Application of Brassica napus-Isatis g monomer addition line in inhibiting novel coronavirus sars-cov-2
A technology of Brassica napus and coronavirus, applied in the fields of application, antiviral agents, respiratory diseases, etc., can solve the problems of unclear pathogenesis and virus-host interaction molecular mechanism, no virus treatment, etc., and achieve no resistance Drug properties, low cost, large output effect
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Embodiment 1
[0050] Preparation of Brassica napus-Isatis indigo line extract:
[0051] The whole plants of different additional lines of Brassica napus-Isatis indica planted in the field were harvested, dried naturally in the shade or at low temperature, crushed, and passed through an 80-mesh sieve, and the collected powder was used for the preparation of crude extraction. The specific extraction steps are as follows:
[0052] 1) Add 100g of different additive powders and 1000mL of methanol into a 2000mL Erlenmeyer flask. Ultrasonic treatment at 55°C for 40-60 minutes, after filtration, collect the supernatant and filter residue respectively;
[0053] 2) Add 1000mL methanol to the filter residue in step 1 to continue ultrasonic treatment, cycle 5-6 times according to step 1, and combine all the supernatants obtained;
[0054] 3) Evaporate methanol from the supernatant in step 2 in a rotary evaporator at 50°C;
[0055] 4) Dissolve and evaporate the crude extract of methanol with single di...
Embodiment 2
[0060] Effects of extracts of different additional lines of Brassica napus-Isatis indica on the proliferation of SARS-CoV-2 virus at the cell level in vitro
[0061] 1. Cell culture
[0062] The frozen and revived Caco-2 cells were subcultured twice, and expanded with DMEM medium containing 10% fetal bovine serum and double antibodies (penicillin 100 U / ml, streptomycin 100 ug / ml).
[0063] 2. The cells were treated with the extract of Brassica napus-Isatis indigo line and infected with the new coronavirus WBP-1
[0064] 1) The Caco-2 cells in good growth state were digested and passaged, and the cell density was adjusted to 1×10 with cell culture medium. 5 / ml, 1ml per well was inoculated in a 12-well plate.
[0065] 2) When the cell growth monolayer density reaches 80%, add the extracts of different additional lines of Brassica napus-Isatis indigo to the Caco-2 cells in the 12-well plate, the extract concentration is 200mg / mL, and the working concentration is 400μg / mL mL. ...
Embodiment 3
[0078] Inhibitory effect of Brassica napus-Isatis indigo G monomer addition line on the proliferation of SARS-CoV-2 virus at the cell level in vitro:
[0079] According to the results of Example 2, it can be known that the G monomer addition system extract has a significant inhibitory effect on the new coronavirus when the working concentration is 4mg / mL, and Example 3 further confirms the anti-new coronavirus effect and the The inhibitory effect of different concentrations on the proliferation of the new coronavirus.
[0080] 1. Toxic effect of G monomer extract on cells
[0081] 1) Take well-grown Caco-2 cells for digestion and passage, and use cell growth medium (DMEM medium + 10% fetal bovine serum + double antibody) to adjust the cell density to 2 × 10 6 / ml, inoculate 96-well plate, inoculate 100μl per well;
[0082] 2) Add 100 μl G monomer extracts of different concentrations prepared with culture medium (DMEM medium + 10% serum + double antibody) to each well, and mi...
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