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Purification method of carbetocin

A carbetocin and purification method technology, which is applied in the field of carbetocin purification, can solve the problems of difficult separation to obtain high-purity products, complex purification and refining process, and high cost, and shorten the purification process route, improvement of purification efficiency, production cycle and the effect of cost reduction

Active Publication Date: 2020-09-15
CHENGDU SINTANOVO BIOTECHNOLOGV CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Commonly used purification methods for carbetocin include reversed-phase high-performance liquid chromatography, gel method, ion exchange method, etc., and the above methods are difficult to separate and obtain high-purity products (>99.5%), or the yield is relatively low (< 50%), or the purification and refining process is complex and the cost remains high

Method used

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  • Purification method of carbetocin
  • Purification method of carbetocin
  • Purification method of carbetocin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] (1) Sample treatment: The crude carbetocin peptide with a chromatographic purity of 60% to 70% was prepared by solid-phase synthesis, and 3L of a mixed solution of purified water-acetonitrile (1:1) was used, according to 15g to 20g / L The concentration is dissolved, stirred, and after complete dissolution, filter with a 0.45um water-based filter membrane to remove insoluble matter and obtain the filtrate, which is the crude carbetocin solution.

[0053] The HPLC spectrogram of the obtained carbetocin crude product solution is as follows figure 1 shown.

[0054] (2) Salt precipitation:

[0055] First, add trifluoroacetic acid solution (1.0%) to adjust the pH value of the crude product solution to 2.0-2.5, and place it at 2-8°C for refrigerated storage for 2-4h, filter off the gray precipitate; then adjust the pH value of the crude product solution with alkali (ammonia water) solution The value is 7.5-8.5, and stored at 2-8°C for 2-4 hours, filtered to remove the precipi...

Embodiment 2

[0065] (1) Sample processing:

[0066] 55g of carbetocin crude peptide with a chromatographic purity of 60% was obtained by solid-phase synthesis, dissolved in 3L of purified water-acetonitrile (1:1) mixed solution according to the concentration of 15g-20g / L, stirred, and completely dissolved Afterwards, filter with a 0.45um water-based filter membrane to remove insoluble matter and obtain the filtrate, which is the crude carbetocin solution.

[0067] (2) Salt precipitation:

[0068] First add TFA solution (0.5%) to adjust the pH value of the crude product solution to 2.0-2.5, and place it at 2-8°C for refrigerated storage for 2-4h, filter off the gray precipitate; then use alkali (ammonia) solution to adjust the pH value of the crude product solution. 7.5-8.5, and stored at 2-8°C for 2-4 hours, filtered to remove the precipitate; then use TFA solution (0.5%) to adjust the pH value of the crude product solution to 3.5-4.5, and store at 2-8°C for 2 Filter after -4h.

[0069]...

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Abstract

The invention provides a purification method of carbetocin. The purification method comprises the following steps: A) dissolving a carbetocin crude product in an acetonitrile-purified water mixed solution, and performing filtering to obtain a first crude product solution; B) adjusting the pH value of the first crude product solution to 2.0-2.5, separating out a solid at 2-8 DEG C, and performing filtering to obtain a second crude product solution; C) adjusting the pH value of the second crude product solution to 7.5-8.5, separating out the solid at 2-8 DEG C, and performing filtering to obtaina third crude product solution; D) adjusting the pH value of the third crude product solution to 3.5-4.5, separating out the solid at 2-8 DEG C, and performing filtering to obtain a crudely purifiedsolution of carbetocin; and E) performing fine purification on the crudely purified solution of the carbetocin through a reversed phase chromatography to obtain a finely purified solution of the carbetocin. According to the purification method provided by the invention, the product purity reaches 99.5% or above, and meanwhile, the yield can reach 69% or above.

Description

technical field [0001] The invention relates to the technical field of compound purification, in particular to a method for purifying carbetocin. Background technique [0002] Carbetocin, a synthetic long-acting nonapeptide analogue of oxytocin with agonist properties, can be clinically administered as a single intravenous dose immediately after cesarean section under epidural or spinal anesthesia to prevent uterine hypotonia and postpartum hemorrhage. [0003] The clinical and pharmacological properties of carbetocin are similar to those of naturally occurring oxytocin. Like oxytocin, carbetocin binds to the oxytocin receptor of uterine smooth muscle, causing rhythmic contraction of the uterus, increasing its frequency and increasing uterine tension on the basis of the original contraction. Oxytocin receptor levels in the uterus are low in the non-pregnant state, increase during pregnancy, and peak at parturition. Carbetocin therefore has no effect on the non-pregnant ut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/16C07K1/36C07K1/34C07K1/30C07K1/20
CPCC07K7/16
Inventor 左成罗磊赵建波王彬容付晓平钟国庆黄波高剑李元波袁瑜
Owner CHENGDU SINTANOVO BIOTECHNOLOGV CO LTD
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