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A kind of refining method of oxytocin [4-glu, 5-asp] impurity

A refining method and 5-asp technology, applied in the preparation method of peptides, chemical instruments and methods, oxytocin/vasopressin, etc., can solve the problem of large amount of hazardous waste liquid, high cost of waste liquid treatment, and uneconomical And other issues

Active Publication Date: 2021-04-13
SPH NO 1 BIOCHEM & PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] The technical problem to be solved by the present invention is to overcome the large amount of hazardous waste liquid produced in the process of refining oxytocin [4-Glu, 5-Asp] impurities in the prior art, all of which are organic hazardous waste liquids, resulting in waste liquid treatment High cost and uneconomical defects, but a purification method for oxytocin [4-Glu, 5-Asp] impurities is provided

Method used

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  • A kind of refining method of oxytocin [4-glu, 5-asp] impurity
  • A kind of refining method of oxytocin [4-glu, 5-asp] impurity
  • A kind of refining method of oxytocin [4-glu, 5-asp] impurity

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Effect test

Embodiment 1

[0055] Embodiment 1 inner diameter 50mm L&L4002 preparative column packing:

[0056] Using Load&Lock dynamic axial compression and static locking technology, the packing is ODS-AQ, pore size 10nm, particle size 10μm, packed to column bed pressure 1000psi, using Varian chromatography packing system, 300g dry powder filler, 600ml isopropanol after stirring and homogenizing, poured into L&L4002 preparative column with inner diameter 50mm, compression ratio 1.5: 1, the carrier gas is N 2 , adjust the carrier gas pressure so that the oil pressure gauge pressure is 1500psi, and the dynamic axial compression to the height of the column bed is 25cm, which is used as a preparative column for reverse-phase enrichment, reverse-phase salt conversion and reverse-phase purification schemes.

Embodiment 2

[0057] Example 2 Reverse-phase enrichment, reverse-phase salt conversion and reverse-phase purification of oxytocin [4-Glu, 5-Asp] impurity crude product solution

[0058] Instrument: Varian SD-1 high pressure liquid phase preparation system

[0059] Chromatographic column: self-packed preparative column Load&Lock400250 × 250mm of embodiment 1, ODS-AQ 10μm10nm

[0060] The structural formula of the oxytocin [4-Glu, 5-Asp] impurity is The concentration of the reduced oxytocin [4-Glu, 5-Asp] impurity crude product solution in the reduced oxytocin [4-Glu, 5-Asp] impurity crude product solution is 1.0mg / ml, and the oxytocin [4-Glu, 5-Asp] The solvent in the impurity crude product solution is an aqueous solution containing trifluoroacetic acid and acetic acid.

[0061] Mobile phase A is 0.02% acetic acid / water by volume, mobile phase B is 0.02% acetic acid / acetonitrile by volume, and sample C1 is the crude product solution of oxytocin [4-Glu,5-Asp] impurity, determined accordi...

Embodiment 3

[0067] Example 3 Reverse Phase Enrichment, Reverse Phase Salt Conversion and Reverse Phase Purification of Oxytocin [4-Glu, 5-Asp] Impurity Crude Solution

[0068] Instrument: Varian SD-1 high pressure liquid phase preparation system

[0069] Chromatographic column: self-packed preparative column Load&Lock4002 50 × 250mm of embodiment 1, ODS-AQ 10μm 10nm

[0070] The structural formula of the oxytocin [4-Glu, 5-Asp] impurity is The concentration of the reduced oxytocin [4-Glu, 5-Asp] impurity crude product solution in the reduced oxytocin [4-Glu, 5-Asp] impurity crude product solution is 1.5mg / ml, and the oxytocin [4-Glu, 5-Asp] The solvent in the impurity crude product solution is an aqueous solution containing trifluoroacetic acid and acetic acid.

[0071] Mobile phase A is 0.05% acetic acid / water by volume, mobile phase B is 0.05% acetic acid / acetonitrile by volume, sample C1 is the crude product solution of oxytocin [4-Glu, 5-Asp] impurity, and the oxytocin [4-Glu, 5-...

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Abstract

The invention discloses a method for refining the impurities of oxytocin [4-Glu, 5-Asp]. Asp] impurity crude product solution is followed by reverse-phase enrichment, reverse-phase salt transfer, and reverse-phase purification; oxytocin [4-Glu,5-Asp] impurity crude product solution is reduced oxytocin [ 4‑Glu, 5‑Asp] impurity crude product solution is obtained after oxidation; the packing of high performance liquid phase chromatography is super water-resistant packing. The present invention uses the one-step method of on-line reverse-phase enrichment, reverse-phase salt conversion and reverse-phase purification to obtain a pure polypeptide product. The removal rate of impurities in the crude product solution is high, and the production process is optimized. The mobile phase in the salt stage is an aqueous solution, which is environmentally friendly and pollution-free, and the outflowing waste liquid can be directly treated for sewage and reused.

Description

technical field [0001] The invention relates to a method for refining oxytocin [4-Glu, 5-Asp] impurities. Background technique [0002] Oxytocin, also known as oxytocin, English name is Oxytocin, structural formula: [0003] [0004] The molecular formula is: C 43 h 66 N 12 o 12 S 2 , the molecular weight is 1007.2 [0005] Oxytocin is used to induce labor, oxytocin, postpartum and post-abortion uterine bleeding caused by uterine atony or poor abdominal contraction; understand placental reserve function (oxytocin provocation test); nasal drops can promote milk discharge. Oxytocin can indirectly stimulate the contraction of uterine smooth muscle, simulating the uterine contraction of normal childbirth, leading to cervical dilatation, and the uterine response to oxytocin gradually increases during pregnancy and reaches its peak at full term. Oxytocin can also stimulate the contraction of the smooth muscle of the mammary gland, which helps the milk to be discharged fr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/16C07K1/20
CPCC07K7/16
Inventor 江锡铭丁金国黄臻辉
Owner SPH NO 1 BIOCHEM & PHARMA CO LTD
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