Biological dehydrogenation method of androstenedione C1 and 2 loci
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A kind of androstenedione and biological technology, applied in the field of biological dehydrogenation of androstenedione C1, 2, can solve the problems of low conversion rate, complicated operation, high degradation rate, etc., and achieve fast conversion time, simple operation and high degradation low rate effect
Active Publication Date: 2020-01-07
HUNAN NORCHEM PHARMACEUTICAL CO LTD
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[0004]
In view of the above technical problems, the purpose of the present invention is to provide a fast, high-purity, simple Nocardia microbial conversion of 4-androstene-3,17-dione to prepare 1,4-androstene- The method of 3,17-dione can solve the problems of complex operation, low conversion rate and high degradation rate in the current dehydrogenation method of androstenedione
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[0031] Seed culture was carried out according to Example 1, the primary seed solution was inserted into the secondary seed medium for 10 hours, 2g4-AD was added to the cultured 100ml secondary seeds, and transformed for 72 hours, transformation conditions: 200rpm, 30±1°C. After the conversion, the sample was sent to the liquid phase, the normalized content of ADD was 82.19%, and the normalized content of 4-AD was 17.18%.
Embodiment 3
[0032] Embodiment 3 shake flask transformation
[0033] Carry out seed cultivation according to Example 1, insert the primary seed liquid into the secondary seed medium and cultivate for 16 hours, add 2 g of 4-AD crushed to 200 meshes into the cultivated 100 ml secondary seeds, transform for 72 hours, transformation conditions: 200 rpm, 30±1°C. After the conversion, the sample was sent to the liquid phase, the normalized content of ADD was 85.16%, and the normalized content of 4-AD was 14.49%.
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Abstract
The invention relates to production methods of steroid medicine intermediates, and specifically relates to a biological dehydrogenation method of androstenedione C1 and 2 loci. The biological dehydrogenation method of the androstenedione C1 and 2 loci comprises the following steps: taking 4-androstene-3,17-dione as a substrate and simple nocardia bacterium fluid as an enzyme source, adding soybeanoil at a concentration of 100-200 ml/L and tween-80 at a concentration of 0.1-2 g/L into a transformation system, and carrying out transformation reaction at 29-31 DEG C until completion of the reaction so as to obtain a reaction product; and then, separating and purifying the reaction product so as to obtain the 1,4-androdiene-3,17-dione. On the basis of adopting a common direct conversion method, the soybean oil and the tween-80 are added according to the biological dehydrogenation method of the androstenedione C1 and 2 loci, so that conversion rate can be greatly improved that the conversion rate can be up to 96% or above; and moreover, the product is low in degradation rate, high in reaction specificity, relatively fast in conversion time and high in product quality. In addition, sterile environment is not required that only routine outdoor operation is enough; and thus, the biological dehydrogenation method of the androstenedione C1 and 2 loci is easy to operate, and suitable forindustrial large-scale production.
Description
technical field [0001] The invention relates to a production method of a steroid drug intermediate, in particular to a method for biological dehydrogenation of the C1 and 2 positions of androstenedione. Background technique [0002] There are two kinds of dehydrogenation methods of C1 and 2 positions of steroidal compounds: chemical method and biological method. The traditional chemical method uses arsenic dioxide to dehydrogenate. The process is simple and the yield is considerable, but the product contains a certain amount of arsenic, which cannot meet the relevant standards. According to regulations, the biological method has the characteristics of strong specificity, rapid response and high yield, so the biological method has gradually replaced the chemical method in recent years. [0003] The reaction process of androstenedione C1,2 position dehydrogenation is as follows figure 1 shown. Androstenedione is dehydrogenated at the C1 and 2 positions. There are currently f...
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