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In vitro model development of drug-microbes interaction in colonic mucus environment

Terrmin

HT23

Beskrivning

Aim
To screen model bacteria and lipid vesicles mimicking drug bioaccumulation phenomenon which present in colonic mucus environment

Background
Translation of in vitro research into in vivo clinical relevance is currently still an issue, with time and effort spent in drug discovery and pre-clinical step (Matthews et al. 2016). Development of an efficient drug testing platform is necessary in accordance with ethical 3R principles (reduce, replace, refine animal experiments) and sustainable development goals 2030 of the UN. In comparison to the small intestine, the colon has biological properties which pose significant hurdles for the drug delivery system. These include in particular the extensive colonic microbiota and the thicker mucus barrier (Naeem et al. 2020). In this study, focus is set on the characteristics of microbes' role in the mucus barrier which might influence drug diffusion and off-target bioaccumulation. By establishing a relevant in vitro system of microbes or lipid vesicles model bioaccumulation in biosimilar mucus models for drug testing, a proper estimate of in vivo performance of the delivery system can be made based on lab bench methods only.

Methods
- Constructing bacterial membranes model with lipid vesicles/liposomes
- Using DLS, plate reader and/or HPLC to reveal the drug-membrane / drug-microbes interaction and quantify drug concentration
- Measure and calculate the fraction bound (Fb) & the fraction unbound (Fu) of a series of model compounds
- Permeation studies with Transwell (API solution vs API-liposome solution) to see if the binding to liposome will affect the drug permeability, using Biosimilar Porcine Artificial Colonic Mucus (PACM) prototypes established from previous project
- Testing of microbes viability in PACM
- Testing of microbes influence toward viscosity & storage modulus of PACM with rheometer
- Microbes and drug diffusion determination based on confocal microscopy of Fluorescence Recovery After Photobleaching (FRAP) / Multiple Particle Tracking (MPT)

My task
- Write project plan and report
- Perform the experiments involving liposome and PACM synthesize, and evaluate with rheometer, DLS, FRAP/MPT, plate reader, HPLC and permeation studies in transwell system

Expected results:
Permeability profile of a series of structurally diverse compounds
Diffusion-bioaccumulation profile of a series of structurally diverse drug compound
Established liposome as model for in vitro colonic drug bioaccumulation
Fb and Fu of the model compounds
Viscosity and storage modulus profile of PACM-microbes

Huvudområde

Läkemedelsutveckling

Ämne

Läkemedelsformulering och Molekylär galenisk farmaci

Typ

Laborativ studie

Företag

Uppsala universitet

Ort/Plats

Uppsala

Handledarens namn

Christel Bergström

Handledarens e-post

christel.bergstrom@farmaci.uu.se

Institution

Institutionen för farmaci

Program

Masterprogram i läkemedelsutveckling

Kurs

Omfattning/hp

45hp