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Method for preparing acarbose

A kind of technology of acarbose and seeds, applied in the field of preparation of α-glucosidase inhibitor active ingredients

Inactive Publication Date: 2009-12-16
SHANGHAI INST OF PHARMA IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, high concentration of glucose will have a feedback inhibition effect on the secondary metabolism of motile actinomycetes

Method used

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  • Method for preparing acarbose
  • Method for preparing acarbose
  • Method for preparing acarbose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Inoculate the slant of Actinomyces mobilis on the slant medium, and cultivate it at 28°C for 6 days; slant medium: starch hydrolyzate 30mg / ml; peptone 5mg / ml; dipotassium hydrogen phosphate 0.6mg / ml; magnesium sulfate 0.6 mg / ml; agar 20mg / ml; pH before digestion = 7.0; medium loading 20%;

[0031] (2) Select the full-grown slant and inoculate it into the seed medium, and cultivate it for 44 hours at 28° C. and 200 rpm; seed medium: starch 10 mg / ml; glycerin 20 mg / ml; soybean cake powder (hot fried) 20 mg / ml; calcium carbonate 2mg / ml; pH before digestion = 6.8; 750ml shake flask seed medium capacity is 90ml;

[0032] (3) Fermenter culture, after 40 hours, take samples every 6 hours to detect the content of glucose, maltose and amino nitrogen. ml and amino nitrogen 0.13mg / ml; at the same time, the unit of acarbose was detected by high-pressure liquid phase, and the unit was 2840 μg / ml after 8 days, and the content of component C was 11.6 μg / ml (accounting for 0.41wt ...

Embodiment 2

[0034] (1) Inoculate the slant of Actinomyces mobilis on the slant medium, and cultivate it at 25°C for 7 days; slant medium: starch hydrolyzate 50mg / ml; peptone 4mg / ml; dipotassium hydrogen phosphate 0.4mg / ml; magnesium sulfate 0.4 mg / ml; agar 15mg / ml; pH before digestion = 7.0; slant medium load 12%;

[0035] (2) Select the full-grown slant and inoculate it into the seed medium, and cultivate it for 48 hours at 25°C and 240rpm; seed medium: starch 8mg / ml; glycerol 15mg / ml; soybean cake powder (hot fried) 20mg / ml; calcium carbonate 2mg / ml; pH before digestion = 6.8; 750ml shake flask seed medium capacity is 150ml;

[0036](3) Fermentation tank culture, after 70 hours, take samples and detect glucose, maltose and ammonium chloride every 12 hours, add glucose, maltose and ammonium chloride according to the measurement results, so that the content is controlled at 10 mg / ml of glucose and 25 mg / ml of maltose ml and amino nitrogen 0.05mg / ml; at the same time, the unit of acarbose...

Embodiment 3

[0038] (1) Inoculate the slant of Actinomycetes mobilis on the slant medium, and cultivate it at 25°C for 6 days; slant medium: starch hydrolyzate 30mg / ml; peptone 6mg / ml; dipotassium hydrogen phosphate 0.5mg / ml; magnesium sulfate 0.5 mg / ml; agar 16mg / ml; pH before digestion = 7.0; slant medium load 15%;

[0039] (2) Select the full-grown slant and inoculate it into the seed medium, and cultivate it for 46 hours at 25°C and 220rpm; seed medium: starch 12mg / ml; glycerin 25mg / ml; soybean cake powder (hot fried) 15mg / ml; calcium carbonate 2mg / ml; pH before digestion = 6.8; 750ml shake flask seed medium capacity is 100ml;

[0040] (3) Fermentation tank culture, after 48 hours, take samples every 6 hours to detect the content of glucose, maltose and amino nitrogen, and add glucose, maltose and aspartic acid according to the measurement results, so that the content is controlled at 8 mg / ml of glucose and 23 mg of maltose / ml and amino nitrogen 0.15mg / ml; at the same time, the unit ...

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Abstract

The invention discloses a method for preparing acarbose, which comprises the following steps: (1) actinoplane slants are inoculated in a slant culture medium for slant culture; (2) slants with full growth are selected and inoculated in a seed culture medium for seed culture; (3) fermentation cylinder culture is carried out, and glucose and maltose are added into a fermentation culture medium in afluid mode in a culture process. The method is characterized in that a nitrogen source is also added in the fluid mode in the step (3); and the content of amino nitrogen in the culture medium after adding the nitrogen source in the fluid mode is controlled to be smaller than or equal to 0.15 mg / ml except 0 mg / ml. The method overcomes the defect of difficult separation of components C which are impurities in the prior art, replenishes the nitrogen source in the process of fermentation, controls the content of the nitrogen source and can reduce the formation of the impurities C while greatly improving the yield of the acarbose.

Description

technical field [0001] The invention relates to a preparation method of an alpha-glucosidase inhibitor active ingredient, in particular to a preparation method of acarbose. Background technique [0002] Acarbose (acarbose) is an effective α-glucosidase inhibitor, its molecular structure is shown in the following formula 1, and it is widely used clinically to treat non-insulin-dependent type II diabetes. The fermentation production of acarbose must optimize the fermentation conditions to increase the fermentation unit and reduce the cost. However, during the fermentation and production of actinomycetes (Actinoplanessp.), a series of isomers are generated: components A, B, C, D, E, F, G and H (European Pharmacopoeia 5.1P2873-2874) . European Pharmacopoeia clearly stipulates the limits of impurities: component A<0.6%, B<0.5%, C<1.5%, D<1.0%, E<0.2%, F<0.3%, G<0.3%, H<0.2%, Other impurities are less than 0.2%. Wherein impurity component A and componen...

Claims

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Application Information

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IPC IPC(8): C12P19/26C12N1/20C12R1/045
Inventor 胡海峰张琴
Owner SHANGHAI INST OF PHARMA IND
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