CN108250265A - A kind of new process for synthesizing the polypeptide containing intramolecular disulfide bond - Google Patents
A kind of new process for synthesizing the polypeptide containing intramolecular disulfide bond Download PDFInfo
- Publication number
- CN108250265A CN108250265A CN201611242844.2A CN201611242844A CN108250265A CN 108250265 A CN108250265 A CN 108250265A CN 201611242844 A CN201611242844 A CN 201611242844A CN 108250265 A CN108250265 A CN 108250265A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- disulfide bond
- oxidation
- intramolecular disulfide
- purified
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/20—Partition-, reverse-phase or hydrophobic interaction chromatography
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/02—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length in solution
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/06—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents
- C07K1/061—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using protecting groups or activating agents using protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the synthetic methods of chemiluminescent polypeptide synthesis technical field, the more particularly to polypeptide containing intramolecular disulfide bond.The method includes a kind of strategy directly aoxidized to the reversed-phase high performance liquid chromatography purified components of linear polypeptide crude product, and in this strategy, to polypeptide, linear crude product is purified by reversed-phase high performance liquid chromatography first, and obtained purified components are directly aoxidized.Polypeptide solution is aoxidized by being purified after concentration again by reversed-phase high performance liquid chromatography, it is final it is freeze-dried after obtain the sterling of target polypeptides.The method effectively prevents interference of certain impurity to oxidation product purity in the linear crude product of polypeptide, can effectively improve the purity and comprehensive yield of final product.Production procedure is simplified compared with conventional synthesis process, avoids freeze-drying repeatedly and the course of dissolution of polypeptide, while preferably protection polypeptide products quality, improves the feasibility of intramolecular disulfide bond polypeptide large-scale production.
Description
Technical field
The present invention is particularly suitable for linear about a kind of new method for preparing the polypeptide containing intramolecular disulfide bond in sequence
Peptide systhesis is ineffective, peptide sequence higher to final products purity requirement and having large-scale production demand and with this
The product that method obtains.
Background technology
Polypeptide be by a variety of amino acid according to certain a kind of compound for putting in order and being formed by peptide linkage, point
Minor structure is the bioactive substance for being related to various cell functions in organism between amino acid and protein.Polypeptide
It is fully synthetic that not only there is critically important theory significance, but also with important application value.It can be verified by the way that polypeptide is fully synthetic
The structure of one new polypeptide；New polypeptide is designed, for research structure and the relationship of function；For polypeptide biosynthesis reaction machine
System provides important information；It establishes model enzyme and synthesizes new polypeptide drugs etc..It is also, raw from animal, plant and microorganism etc.
Separation and Extraction polypeptide pole is limited by source in object, and the chemical synthesis of polypeptide then has brighter industrial prospect.In recent years
Hot spot to study polypeptide is had been transferred in the synthesis and biological assessment of cyclic peptide, and is very square by disulfide bond formation cyclic peptide
Just a kind of approach,.In many important extracellular polypeptides and protein molecule as included hormone, enzyme, growth factor, toxin, exempting from
Epidemic disease globulin, disulfide bond play vital effect in terms of molecular folding and rock-steady structure.Moreover, it is artificially induced one
A little disulfide bond can further constrain the conformation of polypeptide or protein, can reach and improve its bioactivity or thermodynamics
The purpose of stability.Also there is relatively unambiguous specific conformation just because of this kind of macromolecular cyclic peptide, so they can be more preferable
Ground carries out selective binding with receptor, and metabolic stability and bioavilability are significantly larger than linear peptides.In view of above disulfide bond
These advantages, the chemical synthesis of the polypeptide containing disulfide bond causes the broad interest of scientist always for a long time.
For disulfide bond in peptide molecule, the factor such as amino acid between polypeptide chain length, disulfide bond in terms of conformation is residual
Type and number of base etc. are even more important on the complexity influence of disulfide bond formation；And the selection of production technology, in synthesizing
Selection of cysteine (Cys) side chain thiol blocking group, oxidizing condition and purifying process etc., also to the life of disulfide bond polypeptide
Producing efficiency influences significantly.In common synthesis technology, mainly using trityl (Trt) or acetyl aminomethyl (Acm) as half
The side chain thiol blocking group of cystine synthesizes linear peptides by traditional Fmoc/tBu solid-phase synthesis, then through iodine (I2) or
The sulfydryl of two cysteines, generates intramolecular disulfide bond and obtains target cyclic peptide on dimethyl sulfoxide (DMSO) oxidation peptide chain.Line
Property peptide cyclization belong to inner molecular reaction, linear intermolecular reaction generation or cricoid dimerization and polymer occurs to avoid, generally
In the solution (10 of high dilution-3~10-4Mol/L it is carried out in).Also there is cyclic report in solid phase in recent years, that is, complete
It carries out being oxidized to ring directly on resin without cutting after the synthesis of entire polypeptide chain, what is utilized is that solid phase material is " false dilute
Release " it acts on avoiding intermolecular dimerization and poly side reaction, this dominance of strategies is not needing to the solution in high dilution
In aoxidized, and excessive oxidising agent and certain small molecule by-products can be by washing with being filtered to remove.It is but this
In strategy, when being cut the selection of cutting reagent can be limited to the presence of disulfide bond in polypeptide, lead to some amino acid residues
In the presence of the risk that side reaction occurs, so that thick peptide purity reduces, the purity of purifying yield and final product is most influenced at last.
So in polypeptide production, it is still one of main method for preparing disulfide bond polypeptide that polypeptide carries out oxidation in the solution.
In addition, reversed-phase high performance liquid chromatography (RP-HPLC) purifying process selection of polypeptide is also very crucial, mainly utilize
Retention time difference is detached to obtain target polypeptides molecule different peptide molecules in the chromatography column.Disulfide bond in polypeptide
The variation that polypeptide conformation variation can be caused so as to cause the hydrophilic and hydrophobic of polypeptide is formed, that is, polypeptide is in the chromatography column before and after aoxidizing
Retention time can have differences, and high-efficient liquid phase chromatogram purification method is using this species diversity come the polypeptide of separating reducing state and oxidation state.
And when directly being aoxidized and purified using the linear crude product of polypeptide, the hydrophilic and hydrophobic of certain impurity molecules may be with oxidation in thick peptide
Relatively, this will seriously affect the purification effect of oxidation polypeptide to polypeptide, and purifying yield is caused to substantially reduce or even in certain poles
Certain impurity molecules can not be efficiently separated out in the case of end, as depicted in figs. 1 and 2.In this case it is carried out using polypeptide sterling
Oxidation just becomes very necessary.However, in traditional sterling oxidation strategy, mainly comprise the steps of (see Fig. 3)：I) to not
It aoxidizes linear thick peptide product and first carries out RP-HPLC purifying, pass through freeze-drying removal solvent after collection respective components；Ii) will be more
Peptide freeze-dried powder is re-dissolved to be scattered in enough weak solutions and be aoxidized, thereafter will oxidation solution freeze-drying concentration；Iii) again into
Row RP-HPLC is purified, and final disulfide bond polypeptide products are obtained by third time freeze-drying.It can be seen that, in the method by
Quantitative demand is needed when polypeptide aoxidizes to concentrate requirement there are multiple freeze-drying and re-dissolve process, it is more numerous
It is trivial, time-consuming.
In addition, it is necessary to it is emphasized that freeze-drying operation repeatedly can also damage the quality of polypeptide products in itself
It is very big：The freezing stress generated in refrigerating process has destruction to biologically active polypeptide, it is considered that mainly by machine
Tool effect and solute effect cause；And the drying stress that drying process generates also has polypeptide drug destruction, may lead
Cause active peptides inactivation.
One object of the present invention is exactly the synthesis technology of simplified intermolecular disulfide bond polypeptide, is avoiding polypeptide products as possible
Under conditions of being freeze-dried repeatedly, the production efficiency of this kind of polypeptide products is improved.The further object of the present invention is to improve linearly
Crude product purity, with it is final provide have higher purity (>98%) it target polypeptides product and improves the comprehensive of these products and receives
Rate.
Invention content
The present invention prepares the new synthesis strategy of intramolecular disulfide bond polypeptide about a kind of simplicity, is particularly suitable for linear polypeptide
Synthetic effect it is bad and to product the higher peptide sequence of final purity requirement and the product obtained in this way.This side
Method includes a kind of strategy directly aoxidized to the reversed-phase high performance liquid chromatography purified components of linear polypeptide crude product, in this plan
In slightly, to polypeptide, linear crude product is purified by reversed-phase high performance liquid chromatography first, and obtained purified components directly carry out oxygen
Change.It is purified after oxidation polypeptide solution concentration again by reversed-phase high performance liquid chromatography, target polypeptides is obtained after freeze-drying
Sterling.The method effectively prevents interference of certain impurity to oxidation product purity in the linear crude product of polypeptide, can effectively improve
The purity and comprehensive yield of final product.Production procedure is simplified compared with conventional synthesis process, avoids the anti-of polypeptide
Multiple freeze-drying and course of dissolution while preferably protection polypeptide products quality, improve the big rule of intramolecular disulfide bond polypeptide
The feasibility of mould production.
Such method of the preparation containing intramolecular disulfide bond polypeptide is as shown in figure 3,1) a kind of prepare containing intramolecular disulfide
The method of key polypeptide, which is characterized in that this method includes the following steps successively：A) it is purified using rp-hplc more
The linear crude product of peptide isolates impurity molecule to improve Purity；B) the reversed-phase high performance liquid chromatography purifying group that will be collected into
Divide and directly aoxidized；C) condensing peptide oxidation solution after the completion of aoxidizing, it is pure to reuse rp-hplc progress
Change, the freeze-dried powder of target disulfide bond polypeptide sterling is obtained after freeze-drying.2) the method is characterized by the polypeptide oxidations of use
Method includes but are not limited to iodine oxidation, dimethylsulfoxide oxidation, hydrogen peroxide oxidation, air oxidation.3) the method is characterized by,
The method used when being concentrated to polypeptide solution after oxidation includes but are not limited to low-temperature rotary evaporation, low-temperature vacuum drying.4) originally
Method is characterized in that：A pair of of intramolecular disulfide bond is included but are not limited in peptide sequence.
Description of the drawings
Fig. 1 is the front and rear analytic type RP-HPLC detection figures of 1 crude product of polypeptide oxidation.It can be seen that, after oxidation target polypeptides with
The retention time of a certain impurity is very close in crude product before oxidation (being respectively 9.2 minutes and 9.3 minutes).
Fig. 2 is that polypeptide 1 obtains the analytic type RP-HPLC detection figures of product using conventional crude peptide direct oxidation method.After oxidation
Target polypeptides coexist with the impurity in crude product before oxidation, and further improving purity will become extremely difficult, and yield will also drop significantly
It is low.
Fig. 3 is to prepare showing for the production technology that the traditional processing technology containing intramolecular disulfide bond polypeptide and this patent describe
It is intended to.
Fig. 4 is the analytic type RP-HPLC detection figures of 1 sterling of polypeptide prepared by the new method of this patent description.It can be seen that
The purity of final products is significantly enhanced, up to more than 99%.Compared with conventional method, whole yield is also accordingly carried
It is high.Mobile phase is water and the mixed system of acetonitrile, and Detection wavelength is 220 nanometers, and flow velocity is 1 ml/min.
Fig. 5 is the analytic type RP-HPLC detection figures of 2 sterling of polypeptide prepared by the new method of this patent description, and purity reaches
98.21%.Mobile phase is water and the mixed system of acetonitrile, and Detection wavelength is 220 nanometers, and flow velocity is 1 ml/min.
Fig. 6 is the analytic type RP-HPLC detection figures of 3 sterling of polypeptide prepared by the new method of this patent description, and purity reaches
99.46%.Mobile phase is water and the mixed system of acetonitrile, and Detection wavelength is 220 nanometers, and flow velocity is 1 ml/min.
Fig. 7 is the analytic type RP-HPLC detection figures of 4 sterling of polypeptide prepared by the new method of this patent description, and purity reaches
97.52%.Mobile phase is water and the mixed system of acetonitrile, and Detection wavelength is 220 nanometers, and flow velocity is 1 ml/min.
Specific embodiment
Using manual solid-phase Fmoc/tBu methods, synthesized from c-terminus to aminoterminal direction：With 25 volume % hexahydropyridine/bis-
Methylformamide impregnates resin makes aminoterminal become free amino group, first time duration 8 to remove the Fmoc protecting groups of amino twice
Minute, second duration 10 minutes.Then, with 3 times of equivalents of amino acid/I-hydroxybenzotriazole (HOBt)/N, N'- diisopropyls
Carbodiimide (DIC) carries out grafting with resin and introduces second amino acid residue of c-terminus.It is so sequentially connected each ammonia repeatedly
Base acid residue is to complete the synthesis of whole polypeptide.It all needs alternately to be washed with dimethylformamide/dichloromethane after often step reaction above
It washs resin each four times, and all reaction is monitored by ninhydrin detection, if the reaction of some amino acid condensation is incomplete,
It repeats to be condensed once, reacts the full sequence that the sequence obtained after the completion is target polypeptides.
Use cutting reagent (trifluoroacetic acid：1,2- dithioglycols：Thioanisole：Phenol：Water：Tri isopropyl silane=
68.5:10:10:5:3.5:1, v/v) target polypeptides from resin are cleaved and remove Side chain protective group and (cut 3 at 30 DEG C
Hour).Filtrate, which is added in 10 times of cold anhydrous ethers of amount, makes polypeptide Precipitation, centrifugation removal ether.It is washed with ether more
Peptide crude product is three times, dry after centrifuging respectively.
Polypeptide crude product is purified first using preparative RP-HPLC：Chromatograph packing material is 10 μm of reverse phase C18 silica gel,
Aperture isColumn size is 50 × 250mm.Mobile phase A be containing 0.05 volume % trifluoroacetic acids, 2 volume % acetonitriles
Aqueous solution, Mobile phase B are 90 volume % acetonitrile solutions, and flow velocity is 25 milliliters per minute, 220 nanometers of ultraviolet detection wavelength.
Oxidant (oxidant is measured again based on polypeptide crude product before purification) is added in into the purified components of collection, uses analytic type
RP-HPLC tracks oxidation process, is concentrated oxidation solution by freeze-drying after oxidation.Again by RP-HPLC pairs of preparative
It is purified, and is collected after respective components are freeze-dried up to the target polypeptides sterling of soft fluffy state.Its chemical constitution is by matrix
Assisted laser desorption ionisation flight time mass spectrum (MALDI-TOF-MS) is characterized, and its purity is then by analytic type RP-HPLC
(Venusil C18-10 × 250mm, flow velocity：1 ml/min, Detection wavelength：220 nanometers) it provides.
Present invention provides the polypeptide products obtained with method provided by the invention.It can see, use from these products
This method has obtained the disulfide bond peptide molecule of high-purity.From yield, sterling purity or technique ease
For, the method is all that conventional synthetic methods are incomparable.These examples are provided to illustrate this patent, and in any way all
It is not intended that limitation of the scope of the invention.
Embodiment 1
The sequence of polypeptide 1 is H-Cys-Asn-AA1-Gly-Ser-AA2-Cys-OH.It is prepared by solid phase Fmoc/tBu methods
The linear crude product of polypeptide 1 purifies this crude product by preparative RP-HPLC.Ice vinegar is added in into the purified components of collection
After acid makes its acetic acid ratio reach 20 volume %, iodine/acetum that 0.2M is slowly added dropwise under stirring is aoxidized (by before purification
Polypeptide crude product meter, 5eq iodine molecules), a length of 60 minutes or so during reaction.Oxidation process is tracked with analytic type RP-HPLC, it is linear more
After the peak of peptide completely disappears in spectrogram, vitamin c solution is slowly added dropwise into reaction solution, until the brown of oxidation solution is complete
It takes off.Solution will be aoxidized by low-temperature vacuum drying to carry out after concentrating in right amount, it be carried out again by preparative RP-HPLC pure
Change, collect respective components, be freeze-dried after removal solvent up to puffy, 1 sterling of polypeptide of high-purity.It is direct with conventional crude product
Method of purification is compared, and the purity of final products is greatly improved, and comprehensive yield also makes moderate progress；And it is aoxidized with traditional sterling
Strategy is compared, and freeze-drying repeatedly is avoided in new method and is re-dissolved, the quality of final polypeptide products is made also to obtain
Preferably protection, also, technique is simplified in whole process, and the production cycle greatly shortens.The result pair of attached drawing 2 and attached drawing 4
Ratio illustrates, improvement of the application method compared to conventional method on purity, yield.
Embodiment 2
The sequence of polypeptide 2 is H-Cys-AA3-Thr-Leu-AA4-Ser-Ala-Gly-AA5-Leu-Leu-Gly-Cys-
His-Ala-Val-NH2.The linear crude product of polypeptide 2 is prepared by solid phase Fmoc/tBu methods, it is thick to this by preparative RP-HPLC
Product are purified.Pure acetic acid is added in into the purified components of collection makes its acetic acid ratio reach 20 volume %, stirs lower slowly drop
Iodine/acetum of 0.2M is added to be aoxidized (based on polypeptide crude product before purification, 5eq iodine molecules), a length of 60 minutes left sides during reaction
It is right.Oxidation process is tracked with analytic type RP-HPLC, after the absorption peak of linear polypeptide completely disappears in spectrogram, to reaction solution
In vitamin c solution is slowly added dropwise, until oxidation solution brown take off completely.Oxidation solution is carried out by low-temperature rotary evaporation
After appropriate concentration, it is purified again by preparative RP-HPLC, respective components are collected, after freeze-drying removes solvent
Up to puffy, 2 sterling of polypeptide of high-purity.
Embodiment 3
The sequence of polypeptide 3 is H-Asn-AA6-Thr-Cys-AA7-Ala-Met-Leu-AA8-Leu-Lys-Gly-Cys-
Gln-NH2.The linear crude product of polypeptide 3 is prepared by solid phase Fmoc/tBu methods, is purified by preparative RP-HPLC.To receipts
Pure acetic acid is added in the purified components of collection makes its acetic acid ratio reach 20 volume %, and iodine/acetic acid of 0.2M is slowly added dropwise under stirring
Solution is aoxidized (based on polypeptide crude product before purification, 5eq iodine molecules), is reacted 60 minutes or so.It is tracked with analytic type RP-HPLC
After the absorption peak of linear polypeptide completely disappears in spectrogram, vitamin c solution is slowly added dropwise into reaction solution for oxidation process,
Brown to oxidation solution is taken off completely.Oxidation solution is carried out after concentrating in right amount by low-temperature rotary evaporation, again by system
Standby type RP-HPLC purifies it, collects respective components, after freeze-drying removal solvent up to puffy, high-purity it is more
3 sterling of peptide.
Embodiment 4
The sequence of polypeptide 4 is H-Lys-AA9-Cys-Phe-AA10-Val-Cys-AA11-AA12-Gly-Ile-Cys-
Tyr-AA13-Arg-Cys-Arg-NH2(Cys3-Cys16；Cys7-Cys12).Polypeptide sequence is completed by solid phase Fmoc/tBu methods
Using Fmoc-Cys (Acm)-OH at the synthesis of row, wherein Cys7 and Cys12, the linear crude product of polypeptide 4 is obtained after cutting, wherein
Cys7 and Cys12 is with Acm protecting groups, and Cys3 and Cys16 side chains are free sulfhydryl group.Pass through preparative RP-HPLC first
Linear crude product is purified, the DMSO of 20 volume % is added in into the purified components of collection, is reacted under being stirred at room temperature, is used
Analytic type RP-HPLC tracks this oxidation process, after the absorption peak of linear polypeptide completely disappears in spectrogram, passes through cryogenic vacuum
It is dry to be concentrated oxidation solution in right amount, it is purified again by preparative RP-HPLC.Add into the purified components of collection
After entering glacial acetic acid its acetic acid ratio being made to reach 20 volume %, iodine/acetum that 0.2M is slowly added dropwise under stirring is aoxidized and (is pressed
Polypeptide crude product meter before purification, 5eq iodine molecules), a length of 60 minutes or so during reaction.With mass spectrum and analytic type RP-HPLC tracking oxygen
After change process, the complete removal of Acm protecting groups namely monocyclic peptide molecule completely disappear, vitamin is slowly added dropwise into reaction solution
C solution, until the brown of oxidation solution is taken off completely.Solution will be aoxidized by low-temperature vacuum drying to carry out after concentrating in right amount, again
It is purified by preparative RP-HPLC, collects respective components, up to puffy, high-purity after freeze-drying removal solvent
4 sterling of polypeptide containing two pairs of intramolecular disulfide bonds.
Claims (4)
1. a kind of prepare the method containing intramolecular disulfide bond polypeptide, which is characterized in that this method includes the following steps successively：
A) using the linear crude product of rp-hplc purified polypeptide, impurity molecule is isolated to improve Purity；
B) the reversed-phase high performance liquid chromatography purified components being collected into directly are aoxidized；
C) condensing peptide oxidation solution, reuses rp-hplc and is purified after the completion of aoxidizing, and is freeze-dried
The freeze-dried powder of target disulfide bond polypeptide sterling is obtained afterwards.
2. the method containing intramolecular disulfide bond polypeptide is prepared as described in claim 1, it is characterised in that：The polypeptide oxygen of use
Change method includes but are not limited to iodine oxidation, dimethylsulfoxide oxidation, hydrogen peroxide oxidation, air oxidation.
3. the method containing intramolecular disulfide bond polypeptide is prepared as described in claim 1, it is characterised in that：To polypeptide after oxidation
The method that solution uses when concentrating includes but are not limited to low-temperature rotary evaporation, low-temperature vacuum drying.
4. the method containing intramolecular disulfide bond polypeptide is prepared as described in claim 1, it is characterised in that：It is wrapped in peptide sequence
It includes but is not limited only to a pair of of intramolecular disulfide bond.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611242844.2A CN108250265A (en) | 2016-12-29 | 2016-12-29 | A kind of new process for synthesizing the polypeptide containing intramolecular disulfide bond |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611242844.2A CN108250265A (en) | 2016-12-29 | 2016-12-29 | A kind of new process for synthesizing the polypeptide containing intramolecular disulfide bond |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108250265A true CN108250265A (en) | 2018-07-06 |
Family
ID=62719766
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611242844.2A Pending CN108250265A (en) | 2016-12-29 | 2016-12-29 | A kind of new process for synthesizing the polypeptide containing intramolecular disulfide bond |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108250265A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020077781A1 (en) * | 2018-10-16 | 2020-04-23 | 深圳翰宇药业股份有限公司 | Preparation method for synthesis of polypeptide containing two pairs of disulfide bonds and kit thereof, and method for preparing plecanatide |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005100381A2 (en) * | 2004-04-08 | 2005-10-27 | Millennium Pharmaceuticals, Inc. | Processes for preparing eptifibatide and pertinent intermediate compounds |
| WO2011011342A1 (en) * | 2009-07-20 | 2011-01-27 | Mallinckrodt Inc. | Synthesis of desmopressin |
| CN104974237A (en) * | 2015-07-18 | 2015-10-14 | 济南康和医药科技有限公司 | Solid-phase synthesis method of ziconotide by segment process |
| CN106167514A (en) * | 2016-08-29 | 2016-11-30 | 杭州湃肽生化科技有限公司 | The synthesis of a kind of Linaclotide and purification process |
-
2016
- 2016-12-29 CN CN201611242844.2A patent/CN108250265A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005100381A2 (en) * | 2004-04-08 | 2005-10-27 | Millennium Pharmaceuticals, Inc. | Processes for preparing eptifibatide and pertinent intermediate compounds |
| WO2011011342A1 (en) * | 2009-07-20 | 2011-01-27 | Mallinckrodt Inc. | Synthesis of desmopressin |
| CN104974237A (en) * | 2015-07-18 | 2015-10-14 | 济南康和医药科技有限公司 | Solid-phase synthesis method of ziconotide by segment process |
| CN106167514A (en) * | 2016-08-29 | 2016-11-30 | 杭州湃肽生化科技有限公司 | The synthesis of a kind of Linaclotide and purification process |
Non-Patent Citations (4)
| Title |
|---|
| PASCAL VERDIE,ET AL.: "Oxyfold: A Simple and Efficient Solid-Supported Reagent for Disulfide Bond Formation", 《CHEM. ASIAN J.》 * |
| 徐仲等: "奥曲肽固相合成及环化的研究", 《哈尔滨工业大学学报》 * |
| 曹本文等: "选择性氧化形成三对二硫键合成齐考诺肽", 《合成化学》 * |
| 王巧娥等: "《甘草深加工技术》", 31 January 2011, 中国轻工业出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020077781A1 (en) * | 2018-10-16 | 2020-04-23 | 深圳翰宇药业股份有限公司 | Preparation method for synthesis of polypeptide containing two pairs of disulfide bonds and kit thereof, and method for preparing plecanatide |
| CN111057129A (en) * | 2018-10-16 | 2020-04-24 | 深圳翰宇药业股份有限公司 | Preparation method and kit for synthesizing polypeptide containing two pairs of disulfide bonds, and preparation method of polycaprolactam |
| CN111057129B (en) * | 2018-10-16 | 2024-01-16 | 深圳翰宇药业股份有限公司 | Preparation method and kit for synthesizing polypeptide containing two pairs of disulfide bonds, and preparation method of pramipexole |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101463072B (en) | Preparation of cholecystokinin octapeptide | |
| CN105001298B (en) | A kind of synthesis isolation and purification method of indissoluble polypeptide | |
| CN103012563A (en) | Solid-phase synthesis method of antibacterial peptide Iseganan | |
| CN105504012A (en) | Preparation method of polypeptide | |
| CN103694316A (en) | Preparation method of argireline | |
| CN106243194B (en) | A kind of method of improved Fmoc Solid-phase synthesis peptides oxytocin | |
| CN108250265A (en) | A kind of new process for synthesizing the polypeptide containing intramolecular disulfide bond | |
| CN103709233B (en) | A kind of method of Fmoc method Solid Phase Synthesis Thymopentin Using | |
| CN104163853A (en) | Method for preparing linaclotide | |
| CN104211801A (en) | Method for preparing lixisenatide | |
| JP4942295B2 (en) | Method for producing L-alanine-L-glutamine | |
| CN101846649B (en) | Phosphorylated and/or glycosylated protein or peptide one-step enrichment modification determination method | |
| CN106967149A (en) | A kind of preparation method of the cysteine polypeptide of palmitoylation modification | |
| CN103897029A (en) | Preparation method for romidepsin | |
| Andrei et al. | Purification and partial characterization of an antiviral active peptide from Melia azedarach L. | |
| CN103740797A (en) | Method for preparing high-hydrolysis degree functional oligopeptide by use of high-temperature peanut meal | |
| CN102775471A (en) | Method for synthesizing cholecystokinin octapeptide by combining solid phase method and liquid phase method | |
| CN103450346B (en) | Liquid-phase synthesis method of eptifibatide | |
| CN102336813B (en) | A kind of preparation method of synthesizing proteidin with solid phase polypeptide | |
| CN106749497A (en) | Lasso trick polypeptide it is chemical fully synthetic | |
| CN111187343B (en) | Peony 2S albumin and extraction method and application thereof | |
| CN113214376A (en) | Novel method for synthesizing centipede toxin RhTX and spider toxin GsMTx4 | |
| CN104558159B (en) | The solid phase synthesis process of Angiomax intermediate | |
| CN107417786B (en) | Preparation method of thymosin alpha 1 | |
| CN104478994A (en) | Fullerene single-addition line peptide as well as preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |