Broad Antiviral Activity and Crystal Structure of HIV-1 Fusion Inhibitor Sifuvirtide

Xue, Yuan; Chong, Huihui; Zhang, Chao; Waltersperger, S.; Wang, Meitian; Cui, Sheng; He, Yuxian

doi:10.1074/jbc.m111.317883

Cited by 59 publications

(57 citation statements)

References 51 publications

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“…The 50% tissue culture infective dose (TCID 50 ) of a single thawed aliquot of each pseudovirus batch was determined in TZM-bl cells. The antiviral activities of duplex inhibitors were determined using TZM-bl cells as described previously (34). Briefly, the duplexes were prepared in 2-fold stepwise series dilutions, mixed with 100 TCID 50 viruses, and incubated 1 h at room temperature.…”

Section: Methodsmentioning

confidence: 99%

DNA Duplexes with Hydrophobic Modifications Inhibit Fusion between HIV-1 and Cell Membranes

Xu¹,

Cai²,

Chen

et al. 2013

Antimicrob Agents Chemother

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Discovery of new drugs for the treatment of AIDS typically possessing unique structures associated with novel mechanisms of action has been of great importance due to the quick drug-resistant mutations of HIV-1 strains. The work presented in this report describes a novel class of DNA duplex-based HIV-1 fusion inhibitors. Hydrophobic groups were introduced into a DNA duplex skeleton either at one end, at both ends, or in the middle. These modified DNA duplexes inhibited fusion between HIV-1 and human cell membranes at micro-or submicromolar concentrations. Respective inhibitors adopted an aptamer pattern instead of a base-pairing interaction pattern. Structure-activity relationship studies of the respective DNA duplexes showed that the rigid and negatively charged DNA skeletons, in addition to the presence of hydrophobic groups, were crucial to the anti-HIV-1 activity of these compounds. A fluorescent resonance energy transfer (FRET)-based inhibitory assay showed that these duplex inhibitors interacted with the primary pocket in the gp41 N-terminal heptad repeat (NHR) instead of interacting with the lipid bilayers.

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Section: Methodsmentioning

confidence: 99%

DNA Duplexes with Hydrophobic Modifications Inhibit Fusion between HIV-1 and Cell Membranes

Xu¹,

Cai²,

Chen

et al. 2013

Antimicrob Agents Chemother

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“…A panel of pseudoviruses with Envs derived from HIV-1 isolates predominantly circulating in China was generated, and previously described neutralization assay protocols were used (31,32).…”

mentioning

confidence: 99%

Exceptionally Potent and Broadly Cross-Reactive, Bispecific Multivalent HIV-1 Inhibitors Based on Single Human CD4 and Antibody Domains

Chen

Yang

Prabakaran

et al. 2014

J Virol

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Soluble forms of the human immunodeficiency virus type 1 (HIV-1) primary receptor CD4 (soluble CD4 [sCD4]) have been extensively characterized for a quarter of a century as promising HIV-1 inhibitors, but they have not been clinically successful. By combining a protein cavity-filling strategy and the power of library technology, we identified an engineered cavity-altered single-domain sCD4 (mD1.22) with a unique combination of excellent properties, including broad and potent neutralizing activity, high specificity, stability, solubility, and affinity for the HIV-1 envelope glycoprotein gp120, and small molecular size. To further improve its neutralizing potency and breadth, we generated bispecific multivalent fusion proteins of mD1.22 with another potent HIV-1 inhibitor, an antibody domain (m36.4) that targets the coreceptor-binding site on gp120. The fusion proteins neutralized all HIV-1 isolates tested, with potencies about 10-, 50-, and 200-fold higher than those of the broadly neutralizing antibody VRC01, the U.S. FDA-approved peptide inhibitor T20, and the clinically tested sCD4-Fc fusion protein CD4-Ig, respectively. In addition, they exhibited higher stability and specificity and a lower aggregation propensity than CD4-Ig. Therefore, mD1.22 and related fusion proteins could be useful for HIV-1 prevention and therapy, including eradication of the virus. Soluble forms of human CD4 (sCD4) comprising all four (D1 to D4) or the first two (D1D2) extracellular domains are potent inhibitors of the human immunodeficiency virus type 1 (HIV-1) in vitro (1, 2). Several promising monomeric (3-5), dimeric (6-8), and tetrameric (9-11) sCD4 derivatives have been tested in animal models and in human clinical trials, but they exhibited modest and transient antiviral activities. Previously, we demonstrated that decreasing the molecular size of D1D2 to a single domain, D1, significantly increased its antiviral activity and reduce its nonspecificity, i.e., interactions with molecules other than the HIV-1 envelope glycoprotein (Env) gp120; a D1 variant (mD1.2) was identified that is also more soluble than D1D2 (12). However, mD1.2 still binds to human B cells and CD4 ϩ T cells without HIV-1 Env expression, although it binds more weakly than D1D2 and its stability is comparable to that of D1D2, which is relatively low (12).It has been shown previously that some proteins exhibit poor hydrophobic packing, leading to low stability and solubility due to the presence of cavities within or on the surfaces of proteins that are either empty or hydrated (13,14). Identification of such cavities and filling them with bulkier hydrophobic amino acid side chains have proven effective in improving stability and other properties of proteins (15). By combining this cavity-filling strategy with the power of library technology, we identified an mD1.2 mutant, designated mD1.22, that has significantly higher soluble expression, thermal stability, and specificity than mD1.2. Bispecific multivalent fusion proteins of mD1.22 with m36.4, an engineered ...

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“…Twenty adult Chinese rhesus macaques were screened and found to be negative for SIV, herpesvirus B, and simian T-lymphotropic virus. SHIVs (SF162P3) were expanded on macaque peripheral blood mononuclear cells (PBMCs), and the TCID 50 was quantitated. Macaques were intravenously inoculated with 1,000 TCID 50 of viruses and randomly assigned to 5 treatment groups (4 monkeys/group).…”

Section: Methodsmentioning

confidence: 99%

“…A single-cycle infection assay was performed as described previously (50). Briefly, HIV-1 or SIV pseudoviruses were generated via the cotransfection of HEK293T cells with an Env-expressing plasmid and a backbone plasmid, pSG3Δenv, that encodes an Env-defective, luciferaseexpressing HIV-1 genome.…”

Section: Methodsmentioning

confidence: 99%

A Lipopeptide HIV-1/2 Fusion Inhibitor with Highly PotentIn Vitro,Ex Vivo, andIn VivoAntiviral Activity

Chong

Xue

Xiong

et al. 2017

J Virol

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Peptides derived from the C-terminal heptad repeat (CHR) region of the human immunodeficiency virus type 1 (HIV-1) fusogenic protein gp41 are potent viral entry inhibitors, and currently, enfuvirtide (T-20) is the only one approved for clinical use; however, emerging drug resistance largely limits its efficacy. In this study, we generated a novel lipopeptide inhibitor, named LP-19, by integrating multiple design strategies, including an N-terminal M-T hook structure, an HIV-2 sequence, intrahelical salt bridges, and a membrane-anchoring lipid tail. LP-19 showed stable binding affinity and highly potent, broad, and long-lasting antiviral activity. In in vitro studies, LP-19 efficiently inhibited HIV-1-, HIV-2-, and simian immunodeficiency virus (SIV)-mediated cell fusion, viral entry, and infection, and it was highly active against diverse subtypes of primary HIV-1 isolates and inhibitor-resistant mutants. Ex vivo studies demonstrated that LP-19 exhibited dramatically increased anti-HIV activity and an extended half-life in rhesus macaques. In short-term monotherapy, LP-19 reduced viral loads to undetectable levels in acutely and chronically simian-human immunodeficiency virus (SHIV)-infected monkeys. Therefore, this study offers an ideal HIV-1/2 fusion inhibitor for clinical development and emphasizes the importance of the viral fusion step as a drug target. IMPORTANCE The peptide drug T-20 is the only viral fusion inhibitor in the clinic, which is used for combination therapy of HIV-1 infection; however, it requires a high dosage and easily induces drug resistance, calling for a new drug with significantly improved pharmaceutical profiles. Here, we have developed a short-lipopeptide-based fusion inhibitor, termed LP-19, which mainly targets the conserved gp41 pocket site and shows highly potent inhibitory activity against HIV-1, HIV-2, and even SIV isolates. LP-19 exhibits dramatically increased antiviral activity and an extended half-life in rhesus macaques, and it has potent therapeutic efficacy in SHIV-infected monkeys, highlighting its high potential as a new viral fusion inhibitor for clinical use.

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Broad Antiviral Activity and Crystal Structure of HIV-1 Fusion Inhibitor Sifuvirtide

Cited by 59 publications

References 51 publications

DNA Duplexes with Hydrophobic Modifications Inhibit Fusion between HIV-1 and Cell Membranes

DNA Duplexes with Hydrophobic Modifications Inhibit Fusion between HIV-1 and Cell Membranes

Exceptionally Potent and Broadly Cross-Reactive, Bispecific Multivalent HIV-1 Inhibitors Based on Single Human CD4 and Antibody Domains

A Lipopeptide HIV-1/2 Fusion Inhibitor with Highly PotentIn Vitro,Ex Vivo, andIn VivoAntiviral Activity

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