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

Wafaa Alabsi; Maria F Acosta; Fahad A Al-Obeidi; Meredith Hay; Robin Polt; Heidi M Mansour

doi:10.3390/pharmaceutics13081278

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

Pharmaceutics. 2021 Aug 17;13(8):1278. doi: 10.3390/pharmaceutics13081278.

Authors

Wafaa Alabsi^{1

2}, Maria F Acosta², Fahad A Al-Obeidi¹, Meredith Hay^{3

4

5}, Robin Polt^{1

2

3}, Heidi M Mansour^{2

3

6}

Affiliations

¹ Department of Chemistry & Biochemistry, The University of Arizona, Tucson, AZ 85721, USA.
² Skaggs Pharmaceutical Sciences Center, College of Pharmacy, The University of Arizona, Tucson, AZ 85721, USA.
³ The BIO5 Institute, The University of Arizona, Tucson, AZ 85721, USA.
⁴ Department of Physiology, The University of Arizona, Tucson, AZ 85721, USA.
⁵ Evelyn F. McKnight Brain Institute, The University of Arizona, Tucson, AZ 85721, USA.
⁶ Division of Translational & Regenerative Medicine, College of Medicine, The University of Arizona, Tucson, AZ 85721, USA.

Abstract

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_g) consistent with the amorphous glassy state formation with T_gs 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.

Keywords: ALI; SPPS; TEER; air–liquid interface; angiotensin; brain; cell viability; glycopeptide; in vitro 2D/3D cell culture; partitioning; solid phase peptide synthesis; solubility; transepithelial electrical resistance.

Abstract

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