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Purifying method for carbetocin

A carbetocin and purification method technology, which is applied in the purification field of polypeptide drugs, can solve the problems of low yield, low purity, difficult purification and the like, and achieve the effects of simplifying purification steps, simple operation and reducing cost.

Inactive Publication Date: 2016-03-16
郑州大明药物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] In order to overcome the technical problems of low yield, low purity and difficult purification in the prior art, the inventor has carried out a large amount of research, thus completing the present invention

Method used

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  • Purifying method for carbetocin
  • Purifying method for carbetocin
  • Purifying method for carbetocin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Step 1. Take 5g of carbetocin crude peptide and add 50ml of 1% acetic acid solution to prepare a 10% carbetocin crude peptide solution.

[0036] Step 2, the acetic acid solution of carbetocin is loaded into the 500 Dalton RC semi-permeable membrane with a molecular weight cut-off, and the loading is 70% of the volume of the semi-permeable membrane, and placed in 2L acetate buffer ( pH3.5) buffer solution, stirred and diafiltered at 20~30℃ until the buffer solution pH=2.0~3.0.

[0037] Step 3. Transfer the semi-permeable membrane of the acetic acid solution containing carbetocin to 3L of purified water, stir and diafilter at 20-30°C until the solution pH in the semi-permeable membrane bag is 3.5-4.5.

[0038] Step 4, the carbetocin solution in the semipermeable membrane is taken out, purified by reverse phase chromatography, and the elution fraction containing carbetocin is collected, and the specific chromatographic conditions are as follows;

[0039] Chromatographic c...

Embodiment 2

[0047] Step 1. Take 5g of carbetocin crude peptide and add 50ml of 1% acetic acid solution to prepare a 10% carbetocin crude peptide solution.

[0048] Step 2, the acetic acid solution of carbetocin is loaded into the CE semipermeable membrane with a molecular weight cut-off of 500 Daltons, and the loading is 70% of the volume of the semipermeable membrane, and placed in 2L acetate buffer ( pH3.5) buffer solution, stirred and diafiltered at 20~30℃ until the buffer solution pH=2.0~3.0.

[0049] Step 3. Transfer the semi-permeable membrane of the acetic acid solution containing carbetocin to 3L of purified water, stir and diafilter at 20-30°C until the solution pH in the semi-permeable membrane bag is 3.5-4.5.

[0050] Step 4, the carbetocin solution in the semipermeable membrane is taken out, purified by reverse phase chromatography, and the elution fraction containing carbetocin is collected, and the specific chromatographic conditions are as follows;

[0051] Chromatographic...

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Abstract

The invention discloses a purifying method for carbetocin. The method employs a purifying scheme of a combination of semi-permeable membrane dialysis purification and reversed phase chromatographic purification to eventually obtain carbetocin with a purity of greater than 99.5%. Semi-permeable membrane dialysis purification is simple to operate and does not need special equipment, and a semi-permeable membrane is repeatedly usable, so cost is reduced. Meanwhile, through preliminary purification by using semi-permeable membrane technology, chromatographic purification can be carried out without concentration, so the step of high-temperature concentration is omitted, the possibility of high-temperature degradation of polypeptide is reduced, and yield and purity are ensured.

Description

technical field [0001] The invention relates to a purification method of polypeptide drugs, in particular to a purification method for synthesizing carbetocin. Background technique [0002] Carbetocin is a cyclic polypeptide with the following structural formula. [0003] [0004] The chemical structure is simplified as follows. [0005] [0006] Carbetocin is a synthetic long-acting nonapeptide analogue of oxytocin with agonist properties. It has no effect on the non-pregnant uterus, but has potent uterotonic effects on the pregnant uterus and the newly delivered uterus. Clinical and pharmacological properties are similar to those of naturally occurring oxytocin. Like oxytocin, carbetocin binds to the oxytocin receptors of uterine smooth muscle, causing rhythmic contractions of the uterus, increasing its frequency and increasing uterine tension on the basis of the original contractions. Oxytocin receptor levels in the uterus are low in the non-pregnant state, incre...

Claims

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Application Information

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IPC IPC(8): C07K7/16C07K1/36C07K1/34C07K1/20
CPCC07K7/16
Inventor 毛影李运铎李沁沁朱赞梅张佳美娄丽丽
Owner 郑州大明药物科技有限公司
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