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A kind of purification method of carbetocin

A carbetocin and purification method technology, applied in the purification field of carbetocin, can solve the problems such as difficulty in separating and obtaining high-purity products, complicated purification and purification process, high cost and the like, and shorten the purification process. route, improve purification efficiency, reduce the number of purification times and the effect of steps

Active Publication Date: 2021-05-28
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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  • A kind of purification method of carbetocin
  • A kind of purification method of carbetocin
  • A kind of purification method of carbetocin

Examples

Experimental program
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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 kind of purification method of carbetocin, comprising the following steps: A) dissolving the crude carbetocin in the mixed solution of acetonitrile-purified water, filtering to obtain the first crude product solution; B) adjusting The pH value of the first crude product solution is 2.0~2.5, and under the condition of 2~8 ℃, precipitates solid, filters to obtain the second crude product solution; C) adjust the pH value of the second crude product solution to 7.5~8.5, at 2~8 Under the condition of ℃, the solid is precipitated, filtered to obtain the third crude product solution; D) the pH value of the third crude product solution is adjusted to 3.5-4.5, and the solid is precipitated under the condition of 2-8 ℃, filtered to obtain the initial carbetocin Pure solution; E) Purify the primary carbetocin solution by reverse phase chromatography to obtain a pure carbetocin solution. The above-mentioned purification method provided by the present invention. According to the purification method provided by the present invention, the product purity can reach more than 99.5%, and the yield can reach more than 69%.

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 Patents(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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