Methods for determining the efficacy profile of a drug candidate

Abstract

This patent application relates to a method for determining the in vitro efficacy profile of a drug candidate using standardized cell cultures of uniformly distributed differentiated neural cells (NCs) from at least two primate species, wherein the differentiated NC cultures are qualified for high throughput screening based on a dissociation and reseeding step performed on the differentiated NCs. The method includes differentiating human and / or non-human primate neuronal precursor cells (NPCs) to neuronal cells (NCs) followed by dissociating the differentiated NCs from its support and reseeding the differentiated NCs in a high-throughput cell culture format resulting in robust cultures suitable for high-throughput drug screening assays, in particular to screen antisense oligonucleotides.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation of PCT International Application No. PCT / EP2016 / 077435, filed on Nov. 11, 2016, the entire contents of which are incorporated herein in by reference, and which claims priority to European Patent Application No. 16189502.4 filed on Sep. 19, 2016 and European Patent Application No. 15194367.7 filed on Nov. 12, 2015.SEQUENCE LISTING[0002]This application contains a Sequence Listing submitted via EFS-Web and hereby incorporated by reference in its entirety. Said ASCII copy, created on Apr. 20, 2018, is named P33890-US_SeqListing.txt, and is 554,456 bytes in size.FIELD OF THE INVENTION[0003]This patent application relates to a method for determining the in vitro efficacy profile of a drug candidate using standardized cell cultures of uniformly distributed differentiated neural cells (NCs) from at least two primate species, wherein the differentiated NC cultures are qualified for high throughput screening base...

AI Technical Summary

Benefits of technology

This approach enables reproducible and standardized high-throughput screening, improving the accuracy of drug candidate selection by providing uniform and viable neural cell cultures for inter-species comparison, thereby reducing the need for animal testing and enhancing the prediction of in vivo efficacy and toxicity.

Problems solved by technology

While the in vitro testing of drug candidates allows to reduce the number of laboratory animals sacrificed for drug testing, it also posed challenges of outcome translatability from in vitro to in vivo and to the human physiology.

Indeed, the translatability of the efficacy and toxicity profiles of drug candidates from rodent species to humans can be challenging, due to non-close genetic relation between the species.

On the other hand, the use of NHP species in drug discovery remains a controversial matter.

However, there are formidable hurdles for gaining such primate inter-species transferable efficacy and toxicity data in vitro from stem cell-derived differentiated NCs: first, species specific cell culture protocols and non-transferability of cell culture conditions between primate species and, second, inhomogeneous distribution of the differentiating NPCs leading to non-optimal survival conditions or hampered differentiation effects due to local concentration of cells and autocrine and paracrine signalling leading to challenges in phenotypic assessment of drug effects.

Most notably, differentiated primate NCs are innate sensitive to cell culture conditions and do not tolerate harsh treatment which is an inherent obstacle to producing standardized assays with these cells.

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