IN VITRO ASSAY SYSTEMS AND METHODS FOR IDENTIFYING ANTI-CANCER AGENTS

摘要

The present disclosure relates generally to drug discovery and development and, more particularly, to in vitro assay systems and methods for selecting lead anti-cancer agents for subsequent testing in human and non-human subjects. The present disclosure allows filtering for candidate anti-cancer agents that are not inactivated by liver enzymes, are able to diffuse through cell layers, are not toxic to bone marrow cells, retain anti-cancer activity in the context of stromal support, and are effective after time-limited exposure mimicking non-hepatic clearance by kidneys and other mechanisms.

主权项

1. An in vitro assay system for identifying a candidate anti-cancer agent, the system comprising:
(i) a first well having an interior volume defined by a bottom surface and at least one side wall extending from the bottom surface to an open upper end, the first well comprising:
a liver-derived layer that contains a test anti-cancer agent and is in direct contact with the bottom surface;one or more bone marrow cells adjacent the liver-derived layer; anda volume of cell culture media; (ii) a removable insert suspended within the interior volume of the first well and immersed in the volume of cell culture media, the removable insert comprising:
a lower surface comprising a semi-permeable membrane and at least one side wall extending from the lower surface to define an insert chamber;at least one tumor cell located in the insert chamber and being configured to express a detectable marker; andat least one support cell located in the insert chamber; and (iii) a second well having an interior volume defined by a bottom surface and at least one side wall extending from the bottom surface to an open upper end, the second well comprising:
a liver-derived layer that is free of the test anti-cancer agent and is in direct contact with the bottom surface;one or more bone marrow cells adjacent the liver-derived layer; anda volume of cell culture media.