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Application of thymidylate synthase as target in screening anti-herpes virus drugs

A thymidylate synthase and anti-herpes virus technology, applied in the field of biomedicine

Inactive Publication Date: 2016-02-24
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, drugs to treat or prevent herpes virus infection still need to be further developed
[0004] However, there is currently no application of thymidylate synthase as a target for screening antiherpes virus drugs

Method used

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  • Application of thymidylate synthase as target in screening anti-herpes virus drugs
  • Application of thymidylate synthase as target in screening anti-herpes virus drugs
  • Application of thymidylate synthase as target in screening anti-herpes virus drugs

Examples

Experimental program
Comparison scheme
Effect test

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] The iSLK.219 cell was donated by Professor Dr. DonGanem. The cell is a SLK cell containing the recombinant virus rKSHV.219 and the RTA expression system regulated by Doxycycline. In this cell, the RTA expression sequence was constructed into the pRetro-XTet-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 Reagent...

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 / mLTPA 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 GreenSupermix) was purchased from Bio-Rad Company.

[0094] 1.3 Experimental Instruments

[0095] The multi-marker microplate reader was purchased from PerkinElmer; the fluorescent quantitativ...

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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Abstract

The invention discloses application of thymidylate synthase as the target in screening anti-herpes virus drugs. thymidylate synthase can be used for screening drugs resisting Herpes simplex virus-1 (HSV-1), Herpes simplex virus-2 (HSV-2), Varicella zoster virus (VZV), EB virus (Epstein-Barr virus, EBV), Cytomegalovirus (CMV), human herpes virus type VI (HHV-6), human herpes virus type VII (HHV-7) and Kaposi's sarcoma-associated herpesvirus (KSHV) and other herpes viruses. The cell target thymidylate synthase involved in the invention can be used as a new anti-herpes virus target for drug development, and can be applied to treatment or prevention of herpes virus infection.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of targeting thymidylate synthase in screening anti-herpes virus drugs. Background technique [0002] Herpesviridae is a family of viruses with envelopes and double-stranded DNA genomes. Members of this family can widely infect animals and humans and induce corresponding diseases. There are currently eight herpes viruses that can infect humans, including: Herpes simplex virus-1 (HSV-1), Herpes simplex virus-2 (HSV-2), Varicella-Zoster Herpes virus (Varicellazostervirus, VZV), Epstein-Barrvirus (EBV), cytomegalovirus (Cytomegalovirus, CMV), human herpesvirus type VI (HHV-6), human herpesvirus type VII (HHV-7) and card Bordeaux herpes virus (Kaposi's sarcoma-associated herpesvirus, KSHV). According to the differences in genome sequence, structure and physical and chemical properties, Herpesviridae can be divided into three subfamilies: α-Herpesvirinae, β-Herpesvirinae ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50A61K45/00A61K45/06A61K31/519A61K31/4709A61P31/22
CPCA61K31/4709A61K31/519A61K45/00A61K45/06G01N33/5014
Inventor 陈绪林陈军刚
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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