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Target Screening for Pharmaceuticals – cell-free GPCR drug discovery high-throughput screening

Pharmaceutical market The top 20 players in pharmaceutical industry had revenues of US$ 497,519 million in 2006. In 2005 a global market growth of 7 % was achieved. The pharmaceutical market is still expanding.

GPCRs (G protein-coupled receptor) take up a significant share of drug development pipeline across the pharmaceutical industry.

GPCRs: 40 % of pharmaceutical targets GPCR is the largest known category of molecular targets with verified therapeutic value. Up to 40% of approved drugs are targeted to GPCRs. 25% of the top 200 best-selling drug targets are GPCRs.

GPCR ligand screening Screening for GPCR-targeted drugs thus is a hot topic in pharmaceutical research. The very high test turnover allows screening of 300.000 substances per day (Bayer Health Care).

Chemical compound library screening Our assay can be implemented in a cell-free chemical compound high-throughput target screening.

INVENTION / Technology offer

GPCR ligand screening assay

Monitors the very early activation stage of Frizzled-Receptors of the GPCR group.

Patent Status:

The US Patent is granted, an European and a US divisional patent application are currently pending.

Monitors very early activation stage We have established an assay which monitors the very early stages of Wnt/Frizzled activation - namely the activation of trimeric G proteins in response to Wnt ligand-induced activation of the Frizzled receptors.

As 19 Wnt ligands and 10 Frizzled receptors exist in humans, our assay can be targeted to any of the particular Wnt/Frizzled pair and thus be extremely specific for any particular case of pathological or physiological activation of the pathway.

Furthermore, our assay can identify small molecules which do not need membrane permeability, thus markedly expanding the molecule repertoire from which to screen for agonists/antagonists serving as future drugs.

Your benefit • very simple & robust • upgrade for high throughput screening • chemical compound library screening • monitoring very early activation GPCR

stage • cell-free : no side-effects

GPCR Market • 25% of the top 200 best-selling drug

targets • 40% of approved drugs • Highest test turnover of 300.000

substances per day

GPCRs as talented all-rounders Involved in

• Alzheimer • Cancer • Schizophrenia • Virus, also tumor virus actions • Stem cells • All fundamental biological processes

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State of the Art The available platforms to screen for agonists/antagonists of the Wnt/Frizzled pathways mainly use cell culture-based assays, where signal transduction is monitored by activation of reporter constructs placed under the beta-catenin-dependent transcriptional regulation.

Disadvantages of the State of the Art The disadvantage of this assay is that it is prone to identify agents acting at the downstream levels of signaling, which are conserved among multiple Wnt/Frizzled sub-pathways - and thus lacking selectivity amongst this multitude of pathways.

INVENTION:

Frizzled GPCR ligand screening assay

The assay is very simple and robust. It comprises recombinant human Wnt ligands and Frizzled receptors, bovine G proteins, and guanine nucleotide analogs modified for time-resolved fluorescence (usage of other ways of labeling guanine nucleotide analogs such as for radioactive or fluorescent measurements is also possible).

In a high-throughput format, chemical libraries are added to the assay to screen for antagonists of the Wnt/Frizzled-induced activation of G proteins, or for agonists of Frizzled receptors (which would activate the G proteins in the absence of Wnt ligands but presence of Frizzled receptors).

Wnt/Frizzled signaling pathways The Wnt/Frizzled signaling pathways are highly conserved in animal evolution and control multiple steps during organism development. In the healthy adult, these pathways are mostly silent.

The physiological and pathological exceptions of this silence are of the paramount medical importance.

Candidates for the treatment of Alzheimer, Cancer, Schizophrenia Thus, both agonists (activators) and antagonists (inhibitors) of the Wnt/Frizzled pathway are highly demanded as therapeutic agents, the first being candidates for the regenerative medicine and anti-Alzheimer’s drugs, as well as agents for in vitro stem cell proliferation; the antagonists being candidates for the anti-cancer drugs, as well as anti-schizophrenia agents, and even “pills of youth”.

Cancer: Inappropriate activation of the Wnt/Frizzled pathways leads to cancerogenesis in all tissues; the most prominent examples are the colon cancer (ca. 80% of cases due to overactivation of the pathway) and breast cancer (ca. 50% of cases due to overactivation of the pathway).

Schizophrenia: Another pathological consequence of overactivation of the Wnt/Frizzled pathway is schizophrenia. Aging: Furthermore, whole-body activation of this pathway has been linked to aging. Stem cells/organ regeneration: In contrast, activation of this pathway is necessary for the proliferation of various types of stem cells (hematopoetic, neuronal, embryonic, etc) and has been shown necessary for organ regeneration. Alzheimer: Inhibition of this pathway has also been implicated in the beta-amyloid toxicity in Alzheimer’s disease. Selected rerefence Koval A, Purvanov V, Egger-Adam D, Katanaev VL. (2011) Yellow Submarine of the Wnt/Frizzled Signaling: Submerging from the G Protein Harbor to the Targets. Biochemical Pharmacology 82: 1311-1319.

Contact: Andrea Nestl, PhD nestl@tlb.de Technology Licence Office (TLB) GmbH of the Universities in Baden-Württemberg, Ettlinger Straße 25, 76137 Karlsruhe, Germany Tel +49 721 79004-0, Fax +49 721 79004-79 www.tlb.de, http://twitter.com/TLBGmbh

http://de-de.facebook.com/TLBGmbH

• State of the Art: cell culture-based

assays,disadvantage: lacking selectivity

• Invention: cell-free & specific

Technology Transfer

The Technologie-Lizenz-Büro GmbH is charged with the commercialization of this techology and now offers companies the opportunity to enter into a cooperation and/or licensing agreement