Application of lorarexed or pharmaceutically acceptable salt thereof in preparation of medicine for treating or preventing herpes virus infection
A kind of herpes virus infection, loratrexed technology, applied in the field of biomedicine
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Embodiment 1
[0046] The application of targeting thymidylate synthase (TS) in screening anti-herpes virus drugs comprises the following steps:
[0047] Pemetrexed disodium is known as a thymidylate synthase inhibitor, and this inhibitor is used to verify whether it has anti-herpes virus effects:
[0048] 1. Experimental materials
[0049] 1.1 Cells and viruses
[0050] 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; 293T cells were preserved in our laboratory and purchased from American Type Culture Collection (ATCC).
[0051] Pemetrexed disodium was purchased from Biochempartner.
[0052] 1.2 Reag...
Embodiment 2
[0087] The application of targeting thymidylate synthase (TS) in screening anti-herpes virus drugs comprises the following steps:
[0088] Pemetrexed disodium is known as a thymidylate synthase inhibitor, and this inhibitor is used to verify whether it has anti-herpes virus effects:
[0089] 1. Experimental materials
[0090] 1.1 Cells and viruses
[0091] 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.
[0092] 1.2 Reagents
[0093] 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.
[0094] 1.3 Experimental Instruments
[0095] The multi-label microplate reader was purchased from PerkinElmer; the fluorescent quantitat...
Embodiment 3
[0118] The application of targeting thymidylate synthase (TS) in screening anti-herpes virus drugs comprises the following steps:
[0119] Pemetrexed disodium is known as a thymidylate synthase inhibitor, and this inhibitor is used to verify whether it has anti-herpes virus effects:
[0120] 1. Experimental materials
[0121] 1.1 Cells and viruses
[0122] 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.
[0123] 1.2 Reagents
[0124] DMEM medium and FBS were purchased from GIBCO; Alamarblue activity detection kit was purchased from Promega; crystal violet staining solution.
[0125] 1.3 Experimental Instruments
[0126] The multi-label microplate reader and the high-content cell analyzer were purchased fr...
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