Preparation and application of uric acid lowering oral medicine

A technology for reducing uric acid and uricase, which is applied in the field of preparation of oral uric acid-lowering drugs, can solve the problems of insufficient support and failure to explain the source of uricase, and achieve high safety effects

Inactive Publication Date: 2017-06-30
YUNNAN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

4 in Table 1 is to make uricase into an enteric-coated preparation orally to reduce serum uric acid, but the description example of the invention cannot fully support the existence of the curative effect, such as not explaining the source of uricase, and how to use uricase in rats orally Avoid the destructive effect of digestive enzymes, etc.

Method used

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  • Preparation and application of uric acid lowering oral medicine
  • Preparation and application of uric acid lowering oral medicine
  • Preparation and application of uric acid lowering oral medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation and activity determination of uricase.

[0042] According to the sequence (XM_002377830.1) of the mRNA of Aspergillus flavus uricase, the full length of the coding sequence was chemically synthesized, and then connected to the pET28a vector (with Karabacteria sp. protein resistance), Uricase-pET28a was obtained, and the correct encoding of uricase protein (see figure 1 ) vector to transfect Escherichia coli DH5a to obtain a large number of vectors, the DH5a bacteria were cultured in large quantities and the bacteria were recovered, and the Uricase-pET28a vector was extracted after the bacteria were lysed. The Uricase-pET28a vector was purified and introduced into BL21(DE3) Escherichia coli, and positive bacteria were obtained by karimycin resistance screening. Then use the LB medium containing karimycin resistance (50 μg / ml) to cultivate Uricase-pET28a-BL21 (DE3) bacteria in large quantities on a shaker (160 rpm) at 37 ° C. When the bacteria propa...

Embodiment 2

[0044] Example 2 Preparation of Bacillus subtilis macromolecular extract with uricase activity.

[0045] Inoculate live medical or edible Bacillus subtilis into 1L sterilized medium (containing 3.0g beef extract, 5.0g peptone per liter, pH 6.8±0.2), shake at 37°C (160rpm) until the turbidity OD600 reaches 2 Above, then centrifuge for 10 minutes to recover the bacteria (5000rpm, 4°C), and the macromolecular extraction sample is called the pure Bacillus subtilis group from the bacteria. Another bottle of bacteria was cultured in parallel, the method was the same as before, but 10 g of uric acid was added during the culture, and the bacteria were also recovered. The macromolecular extraction sample came from the uric acid-induced Bacillus subtilis group of the bacteria. The recovered bacteria were resuspended with pH 7.40 phosphate buffer, crushed with an ultrasonic crusher, and centrifuged to obtain supernatant (to remove particles such as cell nuclei). Put the supernatant in a...

Embodiment 3

[0050] Example 3 Preparation of yeast macromolecule extract with uricase activity.

[0051] Inoculate edible beer yeast live bacteria (yeast powder) into 1L sterilized medium (each liter of culture medium contains 3.0g of yeast extract, 3.0g of malt extract, 10.0g of glucose, 5.0g of peptone, pH6.2±0.2) , Shake at 37°C (160rpm) until the turbidity OD600 reaches above 2, then centrifuge for 10 minutes to recover the yeast (5000rpm, 4°C), the macromolecule extraction sample is called the pure yeast group. Another bottle of yeast was cultured in parallel, the method was the same as before, but 10 g of uric acid was added during the cultivation, and the yeast was also recovered, and the macromolecule extraction sample came from the uric acid-induced yeast group of the bacteria. The recovered bacteria were resuspended with pH 7.40 phosphate buffer, crushed with a high-pressure crusher, and then centrifuged to obtain supernatant (to remove particles such as cell nuclei). Put the su...

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Abstract

The invention discloses a uric acid lowering oral enteric-coated preparation. The uric acid lowering oral enteric-coated preparation is characterized by containing uricase, alkalescent additives and uricase protecting agent, wherein the uricase is recombinant uricase, macromolecular bacteria extract containing uricase or macromolecular fungus extract containing uricase; the alkalescent additives are an alkalescent mixture prepared from sodium bicarbonate, sodium phosphates, sodium acetate and other medical salts of organic weak acids and inorganic weak acids; the uricase protecting agent is edible protein powder, which is rich in protein content and free from obvious proteinase activity or even denatured. The uric acid lowering oral enteric-coated preparation degrades uric acid in intestines to reduce the serum uric acid level and takes effects without reliance on absorption, thereby avoiding liver and kidney toxicity, adverse cardiovascular response or other post-absorption adverse response led by traditional uric acid lowering medicines and further being applicable to prevention of hypeluricemia and gout.

Description

technical field [0001] The invention belongs to the technical field of pharmacy, and in particular relates to a preparation method and application of an oral uric acid-lowering drug. Background technique [0002] Hyperuricemia and gout have become common and frequently-occurring diseases that endanger human health. They are more common in men than women, and show a certain age correlation. The mechanism of hyperuricemia and gout is currently clear, mainly due to the disorder of purine metabolism in the body leading to excessive uric acid content. As an insoluble substance, uric acid, the final product of purine metabolism, is easily deposited in the kidneys, terminal joints, and auricles, resulting in kidney damage and various inflammatory reactions, which seriously affect the quality of life and even endanger life. In order to prevent and control the occurrence of gout, it is generally recommended that the uric acid content in serum should not exceed 420 μM (70 μg / ml). ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/44A61K47/42A61P19/06
CPCA61K9/0053A61K38/44A61K47/42C12Y107/03003
Inventor 段为钢云宇殷华吕小满李月段金连白雯
Owner YUNNAN UNIV OF TRADITIONAL CHINESE MEDICINE
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