Method of identifying negative hormone and/or antagonist activities

#This application claims the benefit of priority under 35 U.S.C. .sctn. 119(e) of the three following U.S. applications, each of which was filed as a nonprovisional application and converted to a provisional application by separate petitions filed on Jan. 31, 1996: Application No. 08/522,778, filed Sep. 1, 1995; application No. 08/522,779, filed Sep. 1, 1995; and application No. 08/542,648, filed Oct. 13, 1995. The complete disclosures of these related applications is hereby incorporated herein by this reference thereto.

#The present invention relates to novel compounds having retinoid negative hormone and/or retinoid antagonist-like biological activities. More specifically, the invention relates to 4-aryl substituted benzopyran, 4-aryl substituted benzothiopyran, 4-aryl substituted 1,2-dihydroquinoline and 8-aryl substituted 5,6-dihydronaphthalene derivatives which may also be substituted by a substituted 3-oxo-1-propenyl group. These novel compounds have retinoid antagonist like-activity and are useful for treating or preventing retinoid and vitamin A and vitamin A precursor induced toxicity in mammals and as an adjunct to treatment of mammals with retinoids to prevent or ameliorate unwanted or undesired side effects. The invention further relates to the use of retinoid negative hormones for increasing the biological activities of other retinoids and steroid hormones and inhibiting the basal activity of unliganded retinoic acid receptors.

#Compounds which have retinoid-like activity are well known in the art, and are described in numerous United States and other patents and in scientific publications. It is generally known and accepted in the art that retinoid-like activity is useful for treating mammals, including humans, in order to cure or alleviate the symptoms associated with numerous diseases and conditions.

#FIG. 1 shows the chemical structure of AGN 193109.

#FIGS. 2A-2F are a series of line graphs showing that AGN 193109 inhibited ATRA-dependent transactivation at the RARs. FIGS. 2A and 2B represent activity at the RAR-.alpha. receptor; FIGS. 2C and 2D represent activity at the RAR-.beta. receptor; FIGS. 2E and 2F represent activity at the RAR-.gamma. receptor. In FIGS. 2A, 2C and 2E, open squares represent retinoic acid treatment and filled circles represent AGN 193109 treatment. In FIGS. 2B, 2D and 2F the single lines represent luciferase activity measured after treatment with 10.sup.-8 M ATRA and variable concentrations of AGN 193109.

#FIGS. 3A and 3B are line graphs representing luciferase activity detected in CV-1 cells transfected with reporter plasmid ERE-tk-Luc and expression plasmid ER-RAR-.alpha. and stimulated with ATRA (FIG. 3A) or AGN 193109 (FIG. 3B) at various concentrations. Data points represent the mean .+-.SEM of three independent luciferase determinations. The results of transfections carried out using different amounts of co-transfected ER-RAR-.alpha. (0.05, 0.1 and 0.2 .mu.g/well) are indicated in each figure.

#Definitions

#For the purposes of the present invention, an RAR antagonist is defined as a chemical that binds to one or more of the RAR subtypes with a K.sub.d of less than 1 micromolar (K.sub.d <1 .mu.M) but which does not cause significant transcriptional activation of that RAR subtypes-regulated genes in a receptor co-transfection assay. Conventionally, antagonists are chemical agents that inhibit the activities of agonists. Thus, the activity of a receptor antagonist is conventionally measured by virtue of its ability to inhibit the activity of an agonist.

#An RAR agonist is defined as a chemical that binds to one or more RAR receptor subtype with K.sub.d of less than 1 micromol (K.sub.d <1 .mu.M) and causes transcriptional activation of that RAR-subtype-regulated genes in a receptor co-transfection assay. The term "RAR agonist" includes chemicals that may bind and/or activate other receptors in addition to RARs, e.g., RXR receptors.