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Method for purifying rapamycin from broth

A technology of rapamycin and fermentation liquid, which is applied in the field of biomedicine, can solve the problems of low yield of rapamycin, lengthy and complicated extraction and purification process, and low yield, achieve high practical value, and simplify purification steps , The effect of increasing the yield

Active Publication Date: 2012-05-09
LUNAN PHARMA GROUP CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the productive rate of utilizing this method to purify rapamycin is not high, in a specific example, it can be known that the final pure product of 11kg of rapamycin fermented liquid only has 6g, takes 3g pure product and further purifies and crystallizes and obtains 2.5 g
[0007] From the extraction and purification processes of the above several patent documents, we can know that the purity of rapamycin has increased to more than 99.8%, or the content of simple impurities and total impurities has also been reduced to below 0.15% and 1.2%, but its The extraction and purification process is tedious and complicated, and the yield is very low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] a. extract the mycelium obtained after filtering the fermented liquid (25L, titer is 1.1g / L) with 8 times the volume of acetone, extract 3 times to obtain the extract, and vacuum concentrate to obtain the concentrated extract;

[0055] b. Use 0.6 times the volume of ethyl acetate to extract the supernatant from the fermentation broth and filter it, control the temperature of the fermentation supernatant to 25°C and pH to 7.1, separate the ethyl acetate phase to obtain the extract, and concentrate in vacuo to obtain a concentrated extract ;

[0056] c. the concentrated extraction solution in step a and the concentrated extraction solution in step b are combined to obtain a concentrated solution, and added to a bed layer equipped with a macroporous resin DiaionHP20 for adsorption, wherein the volume ratio of DiaionHP20 to the concentrated solution is 1: 12. Wash with 2 times the volume of DiaionHP20 in an aqueous solution containing 45% acetone, control the temperature of...

Embodiment 2

[0063] a. Extract the mycelia obtained after filtering the fermented liquid (5,000 L, titer 0.99 g / L) with 10 times the volume of ethanol, extract 4 times to obtain the extract, and concentrate in vacuo to obtain the concentrated extract;

[0064] b. Use 1 volume of ethyl acetate to extract the supernatant obtained after the fermentation broth is filtered, control the temperature of the fermentation supernatant to 28°C and pH to 5.5, separate the ethyl acetate phase to obtain the extract, and concentrate in vacuo to obtain the concentrated extract liquid;

[0065] c. Combine the concentrated extraction solution in step a and the concentrated extraction solution in step b to obtain a concentrated solution, and add it to the bed layer equipped with macroporous resin X-5 for adsorption, wherein the volume of X-5 and the concentrated solution Ratio is 1: 15, washes with the aqueous solution containing 50% acetone of 4 times of X-5 volumes, controls the temperature of the aqueous s...

Embodiment 3

[0072] a. Extract the mycelia obtained after filtering the fermented liquid (30,000L, titer 0.91g / L) with 5 times the volume of acetone, extract 2 times to obtain the acetone extract, and vacuum concentrate to obtain the concentrated extract;

[0073] b. Use 0.2 times the volume of ethyl acetate to extract the supernatant obtained after the filtration of the fermentation broth, control the temperature of the fermentation supernatant to 22°C and pH to 6.5, separate the ethyl acetate phase to obtain the extract, and concentrate in vacuo to obtain a concentrated extract liquid;

[0074] c. the concentrated extraction solution in step a and the concentrated extraction solution in step b are combined to obtain concentrated solution, and join the bed layer that macroporous resin SP825 is housed and adsorb, wherein the volume ratio of SP825 and concentrated solution is 1: 8. Wash with 1 times the volume of SP825 in an aqueous solution containing 40% acetone, control the temperature o...

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Abstract

The invention belongs to the biomedicine technical field, concretely relates to a method for purifying rapamycin from broth. In order to raise the purity of rapamycin, reduce the impurity content in rapamycin and increase the yield of rapamycin, the invention uses mycelium extraction, broth extraction, macroporous resin adsorption, silicagel column chromatography, cooling crystallization and recrystallization to extract and purify the rapamycin pure product from the broth, the purity is greater than 99.3%, the single impurity is less than 0.10% and the yield is increased to 53-60%. According to the invention, the rapamycin purity is raised, the impurity content is reduced, the yield is enhanced simultaneously, the operation is convenient, the technology is simple, and the method for purifying rapamycin from broth has high utility value in large industrial production.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for purifying rapamycin from fermentation broth. Background technique [0002] Rapamycin (RPM, RAPA), also known as Sirolimus, is a triene macrolide compound composed of 31-membered rings, containing special α, β-diketopipexamine Molecularly masked hemicarboxyketones with the formula C 51 h 79 o 13 , the molecular weight is 991KD. It was isolated from Strepomyces hygroscopicus in 1975 by the Ayerst Research Institute in Canada. [0003] Vezinia found that it has anti-fungal effect in 1975, and later Tanaka and others found that it is a promising immunosuppressant and has anti-tumor effect. Studies by Tanaka et al. have shown that rapamycin has a strong anti-graft rejection reaction, can synergize with cyclosporine to prolong the survival time of grafts, reverse ongoing transplant rejection, and prevent and treat experimental autohemolytic anemia rol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D498/18
Inventor 赵志全赵丽丽
Owner LUNAN PHARMA GROUP CORPORATION
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