Method for purifying Carbetocin

A carbetocin and solution technology, applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry and other directions

Inactive Publication Date: 2009-09-16
HYBIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there is no specific literature on the method of carbetocin-related purification, especially the large-scale purification method. The present invention can not o...

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Sample treatment: the crude carbetocin (Carbetocin) peptide obtained by solid-phase synthesis was dissolved into 500mg / mL with water for injection, and used After membrane filtration, the filtrate was collected for later use.

[0026] 2. Purification:

[0027] 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 to adjust the pH to 2.8-3.3 with analytically pure phosphoric acid; phase B: acetonitrile of chromatographically pure. Flow rate: 60-80ml / min. Detection wavelength: 230nm. Gradient: B%: 24% ~ 38% 56-64min. The injection volume is 1.5-2g.

[0028] Purification process: the chromatographic column is rinsed with more than 50% acetonitrile and then loaded with a sample volume of 30-40ml of sample solution. After linear gradient elution for 56-64 m...

Embodiment 2

[0032] 1. Sample treatment: Dissolve 500 mg / mL of crude peptide of carbetocin (Carbetocin) obtained by phase synthesis in water for injection, and use After membrane filtration, the filtrate was collected 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: 15cm×25cm. Mobile phase: Phase A: aqueous solution of sodium dihydrogen phosphate, adjust the pH to 2.8-3.3 with analytically pure phosphoric acid solution; phase B: acetonitrile of chromatographically pure. Flow rate: 450-550ml / min. Detection wavelength: 230nm. Gradient: B%: 24% ~ 38% 56-64min. The injection volume is 10-15g.

[0035] Purification process: wash the chromatographic column with more than 50% acetonitrile and load the sample, the sample volume is 200ml-300ml sample solution. After linear gradient elution for 56-64 minutes, ...

Embodiment 3

[0039] 1. Sample treatment: Dissolve 500 mg / mL of crude peptide of carbetocin (Carbetocin) obtained by phase synthesis in water for injection, and use After membrane filtration, the filtrate was collected for later use.

[0040] 2. Purification:

[0041] 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: aqueous solution of sodium dihydrogen phosphate and adjust the pH to 2.8-3.3 with analytically pure phosphoric acid solution; phase B: acetonitrile of chromatographically pure. Flow rate: 20000-2200ml / min. Detection wavelength: 230nm. Gradient: B%: 24% ~ 38% 56-64min. The injection volume is 50-65g.

[0042]Purification process: wash the chromatographic column with more than 50% acetonitrile and load the sample, the sample volume is 900ml-1300ml sample solution. After linear gradient elution for 60-80 min...

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Abstract

The invention provides a technological method suitable for the industrialized purification of Carbetocin, a reversed-phase efficient liquid chromatography is used to purify the Carbetocin to result in high purity and good yield in order to meet industrialized demands. A crude peptide solution of the Carbetocin is filled with materials by a reversed-phase chromatographic column to be a stationary phase, a phosphate buffer solution is regarded as phase A and acetonitrile is regarded as phase B to implement gradient elution purification, wherein, the gradient: B%:20-40%, the pH of the phase A is 2.5-3.5; an anion exchange method is employed to convert the phosphate and trifluoroacetate to acetate. The invention employs one-step reversed-phase efficient liquid chromatography to purify followed by the acetate conversion by an anion exchange column in one step, thus obtaining high-purity acetate Carbetocin at high yield and offering an efficient purification technology for the massive purification and preparation of the peptide raw drugs.

Description

technical field [0001] The invention relates to a method for purifying polypeptides, in particular to a method for purifying carbetocin. Background technique [0002] Carbetocin is a synthetic long-acting nonapeptide analogue of oxytocin with agonist properties. A single dose intravenously may be administered 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 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, increase during pregnancy, and peak at parturition. Carbetocin therefore has no effect on the non-pregnant uterus, but has po...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K1/20C07K1/18A61P15/04
Inventor 程生强覃亮政马亚平袁建成
Owner HYBIO PHARMA
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