Specific Inhibitors of HIV Capsid Assembly Binding to the C-Terminal Domain of the Capsid Protein: Evaluation of 2-Arylquinazolines as Potential Antiviral Compounds

The assembly of immature retroviral particles is initiated in the cytoplasm by the binding of the structural polyprotein precursor Gag with viral genomic RNA. The protein interactions necessary for assembly are mediated predominantly by the capsid (CA) and nucleocapsid (NC) domains, which have conserved structures. In contrast, the structural arrangement of the CA-NC connecting region differs between retroviral species. In HIV-1 and Rous sarcoma virus, this region forms a rod-like structure that separates the CA and NC domains, whereas in Mason-Pfizer monkey virus, this region is densely packed, thus holding the CA and NC domains in close proximity. Interestingly, the sequence connecting the CA and NC domains in gammaretroviruses, such as murine leukemia virus (MLV), is unique. The sequence is called a charged assembly helix (CAH) due to a high number of positively and negatively charged residues. Although both computational and deletion analyses suggested that the MLV CAH forms a helical conformation, no structural or biochemical data supporting this hypothesis have been published. Using an in vitro assembly assay, alanine scanning mutagenesis, and biophysical techniques (circular dichroism, NMR, microcalorimetry, and electrophoretic mobility shift assay), we have characterized the structure and function of the MLV CAH. We provide experimental evidence that the MLV CAH belongs to a group of charged, E(R/K)-rich, single ␣-helices. This is the first single ␣-helix motif identified in viral proteins.

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“…Additionally, we used HIV-1 CA, prepared as described in Ref. 34, as a control. We performed ITC dilution experiments (Fig.…”

Section: Resultsmentioning

confidence: 99%

Functional and Structural Characterization of Novel Type of Linker Connecting Capsid and Nucleocapsid Protein Domains in Murine Leukemia Virus

Doležal

Hadravová

Kožíšek

et al. 2016

Journal of Biological Chemistry

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“…There are few reports of non-peptidic compounds targeting CTD of CA. [7][8][9][10][11] The other reported compounds bind to the NTD of CA and some of them affect capsid assembly. Three independent binding sites have been defined in the NTD.…”

Section: Introductionmentioning

confidence: 99%

“…7 Because the inhibitors compete with CAI, we proposed that they bind into a conserved hydrophobic groove in CA CTD. 20 We further optimized the identified hits to improve the inhibitory activity, resulting in the compounds shown in Figure 1.…”

Section: Introductionmentioning

confidence: 99%

“…These compounds exhibit moderate affinity to the binding site in vitro, as determined by AlphaScreen assay and ITC. 7 To improve the pharmacokinetics and binding of those quinazoline compounds, we searched for derivatives with superior solubility. From previous work, 7 it was obvious that the 4-N-phenyl and 2-pyridinyl moieties are crucial for binding.…”

Section: Introductionmentioning

confidence: 99%

“…7 To improve the pharmacokinetics and binding of those quinazoline compounds, we searched for derivatives with superior solubility. From previous work, 7 it was obvious that the 4-N-phenyl and 2-pyridinyl moieties are crucial for binding. Therefore, we decided to replace the parent quinazoline scaffold with a smaller, and less hydrophobic, heterocyclic pyrimidine.…”

Section: Introductionmentioning

confidence: 99%

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Synthesis and evaluation of 2-pyridinylpyrimidines as inhibitors of HIV-1 structural protein assembly

Kožíšek

Štěpánek

Parkan

et al. 2016

Bioorganic & Medicinal Chemistry Letters

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In an effort to identify an HIV-1 capsid assembly inhibitor with improved solubility and potency, we synthesized two series of pyrimidine analogues based on our earlier lead compound N-(4-(ethoxycarbonyl)phenyl)-2-(pyridine-4-yl)quinazoline-4-amine. In vitro binding experiments showed that our series of 2-pyridine-4-ylpyrimidines had IC50 values higher than 28μM. Our series of 2-pyridine-3-ylpyrimidines exhibited IC50 values ranging from 3 to 60μM. The congeners with a fluoro substituent introduced at the 4-N-phenyl moiety, along with a methyl at C-6, represent potent HIV capsid assembly inhibitors binding to the C-terminal domain of the capsid protein.

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“On‐Water” Palladium‐Catalyzed Tandem Cyclization Reaction for the Synthesis of Biologically Relevant 4‐Arylquinazolines

Yuan

Liu

2019

Chemistry A European J

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The quinazoline scaffold is prevalenti np harmaceutically relevant molecules that show diverse biological activities. Herein, we report an efficient" on-water" palladium-catalyzedt andem cyclization reaction from commercially availablea rylboronic acids andb enzonitriles that enable the rapid access to 4-arylquinazoline scaffolds in good to excellent yields (45 examples, up to 98 %y ield). This protocol has shown good functional group tolerance and broad substrate scope. The reactionw as also performed on ag ram scale ands uccessfully appliedt ot he synthesis of the highlyp otent and selective PI3Kd inhibitor N11, showingt he practicability and synthetic utility of the protocol. In this reaction, the quinazoline scaffold is efficientlyc onstructed with the simultaneous formationo f one CÀCb ond ando ne CÀNb ond. Collectively,t he protocol could serve as an alternative strategy to synthesize biologically important quinazoline scaffolds.Supporting information and the ORCID identification number(s) for the author(s) of this articlecan be found under: https://doi.

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Specific Inhibitors of HIV Capsid Assembly Binding to the C-Terminal Domain of the Capsid Protein: Evaluation of 2-Arylquinazolines as Potential Antiviral Compounds

Cited by 41 publications

References 35 publications

Functional and Structural Characterization of Novel Type of Linker Connecting Capsid and Nucleocapsid Protein Domains in Murine Leukemia Virus

Functional and Structural Characterization of Novel Type of Linker Connecting Capsid and Nucleocapsid Protein Domains in Murine Leukemia Virus

Synthesis and evaluation of 2-pyridinylpyrimidines as inhibitors of HIV-1 structural protein assembly

“On‐Water” Palladium‐Catalyzed Tandem Cyclization Reaction for the Synthesis of Biologically Relevant 4‐Arylquinazolines

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