Claims for Patent: 5,639,592

Summary for Patent: 5,639,592

Title:	Functional antagonism between proto-oncoprotein c-Jun and hormone receptors
Abstract:	Hormone receptors and the transcription factor Jun/AP-1 have been shown to reciprocally repress one another by a mechanism which is independent of DNA binding. For example, over-expression of AP-1 represses glucocorticoid-induced activation of genes carrying a functional glucocorticoid response element. Conversely, glucocorticoid has been shown to repress the transcriptional activation of genes which are controlled by promoters which contain the AP-1 binding site. In addition, methods are disclosed for selecting compounds useful for treating cells undergoing uncontrolled proliferation, such compounds being capable of disrupting the function of AP-1, but display substantially no ability to promote the transcriptional activation of hormone responsive genes.
Inventor(s):	Evans; Ronald M. (La Jolla, CA), Schule; Ronald (Schopfheim, DE)
Assignee:	The Salk Institute for Biological Studies (La Jolla, CA)
Application Number:	08/030,330
Patent Claims:	1. A method for selecting a compound, said method comprising selecting a compound which disrupts the function of Activator Protein-1 (AP-1), as determined by a first assay system, but does not promote transcriptional activation of a steroid hormone responsive gene, as determined by a second assay system, wherein: (a) said first assay system comprises a suitable growth medium and a cell line that expresses, in said suitable growth medium: (i) asteroid hormone receptor, (ii) AP-1, and (iii) an AP-1-responsive reporter; (b) said determination by said first assay system comprises identifying a test compound which decreases expression of said AP-1-responsive reporter when said test compound is incubated in said first assay system, thereby identifying a compound which disrupts the function of AP-1; (c) said second assay system comprises a second suitable growth medium and a cell line that expresses, in said second suitable growth medium: (i) asteroid hormone receptor, (ii) asteroid hormone-responsive reporter; and (d) said determination by said second assay system comprises identifying a test compound which does mot increase expression of said steroid hormone-responsive reporter when said test compound is incubated in said second assay system, thereby identifying a compound which does not promote transcriptional activation of asteroid hormone-responsive gene. 2. The method according to claim 1 wherein said compound forms a first complex with asteroid hormone receptor; wherein said first complex, in the presence of AP-1, disrupts the function of AP-1; and wherein said first complex is substantially unable to promote transcriptional activation of steroid hormone responsive genes. 3. The method according to claim 1 wherein said receptor is a glucocorticoid receptor, a retinoic acid receptor, a vitamin D.sub.3 receptor, a thyroid receptor, a mineralocorticoid receptor, an estrogen receptor, an estrogen-related receptor, a retinoid receptor, an androgen receptor, or a progesterone receptor. 4. A method for identifying a compound which disrupts an Activator Protein-1 (AP-1) response pathway, but which exerts no substantial effect on asteroid hormone response pathway, said method comprising identifying a compound which has both an inhibitory effect on AP-1-responsive expression, as determined by a first assay system, and no substantial effect on steroid hormone-responsive expression, as determined by a second assay system, wherein: (a) said first assay system comprises a suitable growth medium and a cell line that expresses, in said suitable growth medium: (i) asteroid hormone receptor, (ii) AP-1, and (iii) an AP-1-responsive reporter; (b) said determination by said first assay system comprises identifying a test compound which decreases expression of said AP-1-responsive reporter when said test compound is incubated in said first assay system, thereby identifying a compound which inhibits AP-1-responsive expression; (c) said second assay system comprises a second suitable growth medium and a cell line that expresses, in said second suitable growth medium: (i) asteroid hormone receptor, (ii) asteroid hormone-responsive reporter; and (d) said determination by said second assay system comprises identifying a test compound which does not increase expression of said steroid hormone-responsive reporter when said test compound is incubated in said second assay system, thereby identifying a compound which has no substantial effect on steroid hormone-responsive expression. 5. The method according to claim 4 wherein said receptor is a glucocorticoid receptor, a retinoic acid receptor, a vitamin D.sub.3 receptor, a thyroid receptor, a mineralocorticoid receptor, an estrogen receptor, an estrogen-related receptor, a retinoid receptor, an androgen receptor or a progesterone receptor. 6. A method to repress transcription activation of a steroid hormone-responsive gene by steroid hormones in an expression system that expresses a steroid hormone-responsive gene, said method comprising: exposing said system to compounds or conditions which induce Activator Protein-1 (AP-1) expression, wherein said AP-1 expression represses expression of said steroid hormone-responsive gene. 7. The method according to claim 6 wherein said steroid hormone-responsive gene is glucocorticoid-responsive, retinoic acid-responsive, vitamin D.sub.3 -responsive, thyroid hormone responsive, mineralocorticoid responsive, estrogen responsive, estrogen-related hormone responsive, androgen-responsive, progesterone-responsive, or retinoid-responsive. 8. The method according to claim 6 wherein said steroid hormone-responsive gene is glucocorticoid-responsive, thyroid hormone responsive, mineralocorticoid responsive, estrogen responsive, estrogen-related hormone responsive, androgen-responsive, progesterone-responsive, or retinoid-responsive. 9. The method according to claim 6 wherein said compounds or conditions which induce expression of AP-1 are compounds which induce tumor formation, growth factors, cytokines, neuropeptides, neurotransmitters, protein kinase c, or compounds which induce protein kinase c; or conditions of ultraviolet irradiation, gamma irradiation, stress or heat shock. 10. A method to repress transcription activation of a steroid hormone-responsive gene by steroid hormone compounds in an expression system that expresses a steroid hormone-responsive gene, said method comprising: administering to said system a peptide comprising the leucine zipper region of c-Jun in an amount effective to repress expression of said steroid hormone-responsive gene. 11. The method according to claim 10 wherein the molar ratio of the leucine zipper region of c-Jun to steroid hormone receptor falls in the range of about 0.5:1 up to 100:1. 12. A method to repress transcription activation of an Activator Protein-1 (AP-1)-responsive gene by AP-1 in an expression system that expresses an AP-1-responsive gene, said method comprising: administering to said system: (a) a composition comprising: (i) functional ligand-binding domain of steroid hormone receptor, and (ii) functional DNA-binding domain of steroid hormone receptor, and (b) a compound that binds to said ligand binding domain, in an amount effective to repress expression of said AP-1-responsive gene. 13. The method according to claim 12 wherein said AP-1-responsive gene is a collagenase gene, a c-Jun gene, a c-Fos gene, an immune-response gene, or a retinoic acid receptor-alpha gene. 14. The method according to claim 12 wherein said composition comprises functional domains of asteroid hormone receptor selected from the group consisting of a glucocorticoid receptor, a retinoic acid receptor, a vitamin D.sub.3 receptor, a thyroid receptor, a mineralocorticoid receptor, an estrogen receptor, an estrogen-related receptor, a retinoid receptor, an androgen receptor and a progesterone receptor. 15. The method according to claim 12 wherein said composition comprises functional domains of asteroid hormone receptor selected from the group consisting of a glucocorticoid receptor, a thyroid receptor, a mineralocorticoid receptor, an estrogen receptor, an estrogen-related receptor, a retinoid receptor, an androgen receptor and a progesterone receptor. 16. The method according to claim 12 wherein the molar ratio, with respect to AP-1, of each of said ligand-binding domain or said DNA-binding domain in the composition falls in the range of about 0.5:1 up to 100:1. 17. A method to overcome the repression of expression of a gene product from a gene in an expression system having a steroid hormone present, when said gene is subject to negative regulation by steroid hormone receptors, said method comprising: exposing said system to a compound or condition which induces Activator Protein-1 (AP-1) expression, wherein said AP-1 expression suppresses the repression of expression of said gene product. 18. The method according to claim 17 wherein said gene is a pro-opiomelanocortin gene, a prolactin gene, a proliferin gene, a chorionic gonadotropin alpha-subunit gene, a phosphoenolpyruvate carboxykinase gene, or a collagenase gene. 19. A method to overcome the inhibition of proliferation of cultured lymphoid cells by asteroid hormone in the presence of asteroid hormone receptor, said method comprising: exposing said lymphoid cells to compounds or conditions which induce Activator Protein-1 (AP-1) expression, wherein said AP-1 expression suppresses the inhibition of proliferation of said lymphoid cells. 20. A method for selecting a compound which disrupts the function of Activator Protein-1 (AP-1), said method comprising: 1) incubating a test compound in an assay system comprising a suitable growth medium and a cell line that expresses, in said suitable growth medium: (i) steroid hormone receptor, (ii) AP-1, and (iii) AP-1-responsive reporter; 2) detecting AP-1 responsive reporter expression; and 3) selecting a compound which decreases expression of said AP-1 responsive reporter, thereby selecting a compound which disrupts the function of AP-1. 21. A method for selecting a compound which disrupts the function of Activator Protein-1 (AP-1), but does not affect the transcriptional activation of steroid hormone-responsive genes, said method comprising: 1) incubating a test compound which disrupts the function of AP-1 in an assay system comprising a suitable growth medium and a cell line that expresses, in said suitable growth medium: (i) steroid hormone receptor, and (ii) steroid hormone-responsive reporter; and 2) selecting a test compound which does not affect the transcriptional activation of steroid hormone-responsive genes, thereby selecting a compound which disrupts the function of Activator-Protein-1 (AP-1), but does not affect the transcriptional activation of steroid hormone-responsive genes. 22. A method to repress transcription activation of an Activator Protein-1 (AP-1)-responsive gene by AP-1 in an expression system that expresses an AP-1-responsive gone, said method comprising: administering to said system: (a) a composition comprising: (i) functional ligand-binding domain of steroid hormone receptor, and (ii) functional DNA-binding domain of steroid hormone receptor, in an amount effective to repress expression of said AP-1-responsive gene, wherein said system comprises an endogenous compound that binds to said ligand binding domain.

Details for Patent 5,639,592

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	01/15/1974		2014-06-17
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	12/27/1984		2014-06-17
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	02/15/1985		2014-06-17
Ferring Pharmaceuticals Inc.	NOVAREL	chorionic gonadotropin	For Injection	017016	02/16/1990		2014-06-17
Bel-mar Laboratories, Inc.	CHORIONIC GONADOTROPIN	chorionic gonadotropin	Injection	017054	03/26/1974		2014-06-17
Fresenius Kabi Usa, Llc	CHORIONIC GONADOTROPIN	chorionic gonadotropin	For Injection	017067	03/05/1973		2014-06-17
Smith & Nephew, Inc.	SANTYL	collagenase	Ointment	101995	06/04/1965		2014-06-17