Extending the possibilities of biopartitioning chromatography for improved prediction of in vivo drug absorption Mike De Vrieze Promotor: Prof. Dr. F. Lynen Co-promotor: Prof. Dr. P. Sandra 2015 Thesis submitted to obtain the degree of Doctor in Sciences, Chemistry Extending the possibilities of biopartitioning chromatography for improved prediction of in vivo drug absorption Mike De Vrieze Promotor: Prof. Dr. F. Lynen Co-promotor: Prof. Dr. P. Sandra 2015 Thesis submitted to obtain the degree of Doctor in Sciences, Chemistry Table of contents Table of contents Chapter I. General introduction and aims of the work ..................................... 1 I.1 References ................................................................................................................. 5 Chapter II. Description of the structure, function and delivery across the Blood-Brain Barrier ............................................................................................... 7 II.1 Structure and function of the blood-brain barrier ............................................... 7 II.1.1 Transport across the blood-brain barrier ....................................................................... 12 II.1.2 Functions of the blood-brain barrier .............................................................................. 15 II.1.3 Controlling the permeability of the blood-brain barrier ................................................ 15 II.2 Principles of drug delivery across the blood-brain barrier ................................ 17 II.2.1 Non-mediated delivery .................................................................................................. 17 II.2.2 Drug delivery across the blood-brain barrier by liposome systems .............................. 19 II.2.3 Bicelles as an alternative delivery system ..................................................................... 20 II.2.4 Cell-penetrating peptides and molecular Trojan horses ................................................ 22 II.2.5 Drug delivery through nanocarriers ............................................................................... 22 II.2.6 Tight junction opening using focused ultrasound .......................................................... 23 II.2.7 Intranasal delivery as a drug delivery system ................................................................ 24 II.2.8 Other delivery systems .................................................................................................. 25 II.3 Measurement and prediction of drug transport across the blood-brain barrier .................................................................................................................................. 26 II.3.1 In vivo methods for assessing blood-brain barrier permeability ................................... 27 II.3.1.1 Determination of the logarithm of the brain versus blood ratio (log BB) ............. 28 II.3.1.2 Measurement of the permeability-surface area product (PSP) .............................. 28 II.3.1.3 Microdialysis for the measurement of brain penetration ....................................... 29 II.3.1.4 The measurement of cerebrospinal fluid (CSF) concentrations ............................ 30 II.3.1.5 In situ perfusion to study drug absorption ............................................................. 31 II.3.1.6 Other in vivo methods ........................................................................................... 32 II.3.2 In vitro systems for the prediction of drug transport ..................................................... 32 i Table of contents II.3.2.1 High-performance liquid chromatography ............................................................ 32 II.3.2.1.1 Micellar liquid chromatography for the prediction of drug transport .............. 33 II.3.2.1.2 The use of microemulsion liquid chromatography to assess membrane permeation .......................................................................................................................... 36 II.3.2.1.3 Immobilized Artificial Membrane Liquid Chromatography for the prediction of drug transport ....................................................................................................................... 37 II.3.2.1.4 Other HPLC-based techniques ......................................................................... 43 II.3.2.2 The assessment of membrane permeation by capillary electrophoresis ................ 45 II.3.2.3 Parallel artificial membrane permeation assay to predict drug transport .............. 48 II.3.2.4 Bio-mimetic artificial membrane permeation to predict drug transport ................ 48 II.3.2.5 The use of cell lines for the prediction of intestinal absorption ............................ 49 II.3.2.6 The use of Madin-Darby canine kidney cells for the prediction of drug transport 50 II.3.2.7 Other methods ....................................................................................................... 50 II.3.3 The prediction of drug transport across the blood-brain barrier with in silico systems 52 II.3.3.1 Examples of in silico predictions of log BB values ............................................... 52 II.3.3.2 Classification systems for the prediction of drug transport ................................... 55 II.4 Regression models for the prediction of blood-brain barrier transport ........... 55 II.4.1 Computational models to establish relationships between descriptors and values of interest. ....................................................................................................................................... 55 II.4.2 The use of QSRR, QSAR and QRAR ........................................................................... 57 II.4.3 Validation of the computational models ........................................................................ 58 II.4.4 Model selection ............................................................................................................. 59 II.5 References ............................................................................................................... 60 Chapter III. Predicting drug penetration across the blood–brain barrier: comparison of micellar liquid chromatography and immobilized artificial membrane liquid chromatography ...................................................................... 77 III.1 Summary ................................................................................................................. 77 III.2 Introduction ............................................................................................................ 78 III.3 Materials and methods ........................................................................................... 79 ii Table of contents III.3.1 Chemicals ...................................................................................................................... 79 III.3.2 Apparatus ....................................................................................................................... 79 III.3.3 Mobile phase and sample preparation ........................................................................... 80 III.3.4 Data sources, software, and processing ......................................................................... 81 III.4 Rationale ................................................................................................................. 81 III.5 Results and discussion ............................................................................................ 84 III.5.1 Prediction of log BB ...................................................................................................... 85 III.6 Conclusion ............................................................................................................... 91 III.7 References ............................................................................................................... 92 Chapter IV. Development of the first Sphingomyelin biomimetic stationary phase for Immobilized Artificial Membrane Liquid Chromatography ............... 95 IV.1 Summary ................................................................................................................. 95 IV.2 Introduction ............................................................................................................ 95 IV.3 Materials and methods ........................................................................................... 97 IV.3.1 Chemicals ...................................................................................................................... 97 IV.3.2 Apparatus ....................................................................................................................... 97 IV.3.3 Experimental conditions ................................................................................................ 98 IV.3.4 Data Sources, software, and processing ........................................................................ 99 IV.4 Results and discussion ............................................................................................ 99 IV.5 Conclusion ............................................................................................................. 102 IV.6 References ............................................................................................................. 104 Chapter V. Evaluation of sphingomyelin, cholester and phosphatidylcholine based immobilized artificial membrane liquid chromatography