Screening of small molecules to fight SARS-CoV-2 by combining novel approaches and sustainability

Project Information

SPOTLESS

Grant agreement ID: 101024974

DOI

10.3030/101024974

Project closed

EC signature date 22 March 2021

Start date 1 May 2021

End date 30 April 2024

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 257 209,92

EU contribution

€ 257 209,92

Coordinated by

UNIVERSITA DEGLI STUDI DI PADOVA
Italy

Periodic Reporting for period 1 - SPOTLESS (Screening of small molecules to fight SARS-CoV-2 by combining novel approaches and sustainability)

Reporting period: 2021-05-01 to 2024-04-30

Summary of the context and overall objectives of the project

This project addresses the urgent need for effective antiviral treatments against SARS-CoV-2, the virus causing COVID-19. The virus's rapid spread and high mortality rate, along with the limited effectiveness of existing treatments, highlight the necessity for new therapeutic strategies. The project focuses on the viral spike (S) protein, crucial for the virus's entry and infection of human cells. The spike protein is cleaved by proprotein convertases (PCs), a step essential for viral infectivity. Inhibiting this cleavage could significantly reduce the virus's ability to infect and spread. Since the convertase Furin also plays a critical role in other severe diseases, including cancer and deadly infections, finding novel inhibitors is highly motivated.
This research is vital for society, given the global health and economic crises caused by COVID-19. Developing effective antiviral drugs is crucial for managing the current pandemic and preparing for future viral outbreaks. By targeting PCs and developing innovative high-throughput screening (HTS) platforms, the project aims to identify potent inhibitors for new antiviral drugs. These drugs could be essential tools in the global fight against COVID-19 and other viral diseases, ultimately saving lives and helping restore normalcy to society.

Objectives

The project aimed to thoroughly understand the interaction between the SARS-CoV-2 spike protein and proprotein convertases, particularly Furin. It sought to develop two high-throughput screening platforms: an in vitro platform using synthetic substrates and soluble enzymes, and a cell-based platform using luciferase and apoptosis sensors. Additionally, the project emphasized sustainability in research by minimizing the use of non-renewable resources, recycling laboratory materials, and promoting eco-friendly practices.

Achievements

The project successfully advanced the understanding of how the SARS-CoV-2 spike protein interacts with proprotein convertases. It developed innovative platforms for screening potential inhibitors, leading to the identification of several promising compounds that could serve as leads for new antiviral drugs. The research provided detailed insights into the cleavage of the spike protein by Furin, a process essential for the virus's ability to infect human cells. Two high-throughput screening platforms were created: an in vitro platform using synthetic substrates and soluble enzymes, and a novel cell-based platform using luciferase and apoptosis sensors.

The project also identified several small molecules that effectively inhibit Furin activity, showing promise in reducing the infectivity of SARS-CoV-2. Sustainable research practices were incorporated throughout the project, including minimizing the use of non-renewable resources, recycling materials, and promoting eco-friendly practices.

In summary, this project addressed a critical need in the fight against COVID-19 by targeting a novel aspect of the virus's life cycle, with significant implications for public health and future pandemic preparedness. It not only contributed to the scientific understanding of SARS-CoV-2 but also provided practical tools and potential therapeutic leads to combat the virus and other PC-related pathologies, such as cancer and other viral infections.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

This project aimed to understand SARS-CoV-2 and develop potential treatments while emphasizing sustainability and broad dissemination of results.
Significant strides were made in analyzing the SARS-CoV-2 spike protein. Different versions of the spike protein were identified, and it was discovered how enzymes break them down. The spike protein is cut initially quickly, then more slowly, which may help control the virus's activity. Certain combinations of protein building blocks enhance the enzyme Furin's efficiency in cutting the spike protein. A major breakthrough was the development of a new test substrate, QTQTKSHRRAR-AMC, which outperformed older substrates in enzyme tests. This substrate enabled high-throughput screening (HTS), identifying strong Furin inhibitors like the compound P3, which blocks Furin specifically targeting viral proteins without affecting other functions. A new testing system, the SARS-GLuc sensor system, was also created to identify substances that can block SARS-CoV-2 in cells.
The project implemented plastic reduction and recycling, energy-saving practices, and the use of environmentally friendly chemicals. Preferred suppliers with good environmental practices were chosen.
Significant progress was made in understanding SARS-CoV-2 and developing new treatment methods. High sustainability standards were set, and findings were widely shared, contributing valuable knowledge to the scientific community and public health efforts.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

This project has advanced the understanding of the interaction between the SARS-CoV-2 spike protein and the enzyme Furin, which is crucial for the virus entering host cells. Key achievements include:
Detailed Mechanistic Insights: The project has provided in-depth insights into how the spike protein is cleaved by Furin, detailing the specific amino acid sequences and structural changes involved.
Innovative High-Throughput Screening (HTS) Platforms: The development of both in vitro and cell-based HTS platforms has allowed for efficient identification of small molecule inhibitors that block Furin's activity, accelerating the discovery of potential antiviral drugs.
Identification of Promising Inhibitors: Several small molecules have been identified that inhibit Furin activity and reduce the infectivity of SARS-CoV-2 in cellular models, paving the way for effective antiviral therapies.
These advancements not only enhance the understanding of SARS-CoV-2 but also have broader implications for developing antiviral drugs targeting Furin, which could apply to other pathogens as well.

Expected Results Until the End of the Project
Validation of Inhibitors: Further validation of Furin inhibitors in various biological models to confirm their efficacy.
Optimization of Drug Candidates: Refinement of lead inhibitors to improve their potency and specificity.
Expansion of HTS Applications: Applying HTS platforms to other viral pathogens, broadening the research's impact.

Potential Impacts
Reduced Healthcare Burden: Effective antiviral treatments could lower severe COVID-19 cases, easing healthcare system strain and reducing costs.
Economic Recovery: Controlling COVID-19 spread and severity would support faster economic recovery.
Job Creation: Advancements in drug development and new pharmaceutical processes could create jobs in biotechnology and healthcare sectors.

Wider Societal Implications
Public Health Preparedness: The research provides a framework for quickly developing antiviral treatments against future viral threats.
Global Health Equity: Effective, affordable antiviral treatments could benefit low- and middle-income countries with limited access to vaccines.
Scientific Advancement: The project advances understanding of viral mechanisms and host-pathogen interactions, paving the way for future research and innovations.
In summary, this project significantly advances scientific knowledge and technological capabilities in fighting COVID-19, with potential socio-economic benefits and improvements in global health outcomes.

Screening platforms to find SARS-CoV-2 inhibitors