Completed R&D Projects

BMBF Program "Innovation in drug development“:

In vitro model for hepatic and extra-hepatic drug metabolism  and toxicity  (HepaTox)

A major hurdle in preclinical drug testing is the existing lack in suitable in vitro models that allow the prediction of potentially toxic effects in humans. The project is aimed at providing a novel in vitro model to analyze human-specific hepatotoxic effects of drugs and of hepatic-generated intermediates in human liver cells.

Two models will be tested: a Petri-Dish 2D culture model and a 3D-multicompartment bioreactor tech-nology that allows the maintenance of human liver cells in a perfused, tightly controlled environment with high mass exchange and decentralized oxygenation, thereby creating physiological condi-tions for the cells similar to the situation within the liver in vivo. Readout parameters for assessing potential hepatotoxic effects in the models will be established based on a multilevel concept that includes parameters for basal toxicity, for general and specific functionality, morphological criteria and complex metabolic pathways. The response of the cultures to drug exposition in 3D bioreactor cultures will be modelled based on systems biotechnological analysis.

Data will be compared for validation to in vitro data from 2D culture models and to in vivo data available within the consortium and/or in public databases. In case of successful model development, commercial exploitation of the new methodology will be brought forwards by the industry partners.

Publication

EU-FP6: Cardioworkbench - Drug design for cardiovascular diseases: Integration of in silico and in vitro analyses

The aim of the FP6 project called CARDIOWORKBENCH was to identify novel factors contributing to cardiovascular diseases such as ischemic heart failure and to set up new systems and strategies to select and test pharmacologically active molecules with therapeutic potential. Within the three year funding period, partners of the project successfully identified two putative targets related to ischemic heart failure. Pharmacelsus together with six other partners managed to synthesize and characterize a full range of potential drug candidates. The joint efforts culminated into a new approach to inhibit Rac1 as a target to treat heart failure. The resulting intellectual property was entered into a European patent application entitled “New Rac1 inhibitors as potential pharmacological agents for heart failure treatment” (No: EP09157855).

Cardioworkbench

EU FP6: InVitroHeart: Reducing animal experimentation in drug testing by human cardiomyocyte in vitro models derived from embryonic stem cells

The objective of the project was to establish stable cell lines that reliably reflect human cardiomyocyte properties by the development of models derived from human embryonic stem cells. The aims were (1) to deliver reliable in vitro models that could be used by the pharmaceutical industry to replace experimental animals in investigations on pharmacological toxicity and safety of compounds in the drug discovery and development processes, and (2) the testing of toxic effects of chemicals according to the new system of the Community on the Registration, Evaluation and Authorisation of Chemicals (REACH).

InVitroHeart

EU-FP6:  Vitrocellomics:     Reducing animal experimentation in preclinical predictive drug testing by human hepatic in vitro models derived from embryonic stem cells

The aim of the FP6 project VITROCELLOMICS was to deliver reliable in vitro models that could be used by the pharmaceutical industry for investigations on human hepatic drug metabolism and toxicology properties of compounds in the drug discovery and development process. The isoforms of the cytochrome P450 (CYP) family are the key enzymes of drug metabolism and induction of a CYP isoenzyme is one of the major reasons for drug-drug interactions. During the project, Pharmacelsus succeeded in developing a CYP450 induction assay for fresh and cryopreserved primary cultured human hepatocytes as well as for the commercially available hepatoma cell line HepaRG® as alternative to the rarely available human primary hepatocytes.

Vitrocellomics

ECVAM:     Multi-study validation trial for cytochrome P450 induction providing a standard for human hepatic metabolism and toxicity using the human HepaRG® cell line and cryopreserved human hepatocytes

Primary hepatocytes derived from human liver tissue are considered as the gold standard for drug-drug interaction studies. However the availability of freshly isolated human hepatocytes is limited, their isolation is an elaborate procedure and they have a limited survival time (Bjornsson et al., 2003). The commercially available hepatoma cell line HepaRG®, derived from a human hepatocellular carcinoma, was shown to maintain hepatic functions and expression of liver-specific genes, such as P450s, Phase II enzymes, hepatic drug transporters and nuclear receptors (Aninat et al., 2006; Le Vee et al., 2006; Kanebratt & Anderson, 2008). During the EU FP6 project Vitrocellomics, the consortium was able to enter in a pre-validation study under the auspices of the European Centre for Validation of Alternative Methods (ECVAM) using the HepaRG® cell line as an alternative tool to primary human hepatocytes. Pharmacelsus was dedicated to be the lead lab of the prevalidation study and intensively cooperated with ECVAM. In 2010, the prevalidation became a part of a formal multi-study validation trial in cooperation with ECVAM and laboratories in Italy, Belgium, France and Sweden. The HepaRG® induction method itself was validated regarding inter- and intra-assay variability as well as inter- and intra-batch variability.