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Entry, Descent, and Landing Communications for the 2007 Phoenix Mars Lander

This paper addresses NASA's requirement on the 2007 Phoenix Mars Lander to provide spacecraft communications during entry, descent, and landing on Mars to allow the identification of probable root cause should any mission failure occur. The Phoenix mission launched on 4 August 2007 and will land on 25 May 2008 on the northern plains of Mars to conduct a three-month study of the Martian environment. The paper discusses the architectural trades in designing a communications link and surveys the entry, descent, and landing communications approaches taken by previous missions. It then discusses the Phoenix-specific constraints and degrees of freedoms and presents a novel and robust implementation approach to entry, descent, and landing communications. The overall methodology and conclusions described herein can serve as a pathfinder for the entry, descent, and landing communications architecture and implementation of future Mars landed missions.

communications↗

Entry, Descent, and Landing Communications for the 2011 Mars Science Laboratory

The Mars Science Laboratory (MSL), established as the most advanced rover to land on the surface of Mars to date, launched on November 26th, 2011 and arrived to the Martian Gale Crater during the night of August 5th, 2012 (PDT). MSL will investigate whether the landing region was ever suitable to support carbon-based life, and examine rocks, soil, and the atmosphere with a sophisticated suite of tools. This paper addresses the flight system requirement by which the vehicle transmitted indications of the following events using both X-band tones and UHF telemetry to allow identification of probable root causes should a mission anomaly have occurred: Heat-Rejection System (HRS) venting, completion of the cruise stage separation, turn to entry attitude, atmospheric deceleration, bank angle reversal commanded, parachute deployment, heatshield separation, radar ground acquisition, powered descent initiation, rover separation from the descent stage, and rover release. During Entry, Descent, and Landing (EDL), the flight system transmitted a UHF telemetry stream adequate to determine the state of the spacecraft (including the presence of faults) at 8 kbps initiating from cruise stage separation through at least one minute after positive indication of rover release on the surface of Mars. The flight system also transmitted X-band semaphore tones from Entry to Landing plus one minute although since MSL was occulted, as predicted, by Mars as seen from the Earth, Direct-To-Earth (DTE) communications were interrupted at approximately is approx. 5 min after Entry ( approximately 130 prior to Landing). The primary data return paths were through the Deep Space Network (DSN) for DTE and the existing Mars network of orbiting assets for UHF, which included the Mars Reconnaissance Orbiter (MRO), Mars Odyssey (ODY), and Mars Express (MEX) elements. These orbiters recorded the telemetry data stream and returned it back to Earth via the DSN. The paper also discusses the total power received during EDL and the robustness of the telecom design strategy used to ensure EDL communications coverage.

relay operations↗

Thermophysics Issues Relevant to High-Speed Earth Entry of Large Asteroids

Physics of atmospheric entry of meteoroids was an active area of research at NASA ARC up to the early 1970s (e.g., the oft-cited work of Baldwin and Sheaffer). However, research in the area seems to have ended with the Apollo program, and any ties with an active international meteor physics community seem to have significantly diminished thereafter. In the decades following the 1970s, the focus of entry physics at NASA ARC has been on improvement of the math models of shock-layer physics (especially in chemical kinetics and radiation) and thermal response of ablative materials used for capsule heatshields. With the overarching objectives of understanding energy deposition into the atmosphere and fragmentation, could these modern analysis tools and processes be applied to the problem of atmospheric entry of meteoroids as well? In the presentation we will explore: (i) the physics of atmospheric entries of meteoroids using our current state-of-the-art tools and processes, (ii) how multiple bodies interact, and (iii) the influence of wall blowing on flow dynamics.

Atmospheric Entry↗

Entry at Venus

This is lecture to be given at the IPPW 2016, as part of the 2 day course on Short Course on Destination Venus: Science, Technology and Mission Architectures. The attached presentation material is intended to be introduction to entry aspects of Venus in-situ robotic missions. The presentation introduces the audience to the aerodynamic and aerothermodynamic aspects as well as the loads, both aero and thermal, generated during entry. The course touches upon the system design aspects such as TPS design and both high and low ballistic coefficient entry system concepts that allow the science payload to be protected from the extreme entry environment and yet meet the mission objectives.

Thermal protection↗

ADEPT, A Mechanically Deployable Re-Entry Vehicle System, Enabling Interplanetary CubeSat and Small Satellite Missions

There is growing interest for utilizing Small Satellites beyond low Earth orbit. A number of secondary CubeSat payload missions are planned at Mars, cis-Lunar Space, near Earth objects, and moons of the Gas Giants. Use of smaller systems may enable utilization of otherwise unused capacity of larger "host" missions. Development of re-entry systems that leverage and accommodate Small Satellite technology will substantially expand the range of mission applications by offering the capability for high speed entry or aerocapture at destinations with atmospheres. Deployable entry vehicles (DEVs) offer benefits over traditional rigid aeroshells including volume, mass and payload form factor. The Adaptive Deployable Entry and Placement Technology (ADEPT) offers such a delivery capability for Small Sat or CubeSat orbiter(s), in-situ elements, or landers. The ADEPT system can package with off the shelf CubeSat deployment systems (1U-16U) to offer a delivery capability for a single CubeSat or constellations. Furthermore, ADEPT can deliver the same science payload to a destination with a stowed diameter a factor of 3-4 times smaller than an equivalent rigid aeroshell, alleviating volumetric constraints on the secondary payload accommodation or primary carrier spacecraft bus. This paper will describe ADEPT's current development status and define various interplanetary mission concepts in order to provide guidelines for potential Small Satellite payload developers and mission implementers.

Adaptive Deployable Entry and Placement Technology↗

Overview of Heatshield for Extreme Entry Environment Technology (HEEET)

The objective of the Heatshield for Extreme Entry Environment Technology (HEEET) projects is to mature a 3-D Woven Thermal Protection System (TPS) to Technical Readiness Level (TRL) 6 to support future NASA missions to destinations such as Venus and Saturn. Destinations that have extreme entry environments with heat fluxes > 3500 W/cm2 and pressures up to 5 atmospheres, entry environments that NASA has not flown since Pioneer-Venus and Galileo. The scope of the project is broad and can be split into roughly four areas, Manufacturing/Integration, Structural Testing and Analysis, Thermal Testing and Analysis and Documentation. Manufacturing/Integration covers from raw materials, piece part fabrication to final integration on a 1-meter base diameter 45-degree sphere cone Engineering Test Unit (ETU). A key aspect of the project was to transfer as much of the manufacturing technology to industry in preparation to support future mission infusion. The forming, infusion and machining approaches were transferred to Fiber Materials Inc. and FMI then fabricated the piece parts from which the ETU was manufactured. The base 3D-woven material consists of a dual layer weave with a high-density outer layer to manage recession in the system and a lower density, lower thermal conductivity inner layer to manage the heat load. At the start of the project it was understood that due to weaving limitations the heat shield was going to be manufactured from a series of tiles. And it was recognized that the development of a seam solution that met the structural and thermal requirements of the system was going to be the most challenging aspect of the project. It was also recognized that the seam design would drive the final integration approach and therefore the integration of the ETU was kept in-house within NASA. A final seam concept has been successfully developed and implemented on the ETU. The structural testing and analysis covers from characterization of the different layers of the infused material as functions of weave direction and temperature to sub-component level testing such as 4pt bend testing at sub-ambient and elevated temperature and culminates in testing of the ETU results from which will validate the structural models developed using the element and sub-component level tests. Given the seam has to perform both structurally and aerothermally during entry a novel 4pt bend test fixture was developed allowing articles to be tested while the front surface is heated with a laser. These tests are being utilized to establish the systems structural capability during entry.A broad range of aerothermal tests (arcjet tests) were performed to develop material response models for predicting the required TPS thickness to meet a missions needs and to evaluate failure modes and establish the capability of the system. The final aspect of the project is to develop a comprehensive Design and Data Book such that a future mission will have the information necessary to adopt the technology. This presentation will provide an overview for each of these areas and argue that HEEET has successfully achieved TRL 6.

HEEET↗

In Situ Small Spacecraft Missions Utilizing Heatshield for Extreme Entry Environments Technology

There is considerable interest in utilizing Small Spacecraft beyond low Earth orbit. In November of 2018, successful data relay operations of the MarCO CubeSats during the entry, descent, and landing (EDL) of the Mars InSight mission showed the viability of using CubeSats for interplanetary missions. Additional testament to the promise of Small Spacecraft class rideshare missions is the upcoming Artemis-1 flight test that will launch thirteen 6U CubeSats, as well as the establishment of NASA’s SIMPLEx program which will conduct stand-alone planetary science missions that launch with a primary payload. It is anticipated that continued innovations in Small Spacecraft capabilities combined with small EDL systems will expand the range of potential missions to allow for in situ investigations. Recently, NASA invested in the development of a new, efficient and capable ablative thermal protection system (TPS), utilizing 3-D Weaving. The new dual layer TPS, Heatshield for Extreme Entry Environment Technology (HEEET), is tailorable, scalable, robust, mass efficient and capable of supporting in situ missions across the solar system. Combining the HEEET entry system with innovative Small Spacecraft technology will substantially expand the range of Small Spacecraft mission applications by offering the capability for high speed entry or aerocapture at destinations with atmospheres. This paper will describe the HEEET aeroshell system and highlight various mission concepts including a dual technology demonstration mission that is under development and other concepts to deliver instruments for planetary science.

Small Spacecraft↗

Venus Cloud Layer Investigation: Aeroshells for Entry, Descent and Deployment

Entry, Descent and Deployment (EDD) of aerial platforms at Venus follows similar operational approach as landers. •Limited only by the availability of mass efficient and robust aeroshell (heatshield/TPS) technology. •Heatshield for Extreme Entry Environment Technology (HEEET) at TRL 6 is an enabler of Venus in-situ missions.Lower ballistic coefficient, deployable concept, ADEPT, offers additional options•Low deceleration entry profile•Release of one or more payloads (balloons) from open back of the entry vehicle2

Venus↗

Impact of NASA’s Entry Systems Modeling Project on Planetary Mission Design

Planetary missions continue to grow larger and more complex. Furthermore, the current focus on human exploration of the Moon and Mars, as well as Mars Sample Return(MSR), place increas-ingly stringent requirements on the reliability of the entry, descent, and landing (EDL) system that ensures the safe delivery of payload or crew to their destination. Planetary EDL is an area in which mission designers are critically reliant on modeling and simulation to demonstrate the reliability of the system, as there are no ground facilities that are able to fully test these systems in a flight-relevant environment. NASA’s state-of-the-art modeling and simulation capability must continually evolve to meet the needs of the next generation of planetary EDL. To accomplish this aim, NASA’s Entry Systems Modeling (ESM) Project was formed in 2013and is funded bythe Space Technology Mission Directorate(STMD) and Science Mission Directorate (SMD). ESM is the Agency’s only cross-cutting effort for advancing entry systems modeling and simulation capabilities across a range of technical disciplines and Solar System destinations. ESM is a portfolio project covering a variety of mid-TRL research efforts within four core EDL-related areas of investment: (1) Thermal protection material modeling, (2) Shock layer kinetics and radiation, (3) Aerosciences, and (4) Guidance, navigation, and con-trol. The material modeling group creates detailed material response modelsof thermal protection systems (TPS)from the micro to macro scale, and at the fun-damental and engineering levels. Shock layer kinetics and radiation focuses on radiative heating of space-craft, quantum chemistry and benchmark experiments for validation.Aerosciences is a broad research area that impacts many aspects of entry systems, including parachutes, aerodynamics, and turbulent heating augmentation due to TPS roughness.The guidance, navigation,and control effort under ESM is expanding the capabilities of NASA’s main flight mechanics tool, POST2, for use on high-performance computing architectures and to generalize interoperability with external applications for more detailed end-to-end simula-tion.In addition, several focusedresearch topics have been approvedto augment ESM’s core portfolio. These include efforts for deep post-flight analysis of Mars 2020/MEDLI2 flight data; development ofTPS failure models; improvement of hypersonic wakeflow models; and a recently concluded effort to provide material response models for NuSil-coated PICA heat-shield material. This presentation will discuss each of these investment areas and demonstrate via real mission examples how advances to the state-of-the-art enabled by ESM are directly impacting the missions of today and tomorrow, including InSight, Mars 2020, Mars Sample Return, Orion, and Dragonfly.

Entry Systems Modeling↗

Design and Technology Maturation of the Stratospheric Projectile Experiment of Entry Dynamics

The supersonic and transonic dynamic stability of blunt-body reentry vehicles currently poses large risks in all of NASA’s ongoing entry missions (MSR SRL, MSR EES, and Dragonfly). These projects have allocated millions of dollars to testing and modeling efforts to buy down risk by using the current state-of-the-art (SoA) facilities at NASA’s disposal. While these facilities have heritage in supplying dynamics data to reentry missions, their availability is severely limited – particularly with the high number of concur-rent projects requesting simultaneous testing– and are costly when considering the science density per dollar. None of the current SoA facility methodologies allow the test model to have the dynamics fully develop through a flight relevant free-stream profile and as such require extrapolations with resultant high uncertainties in order to relate the test dynamics to flight expectations. SPEED is a NASA Ames Center Innovation Fund (CIF) project that is developing a highly tailorable and cost-effective test methodology to better assess the dynamic stability of blunt-body reentry vehicles via a stratospheric balloon flight. This is accomplished by dropping a suite of instrumented capsules from a stratospheric balloon to gain a statistically relevant dataset of scaled reentry vehicles in mission relevant free-flight conditions. This presentation will walk through how the test methodology is being implemented specifically for the Mars Sample Return (MSR) Earth Entry System (EES) geometry in an awarded Flight Opportunities Program (FOP) test flight in early CY24. SPEED Application to MSR: SPEED consists of three main mechanical systems: the Drop Platform, the Projectile, and the test Capsule. SPEED is being developed as a set of guidelines and recommendations for how to test with the proposed Concept of Operations (Conops) since the specific design parameters will vary depending on the specific project’s reference trajectory and entry vehicle design. As such, this presentation will walk through the development time-line as shown in Fig. 2. This is meant to serve as a blueprint for further missions as desired. Mechanical and Avionics Design. The SPEED test platform designed for the MSR-EES capsule geometry with nominal entry parameters has the ability to carry 10 Capsules to altitude instrumented with: 1. 3-Axis Accelerometer 2. IMU 3. Gyroscope 4. Magnetometer 5. Pressure Transducer cruciform 6. Uplook and Horizon Cameras To package the avionics/instrumentation suite, the capsule is approximately 1’ in diameter with the Outer Mold Line (OML) centroid-scaled from the full EES design. The internal volume is gutted and custom-shaped to fit the desired instrumentation suite, as well as to allow for the positioning of ballast mass such that the Center of Gravity is analogous to the flight vehicle. All structural components in the Capsule and Projectile are 3D printed, which significantly reduces the cost of each flight unit to around $1500 including all instrumentation, avionics, and structural components. Flight Conops. The test Capsule is accelerated to the desired altitude and Mach number while stowed in the Projectile, a missile-like vehicle consisting of steel ballast in the nose, a low-drag OML, and an Ejection Mechanism to reliably release the Capsule into the free-flow supersonic conditions. For the MSR-EES design, the capsule employs ~3kg of ballast mass at the nose to accelerate the 1.25kg test Capsule to ~Mach 1.7 at 23km altitude. This requires an initial release altitude of 40km, the quoted limit of a 80kg payload by the FOP-contracted balloon provider. Once the Ejection Mechanism avionics detect the proper conditions, the spring-loaded Ejection Mechanism will release and – guided by the sabot – expose the test Capsule to the desired test conditions for ~5 seconds of free-flight in the supersonic/transonic regimes. Dynamics in the subsonic regime will also be captured with the instrumentation suite with post-flight recovery operations aimed at recovering the high-G-load capable SD cards after the planned hard impact landings. Testing and Development: In the few months the SPEED project has worked the development of MSR-EES flight test, the team has performed lab and drone based testing which this presentation will overview. After the first design phase, the team fabricated Engineering Demonstration Units (EDUs) of all subsystems to perform validation testing shown in Fig. 5. After validation was completed on the subsystem level, a drone-drop test was performed at the recreational flight ceiling of 400ft altitude to assess the SPEED systems in a flight environment. Parameters such as in-flight stability, hard impact landing performance, and avionics performance were quantified and qualified. The FY23 CIF will culminate in a helicopter drop test aboard an Air National Guard Blackhawk. This will prepare the team for the CY24 FOP stratospheric balloon flight that should provide the final verification to begin offering the test platform for mission support. Focus of Presentation: This presentation will outline the technology maturation path of the SPEED implementation to the MSR-EES capsule baseline as well as the details regarding the mechanical system, avionics and instrumentation, and flight operations. Note that a complementary presentation is being submitted for a methodology overview of the SPEED test platform, introducing the testing technique and benefits as well as the full application space of the technology.

pitch damping coefficient↗

Development of a Low-Cost, Highly Flexible Re-Entry Platform

With a growing interest in atmospheric entry, descent, and landing research as well as the necessity to empirically test increasingly complicated entry systems, a growing need for a quick and cost-effective method of experimentation is needed. The Hypersonic Configurable Unit Ballistic Experiment is a test and evaluation platform allowing for in-situ research and rapid testing of novel technologies in a re-entry environment. HyCUBE takes the form of a small satellite-sized re-entry vehicle and, with the additional space on board the vehicle, can act as a platform for testing novel guidance and control methodologies.

Aero-Thermodynamics↗

Assessing Huygens Probe Entry, Descent, and Landing at Titan Simulation using Dragonfly Atmosphere Model

Dragonfly is a New Frontiers Program mission that will deliver a rotorcraft to Saturn's moon, Titan. This mission follows Huygens as the previous mission that successfully landed a vehicle on Titan. A flight mechanics simulation of Dragonfly's Entry, Descent, and Landing sequence has been developed using the Program to Optimize Simulated Trajectories II. The simulation incorporates several subsystem models, including aerodynamics, gravity, and mass properties, to fully capture the multi-body six degree of freedom dynamics. Among all the subsystem models that inform the Entry, Descent, and Landing dynamics, the atmosphere model of Titan is a critical component. The atmosphere model characterizes the density, temperature, pressure, and winds that the entry vehicle experiences during the descent. This impacts several aspects of the descent such as the peak heating, aerodynamics, parachute release conditions, the dynamics of the vehicle and parachutes, and the landing ellipse. In the course of developing Dragonfly, an updated model of the Titan atmosphere has been created corresponding to Dragonfly's arrival in the mid-2030s, approximately one Titan year after the Huygens mission successfully landed a probe on Titan. This work leverages previous work done to investigate Huygens EDL sequence to assess the atmosphere model developed for Dragonfly. This is done by utilizing the updated Titan atmosphere model, the Dragonfly atmosphere model, within the Huygens POST2-based flight simulation with the goal of characterizing the differences between the atmospheric models and assessing how the Dragonfly atmosphere model impacts Huygens entry dynamics.

Huygens↗

Dragonfly Mission Entry and Descent Modeling and Simulation Overview

Dragonfly is a New Frontiers Program mission, led by The Johns Hopkins Applied Physics Laboratory, that will deliver a rotorcraft lander to Saturn’s moon, Titan. The focus of this work is to analyze the trajectory of the entry vehicle from cruise stage separation until lander separation. This analysis is done by the NASA Langley Research Center Entry, Descent, and Landing team using the Program to Optimize Trajectories II. This paper provides an overview of the current design and the robustness of the overall entry sequence using a Monte Carlo uncertainty analysis. The work presented in this study includes the updated design, models, and analysis completed since the Dragonfly Entry, Descent, and Landing Mission Preliminary Design Review.

Simulation↗

Assessing Huygens Probe Entry, Descent, and Landing at Titan Simulation using Dragonfly Atmosphere Model

Dragonfly is a New Frontiers Program mission that will deliver a rotorcraft to Saturn's moon, Titan. This mission follows Huygens as the previous mission that successfully landed a vehicle on Titan. A flight mechanics simulation of Dragonfly's Entry, Descent, and Landing sequence has been developed using the Program to Optimize Simulated Trajectories II. The simulation incorporates several subsystem models, including aerodynamics, gravity, and mass properties, to fully capture the multi-body six degree of freedom dynamics. Among all the subsystem models that inform the Entry, Descent, and Landing dynamics, the atmosphere model of Titan is a critical component. The atmosphere model characterizes the density, temperature, pressure, and winds that the entry vehicle experiences during the descent. This impacts several aspects of the descent such as the peak heating, aerodynamics, parachute release conditions, the dynamics of the vehicle and parachutes, and the landing ellipse. In the course of developing Dragonfly, an updated model of the Titan atmosphere has been created corresponding to Dragonfly's arrival in the mid-2030s, approximately one Titan year after the Huygens mission successfully landed a probe on Titan. This work leverages previous work done to investigate Huygens EDL sequence to assess the atmosphere model developed for Dragonfly. This is done by utilizing the updated Titan atmosphere model, the Dragonfly atmosphere model, within the Huygens POST2-based flight simulation with the goal of characterizing the differences between the atmospheric models and assessing how the Dragonfly atmosphere model impacts Huygens entry dynamics.

Entry Descent Landing↗

Dragonfly Mission Entry and Descent Modeling and Simulation Overview

Dragonfly is a New Frontiers Program mission, led by The Johns Hopkins Applied Physics Laboratory, that will deliver a rotorcraft lander to Saturn’s moon, Titan. The focus of this work is to analyze the trajectory of the entry vehicle from cruise stage separation until lander separation. This analysis is done by the NASA Langley Research Center Entry, Descent, and Landing team using the Program to Optimize Trajectories II. This paper provides an overview of the current design and the robustness of the overall entry sequence using a Monte Carlo uncertainty analysis. The work presented in this study includes the updated design, models, and analysis completed since the Dragonfly Entry, Descent, and Landing Mission Preliminary Design Review.

Simulation↗

AI-Enhanced Computational Tools for Entry Systems Modeling

To advance the understanding of complex atmospheric entry phenomena, NASA’s Entry Systems Modeling (ESM) team [1] has developed high-fidelity computational tools addressing multiscale challenges, from material microstructures to full-scale heatshield response. This abstract highlights a subset of ESM tools, focusing on AI integration to enhance workflows and predictive modeling. - PuMA [2] computes effective material properties from high-resolution micro-CT scans, supporting TPS analysis for NASA missions. - TomoSAM [3] automates 3D tomography dataset segmentation for PuMA using the Segment Anything Model, reducing manual effort and improving accuracy. - PATO [4] models porous reactive materials under extreme conditions, with advancements such as unified solvers, mechanical erosion, and TPS coatings for NASA missions. - arcjetCV [5] employs deep learning to analyze arc jet test footage, measuring recession rates, shape changes, and shock standoff distances, bridging simulations, and experiments to reveal TPS ablation behavior. - ARCHeS [6] simulates arc heater plasma flows, modeling turbulence, radiation, and electromagnetic interactions to optimize arc heater performance, validate TPS under extreme conditions, and serve as a foundation for developing digital twins of arc heater facilities. - SPARTA [7] simulates rarefied hypersonic flows and gas-surface interactions for planetary entry missions, leveraging GPU architectures for scalable and efficient aerothermal and ablation analyses. AI-driven solutions, such as deep learning segmentation, have streamlined workflows in ESM tools and still hold significant potential to further accelerate processes and enhance automation in entry systems modeling. [1] Haskins, J.B. (2023), [2] Ferguson, J.C. (2018), [3] Meurisse, J.B.E. (2018), [4] Semeraro, F. (2023), [5] Quintart, A. (2024) [6] Meurisse, J.B.E. (2022), [7] Plimpton, S.J. (2019)

Predictive Modeling↗

A Novel Device for True Lumen Re-Entry After Subintimal Recanalization of Superficial Femoral Arteries: First-in-Man Experience and Technical Description

Subintimal angioplasty (SAP) is frequently performed for the treatment of critical limb ischemia (CLI) and has been recognized as an effective technique for these patients. Nevertheless, this approach is limited by the lack of controlled re-entry into the true lumen of the target vessel. We describe a novel device for true lumen re-entry after subintimal recanalization of superficial femoral arteries (SFA). We report our experience with six patients treated between April 2009 and January 2010 with a novel system designed to facilitate true lumen re-entry. The device was advanced by ipsilateral antegrade approach through a 6-French sheath. Successful reaccess into the true lumen was obtained in five of six patients without complications. The patient in whom the reaccess to the true lumen was not possible underwent successful bypass surgery. At 30 days follow-up, the SFA was patent in all patients according to echo-Doppler examination. Our preliminary experience indicates that this novel re-entry device increases the success rate of percutaneous revascularization of chronically occluded SFA.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

A new strategy to identify hepatitis B virus entry inhibitors by AlphaScreen technology targeting the envelope-receptor interaction

Highlights: • AlphaScreen evaluating LHBs-NTCP binding identified rapamycin as an anti-HBV agent. • Rapamycin directly interacted with NTCP. • Rapamycin inhibited the infection of HBV and HDV. • Everolimus and temsirolimus possessed higher anti-HBV potency than rapamycin. • First report applying AlphaScreen to the identification of viral entry inhibitors. Current anti-hepatitis B virus (HBV) agents have limited effect in curing HBV infection, and thus novel anti-HBV agents with different modes of action are in demand. In this study, we applied AlphaScreen assay to high-throughput screening of small molecules inhibiting the interaction between HBV large surface antigen (LHBs) and the HBV entry receptor, sodium taurocholate cotransporting polypeptide (NTCP). From the chemical screening, we identified that rapamycin, an immunosuppressant, strongly inhibited the LHBs-NTCP interaction. Rapamycin inhibited hepatocyte infection with HBV without significant cytotoxicity. This activity was due to impaired attachment of the LHBs preS1 domain to cell surface. Pretreatment of target cells with rapamycin remarkably reduced their susceptibility to preS1 attachment, while rapamycin pretreatment to preS1 did not affect its attachment activity, suggesting that rapamycin targets the host side. In support of this, a surface plasmon resonance analysis showed a direct interaction of rapamycin with NTCP. Consistently, rapamycin also prevented hepatitis D virus infection, whose entry into cells is also mediated by NTCP. We also identified two rapamycin derivatives, everolimus and temsirolimus, which possessed higher anti-HBV potencies than rapamycin. Thus, this is the first report for application of AlphaScreen technology that monitors a viral envelope-receptor interaction to identify viral entry inhibitors.

60 APPLIED LIFE SCIENCES↗