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Preparation method for creatine phosphate sodium

A technology of sodium creatine phosphate and creatine, which is applied in the chemical field, can solve the problems of reducing drug safety, increasing the difficulty of purification, and low use efficiency, and achieves the effects of high synthesis efficiency, improved stability, and good safety

Active Publication Date: 2015-12-09
XINAN PHARMA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the disadvantages of the free enzyme synthesis method are: the enzyme can only react once, and the use efficiency is low; after the enzyme reaction is completed, it remains in the reaction solution, which not only increases the difficulty of purification, but also is more soluble and easy to remain in the preparation, reducing the safety of medication
However, there is no report on the synthesis of creatine phosphate sodium by immobilized enzymes

Method used

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  • Preparation method for creatine phosphate sodium
  • Preparation method for creatine phosphate sodium
  • Preparation method for creatine phosphate sodium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, extract creatine kinase

[0029] The method for extracting creatine kinase comprises the steps of:

[0030] (1) Get 380g of frozen rabbit thigh muscle, cut into small pieces after removing connective tissue and nerves, then add 1100ml of KCl with a concentration of 0.01mol / L, then smash the rabbit meat with a meat grinder;

[0031] (2) Break the crushed rabbit meat with a colloid mill with a gap of about 2 μm for 5 to 10 minutes; then stir and extract at room temperature (25°C) for 30 minutes, then centrifuge at 8000r / min for 10 minutes at 0°C, Discard the precipitate, collect the supernatant, and record it as V1;

[0032] (3) Add finely ground NH to V1 4 Cl to NH 4 Cl concentration is 0.1mol / L, with 5mol / LNH 4 Adjust the pH to 9.0 with OH, then stir at room temperature (25°C) for 30min, add 1.5 times the volume of V1 of 95% cold ethanol, and stir at room temperature (25°C) for 2.5h, then at 0°C, 8000r / min Centrifuge for 10 minutes, discard the precip...

Embodiment 2

[0041] Embodiment 2, the influence of temperature on creatine kinase extraction

[0042] The effect of temperature on creatine kinase is manifested in two aspects: one is that with the increase of temperature, the solubility of creatine kinase in saline solution or aqueous solution increases; Reduced vitality.

[0043] Considering that in actual production, especially when extracting a large amount of enzymes, in order to reduce technical difficulty and production costs, the extraction process should be as simple as possible. According to the above theory, the effect of temperature on the extraction of creatine kinase was studied.

[0044]Design A, B, C three groups of experiments, A group of experiments is crushed in step (1) colloid mill, step (3) (4) volume fraction is 95% ethanol extraction is room temperature (25 ℃), other steps are 0 ℃ ; Group B experiments were controlled below 35°C during centrifugation, and the others were all controlled at room temperature (25°C); ...

Embodiment 3

[0046] Embodiment 3, the influence of pH on creatine kinase extraction

[0047] Enzymes (proteins) are amphoteric substances with an isoelectric point. If the pH of the solution matches the isoelectric point, the enzyme is likely to be precipitated in this case. Therefore, when extracting enzymes, the pH value of the solution should be selected within the range deviated from both sides of the isoelectric point; Extract with an acidic extract. According to the method of Example 1, extract under the conditions of pH 6, 7, 8, 9 and 10 respectively, then detect creatine kinase activity, the results are as follows figure 2 shown. Depend on figure 2 It can be seen that when the pH of the extract solution is 9, the activity per milliliter of enzyme solution is the highest, and a higher activity is maintained in the range of 7-9.

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Abstract

The invention discloses a preparation method for creatine phosphate sodium. The method comprises the specific steps that creatine jubase is fixed to carrier resin to obtain fixed creatine jubase, creatine and ATP are subjected to a reaction under the catalysis of the fixed creatine jubase at the temperature of 5-40 DEG C and the pH value of 7-11 on the basis that the mole ratio of the creatine to the ATP is 2:1-10:1 and the molar concentration of magnesium ions is 0.5-5 times that of the ATP till the concentration of creatine phosphate sodium is constant, and a reaction solution is collected; then the reaction solution is pretreated with the ethyl alcohol with the volume fraction being 40-70%, then, a pretreated creatine phosphate sodium mother solution is loaded, the creatine is eluted, then the creatine phosphate sodium is eluted, an eluent is collected, and crystallization and drying are performed to obtain the creatine phosphate sodium. The purity of the creatine phosphate sodium obtained through the preparation method is high and can reach 99.8%, the creatine phosphate sodium does not contain sulfate radicals or barium ions, the safety is high, and the medical standards are met.

Description

technical field [0001] The invention belongs to the field of chemistry, and in particular relates to a preparation method of creatine phosphate sodium. Background technique [0002] Phosphocreatine is an active substance in the human body. The substance is currently used clinically in the form of creatine phosphate sodium tetrahydrate, mainly for the treatment of cardiovascular diseases and metabolic diseases. The drug has a huge market, broad prospects, and a wide range of drug users. Up to now, the raw materials of creatine phosphate sodium include biological extraction method, chemical synthesis method, and free enzyme synthesis method, but there is no report on the synthesis of creatine phosphate sodium product by immobilized creatine kinase in China. The biological extraction method has the disadvantages of low yield and difficult quality control. The disadvantage of the chemical synthesis method is that the reaction process is intense and the reaction conditions are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N11/02
Inventor 陈真文廖凤霞程永刚李玉林彭力周玲杨红涛毛先兵
Owner XINAN PHARMA
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