Coronavirus disease 2019 (COVID-19) is caused by a new, highly pathogenic severe-acute-respiratory syndrome coronavirus 2 (SARS-CoV-2) that infects human cells through its transmembrane spike (S) glycoprotein. The receptor-binding domain (RBD) of the S protein interacts with the angiotensin-converting enzyme II (ACE2) receptor of the host cells. Therefore, pharmacological targeting of this interaction might prevent infection or spread of the virus. Here, we performed a virtual screening to identify small molecules that block S-ACE2 interaction. Large compound libraries were filtered for drug-like properties, promiscuity and protein-protein interaction-targeting ability based on their ADME-Tox descriptors and also to exclude pan-assay interfering compounds. A properly designed AI-based virtual screening pipeline was applied to the remaining compounds, comprising approximately 10% of the starting data sets, searching for molecules that could bind to the RBD of the S protein. All molecules were sorted according to their screening score, grouped based on their structure and postfiltered for possible interaction patterns with the ACE2 receptor, yielding 31 hits. These hit molecules were further tested for their inhibitory effect on Spike RBD/ACE2 (19-615) interaction. Six compounds inhibited the S-ACE2 interaction in a dose-dependent manner while two of them also prevented infection of human cells from a pseudotyped virus whose entry is mediated by the S protein of SARS-CoV-2. Of the two compounds, the benzimidazole derivative CKP-22 protected Vero E6 cells from infection with SARS-CoV-2, as well. Subsequent, hit-to-lead optimization of CKP-22 was effected through the synthesis of 29 new derivatives of which compound CKP-25 suppressed the Spike RBD/ACE2 (19-615) interaction, reduced the cytopathic effect of SARS-CoV-2 in Vero E6 cells (IC₅₀ = 3.5 μM) and reduced the viral load in cell culture supernatants. Early in vitro ADME-Tox studies showed that CKP-25 does not possess biodegradation or liver metabolism issues, while isozyme-specific CYP450 experiments revealed that CKP-25 was a weak inhibitor of the CYP450 system. Moreover, CKP-25 does not elicit mutagenic effect on Escherichia coli WP2 uvrA strain. Thus, CKP-25 is considered a lead compound against COVID-19 infection.

The NerveRepack project is a European initiative that aims to develop biomimetic exoskeletons/exoprostheses for amputated or paralyzed leg patients that will receive and transmit signals to enable movements and sensations for the patient. To implement the project, it is fundamental to develop implantable neuronal electrodes that will allow bidirectional signaling between the sensors placed on the exoskeletons/exoprostheses and the nervous system. In this direction, two electrodes, plug and cuff, have been designed as integral parts of the final implantable device. The electrodes should comply with strict regulations to ensure their safe implantation in patients. The purpose of this study was to support the compliance of the implant platforms of certain key components with the ISO and ASTM standards that would be required for clinical applications. We have used an indirect method to assess the biocompatibility of the developed electrodes against neuronal cells, fibroblasts, and keratinocytes. Also, we assessed hemocompatibility, i.e., the potential of implantable electrodes to induce hemolysis or complement activation. Finally, the mutagenic/genotoxic potential was tested against the internationally recommended CHO cells. Both representative plug and cuff electrode components were found non-cytotoxic, non-mutagenic, and unable to induce hemolysis. Therefore, from the point of early evaluation of in vitro material and process biocompatibility, the selected implant platforms for the electrodes could be implanted in preclinical models to delineate their potential in vivo applications as neuronal interface with the biomimetic exoskeleton/exoprostheses.