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At least 271 records · Page 15

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NASA NTRS1999

Dynamic Deformation of ETNA Volcano Observed by GPS and SAR Interferometry

Synthetic aperture radar (SAR) interferometry and GPS have shown that during the quiescent period from 1993-1995 Mt. Etna volcano, Italy, inflated. Since the initiation of eruptive activity since late 1995 the deformation has been more contentious. We will explore the detailed deformation during the period from 1995-1996 spanning the late stages of inflation and the beginning of eruptive activity. We use SAR interferometry and GPS data to measure the volcano deformation. We invert the observed deformation for both simple point source. le crack elastic sources or if warranted for a spheroidal pressure So In particular, we will examine the evolution of the inflation and the transition to a lesser deflation observed at the end of 1995. We use ERS-1/2 SAR data from both ascending and descending passes to allow for dense temporal 'sampling of the deformation and to allow us to critically assess atmospheric noise. Preliminary results from interferometry suggest that the inflation rate accelerated prior to resumption of activity in 1995, while GPS data suggest a more steady inflation with some fluctuation following the start of activity. This study will compare and contrast the interferometric SAR and GPS results and will address the strengths and weaknesses of each technique towards volcano deformation studies.

Lundgren, P.↗

NASA NTRS2000

Internal Wave Study in the South China Sea Using SAR

Recently, the internal wave distribution maps in the China Seas have been compiled from hundreds of ERS-1/2, RADARSAT, and Space Shuttle SAR (Synthetic Aperture Radar) images from 1993 to 1999. Based on internal wave distribution map, most of internal waves in the northeast part of South China Sea were propagating westward. The wave crest can be as long as 200 km with amplitude of 100 m due to strong current from the Kuroshio branching out into the South China Sea. Based on the observations from drilling rigs near DongSha Island by Amoco Production Co., the solitons may be generated in a 4 km wide channel between Batan and Sabtang islands in Luzon Strait. The proposed generation mechanism is similar to the lee wave formation from a shallow topography. Both depression and elevation internal waves have been observed in the same RADARSAT ScanSAR image on May 4, 1998 near DongSha Island. Furthermore, depression and elevation internal waves have also been observed by SAR at the same location on the shelf in April and June, 1993 (in different seasons) respectively. Numerical models have been used to interpret their generation mechanism and evolution processes. Based on the SAR images, near DongSha Island, the westward propagating huge internal solitons are often encountered and diffracted/broken by the coral reefs on the shelf. After passing the island, the diffracted waves will re-merge or interact with each other. It has been observed that after the nonlinear wave-wave interaction, the phase of wave packet is shifted and wavelength is also changed. Examples of mesoscale features observed in SAR images, such as fronts, raincells, bathymetry, ship wakes, and oil spills will be presented. Recent mooring measurements in April 1999 near Dongsha Island, future field test ASIAEX (Asian Seas International Acoustics Experiment) planned for April 2001, and some pretest survey data will be discussed in this paper.

Liu, Antony K.↗

NASA NTRS2001

Validating the use of Satellite SAR for Measuring Small-Scale Surface Wind Variations

We present aircraft measurements of near-surface atmospheric boundary layer roll signatures and radar-derived sea surface roughness. The open-ocean data were collected at an altitude of 20 m using NOAA's Long-EZ aircraft. In addition, the flight was coincident with a RADARSAT SAR overpass where presumed boundary layer roll impacts are prevalent in the SAR backscatter image. The Long-EZ air and sea data are completely coincident in space and time and this unique feature allows us to validate the supposed link between SAR backscatter signatures and organized large eddy surface impacts. We find remarkable correlation between measured modulations in the along-wind component of wind speed and radar backscatter for the atmospheric eddy scale of 1 to 1.5 km. Close agreement between normalized modulation amplitudes suggests the radar-inferred surface slope variance is changing linearly with wind speed. Implications for SAR usage in coastal regions and for boundary layer study will be discussed.

Vandemark, Douglas↗

NASA NTRS1993

The Effects of Noise on Polarimetric SAR Data

Polarimetric SAR data can provide a great deal of information about the scattering behavior of the surface under observation. Polarimetric SAR systems often measure the scattering matrices of the areas under observation in linear polarizations (H and V). From the scattering matrix commonly used forms such as the covariance matrix and the Stokes matrix can be easily derived. Other measures derived from polarimetric SAR data include correlation coefficients between scattering matrix terms and the mode and variance of phase differences between scattering matrix terms. The effects of additive system noise on these measurements is not often considered in the literature on this subject. In this paper, the effects of additive system noise on measurements derived from polarimetric SAR data will be examined. It will be shown how first-order noise effects can be removed and how second-order noise effects can be reduced for some measurements...

Freeman, A.↗

NASA NTRS2006

High-Level Performance Modeling of SAR Systems

SAUSAGE (Still Another Utility for SAR Analysis that s General and Extensible) is a computer program for modeling (see figure) the performance of synthetic- aperture radar (SAR) or interferometric synthetic-aperture radar (InSAR or IFSAR) systems. The user is assumed to be familiar with the basic principles of SAR imaging and interferometry. Given design parameters (e.g., altitude, power, and bandwidth) that characterize a radar system, the software predicts various performance metrics (e.g., signal-to-noise ratio and resolution). SAUSAGE is intended to be a general software tool for quick, high-level evaluation of radar designs; it is not meant to capture all the subtleties, nuances, and particulars of specific systems. SAUSAGE was written to facilitate the exploration of engineering tradeoffs within the multidimensional space of design parameters. Typically, this space is examined through an iterative process of adjusting the values of the design parameters and examining the effects of the adjustments on the overall performance of the system at each iteration. The software is designed to be modular and extensible to enable consideration of a variety of operating modes and antenna beam patterns, including, for example, strip-map and spotlight SAR acquisitions, polarimetry, burst modes, and squinted geometries.

Chen, Curtis↗

NASA NTRS2004

Software for Generating Strip Maps from SAR Data

Jurassicprok is a computer program that generates strip-map digital elevation models and other data products from raw data acquired by an airborne synthetic-aperture radar (SAR) system. This software can process data from a variety of airborne SAR systems but is designed especially for the GeoSAR system, which is a dual-frequency (P- and X-band), single-pass interferometric SAR system for measuring elevation both at the bare ground surface and top of the vegetation canopy. Jurassicprok is a modified version of software developed previously for airborne-interferometric- SAR applications. The modifications were made to accommodate P-band interferometric processing, remove approximations that are not generally valid, and reduce processor-induced mapping errors to the centimeter level. Major additions and other improvements over the prior software include the following: a) A new, highly efficient multi-stage-modified wave-domain processing algorithm for accurately motion compensating ultra-wideband data; b) Adaptive regridding algorithms based on estimated noise and actual measured topography to reduce noise while maintaining spatial resolution; c) Exact expressions for height determination from interferogram data; d) Fully calibrated volumetric correlation data based on rigorous removal of geometric and signal-to-noise decorrelation terms; e) Strip range-Doppler image output in user-specified Doppler coordinates; f) An improved phase-unwrapping and absolute-phase-determination algorithm; g) A more flexible user interface with many additional processing options; h) Increased interferogram filtering options; and i) Ability to use disk space instead of random- access memory for some processing steps.

Hensley, Scott↗

NASA NTRS2011

Wavelet Filter Banks for Super-Resolution SAR Imaging

This paper discusses Innovative wavelet-based filter banks designed to enhance the analysis of super resolution Synthetic Aperture Radar (SAR) images using parametric spectral methods and signal classification algorithms, SAR finds applications In many of NASA's earth science fields such as deformation, ecosystem structure, and dynamics of Ice, snow and cold land processes, and surface water and ocean topography. Traditionally, standard methods such as Fast-Fourier Transform (FFT) and Inverse Fast-Fourier Transform (IFFT) have been used to extract Images from SAR radar data, Due to non-parametric features of these methods and their resolution limitations and observation time dependence, use of spectral estimation and signal pre- and post-processing techniques based on wavelets to process SAR radar data has been proposed. Multi-resolution wavelet transforms and advanced spectral estimation techniques have proven to offer efficient solutions to this problem.

Sheybani, Ehsan O.↗

NASA NTRS2019

The SAR Handbook: Comprehensive Methodologies for Forest Monitoring and Biomass Estimation

This Synthetic Aperture Radar (SAR) handbook of applied methods for forest monitoring and biomass estimation has been developed by SERVIR in collaboration with SilvaCarbon to address pressing needs in the development of operational forest monitoring services. Despite the existence of SAR technology with all-weather capability for over 30 years, the applied use of this technology for operational purposes has proven difficult. This handbook seeks to provide understandable, easy-to-assimilate technical material to remote sensing specialists that may not have expertise on SAR but are interested in leveraging SAR technology in the forestry sector.

Flores-Anderson, Africa Ixmuca↗

NASA NTRS2021

The NASA AfriSAR Campaign: Airborne SAR and Lidar Measurements of Tropical Forest Structure and Biomass in Support of Current and Future Space Missions

In 2015 and 2016, the AfriSAR campaign was carried out as a collaborative effort among international space and National Park agencies (ESA, NASA, ONERA, DLR, ANPN and AGEOS) in support of the upcoming ESA BIOMASS, NASA-ISRO Synthetic Aperture Radar (NISAR) and NASA Global Ecosystem Dynamics Initiative (GEDI) missions. The NASA contribution to the campaign was conducted in 2016 with the NASA LVIS (Land Vegetation and Ice Sensor) Lidar, the NASA L-band UAVSAR (Uninhabited Aerial Vehicle Synthetic Aperture Radar). A central motivation for the AfriSAR deployment was the common AGBD estimation requirement for the three future spaceborne missions, the lack of sufficient airborne and ground calibration data covering the full range of ABGD in tropical forest systems, and the intercomparison and fusion of the technologies. During the campaign, over 7000 km2 of waveform Lidar data from LVIS and 30000 km2 of UAVSAR data were collected over 10 key sites and transects. In addition, field measurements of forest structure and biomass were collected in sixteen 1 hectare sized plots. The campaign produced gridded Lidar canopy structure products, gridded aboveground biomass and associated uncertainties, Lidar based vegetation canopy cover profile products, Polarimetric Interferometric SAR and Tomographic SAR products and field measurements. Our results showcase the types of data products and scientific results expected from the spaceborne Lidar and SAR missions; we also expect that the AfriSAR campaign data will facilitate further analysis and use of waveform Lidar and multiple baseline polarimetric SAR datasets for carbon cycle, biodiversity, water resources and more applications by the greater scientific community.

AfriSAR↗

NASA NTRS2021

Comparison of SAR and CYNSS surface water extent metrics over the Yucatan Lake wetland site

Wetlands have a major role in the carbon cycle, outgassing large quantities of carbon dioxide and methane through processes that are directly and strongly influenced by the duration and timing of inundation. Therefore, understanding the seasonal pattern of inundation can be a component for regional to global scale carbon models. Measurement of inundation extent also establishes a benchmark for the current status of wetland areas, useful in assessing the future impacts of climate change. The incorporation of frequent measurements of inundation extent into large-scale hydrological models would permit the evaluation of more detailed seasonal and longer-term floodplain dynamics and their associated management implications.The Cyclone GNSS (CYGNSS) constellation of satellites launched in 2016, and carries receivers capable of receiving data from L-band GNSS reflections. Delay Doppler maps (DDM) are generated on board and telemetered to the ground, along with a small number of raw data takes that can be used for special processing on the ground for evaluation purposes. It has been previously shown that these data can be sensitive to inundation.The NASA ISRO Synthetic Aperture Radar, currently planned for launch in January 2023, has both an L-band and S-band SAR for earth imaging. The L-band SAR, which will image the Earth's land mass twice every 12 days, has a requirement for measuring wetland inundation extent at the 1 ha scale. One of the sites that will be used to validate this requirement is Yucatan Lake, Louisiana. This oxbow lake and surrounding area located adjacent to the Mississippi river experiences periodic and extensive flooding in the surrounding forest areas.In 2019, NASA's UAVSAR fully polarimetric airborne L-band SAR conducted a flight campaign to image a dozen sites in the SE USA at approximately 12-day intervals and both in the morning and evening, to simulate the type of data NISAR will obtain. One site imaged during this campaign was the Yucatan Lake area, spanning water stages from low to high flood conditions.It has been demonstrated previously that GNSS reflectometry such as that measured by CYGNSS may be used to characterize surface inundation. It has also been known for decades that L-band SAR may be used to characterize not only the presence of open water, but also the presence of subcanopy inundation in forested areas. In this paper we will present results comparing data from these two types of instruments.

Lavalle, Marco↗

NASA NTRSDate not supplied

Biomass Harmonization and SAR Analysis with the Multi-mission Algorithm and Analysis Platform (MAAP)

The Multi‐mission Algorithm and Analysis Platform (MAAP) is a collaborative effort between NASA and the European Space Agency (ESA) to support above ground biomass (AGB) research in an open science framework. MAAP brings together relevant data, algorithms, and computing capabilities in a common cloud environment to address the challenges of sharing and processing data from field, airborne and satellite measurements. MAAP was publicly released in October 2021, providing computing capabilities co-located with the data, a collaborative coding and analysis environment, and a set of interoperable tools and algorithms developed to support the estimation and visualization of data. MAAP has allowed scientists from both North America and Europe to collaborate on the generation and analysis/visualization of data derived from multiple, discipline-adjacent missions in an open, collaborative environment that has reached beyond traditional scientific investigation. MAAP has been used to support multiple scientific activities. To date, existing LiDAR data from multiple platforms has been calibrated with field measurements and combined for more comprehensive and accurate estimates of above ground biomass AGB; these LiDAR platforms include airborne (e.g. LVIS), the International Space Station (NASA’s Global Ecosystem Dynamics Investigation (GEDI), and satellites (e.g. ICESat-2). The current challenge is to effectively and seamlessly combine the aforementioned LiDAR-based data with new data sources such as P-band RADAR from ESA’s upcoming BIOMASS mission, existing ESA Sentinel-1 C-band SAR, and the 30 PB/yr of high cadence global coverage L-band SAR data from the upcoming NASA-ISRO SAR (NISAR) mission. Recent analysis using MAAP merged ICESat-2 and optical data (Harmonized Landsat Sentinel) produced the most comprehensively precise estimate of boreal-wide AGB to date. Another effort using MAAP is the production and open distribution of global comparisons of AGB map estimates, including from ICESat-2 and GEDI, to bolster stakeholder uptake for policy applications. These map estimates will feed into the Intergovernmental Panel on Climate Change (IPCC) database, likely aiding the next Global Carbon Stocktake of the UNFCCC. Furthermore, the biomass retrieval intercomparison exercise BRIX-2 could benefit from the MAAP providing standardized test cases (based on airborne campaign and spaceborne data) allowing the community to develop and apply retrieval algorithms based on these test cases, while forthcoming SAR data training curricula could also use the MAAP as a teaching and learning platform. The MAAP is meeting the challenges inherent in international, open science collaboration and large scale computing with a platform that is entirely open source and cloud native, using open standards for data access, manipulation, protocols, and formats. The MAAP data system consists of a dedicated data store whose data is indexed in an online catalog conforming to established metadata, application programmatic interfaces (APIs), and service interface standards, using an implementation of the open sourced NASA Common Metadata Repository. Federation of user identities allows users from either NASA or ESA to access and consume services from the other using a unified metadata catalog for the data utilized across the ESA and NASA MAAP platforms. Similarly, we are exploring how to increase interoperability to achieve a common approach to packaging, orchestrating and executing algorithms, with interoperable access to data for subsetting, fast browse, and cloud-optimized access, all using interoperable standards such as those from the Open Geospatial Consortium (OGC). Designed for interoperability, ESA and NASA utilize a common architecture for the software platform. It provides a cloud-based algorithm development environment (ADE) that enables scientists to develop algorithms collaboratively with access to the MAAP data catalog as well as other data archives. MAAP provides an Eclipse Che-based ADE supporting both Python and R languages, popular in this biomass community. Algorithms developed and containerized within the ADE can be deployed to run to thousands of computational nodes in the MAAP’s data processing system (DPS), dramatically speeding up processing and giving scientists a rapid, iterative turnaround of results. NASA’s implementation of the DPS is based on the Hybrid Science Data System (HySDS) framework, used by NASA flight projects to produce Earth science standard products.

cloud computing↗

DOE OSTI2023

Data-Driven Whole-Genome Clustering to Detect Geospatial, Temporal, and Functional Trends in SARS-CoV-2 Evolution

Current methods for defining SARS-CoV-2 lineages ignore the vast majority of the SARS-CoV-2 genome. We develop and apply an exhaustive vector comparison method that directly compares all known SARS-CoV-2 genome sequences to produce novel lineage classifications. We utilize data-driven models that (i) accurately capture the complex interactions across the set of all known SARS-CoV-2 genomes, (ii) scale to leadership-class computing systems, and (iii) enable tracking how such strains evolve geospatially over time. We show that during the height of the original Omicron surge, countries across Europe, Asia, and the Americas had a spatially asynchronous distribution of Omicron sub-strains. Moreover, neighboring countries were often dominated by either different clusters of the same variant or different variants altogether throughout the pandemic. Analyses of this kind may suggest a different pattern of epidemiological risk than was understood from conventional data, as well as produce actionable insights and transform our ability to prepare for and respond to current and future biological threats.

Jacobson, Daniel↗

DOE OSTI2021

Molecular basis for higher affinity of SARS-CoV-2 spike RBD for human ACE2 receptor

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused substantially more infections, deaths, and economic disruptions than the 2002-2003 SARSCoV. The key to understanding SARS-CoV-2's higher infectivity lies partly in its host receptor recognition mechanism. Experiments show that the human angiotensin converting enzyme 2 (ACE2) protein, which serves as the primary receptor for both CoVs, binds to the receptor binding domain (RBD) of CoV-2's spike protein stronger than SARS-CoV's spike RBD. The molecular basis for this difference in binding affinity, however, remains unexplained from X-ray structures. To go beyond insights gained from X-ray structures and investigate the role of thermal fluctuations in structure, we employ all-atom molecular dynamics simulations. Microseconds-long simulations reveal that while CoV and CoV-2 spike-ACE2 interfaces have similar conformational binding modes, CoV-2 spike interacts with ACE2 via a larger combinatorics of polar contacts, and on average, makes 45% more polar contacts. Correlation analysis and thermodynamic calculations indicate that these differences in the density and dynamics of polar contacts arise from differences in spatial arrangements of interfacial residues, and dynamical coupling between interfacial and non-interfacial residues. Furthermore, these results recommend that ongoing efforts to design spike-ACE2 peptide blockers will benefit from incorporating dynamical information as well as allosteric coupling effects.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Seaweed-derived fucoidans and rhamnan sulfates serve as potent anti-SARS-CoV-2 agents with potential for prophylaxis

Seaweeds represent a rich source of sulfated polysaccharides with similarity to heparan sulfate, a facilitator of myriad virus host cell attachment. For this reason, attention has been drawn to their antiviral activity, including the potential for anti-SARS-CoV-2 activity. We have identified and structurally characterized several fucoidan extracts, including those from different species of brown macroalga, and a rhamnan sulfate from a green macroalga species. A high molecular weight fucoidan extracted from Saccharina japonica (FSjRPI-27), and a rhamnan sulfate extracted from Monostroma nitidum (RSMn), showed potent competitive inhibition of spike glycoprotein receptor binding to a heparin-coated SPR chip. This inhibition was also observed in cell-based assays using hACE2 HEK-293 T cells infected by pseudotyped SARS-CoV-2 virus with IC50 values <1 μg/mL. Effectiveness was demonstrated in vivo using hACE2–transgenic mice. Intranasal administration of FSjRPI-27 showed protection when dosed 6 h prior to and at infection, and then every 2 days post-infection, with 100 % survival and no toxicity at 104 plaque-forming units per mouse vs. buffer control. At 5-fold higher virus dose, FSjRPI-27 reduced mortality and yielded reduced viral titers in bronchioalveolar fluid and lung homogenates vs. buffer control. Furthermore, these findings suggest the potential application of seaweed-based sulfated polysaccharides as promising anti-SARS-CoV-2 prophylactics.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2025

Integrated lipidomic and proteomic profiling reveals metabolic network disruption by SARS-CoV-2 variants

The rapid evolution of SARS-CoV-2 has produced myriad viral strains with increasing transmissibility and capacity for immune evasion. While effective vaccination campaigns have reduced the fatalities associated with SARS-CoV-2, infections continue, and a detailed understanding of how this virus manipulates host biochemical pathways remains elusive. We asked both whether the patterns of host lipid rewiring remained consistent across variants and whether the changes in the abundance of lipid classes are related to changes in the expression of the enzymes involved in their biosynthesis. We compared global nontargeted lipidomics on A549-ACE2 cells infected with the delta variant (B.1.617.2), or the omicron (B.1.1.529) variant to our previous results of global nontargeted lipidomics on A549-ACE2 cells infected with the original WA1 strain and further performed quantitative proteomics to assess changes in the host proteome. We found that metabolic rewiring, both on the lipid and the enzymatic level, is remarkably consistent across all three variants. We further mapped changes in the expression of host metabolic enzymes, linking enzyme expression to alterations in the abundance of specific lipids during infection. This analysis identified key proteins related to virus-mediated changes in lipid abundance, including fatty acid synthase (FASN), lysosomal acid lipase (LIPA), and ORMDL, a regulator of sphingolipid biosynthesis. These integrated lipidomic and proteomic experiments shed light on the importance of the complex network of host metabolism networks that support SARS-CoV-2 infection and suggest that lipid metabolism may be a promising avenue for uncovering conserved therapeutic targets.

SARS-CoV-2↗

DOE OSTI2021

Circulating SARS-CoV-2 spike N439K variants maintain fitness while evading antibody-mediated immunity

SARS-CoV-2 can mutate and evade immunity, with consequences for efficacy of emerging vaccines and antibody therapeutics. Here, we demonstrate that the immunodominant SARS-CoV-2 spike (S) receptor binding motif (RBM) is a highly variable region of S and provide epidemiological, clinical, and molecular characterization of a prevalent, sentinel RBM mutation, N439K. We demonstrate N439K S protein has enhanced binding affinity to the hACE2 receptor, and N439K viruses have similar in vitro replication fitness and cause infections with similar clinical outcomes as compared to wild type. We show the N439K mutation confers resistance against several neutralizing monoclonal antibodies, including one authorized for emergency use by the US Food and Drug Administration (FDA), and reduces the activity of some polyclonal sera from persons recovered from infection. Immune evasion mutations that maintain virulence and fitness such as N439K can emerge within SARS-CoV-2 S, highlighting the need for ongoing molecular surveillance to guide development and usage of vaccines and therapeutics.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2023

Modular adjuvant-free pan-HLA-DR-immunotargeting subunit vaccine against SARS-CoV-2 elicits broad sarbecovirus-neutralizing antibody responses

Subunit vaccines typically require co-administration with an adjuvant to elicit protective immunity, adding development hurdles that can impede rapid pandemic responses. To circumvent the need for adjuvant in a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) subunit vaccine, we engineer a thermostable immunotargeting vaccine (ITV) that leverages the pan-HLA-DR monoclonal antibody 44H10 to deliver the viral spike protein receptor-binding domain (RBD) to antigen-presenting cells. X-ray crystallography shows that 44H10 binds to a conserved epitope on HLA-DR, providing the basis for its broad HLA-DR reactivity. Adjuvant-free ITV immunization in rabbits and ferrets induces robust anti-RBD antibody responses that neutralize SARS-CoV-2 variants of concern and protect recipients from SARS-CoV-2 challenge. We demonstrate that the modular nature of the ITV scaffold with respect to helper T cell epitopes and diverse RBD antigens facilitates broad sarbecovirus neutralization. Our findings support anti-HLA-DR immunotargeting as an effective means to induce strong antibody responses to subunit antigens without requiring an adjuvant.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2022

A systematic exploration of boceprevir-based main protease inhibitors as SARS-CoV-2 antivirals

Boceprevir is an HCV NSP3 inhibitor that was explored as a repurposed drug for COVID-19. It inhibits the SARS-CoV-2 main protease (M Pro ) and contains an α-ketoamide warhead, a P1 β-cyclobutylalanyl moiety, a P2 dimethylcyclopropylproline, a P3 tert-butylglycine, and a P4 N-terminal tert-butylcarbamide. By introducing modifications at all four positions, we synthesized 20 boceprevir-based M Pro inhibitors including PF-07321332 and characterized their M Pro inhibition potency in test tubes (in vitro) and 293T cells (in cellulo). Crystal structures of M Pro bound with 10 inhibitors and cytotoxicity and antiviral potency of 4 inhibitors were characterized as well. Replacing the P1 site with a β-(S-2-oxopyrrolidin-3-yl)-alanyl (Opal) residue and the warhead with an aldehyde leads to high in vitro potency. The original moieties at P2, P3 and the P4 N-terminal cap positions in boceprevir are better than other tested chemical moieties for high in vitro potency. In crystal structures, all inhibitors form a covalent adduct with the M Pro active site cysteine. Here, the P1 Opal residue, P2 dimethylcyclopropylproline and P4 N-terminal tert-butylcarbamide make strong hydrophobic interactions with M Pro , explaining high in vitro potency of inhibitors that contain these moieties. A unique observation was made with an inhibitor that contains a P4 N-terminal isovaleramide. In its M Pro complex structure, the P4 N-terminal isovaleramide is tucked deep in a small pocket of M Pro that originally recognizes a P4 alanine side chain in a substrate. Although all inhibitors show high in vitro potency, they have drastically different in cellulo potency to inhibit ectopically expressed M Pro in human 293T cells. In general, inhibitors with a P4 N-terminal carbamide or amide have low in cellulo potency. This trend is reversed when the P4 N-terminal cap is changed to a carbamate. The installation of a P3 O-tert-butyl-threonine improves in cellulo potency. Three molecules that contain a P4 N-terminal carbamate were advanced to cytotoxicity tests on 293T cells and antiviral potency tests on three SARS-CoV-2 variants. They all have relatively low cytotoxicity and high antiviral potency with EC 50 values around 1 μM. A control compound with a nitrile warhead and a P4 N-terminal amide has undetectable antiviral potency. Based on all observations, we conclude that a P4 N-terminal carbamate in a boceprevir derivative is key for high antiviral potency against SARS-CoV-2.

60 APPLIED LIFE SCIENCES↗