• Alzheimer’s disease
• ISR
• dark microglia
• integrated stress response
• lipid secretion
• lipotoxicity
• microglia
• neurodegeneration
• neurotoxic microglia
• non-cell-autonomous stress

• Microglia/metabolism
• Microglia/pathology
• Microglia/ultrastructure
• Animals
• Mice
• Alzheimer Disease/pathology
• Alzheimer Disease/metabolism
• Mice, Transgenic
• Lipids/toxicity
• Stress, Physiological/physiology
• Neurons/metabolism
• Neurons/pathology
• Mice, Inbred C57BL
• Humans
• Synapses/pathology
• Synapses/metabolism
• Lipid Metabolism

• drugs
• precursors
• pyrrolidine

• Pyrrolidines/chemistry
• Pyrrolidines/chemical synthesis
• Stereoisomerism
• Cyclization
• Chemistry Techniques, Synthetic/methods

• AP23488720 Science Committee of the Ministry of Science and Higher Education of the Republic of Kazakhstan

• CAR-T cell therapy
• solid cancers
• synthetic biology

• Humans
• Neoplasms/therapy
• Neoplasms/immunology
• Synthetic Biology/methods
• Immunotherapy, Adoptive/methods
• Receptors, Chimeric Antigen/immunology
• Receptors, Chimeric Antigen/genetics
• T-Lymphocytes/immunology
• Antigens, Neoplasm/immunology

• Clinical applications
• Drugs
• Hepatitis
• Small-molecule
• Synthesis

• Humans
• Carcinoma, Hepatocellular/drug therapy
• Liver Neoplasms/drug therapy
• Antiviral Agents/pharmacology
• Antiviral Agents/therapeutic use
• Hepatitis B/drug therapy
• Hepatitis B virus

• Structure Activity Relationship (SAR)
• antiviral agents
• heterocycles
• natural products
• synthetic methods

• Antiviral Agents/pharmacology
• Drug Discovery
• Structure-Activity Relationship

• Antiviral Agents
• Malignancy
• Oncoviruses
• Therapeutics
• Viral Oncogenesis

• Humans
• Epstein-Barr Virus Infections/drug therapy
• Herpesvirus 4, Human
• Carcinogenesis
• Neoplasms/drug therapy
• Neoplasms/prevention & control
• Neoplasms/pathology
• Antiviral Agents/pharmacology
• Antiviral Agents/therapeutic use

• Binding free energy calculation
• Hepatitis C virus
• Molecular docking
• Molecular dynamics simulations
• NS3/4A protease
• Small molecule inhibitors

• Humans
• Hepacivirus
• Molecular Dynamics Simulation
• Molecular Docking Simulation
• Protease Inhibitors/pharmacology
• Viral Nonstructural Proteins/chemistry
• Hepatitis C/drug therapy
• Antiviral Agents/pharmacology
• Antiviral Agents/chemistry

• P41 GM103311 NIGMS NIH HHS
• R41 AI112114 NIAID NIH HHS

• Isoquinoline
• activity
• chemical property
• clinical drug
• disease
• pharmacology

• bile acids
• bile salt export pump (BSEP)
• biomarkers
• knockdown
• liquid chromatography-mass spectrometry (LC-MS)
• microbiome
• transporters

New protein synthesis is known to be required for the consolidation of memories, yet existing methods to block translation lack spatiotemporal precision and cell-type specificity, preventing investigation of cell-specific contributions of protein synthesis. Here, we developed a combined knock-in mouse and chemogenetic approach for cell type-specific and drug-inducible protein synthesis inhibition (ciPSI) that enables rapid and reversible phosphorylation of eIF2α, leading to inhibition of general translation by 50% in vivo. We use ciPSI to show that targeted protein synthesis inhibition pan-neuronally and in excitatory neurons in lateral amygdala (LA) impaired long-term memory. This could be recovered with artificial chemogenetic activation of LA neurons, though at the cost of stimulus generalization. Conversely, genetically reducing phosphorylation of eIF2α in excitatory neurons in LA enhanced memory strength, but reduced memory fidelity and behavioral flexibility. Our findings provide evidence for a cell-specific translation program during consolidation of threat memories.

The discovery of asunaprevir (1) began with the concept of engaging the small and well-defined S₁’ pocket of the hepatitis C virus (HCV) NS3/4A protease that was explored in the context of tripeptide carboxylic acid-based inhibitors. A cyclopropyl-acyl sulfonamide moiety was found to be the optimal element at the P₁-P₁’ interface enhancing the potency of carboxylic acid-based prototypes by 10- to >100-fold, dependent upon the specific background. Optimization for oral bioavailability identified a 1-substituted isoquinoline-based P₂* element that conferred a significant exposure advantage in rats compared to the matched 4-substituted quinoline isomer. BMS-605339 (30) was the first cyclopropyl-acyl sulfonamide derivative advanced into clinical trials that demonstrated dose-related reductions in plasma viral RNA in HCV-infected patients. However, 30 was associated with cardiac events observed in a normal healthy volunteer (NHV) and an HCV-infected patient that led to the suspension of the development program. Using a Langendorff rabbit heart model, a limited structure-cardiac liability relationship was quickly established that led to the discovery of 1. This compound, which differs from 30 only by changes in the substitution pattern of the P₂* isoquinoline heterocycle and the addition of a single chlorine atom to the molecular formula, gave a dose-dependent reduction in plasma viral RNA following oral administration to HCV-infected patients without the burden of the cardiac events that had been observed with 30. A small clinical trial of the combination of 1 with the HCV NS5A inhibitor daclatasvir (2) established for the first time that a chronic genotype 1 (GT-1) HCV infection could be cured by therapy with two direct-acting antiviral agents in the absence of exogenous immune-stimulating agents. Development of the combination of 1 and 2 was initially focused on Japan where the patient population is predominantly infected with GT-1b virus, culminating in marketing approval which was granted on July 4, 2014. In order to broaden therapy to include GT-1a infections, a fixed dose triple combination of 1, 2, and the allosteric NS5B inhibitor beclabuvir (3) was developed, approved by the Japanese health authorities for the treatment of HCV GT-1 infection on December 20, 2016 and marketed as Ximency^®.

• Direct-acting antivirals
• HCV
• glecaprevir
• pangenotypic drugs
• pibrentasvir
• real-world
• sofosbuvir
• velpatasvir
• voxilaprevir.

• Antiviral Agents/therapeutic use
• Genotype
• Hepatitis C, Chronic/drug therapy
• Humans
• Sustained Virologic Response

To investigate the specific effects of immunosuppressants on the antiviral action of daclatasvir and asunaprevir.

The antiviral activity of daclatasvir (DCV) and asunaprevir (ASV) combined with immunosuppressants was tested using two in vitro models for hepatitis C virus (HCV) infection.

Tacrolimus, rapamycin and cyclosporine did not negatively affect the antiviral action of DCV or ASV. Mycophenolic acid (MPA) showed additive antiviral effects combined with these direct acting antivirals (DAAs). MPA induces interferon-stimulated genes (ISGs) and is a potent GTP synthesis inhibitor. DCV or ASV did not induce ISGs expression nor affected ISG induction by MPA. Rather, the combined antiviral effect of MPA with DCV and ASV was partly mediated via inhibition of GTP synthesis.

Immunosuppressants do not negatively affect the antiviral activity of DAAs. MPA has additive effect on the antiviral action of DCV and ASV. This combined benefit needs to be confirmed in prospective clinical trials.

• Immunosuppressant
• Hepatitis C
• Daclatasvir
• Asunaprevir
• Liver
• Transplantation

Asunaprevir (ASV) is a potent, pangenotypic, twice-daily hepatitis C virus (HCV) NS3 inhibitor indicated for the treatment of chronic HCV infection.

A population pharmacokinetic (PPK) model was developed using pooled ASV concentration data from 1239 HCV-infected subjects who received ASV either as part of the DUAL regimen with daclatasvir or as part of the QUAD regimen with daclatasvir and peg-interferon/ribavirin.

A two-compartment model with first-order elimination from the central compartment, an induction effect on clearance, and an absorption model consisted of zero-order release followed by first-order absorption adequately described ASV PK after oral administration. A typical value for ASV clearance (CL/F) was 50.8 L/h, increasing by 43% after 2 days to a CL/F of 72.5 L/h at steady-state, likely due to auto-induction of cytochrome P450 3A4 (CYP3A4). Factors indicative of hepatic function were identified as key influential covariates on ASV exposures. Subjects with cirrhosis had an 84% increase in ASV area under the concentration time curve (AUC) and subjects with baseline aspartate aminotransferase (AST) above 78 IU/L had a 58% increase in area under the concentration time curve (AUC). Asians subjects had a 46% higher steady-state AUC relative to White/Caucasian subjects. Other significant covariates were formulation, age, and gender.

The current PPK model provided a parsimonious description of ASV concentration data in HCV-infected subjects. Key covariates identified in the model help explain the observed variability in ASV exposures and may guide clinical use of the drug.

Bristol-Myers Squibb.

The online version of this article (10.1007/s40121-018-0197-y) contains supplementary material, which is available to authorized users.

• Asunaprevir
• Direct-acting antivirals
• Hepatitis C infection
• NS3 protease inhibitors
• Population pharmacokinetics

• Bristol-Myers Squibb Foundation

Viruses of the flaviviridae family are responsible for some of the major infectious viral diseases around the world and there is an urgent need for drug development for these diseases. Most of the virtual screening methods in flaviviral drug discovery suffer from a low hit rate, strain-specific efficacy differences, and susceptibility to resistance. It is because they often fail to capture the key pharmacological features of the target active site critical for protein function inhibition. So in our current work, for the flaviviral NS3 protease, we summarized the pharmacophore features at the protease active site as anchors (subsite-moiety interactions).

For each of the four flaviviral NS3 proteases (i.e., HCV, DENV, WNV, and JEV), the anchors were obtained and summarized into ‘Pharmacophore anchor (PA) models’. To capture the conserved pharmacophore anchors across these proteases, were merged the four PA models. We identified five consensus core anchors (CEH1, CH3, CH7, CV1, CV3) in all PA models, represented as the “Core pharmacophore anchor (CPA) model” and also identified specific anchors unique to the PA models. Our PA/CPA models complied with 89 known NS3 protease inhibitors. Furthermore, we proposed an integrated anchor-based screening method using the anchors from our models for discovering inhibitors. This method was applied on the DENV NS3 protease to screen FDA drugs discovering boceprevir, telaprevir and asunaprevir as promising anti-DENV candidates. Experimental testing against DV2-NGC virus by in-vitro plaque assays showed that asunaprevir and telaprevir inhibited viral replication with EC₅₀ values of 10.4 μM & 24.5 μM respectively. The structure-anchor-activity relationships (SAAR) showed that our PA/CPA model anchors explained the observed in-vitro activities of the candidates. Also, we observed that the CEH1 anchor engagement was critical for the activities of telaprevir and asunaprevir while the extent of inhibitor anchor occupation guided their efficacies.

These results validate our NS3 protease PA/CPA models, anchors and the integrated anchor-based screening method to be useful in inhibitor discovery and lead optimization, thus accelerating flaviviral drug discovery.

The online version of this article (10.1186/s12859-017-1957-5) contains supplementary material, which is available to authorized users.

• Core and specific anchors
• DENV NS3 protease
• Flaviviral NS3 proteases
• Integrated anchor-based virtual screening
• Pharmacophore anchor models

• HCC
• clinical drug trial
• hepatocellular carcinoma
• management
• molecular targeted therapies
• novel therapeutics
• treatment

• Antiviral Agents/therapeutic use
• Hepatitis C, Chronic/drug therapy
• Humans
• Liver Cirrhosis/drug therapy
• Protease Inhibitors/therapeutic use

• BMS-650032
• DAA
• HCV
• NS3 protease
• beclabuvir
• daclatasvir
• interferon-free

• BMS-791325
• IFN-free combinations
• NS5B polymerase inhibitors
• allosteric
• antivirals
• direct-acting antiviral agent
• eradication
• hepatitis C virus

• antiviral resistance
• asunaprevir
• combination therapy
• daclatasvir
• deep sequencing

More than twenty years of study has provided a better understanding of hepatitis C virus (HCV) life cycle, including the general properties of viral RNA and proteins. This effort facilitates the development of sensitive diagnostic tools and effective antiviral treatments. At present, serologic screening test is recommended to perform on individuals in the high risk groups and nucleic acid tests are recommended to confirm the active HCV infections. Quantization and genotyping of HCV RNAs are important to determine the optimal duration of anti-viral therapy and predict the likelihood of response. In the early 2000s, pegylated interferon plus ribavirin became the standard anti-HCV treatment. However, this therapy is not ideal. To 2014, boceprevir, telaprevir, simeprevir, sofosbuvir and Harvoni are approved by Food and Drug Administration for the treat of HCV infections. It is likely that the new all-oral, interferon-free, pan-genotyping anti-HCV therapy will be available within the next few years. Majority of HCV infections will be cured by these anti-viral treatments. However, not all patients are expected to be cured due to viral resistance and the high cost of antiviral treatments. Thus, an efficient prophylactic vaccine will be the next challenge in the fight against HCV infection.

• Hepatitis C virus
• Diagnosis
• Treatment
• Hepatocellular carcinoma
• Nucleic acid test
• Enzyme immunoassay
• Interferon
• Direct acting antivirals
• Host-targeted agents
• Sofosbuvir

• NS3 protease inhibitor
• asunaprevir
• direct-acting antivirals
• hepatitis C virus
• interferon
• ribavirin

• GS-9669
• interferon-free
• ledipasvir
• pegylated-interferon
• resistance
• ribavirin
• sofosbuvir