Controlled-release preparation using live erythrocyte to load betamethasone sodium phosphate and preparation method and application thereof
A technology of sodium metasone phosphate and slow-release preparations, which is applied to medical preparations with no active ingredients, medical preparations containing active ingredients, anti-inflammatory agents, etc., can solve the problems of short half-life, large side effects, side effects, etc., and achieve a solution Side effects, obvious sustained release effect, and the effect of solving the rapid decline of blood drug concentration
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Embodiment 1
[0041] Example 1 Preparation of Packed Red Blood Cells
[0042] Male SD rats with a body weight of about 300 g were taken, and blood was taken from the retro-orbital venous plexus with a capillary tube with a diameter of 0.5 mm, and the blood volume was 0.2-0.3 ml each time. Centrifuge at 3800r / min at 4°C for 5min, discard the supernatant, remove the plasma and white blood cell layer, then add 1ml of pre-cooled PBS to wash 1-2 times until the supernatant is nearly colorless, and the lower layer is packed red blood cells.
Embodiment 2
[0043] The preparation of embodiment 2 drug-loaded erythrocytes
[0044] (1) Prepare a hypotonic betamethasone sodium phosphate medicinal solution with a mass fraction of 0.55% sodium chloride and a concentration of 4 mg / ml betamethasone sodium phosphate, and the solvent is water.
[0045] (2) Take 0.1ml of packed red blood cells prepared in Example 1 and add them to 1ml of hypotonic betamethasone sodium phosphate medicinal solution, mix gently, and place at 4°C for 30min to obtain live red blood cells-betamethasone sodium phosphate medicinal solution .
[0046] (3) Add 0.1ml hypertonic nutrient solution (mass fraction is 5.5% sodium chloride, 5mg / ml sodium pyruvate, 10mg / ml glucose, solvent is water) in 1ml live erythrocyte-betamethasone sodium phosphate medicinal solution, Mix gently and incubate at 37°C for 30min.
[0047] (4) Centrifuge the mixture obtained in step (3) at 4°C, 2000r / min, 5min / time, remove the supernatant and wash with pre-cooled PBS for 3 times under the...
Embodiment 3
[0048] The mensuration of embodiment 3 drug loads
[0049]Collect the supernatant of each step in Example 2, draw 0.2ml, add 1ml methanol and mix, centrifuge at 12000r / min for 30min, take the supernatant and filter it with a 0.22 μm disposable filter membrane, and inject it into a high-performance liquid chromatograph. The content of free betamethasone sodium phosphate in the supernatant was determined.
[0050] Drug loading=the total amount of betamethasone sodium phosphate added-the content of free betamethasone sodium phosphate in the supernatant.
[0051] Wherein the measuring condition of high performance liquid chromatography is:
[0052] Column: Agilent XB-C18 (5μm, 4.6×250nm); detector: AgilentTechnologics UV detector; mobile phase: methanol-0.05mol / l potassium dihydrogen phosphate (1:1); flow rate: 1ml / min; column temperature: 40°C; Detection wavelength: 254nm.
[0053] The final drug loading measurement result was 2.5 mg / ml (ml refers to the volume of packed red...
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