Discovery And Mechanism Study Of Small Molecule Active Compounds That Regulating The Replication Of HSV-2

Herpes simplex virus type 2(HSV-2)is one of the most infectious sexually transmitted viruses,which has infected more than 400 million people worldwide so far.Due to the existence of asymptomatic infections,the actual number may be much higher.There is currently no effective vaccine or prophylaxis can completely immune or prevent the infection of HSV-2.The treatment of viral infection mainly depends on nucleoside antiviral drugs.There are two states of HSV-2 infection,namely lytic infection and latent infection.Although nucleoside drugs can alleviate the symptoms caused by lytic infection,they cannot eliminate the virus or inhibit the latency of it.At the same time,the emergence of resistant strains limits the use of nucleoside drugs.Therefore,the infection of HSV-2 cannot be completely cured at present.And because of the presence of latent infection,infected patients will periodically show symptoms of the disease.So,it is urgent to find antiviral drugs that acting on other targets and treatment options specific to latent infection.Ion channels are a class of special membrane proteins that selectively mediate the passage of ions through cell membranes,and participate in important physiological and pathological processes such as signal transduction,cell cycle regulation,and apoptosis.Studies have shown that compounds that target ion channels can resist viral invasion by blocking the life cycle of the virus.In this article,we screened the compound library of ion channels regulators and found that voltage-gated calcium channel blockers including lercanidipine,benidipine and lomerizine could inhibit the infection of HSV-2 in a concentration-dependent manner,the half inhibitory concentration(IC50)of which were 10.97,11.52,and 16.92?M,respectively.In the investigation of the stage of action of benidipine,we found that benidipine does not affect the entry and nuclear transport of HSV-2,and its antiviral effect is mainly reflected in the late stage of viral infection.In addition,T-type calcium channel-specific blocker Mibediel or knockdown of Cav3.1 can also affect the expression of HSV-2 genes and proteins.Further mechanistic studies have shown that benidipine affected the expression of viral genes and proteins by inhibiting the nuclear export of HDAC3 and reducing the histone acetylation level.In addition,based on the previous discovery of the role of resveratrol in activating latent HIV,we tested resveratrol and its derivatives to identify compounds that affect HSV-2 infection.We found that resveratrol and triacetyl resveratrol accelerated replication of HSV-2 and increased the release of progeny virion in a concentration dependent manner,and both of them had lower cytotoxicity in the range of working concentrations.A time-of-addition study suggested that resveratrol worked primarily in the early stage of viral infection,while the promotion effect on HSV-2 of the late stage administration was not obvious.Further mechanistic studies have shown that resveratrol regulated HSV-2 infection by increasing histone acetylation and activating NF-?B.In addition,inhibition of CDK9 activity restrained the promoting effect of resveratrol on HSV-2 infection.In conclusion,our study provides an experimental basis for calcium channel inhibitors as drugs against HSV-2 infection and voltage-gated calcium channels as antiviral targets.Meanwhile,the discovery of the role and mechanism of resveratrol in promoting HSV-2 infection also provide new ideas for further understanding and studying the regulatory mechanism of HSV gene expression and the reactivation of the latent virus.