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A method for purifying terlipressin

A terlipressin and antioxidant technology, which is applied in the field of polypeptide purification, can solve the problems of low purity and yield of terlipressin, cannot meet industrial production, restrict the application of terlipressin, etc., and achieves simple operation. Easy, high yield, high purity effect

Active Publication Date: 2014-10-15
HYBIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, the purity and yield of terlipressin prepared at present are relatively low, which cannot meet the needs of industrial production, especially the large-scale purification of terlipressin, which restricts the application of terlipressin

Method used

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  • A method for purifying terlipressin
  • A method for purifying terlipressin
  • A method for purifying terlipressin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Terlipressin was obtained by liquid phase synthesis.

[0050] Sample treatment: Dissolve the crude peptide with 25% acetonitrile aqueous solution at a concentration of 100g / L, stir to dissolve the sample completely, filter it with a filter membrane, and collect the filtrate. Add the ascorbic acid reagent according to the ratio of 5% of the weight of the crude peptide, and dilute the volume ratio of the acetonitrile in the crude peptide solution to 10% with water for later use.

[0051] purification:

[0052] Purification conditions: chromatographic column: a chromatographic column with octadecylsilane bonded silica gel as the stationary phase, and the diameter and length of the column are: 5cm×25cm. Mobile phase: Phase A: 0.4% perchloric acid and 0.04% sulfuric acid aqueous solution (v / v), adjust the pH value to 2.8 with 50mM aqueous sodium hydroxide solution; Phase B: acetonitrile, flow rate: 80ml / min, gradient: B%: 10%~40%, detection wavelength: 280nm. The injectio...

Embodiment 2

[0056] Terlipressin was obtained by solid-phase synthesis.

[0057] Sample treatment: Dissolve the crude peptide with 25% methanol aqueous solution at a concentration of 100g / L, stir to dissolve the sample completely, filter it with a filter membrane, and collect the filtrate. Add ascorbic acid reagent at a ratio of 5% of the weight of the crude peptide, and dilute the volume ratio of methanol in the crude peptide solution to 10% with water for use.

[0058] purification:

[0059] Purification conditions: chromatographic column: a chromatographic column with octadecylsilane bonded silica gel as the stationary phase, and the diameter and length of the column are: 15cm×25cm. Mobile phase: A phase: 0.4% perchloric acid and 0.04% sulfuric acid aqueous solution (v / v), adjust the pH value to 2.8 with 50mM sodium hydroxide aqueous solution; B phase: acetonitrile, flow rate: 500ml / min, gradient: B%: 10%~40%, detection wavelength: 280nm. The injection volume is 15-30g.

[0060] Pur...

Embodiment 3

[0063] Terlipressin was obtained by liquid phase synthesis.

[0064] Sample treatment: Dissolve the crude peptide with 25% methanol aqueous solution at a concentration of 100g / L, stir to dissolve the sample completely, filter it with a filter membrane, and collect the filtrate. Add ascorbic acid reagent at a ratio of 5% of the weight of the crude peptide, and dilute the volume ratio of methanol in the crude peptide solution to 10% with water for use.

[0065] purification:

[0066] Purification conditions: chromatographic column: a chromatographic column with octadecylsilane bonded silica gel as the stationary phase, and the diameter and length of the column are: 30cm×25cm. Mobile phase: Phase A: 0.4% perchloric acid and 0.04% sulfuric acid aqueous solution (v / v), adjust the pH value to 2.8 with 50mM aqueous sodium hydroxide solution; Phase B: acetonitrile, flow rate: 3000ml / min, gradient: B%: 10%~40%, detection wavelength: 280nm. The injection volume is 120g.

[0067] Pur...

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Abstract

The present invention relates to the field of polypeptide purification, and particularly to a method for purifying terli. According to the method, a terli crude product is pre-treated, and then purification and salt conversion are performed to obtain the terli with purity more than 99.5%. The method of the present invention has the following advantages that: the operation is simple and easy to perform; the terli prepared by the method of the present invention has characteristics of high purity (the purity can be more than 99.5%, and the maximum single impurity is less than 0.1%) and high yield (the purification yield can be more than 85%, and the total yield can be more than 50%); and the method meets requirements of industrialization, and more than 1000 g of the refined peptide can be obtained in one batch.

Description

technical field [0001] The invention relates to the field of polypeptide purification, in particular to a method for purifying terlipressin. Background technique [0002] Liver cirrhosis (hepatic cirrhosis) is a common clinical chronic progressive liver disease, which is diffuse liver damage caused by long-term or repeated effects of one or more etiologies. In my country, most of them are post-hepatitis cirrhosis, and a small part are alcoholic cirrhosis and schistosome cirrhosis. Histopathologically, extensive liver cell necrosis, nodular regeneration of residual liver cells, connective tissue hyperplasia, and fibrous septum formation lead to structural destruction of hepatic lobules and formation of pseudolobules. The liver gradually deforms and hardens and develops into cirrhosis. In the early stage, there are no obvious symptoms due to the strong compensatory function of the liver. In the later stage, liver function damage and portal hypertension are the main manifestat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/16C07K1/36C07K1/18C07K1/16
Inventor 赵忠卫覃亮政马亚平袁建成
Owner HYBIO PHARMA
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