Novel virus-inspired lipid nanoparticles strategies for antiviral therapy and pulmonary drug delivery in respiratory diseases

• Ano de defesa: 2024
• Autor(a) principal: Martins, Yugo Araújo
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: eng
• Instituição de defesa: Biblioteca Digitais de Teses e Dissertações da USP
• Palavras-chave em Português: Brequinar; Bromexina; Covid-19; Doença infecciosa; HSV; Liberação pulmonar; Mucina; Nanopartículas anti-inflamatórias; Nanopartículas antivirais; Nanopartículas virucidas; Nebulização; SARS-CoV-2; Sulfato de heparana; Tratamento pulmonar; VSR
• Link de acesso: https://www.teses.usp.br/teses/disponiveis/60/60137/tde-12062025-183244/

Biomedical nanomaterials designed for therapeutic or diagnostic purposes encounter significant challenges upon administration. Upon contact with biological media, these materials quickly attract proteins, forming a protein corona, which can profoundly affect their stability, toxicity, targeting capabilities, and clearance, potentially compromising their effectiveness. Depending on the application and route of administration, these nanomaterials must navigate complex biological barriers such as the mucus and epithelial barriers following topical, oral or pulmonary delivery. Moreover, for effective treatment, nanomaterials must precisely target specific cells while avoiding numerous off-target cells to minimize adverse effects. The cellular membrane presents another significant barrier, impeding nanoparticles from easily entering the intracellular space. Even if they breach this barrier, nanoparticles must escape endocytic vesicles to release their payload or reach specific organelles. In contrast, viruses, naturally nanosized particles, excel at overcoming these obstacles. To complete their life cycle, viruses traverse complex biological barriers, evade immune detection and clearance, and specifically recognize and invade host cells, replicating and spreading from one cell to another. Due to their sophisticated evolutionary strategies, viruses can overcome various barriers encountered during host infection. Their morphology, surface properties, responsiveness to stimuli, and interactions with cellular receptors contribute to their remarkable abilities. Inspired by the specific designs and functions of viruses and their interactions with biological interfaces, such as the respiratory mucosa and host cells, this research focused on integrating beneficial viral traits into nanomaterials to enhance their functionality. The main objective of this research was to design lipid nanoparticle (LNPs) platforms that emulate viral features, such as interactions with mucus and viral attachment receptors (VARs), to combat viral infections and enhance drug pulmonary delivery. Therefore, this doctoral thesis comprised three main investigations: (1) Development of LNPs stabilized with polyethylene glycol (PEG) derivatives to target mucin-like regions (MLRs) of viruses and encapsulate brequinar (BQ), a dihydroorotate dehydrogenase (DHODH) inhibitor, thereby presenting a platform with dual antiviral mechanism of action. LNPs formulated with d-α-tocopheryl succinate polyethylene glycol 1000 (TS) (TS-LNPs) showed binding affinity to mucins, modulated mucus transport, and inhibited entry of herpes simplex virus 2 (HSV-2), vesicular stomatitis virus expressing coronavirus spike protein (VSV-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Encapsulation of brequinar (BQ) within TS-LNPs (BQ-LNPs) demonstrated sustained drug release, and enhanced antiviral efficacy against SARS-CoV-2 compared to BQ solution; (2) Development of LNPs that mimic the receptor-binding and cleaving activities of viral surface enzymes to enhance transport across the mucus layer, facilitate cellular internalization, and regulate mucus production and the pronounced inflammatory response associated with pulmonary diseases and viral infections. Composed of low molecular weight PEG-coated lipid nanoparticles with bromhexine (BH), nanobromhexine lipid particles (NBL) displayed stability to nebulization, mucoadhesive properties, effective traversal in the dense mucus layer of Calu-3 cultures in an air-liquid interface, and enhanced internalization by the target respiratory epithelial cells. NBL treatment in SARS-CoV-2-infected lung cells led to a 40-fold increase in anti-inflammatory (mucin 1) MUC1 gene expression, a proportional reduction in pro-inflammatory IL-6 expression, and elevated antiinflammatory IL-10 expression, suggesting a potential mechanism to regulate the excessive IL-6 expression triggered by virus infection; (3) Development of LNPs with modified surface that mimic VARs, such as heparan sulfate proteoglycans (HSPG), to create virucidal and broad-spectrum nanocarriers. To gain a deeper understanding of the mechanism of HSPG-mimicking broad-spectrum virucidal compounds, we crafted new biocompatible antiviral nanotaurodeoxylate lipid particles (NTL) and investigated the concept of HSPG mimicry to provide the key nontoxic virucidal action. We show that NTL are broad-spectrum, biocompatible, and virucidal at micromolar concentrations in vitro against various viruses. They are effective against SARS-CoV-2 in a respiratory tissue culture model as well as when administered in mice before and after intranasal respiratory syncytial virus type A (RSV-A) inoculation.