Exceptional potency and structural basis of a T1249-derived lipopeptide fusion inhibitor against HIV-1, HIV-2, and simian immunodeficiency virus

Zhu, Yuanmei; Zhang, Xiujuan; Ding, Xiaohui; Chong, Huihui; Cui, Sheng; He, Jin‐Sheng; Wang, Xinquan; He, Yuxian

doi:10.1074/jbc.ra118.001729

Cited by 27 publications

(25 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The inhibitory activities of inhibitors on a panel of HIV-1 and SIV pseudoviruses were measured by single-cycle infection assays described previously (20,23). Briefly, a pseudovirus was generated via the cotransfection of HEK293T cells with an Env-expressing plasmid and a backbone plasmid encoding an Env-defective, luciferase-expressing HIV-1 genome (pSG3Δenv).…”

Section: Methodsmentioning

confidence: 99%

“…In short-term monotherapy, LP-19 could reduce viral loads to undetectable levels in both acutely and chronically simian-human immunodeficiency virus (SHIV) infected rhesus monkeys (20). Very recently, we developed LP-40 and LP-46 by replacing the C-terminal membrane-binding sequences of T-20 and T1249 with a fatty acid group, which sharply improved anti-HIV activity (23,24). The crystal structures of LP-40 and LP-46 in complexes with target mimic peptides revealed the key binding motifs underlying the mechanisms of action of T-20, T1249, and their lipid derivatives (23,24).…”

mentioning

confidence: 99%

“…Very recently, we developed LP-40 and LP-46 by replacing the C-terminal membrane-binding sequences of T-20 and T1249 with a fatty acid group, which sharply improved anti-HIV activity (23,24). The crystal structures of LP-40 and LP-46 in complexes with target mimic peptides revealed the key binding motifs underlying the mechanisms of action of T-20, T1249, and their lipid derivatives (23,24). In this study, we rationally designed two new lipopeptides, LP-50 and LP-51, by refining the structure and function of LP-40.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Design of Novel HIV-1/2 Fusion Inhibitors with High Therapeutic Efficacy in Rhesus Monkey Models

Chong

Xue

Zhu

et al. 2018

J Virol

Self Cite

View full text Add to dashboard Cite

T-20 (enfuvirtide) is the only approved viral fusion inhibitor that is used for the treatment of human immunodeficiency virus type 1 (HIV-1) infection; however, it has relatively low antiviral activity and easily induces drug resistance. We recently reported a T-20-based lipopeptide fusion inhibitor (LP-40) showing improved anti-HIV activity (X. Ding et al., J Virol 91:e00831-17, 2017, https://doi.org/10.1128/JVI.00831-17). In this study, we designed LP-50 and LP-51 by refining the structure and function of LP-40. The two new lipopeptides showed dramatically enhanced secondary structure and binding stability and were exceptionally potent inhibitors of HIV-1, HIV-2, simian immunodeficiency virus (SIV), and chimeric simian-human immunodeficiency virus (SHIV), with mean 50% inhibitory concentrations (ICs) in the very low picomolar range. They also exhibited dramatically increased potencies in inhibiting a panel of T-20- and LP-40-resistant mutant viruses. In line with their data, LP-50 and LP-51 exhibited extremely potent and long-lasting anti-HIV activities in rhesus monkeys: serum dilution peaks that inhibited 50% of virus infection were >15,200-fold higher than those for T-20 and LP-40. Low-dose, short-term monotherapy of LP-51 could sharply reduce viral loads to undetectable levels in acutely and chronically SHIV infected monkey models. To our knowledge, LP-50 and LP-51 are the most potent and broad HIV-1/2 and SIV fusion inhibitors, which can be developed for clinical use and can serve as tools for exploration of the mechanisms of viral entry and inhibition. T-20 remains the only membrane fusion inhibitor available for the treatment of viral infection, but its relatively low anti-HIV activity and genetic barrier for drug resistance have significantly limited its clinical application. Here we report two new lipopeptide-based fusion inhibitors (LP-50 and LP-51) showing extremely potent inhibitory activities against diverse HIV-1, HIV-2, SIV, and T-20-resistant variants. Promisingly, both inhibitors exhibited potent and long-lasting anti-HIV activity and could efficiently suppress viral loads to undetectable levels in SHIV-infected monkey models. We believe that LP-50 and LP-51 are the most potent and broad-spectrum fusion inhibitors known to date and thus have high potential for clinical development.

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Design of Novel HIV-1/2 Fusion Inhibitors with High Therapeutic Efficacy in Rhesus Monkey Models

Chong

Xue

Zhu

et al. 2018

J Virol

Self Cite

View full text Add to dashboard Cite

show abstract

“…By modifying the short peptides HP23 and 2P23 with different lipids (fatty acid, cholesterol, sphingolipids), we previously developed LP-11 and LP-19 with dramatically increased in vitro and in vivo antiviral activities and stability (35,36). To explore the mechanisms of action of lipopeptides and to develop new HIV-1 fusion inhibitors with different modes of action, we recently generated T-20-and T1249-based lipopeptides (LP-40 and LP-46) by replacing their TRM sequences with a fatty acid group, resulting in increased anti-HIV activities (37,38). Very recently, we modified LP-40 by introducing the intrahelical salt bridge-prone and HIV-2/simian immunodeficiency virus (SIV) sequences, resulting in LP-50 and LP-51, which could inhibit HIV-1, HIV-2, and SIV isolates at very low picomolar concentrations and suppress viral loads to undetectable levels in acutely and chronically simian-human immunodeficiency virus (SHIV)-infected rhesus monkey models (39).…”

mentioning

confidence: 99%

Structural and Functional Characterization of Membrane Fusion Inhibitors with Extremely Potent Activity against Human Immunodeficiency Virus Type 1 (HIV-1), HIV-2, and Simian Immunodeficiency Virus

Chong

Zhu

et al. 2018

J Virol

Self Cite

View full text Add to dashboard Cite

T-20 (enfuvirtide) is the only membrane fusion inhibitor available for the treatment of viral infection; however, it has low anti-human immunodeficiency virus (anti-HIV) activity and a low genetic barrier for drug resistance. We recently reported that T-20 sequence-based lipopeptides possess extremely potent and efficacies (X. Ding, Z. Zhang, H. Chong, Y. Zhu, H. Wei, X. Wu, J. He, X. Wang, Y. He, 2017, J Virol 91:e00831-17, https://doi.org/10.1128/JVI.00831-17; H. Chong, J. Xue, Y. Zhu, Z. Cong, T. Chen, Y. Guo, Q. Wei, Y. Zhou, C. Qin, Y. He, 2018, J Virol 92:e00775-18, https://doi.org/10.1128/JVI.00775-18). Here, we focused on characterizing the structure-activity relationships of the T-20 derivatives. First, a novel lipopeptide termed LP-52 was generated with improved target-binding stability and anti-HIV activity. Second, a large panel of truncated lipopeptides was characterized, revealing a 21-amino-acid sequence core structure. Third, it was surprisingly found that the addition of the gp41 pocket-binding residues in the N terminus of the new inhibitors resulted in increased binding but decreased antiviral activities. Fourth, while LP-52 showed the most potent activity in inhibiting divergent HIV-1 subtypes, its truncated versions, such as LP-55 (25-mer) and LP-65 (24-mer), still maintained their potencies at very low picomolar concentrations; however, both the N- and C-terminal motifs of LP-52 played crucial roles in the inhibition of T-20-resistant HIV-1 mutants, HIV-2, and simian immunodeficiency virus (SIV) isolates. Fifth, we verified that LP-52 can bind to target cell membranes and human serum albumin and has low cytotoxicity and a high genetic barrier to inducing drug resistance. Development of novel membrane fusion inhibitors against HIV and other enveloped viruses is highly important in terms of the peptide drug T-20, which remains the only one for clinical use, even if it is limited by large dosages and resistance. Here, we report a novel T-20 sequence-based lipopeptide showing extremely potent and broad activities against HIV-1, HIV-2, SIV, and T-20-resistant mutants, as well as an extremely high therapeutic selectivity index and genetic resistance barrier. The structure-activity relationship (SAR) of the T-20 derivatives has been comprehensively characterized, revealing a critical sequence core structure and the target sites of viral vulnerability that do not include the gp41 pocket. The results also suggest that membrane-anchored inhibitors possess unique modes of action relative to unconjugated peptides. Combined, our series studies have not only provided drug candidates for clinical development but also offered important tools to elucidate the mechanisms of viral fusion and inhibition.

show abstract

“…More recent efforts to take advantage of the demonstrated efficacy of these peptidyl fusion inhibitors have shown promise. For instance, conjugation of T20 and T1249 to a fatty acid moiety showed potent fusion inhibition . With new peptide delivery strategies and formulations being developed, the relevance of HIV peptidyl fusion inhibition to clinical practice may see a renaissance.…”

Section: Introductionmentioning

confidence: 99%

Optimization of peptidic HIV‐1 fusion inhibitor T20 by phage display

Chen

Cook

et al. 2019

Protein Science

View full text Add to dashboard Cite

The HIV fusion inhibitor T20 has been approved to treat those living with HIV/AIDS, but treatment gives rise to resistant viruses. Using combinatorial phage-displayed libraries, we applied a saturation scan approach to dissect the entire T20 sequence for binding to a prefusogenic five-helix bundle (5HB) mimetic of HIV-1 gp41. Our data set compares all possible amino acid substitutions at all positions, and affords a complete view of the complex molecular interactions governing the binding of T20 to 5HB. The scan of T20 revealed that 12 of its 36 positions were conserved for 5HB binding, which cluster into three epitopes: hydrophobic epitopes at the ends and a central dyad of hydrophilic residues. The scan also revealed that the T20 sequence was highly adaptable to mutations at most positions, demonstrating a striking structural plasticity that allows multiple amino acid substitutions at contact points to adapt to conformational changes, and also at noncontact points to fine-tune the interface. Based on the scan result and structural knowledge of the gp41 fusion intermediate, a library was designed with tailored diversity at particular positions of T20 and was used to derive a variant (T20v1) that was found to be a highly effective inhibitor of infection by multiple HIV-1 variants, including a common T20-escape mutant. These findings show that the plasticity of the T20 functional sequence space can be exploited to develop variants that overcome resistance of HIV-1 variants to T20 itself, and demonstrate the utility of saturation scanning for rapid epitope mapping and protein engineering.Abbreviations: 5HB, five-helix bundle; 6HB, six-helix bundle; aMLV, amphotropic murine leukemia virus; CHR, C-terminal heptad repeat; Fab, antigen-binding fragment; FP, fusion peptide; HR, heptad repeat; MPER, membrane proximal external region; NHR, N-terminal heptad repeat; wt, wild type.Additional Supporting Information may be found in the online version of this article.Significance statement: Prolonged therapeutic use of the HIV-1 fusion inhibitor T20 leads to the evolution of resistant viral strains. We surveyed comprehensively the functional contributions of all T20 residues for binding to a fusion intermediate mimic and engineered a variant T20v1 that neutralized many diverse HIV-1 strains, including a strain that was resistant to T20. These results show that combinatorial protein engineering can be applied to develop T20 variants with enhanced potencies and broadened specificities.

show abstract

Exceptional potency and structural basis of a T1249-derived lipopeptide fusion inhibitor against HIV-1, HIV-2, and simian immunodeficiency virus

Cited by 27 publications

References 59 publications

Design of Novel HIV-1/2 Fusion Inhibitors with High Therapeutic Efficacy in Rhesus Monkey Models

Design of Novel HIV-1/2 Fusion Inhibitors with High Therapeutic Efficacy in Rhesus Monkey Models

Structural and Functional Characterization of Membrane Fusion Inhibitors with Extremely Potent Activity against Human Immunodeficiency Virus Type 1 (HIV-1), HIV-2, and Simian Immunodeficiency Virus

Optimization of peptidic HIV‐1 fusion inhibitor T20 by phage display

Contact Info

Product

Resources

About