A pharmaceutical composition containing urate oxidase and its preparation method
A uric acid oxidase and composition technology, which is applied in the directions of medical preparations containing active ingredients, drug combinations, and non-active ingredients medical preparations, etc., can solve inflammatory reactions, recurrent gout attacks, and the inability of PEGylated uric acid oxidase to enter. And other issues
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Embodiment 1
[0084] Example 1 UOX derived from Candida
[0085] (1) Preparation of PC-UOX composition
[0086] Add 30 mL of boric acid buffer (50 mM boric acid, 50 mM potassium chloride) containing 1 mg / mL of Candida-derived UOX to a 100 mL reagent bottle at pH 8.5, and place at 4°C.
[0087] Refined lecithin (0.012 g) with a purity greater than 98% was dissolved in 6 mL of isopropanol to prepare a lecithin solution. The solution temperature was 4°C. 6 mL of this lecithin solution was added dropwise to the aforementioned UOX solution at a rate of 1 mL / min, and the solution obtained by stirring at 50 rpm for 4 hours was used as a reaction solution.
[0088] The reaction solution was added to a G-25 desalting column equilibrated with carbonate buffer (20 mM pH10.5 sodium carbonate-sodium bicarbonate buffer, 100 mM sodium chloride, 0.1% Tween-20) for desalting and buffer exchange. The obtained solution was added to an anion exchange column equilibrated with carbonate buffer (20 mM pH10.5 s...
Embodiment 2
[0104] The UOX of embodiment 2 Aspergillus flavus origin
[0105] (1) Preparation example of PC-UOX composition
[0106] Add 30 mL of pH 8.0 boric acid buffer (50 mM boric acid, 50 mM potassium chloride) containing 1 mg / mL of Aspergillus flavus-derived UOX to a 100 mL reagent bottle, and place it at 4 °C.
[0107] Refined lecithin (0.012 g) with a purity greater than 98% was dissolved in 6 mL of isopropanol. Prepare lecithin derivative solution. The solution temperature was 4°C. 6 mL of the purified lecithin residue solution was added dropwise to the aforementioned UOX solution at a rate of 1 mL / min, and the solution obtained by stirring at 50 rpm for 4 hours was used as a reaction solution.
[0108] The reaction solution was added to a G-25 desalting column equilibrated with carbonate buffer (20 mM pH10 sodium carbonate-sodium bicarbonate buffer, 100 mM sodium chloride, 0.1% Tween-20) for desalting and buffer exchange. The obtained solution was added to an anion exchange ...
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