Medicine for repairing and treating viscera injury
A drug and organ technology, applied in the field of biopharmaceuticals, can solve the problems that have not yet been discovered, drugs that do not use PEG-rhG-CSF to damage organs, increase half-life, etc., achieve molecular weight increase, half-life increase, and reduce antigenicity Effect
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Embodiment 1
[0051] Example 1. Contains PEG 20000 -drugs for rhG-CSF
[0052] (1) Preparation of PEG 20000 -rhG-CSF:
[0053] ①Concentrate the GC solution by ultrafiltration, replace the buffer system with 0.1M phosphate buffer (pH8.5), and obtain 1000ml of GC solution with a concentration of 5mg / ml.
[0054] ②Add 1.66g of DMMAn, keep the temperature of the reaction system at 0°C in an ice bath, and maintain the reaction for 30min.
[0055] ③Add 52.6g mPEG to the reaction system 20000 -acetaldehyde, stir to dissolve, heat up to 37°C, add 0.16g NaBH 3 CN, reacted for 1 hour. The above reaction mixture was adjusted to pH 6.0 with 0.1M HCl at 37°C for 30 minutes.
[0056] ④ Load the reaction product and elute with a linear gradient of 10mM sodium acetate buffer (pH4.0) and 0-0.45M sodium chloride on SPSepharose Big Beads ion column equilibrated with 10mM sodium acetate buffer (pH4.0) 6 column volumes, the main peak is collected as mPEG 20000 -rhG-CSF. Desalt through Superdex 75 colum...
Embodiment 2
[0063] Example 2. Contains PEG 30000 -drugs for rhG-CSF
[0064] (1) Preparation of PEG 30000 -rhG-CSF:
[0065] ①Concentrate the GC solution by ultrafiltration, replace the buffer system with 0.1M phosphate buffer (pH8.5), and obtain 1000ml of GC solution with a concentration of 5mg / ml.
[0066] ② Add 1.83g DMMAn, keep the temperature of the reaction system at 0°C in an ice bath, and keep the reaction for 30min.
[0067] ③Add 86.8gmPEG to the reaction system 330000 -acetaldehyde, stir to dissolve, heat up to 39°C, add 0.176g NaBH 3 CN, reacted for 1.5 hours. The above reaction mixture was adjusted to pH 6.2 with 0.1M HCl at 35°C for 40 minutes.
[0068] ④ Load the reaction product, and wash with 10mM sodium acetate buffer (pH4.0) and 0-0.6M sodium chloride linear gradient on the SP Sepharose Big Beads ion column equilibrated with 10mM sodium acetate buffer (pH4.0). Remove 10 column volumes, collect the main peak as mPEG 30000 -rhG-CSF. Go through the Superdex 75 colu...
Embodiment 3
[0077] PEG prepared by the method described in Example 1 20000 -Effects of rhG-CSF preparations on cardiac hemodynamics and myocardial oxygen consumption in anesthetized dogs: PEG 20000 -rhG-CSF preparations have no direct and immediate effect on cardiac hemodynamics and myocardial oxygen consumption in anesthetized mongrel dogs at two doses of 15 and 30 μg / kg, indicating that PEG-rhG-CSF treats coronary artery disease. Heart disease, reducing the size of myocardial infarction, and improving heart function are not achieved by directly affecting cardiac hemodynamics and myocardial oxygen consumption, but by promoting the release of autologous bone marrow stem cells and targeted transplantation In the heart, it can promote the regeneration of myocardial cells and small myocardial blood vessels, repair the disease and play a role in the treatment of myocardial infarction. Its mechanism of action is the result of autologous bone marrow stem cell transplantation, rather than the d...
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