Method for purifying Exenatide

A technology of exenatide and crude peptide, which is applied in the field of HPLC, can solve the problems of exenatide research and achieve high yield and high purity

Inactive Publication Date: 2009-09-23
HYBIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are few related literatures on the solid-phase direct synthesis of Exendin-4 at home and abroad, but there are no literatures that specifically study the methods of Exenatide-related purification, especially the method of large-scale purification of Exenatide

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1. Sample treatment: Dissolve the synthetic exenatide crude peptide in water for injection (concentration is about 50 mg / mL, filter through a filter membrane with a pore size of 0.45 μm, and collect the filtrate for later use.

[0021] 2. Purification:

[0022] Purification conditions: chromatographic column: a chromatographic column with octaalkylsilane bonded silica gel as the stationary phase, and the diameter and length of the column are: 5cm×25cm. Mobile phase: Phase A: aqueous solution of sodium dihydrogen phosphate, adjust the pH to 3.5-4.3 with analytically pure phosphoric acid solution; phase B: acetonitrile of chromatographically pure. Flow rate: 50-70ml / min. Detection wavelength: 230nm. Gradient: B%: 20% ~ 48% 66-78min. The injection volume is 1.3-2g.

[0023] Purification process: the chromatographic column is rinsed with more than 50% acetonitrile and then loaded with a sample volume of 26-40ml of sample solution. Linear gradient elution, collect the t...

Embodiment 2

[0026] 1. Sample treatment: Dissolve the synthetic exenatide crude peptide in water for injection (concentration is about 50 mg / mL, filter through a filter membrane with a pore size of 0.45 μm, and collect the filtrate for later use.

[0027] 2. Purification:

[0028] Purification conditions: chromatographic column: a chromatographic column with tetraalkylsilane bonded silica gel as the stationary phase, and the diameter and length of the column are: 5cm×25cm. Mobile phase: Phase A: aqueous solution of sodium dihydrogen phosphate, adjust the pH to 3.0-4.8 with analytically pure phosphoric acid solution; phase B: acetonitrile of chromatographically pure. Flow rate: 50-70ml / min. Detection wavelength: 230nm. Gradient: B%: 10% ~ 35% 50-75min. The injection volume is 1.1-1.8g.

[0029] Purification process: the chromatographic column is rinsed with more than 50% acetonitrile and then loaded with a sample volume of 26-40ml of sample solution. Linear gradient elution, collect th...

Embodiment 3

[0032] 1. Sample treatment: Dissolve the synthetic exenatide crude peptide in water for injection (concentration is about 50 mg / mL, filter through a filter membrane with a pore size of 0.45 μm, and collect the filtrate for later use.

[0033] 2. Purification:

[0034] 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: aqueous solution of sodium dihydrogen phosphate, adjust the pH to 3.8-5.3 with analytically pure phosphoric acid solution; phase B: acetonitrile of chromatographically pure. Flow rate: 50-70ml / min. Detection wavelength: 230nm. Gradient: B%: 15% ~ 55% 60-78min. The injection volume is 1.5-2.0g.

[0035] Purification process: the chromatographic column is rinsed with more than 50% acetonitrile and then loaded with a sample volume of 30-35ml sample solution. Linear gradient elution, collect the ta...

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PUM

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Abstract

The invention discloses a method for purifying Exenatide, which comprises the following steps of: 1) dissolving crude peptides obtained by solid phase synthesis with water for injection; 2) conducting gradient elution and purification with the fixed phase being reversed-phase silica gel column of tetraalkylsilane bonded silica, octalkylsilane bonded silica or octadecylsilane bonded silica, and the phase A of a mobile phase being phosphate buffer solution and the phase B of the mobile phase being chromatographic grade acetonitrile, and collecting the peptide solution at a target peak value; and 3) converting the high-purity peptides after purification into acetate by using an anion exchange method. The method which is applicable to the industrialized purification of Exenatide uses reversed phase high-performance liquid chromatography for purifying Exenatide, and uses the anion exchange method for converting the high-purity peptides after purification into acetate, thus not only being capable of obtaining the refined peptides with PLC purity higher than 98.0 percent, but also realizing large-scale production and meeting the requirements of high purity, high yield and industrialization.

Description

technical field [0001] The invention belongs to the technical field of HPLC, in particular to a method for large-scale purification of Exenatide. Background technique [0002] Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia caused by a variety of etiologies. Hyperglycemia is mainly caused by defects in insulin secretion or action. Diabetes can be divided into two types, insulin-dependent diabetes (type I diabetes) and non-insulin-dependent diabetes (type II diabetes), wherein type II diabetes patients account for more than 90%. According to the statistics of WHO, there are currently 130 million people with type 2 diabetes diagnosed in the world, and my country has more than 40 million people. It is the second largest country with diabetes after India. [0003] Commonly used drugs for treating type II diabetes include: biguanides, sulfonylureas, α-glucosidase inhibitors, thiazolidinediones and insulin. However, studies have found that these ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K1/20A61P3/10
Inventor 康旭覃亮政李红玲马亚平袁建成
Owner HYBIO PHARMA
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