Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Large-scale separation and purification method of teriparatide

A technology for separation and purification of teriparatide, which is applied in the field of large-scale separation and purification of teriparatide by reversed-phase high performance liquid chromatography, can solve problems such as unfavorable industrial production, unstable yield, peptide degradation, etc. The effect of large-scale industrial production, high production yield and high product purity

Inactive Publication Date: 2016-11-30
ZHEJIANG PEPTITES BIOTECH CO LTD
View PDF5 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the product is dissolved in 60% phosphoric acid aqueous solution in the salting-out step, it is easy to cause peptide degradation, resulting in unstable yield
[0007] Patent CN102731643A discloses a purification method of teriparatide, using water as solvent, then using 0.2% TFA / acetonitrile as mobile phase to use C18 column to purify, and then using 0.2% acetic acid / acetonitrile C18 column to transfer salt, although it can get Higher purity teriparatide acetate product, but the total yield is only about 20%, which is not conducive to industrial production
[0008] In order to solve the problem of low yield in the prior art, improve the purification yield of teriparatide acetate, and reduce the production cost, further research on the purification method is required

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] 1. Sample treatment: Dissolve each gram of crude teriparatide in 50ml of acetonitrile aqueous solution with a volume ratio: acetonitrile: water = 30:70, sonicate, and after the sample is completely dissolved, filter it with a filter membrane with a pore size of 0.45 μm and collect the filtrate stand-by.

[0032] 2. Purification: Purification conditions: Chromatographic column: DAC-CXTH200 dynamic axial pressurized column with octadecylsilane bonded silica gel as the stationary phase, column diameter and packing length: 20*25cm. Mobile phase A: 0.2% ammonium acetate aqueous solution by mass percentage, adjust the pH value to 5.0 with acetic acid; phase B: acetonitrile. The flow rate is 700~1000ml / min. The detection wavelength is 220nm. Gradient: B%: 20~70% (60min). The injection volume is 60g.

[0033] Purification process: Equilibrate the chromatographic column with mobile phase A and load the sample, with a sample volume of 3L sample solution. The linear gradient ...

Embodiment 2

[0036] 1. Sample treatment: Dissolve each gram of crude teriparatide in 50ml of acetonitrile aqueous solution with a volume ratio: acetonitrile: water = 30:70, sonicate, and after the sample is completely dissolved, filter it with a filter membrane with a pore size of 0.45 μm and collect the filtrate stand-by.

[0037] 2. Purification: Purification conditions: Chromatographic column: DAC-CXTH200 dynamic axial pressurized column with octadecylsilane bonded silica gel as the stationary phase, column diameter and packing length: 20*25cm. Mobile phase A: 0.4% ammonium acetate aqueous solution by mass percentage, adjust the pH value to 5.5 with acetic acid; phase B: acetonitrile. The flow rate is 700~1000ml / min. The detection wavelength is 220nm. Gradient: B%: 25~75% (60min). The injection volume is 80g.

[0038] Purification process: equilibrate the chromatographic column with mobile phase A and then load the sample, with a sample volume of 4L sample solution. The linear grad...

Embodiment 3

[0041] 1. Sample treatment: Dissolve each gram of crude teriparatide in 50ml of acetonitrile aqueous solution with a volume ratio: acetonitrile: water = 30:70, sonicate, and after the sample is completely dissolved, filter it with a filter membrane with a pore size of 0.45 μm and collect the filtrate stand-by.

[0042] 2. Purification: Purification conditions: Chromatographic column: DAC-CXTH200 dynamic axial pressurized column with octadecylsilane bonded silica gel as the stationary phase, column diameter and packing length: 20*25cm. Mobile phase A: 0.6% ammonium acetate aqueous solution by mass percentage, adjust the pH value to 6.0 with acetic acid; phase B: acetonitrile. The flow rate is 700~1000ml / min. The detection wavelength is 220nm. Gradient: B%: 30~80% (60min). The injection volume is 100g.

[0043] Purification process: equilibrate the chromatographic column with mobile phase A and load the sample, with a sample volume of 5L sample solution. After linear gradie...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a large-scale separation and purification method of teriparatide by reversed high performance liquid chromatography. The method comprises the following steps: dissolving a teriparatide crude product in 30% acetonitrile aqueous solution, carrying out ultrasonic treatment, filtering with a filter membrane, and fetching a filtrate for later use; separating and purifying the filtered crude product by a high performance liquid chromatography instrument; letting a concentrated solution pass through a reversed-phase column for conversion to acetate; carrying out vacuum decompression rotary-evaporation concentration on a product with 99% purity after conversion to acetate, freeze-drying to obtain a powdery product, and detecting qualified so as to obtain the final product. The method is convenient to operate and excessive generating equipment is not required. Product purity is high and yield is high. Product purity reaches 99% and above, purification recovery rate reaches 60% and above. The technology can meet requirements of large-scale industrial production.

Description

technical field [0001] The invention relates to a method for large-scale separation and purification of teriparatide, in particular to a method for large-scale separation and purification of teriparatide by reversed-phase high-performance liquid chromatography. Background technique [0002] Teriparatide is a synthetic 34-peptide, which is a 1-34 amino acid fragment of human parathyroid hormone PTH. The amino acid sequence is as follows: [0003] H-Ser1-Val2-Ser3-Glu4-Ile5-Gln6-Leu7-Met8-His9-Asn10-Leu11-Gly12-Lys13-His14-Leu15-Asn16-Ser17-Met18-Glu19-Arg20-Val21-Glu22-Trp23-Leu24- Arg25-Lys26-Lys27-Leu28-Gln29-Asp30-Val31-His32-Asn33-Phe34-OH [0004] This fragment is the biologically active N-terminal region of endogenous parathyroid hormone PTH containing 84 amino acids. The immunological and biological characteristics of this product are exactly the same as those of endogenous parathyroid hormone PTH and bovine parathyroid hormone PTH (bPTH). [0005] Teriparatide is t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/635C07K1/20C07K1/18C07K1/16
CPCC07K14/635
Inventor 刘志国李雪豪纪东亮秦德志
Owner ZHEJIANG PEPTITES BIOTECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com