Rational Design of A Novel Small-Molecule HIV-1 Inactivator Targeting Both gp120 and gp41 of HIV-1

ABSTRACT HIV-1 replication requires direct interaction between HIV-1 reverse transcriptase (RT) and cellular eukaryotic translation elongation factor 1A (eEF1A). Our previous work showed that disrupting this interaction inhibited HIV-1 uncoating, reverse transcription, and replication, indicating its potential as an anti-HIV-1 target. In this study, we developed a sensitive, live-cell split-luciferase complementation assay (NanoBiT) to quantitatively measure inhibition of HIV-1 RT interaction with eEF1A. We used this to screen a small molecule library and discovered small-molecule oxazole-benzenesulfonamides (C7, C8, and C9), which dose dependently and specifically inhibited the HIV-1 RT interaction with eEF1A. These compounds directly bound to HIV-1 RT in a dose-dependent manner, as assessed by a biolayer interferometry (BLI) assay, but did not bind to eEF1A. These oxazole-benzenesulfonamides did not inhibit enzymatic activity of recombinant HIV-1 RT in a homopolymer assay but did inhibit reverse transcription and infection of both wild-type (WT) and nonnucleoside reverse transcriptase inhibitor (NNRTI)-resistant HIV-1 in a dose-dependent manner in HEK293T cells. Infection of HeLa cells was significantly inhibited by the oxazole-benzenesulfonamides, and the antiviral activity was most potent against replication stages before 8 h postinfection. In human primary activated CD4+ T cells, C7 inhibited HIV-1 infectivity and replication up to 6 days postinfection. The data suggest a novel mechanism of HIV-1 inhibition and further elucidate how the RT-eEF1A interaction is important for HIV-1 replication. These compounds provide potential to develop a new class of anti-HIV-1 drugs to treat WT and NNRTI-resistant strains in people infected with HIV. IMPORTANCE Antiretroviral drugs protect many HIV-positive people, but their success can be compromised by drug-resistant strains. To combat these strains, the development of new classes of HIV-1 inhibitors is essential and a priority in the field. In this study, we identified small molecules that bind directly to HIV-1 reverse transcriptase (RT) and inhibit its interaction with cellular eEF1A, an interaction which we have previously identified as crucial for HIV-1 replication. These compounds inhibit intracellular HIV-1 reverse transcription and replication of WT HIV-1, as well as HIV-1 mutants that are resistant to current RT inhibitors. A novel mechanism of action involving inhibition of the HIV-1 RT-eEF1A interaction is an important finding and a potential new way to combat drug-resistant HIV-1 strains in infected people.

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Rational Design and Identification of Harmine-Inspired, N-Heterocyclic DYRK1A Inhibitors Employing a Functional Genomic in vivo Drosophila Model System

10.26434/chemrxiv.14356946 ◽

2021 ◽

Author(s):

Brandon Ashfeld ◽

Francisco Huizar ◽

Harrison Hill ◽

Jeremiah Zartman ◽

Emily Bacher ◽

...

Keyword(s):

Small Molecule ◽

Rational Design ◽

Kinase Inhibitors ◽

Low Cost ◽

Neuronal Cell ◽

Model System ◽

In Vivo Model ◽

Functional Genomic

Deregulation of dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) plays a significant role in developmental brain defects, early-onset neurodegeneration, neuronal cell loss, and dementia. Herein, we report the discovery of three new classes of <i>N</i>-heterocyclic DYRK1A inhibitors based on the potent, yet toxic kinase inhibitors, harmine and harmol. An initial in vitro evaluation of the small molecule collection assembled revealed that the core heterocyclic motifs benzofuranones, oxindoles, and pyrrolones, showed statistically significant DYRK1A inhibition. Further, the utilization of a low cost, high-throughput functional genomic in vivo model system to identify small molecule inhibitors that normalize DYRK1A overexpression phenotypes is described. This in vivo assay confirmed the in vitro results, and the resulting correspondence validates generated classes as architectural motifs that serve as potential DYRK1A inhibitors. Further expansion and analysis of these core compound structures will allow discovery of safe, more effective chemical inhibitors of DYRK1A to ameliorate phenotypes caused by DYRK1A overexpression.

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Rational Design and Identification of Harmine-Inspired, N-Heterocyclic DYRK1A Inhibitors Employing a Functional Genomic in vivo Drosophila Model System

10.26434/chemrxiv.14356946.v1 ◽

2021 ◽

Author(s):

Brandon Ashfeld ◽

Francisco Huizar ◽

Harrison Hill ◽

Jeremiah Zartman ◽

Emily Bacher ◽

...

Keyword(s):

Small Molecule ◽

Rational Design ◽

Kinase Inhibitors ◽

Low Cost ◽

Neuronal Cell ◽

Model System ◽

In Vivo Model ◽

Functional Genomic

Deregulation of dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) plays a significant role in developmental brain defects, early-onset neurodegeneration, neuronal cell loss, and dementia. Herein, we report the discovery of three new classes of <i>N</i>-heterocyclic DYRK1A inhibitors based on the potent, yet toxic kinase inhibitors, harmine and harmol. An initial in vitro evaluation of the small molecule collection assembled revealed that the core heterocyclic motifs benzofuranones, oxindoles, and pyrrolones, showed statistically significant DYRK1A inhibition. Further, the utilization of a low cost, high-throughput functional genomic in vivo model system to identify small molecule inhibitors that normalize DYRK1A overexpression phenotypes is described. This in vivo assay confirmed the in vitro results, and the resulting correspondence validates generated classes as architectural motifs that serve as potential DYRK1A inhibitors. Further expansion and analysis of these core compound structures will allow discovery of safe, more effective chemical inhibitors of DYRK1A to ameliorate phenotypes caused by DYRK1A overexpression.

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Randomized Clinical Trial to Assess the Impact of the Broadly Neutralizing HIV-1 Monoclonal Antibody VRC01 on HIV-1 Persistence in Individuals on Effective ART

Open Forum Infectious Diseases ◽

10.1093/ofid/ofy242 ◽

2018 ◽

Vol 5 (10) ◽

Cited By ~ 11

Author(s):

Sharon A Riddler ◽

Lu Zheng ◽

Christine M Durand ◽

Justin Ritz ◽

Richard A Koup ◽

...

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Phase 1 ◽

Virus Production ◽

Plasma Viremia ◽

Double Blind ◽

Significant Difference ◽

Cd4 Binding Site ◽

The Impact ◽

Hiv 1

Abstract Background Broadly neutralizing monoclonal antibodies (bnMAbs) may promote clearance of HIV-1-expressing cells through antibody-dependent cell-mediated cytotoxicity. We evaluated the effect of the CD4-binding site bnMAb, VRC01, on measures of HIV-1 persistence in chronically infected individuals. Methods A5342 was a phase 1, randomized, double-blind, placebo-controlled, parallel-arm study. Participants on effective antiretroviral therapy (ART) were randomized to receive 2 infusions of VRC01 (40 mg/kg) at entry and week 3, and 2 infusions of placebo (saline) at weeks 6 and 9; or 2 infusions of placebo at entry and week 3, and 2 infusions of VRC01 at weeks 6 and 9. Results Infusion of VRC01 was safe and well tolerated. The median fold-change in the cell-associated HIV-1 RNA/DNA ratio from baseline to week 6 was 1.12 and 0.83 for the VRC01 and placebo arms, respectively, with no significant difference between arms (P = .16). There were no significant differences in the proportions with residual plasma viremia ≥1 copies/mL or in phorbol 12-myristate 13-acetate/ionomycin-induced virus production from CD4+ T cells between arms (both P > .05). Conclusions In individuals with chronic HIV-1 infection on ART, VRC01 infusions were safe and well tolerated but did not affect plasma viremia, cellular HIV-1 RNA/DNA levels, or stimulated virus production from CD4+ T cells. ClinicalTrials.gov Identifier NCT02411539

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A Peptide-Based HIV-1 Fusion Inhibitor with Two Tail-Anchors and Palmitic Acid Exhibits Substantially Improved In Vitro and Ex Vivo Anti-HIV-1 Activity and Prolonged In Vivo Half-Life

Molecules ◽

10.3390/molecules24061134 ◽

2019 ◽

Vol 24 (6) ◽

pp. 1134 ◽

Cited By ~ 7

Author(s):

Shan Su ◽

Giselle Rasquinha ◽

Lanying Du ◽

Qian Wang ◽

Wei Xu ◽

...

Keyword(s):

Palmitic Acid ◽

Half Life ◽

Ex Vivo ◽

Fusion Inhibitor ◽

C Terminus ◽

Further Development ◽

Anti Hiv ◽

Hiv 1

Enfuvirtide (T20) is the first U.S. FDA-approved HIV fusion inhibitor-based anti-HIV drug. Its clinical application is limited because of its low potency and short half-life. We previously reported that peptide HP23-E6-IDL, containing both N- and C-terminal anchor-tails, exhibited stronger potency and a better resistance profile than T20. Here we designed an analogous peptide, YIK, by introducing a mutation, T639I, and then a lipopeptide, YIK-C16, by adding palmitic acid (C16) at the C-terminus of YIK. We found that YIK-C16 was 4.4- and 3.6-fold more potent than HP23-E6-IDL and YIK against HIV-1IIIB infection and 13.3- and 10.5-fold more effective than HP23-E6-IDL and YIK against HIV-1Bal infection, respectively. Consistently, the ex vivo anti-HIV-1IIIB activity, as determined by the highest dilution-fold of the serum causing 50% inhibition of HIV-1 infection, of YIK-C16 in the sera of pretreated mice was remarkably higher than that of YIK or HP23-E6-IDL. The serum half-life (t1/2 = 5.9 h) of YIK-C16 was also significantly longer than that of YIK (t1/2 = 1.3 h) and HP23-E6-IDL (t1/2 = 1.0 h). These results suggest that the lipopeptide YIK-C16 shows promise for further development as a new anti-HIV drug with improved anti-HIV-1 activity and a prolonged half-life.

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HIV-1 Resistance Mechanism to an Electrostatically Constrained Peptide Fusion Inhibitor That Is Active against T-20-Resistant Strains

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00237-13 ◽

2013 ◽

Vol 57 (8) ◽

pp. 4035-4038 ◽

Cited By ~ 5

Author(s):

Kazuki Shimane ◽

Kumi Kawaji ◽

Fusako Miyamoto ◽

Shinya Oishi ◽

Kentaro Watanabe ◽

...

Keyword(s):

High Resistance ◽

Electrostatic Interactions ◽

Resistance Mechanism ◽

Cross Resistance ◽

Fusion Inhibitor ◽

Wild Type ◽

Fusion Inhibitors ◽

Resistant Strains ◽

Hiv 1 ◽

Peptide Fusion Inhibitor

ABSTRACTT-20EK is a novel fusion inhibitor designed to have enhanced α-helicity over T-20 (enfuvirtide) through engineered electrostatic interactions between glutamic acid (E) and lysine (K) substitutions. T-20EK efficiently suppresses wild-type and T-20-resistant variants. Here, we selected T-20EK-resistant variants. A combination of L33S and N43K substitutions in gp41 were required for high resistance to T-20EK. While these substitutions also caused resistance to T-20, they did not cause cross-resistance to other known fusion inhibitors.

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Perceived Burden of Informal Caregivers of a Chronically Ill Older Family Member

GeroPsych ◽

10.1024/1662-9647/a000042 ◽

2011 ◽

Vol 24 (3) ◽

pp. 143-154 ◽

Cited By ~ 20

Author(s):

Elmar Gräßel ◽

Raffaela Adabbo

Keyword(s):

Family Member ◽

Informal Caregiver ◽

Chronically Ill ◽

Stress Model ◽

The Past ◽

Perceived Burden ◽

Hypothetical Construct ◽

Care Situation ◽

The Impact ◽

Further Development

The burden of caregivers has been intensively researched for the past 30 years and has resulted in a multitude of individual findings. This review illustrates the significance of the hypothetical construct of perceived burden for the further development and design of the homecare situation. Following explanations regarding the term informal caregiver, we derive the construct burden from its conceptual association with the transactional stress model of Lazarus and Folkman. Once the extent and characteristics of burden have been set forth, we then present the impact of perceived burden as the care situation. The question of predictors of burden will lead into the last section from which implications can be derived for homecare and relief of caregivers.

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Computer simulation study about the dependence of amorphous silicon photonic waveguides efficiency on the material quality

The European Physical Journal Applied Physics ◽

10.1051/epjap/2020190250 ◽

2020 ◽

Vol 90 (3) ◽

pp. 30502

Author(s):

Alessandro Fantoni ◽

João Costa ◽

Paulo Lourenço ◽

Manuela Vieira

Keyword(s):

Amorphous Silicon ◽

High Throughput Screening ◽

Simulation Analysis ◽

Low Cost ◽

Deposition Process ◽

Large Area ◽

Sidewall Roughness ◽

Sensing Applications ◽

Fabrication Defects ◽

The Impact

Amorphous silicon PECVD photonic integrated devices are promising candidates for low cost sensing applications. This manuscript reports a simulation analysis about the impact on the overall efficiency caused by the lithography imperfections in the deposition process. The tolerance to the fabrication defects of a photonic sensor based on surface plasmonic resonance is analysed. The simulations are performed with FDTD and BPM algorithms. The device is a plasmonic interferometer composed by an a-Si:H waveguide covered by a thin gold layer. The sensing analysis is performed by equally splitting the input light into two arms, allowing the sensor to be calibrated by its reference arm. Two different 1 × 2 power splitter configurations are presented: a directional coupler and a multimode interference splitter. The waveguide sidewall roughness is considered as the major negative effect caused by deposition imperfections. The simulation results show that plasmonic effects can be excited in the interferometric waveguide structure, allowing a sensing device with enough sensitivity to support the functioning of a bio sensor for high throughput screening. In addition, the good tolerance to the waveguide wall roughness, points out the PECVD deposition technique as reliable method for the overall sensor system to be produced in a low-cost system. The large area deposition of photonics structures, allowed by the PECVD method, can be explored to design a multiplexed system for analysis of multiple biomarkers to further increase the tolerance to fabrication defects.

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Consideration of Geopolitical Challenges within the Analysis of Geoenvironmental Risks for Oil and Gas Development of the Russian Arctic

Issues of Risk Analysis ◽

10.32686/1812-5220-2019-16-6-50-59 ◽

2019 ◽

Vol 16 (6) ◽

pp. 50-59

Author(s):

O. P. Trubitsina ◽

V. N. Bashkin

Keyword(s):

Oil And Gas ◽

Oil And Gas Industry ◽

Optimal Operation ◽

The Arctic ◽

Stage Model ◽

Oil And Gas Development ◽

Gas Industry ◽

The Impact ◽

Further Development ◽

Geopolitical Risks

The article is devoted to the consideration of geopolitical challenges for the analysis of geoenvironmental risks (GERs) in the hydrocarbon development of the Arctic territory. Geopolitical risks (GPRs), like GERs, can be transformed into opposite external environment factors of oil and gas industry facilities in the form of additional opportunities or threats, which the authors identify in detail for each type of risk. This is necessary for further development of methodological base of expert methods for GER management in the context of the implementational proposed two-stage model of the GER analysis taking to account GPR for the improvement of effectiveness making decisions to ensure optimal operation of the facility oil and gas industry and minimize the impact on the environment in the geopolitical conditions of the Arctic.The authors declare no conflict of interest

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