Method for preparing ularitide

A technology of ularitide and linear peptide, which is applied in the field of preparation of ularitide, can solve the problems of high cost and inability to increase the yield of ularitide, and achieve the effects of low cost, favorable promotion, and simple method and process

Active Publication Date: 2017-03-22
HANGZHOU GOTOP BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the above method does not improve the synthesis process of ularitide, so the yield of ularitide synthesis cannot be improved, and the finished product with the same purity is obtained, and the cost is higher

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A method for preparing ularitide, the amino acid sequence of said ularitide is shown in SEQ ID No.1: TAPRSLRRSSCFGGRMDRIGAQSGLGCNSFRY;

[0037] The preparation method comprises the following steps:

[0038] 1) Synthesis of fully protected linear peptide resin:

[0039] Using Fmoc-Tyr(tbu)-Wang Resin resin as a solid phase carrier, the Fmoc-Tyr(tbu)-WangResin resin was mixed with DCM, agitated for 30min to fully swell the resin, then the solution was drawn off, and the Fmoc-Tyr(tbu)-WangResin resin was mixed with DCM. Tyr(tbu)-Wang Resin resin was washed three times.

[0040]To Fmoc-Tyr (tbu)-Wang Resin resin, add its volume concentration of 2.5 times volume to the DBLK solution of 20% to react for 5min, then suction filter, wash with DMF, suction filter, add the volume of resin 2.5 times volume again React with 20% DBLK solution for 10 minutes to obtain H-Tyr(tbu)-Wang Resin; wash six times with DMF and remove the solution; take Fmoc-Arg(pdf)-OH and H-Tyr-(tbu)-Wang R...

Embodiment 2

[0052] A method for preparing ularitide, the amino acid sequence of said ularitide is shown in SEQ ID No.1: TAPRSLRRSSCFGGRMDRIGAQSGLGCNSFRY;

[0053] The preparation method comprises the following steps:

[0054] 1) Synthesis of fully protected linear peptide resin:

[0055] Using Wang Resin resin as a solid phase carrier, Fmoc-Tyr(tbu)-OH was coupled on Wang Resin resin to obtain Fmoc-Tyr(tbu)-Wang Resin resin. With Fmoc-Tyr (tbu)-Wang Resin resin as a solid phase carrier, the Fmoc-Tyr (tbu)-Wang Resin resin was mixed and shaken with DMF for 25min to fully swell the resin, then the solution was drawn off, and the Fmoc- Tyr(tbu)-Wang Resin resin was washed three times.

[0056] To the Fmoc-Tyr(tbu)-Wang Resin resin, add its volume concentration of 2 times the volume of the 20% DBLK solution to react for 6min, then filter with suction, wash with DMF, filter with suction, and then add the quality of 2 times the volume of the resin React with 20% DBLK solution for 12 minutes ...

Embodiment 3

[0068] A method for preparing ularitide, the amino acid sequence of said ularitide is shown in SEQ ID No.1: TAPRSLRRSSCFGGRMDRIGAQSGLGCNSFRY;

[0069] The preparation method comprises the following steps:

[0070] 1) Synthesis of fully protected linear peptide resin:

[0071] Using Fmoc-Tyr(tbu)-Wang Resin resin as a solid phase carrier, the Fmoc-Tyr(tbu)-WangResin resin was mixed with DCM, stirred for 35min to fully swell the resin, then the solution was drawn off, and the Fmoc-Tyr(tbu)-WangResin was mixed with DMF Tyr(tbu)-Wang Resin resin was washed three times.

[0072] To Fmoc-Tyr(tbu)-Wang Resin resin, add 3 times its volume concentration to the DBLK solution of 20% to react for 4min, then filter with suction, wash with DMF, filter with suction, then add 3 times of volume of resin React with 20% DBLK solution for 8 minutes to obtain H-Tyr(tbu)-Wang Resin; wash three times with DMF and remove the solution; take Fmoc-Arg(pdf)-OH and H-Tyr-(tbu)-Wang Resin Coupling is ca...

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PUM

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Abstract

The invention relates to the technical field of polypeptide synthesis, and discloses a method for preparing ularitide. The method includes the following steps that (1) full-protection linear peptide resin is synthesized; (2) splitting is performed to obtain linear peptide; (3) oxidation is conducted to obtain the ularitide; and (4) purification, salt rotation and freeze-drying are carried out to obtain the ularitide competitive product. By the adoption of the method, the process is simple, synthesis difficulty of the ularitide can be lowered, the yield of the synthesized ularitide is high, cost is low, the purity is 98% or above, and popularization is facilitated.

Description

technical field [0001] The invention relates to the technical field of polypeptide synthesis, in particular to a preparation method of ularitide. Background technique [0002] Ularitide is a natriuretic cyclic peptide composed of 32 amino acid residues, which was originally isolated from urine by Schulz-Knappe et al. in 1988 and belongs to the atrial natriuretic peptide (ANP) family. Renal natriuretic peptide. Endogenous uraritide is synthesized in the distal renal tubular cells, secreted after the lumen, and binds to the downstream natriuretic peptide A receptor in the inner medullary collecting duct, which can regulate the excretion of sodium and water in the kidney. Therefore, uraritide It has vasodilation and natriuretic effects, and it has been confirmed that ularitide can reduce the reabsorption of urine by the kidneys. The clinical treatment of decompensated heart failure (DHF) is aimed at relieving symptoms and stabilizing the patient's hemodynamics. The currently ...

Claims

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Application Information

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IPC IPC(8): C07K14/58C07K1/20C07K1/06C07K1/04
Inventor 付坤程益明谢振亮沈永良沈永刚
Owner HANGZHOU GOTOP BIOTECH
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