Pharmaceutical application of combination of salvianolic acid A, salvianolic acid B and salvianolic acid C for resisting 2019-nCov virus and medicine
A 2019-ncov, salvianolic acid technology, applied in the field of antiviral drug preparation
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Embodiment 1
[0025] Embodiment 1, SPR method detects the binding ability of salvianolic acid A, salvianolic acid B and salvianolic acid C and ACE2 protein and S protein
[0026] Experimental materials: ACE2 protein (purchased from Beijing Yiqiao), salvianolic acid A (purchased from Baoji Chenguang), salvianolic acid B (purchased from Shanghai Anpu) and salvianolic acid C (purchased from Baoji Chenguang), SPR, COOH Chips and corresponding activation reagents (purchased from Nicoya, Canada).
[0027] Preparation method: install a carboxyl chip in the Open SPR instrument, and pump the running buffer at the maximum flow rate (150 μL / min) to make it full of the detection cell. After reaching the signal baseline, inject 80% isopropanol to degas and clean the chip surface with 10 mM HCl. After reaching the signal baseline again, adjust the flow rate to 20 μL / min. Immediately mix an equal amount of coupling reagent (EDC / NHS) and inject it into the injection valve, activate the chip for 5 minutes...
Embodiment 2
[0030] Example 2. Salvianolic acid A, salvianolic acid B and salvianolic acid C inhibit 2019-nCov pseudovirus from infecting cells with high expression of ACE2.
[0031] ACE2 high-expressing cells were seeded in 96-well plates, and each well was supplemented with medium to 50 μL. 37°C, 5% CO 2 Incubate in the incubator for 2 hours to adhere to the wall. Add 20 μM salvianolic acid A, salvianolic acid B or salvianolic acid C prepared in culture medium respectively, add 5uL 2019-CoV-2 Spike pseudovirus to each well for 4 hours, make up to 100 μL culture volume, continue infection for 6-8 hours, replace Make 200 μL of new complete medium and continue culturing at 37°C for 48h. The Luciferase Assay System kit detects the luminescence value of Luciferase.
[0032] see image 3 , after administration, the luminous intensity of Luciferase decreased compared with the control group. Chloroquine (20 μM) was used as an antiviral positive control. From image 3 It can be seen from t...
Embodiment 3
[0033] Example 3 Combined administration of salvianolic acid A, salvianolic acid B and salvianolic acid C inhibits 2019-nCov pseudovirus from infecting cells with high expression of ACE2.
[0034] ACE2 high-expressing cells were seeded in 96-well plates, and each well was supplemented with medium to 50 μL. 37°C, 5% CO 2 Incubate in the incubator for 2 hours to adhere to the wall. 10 μM salvianolic acid B or 2.5 μM salvianolic acid A, 5 μM salvianolic acid B and 2.5 μM salvianolic acid C were administered alone, and incubated at 37°C for 2 hours. Add 5uL 2019-CoV-2 Spike pseudovirus to each well to infect for 4 hours, make up to 100 μL culture volume, continue infection for 6-8 hours, replace with 200 μL new complete medium, and continue to culture at 37°C for 48 hours. The Luciferase AssaySystem kit detects the luminescence value of Luciferase.
[0035] see Figure 4 , after co-administration, the luminous intensity of Luciferase decreased compared with the control group. ...
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