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. 2020 Oct:64:101975.
doi: 10.1016/j.pupt.2020.101975. Epub 2020 Oct 31.

Advanced spray dried proliposomes of amphotericin B lung surfactant-mimic phospholipid microparticles/nanoparticles as dry powder inhalers for targeted pulmonary drug delivery

Alexan I Gomez  1 , Maria F Acosta  2 , Priya Muralidharan  2 , Jason X-J Yuan  3 , Stephen M Black  4 , Don Hayes Jr  5 , Heidi M Mansour  6
Affiliations

Advanced spray dried proliposomes of amphotericin B lung surfactant-mimic phospholipid microparticles/nanoparticles as dry powder inhalers for targeted pulmonary drug delivery

Alexan I Gomez et al. Pulm Pharmacol Ther. 2020 Oct.

Abstract

The purpose of this study was to design, develop and characterize inhalable proliposomal microparticles/nanoparticles of Amphotericin B (AmB) with synthetic phospholipids, dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) which are lung surfactant-mimic phospholipids. Organic solutions of AmB and phospholipids, were co-spray dried using an advanced closed-mode system and a high performance cyclone. Scanning electron microscopy (SEM) was employed to visualize the surface structure, morphology, and particles size. The residual water content of the proliposomes was quantified by Karl Fisher coulometric titration (KFT). Degree of crystallinity/non-crystallinity was measured by X-ray powder diffraction (XRPD). Phase behavior was measured by differential scanning calorimetry. The chemical composition by molecular fingerprinting was established using attenuated total reflectance (ATR)-Fourier-transform infrared (FTIR) spectroscopy. The amount of AmB loaded into the proliposomes was quantified using UV-VIS spectroscopy. The in vitro aerosol dispersion performance was conducted using the Next Generation Impactor (NGI) and the human dry powder inhaler (DPI) (Handihaler®) that is FDA-approved. Different human lung cell lines were employed to demonstrate in vitro safety as a function of dose and formulation. Smooth, spherical microparticles/nanoparticles were formed at medium and high spray drying pump rates and had low residual water content. A characteristic peak in the XRPD diffraction pattern as well as an endotherm in DSC confirmed the presence of the lipid bilayer structure characteristic in the DPPC/DPPG proliposomal systems. Superior in vitro aerosol performance was achieved with engineered microparticles/nanoparticles demonstrating suitability for targeted pulmonary drug delivery as inhalable dry powders. The in vitro cellular studies demonstrated that the formulated proliposomes are safe. These AmB proliposomes can be a better option for targeted treatment of severe pulmonary fungal infections.

Keywords: Co-spray drying particle engineering; Human inhaler device; In vitro aerosol dispersion; In vitro human pulmonary cell viability; Lung surfactant phospholipids; Pulmonary drug delivery.

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Conflict of interest statement

Conflicts of Interest: The authors declare no conflict of interest.

Figures

Figure 1.
Figure 1.
Chemical Structure of: (a) Amphotericin B (AmB); (b) dipalmitoylphosphatidylcholine (DPPC); and (c) dipalmitoylphosphatidylglycerol (DPPG) sodium salt.
Figure 2.
Figure 2.
SEM Micrographs of Raw AmB, SD Particles, and co-SD particles: (a) Raw AmB; (b) SD AmB (med P); (c) SD AmB (high P), (d) co-SD AmB:DPPC/DPPG (low P); (e) co-SD AmB:DPPC/DPPG med P); and (f) co-SD AmB:DPPC/DPPG (high P). Magnifications for all micrographs were 10,000x.
Figure 3.
Figure 3.
SEM Micrographs of Raw AmB, SD Particles, and co-SD particles: (a) Raw AmB; (b) SD AmB (med P); (c) SD AmB (high P), (d) co-SD AmB:DPPC/DPPG (low P); (e) co-SD AmB:DPPC/DPPG (med P); and (f) co-SD AmB:DPPC/DPPG (high P). Magnifications for all micrographs were 20,000x.
Figure 4.
Figure 4.
X-ray Powder Diffractograms of Raw AmB, SD AmB, and co-SD AmB:DPPC/DPPG.
Figure 5.
Figure 5.
Representative DSC Thermograms for: (a) raw AmB; (b) SD AmB (med P); (c) SD AmB (high P); (d) co-SD AmB:DPPC/DPPG (low P); (e) co-SD AmB:DPPC/DPPG (med P); and (f) co-SD AmB:DPPC/DPPG (high P).
Figure 6.
Figure 6.
Representative HSM Images for: (a) co-SD AmB:DPPC/DPPG (low P); (b) co-SD AmB:DPPC/DPPG (med P); and (c) co-SD AmB:DPPC/DPPG (high P) (scale bar: 3mm).
Figure 7.
Figure 7.
ATR-FTIR Spectra for: (a) raw AmB; (b) SD AmB (med P); (c) SD AmB (high P); (d) co-SD AmB:DPPC/DPPG (low P); (e) co-SD AmB:DPPC/DPPG (med P); and (f) co-SD AmB:DPPC/DPPG (high P).
Figure 8.
Figure 8.
In Vitro Aerosol Dispersion Performance Using the NGI® Under an Airflow Rate (Q) of 60 L/min with the HandiHaler® Human DPI Device for SD AmB Powders and Co-SD AmB:DPPC/DPPG (n=3, mean ±SD).
Figure 9.
Figure 9.
In vitro Drug Dose-Response Curves for: (a) A549 Cell Dose-Response to Raw AmB, SD AmB, and Co-SD AmB:DPPC/DPPG Powders and (b) H358 Cell Dose-Response to Raw AmB, SD AmB, and Co-SD AmB:DPPC/DPPG Powders; (n=6, mean ± SD).
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