Cetrorelix purification and separation method

A technology of purification and separation and cetrorelix, which is applied in the field of polypeptide drug preparation, can solve problems such as affecting product yield and difficult separation, and achieve the effect of reducing production cost, good yield and high purity

Inactive Publication Date: 2017-11-03
ZHEJIANG PEPTITES BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the process of producing cetrorelix, we also found that deamidated cetrorelix impurities and acetylated cetrorelix impurities are difficult to separate, which seriously affects the product yield

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Take 50 g of the crude product of Cetrorelix, dissolve each gram of the crude product of Cetrorelix in 100 ml of 35v / v% acetonitrile aqueous solution, and perform ultrasonic treatment. After the crude product is completely dissolved, filter it with a filter membrane with a pore size of 0.45 μm, and collect the filtrate for later use .

[0036] 2. Purification: Reverse-phase chromatographic column: C18 is used as the stationary phase, and the diameter and packing length of the column are: 20*25cm. Mobile phase A: an aqueous solution of sodium dihydrogen phosphate with a mass concentration of 0.4 wt%, and adjust the pH value to 3.3 with phosphoric acid; phase B: acetonitrile. The flow rate is 800ml / min. The detection wavelength is 220nm. After equilibrating the reversed-phase column with mobile phase A, load the filtrate from step 1 onto the reversed-phase column. A gradient: B%: 22-62% (60 min) was used for elution, and the elution peaks were collected.

[0037] 3...

Embodiment 2

[0041] 1. Take 65g of the crude product of cetrorelix, dissolve each gram of the crude product of cetrorelix in 100ml of 35v / v% acetonitrile aqueous solution, and process it with ultrasonic waves. .

[0042] 2. Purification: Reverse-phase chromatographic column: C18 is used as the stationary phase, and the diameter and packing length of the column are: 20*25cm. Mobile phase A: an aqueous solution of sodium dihydrogen phosphate with a mass concentration of 0.8 wt%, and adjust the pH value to 3.8 with phosphoric acid; phase B: acetonitrile. Flow rate 1000ml / min. The detection wavelength is 220nm. After equilibrating the reversed-phase column with mobile phase A, load the filtrate from step 1 onto the reversed-phase column. Gradient: B%: 20-60% (60min) was used for elution, and the sample solution was collected.

[0043] 3. Concentration: Concentrate the collected target peptide solution with a purity of more than 99.5% by rotary evaporation under reduced pressure at no more th...

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Abstract

The invention discloses a cetrorelix purification and separation method. The method includes steps: dissolving a crude product of cetrorelix in acetonitrile water solution, and filtering through a filter membrane to obtain crude solution for standby application; adopting a mobile phase A for balancing a reversed phase column, loading the crude solution into the reversed phase column, performing gradient eluting for separation and purification, wherein the mobile phase A refers to sodium dihydrogen phosphate aqueous solution, and a mobile phase B refers to acetonitrile; subjecting target peptide solution with purity higher than 99.5% to vacuum rotary evaporation and concentration at a water temperature not higher than 38 DEG C; adopting acetic acid aqueous solution for balancing the reversed phase column, loading a sample of concentrated liquid into the reversed phase column, and adopting an acetic acid aqueous solution/acetonitrile system for salt conversion; subjecting the converted acetate and the target peptide solution with purity higher than 99.5% to vacuum rotary evaporation and concentration at a water temperature not higher than 38 DEG C, and performing freeze drying to obtain powdery cetrorelix. The obtained cetrorelix is high in purity and yield, meets industrial production requirements and has a high economic value and a promising application prospect.

Description

technical field [0001] The invention relates to the technical field of polypeptide medicine preparation, in particular to a method for purifying and separating cetrorelix. Background technique [0002] Cetrorelix, a synthetic 10-peptide, is a potent progesterone-releasing hormone-releasing hormone (LHRH) receptor antagonist that controls ovarian stimulation, prevents premature expulsion of immature follicles, and aids conception. And can be used for the treatment of breast cancer and gynecological cancer, endometriosis, precocious puberty, uterine fibroids, ovarian androgen excess and premenstrual syndrome. Due to its own physical properties, cetrorelix is ​​easily decomposed and destroyed by enzymes in the gastrointestinal tract, and cannot be taken orally. Therefore, it is often used as a cetrorelix preparation for injection, absorbed through subcutaneous or intramuscular injection, and rapidly exerts its effect. [0003] The sequence of cetrorelix is ​​as follows: [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/23C07K1/20
CPCC07K7/23
Inventor 刘志国陈晓航李雪豪
Owner ZHEJIANG PEPTITES BIOTECH CO LTD
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