The Screening And Antiviral Activity Of Tomato Chlorotic Virus Minor Coat Protein Target-directed Agents

Tomato chlorosis virus(ToCV),as a new virus,has caused serious losses on vegetable production in China.Due to the lack of efficient anti-ToCV commercial agent,the prevention of ToCV is so difficult to execute.How screen anti-ToCV agent is urgent scientific question.Thus,the building of anti-ToCV agent screening model and screening efficient anti-ToCV agent are significant.In this study,we considered ToCV minor coat protein(CPm)as potential drug-target to build a anti-ToCV agent screen model in vitro and in vivo.The affinity of ToCV CPm and quinazoline derivatives were tested through microscale thermophoresis(MST).And the anti-ToCV biological activity of quinazoline derivatives were evaluated by quantitative polymerase chain reaction(q-PCR).In addition,the affinity relationship of quinazoline derivatives structures and ToCV CPm was studied by three-dimensional quantitative structure-activity relationship(3D-QSAR)method.The interaction of ToCV CPm and coat protein(CP)was measured by MST and yeast two hybrid(Y2H).As shown by the results,compounds a and b exhibited the strongest affinity to ToCV CPm in vitro,with Kd value were 0.33?M and 0.55?M,respectively,which were significantly superior to Dufulin(32.31?M)and comparable to Ningnanmycin(0.45?M)and Ribavirin(0.90?M).And the compounds a and b can inhibit the ToCV CPm gene expression in vivo,their inhibit rates were 73.1%and 77.5%,which were comparable to Ningnanmycin(79.5%)and Ribavirin(78.2%).Performing Comparative Molecular Field Analysis(Co MFA)and Comparative Molecular Similarity Index Analysis(Co MSIA)methods for 3D-QSAR analysis,and effective prediction models(q~2>0.5)were constructed,which used to further revealed the relationship of the quinazoline derivatives structures and ToCV CPm affinity.Finally,the results of MST and Y2H indicated that ToCV CPm interacts with ToCV CP in vitro and in vivo.