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Method for preparing vasopressin tannate

A technology of vasopressin and tannic acid, applied in the direction of oxytocin/vasopressin, chemical instruments and methods, specific peptides, etc., can solve the problems of complex preparation process and low yield, and achieve the effect of optimizing the production process

Inactive Publication Date: 2015-06-03
QINGDAO KANGYUAN PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a preparation method of vasopressin tannin in order to overcome the defects of complex preparation process and low yield of vasopressin tannin in the prior art

Method used

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  • Method for preparing vasopressin tannate
  • Method for preparing vasopressin tannate
  • Method for preparing vasopressin tannate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 (HPLC method to detect the purity of crude vasopressin raw material and purified intermediate solution)

[0032] Instrument: Waters2695 / 2489 high performance liquid chromatography

[0033] Separation column: Agilent XDB-C184.6×250mm, 5μm

[0034] Mobile phase: A is 0.1% TFA aqueous solution in volume percentage, B is 0.1% TFA-50% acetonitrile aqueous solution in volume percentage

[0035] Flow rate is 1.0mL / min, detection wavelength is 214nm, room temperature detection,

[0036] The elution gradient is shown in Table 1 below, and the percentages are volume percentages.

[0037] Table 1 Mobile phase elution ladder

[0038] Elution step Elution time eluent 1 0~5min 95%A+5%B 2 5~25min 95%A+5%B→50%A+50%B 3 25~30min 100％B 4 30~35min 95%A+5%B

Embodiment 2

[0039] Embodiment 2 (75mm inner diameter L&L4003 preparative column packing)

[0040] Using Load&Lock dynamic axial compression and static locking technology, the filler is styrene-divinylbenzene copolymer (reversed phase packing Agilent PLRP-S), the pore size is 10nm, the particle size is 10μm, and the column packing density is 0.33g / mL. The bed pressure is 650psi, Varian chromatographic packing system is used, 370g dry powder filler, 2L methanol is stirred and homogenized, poured into a 75mm inner diameter L&L4003 preparative column, the compression ratio is 3:1, the carrier gas is N2, and the carrier gas pressure is adjusted so that the oil pressure gauge pressure 2000psi, dynamic axial compression to a column bed height of 26cm, used as a preparative column for reverse-phase purification and reverse-phase desalting schemes.

Embodiment 3

[0041] Embodiment 3 (reverse phase purification of vasopressin crude product raw material)

[0042] Instrument: Varian SD-1 high pressure liquid phase preparation system

[0043] Chromatographic column: self-packed preparative column Load&Lock400375×260mm in Example 2, PLRP-S10μm10nm

[0044] Mobile phase: A is 0.1% trifluoroacetic acid aqueous solution by volume, B is 0.1% trifluoroacetic acid acetonitrile solution by volume, and the crude vasopressin is dissolved in 5% acetonitrile aqueous solution as solvent to prepare a 10g / L solution , after filtration and clarification, the sample liquid is mobile phase C, and its HPLC purity measured by the method of Example 1 is 88.13%, and the retention time is 14.40min.

[0045] The reverse-phase purification conditions of this example are as follows: flow rate 200mL / min, detection at 220nm, purification and elution gradient as shown in the table below, and the main peak of the target with a retention time of 48-54min is collected a...

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Abstract

The invention discloses a method for preparing vasopressin tannate. The preparation method of the vasopressin comprises the following step: sequentially carrying out reverse-phase purification and reverse-phase desalination on a vasopressin crude product solution by a high performance liquid chromatography, so as to obtain the vasopressin tannate, wherein packing of the high performance liquid chromatography, is a polystyrene-divinylbenzene (PS-DVB) copolymer. By combination of reverse-phase purification and reverse-phase desalination, the latest application of the polymer packing polystyrene-divinylbenzene is designed; and the vasopressin tannate can be prepared on a large scale.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals. More specifically, the present invention relates to a method for the preparation of vasopressin tannate. Background technique [0002] Vasopressin tannin is a synthetic polypeptide composed of nine amino acid residues, and its chemical structure is L-Cys-L-Tyr-L-Phe-L-Gln-L-Asn-L-Cys-L-Pro- L-Arg-L-Gly(1→6)-disulfide bond tannate, the theoretical molecular weight of vasopressin is 1084.24. Vasopressin tannin is an antidiuretic hormone drug, which can promote the reabsorption of water in the distal renal tubule and collecting duct and has an antidiuretic effect. Its preparation is clinically used for the treatment of diabetes insipidus. [0003] At present, most of the common purification methods for peptide drugs on the market use preparative high-performance liquid chromatography, which is the most effective method for obtaining high-purity peptide target molecules. Generally, the purificati...

Claims

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Application Information

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IPC IPC(8): C07K7/16C07K1/20
Inventor 刘冠男李静洁孙延年
Owner QINGDAO KANGYUAN PHARMA
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