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Method for preparing angiotensin and angiotensin acetate

An angiotensin and desalination technology, applied in the field of biopharmaceuticals, can solve the problems of complex preparation process and low yield, and achieve the effect of optimizing the production process

Active Publication Date: 2014-08-06
SPH NO 1 BIOCHEM & PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a preparation method of polypeptide and polypeptide salt in order to overcome the defects of complex preparation process and low yield of polypeptide salt in the prior art

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1 (HPLC method detects angiotensin crude product raw material and purification intermediate solution purity and quantification)

[0036] Instrument: Waters2695 / 2489 high performance liquid chromatography

[0037] Separation column: Waters XBridge-C18, 4.6×150mm, 5μm

[0038] Mobile phase: A is 20mM NaH 2 PO 4 -H 3 PO 4(pH3.0), B is 50% aqueous acetonitrile solution by volume percentage

[0039] Flow rate is 1.0mL / min, detection wavelength is 220nm, room temperature detection,

[0040] The elution gradient is shown in Table 1 below, and the percentages are volume percentages.

[0041] Table 1 Mobile phase elution gradient

[0042] Elution step

Embodiment 2

[0043] Embodiment 2 (75mm inner diameter L&L4003 preparative column packing)

[0044] Using Load&Lock dynamic axial compression and static locking technology, the filler is styrene-divinylbenzene copolymer (reversed phase packing Agilent PLRP-S), the pore size is 10nm, the particle size is 10μm, and the column packing density is 0.33g / mL. Bed pressure 650psi, using Varian chromatographic packing system, 370g dry powder filler, 2L methanol after stirring and homogenizing, poured into 75mm inner diameter L&L4003 preparative column, compression ratio is 3:1, carrier gas is N 2 , adjust the carrier gas pressure so that the oil pressure gauge pressure is 2000psi, and the dynamic axial compression to the height of the column bed is 26cm, which is used as the preparative column for the reverse-phase purification and reverse-phase desalination scheme.

Embodiment 3

[0046] (1) Reversed-phase purification of crude angiotensin raw materials

[0047] Instrument: Varian SD-1 high pressure liquid phase preparation system

[0048] Chromatographic column: self-packed preparative column Load&Lock400375×260mm in Example 2, PLRP-S10μm10nm

[0049] Mobile phase: A is a 0.1% trifluoroacetic acid aqueous solution by volume, B is a 0.1% trifluoroacetic acetonitrile solution by volume, angiotensin crude product (angiotensin synthesized by solid-phase method disclosed in patent application CN201210131066.5) Crude product) through the acetonitrile aqueous solution of volume percent 5% is that solvent is dissolved and is mixed with 10g / L solution, sample liquid is mobile phase C after filtration clarification, measures its HPLC purity by the method for Example 1 and is 84.98%, and retention time is 10.34min , the HPLC data are shown in Table 2.

[0050] Table 2 The peak results of angiotensin crude product HPLC detection

[0051] Peak ID

re...

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Abstract

The invention discloses a method for preparing angiotensin and angiotensin acetate. The method for preparing angiotensin comprises the steps of carrying out reverse phase purification and reverse phase desalination on a crude angiotensin solution sequentially by virtue of a high-performance-liquid reverse-phase chromatography, wherein filler used in the high-performance-liquid reverse-phase chromatography is polystyrene-divinyl benzene (PS-DVB) copolymer. According to the method, in combination with reverse phase purification and reverse phase desalination, a novel application of polystyrene-divinyl benzene as polymer filler is designed, and the angiotensin and the angiotensin acetate can be prepared on a large scale.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals. More specifically, the present invention relates to the preparation method of angiotensin and angiotensin acetate. Background technique [0002] Angiotensin (or angiotensin amine) is a blood pressure-increasing drug, the English name angiotensinamide or [Asn 1 , Val 5 ]-angiotensinⅡ, chemical structure L-Asn-L-Arg-L-Val-L-Tyr-L-Val-L-His-L-Pro-L-Phe, is a synthetic polypeptide composed of eight amino acid residues , Theoretical molecular weight is 1031.18. Angiotensin is used for trauma or postoperative shock and hypotension caused by general anesthesia or lumbar anesthesia, etc., can prevent or treat perioperative hypotension, and can be used as emergency medicine to treat shock hypotension, and can also be used to treat vascular tension In case of overdose of the conversion enzyme inhibitor and conventional treatment is ineffective. [0003] Vasopressin tannin is a synthetic polypeptide c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/14C07K7/16C07K7/06C07K1/20
Inventor 洪勇江锡铭丁金国黄臻辉
Owner SPH NO 1 BIOCHEM & PHARMA CO LTD
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