The NCCR TransCure screening, profiling and analytical facility (SPAF), which is located at the IBMM University of Bern, provides the workforce, expertise and equipment required to develop and validate screening assays for diverse transporter targets. We assist PIs from the individual projects to upscale their assays to use them to evaluate small molecules in a typical medium-throughput format (<6’000 compounds) for primary target activity, and if required for selectivity studies. Compounds are provided by the chemistry groups either as targeted sets selected by virtual screening, from CROs, or as series of focused synthetic compounds generated in house or in collaboration with industry partners. More importantly, the SPAF offers a unique service at the University of Bern to investigate bioavailability, tissue distribution. pharmacokinetics and pharmacodynamics of probes and drug leads in vivo in different preclinical models by using high-end LC-MS/MS analytics. Current project with links to the screening facility are LAT1, DMT1, CaSR, endocannabinoid transport (collaboration Synendos Therapeutics), and TRPM4. In-house facilities for screening include basic 96-well robotics for compound handling if required; a FLIPR automated 96-well parallel fluorescence plate reader, plate washers, and a TopCount NXT 96-well scintillation counter. We have collaborated with Novartis for high-throughput screening assays (HTS). Our chemical tools and probes are fundamental to study biological phenomena. Screening and profiling have led to the optimization of TRPV6 calcium channel and TRPM4 sodium channel inhibitors by using a 3D ligand-based virtual screening method. Moreover, the SPAF was instrumental for the development of selective endocannabinoid reuptake inhibitors and the new generation LAT1 inhibitors. To address analytical problems for the in vivo proof of concept, we employ LC-MS/MS (Sciex 4000 and 5500) to quantify tool compounds in vivo. A future focus of the SPA is to engage in translational research. The SPAF has made fundamental contributions to better pharmacology and will continue its activity beyond NCCR TransCure.

Relevant literature

Graff J, Müller J, Sadurní A, Rubin M, Canivete Cuissa IA, Keller C, Hartmann M, Singer S, Gertsch J, Altmann KH. The Evaluation of l-Tryptophan Derivatives as Inhibitors of the l-Type Amino Acid Transporter LAT1 (SLC7A5). ChemMedChem. 2022 Jul 27:e202200308. doi: 10.1002/cmdc.202200308. Online ahead of print.

Yan R, Li Y, Müller J, Zhang Y, Singer S, Xia L, Zhong X, Gertsch J, Altmann KH, Zhou Q. Mechanism of substrate transport and inhibition of the human LAT1-4F2hc amino acid transporter. Cell Discov. 2021 Mar 23;7(1):16.

Mäder P, Bartholomäus R, Nicolussi S, Baumann A, Weis M, Chicca A, Rau M, Simão AC, Gertsch J, Altmann KH. Synthesis and Biological Evaluation of Endocannabinoid Uptake Inhibitors Derived from WOBE437. ChemMedChem. 2021 Jan 8;16(1):145-154.

Häfliger P, Graff J, Rubin M, Stooss A, Dettmer MS, Altmann KH, Gertsch J, Charles RP. The LAT1 inhibitor JPH203 reduces growth of thyroid carcinoma in a fully immunocompetent mouse model. J Exp Clin Cancer Res. 2018 Sep 21;37(1):234.

Ozhathil LC, Delalande C, Bianchi B, Nemeth G, Kappel S, Thomet U, Ross-Kaschitza D, Simonin C, Rubin M, Gertsch J, Lochner M, Peinelt C, Reymond JL, Abriel H. (2018), Identification of potent and selective small molecule inhibitors of the cation channel TRPM4. Br J Pharmacol.175(12):2504-2519.

Chicca A, Nicolussi S, Bartholomäus R, Blunder M, Aparisi Rey A, Petrucci V, Reynoso-Moreno IDC, Viveros-Paredes JM, Dalghi Gens M, Lutz B, Schiöth HB, Soeberdt M, Abels C, Charles RP, Altmann KH, Gertsch J. Chemical probes to potently and selectively inhibit endocannabinoid cellular reuptake. Proc Natl Acad Sci U S A. 2017 Jun 20;114(25):E5006-E5015.

Simonin C, Awale M, Brand M, van Deursen R, Schwartz J, Fine M, Kovacs G, Häfliger P, Gyimesi G, Sithampari A, Charles RP, Hediger MA, Reymond JL. (2015) Optimization of TRPV6 Calcium Channel Inhibitors Using a 3D Ligand-Based Virtual Screening Method. Angew Chem Int Ed Engl., doi: 10.1002/anie.201507320.

Ilgü H, Jeckelmann JM, Gachet MS, Boggavarapu R, Ucurum Z, Gertsch J, Fotiadis D. Variation of the detergent-binding capacity and phospholipid content of membrane proteins when purified in different detergents. Biophys J. 2014 Apr 15;106(8):1660-70.