-
OH；In the case of shown in antibacterial cyclic peptides such as formula (I-6), the amino acid of step-by-step processing stands alone as Fmoc-D- successively in step 2)
Leu-OH、Fmoc-Leu-OH、Fmoc-Trp(Boc)-OH、Fmoc-Ala-OH、Fmoc-Asn(Trt)-OH、Fmoc-Val-OH。
In the step 1) of above-mentioned preparation method, the dosage of each raw material can select in a wide range in soaking process,
But in order to impregnate effect, it is preferable that in step 1), in soaking process, matrix resin, organic solvent amount ratio be
1g：10-20mL.
In the step 1) of above-mentioned preparation method, soaking conditions can select in a wide range, but in order to impregnate effect
Fruit, it is preferable that immersion meets the following conditions：Soaking temperature is 15-35 DEG C, soaking time 30-50min.
In the step 1) of above-mentioned preparation method, the dosage of each raw material in step-by-step processing can be selected in a wide range
It selects, but for yield, it is preferable that in step 1), on the basis of 1g matrix resins, in every step step-by-step processing, N, N- bis-
The dosage of wopropyl ethyl amine is 0.6-0.8g, the dosage of organic solvent is 10-20mL；
In the case of shown in antibacterial cyclic peptides such as formula (I-1), the dosage of Fmoc-Ala-OH is 290-300mg, Fmoc-
The dosage of Val-OH is 330-1000mg, and the dosage of Fmoc-D-Leu-OH is 330-1000mg, and the dosage of Fmoc-Leu-OH is
The dosage of 330-1000mg, Fmoc-Trp (Boc)-OH is 500-1500mg, and the dosage of Fmoc-Gly-OH is 330-1000mg；
In the case of shown in antibacterial cyclic peptides such as formula (I-2), the dosage of Fmoc-D-Leu-OH is 330-340mg, Fmoc-
The dosage of Leu-OH is 330-1000mg, and the dosage of Fmoc-Trp (Boc)-OH is 500-1500mg, the dosage of Fmoc-Gly-OH
Dosage for 330-1000mg, Fmoc-Asn (Trt)-OH is 600-1800mg, and the dosage of Fmoc-Ala-OH is 300-900mg；
In the case of shown in antibacterial cyclic peptides such as formula (I-3), the dosage of Fmoc-D-Ala-OH is 290-300mg, Fmoc-
The dosage of Leu-OH is 330-1000mg, and the dosage of Fmoc-Val-OH is 330-1000mg, the dosage of Fmoc-Asn (Trt)-OH
Dosage for 600-1800mg, Fmoc-Gly-OH is 330-1000mg, and the dosage of Fmoc-Trp (Boc)-OH is 500-
1500mg；
In the case of shown in antibacterial cyclic peptides such as formula (I-4), the dosage of Fmoc-D-Leu-OH is 330-340mg, Fmoc-
The dosage of Val-OH is 330-1000mg, and the dosage of Fmoc-Asn (Trt)-OH is 600-1800mg, the dosage of Fmoc-Gly-OH
Dosage for 330-1000mg, Fmoc-Trp (Boc)-OH is 500-1500mg, and the dosage of Fmoc-Ala-OH is 300-900mg；
In the case of shown in antibacterial cyclic peptides such as formula (I-5), the dosage of Fmoc-D-Leu-OH is 330-340mg, Fmoc-
The dosage of Leu-OH is 330-1000mg, and the dosage of Fmoc-Ala-OH is 300-900mg, and the dosage of Fmoc-Gly-OH is 330-
The dosage of 1000mg, Fmoc-Asn (Trt)-OH is 600-1800mg, and the dosage of Fmoc-Val-OH is 330-1000mg；
In the case of shown in antibacterial cyclic peptides such as formula (I-6), the dosage of Fmoc-D-Leu-OH is 330-340mg, Fmoc-
The dosage of Leu-OH is 330-1000mg, and the dosage of Fmoc-Trp (Boc)-OH is 500-1500mg, the dosage of Fmoc-Ala-OH
Dosage for 300-900mg, Fmoc-Asn (Trt)-OH is 600-1800mg, and the dosage of Fmoc-Val-OH is 330-1000mg.
In the step 1) of above-mentioned preparation method, the catalytic condition in step-by-step processing can be selected in a wide range
It selects, but for the yield of amino acid connection, it is preferable that in every step step-by-step processing, haptoreaction meets the following conditions：Reaction
Temperature is 15-35 DEG C, reaction time 1-1.5h.
In the step 1) of above-mentioned preparation method, the condition of Seal treatment can select in a wide range, but in order to
The site that reaction is had neither part nor lot on resin is closed in time, it is preferable that Seal treatment meets the following conditions：Treatment temperature is 30-
35 DEG C, processing time 30-50min；
In the step 1) of above-mentioned preparation method, the composition and dosage of the middle closed reagent of Seal treatment can be wide
Selection in range, but in order to close the site for having neither part nor lot in reaction on resin in time, it is preferable that using 1g matrix resins as base
Standard, Seal treatment use the volume ratio of addition 10-20mL for 1：The methanol and dichloromethane mixed solution of 4-5 carries out；
In the step 1) of above-mentioned preparation method, Fmoc protecting groups remove the composition of reagent and dosage can be in wide model
Interior selection is enclosed, but in order to improve the removal effect of Fmoc protecting groups, it is preferable that on the basis of 1g matrix resins, remove Fmoc
Use the volume ratio of addition 10-20mL for 1 in the step of protecting group：The piperidines and N,N-dimethylformamide mixed solution of 3-4
It carries out.
In the step 2) of above-mentioned preparation method, the dosage of cutting liquid can select in a wide range, but in order to carry
The cutting effect of high resin, it is preferable that in step 2), on the basis of the second resin made from 1g matrix resins, cutting liquid
Dosage is 15-20mL.
In the step 2) of above-mentioned preparation method, the condition of oscillating reactions can select in a wide range, but in order to
Improve the cutting efficiency of resin, it is preferable that oscillating reactions meets the following conditions：Reaction temperature is -5~0 DEG C, reaction time 2-
4h。
In the step 2) of above-mentioned preparation method, the component of cutting liquid can select in a wide range, but in order to carry
The cutting efficiency of high resin, it is highly preferred that cutting liquid is 1 by volume ratio：0.8-1.2：Trifluoroethanol, the glacial acetic acid and two of 7-9
Chloromethanes forms.
In the step 3) of above-mentioned preparation method, the dosage of each material can select in a wide range, but in order to carry
The yield of high ring-closure reaction, it is preferable that in step 3), on the basis of the straight chain peptide chain of 400mg, 1- hydroxyl -7- azo benzos
The dosage of triazole is 355-365mg, (3H-1,2,3- San Zuobing [4,5-b]Pyridine -3- oxygroups) three -1- pyrrolidinyl hexafluoros
Phosphatic dosage is 1370-1380mg, and the dosage of N-methylmorpholine is 535-540mg, and the dosage of organic solvent is 400-
500mL。
In the step 3) of above-mentioned preparation method, the condition of ring-closure reaction can select in a wide range, but in order to
Improve the yield of ring-closure reaction, it is preferable that in step 3), ring-closure reaction meets the following conditions：Reaction temperature is 15-35 DEG C,
Reaction time is 12-24h.
In the step 3) of above-mentioned preparation method, the condition of post-processing can select in a wide range, but in order to carry
The yield of high ring-closure reaction, it is highly preferred that post-processing is：Using methanol as eluant, eluent, by sephadex post separation, finally go
Except eluant, eluent.
In the step 3) of above-mentioned preparation method, protect the type of gas that can select in a wide range, but in order to carry
The yield of high ring-closure reaction, it is preferable that protection gas is selected from least one of nitrogen, helium and argon gas；
In the step 3) of above-mentioned preparation method, the addition environment of material can select in a wide range, but in order to
Improve the yield of ring-closure reaction, it is preferable that before ring-closure reaction, carried out in the environment for making an addition to -5~0 DEG C of material.
In the step 4) of above-mentioned preparation method, the dosage of each material can select in a wide range, but in order to carry
High Boc protecting groups removal effect, it is preferable that in step 4), on the basis of primary cyclic peptide made from the straight chain peptide chain of 400mg,
The dosage of deprotecting regent is 5-8mL.
In the step 4) of above-mentioned preparation method, catalytic condition can select in a wide range, but in order to
Improve Boc protecting group removal effects, it is preferable that haptoreaction meets the following conditions：Reaction temperature is -5~0 DEG C, the reaction time
For 1.5-2h.
In the step 4) of above-mentioned preparation method, the component of deprotecting regent can select in a wide range, but be
Raising Boc protecting group removal effects, it is preferable that deprotecting regent is 82-83 by volume ratio：4-6：4-6：4-6：The three of 2-3
Fluoroacetic acid, phenol, water, thioanisole and ethylene dithiol ether composition.
In the above preparation method, the specific type of organic solvent can select in a wide range, but in order into one
Step improves its dispersion effect, it is preferable that organic solvent is selected from dichloromethane, n,N-Dimethylformamide, dimethyl sulfoxide (DMSO) and N-
At least one of methyl pyrrolidone.
In the above preparation method, washing the solvent used and washing times can select in a wide range, still
In order to further increase its dispersion effect, it is preferable that it is dichloromethane and/or N, N- dimethyl formyls to wash the reagent used
Amine, the washing times in each washing operation are 3-5 times.
In the above preparation method, the specific type of matrix resin can select in a wide range, but in order into one
Step improves its dispersion effect, it is preferable that described matrix resin is selected from 2- chlorine trityl chloride resin, Wang resins, MBHA resin
At least one of with rink resins.
Invention further provides a kind of such as application of the above-mentioned antibacterial cyclic peptides in antibacterial.
The present invention will be described in detail by way of examples below.In following embodiment, the english abbreviation of part is explained
It is as follows：Fmoc：Fluorenylmethyloxycarbonyl, Boc：Tertiary butyl oxycarbonyl, DMF：N,N-dimethylformamide, DCM：Dichloromethane,
DIPEA：N, N- diisopropylethylamine, HATU：2- (7- azos benzotriazole)-N, N, N',N'Tetramethylurea hexafluorophosphoric acid
Ester, HOAt：1- hydroxyl -7- azos benzotriazole, NMM：N-methylmorpholine, PyAop：(3H-1,2,3- San Zuobings [4,5-b]
Pyridine -3- oxygroups) three -1- pyrrolidinyls hexafluorophosphates,
Fmoc-Asn(trt)-OH：NαFluorenylmethyloxycarbonyl-NδTrityl-asparagine, Fmoc-Val-OH：N- fluorenes first
Oxygen carbonyl-valine, Fmoc-D-Leu-OH：N- fluorenylmethyloxycarbonyls-D-Leu, Fmoc-Leu-OH：N- fluorenylmethyloxycarbonyls-bright
Propylhomoserin, Fmoc-Trp (Boc)-OH：N-alpha- fluorenylmethyloxycarbonyl-N-in- tert-butoxycarbonyl-l-l-tryptophans, Fmoc-Gly-
OH：N- fluorenylmethyloxycarbonyls-glycine；Fmoc-Asn(Trt)-OH：Fluorenylmethyloxycarbonyl-N- trityls-asparagine,
piperidine：Piperidines, TFE：Trifluoroethanol, Acetic：Acetic acid, TFA：Trifluoroacetic acid, OD：Absorbance.
Preparation example 1
Cutting liquid：By trifluoroethanol, glacial acetic acid, DCM according to 1：1：8 volume ratio is prepared.
Preparation example 2
Deprotecting regent：By trifluoroacetic acid, phenol, ultra-pure water, thioanisole, ethylene dithiol ether according to 82.5：5：5：5：2.5
Volume ratio prepare
Embodiment 1 (as shown in Figure 1)
1) first the DCM of 15mL is used to impregnate 2- chlorine trityl chloride resin 1000mg (degree of substitution 1.03mmol/g) at 30 DEG C
0.5h pumps DCM.
The connection of single amino acids is reacted：Before connecing first amino acid, the Fmoc-Ala-OH amino acid of 295.1mg is weighed,
1h is impregnated with DIPEA the and 10mL DCM of 977 μ L to be activated, then is connect resin and reacted 1h at 35 DEG C and obtain compound 2；Then
With the dichloromethane solution of 20% methanol volume ratios of 15mL, 30min is reacted at 30 DEG C, to having neither part nor lot in the site of reaction on resin
It is closed, after reaction, is washed 3 times with DCM and DMF respectively, pump cleaning solution.15mL 25% is added into reaction tube
The DMF solution of piperidines volume ratio, reacts 30min, and removing Fmoc protecting groups wash 5 with DCM and DMF respectively after reaction
Time, pump cleaning solution.
The step of then above-mentioned connection amino acid removes the Fmoc protecting groups of end and then detects is repeated, is sequentially ingressed into
1000mg Fmoc-Val-OH, 1000mg Fmoc-D-Leu-OH, 1000mg Fmoc-Leu-OH, 1500mg Fmoc-Trp
(Boc)-OH, 1000mg Fmoc-Gly-OH obtain compound 4.
2) compound 4 is transferred in 50mL centrifuge tubes, is added the cutting liquid of 15mL, under conditions of 0 DEG C, oscillating reactions
2.5h, then filtrate is filtered into eggplant type bottle.The DCM in filtrate is removed with Rotary Evaporators, oil pump drains remaining liq, is added
2mL toluene, then drain to get to pale yellow powder shape compound 5.
3) it weighs after HOAT 359.4mg are dissolved in 2mL DMF and is transferred in eggplant type bottle, then to weigh PyAOP 1376.7mg molten
It in the DCM of 200mL, is transferred in eggplant type bottle, adds 590.3 μ L of NMM.Eggplant type bottle is placed in 0 DEG C of ice water, constant pressure is used
Dropping funel will be slowly added drop-wise to dissolved with the 200mL DCM of 400mg compounds 5 in eggplant type bottle, and nitrogen protection is added dropwise
Afterwards, eggplant type bottle is taken out from ice water, 25 DEG C of reactions are overnight (12h).Reaction solution is extracted out with Rotary Evaporators, oil pump is drained extra
DMF.Using methanol as eluant, eluent (GE Healthcare Sephadex LH-20), the solvent portions containing polypeptide, rotation are collected
Compound 6 is obtained after dry.
4) compound 6 is placed in 10mL centrifuge tubes, addition 5mL deprotecting regents, under conditions of 0 DEG C, oscillating reactions
1.5h.Centrifugal sedimentation 3 times, nitrogen drying, obtained white powder compound, with efficient liquid are carried out with the ether of 3 times of volumes
Phase chromatography is further purified, and obtains final product Desotamide B-1 (as shown in formula (I-1)), yield 28%.
Embodiment 2
1) first the DCM of 15mL is used to impregnate 2- chlorine trityl chloride resin 1000mg, (degree of substitution 1.03mmol/ at 30 DEG C
G) 0.5h pumps DCM.
The connection of single amino acids is reacted：Before connecing first amino acid, the Fmoc-D-Leu-OH amino of 335.0mg is weighed
Acid impregnates 1h with DIPEA the and 10mL DCM of 977 μ L and is activated, then meets reaction 1.5h at 30 DEG C of resin and obtain compound
Fmoc-D-Leu-2- chlorine trityl chloride resins.With the dichloromethane solution of 20% methanol volume ratios of 15mL, reacted at 35 DEG C
30min, the site to having neither part nor lot in reaction on resin are closed.After reaction, it is washed 3 times, is pumped with DCM and DMF respectively
Cleaning solution.The DMF solution of 25% piperidines volume ratios of 15mL is added into reaction tube, reacts 30min, removes Fmoc protecting groups.
The step of then above-mentioned connection amino acid removes the Fmoc protecting groups of end and then detects is repeated, is sequentially ingressed into
1000mg Fmoc-Leu-OH, 1500mg Fmoc-Trp (Boc)-OH, 1000mg Fmoc-Gly-OH, 1600mg Fmoc-Asn
(Trt)-OH, 900mg Fmoc-Ala-OH obtain compound Ala-Asn (Trt)-Gly-Trp (Boc)-Leu-D-Leu-2- chlorine
Trityl chloride resin.
2) compound Ala-Asn (Trt)-Gly-Trp (Boc)-Leu-D-Leu-2- chlorine trityl chloride resins are shifted
Into 50mL centrifuge tubes, it is added the cutting liquid of 15mL, under conditions of 0 DEG C, oscillating reactions 2.5h, then filtrate is filtered to eggplant type bottle
In.The DCM in filtrate is removed with Rotary Evaporators, oil pump drains remaining liq, and 2mL toluene is added, then drains to get to yellowish
Color powdered compounds Ala-Asn (Trt)-Gly-Trp (Boc)-Leu-D-Leu.
3) it weighs after HOAT 268.4mg are dissolved in 2mL DMF and is transferred in eggplant type bottle, then to weigh PyAOP 1027.9mg molten
It in the DCM of 200mL, is transferred in eggplant type bottle, adds 440.7 μ L of NMM.Eggplant type bottle is placed in 0 DEG C of ice water, constant pressure is used
Dropping funel will dissolved with 400mg compounds Ala-Asn (Trt)-Gly-Trp (Boc)-Leu-D-Leu 200mL DCM it is slow
It is added drop-wise in eggplant type bottle, nitrogen protection, after being added dropwise, eggplant type bottle is taken out from ice water, 25 DEG C of reactions are overnight (12h).With rotation
Turn evaporimeter to extract reaction solution out, oil pump drains extra DMF.Using methanol as eluant, eluent, with sephadex column (GE
Healthcare Sephadex LH-20) separation, the solvent portions containing polypeptide are collected, compound Huan [ is obtained after being spin-dried for;-
Ala-Asn(Trt)-Gly-Trp(Boc)-Leu-D-Leu-]。
4) by compound Huan [-Ala-Asn(Trt)-Gly-Trp(Boc)-Leu-D-Leu-]It is placed in 10mL centrifuge tubes,
The deprotecting regent of 5mL, under conditions of 0 DEG C, oscillating reactions 2h is added.Centrifugal sedimentation 3 times is carried out with the ether of 3 times of volumes,
Nitrogen dries up, and obtained white powder compound is further purified with high performance liquid chromatography, obtains final product Desotamide
B-2 (as shown in formula (I-2)), yield 46%.
Embodiment 3
1) first the DCM of 15mL is used to impregnate 2- chlorine trityl chloride resin 1000mg, (degree of substitution 1.03mmol/ at 30 DEG C
G) 0.5h pumps DCM.
The connection of single amino acids is reacted：Before connecing first amino acid, the Fmoc-D-Ala-OH amino of 295.1mg is weighed
Acid impregnates 1h with DIPEA the and 10mL DCM of 977 μ L and is activated, then meets reaction 1.5h at 30 DEG C of resin and obtain compound
Fmoc-D-Ala-2- chlorine trityl chloride resins.With the dichloromethane solution of 20% methanol volume ratios of 15mL, 30min is reacted,
It closes in site to having neither part nor lot in reaction on resin.After reaction, it is washed 3 times with DCM and DMF respectively, pumps cleaning solution.
The DMF solution of 25% piperidines volume ratios of 15mL is added into reaction tube, reacts 30min, removes Fmoc protecting groups.
The step of then above-mentioned connection amino acid removes the Fmoc protecting groups of end and then detects is repeated, is sequentially ingressed into
1000mg Fmoc-Leu-OH, 1000mg Fmoc-Val-OH, 1600mg Fmoc-Asn (Trt)-OH, 1000mg Fmoc-
Gly-OH, 1500mg Fmoc-Trp (Boc)-OH obtains compound Trp (Boc)-Gly-Asn (Trt)-Val-Leu-D-Ala-
2- chlorine trityl chloride resins.
2) Trp (Boc)-Gly-Asn (Trt)-Val-Leu-D-Ala-2- chlorine trityl chloride resins are transferred to 50mL
In centrifuge tube, it is added the cutting liquid of 15mL, under conditions of 0 DEG C, oscillating reactions 2.5h, then filtrate is filtered into eggplant type bottle.With
Rotary Evaporators remove the DCM in filtrate, and oil pump drains remaining liq, and 2mL toluene is added, then drain to get to yellowish toner
Last shape compound Trp (Boc)-Gly-Asn (Trt)-Val-Leu-D-Ala.
3) it weighs after HOAT 272.1mg are dissolved in 2mL DMF and is transferred in eggplant type bottle, then to weigh PyAOP 1042.3mg molten
It in the DCM of 200mL, is transferred in eggplant type bottle, adds 446.9 μ L of NMM.Eggplant type bottle is placed in 0 DEG C of ice water, constant pressure is used
Dropping funel will dissolved with 400mg compounds Trp (Boc)-Gly-Asn (Trt)-Val-Leu-D-Ala 200mL DCM it is slow
It is added drop-wise in eggplant type bottle, nitrogen protection, after being added dropwise, eggplant type bottle is taken out from ice water, 25 DEG C of reactions are overnight (12h).With rotation
Turn evaporimeter to extract reaction solution out, oil pump drains extra DMF.Using methanol as eluant, eluent, with sephadex column (GE
Healthcare Sephadex LH-20) separation, the solvent portions containing polypeptide are collected, compound Huan [ is obtained after being spin-dried for;-Trp
(Boc)-Gly-Asn(Trt)-Val-Leu-D-Ala-]。
4) by compound Huan [-Trp(Boc)-Gly-Asn(Trt)-Val-Leu-D-Ala-]It is placed in 10mL centrifuge tubes,
The deprotecting regent of 5mL, under conditions of 0 DEG C, oscillating reactions 2h is added.Centrifugal sedimentation 3 times is carried out with the ether of 3 times of volumes,
Nitrogen dries up, and obtained white powder compound is further purified with high performance liquid chromatography, obtains final product Desotamide
B-3 (as shown in formula (I-3)), yield 52%.
Embodiment 4
1) first the DCM of 15mL is used to impregnate 2- chlorine trityl chloride resin 1000mg, (degree of substitution 1.03mmol/ at 30 DEG C
G) 0.5h pumps DCM.
The connection of single amino acids is reacted：Before connecing first amino acid, the Fmoc-D-Leu-OH amino of 335.0mg is weighed
Acid impregnates 1h with DIPEA the and 10mL DCM of 977 μ L and is activated, then meets reaction 1.5h at 30 DEG C of resin and obtain compound
Fmoc-D-Leu-2- chlorine trityl chloride resins.With the dichloromethane solution of 20% methanol volume ratios of 15mL, 30min is reacted,
It closes in site to having neither part nor lot in reaction on resin.After reaction, it is washed 3 times with DCM and DMF respectively, pumps cleaning solution.
The DMF solution of 25% piperidines volume ratios of 15mL is added into reaction tube, reacts 30min, removes Fmoc protecting groups.
The step of then above-mentioned connection amino acid removes the Fmoc protecting groups of end and then detects is repeated, is sequentially ingressed into
1000mg Fmoc-Val-OH, 1600mg Fmoc-Asn (Trt)-OH, 1000mg Fmoc-Gly-OH, 1500mg Fmoc-Trp
(Boc)-OH, 900mg Fmoc-Ala-OH obtain compound Ala-Trp (Boc)-Gly-Asn (Trt)-Val-D-Leu-2- chlorine
Trityl chloride resin.
2) Ala-Trp (Boc)-Gly-Asn (Trt)-Val-D-Leu-2- chlorine trityl chloride resins are transferred to 50mL
In centrifuge tube, it is added the cutting liquid of 15mL, under conditions of 0 DEG C, oscillating reactions 2.5h, then filtrate is filtered into eggplant type bottle.With
Rotary Evaporators remove the DCM in filtrate, and oil pump drains remaining liq, and 2mL toluene is added, then drain to get to yellowish toner
Last shape compound Ala-Trp (Boc)-Gly-Asn (Trt)-Val-D-Leu.
3) it weighs after HOAT 272.1mg are dissolved in 2mL DMF and is transferred in eggplant type bottle, then to weigh PyAOP 1042.3mg molten
It in the DCM of 200mL, is transferred in eggplant type bottle, adds 446.9 μ L of NMM.Eggplant type bottle is placed in 0 DEG C of ice water, constant pressure is used
Dropping funel will dissolved with 400mg compounds Ala-Trp (Boc)-Gly-Asn (Trt)-Val-D-Leu 200mL DCM it is slow
It is added drop-wise in eggplant type bottle, nitrogen protection, after being added dropwise, eggplant type bottle is taken out from ice water, 25 DEG C of reactions are overnight (12h).With rotation
Turn evaporimeter to extract reaction solution out, oil pump drains extra DMF.Using methanol as eluant, eluent, with sephadex column (GE
Healthcare Sephadex LH-20) separation, the solvent portions containing polypeptide are collected, compound Huan [ is obtained after being spin-dried for;-
Ala-Trp(Boc)-Gly-Asn(Trt)-Val-D-Leu-]。
4) by compound Huan [-Ala-Trp(Boc)-Gly-Asn(Trt)-Val-D-Leu-]It is placed in 10mL centrifuge tubes,
The deprotecting regent of 5mL, under conditions of 0 DEG C, oscillating reactions 2h is added.Centrifugal sedimentation 3 times is carried out with the ether of 3 times of volumes,
Nitrogen dries up, and obtained white powder compound is further purified with high performance liquid chromatography, obtains final product Desotamide
B-4 (as shown in formula (I-4)), yield 55%.
Embodiment 5
1) first the DCM of 15mL is used to impregnate 2- chlorine trityl chloride resin 1000mg, (degree of substitution 1.03mmol/ at 30 DEG C
G) 0.5h pumps DCM.
The connection of single amino acids is reacted：Before connecing first amino acid, the Fmoc-D-Leu-OH amino of 335.0mg is weighed
Acid impregnates 1h with DIPEA the and 10mL DCM of 977 μ L and is activated, then meets reaction 1h at 35 DEG C of resin and obtain compound Fmoc-
D-Leu-2- chlorine trityl chloride resins.With the dichloromethane solution of 20% methanol volume ratios of 15mL, 30min is reacted, to resin
On have neither part nor lot in the site of reaction and closed.After reaction, it is washed 3 times with DCM and DMF respectively, pumps cleaning solution.To reaction
The DMF solution of 25% piperidines volume ratios of 15mL is added in pipe, reacts 30min, removes Fmoc protecting groups.
The step of then above-mentioned connection amino acid removes the Fmoc protecting groups of end and then detects is repeated, is sequentially ingressed into
1000mg Fmoc-Leu-OH, 900mg Fmoc-Ala-OH, 1000mg Fmoc-Gly-OH, 1600mg Fmoc-Asn (Trt)-
OH, 1000mg Fmoc-Val-OH obtain compound Val-Asn (Trt)-Gly-Ala-Leu-D-Leu-2- chlorine trityl chlorides
Resin.
2) intermediate Val-Asn (Trt)-Gly-Ala-Leu-D-Leu-2- chlorine trityl chloride resins are transferred to 50mL
In centrifuge tube, it is added the cutting liquid of 15mL, under conditions of 0 DEG C, oscillating reactions 2.5h, then filtrate is filtered into eggplant type bottle.With
Rotary Evaporators remove the DCM in filtrate, and oil pump drains remaining liq, and 2mL toluene is added, then drain to get to yellowish toner
Last shape compound Val-Asn (Trt)-Gly-Ala-Leu-D-Leu.
3) it weighs after HOAT 329.0mg are dissolved in 2mL DMF and is transferred in eggplant type bottle, then to weigh PyAOP 1260.2mg molten
It in the DCM of 200mL, is transferred in eggplant type bottle, adds 540.3 μ L of NMM.Eggplant type bottle is placed in 0 DEG C of ice water, constant pressure is used
Dropping funel will slowly be added dropwise dissolved with the 200mL DCM of 400mg compounds Val-Asn (Trt)-Gly-Ala-Leu-D-Leu
Into eggplant type bottle, nitrogen protection after being added dropwise, takes out eggplant type bottle, 25 DEG C of reactions are overnight (12h) from ice water.It is steamed with rotation
Hair instrument extracts reaction solution out, and oil pump drains extra DMF.Using methanol as eluant, eluent, with sephadex column (GE
Healthcare Sephadex LH-20) separation, the solvent portions containing polypeptide are collected, compound Huan [ is obtained after being spin-dried for;-
Val-Asn(Trt)-Gly-Ala-Leu-D-Leu-]。
4) by compound Huan [-Val-Asn(Trt)-Gly-Ala-Leu-D-Leu-]It is placed in 10mL centrifuge tubes, is added
The deprotecting regent of 5mL, under conditions of 0 DEG C, oscillating reactions 1h.Centrifugal sedimentation 3 times, nitrogen are carried out with the ether of 3 times of volumes
Drying, obtained white powder compound are further purified with high performance liquid chromatography, obtain final product Desotamide B-5
(as shown in formula (I-5)), yield 42%.
Embodiment 6
1) first the DCM of 15mL is used to impregnate 2- chlorine trityl chloride resin 1000mg, (degree of substitution 1.03mmol/ at 35 DEG C
G) 0.5h pumps DCM.
The connection of single amino acids is reacted：Before connecing first amino acid, the Fmoc-D-Leu-OH amino of 335.0mg is weighed
Acid impregnates 1h with DIPEA the and 10mL DCM of 977 μ L and is activated, then meets reaction 1h at 35 DEG C of resin and obtain compound Fmoc-
D-Leu-2- chlorine trityl chloride resins.With the dichloromethane solution of 20% methanol volume ratios of 15mL, 30min is reacted, to resin
On have neither part nor lot in the site of reaction and closed.After reaction, it is washed 3 times with DCM and DMF respectively, pumps cleaning solution.To reaction
The DMF solution of 25% piperidines volume ratios of 15mL is added in pipe, reacts 30min, removes Fmoc protecting groups.
The step of then above-mentioned connection amino acid removes the Fmoc protecting groups of end and then detects is repeated, is sequentially ingressed into
1000mg Fmoc-Leu-OH, 1500mg Fmoc-Trp (Boc)-OH, 900mg Fmoc-Ala-OH, 1600mg Fmoc-Asn
(Trt)-OH, 1000mg Fmoc-Val-OH obtain compound Val-Asn (Trt)-Ala-Trp (Boc)-Leu-D-Leu-2-
Chlorine trityl chloride resin.
2) compound Val-Asn (Trt)-Ala-Trp (Boc)-Leu-D-Leu-2- chlorine trityl chloride resins are shifted
Into 50mL centrifuge tubes, it is added the cutting liquid of 15mL, under conditions of 0 DEG C, oscillating reactions 2.5h, then filtrate is filtered to eggplant type bottle
In.The DCM in filtrate is removed with Rotary Evaporators, oil pump drains remaining liq, and 2mL toluene is added, then drains to get to yellowish
Color powdered compounds Val-Asn (Trt)-Ala-Trp (Boc)-Leu-D-Leu.
3) it weighs after HOAT 257.7mg are dissolved in 2mL DMF and is transferred in eggplant type bottle, then weigh PyAOP 987.0mg and be dissolved in
The DCM of 200mL is transferred in eggplant type bottle, adds 423.2 μ L of NMM.Eggplant type bottle is placed in 0 DEG C of ice water, is dripped with constant pressure
Liquid funnel will slowly be dripped dissolved with the 200mL DCM of 400mg compounds Val-Asn (Trt)-Ala-Trp (Boc)-Leu-D-Leu
It is added in eggplant type bottle, nitrogen protection, after being added dropwise, eggplant type bottle is taken out from ice water, 25 DEG C of reactions are overnight (12h).With rotation
Evaporimeter extracts reaction solution out, and oil pump drains extra DMF.Using methanol as eluant, eluent, with sephadex column (GE
Healthcare Sephadex LH-20) separation, the solvent portions containing polypeptide are collected, compound Huan [ is obtained after being spin-dried for;-
Val-Asn(Trt)-Ala-Trp(Boc)-Leu-D-Leu-]。
4) by compound Huan [-Val-Asn(Trt)-Ala-Trp(Boc)-Leu-D-Leu-]It is placed in 10mL centrifuge tubes,
The deprotecting regent of 5mL, under conditions of 0 DEG C, oscillating reactions 2h is added.Centrifugal sedimentation 3 times is carried out with the ether of 3 times of volumes,
Nitrogen dries up, and obtained white powder compound is further purified with high performance liquid chromatography, obtains final product Desotamide
B-6 (as shown in formula (I-6)), yield 33%.
Detect example 1
The product of embodiment 1-6 is carried out to the detection of mass spectrum and nucleus magnetic hydrogen spectrum, testing result is shown in Fig. 2-13, can by figure
Know, structure and formula (I-1), formula (I-2), formula (I-3), formula (I-4), formula (I-5), the formula (I-6) of the product of embodiment 1-6 are one
It causes.
Application examples 1
The product of embodiment 1-6 is dissolved in dimethyl sulfoxide (DMSO) respectively, is configured to the liquid of 3.2mmol/L, -80 DEG C of preservations
For use, 320 μm of ol/L are diluted to fluid nutrient medium again before testing.Vancomycin is dissolved in sterile water, 1 μ is made into
The liquid of mol/L, -80 DEG C of preservations are for use.
The 200 μ L fluid nutrient mediums as blank control are added in 96 orifice plate, 1 hole, the training of 100 μ L MH liquid is added in the holes 3-11
Base is supported, 2 holes add the liquid of bacteria suspension 180 μ L and 20 μ L.100 μ L to 3 holes are taken from 2 holes, 3 holes take 100 μ L to 4 holes, sequentially into 10
Hole, 10 holes discard 100 μ L, and each hole drug concentration is respectively 32,16,8,4,2,1,0.5,0.25,0.125 μm of ol/L at this time.11
Hole is not added with liquid to be measured, and as positive control, control drug is vancomycin.96 orifice plates are cultivated in 37 DEG C of constant incubators
For 24 hours, OD values are measured, it is antibacterial for minimum less than 50% lowest concentration of drug with OD values with the OD values of Positive control wells for 100%
Concentration, concrete outcome are shown in Figure 14.
In Figure 14, NCTC7466 refers to streptococcus pneumonia (Streptococcus pneumoniae NCTC 7466),
ATCC 29213 refers to that staphylococcus aureus (Staphylococcus aureus ATCC 29213), ATCC 6051 refer to
Be bacillus subtilis (Bacillus subtilis ATCC 6051), ATCC 6633 refers to bacillus subtilis
(Bacillus subtilis ATCC 6633), De B-1 refer to that Desotamide B-1, De B-2 are referred to
Desotamide B-2, De B-3 refer to that Desotamide B-3, De B-4 refer to that Desotamide B-4, De B-5 refer to
Be Desotamide B-5, De B-6 refer to that Desotamide B-6, De B refer to Desotamide B；Pass through Figure 14
Understand that the compound fungistatic effect that De B-1 are synthesized relative to other is notable, it is possible to be developed into the elder generation of resisting gram-positive bacteria
Lead compound.
The preferred embodiment of the present invention has been described above in detail, still, during present invention is not limited to the embodiments described above
Detail can carry out a variety of simple variants to technical scheme of the present invention within the scope of the technical concept of the present invention, this
A little simple variants all belong to the scope of protection of the present invention.
It is further to note that specific technical features described in the above specific embodiments, in not lance
In the case of shield, can be combined by any suitable means, in order to avoid unnecessary repetition, the present invention to it is various can
The combination of energy no longer separately illustrates.
In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally
The thought of invention, it should also be regarded as the disclosure of the present invention.
Claims (10)
1. a kind of antibacterial cyclic peptides, which is characterized in that the structure of the antibacterial cyclic peptides such as formula (I-1), formula (I-2), formula (I-3), formula
(I-4), shown in any one of formula (I-5), formula (I-6),
2. antibacterial cyclic peptides according to claim 1, wherein shown in the structure of the antibacterial cyclic peptides such as formula (I-1).
3. a kind of preparation method of antibacterial cyclic peptides as described in claim 1, which is characterized in that including：
1) matrix resin is soaked in organic solvent, then removes solvent；Then by matrix resin successively with a variety of amino acid
It carries out multiple step-by-step processing and obtains the second resin；The step-by-step processing is：By resin, amino acid, N, N- diisopropylethylamine has
Solvent carries out haptoreaction, then carries out Seal treatment, washing, removing Fmoc protecting groups, washing；
2) the second resin, cutting liquid are subjected to oscillating reactions, then remove solvent and obtains straight chain peptide chain；
3) in the presence of protecting gas, by the straight chain peptide chain, 1- hydroxyl -7- azos benzotriazole, (3H-1,2,3- triazoles
Bing [4,5-b]Pyridine -3- oxygroups) three -1- pyrrolidinyls hexafluorophosphates, N-methylmorpholine, organic solvent carry out ring-closure reaction；
Then solvent is removed, then by post-processing primary cyclic peptide is made；
4) the primary cyclic peptide and deprotecting regent are subjected to haptoreaction to remove Boc protecting groups, then by system carry out from
Heart sedimentation, drying are to be made the antibacterial cyclic peptides；
Wherein, shown in the antibacterial cyclic peptides such as formula (I-1) in the case of, the amino acid of step-by-step processing is independent successively in step 2)
For Fmoc-Ala-OH, Fmoc-Val-OH, Fmoc-D-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp (Boc)-OH, Fmoc-
Gly-OH；In the case of shown in the antibacterial cyclic peptides such as formula (I-2), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Leu-OH、Fmoc-Leu-OH、Fmoc-Trp(Boc)-OH、Fmoc-Gly-OH、Fmoc-Asn(Trt)-OH、Fmoc-
Ala-OH；In the case of shown in the antibacterial cyclic peptides such as formula (I-3), the amino acid of step-by-step processing is stood alone as successively in step 2)
Fmoc-D-Ala-OH、Fmoc-Leu-OH、Fmoc-Val-OH、Fmoc-Asn(Trt)-OH、Fmoc-Gly-OH、Fmoc-Trp
(Boc)-OH；In the case of shown in the antibacterial cyclic peptides such as formula (I-4), the amino acid of step-by-step processing is independent successively in step 2)
For Fmoc-D-Leu-OH, Fmoc-Val-OH, Fmoc-Asn (Trt)-OH, Fmoc-Gly-OH, Fmoc-Trp (Boc)-OH,
Fmoc-Ala-OH；The amino acid of step-by-step processing is successively in the case of shown in the antibacterial cyclic peptides such as formula (I-5), in step 2)
Stand alone as Fmoc-D-Leu-OH, Fmoc-Leu-OH, Fmoc-Ala-OH, Fmoc-Gly-OH, Fmoc-Asn (Trt)-OH,
Fmoc-Val-OH；The amino acid of step-by-step processing is successively in the case of shown in the antibacterial cyclic peptides such as formula (I-6), in step 2)
Stand alone as Fmoc-D-Leu-OH, Fmoc-Leu-OH, Fmoc-Trp (Boc)-OH, Fmoc-Ala-OH, Fmoc-Asn (Trt)-
OH、Fmoc-Val-OH。
4. preparation method according to claim 2, wherein in step 1), in soaking process, described matrix resin,
The amount ratio of organic solvent is 1g：10-20mL；
Preferably, the immersion meets the following conditions：Soaking temperature is 15-35 DEG C, soaking time 30-50min.
5. preparation method according to claim 2, wherein in step 1), on the basis of 1g described matrix resins, every
It walks in step-by-step processing, the dosage of the n,N-diisopropylethylamine is 0.6-0.8g, the dosage of the organic solvent is 10-
20mL；
In the case of shown in the antibacterial cyclic peptides such as formula (I-1), the dosage of the Fmoc-Ala-OH is 290-300mg, described
The dosage of Fmoc-Val-OH is 330-1000mg, and the dosage of the Fmoc-D-Leu-OH is 330-1000mg, the Fmoc-
The dosage of Leu-OH is 330-1000mg, and the dosage of Fmoc-Trp (the Boc)-OH is 500-1500mg, the Fmoc-Gly-
The dosage of OH is 330-1000mg；
In the case of shown in the antibacterial cyclic peptides such as formula (I-2), the dosage of the Fmoc-D-Leu-OH is 330-340mg, institute
The dosage for stating Fmoc-Leu-OH is 330-1000mg, and the dosage of Fmoc-Trp (the Boc)-OH is 500-1500mg, described
The dosage of Fmoc-Gly-OH is 330-1000mg, and the dosage of Fmoc-Asn (the Trt)-OH is 600-1800mg, described
The dosage of Fmoc-Ala-OH is 300-900mg；
In the case of shown in the antibacterial cyclic peptides such as formula (I-3), the dosage of the Fmoc-D-Ala-OH is 290-300mg, institute
The dosage for stating Fmoc-Leu-OH is 330-1000mg, and the dosage of the Fmoc-Val-OH is 330-1000mg, the Fmoc-
The dosage of Asn (Trt)-OH is 600-1800mg, and the dosage of the Fmoc-Gly-OH is 330-1000mg, the Fmoc-Trp
(Boc) dosage of-OH is 500-1500mg；
In the case of shown in the antibacterial cyclic peptides such as formula (I-4), the dosage of the Fmoc-D-Leu-OH is 330-340mg, institute
The dosage for stating Fmoc-Val-OH is 330-1000mg, and the dosage of Fmoc-Asn (the Trt)-OH is 600-1800mg, described
The dosage of Fmoc-Gly-OH is 330-1000mg, and the dosage of Fmoc-Trp (the Boc)-OH is 500-1500mg, described
The dosage of Fmoc-Ala-OH is 300-900mg；
In the case of shown in the antibacterial cyclic peptides such as formula (I-5), the dosage of the Fmoc-D-Leu-OH is 330-340mg, institute
The dosage for stating Fmoc-Leu-OH is 330-1000mg, and the dosage of the Fmoc-Ala-OH is 300-900mg, the Fmoc-
The dosage of Gly-OH is 330-1000mg, and the dosage of Fmoc-Asn (the Trt)-OH is 600-1800mg, the Fmoc-Val-
The dosage of OH is 330-1000mg；
In the case of shown in the antibacterial cyclic peptides such as formula (I-6), the dosage of the Fmoc-D-Leu-OH is 330-340mg, institute
The dosage for stating Fmoc-Leu-OH is 330-1000mg, and the dosage of Fmoc-Trp (the Boc)-OH is 500-1500mg, described
The dosage of Fmoc-Ala-OH is 300-900mg, and the dosage of Fmoc-Asn (the Trt)-OH is 600-1800mg, the Fmoc-
The dosage of Val-OH is 330-1000mg；
Preferably, in every step step-by-step processing, the haptoreaction meets the following conditions：Reaction temperature is 15-35 DEG C, when reaction
Between be 1-1.5h；
It is highly preferred that the Seal treatment meets the following conditions：Treatment temperature is 30-35 DEG C, processing time 30-50min；
It is further preferred that on the basis of 1g described matrix resins, the Seal treatment use the volume ratio of addition 10-20mL for
1：The methanol and dichloromethane mixed solution of 4-5 carries out；
It is further preferred that on the basis of 1g described matrix resins, the step of the removing Fmoc protecting groups in using addition
The volume ratio of 10-20mL is 1：The piperidines and N,N-dimethylformamide mixed solution of 3-4 carries out.
6. preparation method according to claim 2, wherein in step 2), with second made from 1g described matrix resins
On the basis of resin, the dosage of the cutting liquid is 15-20mL；
Preferably, the oscillating reactions meets the following conditions：Reaction temperature is -5~0 DEG C, reaction time 2-4h；
It is highly preferred that the cutting liquid is 1 by volume ratio：0.8-1.2：Trifluoroethanol, glacial acetic acid and the dichloromethane group of 7-9
At.
7. preparation method according to claim 2, wherein in step 3), using the straight chain peptide chain of 400mg as base
The dosage of standard, the 1- hydroxyls -7- azo benzotriazole is 355-365mg, (3H-1,2, the 3- triazols;4,5-b]Pyrrole
Pyridine -3- oxygroups) dosages of three -1- pyrrolidinyl hexafluorophosphates is 1370-1380mg, the dosage of the N-methylmorpholine is
The dosage of 535-540mg, the organic solvent are 400-500mL；
Preferably, in step 3), the ring-closure reaction meets the following conditions：Reaction temperature is 15-35 DEG C, and the reaction time is
12-24h；
It is highly preferred that the post-processing is：Using methanol as eluant, eluent, by sephadex post separation, eluant, eluent is finally removed；
It is further preferred that the protection gas is selected from least one of nitrogen, helium and argon gas；
It is further preferred that before the ring-closure reaction, carried out in the environment for making an addition to -5~0 DEG C of material.
8. preparation method according to claim 2, wherein in step 4), made from the straight chain peptide chain with 400mg
On the basis of the primary cyclic peptide, the dosage of the deprotecting regent is 5-8mL；
Preferably, the haptoreaction meets the following conditions：Reaction temperature is -5~0 DEG C, reaction time 1.5-2h；
It is highly preferred that the deprotecting regent is 82-83 by volume ratio：4-6：4-6：4-6：The trifluoroacetic acid of 2-3, phenol, water,
Thioanisole and ethylene dithiol ether composition.
9. according to the preparation method described in any one of claim 3-8, wherein the organic solvent be selected from dichloromethane,
At least one of N,N-dimethylformamide, dimethyl sulfoxide (DMSO) and N-Methyl pyrrolidone；
Preferably, the reagent that uses of washing is dichloromethane and/or n,N-Dimethylformamide, in each washing operation
Washing times are 3-5 times；
It is highly preferred that described matrix resin is in 2- chlorine trityl chloride resin, Wang resins, MBHA resin and rink resins
At least one.
10. a kind of application of antibacterial cyclic peptides as described in claim 1 in antibacterial.
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