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Method for refining bleocin

A refining method and bleomycin technology, applied in the refining field of bleomycin, can solve the problems of low degree of automation and low yield, and achieve the effects of improving the degree of automation and reducing costs

Inactive Publication Date: 2009-06-24
SHANGHAI INST OF PHARMA IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing extraction method is to obtain the crude bleomycin through crude extraction, and then refine it by gel chromatography. The yield of this refining method is low (generally around 70%), and the degree of automation is not high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] use C 18 Silica gel column (Hypersil filler, 20mm x 250mm, 10um), detection wavelength 254nm. The crude bleomycin was dissolved in water (200mg dissolved in 10ml of water, 1ml for each injection). Mobile phase ratio: Dissolve 1.0 g of ammonium sulfate in 1000 ml of water, adjust the pH to 3.5 with acetic acid, and use methanol as the organic phase. The flow rate was 15ml / min, and the proportion of the organic phase increased from 15% to 30% within 30 minutes, and maintained for 10 minutes. The components of bleomycin could be separated well, and the yield was 87.2%.

Embodiment 2

[0023] use C 8 Silica gel column (Hypersil filler, 20mm x 250mm, 10um), detection wavelength 254nm. The crude bleomycin was dissolved in water (100mg dissolved in 10ml water, 2ml for each injection). Mobile phase ratio: Dissolve 5.0 g of potassium sulfate in 1000 ml of water, adjust the pH to 4.5 with acetic acid, and use acetonitrile as the organic phase. The flow rate is 16ml / min, the proportion of the organic phase rises from 12% to 28% within 30min, and is maintained for 10min. The components of bleomycin can be separated well, and the yield is 88.7%.

Embodiment 3

[0025] use C 18 Silica gel column (Hypersil filler, 20mm x 250mm, 10um), detection wavelength 280nm. The crude bleomycin was dissolved in water (150mg dissolved in 10ml of water, 1ml for each injection). Mobile phase ratio: Dissolve 0.5 g of sodium sulfate in 1000 ml of water, adjust the pH to 5 with acetic acid, and use methanol as the organic phase. The flow rate was 13ml / min, the proportion of the organic phase rose from 18% to 35% within 35min, and was maintained for 10min. The components of bleomycin could be separated well, and the yield was 85.3%.

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PUM

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Abstract

The invention discloses a refining method for bleocin, comprising purifying the crude product of bleocin by preparative HPLC technology; wherein, the immobile phase of a preparation column is silica gel column of C8 or C18, the mobile phase is mixed liquid of sulphate aqueous solution with pH value between 3.5 and 5 and polar organic solvent. In the method, the preparative HPLC technology is adopted to increase the yield (85% generally) and the degree of automation of A2 and A3 which are the active ingredients of bleocin and greatly lower cost, thus can be applied to industrial production.

Description

technical field [0001] The invention relates to a method for refining bleomycin. Background technique [0002] Due to the deterioration of the environment and unhealthy living habits, the incidence of tumors continues to increase worldwide. Since 1996, more than 10.3 million cancer patients have been newly diagnosed every year in the world. By the end of 1999, the total number of cancer patients in the world has exceeded 40 million. The World Health Organization reported in 2001 that the incidence and mortality of cancer in the world increased by 22% compared with 1990, and will increase by about 50% in the next 20 years. [0003] Bleomycin (Bleomycin) is a broad-spectrum antibacterial and antitumor antibiotic with a unique structure and action. It is effective for a variety of tumor diseases, and does not cause leukopenia, does not inhibit the immune function of the body, and does not damage the hematopoietic system. The mechanism by which bleomycin kills tumor cells is t...

Claims

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Application Information

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IPC IPC(8): C07K9/00C07K1/22
Inventor 刘垚李继安陈代杰
Owner SHANGHAI INST OF PHARMA IND
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