Epithelia transmembrane transport of orally administered ultrafine drug particles evidenced by environment sensitive fluorophores in cellular and animal studies.

Little is known about the in vivo fate of drug particles taken orally, in particular, the drug release kinetics and interaction with the gastrointestinal (GI) membrane. Lacking is analytical means that can reliably identify the integrity of drug particles under the complexity of biological environment. Herein, we explored fluorescent probes whose signals become quenched upon being released from drug carriers. Taking advantage of so-called the aggregation caused quenching (ACQ), particles may be identified by the integrated fluorophores, which are "turned off" when the particles become destructed and dyes are released. In the current study, ultrafine amorphous particles (UAPs) of cyclosporin A (CsA) were prepared with synthesized ACQ dyes physically entrapped. The fluorescence intensity of suspension of these UAPs was found correlated well with the dissolution of the particles. When given to rats orally, it was found that some of the administered UAPs could survive the animal's GI tracts for as long as 18h. Whole-body fluorescence imaging detected fluorescent signals in the liver and lungs. Particularly noticed in sections of jejunum and ileum, the detection suggested the possibility of direct absorption of UAPs through epithelial membranes. Moreover, 250nm particles were absorbed faster via transepithelia than larger ones (550nm), while the latter were preferably taken up by M cells in the follicle-associated epithelium (FAE) region of Peyer's patches. In vitro permeation studies with Caco-2 cells confirmed the transmembrane transport of the dye-integrated UAPs. Our study supports the idea of using ACQ fluorophores for imaging and characterizing the fate of intact particles in a biological environment.