WO2016065177A1 - Method of treating depression and other stress related disorders - Google Patents

Abstract

The present invention includes a method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFrl antagonist, wherein the disorder manifests hypersecretion of corticotropin-releasing factor (CRF). The method further comprising administering to the male subject a pharmaceutically effective amount of a second therapeutic agent.

Description

TITLE OF THE INVENTION

Method of Treating Depression and Other Stress Related Disorders CROSS-REFERENCE TO RELATED APPLICATIONS The present application claims priority to U.S. Provisional Patent Application No.62/067,204, filed October 22, 2014, which application is incorporated herein by reference in its entirety. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under grant number NIH MH073030 awarded by the National Institute of Health. The government has certain rights in the invention. BACKGROUND OF THE INVENTION

Corticotropin-releasing factor (CRF) represents an important link between stress and mood regulation. Studies have suggested that stress induced elevations in CRF contribute to neuropsychiatric disease development through excessive activation of its type 1 receptor, corticotropin-releasing factor receptor-1 (CRFr1). Consequently, CRFr1 has received considerable attention as a novel pharmaceutical target for the treatment of stress- related affective disorders. GlaxoSmithKline, Pfizer, Neurocrine Biosciences,

DuPont/Bristol-Myers Squibb, and others have developed CRFr1 small molecule antagonists toward this end. However, despite compelling results for antidepressant-like and anxiolytic- like effects of these drugs in pre-clinical studies in rodents and nonhuman primates, none of the CRFr1 antagonists brought to clinical trial over the past decade have successfully completed a Phase III trial (Koob, et al., Neuropsychopharmacology 2012, 37:308–309).

Considerable evidence supports the involvement of CRFr1 in stress modulation of the serotonergic (5-HTergic) dorsal raphe nucleus (DR) in regulation of mood and affect. A disruption in the ability of CRF to regulate 5-HT circuits during chronic stress is implicated in affective disorder pathophysiology.

There is a need in the art for novel methods of treating stress or anxiety in a mammal in need thereof. The present invention fulfills this need. BRIEF SUMMARY OF THE INVENTION

In one aspect, the present invention includes a method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist, or a salt or solvate thereof, wherein the disorder manifests hypersecretion of corticotropin-releasing factor (CRF) and the disorder is selected from the group consisting of an anxiety-related disorder, a mood disorder, a post-traumatic stress disorder, immune suppression, drug or alcohol withdrawal symptoms, a sleep disorder induced by stress, fibromyalgia, dysthemia, a bipolar disorder, cyclothymia, fatigue syndrome, stress-induced irritable bowel syndrome, and stress-induced headache.

In certain embodiments, the disorder is selected from the group consisting of an anxiety-related disorder, a mood disorder, a bipolar disorder, and a post-traumatic stress disorder.

In other embodiments, the anxiety-related disorder is selected from the group consisting of an anxiety state, a generalized anxiety disorder, a social anxiety disorder, an anxiety with co-morbid depressive illness, a panic disorder, an obsessive-compulsive disorder, a phobic disorder, a post-traumatic stress disorder, and an atypical anxiety disorder.

In yet other embodiments, the mood disorder is depression selected from the group consisting of major depression, single episode depression, recurrent depression, child abuse induced depression, and postpartum depression.

In yet other embodiments, the CRFr1 antagonist is selected from the group consisting of LY2371712, NBI-35965, NBI-30775, Antalarmin, CP-316,311, CRA 5626, CP- 154,526, Emicerfont, ONO-2333Ms, Pexacerfont, SSR125543, NBI-34101 , DMP-696, DMP-904, DMP-695, SC-241, BMS-561388, R121919, NBI-30545, CP-376,396, NBI- 27914, NBI-34101, PF-572778, GSK561579, and GSK586529.

In another aspect, the present invention includes a method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist, or a salt or solvate thereof, and a second therapeutic agent, or a salt or solvate thereof, wherein the disorder manifests hypersecretion of corticotropin-releasing factor (CRF) and the second therapeutic agent is selected from the group consisting of sertraline, fluoxetine, citalopram, escitalopram, paroxetine, fluvoxamine, trazodone, desvenlafaxine , duloxetine, venlafaxine, amitriptyline, amoxapine, clomipramine, desipramine, doxepin, imipramine, nortriptyline, protriptyline, trimipramine, maprotiline, nefazodone, trazodone, isocarboxazid, phenelzine, selegiline, tranylcypromine, mirtazapine, quetiapene, buspirone, bupropion, a selective serotonin reuptake inhibitor, a serotonin and norepinephrine reuptake inhibitor, a tricyclic antidepressant, a tetracyclic antidepressant, a dopamine reuptake blocker, a 5-HT2 receptor antagonist, a 5-HT3 receptor antagonist, a monoamine oxidase inhibitor, a noradrenergic antagonist, and a mixture thereof.

In one embodiment, the male subject is a male human. In another embodiment, the CRFr1 antagonist is simultaneously administered with the second therapeutic agent. In yet another embodiment, the CRFr1 antagonist is administered 1-6 hours before the administration of the second therapeutic agent. In yet another embodiment, the CRFr1 antagonist is administered 1 -6 hours after the administration of the second therapeutic agent.

BRIEF DESCRIPTION OF THE DRAWINGS

For the purpose of illustrating the invention, there are depicted in the drawings certain embodiments of the invention. However, the invention is not limited to the precise arrangements and instrumentalities of the embodiments depicted in the drawings.

FIG. 1 illustrates the effects of small molecule antagonist NBI 35965 and or corticotropin-releasing factor (CRF) on dorsal raphe infusion of the corticotropin-releasing factor receptor-1 (CRFr1 ). Panel A illustrates the time course of corticosterone response to a 15-min restraint (indicated by shaded region) in males. Panel B illustrates the area under the curve analysis demonstrating a significant reduction of corticosterone output by NBI 35965 in males. Panel C illustrates the time course of corticosterone response to a 15-min restraint (indicated by shaded region) in females. Panel D illustrates the area under the curve analysis demonstrating not a significant reduction of corticosterone output by NBI 35965 in females. Panel E illustrates corticosterone response to infusion of 50 ng CRF in males. Panel F illustrates the area under the curve analysis demonstrating that CRF enhancement of corticosterone release was specific to males. Panel G illustrates the corticosterone response to infusion of 50 ng CRF in females. Panel H illustrates the area under the curve analysis demonstrating that CRF enhancement of corticosterone release was not significant to females. Data are presented as mean values ± SEM (n = 8). *p <

.05 in comparison with artificial cerebrospinal fluid (ACSF).

FIG. 2 illustrates the finding that males and females show divergent behavioral responses to CRFr1 antagonism or CRF infusion into the dorsal raphe. Panel A illustrates the effects of CRFr-1 antagonist NBI 35965 on total immobile time. Panel B illustrates latency to first bout of immobility in the tail suspension test (TST). Males showed a greater latency to immobility. Panel C illustrates the finding that CRFr1 antagonism had no effect in the light-dark box (LD) on measures of total time spent in the light compartment. Panel D illustrates the finding that CRFr1 antagonists had no effect in latency to first exit from the light compartment. Panel E illustrates the finding that the 50-ng CRF infusion reduced immobile time. Panel F illustrates the finding that the 50-ng CRF infusion increased latency to immobility in males in the TST. Panel G illustrates the finding that the 50-ng CRF infusion increased total light time in males in the LD. Panel H illustrates the finding that the 50-ng CRF infusion increased latency to exit the light in males in the LD. Data are presented as mean values ± SEM (n = 8). *p <0.05 in comparison with artificial cerebrospinal fluid (ACSF).

FIG. 3 illustrates the finding that a 50-ng corticotropin-releasing factor (CRF) infusion elicited differential patterns of neuronal activation in males and females across dorsal raphe (DR) subregions. Panels A and B illustrate the finding that cFos induction 90 min after CRF infusion is enhanced in males shown by representative dorsomedial (dmDR) sections. Panels C and D illustrate the finding that cFos induction 90 min after CRF infusion is reduced in females shown by representative dorsomedial (dmDR) sections. Panel E illustrates the finding that Fos-positive cell counts from DR subregions demonstrate increased counts in males but reduced counts in females, particularly in dmDR and rostral lateral wing (rLW) and caudal lateral wing (cLW) subregions. Values represent the difference in mean number of Fos-positive cells in sections from mice administered CRF or artificial cerebrospinal fluid (ASCF). Data represent the difference in averages from 7–9 mice/group. *p<0.05.

FIG. 4 illustrates the finding that CRFr1 is expressed on different cell populations

in males and females. Panel A illustrates expression of genes relevant to CRF and serotonergic signaling in the DR. There were no sex differences in messenger RNA levels of CRFr1, CRFr2, CRF-binding protein (CRF-BP), γ-aminobutyric acid receptor (GABR) subunits α1 , α2, δ, or γ2; however, tryptophan hydroxylase (TPH)2 was significantly higher in females. Data are presented as fold change relative to the mean value in males ±SEM (n =5); *p <0.05 compared with males. Panel B illustrates regional but no sex differences in number of green fluorescent protein immunoreactive (GFP-ir) cells throughout subregions of the DR. Panel C illustrates brain atlas diagrams depict the region viewed in the

representative images of the rLW. Panel C illustrates brain atlas diagrams depicting the region viewed in the representative images of the dmDR. Panels E and G illustrate representative, split channel confocal images from the rLW. Panels I and K illustrate representative, split channel confocal images from the dmDR. Panels F and G illustrate sections dual-labeled with anti-parvalbumin (PV) to identify the phenotype of GFP cells in the DR. Panels J and K illustrate sections dual-labeled with anti-TPH to identify the phenotype of GFP cells in the DR. Arrows denote location of GFP-ir cells enlarged to illustrate co-localization (single arrow, GFP only; double arrow, co-localized). Panel M illustrates probability that a given GFP-ir cell colocalizes with PV. Panel N illustrates probability that a given GFP-ir cell colocalizes with TPH. Females presented with an overall reduction in the probability that a given GFP-ir cell colocalizes with PV and reduced likelihood that GFP-ir cells within the rDR colocalize with TPH. Data are presented as an average probability ±SEM. *p<0.05 in comparison with male.

FIG. 5 illustrates the finding that males and females exhibit differences in baseline characteristics as well as divergent responses to bath applied corticotropin-releasing factor (CRF). Panel A illustrates representative inhibitory postsynaptic current trace. Panel B illustrates scaled inhibitory postsynaptic currents averaged from a single male. Panel C illustrates scaled inhibitory postsynaptic currents averaged from a single female. Insets B’ and C’ illustrate sex difference in effect of CRF on rise time. Panel D illustrates the finding that CRF rise time increases in males but decreases in females. Panel E illustrates the finding that CRF half-width increases in males but decreases in females. Panel F illustrates the finding that CRF decay time increases in males but decreases in females. Panel G illustrates the finding that males exhibit increased excitability compared with females to a series of current injections. Values represent the difference in response of CRF–DNQX (n = 7–9).*p <0.05. DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the unexpected discovery of gender or sex differences in behavioral and physiological effects of a CRFr1 antagonist and CRF in the DR.

Research has identified that gender differences are mechanistically associated in receptor co-expression and divergent electrophysiological responses to CRF in 5-HTergic neurons. The blunted response of females points to a potential explanation for the lack of efficacy in CRFr1 antagonists in clinical trials, which have focused primarily on female participants due to their increased disease prevalence (Table 3). These findings support the importance of identifying sex differences in central stress pathways to understand the heightened predisposition of females toward these disorders and in identifying sex appropriate and potentially sex-specific pharmacological treatments.

In one aspect, the invention includes a method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist or a salt or solvate thereof, wherein the disorder manifests hypersecretion of corticotropin-releasing factor (CRF).

In another aspect, the invention includes a method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist or a salt or solvate thereof, together with a second therapeutic agent or a salt or solvate thereof. Definitions

As used herein, each of the following terms has the meaning associated with it in this section. Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and pharmacology are those well-known and commonly employed in the art.

As used herein, the articles“a” and“an” refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example,“an element” means one element or more than one element.

As used herein, the term“about” is understood by persons of ordinary skill in the art and varies to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, or ±0.l% from the specified value, as such variations are appropriate to perform the disclosed methods.

As used herein, the term“CNS” refers to central nervous system. “Instructional material,” as that term is used herein, includes a publication, a recording, a diagram, or any other medium of expression that may be used to communicate the usefulness of the compounds and/or compositions of the invention. In some instances, the instructional material may be part of a kit useful for effecting alleviating or treating the various diseases or disorders recited herein. Optionally, or alternately, the instructional material may describe one or more methods of alleviating the diseases or disorders in a cell or a tissue of a mammal. The instructional material of the kit may, for example, be affixed to a container that contains the compounds and/or compositions of the invention or be shipped together with a container that contains the compounds and/or compositions. Alternatively, the instructional material may be shipped separately from the container with the intention that the recipient uses the instructional material and the compound and/or composition

cooperatively. For example, the instructional material is for use of a kit; instructions for use of the compound and/or composition; or instructions for use of a formulation of the compound and/or composition.

As used herein, the term“patient” or“individual” or“subject” refers to a human or a non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. In certain embodiments, the patient, individual or subject is human.

As used herein, the terms“pharmaceutically effective amount” and“effective amount” and“therapeutically effective amount” refer interchangeably to a nontoxic but sufficient amount of an agent to provide the desired biological result. That result may be reduction and/or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. An appropriate therapeutic amount in any individual case may be determined by one of ordinary skill in the art using routine

experimentation.

As used herein, the term“pharmaceutically acceptable” refers to a material, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound, and is relatively non-toxic, i.e., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

As used herein, the term“pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the invention within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be“acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the patient. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil;

glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid;

pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein,“pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the invention, and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. The“pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound useful within the invention. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the invention are known in the art and described, for example in Remington’s Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

As used herein, the language“pharmaceutically acceptable salt” refers to a salt of the administered compound prepared from pharmaceutically acceptable non-toxic acids and bases, including inorganic acids, inorganic bases, organic acids, inorganic bases, solvates, hydrates, and clathrates thereof. Suitable pharmaceutically acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of inorganic acids include sulfate, hydrogen sulfate, hydrochloric, hydrobromic, hydriodic, nitric, carbonic, sulfuric, and phosphoric acids (including hydrogen phosphate and dihydrogen phosphate). Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic,

ethanesulfonic, benzenesulfonic, pantothenic, trifluoromethanesulfonic, 2- hydroxyethanesulfonic, p-toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, alginic, β-hydroxybutyric, salicylic, galactaric and galacturonic acid. Suitable

pharmaceutically acceptable base addition salts of compounds of the invention include, for example, ammonium and metallic salts including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include organic salts made from basic amines such as, for example, N,N’-dibenzylethylene-diamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts may be prepared from the corresponding compound by reacting, for example, the appropriate acid or base with the compound.

As used herein, the term“pharmaceutical composition” or“composition” refers to a mixture of at least one compound useful within the invention with a

pharmaceutically acceptable carrier. The pharmaceutical composition facilitates

administration of the compound to a patient. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, inhalational, rectal, vaginal, transdermal, intranasal, buccal, sublingual, parenteral, intrathecal, intragastrical, ophthalmic, pulmonary and topical administration.

As used herein, the term“prevent” or“prevention” means no disorder or disease development if none had occurred, or no further disorder or disease development if there had already been development of the disorder or disease. Also considered is the ability of one to prevent some or all of the symptoms associated with the disorder or disease.

As used herein, the term“solvate” refers to a complex with one or more solvent molecules, which may comprise water, methanol, ethanol, 1-propanol, 2-propanol, DMSO, DMF, ethyl ether, acetone, and/or MTBE, and the like.

As used herein, the term“substrate” as relating to a drug efflux transporter refers to a compound (such as a small molecule compound, peptide or protein) that is transported across an extra- or intracellular membrane by the drug efflux transporter.

As used herein, the term“treatment” or“treating” is defined as the application or administration of a therapeutic agent, i.e., a compound useful within the invention (alone or in combination with another pharmaceutical agent), to a patient, or application or administration of a therapeutic agent to an isolated tissue or cell line from a patient (e.g., for diagnosis or ex vivo applications), who has a disease or disorder, a symptom of a disease or disorder or the potential to develop a disease or disorder, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease or disorder, the symptoms of the disease or disorder, or the potential to develop the disease or disorder. Such treatments may be specifically tailored or modified, based on knowledge obtained from the field of pharmacogenomics.

As used herein, the term“sertraline” refers to (1S,4S)-4-(3,4-dichlorophenyl)- N-methyl-1 ,2,3,4-tetrahydronaphthalen-1-amine, or a salt or solvate thereof.

As used herein, the term“fluoxetine” refers to (±)-N-methyl-3-phenyl-3- [(α,α,α-trifluoro-p-tolyl)oxy]propylamine, or a salt or solvate thereof.

As used herein, the term“citalopram” refers to (RS)-1-[3-(dimethylamino)- propyl]-1-(4-fluorophenyl)-1 ,3-dihydroisobenzofuran-5-carbonitrile, or a salt or solvate thereof.

As used herein, the term“escitalopram” refers to (S)-1-[3-(dimethylamino)- propyl]-1-(4-fluorophenyl)-1 ,3-dihydroisobenzofuran-5-carbonitrile, or a salt or solvate thereof.

As used herein, the term“paroxetine” refers to (3S,4R)-3-[(2H-1,3- benzodioxol-5-yloxy)methyl]-4-(4-fluorophenyl)piperidine, or a salt or solvate thereof.

As used herein, the term“fluvoxamine” refers to 2-{[(E)-{5-methoxy-1-[4- (trifluoromethyl)phenyl]pentylidene}amino]oxy}ethanamine, or a salt or solvate thereof.

As used herein, the term“trazodone” refers to 2-{3-[4-(3- chlorophenyl)piperazin-1-yl]propyl}[1 ,2,4]triazolo[4,3-a]pyridin-3(2H)-one, or a salt or solvate thereof.

As used herein, the term“desvenlafaxine” refers to 4-[2-dimethylamino-1-(1- hydroxycyclohexyl) ethyl]phenol, or a salt or solvate thereof.

As used herein, the term“duloxetine” refers to (+)-(S)-N-methyl-3- (naphthalen-1-yloxy)-3-(thiophen-2-yl)propan-1-amine, or a salt or solvate thereof.

As used herein, the term“venlafaxine” refers to (RS)-1-[2-dimethylamino-1- (4-methoxyphenyl)-ethyl]cyclohexanol, or a salt or solvate thereof.

As used herein, the term“amitriptyline” refers to 3-(10,11-dihydro-5H- dibenzo[a,d]cycloheptene-5-ylidene)-N,N-dimethylpropan-1-amine, or a salt or solvate thereof.

As used herein, the term“amoxapine” refers to 2-chloro-11-(piperazin-1- yl)dibenzo[b,f][1 ,4]oxazepine, or a salt or solvate thereof.

As used herein, the term“clomipramine” refers to 3-(3-chloro-10,11-dihydro- 5H-dibenzo[b,f]azepin-5-yl)-N,N-dimethylpropan-1-amine, or a salt or solvate thereof.

As used herein, the term“desipramine” refers to 3-(10,11-dihydro-5H- dibenzo[b,f]azepin-5-yl)-N-methylpropan-1-amine, or a salt or solvate thereof.

As used herein, the term“doxepin” refers to (E/Z)-3-(dibenzo[b,e]oxepin- 11(6H)-ylidene)-N,N-dimethylpropan-1-amine, or a salt or solvate thereof.

As used herein, the term“imipramine” refers to 3-(10,11-dihydro-5H- dibenzo[b,f]azepin-5-yl)-N,N-dimethylpropan-1 -amine, or a salt or solvate thereof.

As used herein, the term“nortriptyline” refers to 3-(10,11-dihydro-5H- dibenzo[a,d]cyclohepten-5-ylidene)-N-methyl-1 -propanamine, or a salt or solvate thereof.

As used herein, the term“protriptyline” refers to 3-(5H- dibenzo[a,d][7]annulen-5-yl)-N-methylpropan-1-amine, or a salt or solvate thereof.

As used herein, the term“trimipramine” refers to (±)-3-(10,11 -dihydro-5H- dibenzo[b,f]azepin-5-yl)-N,N,2-trimethylpropan-1-amine, or a salt or solvate thereof.

As used herein, the term“maprotiline” refers to N-Methyl-9,10- ethanoanthracene-9(10H)-propanamine, or a salt or solvate thereof.

As used herein, the term“nefazodone” refers to 1 -(3-[4-(3-chlorophenyl)- piperazin-1-yl]propyl)-3-ethyl-4-(2-phenoxyethyl)-1H-1 ,2,4-triazol-5(4H)-one, or a salt or solvate thereof.

As used herein, the term“trazodone” refers to 2-{3-[4-(3- chlorophenyl)piperazin-1-yl]propyl}[1 ,2,4]triazolo[4,3-a]pyridin-3(2H)-one, or a salt or solvate thereof.

As used herein, the term“isocarboxazid” refers to N′-benzyl-5- methylisoxazole-3-carbohydrazide, or a salt or solvate thereof.

As used herein, the term“phenelzine” refers to 2-phenylethylhydrazine, or a salt or solvate thereof.

As used herein, the term“selegiline” refers to (R)-N-methyl-N-(1- phenylpropan-2-yl)prop-1-yn-3-amine, or a salt or solvate thereof.

As used herein, the term“tranylcypromine” refers to (±)-trans-2- phenylcyclopropan-1-amine or 1 R*,2S*)-2-phenylcyclopropan-1-amine, or a salt or solvate thereof.

As used herein, the term“mirtazapine” refers to (±)-2-methyl-1,2,3,4,10,14b- hexahydropyrazino[2,1-a]pyrido[2,3-c][2]benzazepine, or a salt or solvate thereof.

As used herein, the term“quetiapine” refers to 2-(2-(4-dibenzo[b,f][1,4]- thiazepine- 11-yl- 1 -piperazinyl)ethoxy)ethanol, or a salt or solvate thereof.

As used herein, the term“buspirone” refers to 8-[4-(4-pyrimidin-2- ylpiperazin-1-yl)butyl]-8-azaspiro[4.5]decane-7,9-dione, or a salt or solvate thereof.

As used herein, the term“bupropion” refers to (±)-2-(tert-Butylamino)-1-(3- chlorophenyl)propan-1-one, or a salt or solvate thereof.

As used herein, the term“NBI-30775” or“R-121919” refers to 3-[6- (dimethylamino)-4-methyl-3-pyridinyl]-2,5-dimethyl-N,N-dipropylpyrazolo[1,5-a]pyrimidin- 7-amine, or a salt or solvate thereof.

As used herein, the term“Antalarmin” refers to N-butyl-N-ethyl-2,5,6- trimethyl-7-(2,4,6-trimethylphenyl)pyrrolo[3,2-e]pyrimidin-4-amine, or a salt or solvate thereof.

As used herein, the term“ONO-2333Ms” refers to 10-(2-chloro-4- methoxyphenyl)-11 -methyl-N-(pentan-3-yl)-1,8,12-triazatricyclo[7.3.0.0^3,7]dodeca-2,7,9,11- tetraen-2-amine, or a salt or solvate thereof.

As used herein, the term“Pexacerfont” refers to 8-(6-methoxy-2- methylpyridin-3-yl)-2,7-dimethyl-N-[(1R)-1-methylpropyl]pyrazolo[1,5-a]-1,3,5-triazin-4- amine, or a salt or solvate thereof.

As used herein, the term“SSR125543” refers to (S)-4-(2-chloro-4-methoxy-5- methylphenyl)-N-(2-cyclopropyl-1-(3-fluoro-4-methylphenyl)ethyl)-5-methyl-N-(prop-2- ynyl)thiazol-2-amine, or a salt or solvate thereof.

Throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible sub-ranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.1, 5.3, 5.5, and 6. This applies regardless of the breadth of the range. Description

In one aspect, the present invention includes a method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist, or a salt or solvate thereof, wherein the disorder manifests hypersecretion of corticotropin-releasing factor (CRF).

In certain embodiments, the disorder manifests hypersecretion of CRF. In other embodiments, the disorder is selected from the group consisting of anxiety-related disorders, mood disorders, post-traumatic stress disorder, immune suppression, drug or alcohol withdrawal symptoms, sleep disorders induced by stress, fibromyalgia, dysthemia, bipolar disorders, cyclothymia, fatigue syndrome, stress-induced irritable bowel syndrome, and stress-induced headache. In yet other embodiments, wherein the disorder is selected from the group consisting of anxiety-related disorders, mood disorders, bipolar disorders, and post-traumatic stress disorder.

In certain embodiments, the anxiety-related disorder is selected from the group consisting of anxiety states, generalized anxiety disorder, social anxiety disorder, anxiety with co-morbid depressive illness, panic disorder, obsessive-compulsive disorder, phobic disorders, post-traumatic stress disorder, and atypical anxiety disorders. In other

embodiments, the mood disorder is selected from the group consisting of depression, including major depression, single episode depression, recurrent depression, child abuse induced depression, and postpartum depression; dysthemia; bipolar disorders; and cyclothymia.

The CRFr1 antagonist for the invention is selected from the group consisting of LY2371712, NBI-35965, NBI-30775 (Neurocrine), Antalarmin, CP-316,311 (Pfizer), CRA 5626, CP-154,526 (Pfizer), Emicerfont (Glaxo), ONO-2333Ms (Ono Pharmaceuticals), Pexacerfont (Bristol-Myers-Squibb), SSR125543 (Sanofi-Aentis), NBI-34101 (Neurocrine), DMP-696, DMP-904, DMP-695, SC-241, BMS-561388, R121919, NBI-30545, CP-376,396, NBI-27914, NBI-34101, PF-572778, GSK561579 and GSK586529.

The corresponding structural formulas of some of the above-mentioned CRFr1 antagonist for use in the present invention are set out in the table below.

SC-241

GSK561579

In another aspect, the present invention includes a method of treating a disorder in a male subject. The method comprises administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist or a salt or solvate thereof, and a second therapeutic agent or a salt or solvate. In certain embodiments, the second therapeutic agent is selected from the group consisting of selective serotonin reuptake inhibitors, serotonin and norepinephrine reuptake inhibitors, tricyclic antidepressants, tetracyclic antidepressants, dopamine reuptake blocker, 5-HT2 receptor antagonists, 5-HT3 receptor antagonists, monoamine oxidase inhibitors, noradrenergic antagonist, and a mixture thereof. In other embodiments, the second therapeutic agent is selected from the group consisting of sertraline, fluoxetine, citalopram, escitalopram, paroxetine, fluvoxamine, trazodone, desvenlafaxine , duloxetine, venlafaxine, amitriptyline, amoxapine, clomipramine, desipramine, doxepin, imipramine, nortriptyline, protriptyline, trimipramine, maprotiline, nefazodone, trazodone, isocarboxazid, phenelzine, selegiline, tranylcypromine, mirtazapine, quetiapine, buspirone, bupropion, and a mixture thereof.

In one embodiment, the CRFr1 antagonist is administered simultaneously with the second therapeutic agent. In another embodiment, the CRFr1 antagonist is formulated with the second therapeutic agent, then the formulated product is administered to a male subject in need thereof. In yet another embodiment, the CRFr1 antagonist is administered 1-6 hours before the administration of the second therapeutic agent. In yet another embodiment, the CRFr1 antagonist is administered 1 -6 hours after the administration of the second therapeutic agent.

In yet another aspect, the invention includes a pharmaceutical composition comprising a CRFr1 antagonist. In certain embodiments, the pharmaceutical composition further comprises a second therapeutic agent.

In certain embodiments, the male subject treated by the method of the invention is a male human. Administration/Dosage/Formulations

The regimen of administration may affect what constitutes an effective amount. The therapeutic formulations may be administered to the patient either prior to, around the time, or after the onset of a disease or disorder. Further, several divided dosages, as well as staggered dosages may be administered daily or sequentially, or the dose may be continuously infused, or may be a bolus injection. Further, the dosages of the therapeutic formulations may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.

Administration of the compositions useful within the present invention to a patient, such as a mammal, such as a human, may be carried out using known procedures, at dosages and for periods of time effective to treat a disease or disorder in the patient. An effective amount of the therapeutic compound necessary to achieve a therapeutic effect may vary according to factors such as the state of the disease or disorder in the patient; the age, sex, and weight of the patient; and the ability of the therapeutic compound to treat a disease or disorder in the patient. Dosage regimens may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. In certain embodiments, an effective dose range for a therapeutic compound of the invention ranges from about 1 to about 5,000 mg/kg of body weight/day. In other embodiments, an effective dose range for a therapeutic compound of the invention ranges from about 100 to about 1,000 mg/kg of body weight/day. In yet other embodiments, an effective dose range for a therapeutic compound of the invention ranges from about 10 to about 50 mg/kg of body weight/day. In yet other embodiments, an effective dose range for a therapeutic compound of the invention ranges from about 1 to about 50 mg/kg of body weight/day. In yet other embodiments, an effective dose range for a therapeutic compound of the invention ranges from about 1 to about 10 mg/kg of body weight/day. In yet other embodiments, an effective dose range for a therapeutic compound of the invention is about 2.5 mg/kg of body weight/day. One of ordinary skill in the art would be able to study the relevant factors and make the determination regarding the effective amount of the therapeutic compound without undue experimentation.

Actual dosage levels of the active ingredients in the pharmaceutical compositions of this invention may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

In particular, the selected dosage level will depend upon a variety of factors including the activity of the particular compound employed, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds or materials used in combination with the compound, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well, known in the medical arts.

A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

In particular embodiments, it is especially advantageous to formulate the compound in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined quantity of therapeutic compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the therapeutic compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of

compounding/formulating such a therapeutic compound for the treatment of a disease or disorder in a patient.

In certain embodiments, the compositions useful within the invention are formulated using one or more pharmaceutically acceptable excipients or carriers. In certain embodiments, the pharmaceutical compositions of the invention comprise a therapeutically effective amount of a compound useful within the invention and a pharmaceutically acceptable carrier.

The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it may include isotonic agents, for example, sugars, sodium chloride, or polyalcohols such as mannitol and sorbitol, in the composition. Prolonged absorption of the injectable compositions may be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.

In certain embodiments, the compositions useful within the invention are administered to the patient in dosages that range from one to five times per day or more. In other embodiments, the compositions useful within the invention are administered to the patient in range of dosages that include, but are not limited to, once every day, every two, days, every three days to once a week, and once every two weeks. It will be readily apparent to one skilled in the art that the frequency of administration of the various combination compositions useful within the invention will vary from individual to individual depending on many factors including, but not limited to, age, disease or disorder to be treated, gender, overall health, and other factors. Thus, the invention should not be construed to be limited to any particular dosage regime and the precise dosage and composition to be administered to any patient will be determined by the attending physical taking all other factors about the patient into account.

Compounds for administration may be in the range of from about 1 µg to about 10,000 mg, about 20 µg to about 9,500 mg, about 40 µg to about 9,000 mg, about 75 µg to about 8,500 mg, about 150 µg to about 7,500 mg, about 200 µg to about 7,000 mg, about 3050 µg to about 6,000 mg, about 500 µg to about 5,000 mg, about 750 µg to about 4,000 mg, about 1 mg to about 3,000 mg, about 10 mg to about 2,500 mg, about 20 mg to about 2,000 mg, about 25 mg to about 1,500 mg, about 50 mg to about 1,000 mg, about 75 mg to about 900 mg, about 100 mg to about 800 mg, about 250 mg to about 750 mg, about 300 mg to about 600 mg, about 400 mg to about 500 mg, and any and all whole or partial increments therebetween. In certain embodiments, the dose of a compound is from about 1 mg and about 2,500 mg. In other embodiments, a dose of a compound of the invention used in

compositions described herein is less than about 10,000 mg, or less than about 8,000 mg, or less than about 6,000 mg, or less than about 5,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in other embodiments, a dose of a second compound (i.e., a drug used for treating a disease or disorder) as described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg, or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any and all whole or partial increments thereof.

In certain embodiments, the present invention is directed to a packaged pharmaceutical composition comprising a container holding a therapeutically effective amount of a compound of the invention, alone or in combination with a second

pharmaceutical agent; and instructions for using the compound to treat, prevent, or reduce one or more symptoms of a disease or disorder in a patient.

Formulations may be employed in admixtures with conventional excipients, i.e., pharmaceutically acceptable organic or inorganic carrier substances suitable for oral, parenteral, nasal, intravenous, subcutaneous, enteral, intranasal drug delivery, or any other suitable mode of administration, known to the art. The pharmaceutical preparations may be sterilized and if desired mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure buffers, coloring, flavoring and/or aromatic substances and the like. They may also be combined where desired with other active agents, e.g., other cognition improving agents.

The term“container” includes any receptacle for holding the pharmaceutical composition. For example, in certain embodiments, the container is the packaging that contains the pharmaceutical composition. In other embodiments, the container is not the packaging that contains the pharmaceutical composition, i.e., the container is a receptacle, such as a box or vial that contains the packaged pharmaceutical composition or unpackaged pharmaceutical composition and the instructions for use of the pharmaceutical composition. Moreover, packaging techniques are well known in the art. It should be understood that the instructions for use of the pharmaceutical composition may be contained on the packaging containing the pharmaceutical composition, and as such the instructions form an increased functional relationship to the packaged product. However, it should be understood that the instructions may contain information pertaining to the compound’s ability to perform its intended function, e.g., treating, preventing, or reducing a disease or disorder in a patient.

Routes of administration of any of the compositions of the invention include oral, nasal, rectal, intravaginal, parenteral, buccal, sublingual, intranasal drug delivery, or topical. The compounds for use in the invention may be formulated for administration by any suitable route, such as for oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous,

intrabronchial, inhalation, and topical administration.

Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions that would be useful in the present invention are not limited to the particular formulations and compositions that are described herein. Oral Administration

For oral application, particularly suitable are tablets, dragees, liquids, drops, suppositories, or capsules, caplets and gelcaps. The compositions intended for oral use may be prepared according to any method known in the art and such compositions may contain one or more agents selected from the group consisting of inert, non-toxic pharmaceutically excipients which are suitable for the manufacture of tablets. Such excipients include, for example an inert diluent such as lactose; granulating and disintegrating agents such as cornstarch; binding agents such as starch; and lubricating agents such as magnesium stearate. The tablets may be uncoated or they may be coated by known techniques for elegance or to delay the release of the active ingredients. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert diluent.

For oral administration, the compounds may be in the form of tablets or capsules prepared by conventional means with pharmaceutically acceptable excipients such as binding agents (e.g., polyvinylpyrrolidone, hydroxypropylcellulose or

hydroxypropylmethylcellulose); fillers (e.g., cornstarch, lactose, microcrystalline cellulose or calcium phosphate); lubricants (e.g., magnesium stearate, talc, or silica); disintegrates (e.g., sodium starch glycollate); or wetting agents (e.g., sodium lauryl sulphate). If desired, the tablets may be coated using suitable methods and coating materials such as OPADRY™ film coating systems available from Colorcon, West Point, Pa. (e.g., OPADRY™ OY Type, OYC Type, Organic Enteric OY-P Type, Aqueous Enteric OY-A Type, OY-PM Type and

OPADRY™ White, 32K18400). Liquid preparation for oral administration may be in the form of solutions, syrups or suspensions. The liquid preparations may be prepared by conventional means with pharmaceutically acceptable additives such as suspending agents (e.g., sorbitol syrup, methyl cellulose or hydrogenated edible fats); emulsifying agent (e.g., lecithin or acacia); non-aqueous vehicles (e.g., almond oil, oily esters or ethyl alcohol); and preservatives (e.g., methyl or propyl p-hydroxy benzoates or sorbic acid).

Granulating techniques are well known in the pharmaceutical art for modifying starting powders or other particulate materials of an active ingredient. The powders are typically mixed with a binder material into larger permanent free-flowing agglomerates or granules referred to as a“granulation.” For example, solvent-using“wet” granulation processes are generally characterized in that the powders are combined with a binder material and moistened with water or an organic solvent under conditions resulting in the formation of a wet granulated mass from which the solvent must then be evaporated.

Melt granulation generally consists in the use of materials that are solid or semi-solid at room temperature (i.e. having a relatively low softening or melting point range) to promote granulation of powdered or other materials, essentially in the absence of added water or other liquid solvents. The low melting solids, when heated to a temperature in the melting point range, liquefy to act as a binder or granulating medium. The liquefied solid spreads itself over the surface of powdered materials with which it is contacted, and on cooling, forms a solid granulated mass in which the initial materials are bound together. The resulting melt granulation may then be provided to a tablet press or be encapsulated for preparing the oral dosage form. Melt granulation improves the dissolution rate and bioavailability of an active (i.e. drug) by forming a solid dispersion or solid solution.

U.S. Patent No. 5,169,645 discloses directly compressible wax-containing granules having improved flow properties. The granules are obtained when waxes are admixed in the melt with certain flow improving additives, followed by cooling and granulation of the admixture. In certain embodiments, only the wax itself melts in the melt combination of the wax(es) and additives(s), and in other cases both the wax(es) and the additives(s) will melt.

The present invention also includes a multi-layer tablet comprising a layer providing for the delayed release of one or more compounds of the invention, and a further layer providing for the immediate release of a medication for treatment of a disease or disorder. Using a wax/pH-sensitive polymer mix, a gastric insoluble composition may be obtained in which the active ingredient is entrapped, ensuring its delayed release. Parenteral Administration

For parenteral administration, the compounds may be formulated for injection or infusion, for example, intravenous, intramuscular or subcutaneous injection or infusion, or for administration in a bolus dose and/or continuous infusion. Solutions, suspensions or emulsions in an oily or aqueous vehicle, optionally containing other formulatory agents such as suspending, stabilizing and/or dispersing agents may be used. Additional Administration Forms

Additional dosage forms of this invention include dosage forms as described in U.S. Patents Nos. 6,340,475, 6,488,962, 6,451,808, 5,972,389, 5,582,837, and 5,007,790. Additional dosage forms of this invention also include dosage forms as described in U.S. Patent Applications Nos.2003/0147952, 2003/0104062, 2003/0104053, 2003/0044466, 2003/0039688, and 2002/0051820. Additional dosage forms of this invention also include dosage forms as described in PCT Applications Nos. WO 03/35041, WO 03/35040, WO 03/35029, WO 03/35177, WO 03/35039, WO 02/96404, WO 02/32416, WO 01/97783, WO 01/56544, WO 01/32217, WO 98/55107, WO 98/11879, WO 97/47285, WO 93/18755, and WO 90/11757. Controlled Release Formulations and Drug Delivery Systems

In certain embodiments, the formulations of the present invention may be, but are not limited to, short-term, rapid-offset, as well as controlled, for example, sustained release, delayed release and pulsatile release formulations.

The term sustained release is used in its conventional sense to refer to a drug formulation that provides for gradual release of a drug over an extended period of time, and that may, although not necessarily, result in substantially constant blood levels of a drug over an extended time period. The period of time may be as long as a month or more and should be a release which is longer that the same amount of agent administered in bolus form. For sustained release, the compounds may be formulated with a suitable polymer or hydrophobic material which provides sustained release properties to the compounds. As such, the compounds for use the method of the invention may be

administered in the form of microparticles, for example, by injection or in the form of wafers or discs by implantation.

In certain embodiments of the invention, the compounds of the invention are administered to a patient, alone or in combination with another pharmaceutical agent, using a sustained release formulation.

The term delayed release is used herein in its conventional sense to refer to a drug formulation that provides for an initial release of the drug after some delay following drug administration and that may, although not necessarily, includes a delay of from about 10 minutes up to about 12 hours.

The term pulsatile release is used herein in its conventional sense to refer to a drug formulation that provides release of the drug in such a way as to produce pulsed plasma profiles of the drug after drug administration.

The term immediate release is used in its conventional sense to refer to a drug formulation that provides for release of the drug immediately after drug administration.

As used herein, short-term refers to any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes and any or all whole or partial increments thereof after drug administration after drug

administration.

As used herein, rapid-offset refers to any period of time up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes, and any and all whole or partial increments thereof after drug administration. Dosing

The therapeutically effective amount or dose of a compound will depend on the age, sex and weight of the patient, the current medical condition of the patient and the progression of the disease or disorder in the patient being treated. The skilled artisan will be able to determine appropriate dosages depending on these and other factors.

A suitable dose of a compound of the present invention may be in the range of from about 0.01 mg to about 5,000 mg per day, such as from about 0.1 mg to about 1 ,000 mg, for example, from about 1 mg to about 500 mg, such as about 5 mg to about 250 mg per day. The dose may be administered in a single dosage or in multiple dosages, for example from 1 to 4 or more times per day. When multiple dosages are used, the amount of each dosage may be the same or different. For example, a dose of 1 mg per day may be administered as two 0.5 mg doses, with about a 12-hour interval between doses.

It is understood that the amount of compound dosed per day may be administered, in non-limiting examples, every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days. For example, with every other day administration, a 5 mg per day dose may be initiated on Monday with a first subsequent 5 mg per day dose administered on Wednesday, a second subsequent 5 mg per day dose administered on Friday, and so on.

The compounds for use in the method of the invention may be formulated in unit dosage form. The term“unit dosage form” refers to physically discrete units suitable as unitary dosage for patients undergoing treatment, with each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, optionally in association with a suitable pharmaceutical carrier. The unit dosage form may be for a single daily dose or one of multiple daily doses (e.g., about 1 to 4 or more times per day). When multiple daily doses are used, the unit dosage form may be the same or different for each dose.

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents were considered to be within the scope of this invention and covered by the claims appended hereto. For example, it should be understood, that modifications in reaction conditions, including but not limited to reaction times, reaction size/volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g., nitrogen atmosphere, and reducing/oxidizing agents, with art- recognized alternatives and using no more than routine experimentation, are within the scope of the present application.

It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present invention. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

The following examples further illustrate aspects of the present invention. However, they are in no way a limitation of the teachings or disclosure of the present invention as set forth herein.

EXAMPLES

The invention is now described with reference to the following Examples. These Examples are provided for the purpose of illustration only, and the invention is not limited to these Examples, but rather encompasses all variations that are evident as a result of the teachings provided herein. Materials and Methods: Subjects

A total of 268 adult male and female littermate mice were used for all experiments. Mice were maintained under a 12-hour light/dark cycle with ad libitum access to food and water. For behavioral experiments and electrophysiological studies, C57Bl/ 6:129S/J F1 hybrid mice were obtained from the Jackson Laboratory (Bar Harbor, Maine) or bred in house. For CRFr1 colocalization studies, mice with fluorescent-labeled CRFr1 containing neurons were generated as previously described (Justice, et al., J Comp Neurol 2008, 511 :479–496). Mice received implantation between ages 7 and 8 weeks, were allowed to recover for at least 1 week, and were behaviorally tested in age-matched cohorts at 8–20 weeks of age. Mice were singly housed after cannulation to prevent disturbance of the cannula. For electrophysiological experiments, slices were obtained from mice at 9–13 weeks of age. Mice were individually housed for 7–12 days before recording, to mimic the housing conditions of behavioral studies. All studies were conducted in accordance with experimental protocols approved by the University of Pennsylvania Institutional Animal Use and Care Committee and, where applicable, by the Institutional Animal Care and Use Committee of the Weizmann Institute of Science. Stereotaxic Surgery and Placement Verification

Mice were anesthetized with isofluorane and implanted with a 26-gauge guide cannula (Plastics One, Roanoke, Virginia) with a stereotaxic instrument (Kopf, Tujunga, California) positioned 1 mm from the DR with the following coordinates (from brain surface): AP 4.36 mm, ML 1.5 mm, DV 2.0 mm, angled 26 degrees. At the end of each study, mice were transcardially perfused, and cannula placement was verified on the basis of the termination point of the injector as estimated from the location of scar tissue in 50-μm sections through the DR. Mice with incorrect cannula placement were dropped from the statistical analysis. Group sizes reported represent the final group size after subjects with incorrect placements were omitted. Drugs and Microinfusion

All drugs were reconstituted in distilled water, aliquotted, and frozen until the day of use. Fresh aliquots were dissolved in artificial cerebrospinal fluid (ACSF) (Tocris, Bristol, United Kingdom) immediately before behavioral testing. NBI 35965 (Tocris), a highly selective CRFr1 antagonist, was used at 0.44 ng, 1000 times the Ki. Ovine CRF (Sigma, St. Louis, Missouri) was used because of its higher affinity for CRFr1. Doses (1 ng and 50 ng) were selected on the basis of previous studies of DR infusion of this peptide to preferentially target CRFr1. Drug in 0.25

ACSF was infused over 1 min through a microinjector attached to polyethylene tubing connected to a 10

Hamilton syringe on an infusion pump (KD Scientific, Holliston, Massachusetts). Drug or ACSF (.50 μL) was perfused through the microinjector to ensure patency between injections. Hypothalamic-Pituitary-Adrenal Axis Assessment

Testing was performed during a 4-hour period beginning 1 hour after lights- on. Tail blood (10 μL) was collected immediately before DR infusion and at 30, 45, 60, and 120 min post injection. Between the 30- and 45-min collections, mice in the NBI 35965 study were restrained in a 50-mL conical tube with a 5-mm air hole. Corticosterone was measured as described previously (Gerber, et al., Endocrinology 2012, 153:4830–4837). Behavioral Testing

The tail suspension test (TST) and light-dark box (LD) were performed on separate cohorts of mice 30 min after drug or ACSF infusion. Methods were similar to those described previously (McEuen, et al., J Neurosci 2008, 28:8169–8177; Bale, et al., Nat Genet 2000, 24:410–414). cFos Immunohistochemistry

To assess CRF-induced neuronal activation in the DR, double labeling immunohistochemistry for cFos and tryptophan hydroxylase (TPH) was performed on DR sections from mice sacrificed 90 min after CRF or ACSF infusion. Methods were similar to those described previously (Goel, et al., Endocrinology 152:2001–2010; Faulconbridge, et al., Brain Res 2008, 1218:151–157). Gene Expression Analysis

Brains were collected from experimentally naive adult male and female mice. Female brains were collected in diestrus. Gene expression of CRFr1, CRFr2, CRF binding protein, TPH2, and γ- aminobutyric acid (GABA) receptor subunits alpha-1 , alpha-2, delta, and gamma-2 were determined by quantitative Taqman real-time polymerase chain reaction as previously described (Paxinos, et al., 2004, The Mouse Brain in Stereotaxic Coordinates, Compact 2nd ed. Boston: Elsevier Academic Press; Rodgers, et al., J Neurosci 2013, 33:9003–9012). Immunofluorescence and CRFr1 Localization

Dual immunofluorescence was performed to detect enhanced green fluorescent protein (GFP) and TPH or parvalbumin in DR sections from paraformaldehyde- fixed male and female CRFr1- GFP mice. Electrophysiology

A modified procedure based on the method of Challis et al., Raphe

GABAergic neurons mediate the acquisition of avoidance after social defeat; J Neurosci 2013, 33:13978–13988 (52) was used. Data Analysis and Statistics

Total corticosterone was analyzed by multivariate analysis of variance (ANOVA) (drug x time). Behavioral measures were analyzed by two-way ANOVA (sex x drug). Subsequent one way analyses were performed on data within sex, with Dunnett’s test used to identify significant post hoc comparisons. Student t test was used to compare gene expression between males and females. To determine CRFr1 counts, a generalized linear mixed model was employed to analyze GFP count x sex x subregion with a Poisson distribution. Assuming a binomial distribution, further analyses were made to predict the likelihood that a given GFP-immunoreactive (-ir) cell co-expressed parvalbumin or TPH. Data are reported as estimated effect size ± 95% confidence intervals. Significance was determined as p <0.05, with 95% confidence intervals not bounding zero. Statistics were performed in R software. For electrophysiological studies, results of 5-HTergic neuron response to CRF were compared between males and females via two-way repeated measures ANOVA and post hoc Tukey tests employing sex and drug (6,7-dinitroquinoxaline-2, 3-dione [DNQX] vs. DNQX + CRF) as the independent variables. Statistics were performed with JMP8 (SAS, Cary, North Carolina) software; data are reported as mean ± SEM. Example 1: DR Infusions of NBI 35965 or CRF Preferentially Alter Male

Corticosterone Production

The 5-HT output from the DR has modulatory activity on the hypothalamic- pituitary-adrenal (HPA) axis. The CRF regulation of DR neurons could therefore influence HPA responsiveness. Thus, the effect of CRFr1 antagonism within the DR on the

corticosterone response to restraint stress was assessed (FIG. 1). The NBI significantly blunted corticosterone levels in males (F_1,9 = 7.085, p =0.026). The effect of NBI was manifested as a reduction in the rise time from 0 to 30 min before restraint (t₉ = 3.191, p =0.011) and a reduction in total corticosterone produced throughout the course of the experiment (area under the curve) (t₉ = 2.794, p = 0.021). In females, NBI did not significantly impact corticosterone production (F_1,10 = 0.1180, p = 0.7383). The effect of CRF infusion on the HPA axis was tested next. In males, CRF significantly increased corticosterone (F_1,17 = 5.926, p = 0.026). In females, CRF did not significantly affect corticosterone (F_1,18 = 1.28, p =0.27). Example 2: Male-Specific Effects of DR Infusion of NBI 35965 and CRF on Stress Coping and Anxiety-Like Behaviors

To assess the role of DR CRFr1 in modulating stress coping behavior of male and female mice, the TST was performed 30 min after infusion of NBI 35965 or vehicle (ACSF) (FIG.2). It was detected that a significant interaction of sex and NBI on latency become immobile (F_1,37 = 6.654, p = 0.014), where NBI increased latency in males (p = 0.015) but not in females (p = 0.37). There was a trend toward an interaction effect of NBI on immobility in the TST, where NBI reduced immobility in males and increased immobility in females, although this effect failed to reach statistical significance (F_1,40 = 2.651 , p = 0.11). The observed sex difference in response to the CRFr1 antagonist suggested potential sex differences in the response to CRF. To address this possibility, the effect of two doses (1 ng, 50 ng) of CRF infusion on behavior was assessed in the TST. The 1-ng dose of CRF was ineffective to change immobility or latency to become immobile on either test, suggesting ineffective local concentrations were achieved. The 50-ng dose of CRF decreased immobility (F_2,26 = 3.467, p = 0.046) and increased latency to become immobile (F_2,26 = 8.684, p = 0.001 ) in males and was without effect in females. To assess anxiety-like behavior, mice were tested in the LD. The NBI had no effect in either sex on any parameter. However, as with behavioral outcomes in the TST, where 1-ng dose CRF was without effect, 50-ng dose CRF had sex-specific effects on behavior, increasing latency to exit the light compartment (F_2,26 = 5.313, p = 0.011 ), and total time spent in the light compartment (F_2,26 = 5.024, p = 0.014) in males but not females. In males, 50 ng CRF also reduced the number of transitions between compartments (F_2,26 = 4.915, p = 0.016) but did not affect distance traveled in the light (normalized to time spent in the light), indicating that the animals did not freeze while in the light compartment. Example 3: DR Infusion of CRF Increases cFos in Males But Decreases cFos in

Females

To determine whether sex differences in behavioral responsiveness to CRF in the DR were associated with differential patterns of neuronal activation, cFos

immunoreactivity after infusion of CRF or ACSF was assessed (FIG. 3, Table 1). In males, CRF increased the number of cFos-positive cells across the DR (F_1,65 = 14.79, p < 0.001), whereas in females, CRF reduced the number of cFos-positive cells (F_1,56 = 7.563, p = 0.008). Subregion analysis indicated that this interaction was present in dorsomedial (F1,17 = 6.216, p = 0.023), ventromedial (F_1,19 = 5.590, p = 0.029), and lateral wing subregions of the DR (rostral, F_1,24 = 7.300, p = 0.013; caudal, F_1,21 = 7.359, p = 0.013). The CRF had a similar effect of increasing cFos-positive cells in both male and female rostral DR (F_1,21 = 12.38, p = 0.002), and no significant main effects were found in the caudal DR

 Example 4: CRFr1 Is Expressed Differentially Throughout Subdivisions of the DR in Males and Females

An experiment was conducted to determine whether the observed sex differences in responsiveness to the CRFr1 antagonist or CRF was due to differences in transcript levels of genes relevant to CRF and 5-HT signaling. The relative expression of CRFr1 , CRFr2, and CRF- binding protein messenger RNA (mRNA) was quantified in DR micropunches from

experimentally naïve male and female mice (FIG. 4, Panel A). No difference was observed in the mRNA levels of any of these relevant transcripts. Because it was predicted that some of the observed behavioral and physiological differences might be GABA-mediated, the relative expression of GABA receptor subunits alpha-1, alpha-2, delta, and gamma-2 was also quantified, which play important roles in receptor kinetics. No sex difference was observed in mRNA in any of these receptor subunits. Differences in TPH2 was detected, with females expressing 1.62- fold higher levels relative to males (t_1,8 = 2.652, p = 0.029).

After finding no differences in CRFr1 gene expression, it was hypothesized that sex differences in responsivity to CRFr1 antagonist or CRF might be due to differences in the neurotransmitter cell type expressing CRFr1. Because current available antibodies are unable to distinguish between CRFr1 and CRFr2, a CRFr1-GFP transgenic mouse was used in which GFP is transcribed under the control of the CRFr1 promoter to identify CRFr1 positive neurons in the DR (FIG. 4, Panels B-N). Sections throughout the DR from these mice were dual labeled for either GFP and parvalbumin to identify putative GABAergic neurons expressing CRFr1 or GFP and TPH to identify serotonergic neurons expressing CRFr1. In accordance with the CRFr1 mRNA data, there were no sex differences in overall number of GFP-ir cells (-0.542 ± -1.59, +0.053; p = 0.0796). However, a sex difference was observed in co-localization of GFP-ir cells throughout regions of the DR. In females, the probability that a given GFP-ir cell coexpressed parvalbumin was lower than in males (-2.535 ± 4.819, -0.259; p = 0.0291). The probability that a given GFP-ir cell co-expressed TPH was < 25% in all subdivisions, regardless of sex, except within the rDR, where males displayed significantly more co-localization with TPH than females (-2.16 ± 3.736, -0.583; p = 0.007). Example 5: 5-HT Neurons in Females Demonstrate Reduced Excitability and a Blunted Response to CRF

To investigate physiological differences in DR 5-HT neurons of males and females, whole-cell electrophysiological recordings were conducted with current-clamp and voltage-clamp techniques (FIG. 5 , Table 2). Data from a total of 21 neurons, with current-clamp recordings, were analyzed (9 from 6 male mice, 12 from 7 female mice). Cellular characteristics that were measured, included resting membrane potential, resistance, time constant (tau), after hyperpolarization, and action potential amplitude width, and threshold. Characterization of male and female 5-HT neurons revealed females to have a significantly depolarized resting membrane potential (t₁₃ = 2.498, p = 0.027) and a reduced tau (t₁₂ = 2.225, p = 0.046) relative to males. No other membrane characteristics differed between males and females. However, a nonlinear regression on the frequency-current (F-I) plot of male and female 5-HT neurons revealed male neurons to have a greater excitability (slope) compared with females (F_1,158 = 8.929, p = 0.003), firing at 27.28 Hz versus 14.83 Hz, to a maximal injected current of 160 pA. Voltage-clamp recordings of GABAergic inhibitory postsynaptic currents (IPSCs), isolated by the addition of DNQX, revealed several functional differences between male and female 5-HT neurons. In response to the addition of CRF (30 nmol/L), males and females showed differential changes in the initial first decay time (t₁₀ = 2.673, p = 0.023), with males showing an increase (+2.148 ± 1.376) and females showing a decrease (-2.380 ± 1.045) compared with DNQX alone. Males and females also exhibited a divergent CRF mediated change in rise time (t₁₁ = 2.802, p = 0.0187), and half width (t₁₁ = 5.994, p <0.0001); in both measures males showed an increase, whereas females showed a decrease.

Results

Male and female mice received an infusion of the CRFrl small molecule antagonist, NBI 35965, or one of two doses of CRF directly into the DR and were evaluated for changes in physiological and behavioral stress responsiveness. The NBI 35965 in the DR significantly blunted corticosterone levels in response to a restraint stress only in males.

Similarly, CRF infused into the DR in the absence of restraint significantly elevated

corticosterone production above the levels induced by intracranial injection only in males. The 5-HT system is a known activator of the HPA axis, where selective SSRIs and 5-HT agonists increase corticosterone production during and independent of stress. Although direct innervation of the paraventricular nucleus (PVN) has been reported (Petrov, et al, Cell Tissue Res 1994,

277:289-295; Petrov, et al, J Comp Neurol 1992, 318: 18-26; Williamson, et al, J Comp Neurol 2007, 503:717-740), 5-HTergic fibers from the DR also heavily innervate the GABAergic neurons of the PVNsurround (Sawchenko, et al, Brain Res 1983, 277:355-36064). Therefore, CRF -mediated changes in 5-HT output could modulate this axis through a disinhibition of medial parvocellular neurons. Thus, a male-specific response to administration of NBI 35965 and CRF directly into the DR suggests a unique sex-specific mechanism upstream of the PVN, including potential differences in CRFrl co-localization or signaling within the DR

In the assessment of behavioral stress coping strategies, including the TST, only males again showed a significant effect of NBI 35965 to increase the latency to immobility and of CRF infusion to reduce the immobile time. Although NBI 35965 infusion produced no significant changes in male or female mice in the LD, CRF at the higher dose again increased time spent in the light and escape latency only in males. These findings are consistent with behavioral effects reported in previous studies for systemically administered CRFrl antagonists and CRFrl gene deletion, implicating the DR as a key brain region mediating these outcomes. Interestingly, in current studies, both CRF and NBI 35965 infusions produced similar effects in the TST.

42 These sex differences were most apparent in the dorsomedial and lateral wings of the DR, regions previously implicated in uncontrollable stress and anxiety.

Immunohistochemical evidence supports a dense CRF innervation of these DR subregions, where CRF fibers primarily contact GABA-containing dendrites. Thus, differences in the number of CRF responsive GABAergic neurons in females could account for the observed reduction in activation detected in these mice. Within the DR, there is a well-described topographical organization, where GABAergic interneurons exhibit tight control over the tonic and activity-mediated release of 5-HT. To test the hypothesis that divergent sex responses to CRF application within the DR might be due to differences in CRFr1 localization on functionally distinct neuronal populations, a CRFr1 - driven GFP transgenic reporter mouse line was used to quantify sex differences in co-expression of CRFr1. Because the available antibodies for the CRF receptors are known to be of poor quality and lack sufficient specificity, this reporter mouse provided an excellent tool to identify CRFr1 -positive neurons in the DR. Sex differences was quantified in co-expression of GFP with TPH, a marker of 5-HTergic neurons, or with parvalbumin, a marker of a subset of GABAergic neurons. Parvalbumin was used to mark GABAergic neurons, because 87% of GAD67-ir neurons in the DR co-express parvalbumin. These neurons might display some functional differences compared with the broader population of GABAergic neurons but were selected on the basis of reliable somal immunoreactivity for co- expression analysis with CRFr1. Overall numbers of CRFr1 neurons did not differ between males and females, as indicated by similar numbers of GFPpositive neurons in the DR. This was also confirmed by similar expression levels of CRFr1 mRNA in the DR of males and females. As expected, few GFP-positive neurons were co-expressed with TPH, consistent with previous reports that demonstrate CRF primarily acts on GABAergic neurons and that CRFr1 shows little expression overlap with 5-HT neurons. Surprisingly, females had reduced GFP co-labeling with parvalbumin compared with males. This outcome supports a revised model; rather than females demonstrating greater CRFr1-mediated GABA tone, sex differences in CRF-induced neuronal activation might be due to differences in CRFr1 intracellular signaling, trafficking, or receptor kinetics. Alternatively, CRF might be activating a population of parvalbumin-negative GABA neurons in the female DR.

On the basis of the observed sex differences in CRFr1 coexpression, without being bound to any specific theory, it was hypothesized that DR 5-HT neurons in males and females might receive differential GABA input in response to CRF. To examine this, whole-cell electrophysiological recordings was used to measure GABAergic IPSCs in 5-HT neurons before and after application of CRF. A striking sex difference was observed in CRF responsivity, where CRF increased IPSC decay time in males but decreased decay time in females. The IPSC decay time can correlate with the number of activated axonal inputs during the IPSC, suggesting sex differences in presynaptic GABAergic input onto 5-HT neurons. This is consistent with the differential coexpression of CRFrl on parvalbumin neurons between males and females.

Furthermore, the reduced IPSC half-width in response to CRF exhibited by females also supports this sex difference in the number of GABAergic release sites onto 5-HT neurons. These differences in IPSC kinetics constitute a significant functional difference between males and females that could alter 5-HT neuron excitability and neurotransmission and thereby influence stress physiological and behavioral measures.

In gathering basal characteristics before assessing sex differences in 5-HTergic neuron responses to CRF, an unexpected sex difference was discovered in 5-HT neuronal excitability. With whole-cell patch clamp recordings from 5-HT neurons from male and female dorsomedial DR slices, reduced neuronal excitability was found in females in response to a series of current injections. In addition to sex differences in CRFrl localization and GABAergic responses to CRF, this suggests that 5-HT neurons in females require a greater depolarizing stimulus to generate neuronal firing and subsequent 5-HT release, even at baseline. Compared with males, this might translate to 5-HTergic hypofunction in females, an underlying risk factor for the development of affective disorders during stress exposure.

Overall, the results of these experiments reveals intriguing sex -based differences in behavioral and physiological effects of a CRFrl antagonist and CRF in the DR, which were mechanistically associated with sex differences in receptor co-expression and divergent electrophysiological responses to CRF in 5-HTergic neurons. The blunted response of females points to a potential explanation for the lack of efficacy in CRFrl antagonists in clinical trials, which have focused primarily on female participants due to their increased disease prevalence (Table 3). These findings support the importance of identifying sex differences in central stress pathways to understand the heightened predisposition of females toward these disorders and in identifying sex appropriate and potentially sex-specific pharmacological treatments.

The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety.

While the invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

CLAIMS What is claimed is:

1. A method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist, or a salt or solvate thereof, wherein the disorder manifests hypersecretion of corticotropin- releasing factor (CRF) and the disorder is selected from the group consisting of an anxiety-related disorder, a mood disorder, a post-traumatic stress disorder, immune suppression, drug or alcohol withdrawal symptoms, a sleep disorder induced by stress, fibromyalgia, dysthemia, a bipolar disorder, cyclothymia, fatigue syndrome, stress-induced irritable bowel syndrome, and stress-induced headache.

2. The method of claim 1, wherein the disorder is selected from the group consisting of an anxiety-related disorder, a mood disorder, a bipolar disorder, and a post- traumatic stress disorder.

3. The method of claim 2, wherein the anxiety-related disorder is selected from the group consisting of an anxiety state, a generalized anxiety disorder, a social anxiety disorder, an anxiety with co-morbid depressive illness, a panic disorder, an obsessive-compulsive disorder, a phobic disorder, a post-traumatic stress disorder, and an atypical anxiety disorder.

4. The method of claim 2, wherein the mood disorder is depression selected from the group consisting of major depression, single episode depression, recurrent depression, child abuse induced depression, and postpartum depression.

5. The method of claim 1, wherein the CRFr1 antagonist is selected from the group consisting of LY2371712, NBI-35965, NBI-30775, Antalarmin, CP-316,311, CRA 5626, CP-154,526, Emicerfont, ONO-2333Ms, Pexacerfont, SSR125543, NBI-34101 , DMP-696, DMP-904, DMP-695, SC-241, BMS-561388, R121919, NBI-30545, CP-376,396, NBI-27914, NBI-34101 , PF-572778, GSK561579, and GSK586529.

6. A method of treating a disorder in a male subject, comprising administering to the male subject a pharmaceutically effective amount of a CRFr1 antagonist, or a salt or solvate thereof, and a second therapeutic agent or a salt or solvate thereof, wherein the disorder manifests hypersecretion of corticotropin-releasing factor (CRF) and the second therapeutic agent is selected from the group consisting of sertraline, fluoxetine, citalopram, escitalopram, paroxetine, fluvoxamine, trazodone, desvenlafaxine , duloxetine, venlafaxine, amitriptyline, amoxapine, clomipramine, desipramine, doxepin, imipramine, nortriptyline, protriptyline, trimipramine, maprotiline, nefazodone, trazodone, isocarboxazid, phenelzine, selegiline, tranylcypromine, mirtazapine, quetiapene, buspirone, bupropion, a selective serotonin reuptake inhibitor, a serotonin and norepinephrine reuptake inhibitor, a tricyclic antidepressant, a tetracyclic antidepressant, a dopamine reuptake blocker, a 5-HT2 receptor antagonist, a 5-HT3 receptor antagonist, a monoamine oxidase inhibitor, a noradrenergic antagonist, and a mixture thereof.

7. The method of claim 1, wherein the male subject is a male human.

8. The method of claim 7, wherein the CRFr1 antagonist is simultaneously

administered with the second therapeutic agent.

9. The method of claim 6, wherein the CRFr1 antagonist is administered 1-6 hours before the administration of the second therapeutic agent.

10. The method of claim 6, wherein the CRFr1 antagonist is administered 1-6 hours after the administration of the second therapeutic agent.