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Method for removing impurities in pentostatin fermentation liquid

A pentostatin and fermentation broth technology, applied in chemical instruments and methods, preparation of sugar derivatives, sugar derivatives, etc., can solve the problems of complicated operation steps, large usage of organic solvents, serious environmental pollution, etc. Simple steps, good separation effect and convenient operation

Inactive Publication Date: 2012-05-16
SHANGHAI INST OF PHARMA IND
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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 overcome the defects of cumbersome operation steps, large amount of organic solvent used and serious environmental pollution in the existing pretreatment method for pretreatment of pentostatin fermented liquid, and to provide a method with convenient operation and high process steps. Simple, no special addition of organic solvents, a method that can effectively remove impurities in pentostatin fermentation broth

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] After high-speed centrifugation of the pentostatin fermentation broth, take the supernatant, adjust the pH to 3.0 with 1mol / L hydrochloric acid, and filter the supernatant with a hollow fiber ultrafiltration membrane with a molecular weight cut-off of 20,000 at 30°C and 0.3Mpa. The volume is 49.5L / h, the yield of the ultrafiltration membrane is 98.3%, and the concentration factor is about 1.5 times. The soluble protein, high polymer, medium, mycelium and other impurities in the supernatant are removed, and the filtrate is clarified good degree;

[0034] Then use nanofiltration membrane to further purify the ultrafiltrate, first adjust the pH of the filtrate to 5.0 with 1mol / L sodium hydroxide, then use the nanofiltration membrane with a cut-off molecular weight of 500 to filter at 40°C and 0.3Mpa, the average membrane pass The volume is 30.7L / h, the concentration factor is about 2 times, the filtrate is collected, the yield is about 82.1%, and some impurities such as po...

Embodiment 2

[0037] After high-speed centrifugation of the pentostatin fermentation broth, take the supernatant, adjust the pH to 4.0 with 1mol / L hydrochloric acid, and then filter the supernatant with a hollow fiber ultrafiltration membrane with a molecular weight cut-off of 5000 at 35°C and 0.2Mpa. The membrane flux is 45.8L / h, the yield of the ultrafiltration membrane is 94.8%, and the concentration factor is about 1.5 times. The soluble protein, high polymer, culture medium, mycelium and other impurities in the supernatant are removed, and the filtered out Liquid clarity is better;

[0038] Then use the nanofiltration membrane to further purify the ultrafiltrate, first adjust the pH of the filtrate to 6.0 with 1mol / L sodium hydroxide, and then filter it with a nanofiltration membrane with a cut-off molecular weight of 500 at 45°C and 0.25Mpa. The volume is 30.5L / h, the concentration factor is about 2 times, and the filtrate is collected with a yield of about 82.8%, and some impurities ...

Embodiment 3

[0041] After high-speed centrifugation of the pentostatin fermentation broth, take the supernatant, first adjust the pH to 5.0 with 1mol / L hydrochloric acid, and then filter the supernatant at 30°C and 0.3Mpa with a hollow fiber ultrafiltration membrane with a molecular weight cut-off of 8000. The average membrane flux is 47.6L / h, the yield of the ultrafiltration membrane is 96.4%, and the concentration factor is about 1.5 times. The soluble protein, high polymer, medium, mycelium and other impurities in the supernatant are removed, and the filtration The clarity of the liquid is better;

[0042] Then use the nanofiltration membrane to further purify the ultrafiltrate, first adjust the pH of the filtrate to 4.5 with 1mol / L sodium hydroxide, and then filter it with a nanofiltration membrane with a cut-off molecular weight of 500 at 60°C and 0.35Mpa. The volume is 26.4L / h, the concentration factor is about 2 times, the filtrate is collected, the yield is about 79.5%, and some im...

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Abstract

The invention discloses a method for removing impurities in a pentostatin fermentation liquid, which comprises the following steps: passing a centrifugalized pentostatin fermentation liquid supernatant through an ultrafiltration membrane of which the Molecular Weight Cut Off is 5000-20000, passing the ultrafiltrate through a nanofiltration membrane of which the Molecular Weight Cut Off is 500-2000, and passing the nanofiltrate through a nanofiltration membrane of which the Molecular Weight Cut Off is 100-200, thereby obtaining a filtrate. The method is convenient to operate, has the advantages of simple technical steps and favorable separating effect, does not need to especially add any organic solvent, and is suitable for industrial production.

Description

technical field [0001] The invention relates to a method for removing impurities in pentostatin fermentation liquid. Background technique [0002] Pentostatin was isolated from the culture medium of Streptomyces antibioticus by Woo et al. in 1974. It is a strong adenosine deaminase (ADA) inhibitor. Pentostatin has a high affinity with ADA, can bind tightly to ADA, inhibit the activity of ADA, and increase the level of deoxyadenosine triphosphate (dATP) in cells. dATP blocks DNA synthesis by inhibiting ribonucleotide reductase. In addition, pentostatin inhibits RNA synthesis and enhances DNA damage. In February 1998, the US FDA officially approved the marketing of pentostatin for injection (trade name Nipent), which is mainly used for stem cell leukemia (Hairy cell leukemia, HCL), chronic lymphocytic leukemia (Chronic lymphocytic leukemia, CLL) and mycosis fungoides (Mycosis fungoides) treatment. [0003] However, there are not many domestic and foreign studies on the sepa...

Claims

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Application Information

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IPC IPC(8): C07H19/23C07H1/08
Inventor 李继安刘文静卢亮
Owner SHANGHAI INST OF PHARMA IND
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