Over the last decade, surveys in European and US hospitals have shown that more than 60% of patients in pediatric wards receive one or more prescriptions unlicensed or off-label for children. Despite important progress, several drugs are still unavailable in oral formulations suitable for infants and children. Our preliminary experiments show that the casein micelles from bovine milk can be modified to (1) bind low molecular weight (MW) hydrophobic molecules, and (2) dried to a milk-based powder and re-suspended in an aqueous phase to permanently disperse hydrophobic molecules. The objective of this proposal is to use modified casein micelles from bovine milk as carrier systems for the delivery of the low MW hydrophobic ritonavir (MW = 721 KDa, Log P >3.9) into the small intestine. The central hypothesis is that the casein micelle can be modified to facilitate the intestinal absorption of hydrophobic molecules. This hypothesis is formulated based on exploratory experiments showing that (1) modified casein micelles strongly bind the hydrophobic molecule triclosan, (2) little triclosan is released in the 3-5 pH range and, (3) release is favored at pH e 7. Both in vitro and in vivo experiments are needed to determine the key physicochemical mechanisms that promote load, the release kinetics of loaded micelles, and the effect of gastric enzymes on casein-drug systems. In order to achieve this objective we have assemble a multidisciplinary team that has the required breadth and scope of experience. We plan to test our central hypothesis by pursuing the following four specific aims:
Specific aim 1 : Optimize the physicochemical conditions that maximize casein micelle association to specific ritonavir polymorphs.;
Specific aim 2 : Determine the release kinetics of ritonavir in vitro model systems simulating stomach and small intestine;
Specific aim 3 : Determine bioavailability following casein-ritonavir administration in a young pig model, and compare it to the commercially available formulation (i.e., Norvir). The rationale for the proposed research is that by capitalizing on the casein micelle's carrying capabilities, we will be able to develop milk-based ethanol-free powder systems to disperse hydrophobic drugs and improve bioavailability of oral formulations targeting infant and child populations.
The Best Pharmaceuticals for Children Act  and the European Pediatric Formulation Initiative  were created to tackle the need for pharmacokinetic and pharmacodynamic data for drugs used in infant and child populations and to push for drug formulations specifically designed for pediatric use. Despite important progress, several drugs are still unavailable in oral formulations suitable for infants and children. For the case of orally administered hydrophobic drugs, there is a need for alternative delivery systems to (1) disperse the drugs in aqueous phases, (2) bypass the gastric environment, delivering the drug in the early portions of the small intestine, and (3) mask off-flavors. This research intends to explore the use of a naturally occurring nano-delivery system, the casein micelle from bovine milk, as a delivery system for hydrophobic drugs that are orally administered to infants and children. The long term product in a successful outcome of this research project will be a milk-based drug delivery system that can be used to formulate hydrophobic drugs as readily dispersible milk-powder for the oral administration as fluids to children and infants in a regular basis. This research also opens new grounds for the development of drug carrier systems using protein quaternary structures.
|Corzo-Martínez, M; Mohan, M; Dunlap, J et al. (2015) Effect of ultra-high pressure homogenization on the interaction between bovine casein micelles and ritonavir. Pharm Res 32:1055-71|
|Ye, Ran; Harte, Federico (2014) High pressure homogenization to improve the stability of casein - hydroxypropyl cellulose aqueous systems. Food Hydrocoll 35:670-677|
|Mohan, M S; Jurat-Fuentes, J L; Harte, F (2013) Binding of vitamin A by casein micelles in commercial skim milk. J Dairy Sci 96:790-8|
|Ye, Ran; Harte, Federico (2013) Casein maps: effect of ethanol, pH, temperature, and CaCl2 on the particle size of reconstituted casein micelles. J Dairy Sci 96:799-805|
|Trejo, R; Dokland, T; Jurat-Fuentes, J et al. (2011) Cryo-transmission electron tomography of native casein micelles from bovine milk. J Dairy Sci 94:5770-5|