Unlock instant, AI-driven research and patent intelligence for your innovation.

A chromatographic method for effectively improving the purification yield of synthetic peptides

A purification method and peptide synthesis technology, which is applied in the field of preparation and purification of liraglutide, can solve the problems of small sample size, low purity, and large single impurities of peptide products, and achieve the goal of improving the total yield of purification and reducing purification steps Effect

Active Publication Date: 2021-03-02
SINOPEP ALLSINO BIOPHARMACEUTICAL CO LTD +1
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The specific implementation principle is to increase the packing height of a single chromatographic column, improve the separation of impurities in the synthetic peptide by a single chromatographic column, and improve the sample purification yield. Technical problems such as low purity and low efficiency; the above strategies are suitable for dynamic axial compression columns

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

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A chromatographic method for effectively improving the purification yield of synthetic peptides
  • A chromatographic method for effectively improving the purification yield of synthetic peptides
  • A chromatographic method for effectively improving the purification yield of synthetic peptides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1 Purification of liraglutide

[0071] 1.1 Column bed height: 300mm

[0072] Take the crude product of target liraglutide

[0073] Sample processing: 150.0 g of liraglutide sample was dissolved in an aqueous acetonitrile solution, and filtered through a 0.22 μm filter after complete dissolution. The filtered aqueous solution of crude liraglutide peptide was collected for use.

[0074] 1st step HPLC purification

[0075] Chromatographic conditions: use octadecyl or octadecyl bonded silica gel filler stationary phase (100mm×250mm, 8-20μm) as the chromatographic column; use 0.1% ammonium sulfate (take 1000ml of water, add 1ml of ammonium sulfate, mix well, use Ammonia water to adjust pH to 3.3) as mobile phase A; acetonitrile as mobile phase B; flow rate of 20 mL per minute, multiple gradient elution, detection wavelength of 230 nm; single-needle loading of 15.0 g.

[0076] The following table elution gradients were used for elution.

[0077]

[0078] Fract...

Embodiment 2

[0110] Example 2 Purification of semaglutide

[0111] 2.1 Column bed height: 300mm

[0112] Take crude somaglutide

[0113] Sample treatment: Dissolve the sample containing 12.0 g of semaglutide in an aqueous acetonitrile solution, and filter it with a 0.22 μm filter after the complete dissolution. The filtered crude semaglutide aqueous solution was collected for use.

[0114] 1st step HPLC purification

[0115] Chromatographic conditions: use octadecyl or octadecyl bonded silica gel filler stationary phase (50mm×300mm, 10μm) as the chromatographic column; use 0.1% TFA (take 1000ml of water, adjust the pH value to 2.3 with ammonia water) as the mobile phase A ; Take acetonitrile as mobile phase B; flow rate is 20mL per minute, multiple gradient elution, detection wavelength is 230nm; single needle loading is 3.0g.

[0116] The following table elution gradients were used for elution.

[0117]

[0118] Fractions of semaglutide samples with a purity greater than 90.0% were ...

Embodiment 3

[0152] Take crude liraglutide

[0153] Sample processing: 150.0 g of liraglutide sample was dissolved in an aqueous acetonitrile solution, and filtered through a 0.22 μm filter after complete dissolution. The filtered aqueous solution of crude liraglutide peptide was collected for use.

[0154] 1st step HPLC purification

[0155] Chromatographic conditions: use octadecyl or octadecyl bonded silica gel filler stationary phase (100mm×300mm, 8-20μm) as the chromatographic column; use 0.1% ammonium sulfate (take 1000ml of water, add 1ml of ammonium sulfate, mix well, use Ammonia water to adjust pH to 3.3) as mobile phase A; acetonitrile as mobile phase B; flow rate of 20 mL per minute, multiple gradient elution, detection wavelength of 230 nm; single-needle loading of 15.0 g.

[0156] The following table elution gradients were used for elution.

[0157]

[0158] Fractions of liraglutide samples with a purity greater than 95.0% were collected. Part of the acetonitrile was re...

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

PropertyMeasurementUnit
heightaaaaaaaaaa
heightaaaaaaaaaa
heightaaaaaaaaaa
Login to View More

Abstract

The invention belongs to the field of medicine preparation, and relates to a chromatographic method for improving the purification yield of synthetic polypeptides, in particular to a method for improving the purification yield of liraglutide, and the method can effectively remove substances whose physical and chemical characteristics are less different from those of the target Impurities, such as achiral enantiomer impurities, are especially suitable for the purification of samples with complex impurity profiles; at the same time, they can also solve technical problems such as small sample loading, large single impurities, low purity, and low efficiency of peptide products.

Description

technical field [0001] The invention belongs to the field of pharmaceutical preparation, in particular to a preparation and purification method of liraglutide. Background technique [0002] Liraglutide is the first long-acting human glucagon-like peptide-1 (GLP-1) analog developed by Novo Nordisk, which shares 97 homology with GLP-1. %, its peptide sequence is: H-His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-Lys(γ -Glu-Palmitoyl)-Glu-Phe-Ile-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OH; liraglutide can lower blood sugar, promote islet cell regeneration, slightly prolong gastric emptying, etc. It has a wide range of application prospects. As a new generation of incretin-based hypoglycemic drugs, liraglutide not only has a long duration of action, but also fully retains many physiological activities of natural GLP-1, which can safely and effectively lower blood sugar and may have a variety of cardiovascular hazards. factors play a protective role. Lirag...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/605C07K1/36C07K1/34C07K1/20B01D15/18B01D15/32B01D15/42
CPCB01D15/1871B01D15/325B01D15/426C07K14/605
Inventor 赵呈青谷海涛肖英
Owner SINOPEP ALLSINO BIOPHARMACEUTICAL CO LTD