Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Solid phase synthesis method for adrenomedullin (27-52)

An adrenomedullin and solid-phase synthesis technology, which is applied in the field of solid-phase synthesis of adrenomedullin, can solve the problems of excessive production of side reaction products, shortening the service life of equipment, unstable side chain protecting groups, etc., and achieves side reactions. Less product production, easy purification, and easy monitoring and control of the effect of the reaction

Inactive Publication Date: 2011-04-20
WEST CHINA HOSPITAL SICHUAN UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 1. During the synthesis process, it is necessary to repeatedly use acid to remove the protecting group, and then use hydrogen fluoride to crack the peptide from the resin, so the reaction conditions are severe, which will inevitably cause side reactions, and make some of the side chain protecting groups ineffective. Stable, resulting in the gradual loss of the extended peptide chain from the solid support, low yield, and changes in the side chains of amino acid residues such as Asp (aspartic acid), Lys (lysine), etc.
[0008] 2. Due to the production of more side reaction products, the difficulty of purification is increased
[0009] 3. Since the hydrogen fluoride used is a strong acid and highly corrosive, it not only shortens the service life of the equipment, but also pollutes the environment

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] This embodiment is the first peptide grafted resin.

[0027]Weigh 0.25mmol (234mg) p-alkoxybenzyl alcohol resin and place it in a solid-phase reactor, soak it with dichloromethane for 2h and filter it with suction, then add 1mmol Fmoc-Ala-OH, 1mmol dicyclohexylcarbodiimide , 0.1mmol 4-dimethylaminopiperidine was dissolved in 6mL of N-methylpyrrolidone, added to a solid-phase reactor and reacted with p-alkoxybenzyl alcohol resin at room temperature for 2h, after the reaction was completed, dichloromethane Wash 5 times with 3ml each time, then add 4ml 20% piperidine / dimethylformamide mixed solution, shake for 0.5h, filter with suction, wash 5 times with 2ml each time of N-methylpyrrolidone, dichloromethane each time Wash 5 times with 2ml each time and wash 5 times with 3ml methanol each time, and repeat the above washing-deprotection-washing twice, and finally the reactant is drained to constant weight.

Embodiment 2

[0029] This example is the synthesis of the second peptide.

[0030] First, add 1-hydroxybenzotriazole, 1-hydroxy-benzo-triazole tetrazole prepared in 1:1:1 according to the molar ratio of the resin to the solid-phase reactor with the resin obtained in the previous step reaction. Methyl hexafluorophosphate, diisopropylethylamine condensation reagent 2mL (concentration 0.45mol / L) and 0.5mmol Fmoc-His(Trt)-OH, and add 6ml N-methylpyrrolidone, condense at room temperature for 2h, Then wash with 3ml of dichloromethane for 5 times, then add 4ml of 20% piperidine / dimethylformamide mixed solution, shake for 0.5h, filter with suction, and wash with 2ml of N-methylpyrrolidone for 5 times times, dichloromethane each 2ml wash 5 times and methanol each 3ml wash 5 times, and repeat the above washing-deprotection-washing twice, and finally the reactant was drained to constant weight.

Embodiment 3～26

[0032] This part of the examples is based on Example 2 to continue the synthesis of the third peptide to the twenty-sixth peptide.

[0033] Since the synthetic procedures and control conditions of these examples are completely the same as those of Example 2 except for the addition of protected amino acids, the description is omitted here. It is worth noting that the added protected amino acids are all 0.5 mmol, and the added protected amino acids are as follows:

[0034] Fmoc-Gln(Trt)-OH;

[0035] Fmoc-Ile-OH;

[0036] Fmoc-Tyr(tBu)-OH;

[0037] Fmoc-Gln(Trt)-OH;

[0038] Fmoc-Phe-OH;

[0039] Fmoc-Thr(tBu)-OH;

[0040] Fmoc-Asp(OtBu)-OH;

[0041] Fmoc-Lys-(Boc)-OH;

[0042] Fmoc-Asp(OtBu)-OH;

[0043] Fmoc-Lys-(Boc)-OH;

[0044] Fmoc-Asp(OtBu)-OH;

[0045] Fmoc-Asn(Trt)-OH;

[0046] Fmoc-Val-OH;

[0047] Fmoc-Ala-OH;

[0048] Fmoc-Pro-OH;

[0049] Fmoc-Arg(Pbf)-OH;

[0050] Fmoc-Ser(tBu)-OH;

[0051] Fmoc-Lys-(Boc)-OH;

[0052] Fmoc-Ile-OH;

[0053] Fmoc-Ser...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention discloses solid phase synthesis process of adrenal medullarin (27-52). The synthesis process includes the first solid phase Fmoc polypeptide synthesis to graft protective amino acid Fmoc-Ala-OH onto resin, connecting the rest protective amino acids successively, cutting down the synthesized coarse peptide with peptide cutting reagent, and final separating and purifying to obtain the adrenal medullarin (27-52) as the target peptide. The present invention has mild reaction condition, high reaction rate, less side reactions, easy purification, high yield, no corrosion to the equipment and no environmental pollution.

Description

technical field [0001] The invention belongs to the technical field of solid-phase synthesis method and application of polypeptide, and in particular relates to a solid-phase synthesis method of adrenomedullin (27-52). Background technique [0002] Adrenomedullin (ADM) is expressed in the normal human adrenal gland, and it is also highly expressed in bone tissue and secreted by osteoblasts. [0003] Adrenomedullin is a potent stimulator of osteoblastic activity invitro and in vivo. Am J Physiol 1997; 273: E1113- 20) Studies have found that it shares 24% homology with calcitonin gene-related peptide (CGRP), a drug for treating osteoporosis, and should belong to the CGRP family; human ADM is Composed of 52 amino acid residues, the 16th and 21st cysteine ​​residues constitute a ring structure of disulfide bonds in the chain. This structural feature is very important for its biological activity, but it is suggested that the ring structure is not Necessary for osteoblast prolif...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/575C07K1/04
CPCY02P20/55
Inventor 张晖陈治宇裴福兴王怡鑫
Owner WEST CHINA HOSPITAL SICHUAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com