Application of antibacterial peptide BF-30 in preparation of medicines for treating or preventing thrombotic diseases
A technology of BF-30 and thrombotic diseases, applied in the field of biomedicine, can solve problems such as no public reports, and achieve the effect of prolonging APTT time and inhibiting platelet aggregation
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Embodiment 1
[0030] Example 1: In vitro anti-platelet aggregation activity of BF-30
[0031] 1) Purchase human apheresis platelets from Jiangsu Provincial Blood Center, dilute in half with modified Tyrode's solution to obtain platelet-rich plasma, and then centrifuge at 3000 rpm for 10 minutes to obtain platelet-poor plasma.
[0032] 2) Take 300 μl of platelet-poor plasma and place it in the test area of the platelet aggregometer for zero adjustment, then draw 270 μl of platelet-rich plasma and place it in the preheating tank, add BF-30 with different concentration gradients (10, 15, 25, 35, 50 μM) And test beads, preheated at 37°C for 5 minutes, placed in the test area, and added 10 μl of inducer thrombin solution (final concentration 0.26 U / ml). Determine the maximum aggregation rate of platelets within 5 min. Wherein, the blank control is normal saline, and the positive control is tirofiban. The platelet aggregation inhibition rate was calculated by the following formula: [(X-Y) / X]×...
Embodiment 2
[0037] Example 2: Effect of BF-30 on APTT and PT
[0038]1) Take rabbit carotid artery blood, place it in a 15ml centrifuge tube containing 1ml of 3.8% sodium citrate (the final volume of each tube is about 10ml, and ensure that the volume ratio of rabbit blood and anticoagulant is 9:1), Centrifuge at 3000rpm for 10min to obtain platelet-poor plasma.
[0039] 2) Add 20 μl of platelet-poor plasma and 5 μl of normal saline or BF-30 (50 μM) to the test cup, incubate at 37°C for 3 minutes, then add 25 μl of APTT reagent, continue to incubate for 3 minutes, place it in the test area, add 25 μl of 37°C CaCl2 solution, read the APTT time on the hemagglutination meter.
[0040] 3) Add 20 μl of platelet-poor plasma and 5 μl of normal saline or BF-30 (50 μM) into the test cup, incubate at 37°C for 3 minutes, then place it in the test area, add 50 μl of PT reagent at 37°C, and read the hemagglutination meter PT time.
[0041] 4) Through APTT and PT experiments, BF-30 can significantly...
Embodiment 3
[0045] Example 3: Effect of BF-30 on ADP-induced acute pulmonary thrombosis in mice
[0046] 1) Take 50 ICR mice, weighing 18-22g, divide them into 5 groups, 10 in each group, inject normal saline, 50mg / kg aspirin into the tail vein respectively, and dilute with normal saline to 6.3, 7.2, 9mg / kg BF -30. Wherein the negative control is physiological saline, and the dosage is 5ml / kg.
[0047] 2) After 15 minutes of administration (after 30 minutes of administration in the aspirin group), 300 mg / kg ADP solution was injected into the tail vein to form acute pulmonary thromboembolism, and the number of dead or hemiplegic mice in each group within 15 minutes after ADP injection was recorded. The mortality rate of each group was calculated and the protection rate of acute pulmonary thromboembolism of antithrombotic active substances was calculated by the following formula: [1-(X / Y)]×100%. X represents the number of dead mice in each group; Y represents the total number of mice in e...
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