A HTRF based competitive binding assay for screening specific inhibitors of HIV-1 capsid assembly targeting the C-Terminal domain of capsid

Zhang, Dawei; Luo, Rong-Hua; Xu, Lei; Yang, Liu‐Meng; Xu, Xiaojun; Bedwell, Gregory J.; Engelman, Alan; Zheng, Yong‐Tang; Chang, Shan

doi:10.1016/j.antiviral.2019.104544

Cited by 9 publications

(8 citation statements)

References 44 publications

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“…We have previously developed and validated an HTRF-based assay for the screening of inhibitors that target the CAI-binding pocket. Using this method, three compounds were identified as binding to this cavity and demonstrated their inhibitory effects against HIV-1 replication, thereby validating the efficiency of the developed HTRF assay in identifying the assembly inhibitors ( 37 , 38 ). In the present study, we used the developed assay to screen a library in order to identify compounds that may interact with the CAI-binding pocket in CTD.…”

Section: Discussionmentioning

confidence: 75%

“…Recombinant wild-type (WT) and mutant HIV-1 HXB2 Cas were produced using a previously described method ( 57 ). The C-terminal domain of CA, expressed as an N-terminal glutathione S -transferase (GST) fusion protein, was purified as previously described ( 25 , 37 ). CA hexamers were obtained by introducing quadruple mutations (A14C/E45C/W184A/M185A) as previously described ( 58 ).…”

Section: Methodsmentioning

confidence: 99%

“…The protein and the peptide were also diluted in the reaction buffer. To perform the primary screening, a previously developed method was used ( 37 ). Briefly, one micromole of each compound (i.e., 2 μL CAI-biotin peptide and 2 μL GST-CA CTD) were dispensed into 384-well white shallow microplates.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

A Broad-Spectrum Antiviral Molecule, Protoporphyrin IX, Acts as a Moderator of HIV-1 Capsid Assembly by Targeting the Capsid Hexamer

Zhang

Xie

et al. 2023

Microbiol Spectr

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Section: Discussionmentioning

confidence: 75%

Section: Methodsmentioning

confidence: 99%

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A Broad-Spectrum Antiviral Molecule, Protoporphyrin IX, Acts as a Moderator of HIV-1 Capsid Assembly by Targeting the Capsid Hexamer

Zhang

Xie

et al. 2023

Microbiol Spectr

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“…Screening of the library with 464 protein kinase inhibitors led to the identification of TX-1918 (Table 1), which inhibited the processing of Gag polyprotein in a dose-dependent manner. It also inhibited the HIV-1 CA assembly in vitro as observed by changes in absorbance [79].…”

Section: Other Inhibitorsmentioning

confidence: 90%

Targeting the Virus Capsid as a Tool to Fight RNA Viruses

Hozáková

Vokatá

Ruml

et al. 2022

Viruses

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Several strategies have been developed to fight viral infections, not only in humans but also in animals and plants. Some of them are based on the development of efficient vaccines, to target the virus by developed antibodies, others focus on finding antiviral compounds with activities that inhibit selected virus replication steps. Currently, there is an increasing number of antiviral drugs on the market; however, some have unpleasant side effects, are toxic to cells, or the viruses quickly develop resistance to them. As the current situation shows, the combination of multiple antiviral strategies or the combination of the use of various compounds within one strategy is very important. The most desirable are combinations of drugs that inhibit different steps in the virus life cycle. This is an important issue especially for RNA viruses, which replicate their genomes using error-prone RNA polymerases and rapidly develop mutants resistant to applied antiviral compounds. Here, we focus on compounds targeting viral structural capsid proteins, thereby inhibiting virus assembly or disassembly, virus binding to cellular receptors, or acting by inhibiting other virus replication mechanisms. This review is an update of existing papers on a similar topic, by focusing on the most recent advances in the rapidly evolving research of compounds targeting capsid proteins of RNA viruses.

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“…High potency, low toxicity and effectiveness against drug-resistant variants are desirable characteristics for newly developed antiretroviral drugs, that should be also amenable to long-acting therapy and increased dosing intervals. Islatravir (a novel NRTI) and lenacapavir (an HIV capsid assembly inhibitor) are the most advanced candidates, and their antiretroviral potential is being evaluated in phase 3 clinical trials, both in classical and long-acting formulations [48,49].…”

Section: Novel Antiretroviral Drugs In Advanced Clinical Trialsmentioning

confidence: 99%