Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1994-04-06
1996-11-26
Saunders, David
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 71, 435 29, 436500, 436503, C12Q 168, G01N 33567
Patent
active
055784453
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to an in vitro method of evaluating the effects of a substance, especially the antagonistic versus agonistic effects of a receptor-binding substance, on a selected type of cells containing endogenous intra-cellular hormone receptors.
BACKGROUND
Efforts are made in the medical industry to reduce animal tests for the evaluation of the effects of substances which are candidates for incorporation into medicines. The reasons for this is i.a. the high costs and the comparatively long time needed for the experiments. Moreover, the extrapolation of the results from animal studies to humans does not always reflect the actual effects in humans.
There is an obvious need for screening methods which give species and tissue specific, comparatively reliable prediction of the effects of drug candidates.
In cases where the mechanism of action of drug candidates is known at the cellular level, it is possible to study the effects of the candidates in vitro. In such cases it should be possible to study the effects on cell lines derived from different types of tissues, and thus get a pattern of the effects in the studied types of tissues.
The present invention provides a tool for the prediction of the antagonistic versus agonistic effects of a receptor-binding substance on different kinds of tissues in a selected species.
DESCRIPTION OF THE INVENTION
The present invention is directed to an in vitro method of evaluating the antagonistic versus agonistic effects of a receptor-binding test substance on a selected type of cells containing endogenous intra-cellular hormone receptors. The method is carried out so is distributed into several separate culture containers, such as microtiter wells, controlled chamber for an appropriate time for the establishment of stable cell growth, hormone-depleted second medium, d.sub.1 to d.sub.4, each comprising at least one container, d.sup.1 to d.sup.4, respectively, and each container being treated in the subsequent steps, known concentration, an agonist, dissolved in a second solvent, at a concentration known to result in a distinct cellular response selected to be analyzed, the same concentration as used for d.sup.1, and said reference substance, dissolved in said second solvent, at the same concentration as used for d.sup.2, the amount of the first solvent and the amount of the second solvent not exceeding a level known to be harmful to the cells, temperature and humidity controlled chamber for a period of time sufficient for the substances to affect the cells to such a degree that a distinct cellular response selected to be analyzed is reached, magnitude of the selected cellular response resulting from hormone/receptor interaction, and said selected type of cells are evaluated from a comparison of the analyzed magnitudes of the selected cellular response obtained for said groups d.sub.1 to d.sub.4.
Thus, the method is directed to the evaluation of a test substance which is known to bind to intra-cellular hormone receptors. There are two possibilities, namely that the effects of the test substance are unknown or that the properties of the intra-cellular hormone receptors are unknown. The binding of the test substance to the intra-cellular hormone receptors of a selective type of cells may be confirmed by well-known methods in the art, e.g. binding studies.
The expression "cells containing endogenous intra-cellular hormone receptors" is intended to mean that intra-cellular hormone receptors are encoded by the unmanipulated genome of the cells, contrary to the case where the genes for the hormone receptors have been transfected into cells.
In one embodiment of the invention the first solvent and the second solvent used are both added to each of the containers d.sup.1 to d.sup.4, in the amounts used for the containers of the group d.sub.1 and the group d.sub.2, respectively. In this case the possible effect of different amounts of solvents in the containers are eliminated.
In another embodiment of the invention the cells of the selected type are anchorage dep
REFERENCES:
patent: 4859585 (1989-08-01), Sonnenschein et al.
patent: 5266464 (1993-11-01), Housey
patent: 5298429 (1994-03-01), Evans et al.
Breast Cancer Research and Treatment, vol. 17, 1990, Richard Poulin, et al.: "Multiple actions of synthetic progestins on the growth of ZR-75-1 human breast cancer cells: An in vitro model for the simultaneous assay of androgen, progestin, estrogen, and glucocorticoid agonistic and antagonistic activities of . . . ".
Neuroendocrinology, vol. 50, 1989, Steven M. Gabriel, et al., "Iso Stimulation of GH and cAMP: Comparison of beta-Adrenergic-to GRF-Stimulated GH Release and cAMP Accumulation in Monolayer Cultures of Anterior Pituitary Cells in vitro", pp. 170-176, see especially p. 171.
Heeroma Karin
Nilsson Stefan
Osterlund M.ang.rten
Karo Bio Aktiebolag
Saunders David
LandOfFree
In vitro method of evaluating the effects of a substance does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with In vitro method of evaluating the effects of a substance, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and In vitro method of evaluating the effects of a substance will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1972101