CN111233714B - Preparation method of MAPS polypeptide - Google Patents
Preparation method of MAPS polypeptide Download PDFInfo
- Publication number
- CN111233714B CN111233714B CN202010190425.9A CN202010190425A CN111233714B CN 111233714 B CN111233714 B CN 111233714B CN 202010190425 A CN202010190425 A CN 202010190425A CN 111233714 B CN111233714 B CN 111233714B
- Authority
- CN
- China
- Prior art keywords
- fmoc
- polypeptide
- lys
- maps
- 8maps
- 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.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C271/00—Derivatives of carbamic acids, i.e. compounds containing any of the groups, the nitrogen atom not being part of nitro or nitroso groups
- C07C271/06—Esters of carbamic acids
- C07C271/08—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms
- C07C271/10—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms
- C07C271/22—Esters of carbamic acids having oxygen atoms of carbamate groups bound to acyclic carbon atoms with the nitrogen atoms of the carbamate groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of hydrocarbon radicals substituted by carboxyl groups
-
- 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
- C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
-
- 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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/02—Ortho- or ortho- and peri-condensed systems
- C07C2603/04—Ortho- or ortho- and peri-condensed systems containing three rings
- C07C2603/06—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members
- C07C2603/10—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings
- C07C2603/12—Ortho- or ortho- and peri-condensed systems containing three rings containing at least one ring with less than six ring members containing five-membered rings only one five-membered ring
- C07C2603/18—Fluorenes; Hydrogenated fluorenes
Abstract
The invention relates to the technical field of polypeptide synthesis, in particular to a preparation method of MAPS polypeptide, which comprises the following steps: synthesizing a novel MAPS polypeptide linker, namely DiFmoc-Lys (plus); DiFmoc-Lys (plus) was loaded onto the resin to give 8MAPS resin; synthesizing 8AMPS polypeptide crude product according to Fmoc solid phase synthesis method, and separating and purifying by HPLC to obtain 8MAPS polypeptide refined product. The invention has the beneficial effects that: the invention develops a novel unnatural amino acid DiFmoc-Lys (plus) on the basis of the existing Lys, so that two bifurcate arms of the DiFmoc-Lys (plus) are completely consistent in length and structure, the subsequent amino acid coupling reaction efficiency is completely consistent, and the crude product purity and the success rate of MAPS polypeptide are greatly improved.
Description
Technical Field
The invention relates to the technical field of polypeptide synthesis, in particular to a preparation method of a polypeptide antigen MAPS.
Background
Commonly used vaccines include live attenuated vaccines, inactivated vaccines and subunit vaccines. Most current subunit vaccines are produced using recombinant DNA technology using microorganisms or mammalian cells. However, antigens with smaller molecular weights are difficult to prepare by recombinant DNA techniques and chemical synthesis becomes a simpler approach. The synthetic peptide vaccine is a vaccine which is designed according to the amino acid sequence of an antigen and synthesized by a chemical method, has high safety, but has small molecular weight and weak immunogenicity, and only aims at a single epitope, so that the application of the synthetic peptide vaccine is limited. Although the immunogenicity of the synthetic peptide vaccine can be improved by coupling the vaccine with other carrier proteins, the effect is still not ideal because the influence of the vaccine on the structure of the target antigen peptide cannot be estimated due to the introduction of unrelated proteins. The branched multiple antigen peptide system generally refers to a system having several Lys (n) as a core, which utilizes the characteristic that two amino groups of lysine (Lys) can form peptide bonds with the carboxyl group of another amino acid. Connecting a plurality of same or different antigen peptides, and preparing the macromolecular branched polypeptide by a chemical synthesis method. Multiple Antigen Peptides (MAPS) were first proposed by James PT in 1988, and this system can solve the difficulty of weak immunogenicity of synthetic antigen peptide vaccines without introducing other proteins.
The preparation of MAP can be divided into a dispersion method and a concentration method. The former is directly started from the core and gradually derivatizes outwards to synthesize the polypeptide, and the method is generally realized by SPPS; the latter is an indirect, modular synthesis method. Peptide chains and templates are assembled together after separate synthetic purification to give the final product, a strategy that is usually accomplished by both SPPS and LPPS.
In the synthesis of MAPS polypeptide by a dispersion method, the biggest difficulty is that the reaction efficiency of each amino acid in subsequent coupling is uneven because the length of a branched arm connecting two amino groups on the common Di-Fmoc-Lys is inconsistent, the coupling efficiency of the amino acid coupled on the alpha amino group is low because the branched arm is short and the steric hindrance is large, so that the deletion of peptide is possible, the great difficulty is caused to the purification of the later-stage polypeptide, and the success rate of the MAP polypeptide is greatly reduced.
There is also a method of adding a beta alanine to the alpha amino group of Lys to obtain Di-Fmoc-Lys (beta-Ala), wherein Di-Fmoc-Lys (beta-Ala) has a uniform length of the branched arms, but Di-Fmoc-Lys (beta-Ala) is grafted with beta-alanine to introduce an amide bond, which can improve reaction uniformity to some extent, but the reaction efficiency on both arms is not completely uniform.
Therefore, there is a need to develop a synthetic method that can improve the purity and yield of crude MAPS polypeptides.
Disclosure of Invention
The first purpose of the invention is to overcome the defects of the prior art and provide a novel unnatural amino acid DiFmoc-Lys (plus), the structural formula is shown as follows:
in a second aspect, the present invention provides the use of the above novel unnatural amino acid DiFmoc-Lys (plus) for the synthesis of multiple antigen peptide MAPS.
The third aspect of the present invention provides a method for synthesizing the above novel unnatural amino acid DiFmoc-Lys (plus), comprising the following steps:
(1) fmoc-butanediamine in NaBr and HNO2Under the catalysis of the N-Fmoc-4-bromobutylamine;
(2) reacting the N-Fmoc-4-bromobutylamine with metal magnesium to obtain Fmoc-N-4-bromobutylamine Grignard reagent for later use;
(3)NH2-Lys (Fmoc) -Otbu in NaNO2Addition of I under catalysis2Is reacted toTo I-Lys (Fmoc) -Otbu;
(4) reacting N-Fmoc-4-bromobutylamine Grignard reagent with I-Lys (Fmoc) -Otbu to obtain 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid tert-butyl ester;
(5) removing tert-butyl ester from 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid by TFA acidolysis to obtain 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid, namely DiFmoc-Lys (plus); the specific reaction formula is as follows:
in a fourth aspect, the invention provides a method for preparing a MAPS polypeptide, comprising the steps of:
a. synthesizing a novel MAPS polypeptide linker, namely DiFmoc-Lys (plus);
b. DiFmoc-Lys (plus) was loaded onto the resin to give 8MAPS resin;
c. synthesizing 8AMPS polypeptide crude product according to Fmoc solid phase synthesis method, and separating and purifying by HPLC to obtain 8MAPS polypeptide refined product.
Preferably, the step b comprises the following steps: adding a proper amount of resin of the queen cell blank into a reactor, and preparing the 8AMPS resin according to the sequence of primary condensation → washing → hydroxyl sealing → washing → deprotection → washing → secondary condensation → washing → deprotection → washing → third condensation.
Preferably, DMF is taken as a solvent, and DiFmoc-Lys (plus), TBTU and DIEA in a molar ratio of 1:1:2 are added for reaction for 1-2 hours; the molar weight of each raw material added in the secondary condensation in the step b is 2 times that of the primary condensation, and the molar weight of each raw material added in the third condensation in the step b is 4 times that of the primary condensation.
Preferably, the hydroxyl-capping reagent used in step b is DMF: pyridine: acetic anhydride ═ 2:2:1(v/v), and the deprotection reagent is 20% piperidine/DMF solution.
Preferably, the specific conditions in step c are as follows: and (2) coupling each protected amino acid one by taking the prepared 8MAPS resin according to an Fmoc solid-phase synthesis method to obtain 8MAPS resin polypeptide, cutting, crystallizing and drying the 8MAPS resin polypeptide to obtain a crude 8MAPS polypeptide product, and separating, purifying and lyophilizing by HPLC to obtain a refined 8MAPS polypeptide product. The structure of 8MAPS resin polypeptides is shown below:
the invention has the beneficial effects that: the invention develops a novel unnatural amino acid DiFmoc-Lys (plus) on the basis of the existing Lys, so that two bifurcate arms of the DiFmoc-Lys (plus) are completely consistent in length and structure, the subsequent amino acid coupling reaction efficiency is completely consistent, and the crude product purity and the success rate of MAPS polypeptide are greatly improved.
Drawings
FIG. 1 is a mass spectrum of a top-quality MAPS polypeptide prepared in example 3;
FIG. 2 is a high performance liquid chromatogram of a competitive MAPS polypeptide prepared in example 3;
FIG. 3 is a mass spectrum of a top-quality MAPS polypeptide prepared in example 4;
FIG. 4 is a high performance liquid chromatogram of a competitive MAPS polypeptide prepared in example 4;
FIG. 5 is a mass spectrum of a top-quality MAPS polypeptide prepared in example 5;
FIG. 6 is a high performance liquid chromatogram of a competitive MAPS polypeptide prepared in example 5.
Detailed Description
The technical solutions of the present invention are described below by way of specific examples, but the scope of the present invention is not limited thereto.
Example 1 preparation of DiFmoc-Lys (plus)
1. Weighing 31g of Fmoc-butanediamine, placing the Fmoc-butanediamine in a 500ml flask, adding 12.3g of NaBr, dissolving in THF, slowly dropwise adding HNO2, magnetically stirring for reaction for 2 hours, and detecting the reaction end point by TLC to obtain 26.2g of N-Fmoc-4-bromobutylamine; then adding 2g of metal magnesium for reaction to obtain 19.8g of Fmoc-N-4-bromobutylamine Grignard reagent for later use;
2. weighing 42.5g of NH2-Lys (Fmoc) -Otbu, placing the weighed material into a 500ml flask, adding acetone to dissolve the material, adding 6.9g of NaNO2 and 15.2g I2, and stirring the mixture to react to obtain 37.5g of I-Lys (Fmoc) -Otbu;
3. taking 19.8g of N-Fmoc-4-bromobutylamine Grignard reagent in the step 1 and I-Lys (Fmoc) -Otbu37.5g in the step 2 to react to obtain 17.5g of 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid tert-butyl ester; the tert-butyl 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoate was subjected to acid hydrolysis with TFA to remove tert-butyl ester, yielding 15.6g of 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid, which was DiFmoc-Lys (plus). MW: 646.8.
EXAMPLE 28 preparation of AMPS resin
Weighing 10g of empty royal jelly resin, placing the empty royal jelly resin into a reactor, adding DMF to swell for 30 minutes, then adding 0.65g of DiFmoc-Lys (plus), 0.32g of TBTU and 1ml of DIEA, reacting for 2 hours, washing, then adding 100ml of hydroxyl sealing reagent, reacting for 1 hour, then adding 100ml of deprotection reagent, stirring and reacting for 30 minutes, then adding 1.3g of DiFmoc-Lys (plus), 0.64g of TBTU and 2ml of DIEA, and reacting for 1 hour; after deprotection, 2.6g of DiFmoc-Lys (plus), 1.28g of TBTU, and 4ml of DIEA were added, and the reaction was carried out for 1 hour, followed by washing and drying to obtain 12.5g of 8AMPS resin, which was used for a degree of resin substitution of 0.36mmol/g with a spectrophotometer.
EXAMPLE 3 preparation of polypeptide Fine products
Taking 2.78g of 8MAPS resin with the substitution degree of 0.36mmol/g and the branched polypeptide sequence of Trp-Gln-Pro-Pro-Arg-Ala-Arg-Ile, coupling each amino acid one by one according to Fmoc solid phase synthesis method, wherein the materials used in each condensation step are shown in Table 1:
TABLE 1
| Step (ii) of | Protected amino acids | Condensing agent |
| 1 | Fmoc-Ile-OH | DIC/HOBT/DIEA |
| 2 | Fmoc-Arg(pbf)-OH | DIC/HOBT/DIEA |
| 3 | Fmoc-Ala-OH | DIC/HOBT/DIEA |
| 4 | Fmoc-Arg(pbf)-OH | DIC/HOBT/DIEA |
| 5 | Fmoc-Pro-OH | DIC/HOBT/ |
| 6 | Fmoc-Pro-OH | DIC/HOBT/ |
| 7 | Fmoc-Gln(Trt)-OH | DIC/HOBT/ |
| 8 | Fmoc-Trp(Boc)-OH | DIC/HOBT/DIEA |
After coupling, the resin polypeptide is cut, crystallized and dried to obtain 9.1g of MAPS polypeptide crude product, the theoretical crude product amount is 0.36 x 2.78 x 9350 x 9.36g, and the crude product is separated and purified by HPLC to obtain 3.5g of MAPS polypeptide refined product with the purity of more than 95%, the purification yield is 38.5%, and the total yield is 37.4%. MW: 9350. the mass spectrum and HPLC of the MAPS polypeptide fine product prepared in example 3 are shown in FIGS. 1 and 2.
Example 4 preparation of polypeptide Fine products
Taking 2.78g of 8MAPS resin with the substitution degree of 0.36mmol/g and the branched polypeptide sequence of Lys-Lys-Leu-Glu-Lys-Lys-Thr-Thr, coupling each amino acid one by one according to Fmoc solid phase synthesis method, wherein the materials used in each step of condensation are shown in Table 2:
TABLE 2
| Step (ii) of | Protected amino acids | Condensing agent |
| 1 | Fmoc-Thr(tbu)-OH | DIC/ |
| 2 | Fmoc-Thr(tbu)-OH | DIC/HOBT/DIEA |
| 3 | Fmoc-Lys(Boc)-OH | DIC/HOBT/DIEA |
| 4 | Fmoc-Lys(Boc)-OH | DIC/HOBT/DIEA |
| 5 | Fmoc-Glu(Otbu)-OH | DIC/ |
| 6 | Fmoc-Leu-OH | DIC/ |
| 7 | Fmoc-Lys(Boc)-OH | DIC/ |
| 8 | Fmoc-Lys(Boc)-OH | DIC/HOBT/DIEA |
After coupling, the resin polypeptide is cut, crystallized and dried to obtain 8.6g of crude MAPS polypeptide, the theoretical crude quantity is 0.36 × 2.78 × 8966 × 8.97g, and the crude MAPS polypeptide is separated and purified by HPLC to obtain 3.2g of refined MAPS polypeptide with purity of more than 90%, the purification yield is 37.2% and the total yield is 35.6%. MW: 8966 mass spectra and HPLC profiles of the MAPS polypeptide top-quality prepared in example 4 are shown in FIGS. 3 and 4.
EXAMPLE 5 preparation of polypeptide Fine products
Taking 2.78g of 8MAPS resin with the degree of substitution of 0.36mmol/g and the branched polypeptide sequence of Cys-Ala-Gly-Lys-Ala-Ala-Gly-Arg-Asn-Gly, coupling each amino acid one by one according to Fmoc solid phase synthesis method, wherein the materials used in each step of condensation are shown in Table 3:
TABLE 3
| Step (ii) of | Protected amino acids | Condensing agent |
| 1 | Fmoc-Gly-OH | DIC/ |
| 2 | Fmoc-Asn(trt)-OH | DIC/HOBT/DIEA |
| 3 | Fmoc-Arg(pbf)-OH | DIC/HOBT/DIEA |
| 4 | Fmoc-Gly-OH | DIC/HOBT/DIEA |
| 5 | Fmoc-Ala-OH | DIC/ |
| 6 | Fmoc-Ala-OH | DIC/ |
| 7 | Fmoc-Lys(Boc)-OH | DIC/ |
| 8 | Fmoc-Gly-OH | DIC/ |
| 9 | Fmoc-Ala-OH | DIC/ |
| 10 | Fmoc-Cys(Trt)-OH | DIC/HOBT/DIEA |
After coupling, the resin polypeptide is cut, crystallized and dried to obtain 8.2g of MAPS polypeptide crude product, the theoretical crude product amount is 0.36 × 2.78 × 8396 × 8.40g, and the crude product is separated and purified by HPLC to obtain 3.2g of MAPS polypeptide refined product with purity of over 98%, the purification yield is 39%, and the total yield is 38.1%. MW: 8396, the mass spectrum and HPLC of the MAPS polypeptide fine product prepared in example 5 are shown in FIGS. 5 and 6.
In light of the foregoing description of the preferred embodiment of the present invention, many modifications and variations will be apparent to those skilled in the art without departing from the spirit and scope of the invention. The technical scope of the present invention is not limited to the content of the specification, and must be determined according to the scope of the claims.
Claims (8)
1. An amino acid DiFmoc-Lys (plus) having the formula:
2. use of the amino acid DiFmoc-lys (plus) according to claim 1 for the preparation of a multiple antigen peptide MAPS.
3. Process for the preparation of the amino acid DiFmoc-lys (plus) according to claim 1, characterized in that it comprises the following steps:
(1) fmoc-butanediamine in NaBr and HNO2Is catalyzed to obtainTo N-Fmoc-4-bromobutylamine;
(2) reacting the N-Fmoc-4-bromobutylamine with metal magnesium to obtain Fmoc-N-4-bromobutylamine Grignard reagent for later use;
(3)NH2-Lys (Fmoc) -Otbu in NaNO2Addition of I under catalysis2The reaction of (a) gives I-Lys (Fmoc) -Otbu;
(4) reacting N-Fmoc-4-bromobutylamine Grignard reagent with I-Lys (Fmoc) -Otbu to obtain 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid tert-butyl ester;
(5) removing tert-butyl ester from 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid by TFA acidolysis to obtain 2-N-Fmoc-butylamine-6-N-Fmoc-amino-hexanoic acid, namely DiFmoc-Lys (plus); the specific reaction formula is as follows:
4. a method for preparing a MAPS polypeptide, comprising the steps of:
a. synthesizing MAPS polypeptide linker, namely DiFmoc-Lys (plus); the structure is as follows:
b. DiFmoc-Lys (plus) was loaded onto the resin to give 8MAPS resin;
c. synthesizing a crude 8MAPS polypeptide product according to an Fmoc solid-phase synthesis method, and then separating and purifying by HPLC to obtain a refined 8MAPS polypeptide product; the structure of 8MAPS resin polypeptides is shown below:
5. the method of producing the MAPS polypeptide of claim 4, wherein step b comprises the steps of: adding a proper amount of the resin blank into a reactor, and preparing the 8MAPS resin according to the sequence of primary condensation → washing → hydroxyl sealing → washing → deprotection → washing → secondary condensation → washing → deprotection → washing → three times of condensation.
6. The method for preparing MAPS polypeptide according to claim 5, wherein the condition of the first condensation in step b is that DMF is used as solvent, DiFmoc-Lys (plus), O-benzotriazole-N, N, N ', N' -tetramethyluronium tetrafluoroborate and N, N-diisopropylethylamine are added in a molar ratio of 1:1:2 for reaction for 1-2 hours; the molar weight of each raw material added in the secondary condensation in the step b is 2 times that of the raw material added in the primary condensation, and the molar weight of each raw material added in the third condensation in the step b is 4 times that of the raw material added in the primary condensation.
7. The method of making MAPS polypeptides of claim 5, wherein the hydroxy-capping reagent used in step b is DMF: pyridine: acetic anhydride ═ 2:2:1, and the deprotection reagent is 20% piperidine/DMF solution.
8. The method of producing the MAPS polypeptide of claim 4, wherein the specific conditions in step c are as follows: and (2) coupling each protected amino acid one by taking the prepared 8MAPS resin according to an Fmoc solid-phase synthesis method to obtain 8MAPS resin polypeptide, cutting, crystallizing and drying the 8MAPS resin polypeptide to obtain a crude 8MAPS polypeptide product, and separating, purifying and lyophilizing by HPLC to obtain a refined 8MAPS polypeptide product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010190425.9A CN111233714B (en) | 2020-03-18 | 2020-03-18 | Preparation method of MAPS polypeptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010190425.9A CN111233714B (en) | 2020-03-18 | 2020-03-18 | Preparation method of MAPS polypeptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111233714A CN111233714A (en) | 2020-06-05 |
| CN111233714B true CN111233714B (en) | 2022-04-08 |
Family
ID=70877137
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010190425.9A Active CN111233714B (en) | 2020-03-18 | 2020-03-18 | Preparation method of MAPS polypeptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111233714B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0331073A2 (en) * | 1988-02-29 | 1989-09-06 | Chugai Seiyaku Kabushiki Kaisha | Benzhydrylamine derivatives |
| CN1235163A (en) * | 1998-05-08 | 1999-11-17 | 北京医科大学 | Peptide for vaccine of schistosomiasis |
| CN1599577A (en) * | 2001-07-30 | 2005-03-23 | 埃匹克斯医疗股份有限公司 | Peptide-based multimeric targeted contrast agents |
| WO2006136460A2 (en) * | 2005-06-22 | 2006-12-28 | Glaxo Group Limited | New adjuvant |
| CN101503460A (en) * | 2009-03-12 | 2009-08-12 | 上海交通大学 | IgE antigenicity sequence and preparation of branched multiple antigenic peptides |
| CN106916086A (en) * | 2015-12-28 | 2017-07-04 | 深圳翰宇药业股份有限公司 | A kind of solid phase synthesis process of paranitroanilinum modified polypeptide C-terminal |
| CN108676070A (en) * | 2018-05-11 | 2018-10-19 | 滨海吉尔多肽有限公司 | A kind of solid-phase synthesis according to Nagqu peptide |
| CN110317257A (en) * | 2019-06-03 | 2019-10-11 | 吉尔生化（上海）有限公司 | A kind of solid-liquid phase synthesi of sincalide |
| CN110546156A (en) * | 2017-04-20 | 2019-12-06 | 塞特兰斯有限公司 | Method for removing bacterial toxins from biological fluids |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2219646A4 (en) * | 2007-12-21 | 2010-12-22 | Univ Rochester | Method for altering the lifespan of eukaryotic organisms |
-
2020
- 2020-03-18 CN CN202010190425.9A patent/CN111233714B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0331073A2 (en) * | 1988-02-29 | 1989-09-06 | Chugai Seiyaku Kabushiki Kaisha | Benzhydrylamine derivatives |
| CN1235163A (en) * | 1998-05-08 | 1999-11-17 | 北京医科大学 | Peptide for vaccine of schistosomiasis |
| CN1599577A (en) * | 2001-07-30 | 2005-03-23 | 埃匹克斯医疗股份有限公司 | Peptide-based multimeric targeted contrast agents |
| WO2006136460A2 (en) * | 2005-06-22 | 2006-12-28 | Glaxo Group Limited | New adjuvant |
| CN101503460A (en) * | 2009-03-12 | 2009-08-12 | 上海交通大学 | IgE antigenicity sequence and preparation of branched multiple antigenic peptides |
| CN106916086A (en) * | 2015-12-28 | 2017-07-04 | 深圳翰宇药业股份有限公司 | A kind of solid phase synthesis process of paranitroanilinum modified polypeptide C-terminal |
| CN110546156A (en) * | 2017-04-20 | 2019-12-06 | 塞特兰斯有限公司 | Method for removing bacterial toxins from biological fluids |
| CN108676070A (en) * | 2018-05-11 | 2018-10-19 | 滨海吉尔多肽有限公司 | A kind of solid-phase synthesis according to Nagqu peptide |
| CN110317257A (en) * | 2019-06-03 | 2019-10-11 | 吉尔生化（上海）有限公司 | A kind of solid-liquid phase synthesi of sincalide |
Non-Patent Citations (4)
| Title |
|---|
| Reduced amide bond neurotensin 8-13 mimetics with potent in vivo activity;Wustrow, D. J. et al;《Bioorganic & Medicinal Chemistry Letters》;19921231;第5卷(第9期);997-1002 * |
| Synthesis of tripeptides as potent Yersinia protein tyrosine phosphatase inhibitors;Kyeong Lee et al;《Bioorganic & Medicinal Chemistry Letters》;20051231;第15卷;4037-4042 * |
| trans-Hexahydrobenzoxazolidinones in the Enantioselective Synthesis of β2-Amino Acids Containing Proteinogenic Side Chains;Yamir Bandala et al;《European Journal of Organic Chemistry》;20141231;2275-2283 * |
| α芋螺毒素MI分支肽的合成及免疫原性研究;陈荣芳等;《生物技术通讯》;20191231;第30卷(第6期);768-774 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111233714A (en) | 2020-06-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4405594B2 (en) | Improved solid phase peptide synthesis and reagents for use in such synthesis | |
| CN103374054B (en) | One-step method based solid-phase polypeptide synthesis method | |
| CN110294800B (en) | Preparation method of somaglutide | |
| JP2003530311A (en) | Methods and compositions for peptide synthesis | |
| CN113801197B (en) | Preparation method of procalcitonin | |
| CN107880111A (en) | A kind of method for preparing Liraglutide | |
| CN107056927A (en) | A kind of preparation method of Liraglutide | |
| CN112111002B (en) | Preparation method of semaglutide | |
| CN111087462B (en) | Solid-phase synthesis method of abamectin | |
| CN108218957B (en) | Method for preparing AMG416 by combining solid phase and liquid phase | |
| CN106632655B (en) | Preparation method of exenatide and product thereof | |
| CN112111001B (en) | Method for synthesizing thymosin T alpha-1 | |
| CN106478805A (en) | A kind of preparation method of GLP-1 derivant | |
| CN114981285B (en) | Synthesis method of C-terminal protection fragment of peptide | |
| CN113444150A (en) | Solid-phase preparation method of procatide | |
| CN112585153A (en) | Compound or salt thereof, and preparation method and application thereof | |
| CN111233714B (en) | Preparation method of MAPS polypeptide | |
| CN110642936B (en) | Method for preparing teriparatide | |
| CN108676082A (en) | A kind of solid-phase synthesis of beta-amyloyd peptide 1-42 | |
| CN111944040B (en) | Solid phase synthesis method of abamectin | |
| CN103936848B (en) | Thymosin α1Method of synthesis of | |
| JP2008534639A (en) | Peptide synthesis of alpha-helix on PEG resin | |
| CN112175067B (en) | Preparation method of teduglutide | |
| WO2021007703A1 (en) | A method for preparing liraglutide via a solid phase peptide synthesis | |
| CN111892650A (en) | Solid-phase synthesis method of liraglutide |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |