Application of raltitrexed or pharmaceutically acceptable salt thereof in preparation of medicine for treating or preventing herpes virus infection
A technology of herpes virus infection and raltitrexed, which is applied in the field of biomedicine to achieve the effect of good anti-herpes virus activity
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Embodiment 1
[0033] Detection of the activity of raltitrexed in inhibiting KSHV lytic phase replication
[0034] 1. Experimental materials
[0035] 1.1 Cells and viruses
[0036]iSLK.219 cells were donated by Professor Dr. Don Ganem. The cells are SLK cells containing recombinant virus rKSHV.219 and doxycycline (Doxycycline) regulated RTA expression system. In this cell, the RTA expression sequence was constructed into the pRetro-X Tet-ON inducible expression system (Clontech, Mountainview, CA), and the expression of this system was regulated by doxycycline. In the presence of doxycycline, RTA expression can occur; Vero cells were preserved in our laboratory and purchased from American Type Culture Collection (ATCC).
[0037] Raltitrexed was purchased from Biochempartner.
[0038] 1.2 Reagents
[0039] DMEM medium, 1640 medium and FBS were purchased from GIBCO; Alamarblue activity detection kit was purchased from Promega; SYBR mixture (iTaq TM Universal Green Supermix) was purchased...
Embodiment 2
[0065] Embodiment 2: the detection of raltitrexed anti-HSV-1 and HSV-2 activity
[0066] 1. Experimental materials
[0067] 1.1 Cells and viruses
[0068] Vero cells were preserved in our laboratory and purchased from the American Type Culture Collection (ATCC). HSV-1 is an F strain, which is preserved by the Wuhan Institute of Virology, Chinese Academy of Sciences. HSV-2 is a G strain, which is preserved by the Wuhan Institute of Virology, Chinese Academy of Sciences.
[0069] 1.2 Reagents
[0070] DMEM medium and FBS were purchased from GIBCO; Alamarblue activity detection kit was purchased from Promega.
[0071] 1.3 Experimental Instruments
[0072] The multi-label microplate reader and the high-content cell analyzer were purchased from PerkinElmer; the cell incubator was a product of Thermo.
[0073] 2. Experimental methods and results
[0074] 2.1 Cytotoxicity detection of raltitrexed on Vero cells
[0075] (1) Divide Vero cells into 8×10 3 Each cell / well was see...
Embodiment 3
[0094] Embodiment 3: Raltitrexed inhibits the detection of EBV lytic phase replication
[0095] 1. Experimental materials
[0096] 1.1 Cells and viruses
[0097] The B95-8 cell was donated by Professor Sun Xiaoping of the School of Basic Medicine of Wuhan University. The cell contains the complete sequence of the EBV virus genome. Combined induction with 20ng / mL TPA can make the virus enter the lytic phase and replicate.
[0098] 1.2 Reagents
[0099] 1640 medium and FBS were purchased from GIBCO; Alamarblue activity detection kit was purchased from Promega; SYBR mixture (iTaq TM Universal Green Supermix) was purchased from Bio-Rad Company.
[0100] 1.3 Experimental Instruments
[0101] The multi-marker microplate reader was purchased from PerkinElmer; the fluorescent quantitative PCR instrument (Bio-RadCFX96Touch TM Real-Time PCR detection system) was purchased from Bio-Rad Company; Type refrigerated centrifuge and cell culture box were purchased from Thermo Company...
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