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Oxytocin purifying method

A purification method and oxytocin technology, applied in the field of medicine, can solve the problems of low titer and low yield, and achieve the effects of simple operation, improved product purity, and improved purity

Inactive Publication Date: 2018-05-18
宁波人健药业集团股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the yield of this method is not high, and the titer per milliliter is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] A kind of purification method of oxytocin, the method comprises the following steps:

[0053] 1. Nonapeptide-intermediate production process

[0054] 1.1. Preparation

[0055] Pour about 600ml of liquid ammonia into a 1L three-neck round bottom flask, place the reaction flask on a magnetic stirrer, then add 16g of nonapeptide intermediate, stir to dissolve, and add an appropriate amount of ammonium chloride;

[0056] 1.2. Vacuum drying

[0057] Turn on the vacuum pump and unplug the drying tube on the reaction bottle. After the volatile ammonia gas is exhausted, put the elbow into the mouth of the bottle, and control the degree of vacuum so as not to draw out the liquid. Until the liquid is drained, the white powder on the wall of the reaction bottle falls off.

[0058] 1.3. Dissolving

[0059] Prepare 20L of 2-4% glacial acetic acid solution in a stainless steel bucket, add 800ml into the dried reaction bottle, dissolve the dry powder and clean it, pour the soluti...

Embodiment 2

[0079] A purification method of oxytocin, different from Example 1, the procedure for setting the gradient elution is shown in Table 2:

[0080] Table 2 Gradient elution program

[0081] time / min

[0082] Other steps are the same as in Example 1, and the quality detection results of the prepared oxytocin are shown in Table 4.

Embodiment 3

[0084] A purification method of oxytocin, which is different from Example 1: the procedure for setting the gradient elution is shown in Table 3:

[0085] Table 3 Gradient elution program

[0086] time / min

A％

B％

0

98

2

25

98

2

75

90

10

76

80

20

90

80

20

[0087] Other steps are the same as in Example 1, and the quality detection results of oxytocin prepared in Example 1-Example 3 are shown in Table 4.

[0088] The oxytocin quality detection that table 4 embodiment 1-embodiment 3 prepares

[0089]

[0090] The results in Table 4 prove that, under other conditions being the same, the optimal program for gradient elution is set as shown in Table 1, and the titer of oxytocin is the highest. On this basis, regardless of the increase in mobile phase A Or reduce, the potency of oxytocin obtained is significantly reduced.

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Abstract

The invention relates to an oxytocin purifying method. The oxytocin purifying method comprises the following steps: (1) drying liquid ammonia in vacuum; (2) adding an acid solution into a dried reaction flask, dissolving a dry powder; (3) introducing air or oxygen into the lower part of a liquid obtained in the step (2) to oxidize, obtaining an intermediate of an oxytocin solution; (4), purifyingthe intermediate of the oxytocin solution through high phase liquid chromatographic column; (5) rotatably evaporating an effective component eluant collected in the step (4). The various components inthe oxytocin solution have different distribution coefficients in the mobile phase and the stationary phase, and are separated into single components, the single components sequentially flow out froma column, and the required oxytocin component is collected. An organic solvent in the oxytocin is evaporated through rotary evaporation, and the oxytocin is separated. The preparation method is simple in operation and high in efficiency, the purity of the obtained oxytocin is high, the single impurity is less than or equal to 2.0%, the total impurity is less than or equal to 5%, and the unit titer is greater than 1500 unit / ml.

Description

technical field [0001] The invention belongs to the technical field of medicine and relates to a method for purifying oxytocin. Background technique [0002] Oxytocin solution, also known as oxytocin, is a raw material drug, molecular formula: C 43 h 66 N 12 o 12 S 2 , Molecular weight: 1007.19, is composed of 9 amino acid residues, a biologically active polypeptide amino acid product. Among them, two cysteines form a disulfide bond at positions 1 and 6, with a relative molecular mass of 1007.2 and an isoelectric point of 7.7. It is stable in acidic solutions and exists in the form of free peptides in blood circulation. Soluble in water, soluble in acetone, dilute acetic acid and butanol, slightly soluble in acetonitrile. Oxytocin is mainly used clinically to induce labor, postpartum hemostasis, and intrapartum uterine atony. In addition, oxytocin also has a wide range of physiological functions, especially on the central nervous system. Structural formula: [0003]...

Claims

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Application Information

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IPC IPC(8): C07K7/16C07K1/16C07K1/14
CPCC07K7/16
Inventor 夏星辉王群芳胡晶锦
Owner 宁波人健药业集团股份有限公司
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