CN108148115A - A kind of cyclic peptide new synthetic method and its application in drug development - Google Patents
A kind of cyclic peptide new synthetic method and its application in drug development Download PDFInfo
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- CN108148115A CN108148115A CN201810104326.7A CN201810104326A CN108148115A CN 108148115 A CN108148115 A CN 108148115A CN 201810104326 A CN201810104326 A CN 201810104326A CN 108148115 A CN108148115 A CN 108148115A
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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
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
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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
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- 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
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Abstract
The invention discloses a kind of cyclic peptide new synthetic method and its applications in drug development.Cyclic peptide improves stability, is the available strategy and key technology of polypeptide drug structure optimization because its special structure can effectively resist enzyme degradation.The synthesis mode of current cyclic peptide includes the cyclization of natural amino acid and the cyclization by small molecule auxiliary etc..The natural cyclisation method stability of natural amino acid is poor, chemical synthesis is difficult.By the cyclization technology of small molecule auxiliary because the introducing of small molecule can influence the structure of cyclic peptide and lead to some side effects.The present invention effectively combines both the above synthesis strategy, reacting based on maleimide and sulfydryl, it has first synthesized side-chain amino group and cyclic peptide is obtained by the reaction by the amine-modified lysine of maleimide, then with cysteine sulfydryl, the strategy reaction condition is mild, high selectivity, the complete yield height of rapid reaction.
Description
Technical field
The invention belongs to cyclic peptide to synthesize field, and in particular to a kind of cyclic peptide new synthetic method and its answering in drug development
With.
Background technology
Polypeptide is to be present in a kind of combination of amino acids with extensive physiological activity and abundant structure diversity in organism
Object, pharmacological activity is strong, high selectivity, and antigenicity is low, is not easy to accumulate in vivo, fewer with the interaction of other drugs,
Toxicity is weak, therefore with very high drug development potentiality.It is existing at present bent including cyclosporin, Epothilones, Caspofungin, Austria
Numerous polypeptide drug listings such as peptide, are widely used in the treatment of AIDS, tumour and infectious diseases etc..It is more from structure
Peptide can be divided into linear polypeptide and cyclic peptide, and linear polypeptide makes its stability poor due to the enzyme degradation being easily widely present in vivo, and half
Declining, the phase is short, and leading to it, there are lot of challenges in new drug development.Cyclic peptide can effectively resist enzyme because of its special structure in contrast
Degradation, so as to improve stability, therefore is the available strategy and key technology of polypeptide drug structure optimization by polypeptide cyclisation.
According to the difference of cyclization mode, cyclic peptide can be divided into the cyclization of natural amino acid and the cyclization by small molecule auxiliary etc.
Two ways.The natural cyclization of natural amino acid includes on polypeptide chain：Polypeptide N-terminal and C-terminal are with amido bond cyclization, amino acid side chain
Between the modes such as cyclization, the cyclization between N-terminal and amino acid side chain and the cyclization between C-terminal and amino acid side chain, these are
The major way of coupling reagent cyclization, the major defect of this cyclization mode is stability, and poor (such as disulfide bond is easily by internal paddy
The reductive conditions such as the sweet peptide of Guang destroy) and carboxyl, amino etc. cause chemical synthesis difficult since reactivity is low.By small molecule
The polypeptide cyclization technology of auxiliary includes：The modes such as click chemistry, olefin metathesis, the nucleophilic displacement of fluorine of nitrine-alkynes, these cyclization sides
The major defect of formula is that the introducing of small molecule can influence the structure of cyclic peptide and cause some unpredictable to a certain extent
Side effect is introduced into the use of the factors such as heavy metal such as the copper catalyst in nitrine-alkynes ring-closure reaction, can lead in addition in building-up process
Synthetic product heavy-metal residual is caused so as to influence drug quality.New cyclic peptide synthetic technology needs further to be developed, by above two
It is a kind of very promising developing direction that kind synthesis strategy, which combines,.
Michael addition reaction between maleimide and sulfydryl, since reaction condition is mild, high selectivity, reacts fast
Fast yield completely is high, is widely used to peptide modified and derivatization at present.In view of lysine natural abundance is high and side chain contains
Long chain amino, therefore available maleimide modifies its side-chain amino group, lysine can be under certain conditions
Cysteine sulfydryl reaction is cyclic and obtains cyclic peptide.Above-mentioned synthesis strategy is effectively combined when first two cyclic peptide synthesis strategy, is led to
Cross effectively reduces small numerator modified influence to lysine side-chain progress modified with functional group, additionally due to numerous amino acid sulfydryls spread out
Exploitation (the Synthetic cysteine surrogates used in native chemical ligation [J] of biology
.Mol Biosyst, 2013,9 (5)：826-833), the use scope of this method has greatly been expanded, in addition reaction condition temperature
With, therefore be a kind of very promising polypeptide synthesis method.
Invention content
The purpose of the present invention is to provide a kind of cyclic peptide new synthetic method and its application in drug development, this method gram
Deficiency of the prior art is taken, reaction condition is mild, yield is high, easy purification.
The present invention is achieved through the following technical solutions above-mentioned purpose：
A kind of cyclic peptide new synthetic method and its application in drug development, method include the following steps：
(1), using resin as carrier, by be based on the Solid-phase synthesis peptides technology of 9-fluorenylmethyloxycarbonyl (Fmoc) from C-terminal to
N-terminal is condensed successively containing lysine (Lys, derivative or the like) and cysteine (Cys, derivative or the like)
Peptide sequence obtains the resin with reference to polypeptide；Wherein, lysine is repaiied using the side-chain amino group of new design synthesis by maleimide
Fmoc-Lys (maleimido)-OH of decorations, the sulfydryl of cysteine use acetyl aminomethyl (Acm) orthogonal protection.
(2), sulfhydryl protected base (Aom) in the polypeptide resin that step (1) obtains is sloughed, obtains the exposed polypeptide tree of sulfydryl
Fat, washing.
(3), the polypeptide resin for obtaining step (2) carries out solid phase cyclization reaction, washing.
(4), the cyclic peptide resin for obtaining step (3) carries out cleavage reaction, and cyclic peptide is made to be broken from resin, and cutting liquid carries out
Polypeptide precipitation, centrifugation, purifying, freeze-drying, obtain target cyclic peptide.
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, polypeptide sequence described in step (1)
Row are to choose KRGDSfC as template peptide, and Wang resin is carrier, verifies each step reaction condition and result.
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, Fmoc described in step (1) are protected
Protecting amino acid, sequence is followed successively by cysteine (Cys), phenylalanine (Phe), serine (Ser), asparagus fern from C-terminal to N-terminal condensation
Amide (Asp), glycine (Gly), arginine (Arg), lysine (Lys), setting-up point are room temperature, condensation reaction time
For 1h.
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, Fmoc- described in step (1)
The synthetic method of Lys (Maleimido)-OH is：By lysine and N- methoxycarbonylmaleimides with the ratio of molar ratio 1: 3
0 DEG C, in saturated sodium bicarbonate solution is added to, reaction time 80min.Through isolating and purifying to obtain Fmoc- after reaction completion
Lys(Maleimido)-OH。
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, first described in step (1)
The method of amino acid and resin-bonded is：Wang resin (0.25mmol) is weighed in Peptide systhesis pipe, add methylene chloride submergence, quiet
After putting swelling 30min, it is spare to filter out solvent.In addition Fmoc-Cys (Acm)-OH (5 times, 1.25mmol) is weighed in conical flask,
Adding in 0 DEG C of dichloromethane makes its dissolving, adds I-hydroxybenzotriazole (5 times, 1.25mmol) and N, N- diisopropyl carbon
Diimine (5 times, 1.25mmol), ice bath stirring 30min carry out activation of amino acid (dichloromethane or ultrasound is added dropwise if any precipitation).
In the resin that the amino acid activated addition has been swollen, add 4-dimethylaminopyridine (0.5 times, 0.13mmol), seal polypeptide
Composite tube acutely vibrates, and reacts at room temperature 10-15h.After the completion of reaction, reaction solution is filtered off, with n,N-Dimethylformamide, dichloro
Each four washings resin of methane alternating, overnight vacuum is dry or dries, and weighs and calculates degree of substitution.
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, obtain described in step (1)
Fmoc-Cys (Acm)-wang resin need to be blocked, and first be placed in obtained Fmoc-Cys (Acm)-wang resin
Resin sealing end, sealing are carried out in (10% n,N-diisopropylethylamine, 20% aceticanhydride, 70%N, dinethylformamide) solution
Reaction vessel, room temperature reaction 0.5h (or 1.25mmol acetic anhydride, 0.25mmol pyridines, dichloromethane react 1h).Reaction is completed
Afterwards, resin is washed, is stayed with spare.
A kind of cyclic peptide new synthetic method of the present invention, described in step (1) in addition to cysteine, remaining amino acid uses
Following method synthesis：N is configured, dinethylformamide: piperidines=(4: 1) solution is added to Fmoc-Cys (Acm)-wang
Oscillating reactions 5min and 15min in resin, it is each primary, slough Fmoc groups.Then with n,N-Dimethylformamide (4mL ×
10s × 3 time), dichloromethane (4mL × 10s × 3 time), n,N-Dimethylformamide (4mL × 10s × 3 time) alternately washing tree
Fat.Then Fmoc- amino acid (4 times, 1mmol) is weighed, benzotriazole-N, N, N ', N '-tetramethylurea hexafluorophosphate
(HBTU) (3.8 times, 0.95mmol), I-hydroxybenzotriazole (3.8 times, 0.95mmol), n,N-diisopropylethylamine (8 times,
3min 2mmol) is reacted at room temperature in conical flask, adds in resin, reacts at room temperature 1h.Followed by with n,N-Dimethylformamide (4mL
× 10s × 3 time), dichloromethane (4mL × 10s × 3 time), n,N-Dimethylformamide (4mL × 10 s × 3 time) washing resin.
Deprotection, washing, condensation, washing are repeated, until required each amino acid is reacted in order to terminating.Obtain Lys
(Maleimido)-Arg(Mtr)-Gly-Asp(OtBu)-Ser(tBu)-Phe-Cys(Acm)-wang resin。
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, the polypeptide obtained in step (1)
Resin in order to verify the correctness of synthetic peptide sequence, takes a small amount of polypeptide resin to crack, and cleavage method is：(trifluoroacetic acid is configured
: phenol: water: tri isopropyl silane=88: 5: 5: 2) solution 10mL, oscillating reactions 2h filter out reaction solution, and with a small amount of trifluoro second
Acid repeats to wash 3 times, merges reaction solution and is concentrated in vacuo (absorption tower oil pump vacuumizes or nitrogen blowing is to less than 5ml), can also add
(2800r × 4min) is centrifuged after ice ether precipitation, precipitation is taken, is dissolved in acetonitrile-water (1: 1) solution, obtain linear polypeptide Lys
(Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys(Acm)。
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, sulfydryl described in step (2) are protected
The removal methods of shield base Acm are：The polypeptide that step (2) obtains is added to acetonitrile-water (1: 1), containing the molten of 0.1% trifluoroacetic acid
In liquid, it is 20 times of silver acetates measured of polypeptide to add, and a small amount of three (2- carboxyethyls) phosphine is added dropwise after having reacted, reaction temperature is room
Temperature, reaction time 2h, purifying obtain sulfydryl unprotected Lys (Maleimido)-Arg (Mtr)-Gly-Asp (OtBu)-Ser
(tBu)- Phe-Cys-wang resin。
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, sulfhydryl protected base in step (2)
The polypeptide resin obtained on a small quantity product is taken to verify its correctness after the removing of Acm, cracks to obtain linear peptides by above-mentioned cleavage method
Lys(Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys。
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, polypeptide ring described in step (3)
Change method is：The polypeptide resin that step (2) obtains is added in buffer solution (50mM 4- hydroxyethyl piperazineethanesulfonic acids, 5mM second
Ethylenediamine tetraacetic acid (EDTA), pH 7.4), reaction temperature is room temperature, and reaction time 5h is cracked simultaneously after the completion of reaction by above-mentioned cleavage method
Purifying obtains cyclic peptide sterling, cyclo [Lys-Arg-Gly-Asp-Ser-Phe-Cys].
A kind of cyclic peptide new synthetic method of the present invention and its application in drug development, the analysis of each step products therefrom and
Purification process boils method using RP-HPLC color, using analysis liquid phase, chooses C18, 4.6 μ m 250mm pillars, using preparation
Liquid phase chooses C18, the purifying of 15 μ m 250mm pillars.
Beneficial effects of the present invention：
Cyclic peptide analog is finally made in a kind of cyclic peptide new synthetic method of the present invention and its application in drug development
Purity is 91.7%, yield 54.1%, and yield, which has, to be greatly improved, and each step reaction mild condition.
Description of the drawings
Attached drawing 1 is the high-efficient liquid phase chromatogram of Fmoc-Lys (Maleimido)-OH.
Attached drawing 2 is the mass spectrogram of Fmoc-Lys (Maleimido)-OH.
The hydrogen that attached drawing 3 is Fmoc-Lys (Maleimido)-OH is composed.
The carbon that attached drawing 4 is Fmoc-Lys (Maleimido)-OH is composed.
Attached drawing 5 is the efficient liquid phase of linear polypeptide Lys (Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys (Acm)
Chromatogram.
Attached drawing 6 is the mass spectrogram of linear polypeptide Lys (Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys (Acm).
Attached drawing 7 is the high performance liquid chromatography of linear polypeptide Lys (Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys
Figure.
Attached drawing 8 is the mass spectrogram of linear polypeptide Lys (Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys.
Attached drawing 9 is the high-efficient liquid phase chromatogram of cyclic peptide cyclo [Lys-Arg-Gly-Asp-Ser-Phe-Cys].
Attached drawing 10 is the mass spectrogram of cyclic peptide cyclo [Lys-Arg-Gly-Asp-Ser-Phe-Cys].
Specific embodiment
Following non-limiting examples can make those of ordinary skill in the art be more fully understood the present invention, but not with
Any mode limits the present invention.
Claims and abbreviation meaning used in the description are listed in the table below：
| Wang resin | 4- (methylol) Phenoxymethyl polystyrene resin |
| Acm | Acetyl aminomethyl |
| tBu | Tertiary butyl |
| OtBu | Tert-butoxy |
| Mtr | 4- methoxyl groups -2,3,6- trimethylphenysulfonyls |
| HBTU | Benzotriazole tetramethyl tetrafluoro boric acid |
The synthesis of embodiment 1cyclo [Lys-Arg-Gly-Asp-Ser-Phe-Cys]
(1) preparation of Fmoc-Lys (Maleimido)-OH：Be configured saturated sodium bicarbonate solution 200ml in round-bottomed flask simultaneously
0 DEG C of ice bath is put in spare.Weigh (2mmol, 0.74g) Fmoc-Lys-OH, (6mmol, 0.93g) N- methoxycarbonyl groups Malaysia acyl
Imines is added into round-bottomed flask 0 DEG C of ice bath in configured good 200ml saturated sodium bicarbonate solutions and reacts 80 min.
By efficiently preparing liquid phase purifying, obtained product purity is 89.6% twice after the completion of reaction, yield 56.4%.
(2) preparation of Fmoc-Cys (Acm)-wang resin：Wang resin degree of substitution used in the experiment is 0.48mmol/g,
It weighs (0.25mmol, 0.52g) Wang resin and is put into Peptide systhesis pipe the submergence that adds methylene chloride, filtered out after standing swelling 30min
Solvent is spare.Dichloromethane is placed in 0 DEG C of ice bath, and weighs Fmoc-Cys (Acm)-OH (5 times, 1.25mmol, 0.52g) and puts
In conical flask, adding in 0 DEG C of dichloromethane makes its dissolving, and add in I-hydroxybenzotriazole (5 times, 1.25mmol,
0.17g) and N, N- diisopropylcarbodiimide (5 times, 1.25mmol, 198 μ L), ice bath stirring 30min carry out activation of amino acid
(continuing that dichloromethane or ultrasound is added dropwise if any precipitation).In the resin that the amino acid activated addition has been swollen, 4- bis- is added in
Methylamino pyridine (0.5 times, 0.13mmol, 0.02g) seals Peptide systhesis pipe, reacts at room temperature 10-15 hours.After the completion of reaction,
Reaction solution is filtered off, n,N-Dimethylformamide (4 times), (4 times) washing resins of dichloromethane, overnight vacuum is dry or dries, and claims
Re-computation degree of substitution is 61.1%.
(3) preparation of Lys (Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys (Acm)-wang resin：First will
Obtained Fmoc-Cys (Acm)-wang resin resins are placed in Peptide systhesis reaction tube, and adding in 10ml, (10%N, N- bis- is different
Propylethylamine, 20% aceticanhydride, 70%N, dinethylformamide) in solution, sealed reaction vessel, room temperature reaction 30min is to tree
Fat is blocked.After the completion of reaction, with n,N-Dimethylformamide (4mL × 10s × 3 time), dichloromethane (4mL × 10s × 3
It is secondary), n,N-Dimethylformamide (4mL × 10s × 3 time) washing resin.After having washed, (20ml) N, N- dimethyl methyls are configured
Amide: piperidines=(4: 1) mixed solution, point front and rear be added to twice in Fmoc-Cys (Acm)-wang resin are reacted respectively
5min and 15 min, to slough Fmoc groups as far as possible.Then again with n,N-Dimethylformamide (4mL × 10s × 3 time),
Dichloromethane (4mL × 10s × 3 time), n,N-Dimethylformamide (4mL × 10s × 3 time) washing resin.Then Fmoc- is weighed
Phe-OH (4 times, 1mmol, 0.39g), HBTU (3.8 times, 0.95mmol, 0.36g), I-hydroxybenzotriazole (3.8 times,
0.95mmol, 0.13g), after n,N-diisopropylethylamine (8 times, 2mmol, 335 μ L) room temperature activates 3min, addition has been deprotected
Cys (Acm)-Wang resin resins in, in Peptide systhesis pipe acutely oscillation room temperature reaction 1 hour, and still use N, N-
Dimethylformamide (4mL × 10s × 3 time), dichloromethane (4mL × 10s × 3 time), n,N-Dimethylformamide (4mL × 10s
× 3 times) washing resin.Fmoc-Ser (tBu)-OH (4 times, 1mmol, 0.39g), Fmoc-Asp (OtBu)-OH (4 are weighed successively
Times, 1mmol, 0.42g), Fmoc-Gly-OH (4 times, 1mmol, 0.30g), Fmoc-Arg (Mtr)-OH (4 times, 1mmol,
0.61g), Fmoc-Lys (Maleimido)-OH (4 times, 1mmol, 0.45g), and repeat be deprotected, wash, being condensed, washing this
A little steps, until reaction terminates.
(4) preparation of Lys (Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys (Acm)：After Peptide systhesis,
Take a small amount of polypeptide resin, cracking verification synthetic product correctness.Configuration (trifluoroacetic acid: phenol: water: tri isopropyl silane=88:
5: 5: 2) solution 10mL, it adds in and has synthesized in the Peptide systhesis pipe of end, oscillating reactions 2h filters out reaction solution, and with a small amount of three
Fluoroacetic acid repeats to wash 3 times, merges reaction solution and is concentrated in vacuo (absorption tower oil pump vacuumizes or nitrogen blowing is to less than 5ml), also may be used
3-4 times (2800r × 4min) is centrifuged after being precipitated with ether on the rocks, takes precipitation, is dissolved with acetonitrile-water (1: 1) solution, is obtained linear
Polypeptide.It is again 92.4% through preparing liquid phase product after purification purity, reaction yield 47.2%
(5)Lys(Maleimido)-Arg(Mtr)-Gly-Asp(OtBu)-Ser(tBu)-Phe-Cys-wang resin
Preparation：The polypeptide that step (2) obtains is added to acetonitrile-water (1: 1), in the solution containing 0.1% trifluoroacetic acid, adding is
The silver acetates of 20 times of polypeptide amount, a small amount of three (2- carboxyethyls) phosphine of dropwise addition after react, reaction temperature is room temperature, and the reaction time is
2h obtains the unprotected peptide-wang resin containing sulfydryl.
(6) preparation of Lys (Maleimido)-Arg-Gly-Asp-Ser-Phe-Cys：Cysteine side chain thiol is protected
Base Acm takes a small amount of polypeptide resin to crack to obtain linear polypeptide by above-mentioned cleavage method after sloughing.It is obtained after purification through preparing liquid phase
Purity is 94.1%, yield 71.7%.
(7) preparation of cyclo [Lys-Arg-Gly-Asp-Ser-Phe-Cys]：The polypeptide resin that step (2) obtains is added
Enter into 50mL buffer solutions (50mM 4- hydroxyethyl piperazineethanesulfonic acids, 5Mm ethylenediamine tetra-acetic acids, pH 7.4), reaction temperature is
Room temperature, reaction time 5h are cracked, are purified and be lyophilized to obtain cyclized polypeptide sterling, purity 91.7%, and yield is
54.1%.
The present invention is any ripe preferred embodiment is disclosed as above Yi Shang content, but it is not limited to the present invention
The people of this technology is known, without departing from the spirit and scope of the present invention, can all do various change and modification, therefore the present invention
Protection domain should be subject to what claims were defined.
Claims (6)
1. a kind of cyclic peptide new synthetic method and its application in drug development, which is characterized in that include the following steps：
(1), it using resin as carrier, is condensed successively from C-terminal to N-terminal containing depending on ammonia by the Solid-phase synthesis peptides technology based on Fmoc
The peptide sequence of sour (Lys, derivative or the like) and cysteine (Cys, derivative or the like) obtain combining more
The resin of peptide；Wherein, the side-chain amino group that lysine is synthesized using new design is by the amine-modified Fmoc-Lys of maleimide
(maleimido)-OH, the sulfydryl of cysteine use acetyl aminomethyl (Acm) orthogonal protection；
(2), sulfhydryl protected base (Acm) in the polypeptide resin that step (1) obtains is sloughed, the exposed polypeptide resin of sulfydryl is obtained, washes
It washs；
(3), the polypeptide resin for obtaining step (2) carries out solid phase cyclization reaction, washing；
(4), the cyclic peptide resin for obtaining step (3) carries out cleavage reaction, and cyclic peptide is made to be broken from resin, and cutting liquid carries out polypeptide
Precipitation, centrifugation, purifying, freeze-drying, obtain target cyclic peptide；
2. according to claim 1, a kind of cyclic peptide new synthetic method and its application in drug development, it is characterised in that：
Peptide sequence described in step (1) contains lysine (Lys) and the cysteine (Cys) of acetyl aminomethyl protection.
3. according to claim 1, a kind of cyclic peptide new synthetic method and its application in drug development, it is characterised in that：
Step is not limited to lysine described in (1), also includes the derivative containing active amino, such as aminocaproic acid.
4. according to claim 1, a kind of cyclic peptide new synthetic method and its application in drug development, it is characterised in that：
Step is not limited to cysteine described in (1), also includes the amino acid derivativges containing sulfhydrylation, such as the dried meat of sulfhydrylation
Propylhomoserin.
5. according to claim 1, a kind of cyclic peptide new synthetic method and its application in drug development, it is characterised in that：
The synthetic method of Fmoc-Lys (maleimido)-OH described in step (1) is：By Fmoc-Lys-OH and N- methoxycarbonyl groups
Maleimide is added in alkaline solution, reacts certain time.
6. according to claim 1, a kind of cyclic peptide new synthetic method and its application in drug development, it is characterised in that：
Polypeptide cyclisation method is described in step (3)：The exposed polypeptide resin of sulfydryl that step (2) is obtained is added to buffer solution
In (4- hydroxyethyl piperazineethanesulfonic acids, ethylenediamine tetra-acetic acid), reaction a period of time.
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| CN112250734A (en) * | 2020-09-30 | 2021-01-22 | 南开大学 | Method for synthesizing cyclic peptide by using three components of lysine and tyrosine as anchor points through synergistic reaction |
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| CN114174313A (en) * | 2019-04-09 | 2022-03-11 | 芝加哥大学 | Macrocyclization and multimerization of peptides and proteins of the general type using diels-alder cycloaddition |
| CN114249801A (en) * | 2021-12-06 | 2022-03-29 | 南开大学 | Cyclopeptide compound and preparation method and application thereof |
| CN115368437A (en) * | 2022-06-21 | 2022-11-22 | 深圳市第二人民医院(深圳市转化医学研究院) | Method for solid-phase synthesis of cyclic polypeptide |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110092445A1 (en) * | 2008-07-25 | 2011-04-21 | Abbott Laboratories | Amyloid ß peptide analogues, oligomers thereof, processes for preparing and composi-tions comprising said analogues or oligomers, and their uses |
| CN103242441A (en) * | 2013-05-27 | 2013-08-14 | 江苏施美康药业股份有限公司 | Solid-phase synthesis method of Ziconotide |
-
2018
- 2018-01-30 CN CN201810104326.7A patent/CN108148115A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110092445A1 (en) * | 2008-07-25 | 2011-04-21 | Abbott Laboratories | Amyloid ß peptide analogues, oligomers thereof, processes for preparing and composi-tions comprising said analogues or oligomers, and their uses |
| CN103242441A (en) * | 2013-05-27 | 2013-08-14 | 江苏施美康药业股份有限公司 | Solid-phase synthesis method of Ziconotide |
Non-Patent Citations (1)
| Title |
|---|
| XAVIER ELDUQUE 等: "Straightforward Synthesis of Cyclic and Bicyclic Peptides", 《AMERICAN CHEMICAL SOCIETY》 * |
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| CN114174313A (en) * | 2019-04-09 | 2022-03-11 | 芝加哥大学 | Macrocyclization and multimerization of peptides and proteins of the general type using diels-alder cycloaddition |
| CN112500450A (en) * | 2019-09-16 | 2021-03-16 | 翰宇药业（武汉）有限公司 | Synthetic method of octreotide |
| CN112250734A (en) * | 2020-09-30 | 2021-01-22 | 南开大学 | Method for synthesizing cyclic peptide by using three components of lysine and tyrosine as anchor points through synergistic reaction |
| CN114249801A (en) * | 2021-12-06 | 2022-03-29 | 南开大学 | Cyclopeptide compound and preparation method and application thereof |
| CN115368437A (en) * | 2022-06-21 | 2022-11-22 | 深圳市第二人民医院(深圳市转化医学研究院) | Method for solid-phase synthesis of cyclic polypeptide |
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