Application of Tilorone in prevention/treatment of African swine fever virus infection
An African swine fever virus and application technology, applied in the field of medicine, can solve the problems of research and disclosure without inhibitory/therapeutic effects, and achieve the effects of excellent oral bioavailability, high selectivity coefficient, and multiple modes of administration
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Embodiment 1
[0035] Study on Inhibition of African Swine Fever Virus Infection Stage by Tilorone Hydrochloride
[0036] Take 2×10 5 Cells / well were inoculated on lysine-coated 96-well plates and incubated in a cell culture incubator for 24 hours. Infect with 1MOI, and add Tyrolone (such as figure 2 indicated), at a concentration of 10 μM. At the same time, in order to exclude the direct toxic effect of the drug on the virus, the virus and Tyrolone were mixed evenly, and the final concentration of the drug was 10 μM. After incubation at 37°C for 1 hour, the mixture was diluted 100 times, and the cells were infected. At this time, the infection dose was 1 MOI. . Collect samples within 24 hours to detect virus titer HAD 50 and genome copy number. see results image 3 . It shows that Tilorone hydrochloride has obvious inhibition to African swine fever virus.
Embodiment 2
[0038] In vitro activity and cytotoxicity experiments of Tilorone hydrochloride inhibiting the replication of African swine fever virus
[0039] By measuring CC 50 To evaluate the toxicity of Tilorone hydrochloride to cells: with 4×10 5 Porcine alveolar macrophages (porcine alveolar macrophages, PAM) were inoculated in a 96-well plate at a density of one per well and incubated for 24 hours.
[0040] Drug configuration and dilution method are as follows: Tirolone hydrochloride is dissolved in DMSO, is configured into a 1mM solution, and then diluted to 0.2mM with 1640 medium containing 10% FBS and 2% double-antibody SP (penicillin and streptomycin) as the initial concentration , and then sequentially carried out 2-fold dilution to form 8 gradients, and each gradient was repeated for 3 replicate wells.
[0041] After the cells were cultured for 24 hours, discard the cell culture supernatant, add 100 μL / well of the diluted drug, and place it in the cell culture incubator for 72...
Embodiment 3
[0047] Pharmacokinetics of Tilorone Hydrochloride in Miniature Bama Pigs
[0048] Three healthy male and three female Bama pigs were given Tilorone hydrochloride by intravenous injection and gavage respectively, and the blood drug concentration of the test compound Tilorone hydrochloride in plasma was quantitatively determined by liquid chromatography-tandem mass spectrometry, and the pharmacokinetic parameters were calculated to investigate the Pharmacokinetic characteristics of compounds in miniature Bama pigs.
[0049] 3.1 Experimental materials:
[0050] animal species Pig animal strain mini parmesan pig animal age 0.5-1years Animal sex and number 6, half male / female animal weight 11.0-14.0kg animal source Jiangsu Langyue Biotechnology Co., Ltd. Certificate No. 3260374713
[0051] 3.2 Animal feeding:
[0052] Animals were kept in single cages in the Experimental Animal Center of Suzhou Laio Biotechnology Co., Ltd. ...
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