In-vivo and in-vitro anti-RSV (Respiratory Syndrome Virus) licorice root effective part as well as preparation method and application thereof
A technology of effective parts and licorice, applied in the field of medicine, can solve problems such as teratogenic effects and controversial effects, and achieve the effects of small side effects, easy operation and simple process.
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[0078] The preparation method of licorice effective parts for anti-RSV in vivo, including the following steps:
[0079] (1) Plus water, soak, heat reflux extraction, concentrate, glycycetic water extract;
[0080] (2) The licorice water extract is extracted from petroleum ether, dichloromethane, ethyl acetate, and n-butyl alcohol, retain the n-butanol extract, and produce a licorice effective site.
[0081] The specific preparation method of licorice effective parts, including the following steps:
[0082] (1) Take 200 g of licorice herbs, washed, add 12 times licorice with weight of water, soak 30 min, heat reflux extraction of 90 min, a total of twice, then centrifuge after the extraction liquid, concentrate, glyccetic water extraction liquid;
[0083] (2) Add petroleum ether and petroleum ether in the glycylder water extract, the volume ratio of the petroleum ether is 1: 1, extract 2-3 times with petroleum ether, and the petroleum ether layer is colorless, separated, and retain...
Embodiment 1
[0089] Example 1 in vitro anti-RSV study of licorice effective parts
[0090] 1.1 sample preparation:
[0091] Preparation of the test solution: DMEM cell culture containing 2% FBS was prepared into a 6 mg / ml solution.
[0092] Preparation of the Tollic Solution: DMEM cell culture medium containing 1% DMSO and 2% FBS separates glycyrrhizic acid, licoriceic acid, glycyrrhizalic acid single ammonium salt, and ribavirin, 1mm glycyrridic acid solution, licorice acid Solution, mono ammonium salt solution, Libelin solution.
[0093] 1.2 Cell culture and recovery
[0094] The preserved cells were removed from the liquid nitrogen tank, rapidly melted at 37 ° C. The cell suspension was transferred to a sterile centrifuge tube in a biosafety cabinet, centrifuged (1000R, 3 min), abandoned to the supernatant, added a new cell culture medium, blowing the cell, transfer the suspension to the cell culture dish Co 2 Culture Culture Culture (Culture Conditions 37 ° C, 5% CO 2 Relative humidity 7...
Embodiment 2
[0106] Example 2 Research on the action phase of the Time experiment to investigate the role of the RSV replication cycle
[0107] In order to study the specific phase of the n-butanol part in the RSV replication cycle, the overtime experiment was carried out. Hep2 cells were 8 × 10 in 24 hours before the experiment. 4 The cells were laid into a 24-well plate per well, and the cells were infected with a viral of MOI 3 at 4 ° C for 2 hours at 0 h, followed by washing cells three times with cold PBS and the new culture medium was cultured in a cell incubator. The n-butanol portion (0.3 mg / ml) and the positive drug levein (100 μm) were added at -2, 0, 2, 4, 6, 8, 12, 16h, respectively, and the supernatant was collected at 20 h and The liquid viral virus is measured in accordance with the measurement method of 1.4 in Example 1.
[0108] Test results Figure 6 Sum Figure 7 . Reference Figure 6, Positive control of drugs, Rashilin has a better inhibitory effect on RSV at 6 to 10 h, mai...
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