Synthesis, Physicochemical Characterization, In Vitro 2D/3D Human Cell Culture, and In Vitro Aerosol Dispersion Performance of Advanced Spray Dried and Co-Spray Dried Angiotensin (1-7) Peptide and PNA5 with Trehalose as Microparticles/Nanoparticles for Targeted Respiratory Delivery as Dry Powder Inhalers

• Published in: Pharmaceutics 2021-08, Vol.13 (8), p.1278
• Main Authors: Alabsi, Wafaa, Acosta, Maria F, Al-Obeidi, Fahad A, Hay, Meredith, Polt, Robin, Mansour, Heidi M
• Format: Article
• Language: English
• Subjects: Aerosols; Amino acids; angiotensin; Antibiotics; Bioavailability; Biodistribution; Blood pressure; brain; Coronaviruses; COVID-19; Disease; Enzymes; glycopeptide; Hypertension; in vitro 2D/3D cell culture; Inflammation; Inhalers; Lungs; Morphology; Particle size; Peptides; Permeability; Phase transitions; Severe acute respiratory syndrome coronavirus 2; solid phase peptide synthesis; SPPS
• ISSN: 1999-4923; 1999-4923
• DOI: 10.3390/pharmaceutics13081278

The peptide hormone Angiotensin (1-7), Ang (1-7) or (Asp-Arg-Val-Tyr-Ile-His-Pro), is an essential component of the renin-angiotensin system (RAS) peripherally and is an agonist of the Mas receptor centrally. Activation of this receptor in the CNS stimulates various biological activities that make the Ang (1-7)/MAS axis a novel therapeutic approach for the treatment of many diseases. The related O-linked glycopeptide, Asp-Arg-Val-Tyr-Ile-His-Ser-(O-β-D-Glc)-amide (PNA5), is a biousian revision of the native peptide hormone Ang (1-7) and shows enhanced stability in vivo and greater levels of brain penetration. We have synthesized the native Ang (1-7) peptide and the glycopeptide, PNA5, and have formulated them for targeted respiratory delivery as inhalable dry powders. Solid phase peptide synthesis (SPPS) successfully produced Ang (1-7) and PNA5. Measurements of solubility and lipophilicity of raw Ang (1-7) and raw PNA5 using experimental and computational approaches confirmed that both the peptide and glycopeptide have high-water solubility and are amphipathic. Advanced organic solution spray drying was used to engineer the particles and produce spray-dried powders (SD) of both the peptide and the glycopeptide, as well as co-spray-dried powders (co-SD) with the non-reducing sugar and pharmaceutical excipient, trehalose. The native peptide, glycopeptide, SD, and co-SD powders were comprehensively characterized, and exhibited distinct glass transitions (T ) consistent with the amorphous glassy state formation with T s that are compatible with use in vivo. The homogeneous particles displayed small sizes in the nanometer size range and low residual water content in the solid-state. Excellent aerosol dispersion performance with a human DPI device was demonstrated. In vitro human cell viability assays showed that Ang (1-7) and PNA5 are biocompatible and safe for different human respiratory and brain cells.