Periodic Reporting for period 1 - SupraVir (Multivalent Supramolecular Nanosystems as Dynamic Virus Blockers)
Berichtszeitraum: 2022-10-01 bis 2025-03-31
In the SupraVir project, we proposed multivalent supramolecular concept to specifically target viruses through electrostatic and glycol-interactions. Supramolecular structures involve the self-assembly of molecules through noncovalent interactions, offer several advantages, e.g. flexibility and adaptivity for virus inhibition. Furthermore, in this project, additional functionalities integrated into the structures to enhance the effectiveness of the designed systems in virus inhibition. Several studies have demonstrated the potential of multivalent systems as effective tools for virus inhibition. Therefore, in this project, polymeric supramolecular structures with receptors like sialic acid or sulfate groups are designed to enhance virus-inhibition efficiency. Such an inhibitory effect is expected through electrostatic interactions between the functional groups and spike protein of viruses to block cell internalization of viruses. This concept highlights the potential of polymeric multivalent supramolecular systems, in the field of virus inhibition in order to provide better tools for future virus pandemics.
By far, we have successfully developed self-assembled multivalent structures in three different work packages (WP1-3). In the first WP, we focused on the synthesis and fabrication of benzotriamide (BTA) co-assemblies with functional surfactant motifs which self-assemble into 1D structures. Furthermore, a cellulose based self-assembled 1D nanofiber was developed. In the second WP, we synthesized multivalent 2D polymers based on dendritic amphiphiles which assemble into sheet like aggregates. In WP3, we synthesized 3D self-assemblies in the form of micelles, polymersomes and nanogels. All the synthesized structures have been functionalized with receptor groups like sialic acid or charged sulfate/carboxylic acid groups to increase their interactions with virus particles. The synthesized structures are fully characterized by spectroscopic and microscopic techniques. Additionally in this project, we studied the effect of different substitutions of the self-assembly behavior of final systems and their physicochemical properties.
The evaluations of the SupraVir structures were performed in WP4-5. The structures have been investigated with various biophysical and biological methods. Microscale thermophoresis (MST) was used to investigate the virus binding quantitatively, and cryo-EM was used to visualize virus interactions as WP4. In WP5, for the virus inhibition, they were evaluated by plaque reduction assays, virus infection assays, virucidal assays against multiple viruses, including herpes simplex virus (HSV), influenza, respiratory syncytial virus (RSV) and SARS-CoV-2 etc. 3D in vitro models, such “lung-on-a-chip” models, are being developed for further evaluations.
The evaluations of the SupraVir structures were performed in WP4-5. The structures have been investigated with various biophysical and biological methods. Microscale thermophoresis (MST) was used to investigate the virus binding quantitatively, and cryo-EM was used to visualize virus interactions as WP4. In WP5, for the virus inhibition, they were evaluated by plaque reduction assays, virus infection assays, virucidal assays against multiple viruses, including herpes simplex virus (HSV), influenza, respiratory syncytial virus (RSV) and SARS-CoV-2 etc. 3D in vitro models, such “lung-on-a-chip” models, are being developed for further evaluations.
In WP1, we disclosed the amplification of asymmetry of BTA helical supramolecular polymers by co-assembly with homochiral nonionic surfactants. For these mixtures, a strong amplification of asymmetry was observed by translating the molecular chirality of the surfactant into a preferred helicity of the co-assembled polymers. Using a combination of spectroscopy and microscopy, we found that surfactants change fiber morphology from racemic double helices to single helices with a preferred handedness.
In the investigation of 2D self-assembly nanosheets, we successfully demonstrated that these self-assembled nanosheets can wrap whole virus particles, leading to irreversible virus interactions. Unlike the state-of-the-art multivalent inhibitors, which primarily target the spike protein of the virus, the 2D supramolecular nanosheets introduce a novel approach to permanently inactivate viruses through a virucidal mechanism. Interestingly, the same 2D nanosheet was also effective in inhibiting the infection of influenza A virus, which utilizes a binding ligand distinct from that of SARS-CoV-2. This finding validates our hypothesis that flexible and dynamic supramolecular structures can adapt to varying virus morphologies, offering broad-spectrum antiviral activity. Furthermore, the nanosheets' low IC50 value (30 nM) not only supports their potential for translational studies, but also inspires further exploration of supramolecular nanostructures for diverse biomedical applications.
In the investigation of 2D self-assembly nanosheets, we successfully demonstrated that these self-assembled nanosheets can wrap whole virus particles, leading to irreversible virus interactions. Unlike the state-of-the-art multivalent inhibitors, which primarily target the spike protein of the virus, the 2D supramolecular nanosheets introduce a novel approach to permanently inactivate viruses through a virucidal mechanism. Interestingly, the same 2D nanosheet was also effective in inhibiting the infection of influenza A virus, which utilizes a binding ligand distinct from that of SARS-CoV-2. This finding validates our hypothesis that flexible and dynamic supramolecular structures can adapt to varying virus morphologies, offering broad-spectrum antiviral activity. Furthermore, the nanosheets' low IC50 value (30 nM) not only supports their potential for translational studies, but also inspires further exploration of supramolecular nanostructures for diverse biomedical applications.