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Transdermal Delivery of Cytochrome C—A 12.4 kDa Protein—Across Intact Skin by Constant–Current Iontophoresis

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Abstract

Purpose

To demonstrate the transdermal iontophoretic delivery of a small (12.4 kDa) protein across intact skin.

Materials and Methods

The iontophoretic transport of Cytochrome c (Cyt c) across porcine ear skin in vitro was investigated and quantified by HPLC. The effect of protein concentration (0.35 and 0.7 mM), current density (0.15, 0.3 or 0.5 mA.cm−2 applied for 8 h) and competing ions was evaluated. Co-iontophoresis of acetaminophen was employed to quantify the respective contributions of electromigration (EM) and electroosmosis (EO).

Results

The data confirmed the transdermal iontophoretic delivery of intact Cyt c. Electromigration was the principal transport mechanism, accounting for ∼90% of delivery; correlation between EM flux and electrophoretic mobility was consistent with earlier results using small molecules. Modest EO inhibition was observed at 0.5 mA.cm−2. Cumulative permeation at 0.3 and 0.5 mA.cm−2 was significantly greater than that at 0.15 mA.cm−2; fluxes using 0.35 and 0.7 mM Cyt c in the absence of competing ions (J tot  = 182.8 ± 56.8 and 265.2 ± 149.1 μg.cm−2.h−1, respectively) were statistically equivalent. Formulation in PBS (pH 8.2) confirmed the impact of competing charge carriers; inclusion of ∼170 mM Na+ resulted in a 3.9-fold decrease in total flux.

Conclusions

Significant amounts (∼0.9 mg.cm−2 over 8 h) of Cyt c were delivered non-invasively across intact skin by transdermal electrotransport.

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Abbreviations

ACE:

acetaminophen

Cyt c:

cytochrome c

CZE:

capillary zone electrophoresis

EM:

electromigration

EO:

electroosmosis

JEM, Cyt c :

flux due to electromigration of Cyt c

JEO, Cyt c :

flux due to electroosmosis of Cyt c

Jtot, Cyt c :

total steady-state flux of Cyt c

Vw :

convective solvent flow

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Acknowledgements

We would like to thank Prof. Leonardo Scapozza and Dr. Shaheen Ahmed for help with the molecular graphics representations of Cytochrome c. J. Cázares Delgadillo acknowledges support from CONACYT (Mexico).

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Author notes

  1. A. Naik

    Present address: Triskel Integrated Services SA, 4 rue des Terreaux-du-Temple, 1201, Geneva, Switzerland

Authors and Affiliations

  1. School of Pharmaceutical Sciences, University of Geneva & University of Lausanne, 30 Quai Ernest Ansermet, 1211, Geneva, Switzerland

    J. Cázares-Delgadillo, A. Naik & Y. N. Kalia

  2. “Pharmapeptides”, Centre Interuniversitaire de Recherche et d’Enseignement, 74160, Archamps, France

    J. Cázares-Delgadillo, A. Naik & Y. N. Kalia

  3. División de Estudios de Posgrado (Tecnología Farmacéutica), Facultad de Estudios Superiores Cuautitlán, Universidad Nacional Autónoma de México, Av. 1° de Mayo S/N Cuautitlán Izcalli, 54704, Estado de México, Mexico

    A. Ganem-Rondero & D. Quintanar-Guerrero

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  1. J. Cázares-Delgadillo
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Correspondence to Y. N. Kalia.

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Cázares-Delgadillo, J., Naik, A., Ganem-Rondero, A. et al. Transdermal Delivery of Cytochrome C—A 12.4 kDa Protein—Across Intact Skin by Constant–Current Iontophoresis. Pharm Res 24, 1360–1368 (2007). https://doi.org/10.1007/s11095-007-9294-4

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  • DOI: https://doi.org/10.1007/s11095-007-9294-4

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