Involvement of Endocytosis in the Transdermal Penetration Mechanism of Ketoprofen Nanoparticles.

Noriaki Nagai,Yosuke Nakazawa,Miyu Ishii,Naohito Kawasaki,Fumihiko Ogata,Hiroko Otake,Yuya Fukuoka

doi:10.3390/ijms19072138

Noriaki Nagai, Yosuke Nakazawa + Show 5 more

Open Access

https://doi.org/10.3390/ijms19072138

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Abstract

We previously designed a novel transdermal formulation containing ketoprofen solid nanoparticles (KET-NPs formulation), and showed that the skin penetration from the KET-NPs formulation was higher than that of a transdermal formulation containing ketoprofen microparticles (KET-MPs formulation). However, the precise mechanism for the skin penetration from the KET-NPs formulation was not clear. In this study we investigated whether energy-dependent endocytosis relates to the transdermal delivery from a 1.5% KET-NPs formulation. Transdermal formulations were prepared by a bead mill method using additives including methylcellulose and carbopol 934. The mean particle size of the ketoprofen nanoparticles was 98.3 nm. Four inhibitors of endocytosis dissolved in 0.5% DMSO (54 μM nystatin, a caveolae-mediated endocytosis inhibitor; 40 μM dynasore, a clathrin-mediated endocytosis inhibitor; 2 μM rottlerin, a macropinocytosis inhibitor; 10 μM cytochalasin D, a phagocytosis inhibitor) were used in this study. In the transdermal penetration study using a Franz diffusion cell, skin penetration through rat skin treated with cytochalasin D was similar to the control (DMSO) group. In contrast to the results for cytochalasin D, skin penetration from the KET-NPs formulation was significantly decreased by treatment with nystatin, dynasore or rottlerin with penetrated ketoprofen concentration-time curves (AUC) values 65%, 69% and 73% of control, respectively. Furthermore, multi-treatment with all three inhibitors (nystatin, dynasore and rottlerin) strongly suppressed the skin penetration from the KET-NPs formulation with an AUC value 13.4% that of the control. In conclusion, we found that caveolae-mediated endocytosis, clathrin-mediated endocytosis and macropinocytosis are all related to the skin penetration from the KET-NPs formulation. These findings provide significant information for the design of nanomedicines in transdermal formulations.

Highlights

Ketoprofen is a non-steroidal anti-inflammatory drug (NSAID) that is scarcely soluble in water with a log partition coefficient are 0.185 mg/mL, 3.11, respectively
We investigated the effect of endocytosis on the skin penetration of the KET-NPs formulation, and found that three endocytosis pathways (CavME, clathrin-mediated endocytosis (CME) and MP) are related to the skin penetration of ketoprofen nanoparticles
It has previously been reported that nanoparticles with diameters in the range of 60–100 nm are optimal for the cellular uptake process [17,18,19]

Summary

Introduction

Ketoprofen is a non-steroidal anti-inflammatory drug (NSAID) that is scarcely soluble in water with a log partition coefficient are 0.185 mg/mL, 3.11, respectively. One promising method to reduce the adverse effects is to deliver the drug through the skin. Due to the excellent barrier function of the skin, the need to use safe and effective enhancers to improving the transdermal absorption of drugs is well recognized [5,6,7,8]. We designed transdermal formulations containing ketoprofen nanoparticles (KET-NPs formulation, particle size approximately 80–200 nm), and showed a high accumulation of ketoprofen released from the KET-NPs formulation in skin tissues. The therapeutic effect on inflammation of the KET-NPs formulation is significantly greater than that of commercially available ketoprofen ointments (dissolution type) [20]. The design of a KET-NPs formulation may represent a novel transdermal delivery system for the management of inflammation. It is important to understand the mechanism of transdermal penetration from the KET-NPs formulation

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