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Toward the Establishment of Standardized In Vitro Tests for Lipid-Based Formulations, Part 6: Effects of Varying Pancreatin and Calcium Levels

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Abstract

The impact of pancreatin and calcium addition on a wide array of lipid-based formulations (LBFs) during in vitro lipolysis, with regard to digestion rates and distribution of the model drug danazol, was investigated. Pancreatin primarily affected the extent of digestion, leaving drug distribution somewhat unaffected. Calcium only affected the extent of digestion slightly but had a major influence on drug distribution, with more drug precipitating at higher calcium levels. This is likely to be caused by a combination of removal of lipolysis products from solution by the formation of calcium soaps and calcium precipitating with bile acids, events known to reduce the solubilizing capacity of LBFs dispersed in biorelevant media. Further, during the digestion of hydrophilic LBFs, like IIIA-LC, the un-ionized–ionized ratio of free fatty acids (FFA) remained unchanged at physiological calcium levels. This makes the titration curves at pH 6.5 representable for digestion. However, caution should be taken when interpreting lipolysis curves of lipophilic LBFs, like I-LC, at pH 6.5, at physiological levels of calcium (1.4 mM); un-ionized–ionized ratio of FFA might change during digestion, rendering the lipolysis curve at pH 6.5 non-representable for the total digestion. The ratio of un-ionized–ionized FFAs can be maintained during digestion by applying non-physiological levels of calcium, resulting in a modified drug distribution with increased drug precipitation. However, as the main objective of the in vitro digestion model is to evaluate drug distribution, which is believed to have an impact on bioavailability in vivo, a physiological level (1.4 mM) of calcium is preferred.

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Acknowledgments

This study results from a joint collaboration between members of the LFCS Consortium, which received funding from Capsugel, Gattefosse, Sanofi, Merck Serono, NiCox, Roche, Bristol-Myers Squibb, and Actelion.

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Authors and Affiliations

  1. Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark

    Philip Sassene, Karen Kleberg & Anette Müllertz

  2. Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Parkville, Victoria, 3052, Australia

    Hywel D. Williams & Christopher J. H. Porter

  3. UMR7282 Enzymologie Interfaciale et de Physiologie de la Lipolyse, Aix Marseille Université, CNRS, 13402, Marseille cedex 20, France

    Jean-Claude Bakala-N’Goma & Frédéric Carrière

  4. Sanofi-Aventis R&D, 371 rue Pr. J. Blayac, 34184, Montpellier, France

    Marilyn Calderone & Ross Blundell

  5. Gattefossé SAS, 36 chemin de Genas, 69804, Saint-Priest, France

    Vincent Jannin & Delphine Marchaud

  6. Capsugel R&D, Strasbourg, France

    Annabel Igonin, Jan Vertommen & Hassan Benameur

  7. Merck KGaA, Darmstadt, Germany

    Anette Partheil & Mario Maio

  8. Capsugel Product Development Center, Boston, Massachusetts, USA

    Eduardo Jule

  9. Pharmaceutical Biology, Monash Institute of Pharmaceutical Sciences, Parkville, Victoria, 3052, Australia

    Colin W. Pouton

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  1. Philip Sassene
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Correspondence to Anette Müllertz.

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Sassene, P., Kleberg, K., Williams, H.D. et al. Toward the Establishment of Standardized In Vitro Tests for Lipid-Based Formulations, Part 6: Effects of Varying Pancreatin and Calcium Levels. AAPS J 16, 1344–1357 (2014). https://doi.org/10.1208/s12248-014-9672-x

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