METHODS FOR SCREENING GABA-MODULATORY COMPOUNDS FOR SPECIFIED PHARMACOLOGICAL ACTIVITIES

Abstract: Methods are provided that are useful in assaying compounds for cognitive enhancing properties, anxiolytic properties, hypnotic properties, or antidepressant properties. These methods involve determining the <i>in vitro</i> efficacy and EC50 of the compounds at defined series of cloned GABAA subtype receptors composed of specific variants of alpha , beta , and gamma subunits in order to develop and an activity profile for each compound. Optionally, the binding affinities of the compounds at GABAA receptors are also determined. As an additional step the <i>in vivo</i> effects of the compounds may be tested in animal models.

Description: Methods for Screening GABA-Modulatorv Compounds for Specified Pharmacological Activities Cross Reference to Related Applications This application claims priority from U.S. provisional applications 60/133,195, entitled "Methods for Screening Compounds for Cognition Enhancing Activity", 60/133.191, entitled "Methods for Screening Compounds for Antidepressant Activity", 60/133,155, entitled "Methods for Screening Compounds for Hypnotic Activity", and 60/133,154, entitled "Methods for Screening Compounds for Anxiolytic Activity" , each of which was filed m the names of the present inventors on May 7, 1999. To the extent that they differ from the disclosure of the present application, the disclosures (including the claims) of these provisional applications are incorporated herein by reference. BACKGROUND OF THE INVENTION Field of the invention The present invention relates to methods for assaying GABA- modulatory compounds for activity as antidepressants , cognitive enhancers, sedative hypnotics, or non-sedatmg anxiolytics. In particular, the method includes determining efficacy (generally in vitro ef ficacy) and EC 5C values (as used herein incorporating IC 50 values) for the compounds at several different cloned (i.e., expressed m cells as directed by heterologouε cloned receptor- encoding nucleic acid expression vector molecules) GABA subtype receptors (each subtype maαe up cf a defined set of specific receptor subunit isotypes) The method optionally includes determining binding affinity of compounds for GABA A receptors. The method results m the development of an activity profile for each compound. As an additional step, animal models predictive of such effects may be used to measure the ability of compounds to effect cognitive enhancement, to act as antidepressants , to mediate sedative hypnotic effects, or to effect anxiolysis m vivo without eliciting certain undesirable side effects. Description of related art Modern drug discovery methodology allows the testing of large numbers of compounds (often assembled into collections termed libraries) for functional characteristics that confer pharmaceutical utility. This "screening" of such libraries, using specific tests (assays) for functional activity properties, allows the rapid identification of promising compounds for further development as pharmaceutical agents. There has been a longstanding quest m the pharmaceutical industry for new means of identifying such promising compounds. Such new means may involve new assays, or may use old assays to generate data that can be analyzed and applied m new ways to identify compounds with new and useful characteristics. In the field cf psychopharmacology, the use of cloned neuronal receptors as substrates has provided new, more specific assays with which compounds can oe characterized The use of such receptors has enabled the development of receptor binding profile criteria that are particularly beneficial m the identification of useful psychopharmacological agents. For example, such profiling can identify compounds that will be free of certain undesirable adverse effects (side effects) . The GABA. receptor superfamily represents one of the classes of receptors through which the major inhibitory neurotransmitter, γ-aminobutyπc acid, or GABA acts. In addition to being the site of neurotransmitter action a number of drugs including the anxiolytic and sedating benzodiazepin.es bind to this receptor. The GABA A receptor is a chloride channel that opens m response to GABA, allowing chloride to enter the cell. This effects a slowing of neuronal activity through hyperpolarization of the cell membrane potential. GABA A receptors are composed of several protein subunits and are generally pentameric in structure. A number of cDNAs for GABA A receptor subunits have been cloned. While these subunits share a basic motif of 4 membrane- spanning helices, there is sufficient sequence diversity to classify them into several groups. To date at least 6α, 3β, 3γ, lε, lδ, 2p, and 1~ subunit species have been identified; some representing alternatively spliced forms. Native GABA. receptors are typically composed of α, β, and γ subunits, most often m the ratio of two alphas, two betas, and one gamma, although other combinations (some comprising other subunits such as ε, δ, p, or π) have been described. Even if restricted to only α, β, and γ subunits, however, an enormous diversity of GABA A subtype receptors are possible. Evidence such as message distribution, genome localization and biochemical studies suggests that the major naturally occurring receptor combinations are α,β 2 γ 2 , α : β 3 γ 2 , α 3 β 3 γ 2 , and α 5 β 3 γ 2 . In the typical GABA A receptor, the binding sites for GABA (2 per receptor complex) are formed by ammo acids from the α and β subunits. Amino acids from the α and γ subunits contribute to form 1 benzodiazepine site per receptor complex. In a classic allosteric mechanism, the binding of a drug to the benzodiazepine site increases the affinity of GABA binding to the receptor. Benzodiazepines and related drugs that enhance the ability of GABA to open GABA A receptor channels are known as agonists or partial agonists depending on the level of enhancement. Other classes of drugs such as β-carboline derivatives that occupy the same site and negatively modulate the action of GABA are called inverse agonists. A third class of compounds exists that occupies the same site as both the agonists and inverse agonists (blocking access of these agents to the site) and yet has little or no direct effect on GABA activity. These compounds are referred to as antagonists. The characterization of activities of different subtype receptors has been an area of intense pharmacological discovery. Agonists that act at the benzodiazepine site have long been known to know to exhibit anxiolytic, sedative, and hypnotic effects animal behavior models, while compounds that act as inverse agonists at this site elicit anxiogemc, cognition enhancing, and proconvulsant effects While benzodiazep es have long been used as anxiolytics, these compounds exhibit a number of undesirable side effects. These include cognitive impairment, sedation, ataxia, potentiation of ethanol effects, and a tendency for tolerance and drug dependence . Likewise the development of benzodiazepine site ligands for other indications has been thwarted by unfavorable side effect profiles for each indication. For example, compounds known to possess cognition enhancing properties have generally tended to be anxiogemc and proconvulsant, while compounds that produce anxiolytic effects tend to generate unwanted sedation, and do so more powerfully when taken conjunction with the consumption of alcoholic beverages . SUMMARY OF THE INVENTION The present invention provides methods for characterizing compounds that act at the GABA ^ receptor benzodiazepine site. In particular, it provides methods for identifying compounds with characteristics indicating that the identified compounds will exhibit pharmacological properties consistent with their use as antidepressants , cognitive enhancers without anxiogemc or proconvulsant activity, sedative hypnotics without cognition- lmpairmg activity, or non- sedating anxiolytics. This invention is useful screening libraries of compounds for therapeutic potential and m drug design efforts. Measurement of GABA receptor binding affinity is a useful step any of the methods of the invention. Alternatively, these assays may be performed without measuring the binding affinity of the compound. The assays may include an assessment of the ability of the compound to mediate the desired effects m vivo without eliciting side effects using animal models established to be predictive of the desired effects and animal models predictive of the undesired side effects that have been associated with other compounds acting at GABA. receptors. As used herein, the term "efficacy" refers to amount of potentiation (represented as a % increase, e.g., 10%) or inhibition (represented as a % decrease, e.g., -10%) of GABA activated responses measured for GABA A receptors . In addition to the ability of a compound to effect a specified magnitude of change m the GABA response at distinct subtype receptors, the EC„ value of the compound at the pertinent receptors is also taken into consideration. As used herein, the term "EC 50 " or "EC ^ value" refers to the concentration of a compound needed to elicit half the maximal response (to the agonist or inverse agonist effects of a compound) that can be obtained with the compound. Thus, a compound that exhibits dissimilar EC^ values at different subtype receptors can selectively potentiate one of those receptors over a defined range of drug concentrations, even though the maximal amount of potentiation achievable by the compound is the same for the two subtype receptors over a much broader range of compound concentrations. EC ^0 values do not necessarily correlate to binding affinities or to compound efficacies. Cognitive Enhancers: With regard to identifying cognitive enhancers, a method of the invention involves optionally determining the binding affinity of a compound for GABA A receptors having Rol5-1788 binding sites and determining efficacy and EC cr values for the compound at cloned a^ 2 y 2 and α 5 β 3 γ 2 receptors and comparing these values with efficacy and EC 50 values for the compound at cloned GABA A receptors containing the α 2 or α 3 subunits. The ability of the compound to mediate cognitive enhancing effects may optionally be assessed m vivo by animal model predictive of cognitive enhancement. Whether the compound causes proconvulsant effects may also be assessed m vivo using animal models for detecting proconvulsant activity. Accordingly, one aspect this invention provides methods for identifying compounds with cognitive enhancing activity that do not display the side effects of anxiogenesiε or proconvulsant activity. These methods comprise: a) Screening compounds, optionally determining the binding affinity of the compounds for GABA A receptors; b) determining in vi tro efficacy and EC 50 values of the compounds using cloned α^y ^ and α 5 β 3 y 2 receptors and comparing these values to m vi tro efficacy and EC 50 values for the compound determined using GABA A receptors that contain the α, or α subunit; and c) selecting compounds having significant inverse agonist character and sufficiently low EC 5C values at α ^ β 2 γ 2 or α 5 β 3 γ 2 subtype receptors and that produce agonist activity at GABA A subtype receptors that contain the α 2 or ct, subunit . Thus, the invention presents novel methods for identifying compounds with selective cognitive enhancing properties (1) by examining the Dinding of a given compound at GABA_ receptors and (2) by assessing the ability of the compound to potentiate GABA responses at a series of GABA A subtype receptors. These values are then compared to a set of criteria termed the "Cognitive Enhancer Activity Range Profiler" (Table I, below). This activity profile comprises measurements of m vitro efficacy (agonist, inverse agonist or antagonist character) and EC 50 values at each of 4 GABA A subtype receptors . The activity profile needed for cognitive enhancement is presented as a precise window of inverse agonism at certain subtype receptors and agonism at other subtype receptors. The EC 50 criteria at each of these subtype receptors are also presented. Determining efficacy and EC 50 values for a test compound is crucial as many compounds bind with high affinity at the benzodiazepine site without potentiating the GABA response at the appropriate subtype receptors. In certain embodiments, the conclusions drawn from the in vi tro determinations of validity may be confirmed by examining the m vivo efficacy of test compounds as cognitive enhancers using animal models for cognitive enhancement. It may also be necessary to verify that compounds identified by these methods possess the predicted favorable side effect profiles by examining the performance of the compounds in animal models indicative of these side effects. Thus, the animal models may be used as an additional step of the assay to further refine the selection of compounds with cognitive enhancing activity. Anxiolytics : With regard to identifying anxiolytics, a method of the invention involves optionally measuring the binding affinity of a compound at GABA A receptors having Rol5-1788 binding sites and measuring the efficacy and EC 50 values for a compound using cloned o β 3 γ ^ and ,β 3 y ^ receptors and comparing these values with the activities and EC,, values of the compound at cloned GABA A receptors containing the α 2 or α r subunits. As an additional step, the ability of the compound to mediate anxiolytic effects may be assessed m vivo using an animal model established to be predictive of anxiety, and whether the compound causes sedative effects may also be assessed m vivo by an animal model shown to measure sedation. Accordingly, this invention provides a method for identifying compounds with anxiolytic activity that do not display the side effects of cognitive impairment, ataxia, potentiation of alcohol effects, and a tendency for tolerance and drug dependence or that display these side effects only to a very minimal degree. These method comprise: a) screening compounds, optionally measuring the binding affinity of the compounds at GABA A receptors; b) measuring the m vi tro efficacy and EC c values of the compounds at cloned α 2 β 3 y <; and α,β 3 t receptors and comparing these values to the m vi tro efficacy and EC k „ values of the compounds at GABA A receptors that contain the α or α 5 subunit, and c) selecting compounds with partial agonist character and sufficiently low EC CC values at 2 β 3 γ ^ and α 3 β 3 γ 2 subtype receptors that also display lower activity at GABA A subtype receptors that contain the α or α subunit Alternatively, this assay may be performed without measuring the binding affinity of the compound or with the additional step of assessing the ability of the compound to mediate anxiolytic effects m vivo without causing sedation via an animal model established to be predictive of anxiety and an animal model predictive of sedative effects . Thus, the invention presents a method for identifying compounds with selective anxiolytic activity (1) by examining the binding of a given compound at GABA A receptors and (2) by assessing the ability of the compound to potentiate GABA responses at a series of GABA A subtype receptors The resulting values are then compared to a set of empirically defined criteria termed the "Anxiolytic Activity Range Profiler" (Table III, below) The criteria given by the Anxiolytic Activity Range Profiler are used to select compounds with anxiolytic activity that have no or very minimal sedative effects. This activity profile comprises determinations of m vi tro efficacy (agonist, inverse agonist or antagonist character) and EC 5C values at each of 4 GABA A subtype receptors. The activity profile needed for anxiolysis is presented as a precise window of agonism and EC 50 criteria at each of these subtype receptors. The combination of determining efficacy, and EC 50 values for a test compound is crucial as many compounds bind with high affinity at the benzodiazepine site without potentiating the GABA response at the appropriate subtype receptors. In certain embodiments, the conclusions drawn from the m vi tro determinations may be confirmed by examining the m vivo efficacy of test compounds predicted to have anxiolytic activity in an animal model for anxiety. It may also be desirable to verify that compounds identified by these methods possess the predicted favorable side effect profiles by examining the performance of the compounds m animal models known to be indicative of sedative effects. Thus, the animal models may be used as an additional step of the assay to further refine the selection of compounds with non-sedating anxiolytic properties. Antidepressants : With regard to identifying antidepressants, a method of the invention involves optionally measuring the binding affinity of a compound at GABA A receptors, measuring the efficacy and EC 5 „ values for a compound at cloned α 2 β 3 γ 2 and 3 β 3 γ 2 receptors, and comparing these values with efficacy and EC 50 values for the compound at cloned GABA A receptors containing the α x or α c subunits. Optionally, the ability of the compound to mediate antidepressant effects may be assessed m vivo using one or more animal models predictive of antidepressant activity. Whether a compound produces sedative effects may also be assessed m vivo with an animal model for measuring sedation. Accordingly a broad aspect, this invention provides a method for screening compounds for antidepressant activity that do not cause the side effects of cognitive impairment, ataxia, potentiation of alcohol effects, and a tendency for tolerance and drug dependence, or that display these side effects only at a very low level. These method comprise: a) screening compounds, optionally measuring the binding affinity of the compounds at GABA A receptors,- b) determining in vi tro efficacy and EC K0 values for the compounds at cloned α 2 β 3 γ 2 and α 3 β 3 γ 2 receptors and comparing these values to m vi tro efficacy and EC 50 values for the compounds determined at GABA A receptors that contain an α x or α 5 subunit ; and c) selecting compounds having partial agonist character and that produce sufficiently low EC 50 values at α 2 β 2 and α 3 β 3 γ 2 subtype receptors and display lower efficacy activity at GABA A subtype receptors that contain the α x or α 5 subunit . The compounds selected by this method have antidepressant activity. The assay may include an assessment of the ability of the compound to mediate antidepressant effects m vivo without causing sedation. Animal models predictive of antidepressant effects and sedative effects can be used for the in vivo determination . Thus, the invention presents methods for identifying compounds with antidepressant activity, i.e., selective antidepressant activity, (1) by examining the binding of a given compound at GABA A receptors and (2) by assessing the ability of the compound to potentiate GABA responses at a series of GABA A subtype receptors. The resulting values are then compared to a set of criteria termed the "Antidepressant Activity Range Profiler" (Table IV, below) . The criteria given by the Antidepressant Activity Rang Profiler are used to identify compounds with antidepressant activity that have no or very minimal sedative effects. This activity profile comprises determinations of in vi tro efficacy (agonist, inverse agonist or antagonist character) and EC 50 values at each of 4 GABA A subtype receptors. The activity profile needed for antidepressant activity is presented as a precise window of agonism and EC S0 values at each of these subtype receptors. The combination of determining efficacy and EC S0 values for a test compound is crucial as many compounds bind with high affinity at the benzodiazepine site without potentiating the GABA response at the appropriate subtype receptors. Furthermore, as noted previously, EC t0 values do not necessarily correlate with binding affinities or compound efficacies. In certain embodiments, the conclusions drawn from the m vi tro determinations may be confirmed by examining the in vivo effects of test compounds selected as having antidepressant activity using animal models for depression. It may also be desirable to verify that compounds identified by these methods do indeed possess the predicted favorable side effect profiles by examining the performance of the compounds animal models known to be indicative of these side effects. Thus, the animal models may be used as an additional step of the assay to further refine the selection of compounds with selective antidepressant activity. Hypnotics : With regard to hypnotics, a method of the invention involves optionally measuring the binding affinity of a compound at GABA A receptors having Rol5-1788 binding sites, and measuring the efficacy and EC, values of a compound at cloned α 2 β 3 γ 2 and α 3 β 3 γ 2 receptors and comparing these values with the activities - and EC εo values of the compound at cloned GABA A receptors containing the α ^ or . subunits. Optionally, the ability of the compound to mediate hypnotic effects is assessed in vivo using an animal model established to be predictive of sedation, and whether the compound causes cognitive impairment may also be assessed in vivo by an animal model shown to be predictive of this effect. Accordingly, this invention provides a method for identifying compounds w th hypnotic activity that do not display the side effects of cognitive impairment, ataxia, potentiation of alcohol effects, and a tendency for tolerance and drug dependence or that display these side effects only at a very low level. This method comprises: a) screening compounds, optionally measuring the binding affinity of the compounds at GABA A receptors; b) measuring the EC 50 and m vi tro efficacy values of the compounds at cloned α 2 β 3 γ ^ and α 3 β 3 y t receptors and comparing these values to the EC, 0 and in vi tro efficacy values of the compounds that contain the α_ or α 5 subunit; and c) selecting compounds with sufficiently low EC 50 values at α 2 β 3Ϊ2 an α 3 β 3 γ 2 receptors, partial agonist activity at α 2 β 3 γ 2 receptors and stronger partial agonist activity at ct 3 β 3 γ 2 receptors, that also display lower activity at GABA A subtype receptors that contain the α or o subunit as having hypnotic activity. Alternatively, this assay may be performed without measuring the binding affinity of the compound or with the additional step of assessing the ability of the compound to mediate hypnotic effects m vivo without causing cognitive impairment via an animal model established to be predictive of sedation and an animal model predictive of cognitive impairment. Thus, the invention provides methods for identifying compounds with hypnotic activity (1) by examining the binding of a given compound at GABA A receptors and (2) by assessing the ability of the compound to potentiate GABA responses at a series of GABA A subtype receptors. The resulting values are then compared to a set of empirically defined criteria termed the "Hypnotic Activity Range Profiler" (Table II, below). The criteria given by the Hypnotic Activity Range Profiler are used to identify compounds with sedative hypnotic activity that have no or very minimal side effects. This activity profile comprises measurements of in vi tro efficacy (agonist, inverse agonist or antagonist character) and EC 50 values at each of 4 GABA A subtype receptors. The activity profile needed for hypnotic effects is presented as a precise window of agonism and EC S0 criteria at each of these subtype receptors. This well-defined activity profile requires partial agonist activity at subtype receptors containing the α ; and α subunits and lower agonist activity at other subtype receptors . The combination of determining efficacy and EC 50 of a test compound is crucial as many compounds bind with high affinity at the benzodiazepine site without potentiating the GABA response at the appropriate subtype receptors . In certain embodiments, the conclusions drawn from the m vi tro determinations may be confirmed by examining the m vivo efficacy of test compounds predicted to have hypnotic properties using animal models for evaluating sedative activity. It may also be desirable to verify that compounds identified by these methods possess the favorable side effect profiles by examining the performance of the compounds in animal models known to be predictive of these side effects. Thus, the animal models may be used as an additional step of the assay to further refine the selection of compounds with selective hypnotic activity. Additional Disclosure: In a further aspect of the present invention, a method of providing pharmaceutical compounds to patients in need of cognition enhancement, hypnosis, anxiolysis, and/or antidepressant treatment (such patients including humans, pets, livestock, and other animals) is provided. In accordance with this method, compounds are obtained that have been identified as having anxiolytic activity, hypnotic activity, antidepressant activity or cognition enhancing activity accordance with any of the novel screening, characterization, analysis or identification methods of the present invention. Preferably such screening, characterization, analysis or identification is carried out outside of the United States of America. Once such compounds have been obtained, they are tested, preferably m vivo, for toxicity and pharmacokmetic properties. At least one compound determined to have minimal toxic effects and to have useful pharmacokmetic properties is then selected for clinical development. By useful pharmacokmetic properties is meant pharmacokmetic properties known m the art to be useful for a compound having the particular activity of anxiolytic activity, hypnotic activity, antidepressant activity or cognition enhancing activity, as identified for each particular compound in accordance with any of the novel screening, characterization, analysis or identification methods of the present invention for each compound. By clinical development is meant those activities, including testing in patients, related to the development and submission of information under a United States Federal law which regulates the manufacture, use, or sale of drugs or veterinary products, such as the Federal Food Drug and Cosmetic Act and other applicable government laws and regulations pertaining thereto. The final step m this method is the offer for sale (preferably m the United States of America) for use as a drug or veterinary product of a pharmaceutical preparation (such as a pill, powder, inhalant, elixir, miectible solution, patch or suppository) comprising the compound . DETAILED DESCRIPTION OF THE INVENTION The invention provides methods for screening compounds for activity as antidepressants, cognitive enhancers, sedative hypnotics, or anxiolytics. Therapeutic compounds identified by these methods mediate effects through the benzodiazepine site of the GABA A receptor without eliciting side effects classically associated with compounds exhibiting such activity that act at this site. Although optional, it is preferred that part b) of each method embodiment (as set forth below) will be conducted on compounds displaying sufficiently potent binding affinities as determined part a) of the method. In one aspect of each embodiment of the method, prior to determining an m vi tro efficacy value for the test compound, the binding affinity of the compound is measured m cells expressing cloned α,β, and γ GABA A receptor subunits or in a cell membrane preparation of such cells. In another embodiment, prior to determining the m vi tro efficacy values for the test compound, the binding affinity of the test compound is determined m any tissue capable of expressing GABA A receptors containing Rol5-1788 binding sites or in a cell membrane preparation of any tissue capable of expressing GABA A receptors containing Rol5-1788 binding sites. In a preferred embodiment of the method, prior to determining in vi tro efficacy values of the test compound, the binding affinity of the test compound is determined in rat spinal cord tissue or in a cell membrane preparation of rat spinal cord tissue. In more preferred embodiments of the method, the binding affinity of a test compound is measured in rat cortex or in a cell membrane preparation of rat cortex. In particularly preferred embodiments, the binding affinity of a test compound is determined in rat cortex or in a cell membrane preparation of rat cortex and the test compound is selected for further evaluation if it gives a K^ value of <100nM or, preferably, <50 nM or, most preferably, < 30 nM. In these embodiments, the binding affinity of a compound may be first determined by evaluating the ability of the compound to displace a radiolabeled compound, for example Rol5- 1788 (Flumazenil) , known to have high affinity at the benzodiazepine site. The in vi tro efficacy and EC B0 value of the test compound may be determined by measuring the chloride flux at the surface of a cell expressing the α,β, and γ subunits of the GABA A receptor m response to GABA by the two electrode voltage-clamp technique. While a variety of cells are suitable for use herein, the preferred cells used m this technique are Xenopus laeviε oocytes that have been injected with non-polyadenylated cRNA coding for human derived α, β and γ GABA A subunits. The preferred form of the γ subunit is the normally expressed long form although an alternatively spliced form may be used. A: Cognitive Enhancers: Cognitive enhancers identified by this method produce inverse agonist activity at the lβ2γ2 or α5β3γ2 GABAA subtype receptors and agonist activity at subtype receptors containing the α.2 or α.3 subunits. Of the possible subunit combinations for receptors containing α.2 or α.3 subunits, the most relevant are the α2β3γ2 and α3β3γ2 subtype receptors. Compounds selected according to the invention have EC S0 values of about 200 nM or less at the αlβ2γ2 and Ct5β3γ2 GABA A subtype receptors and EC 5C values preferably of 150 nM or less at these receptors . The criteria for screening compounds for cognitive enhancing activity are presented below. Table I Cognitive Enhancer Activity Range Profiler *A wide range of EC 50 values for the α 2 β J γ 2 and α 3 β 3 γ 2 subtype receptors is permitted . However , in practice , the "any/ >10% " criteria are used for compounds having EC 50 values at these subtypes lesser than 100 times the EC 50 values at the a^ 2 y 2 or α 5 β 3 γ 2 subtype receptors . On the other hand , when the EC 50 value for a compound at either the ct 2 β 3 γ 2 or the α 3 β 3 γ 2 subtype receptor is more than 100 times the EC 50 values for ^ 2 y 2 or 5 β 3 γ 2 subtype receptors , then <10 % in vi tro eff icacy is acceptable . Thus, methods of this invention comprise: a) screening compounds, optionally ones having a binding affinity less than 100 nM or preferably less than 30 nM at any GABA A receptor; b) determining the m vi tro efficacy and EC 50 values for the compounds at cloned ^ 2 y 2 and α s β 3 γ 2 receptors ; c) determining in vi tro efficacy and EC 50 values for the compounds at GABA A subtype receptors containing the α 2 or α 3 subunit; and d) selecting a compound having an EC 50 value determined in b) of less than 200 nM or preferably less than 150 nM, an efficacy value determined in b) of less than -5% (e.g., -6%, -10%, etc.) or preferably less than -10%, and an efficacy determined in c) of greater than 5% or, preferably, greater than 10%. In preferred embodiments after the binding affinity of the compound has been determined, in vi tro eff icacy and EC 50 values for the test compound are measured at cells expressing the ^ 2 y 2 or a 5 β 3 y 2 GABA A receptor subunit combinations. If the test compound exhibits >5% inverse agonist activity (<-5% efficacy), or preferably >10% inverse agonist activity (<-10% efficacy), at either the α x β 2 γ 2 or the α 5 β 3 γ 2 GABA A receptor and gives EC 50 values of <200 nM, or more preferably <150 nM, at these subunit combinations, these values are compared to in vi tro efficacy values determined using cells expressing GABA A subtype receptors containing α 2 or α 3 subunits. Compounds producing >5% or preferably >10% agonist activity at these subunits are selected as having cognitive enhancing activity. In a preferred embodiment, after the binding affinity of the compound has been determined, in vi tro efficacy and EC 50 value are determined for the test compound using cells expressing the ^ 2 y 2 or a 5 β 3 γ 2 GABA, receptor subunit combinations. If the test compound produces >5% inverse agonist activity (<-5% efficacy), or preferably >10% inverse agonist activity (<-10% efficacy) , at either the α : β 2 γ 2 or α 5 β 3 γ 2 GABA A receptors and gives EC E0 values of <200 nM, or more preferably <150 nM, at these subunit combinations, these efficacy values are compared to the m vitro ef f icacy values determined with cells expressing GABA A subtype receptors containing α 2 β 3 γ 2 or αβ 3 γ 2 subunits. Where the compound produces >5% or preferably >10% agonist activity at these subunits it is selected as having cognitive enhancing activity. In alternative embodiments, the method includes an m v vo evaluation of the ability of the compound to mediate cognitive enhancement without causing proconvulsant effects. This is done using animal models predictive of cognition enhancement and of proconvulεant activity. Compounds that produce a statistically significant effect an animal model predictive of cognitive enhancement are considered to be cognitive enhancing. Compounds that give either a decrease m seizure threshold of less than 25% m the presence of a seizure inducing drug or no significant effect at the p = 0.05 level are identified as lacking proconvulsant activity. In addition, the method may include an evaluation of whether the compound produces anxiogemc effects This is done using an animal model predictive of anxiogenesis . A compound that gives no statistically significant effect the animal model predictive of anxiogenesis is identified as lacking anxiogemc activity. In accordance with another alternative embodiment of the method, the cognitive enhancing properties of the compound are determined without measuring the binding affinity of the compound but with the additional step of measuring the ability of the compound to mediate cognitive enhancement m vivo without proconvulsant effects, via an animal model predictive of cognition enhancement and an animal model predictive of proconvulsant activity. In accordance with yet another alternative embodiment of the method, the cognitive enhancing properties of the compound are determined without measuring the binding affinity of the compound but with the additional step of measuring the ability of the compound to mediate cognitive enhancement m vivo without proconvulsant effects and without anxiogemc effects, via an animal model predictive of cognition enhancement, an animal model predictive of proconvulsant activity, and an animal model predictive of anxiogenesis, respectively. The spatial water maze and step-down passive avoidance models are suitable models for m vivo determinations of cognition enhancement. The bicucullme or PTZ seizure threshold tests are suitable for use m vivo to determine proconvulsant activity. The elevated plus maze model is an example of a model that may be used in vivo to predict anxiogemc activity. B: Hypnotics: Hypnotic compounds identified by this method mediate effects through the benzodiazepine site of the GABA A receptor either without eliciting the side effects classically associated with compounds that act at this site or elicit these side effects only to very low degree. These side effects include cognitive impairment, ataxia, potentiation of alcohol effects, and a tendency for tolerance and drug dependence . More specifically, compounds identified as hypnotics by this method εhow partial agonist activity at ct 2 β 3 γ 2 receptors, stronger partial agonist activity at ct 3 β 3 γ 2 GABA A subtype receptors, and lower activity at subtype receptors containing the a λ or α 5 subunits. Of the possible subunit combinations for receptors containing α : or α 5 subunits the most relevant are the a^ 2 y 2 and α s β 3 7 2 subtype receptors. Additionally, compounds useful for any of these indications must have EC 50 values of 200 nM or less at the c 2 β 3 γ 2 and α 3 β 3 γ 2 GABA ^ subtype receptors and preferably should exhibit EC 50 values of 150 nM or less at theεe receptors. The criteria for selecting a compound as having hypnotic properties are presented below in tabular form. Table II Hypnotic Activity Range Profiler * A wide range of EC 50 values at the αj,β 2 γ 2 and α 5 β 3 γ 2 subtype receptors is permitted in practice, however, the "any/<45%"and "any/<40%" criteria are preferably used for compounds having EC 50 values at these subtypes less than 100 times the EC 50 values at the α 2 β 3 γ 2 and α 3 β 3 γ 2 subtype receptors. On the other hand, when the EC 50 values of the compound at the a^ 2 y 2 or a 5 β 3 γ 2 subtype receptor are greater than 100 times greater the EC 50 values at the α 2 β 3 γ 2 or α 3 β 3 γ 2 subtype receptors, then >45% in vi tro efficacy for the a^ 2 y 2 subtype receptor or >40% in vi tro efficacy for the α 5 β 3 γ 2 subtype receptor is acceptable. Thus in broad aspect, the methods of this invention comprise : a) screening compounds, optionally, selecting compounds having a binding affinity less than lOOnM at any GABA A receptor; b) determining in vi tro efficacy and EC 50 values for the compounds at cloned α 2 β 3 γ 2 and α 3 β 3 y 2 receptors; c) determining in vi tro efficacy and EC 50 values for the compounds at GABA A subtype receptors containing the α : or α 5 subunit; d) selecting a compound having an EC 50 value as determined in b) of less than 200 nM, or preferably less than 150 nM, and an efficacy value for the α 2 β 3 y 2 receptor of greater than 10%, or preferably greater than 20%, an efficacy value for the 3 β 3 γ 2 receptor of greater than 50%, or preferably greater than 60%, an efficacy value for the receptor containing the α : subunit of less than 50%, or preferably less than 45%, and an efficacy value for the receptor containing the α^ subunit of less than 45%, or preferably less than 40%. In preferred embodiments of the invention, after the binding affinity of the compound has been determined, the in vi tro efficacy and EC 50 of the test compound are measured using cells expressing the α 2 β 3 γ 2 GABA A receptor subunit combination and cells expressing the α 3 β 3 γ 2 GABA A receptor subunit combination. If the test compound exhibits EC 50 values of <200 nM or more preferably <150 nM at these subtype receptors, partial agonist activity at 2 β 3 γ 2 receptors and stronger partial agonist activity at α 3 β 3 γ 2 receptors, these values are compared to the in vi tro efficacy and EC 50 values of cells expressing subtype receptors that contain the α : or α s subunits. If the compound also exhibits lower activity at receptors containing cq or α 5 subunits, it is selected as having hypnotic properties. In other preferred embodiments, after the binding affinity of the compound has been determined, the in vi tro efficacy and EC 50 values of the test compound are measured using cells expressing the 2 β 3 γ 2 GABA A receptor subunit combination and cells expressing the α 3 β 3 γ 2 GABA. receptor subunit combinations. If the test compound gives EC S0 values of <200 nM or more preferably <150 nM at these subunit combinations, >10% or preferably >20% agonist activity at the ct 2 β 3 γ 2 subtype receptor, and >50% or preferably >60% agonist activity at the 3 β 3 γ 2 subtype receptor, these values are compared to the in vi tro efficacy and EC 50 values in cells expressing the GABA A receptors containing α 1 or α 5 subunits. If the compound also exhibits <50% or preferably <45% agonist activity at GABA A receptor containing the α x subunit and exhibits <45% or preferably <40% agonist activity at GABA A receptor containing the 5 subunit, it is selected as having hypnotic properties. In more preferred embodiments, after the binding affinity of the compound has been determined, in vi tro efficacy and EC 50 values of the test compound are measured using cells expressing the 2 β 3 γ 2 GABA A receptor subunit combination and cells expressing the α 3 β 3 γ 2 GABA A receptor subunit combination. If the test compound giveε EC C0 valueε <200 nM or more preferably <150 nM at these subunit combinations, >10% or preferably >20% agonist activity at the α_β 3 γ 2 subtype receptor, and >50% or preferably >60% agonist activity at the α j β^ εubtype receptor, these values are compared to the m vitro efficacy and EC S0 values at cells expressing a^ 2 y 2 and α,β 3 . subtype receptors. If the compound also exhibits <50% or preferably <45% agonist activity at the a^ 2 y 2 subtype receptor or exhibits <45% or preferably <40% agonist activity at the α β 2 γ 2 subtype receptor GABA A it is εelected as having hypnotic properties. In alternative embodiments, the method includes measuring the ability of the compound to mediate hypnotic effects m vivo without causing cognitive impairment. This is accomplished using an animal model predictive of a compound's ability to cause hypnotic effects and an animal model developed to be predictive of cognitive impairment. A compound that shows a statistically significant effect m the animal model predictive of sedation and no statistically significant effect in the animal model predictive of cognitive impairment is identified as having hypnotic properties. Suitable m vivo animal models include the εpontaneous locomotor model for predicting hypnotic effects, and the step- down passive avoidance model or the spatial water maze model determining cognitive impairment. C: Anxiolytics: Methods are provided for identifying selective anxiolytic compounds. Therapeutic compounds identified by these methods mediate effects through the benzodiazepine site of the GABA A receptor either without eliciting the side effects classically associated with compounds that act at this site or elicit these side effects only to a very low degree. These side effects include cognitive impairment, sedation, ataxia, potentiation of alcohol effects, and a tendency for tolerance and drug dependence. More specifically, compounds identified as selective anxiolytics by this method show agonist activity at the ct 2 β 3 γ 2 and ct 3 β 3 γ 2 GABA A subtype receptors and lower to no agonist activity at subtype receptors containing the cq or α 5 subunits. Of the possible subunit combinations for receptors containing ct j or α 5 subunits, the most relevant are the a^ 2 y 2 and α s β 3 Y 2 subtype receptors. Additionally, compounds useful for any of these indications must have EC 50 values of 200 nM or less at the α 2 β 3 γ 2 and α 3 β 3 γ 2 GABA A subtype receptors and preferably should exhibit EC 50 values of 150 nM or less at these receptors. The criteria for selecting a compound as having anxiolytic properties are presented below in tabular form. Table III Anxiolytic Activity Range Profiler *A wide range of EC 50 values at the α x β 2 γ 2 and α 5 β 3 γ 2 subtype receptors is permitted. In practice however, the "any/<20%" criteria are preferably used for compounds having EC 50 values at these subtypes lesε than 100 timeε the EC 50 valueε at the α 2 β 3 γ 2 and α 3 p 3 Y 2 subtype receptorε . On the other-hand, when the EC 50 values of a compound at the a^ 2 y 2 or a s β 3 γ 2 subtype receptors are greater than 100 times the EC S0 values at the 2 β 3 γ 2 or 3 β 3 γ 2 subtype receptors, >20% in vi tro efficacy is acceptable. Thus, in a broad aspect the invention comprises: a) screening compounds, optionally compounds having a binding affinity lesε than 100 nM at any GABA A receptor; b) measuring in vi tro efficacy and EC S0 values for the compounds at cloned α 2 β 3 γ 2 and α 3 β 3 γ 2 receptors; c) measuring in vi tro efficacy and EC 50 values for the compounds at GABA A subtype receptors containing the cr^ or α 5 subunit ; and d) selecting a compound having an EC 50 as measured in b) of less than 200 nM and an efficacy value as measured in b) is greater than the efficacy values measured in c. In preferred embodiments, after the binding affinity of the compound has been determined, the in vi tro efficacy and EC 50 of the test compound are measured at cells expressing the α 2 β 3 γ 2 and α 3 β 3 Y 2 GABA A receptor subunit combinations . If the test compound exhibits any agonist activity and gives EC 50 values < 200 nM, or more preferably < 150 nM, at these subunit combinations, these values are compared to the in vi tro efficacies at cells expressing GABA A subtype receptors containing a τ or α 5 subunits. If the compound exhibits lower or no activity at these latter subunits, it is identified as having anxiolytic properties. In other preferred embodiments, after the binding affinity of the compound has been determined, the in vi tro efficacy and EC S0 of the test compound are measured using cells expressing the α 2 β 3 γ 2 and α 3 β 3 γ 2 GABA A receptor subunit combinations. If the test compound exhibits any agonist activity and gives EC SC values of < 200 nM, or more preferably < 150 nM, at these subunit combinations, these values are compared to the in vi tro efficacy in cells expressing the ^ 2 y 2 GABA A receptor subunit combination. If the compound exhibits lower to no activity at this subunit it is identified as having non-sedating anxiolytic properties. In further embodiments, after the binding affinity of the compound has been determined, the in vi tro efficacy and EC 50 of the test compound are measured using cells expressing the α 2 β 3 γ 2 and α 3 β 3 γ 2 GABA A receptor subunit combinations. If the test compound exhibits agonist activity and gives EC S0 values of < 200 nM, or more preferably < 150 nM, at these subunit combinations, the efficacy values are compared to in vi tro efficacy values using cells expressing the a^ 2 y 2 and α 5 β 3 γ- GABA A receptor subunit combinations. If the compound exhibits lower to no activity at these subunits it is identified as having non-sedating anxiolytic properties. In still other embodiments, after the binding affinity of the compound has been determined, in vi tro efficacy and EC 50 values for the test compound are measured cells expressing the α 2 β 3 γ 2 and α 3 β 3 γ 2 GABA A receptor subunit combinations. If the test compound gives >30% potentiation (i.e., increase) of the GABA response and EC S0 values of < 200 nM, or more preferably < 150 nM at these εubunit combinations, the efficacy values are compared to the m vi tro efficacies determined using cells expressing GABA A subtype receptors containing - or α t subunits. If the compound exhibits lower to no activity (efficacy) at these subunits it is identified aε having non-sedating anxiolytic properties. In yet other embodiments, after the binding affinity of the compound has been determined, m vi tro efficacy and EC 50 values for the test compound are measured using cells expressing the α 2 β 3 γ 2 and α 3 β 3 γ 2 GABA A receptor subunit combinations. If the test compound produces >30% potentiation of the GABA response and EC εo values of < 200 nM, or more preferably < 150 nM, at these subunit combinations, these efficacy values are compared to the m vi tro efficacy values determined using cells expressing the α-iβ ∑ Y ∑ GABA A receptor subunit combination. If the compound exhibits lower to no activity efficacy at this subunit, it is identified as having non-sedatmg anxiolytic properties. In more preferred embodiments, after the binding affinity of the compound has been determined, m vi tro efficacy and EC S0 values for the test compound are measured using cells expressing the α 2 β 3 γ 2 and α 3 β 3 γ 2 GABA A receptor εubunit combinations. If the test compound produces >30% potentiation of the GABA response and EC 50 values of < 200 nM, or more preferably < 150 nM, at these subunit combinations, these values are compared to the in vi tro ef f cacy values measured with cells expressing the ct.,β 2 γ 2 and α 5 β 3 γ 2 GABA A receptor subunit combinations. If the compound produces lower to no efficacy activity at these subunits, it is identified as having non-sedatmg anxiolytic properties. In particularly preferred embodiments, after the binding affinity of the compound has been determined, in vi tro efficacy and EC 50 values for the test compound are measured using cells expressing the α 2 β 3 γ 2 and α 3 β 3 γ 2 GABA Λ receptor εubunit combinations. If the test compound produces > 30% potentiation of the GABA response and EC 50 values of < 150 nM at these subunit combinations, the efficacy values are compared to m vi tro efficacy measured using cells expressing the α_β 2 γ 2 and α 5 β 3 y 2 GABA A receptor subunit combinations. If the compound gives < 20% potentiation of the GABA response at these latter subunits, it is identified as having non-sedatmg anxiolytic properties. In alternative embodiments, the method includes the additional step of measuring the ability of the compound to mediate anxiolytic effects m vivo without causing sedation. This is accomplished using animal models established to be predictive of anxiety and sedative effects. A compound that shows a statistically significant effect in the animal model predictive of anxiety and no statistically significant effect in the animal model predictive of sedative effects is identified as having non-sedatmg anxiolytic properties. Suitable in vivo animal models include the elevated plus -naze model for predicting anxiolytic activity and the spontaneous locomotor activity model to determine sedative effects . Antidepressants : A method lε provided for identifying antidepreεsant compounds. Therapeutic compounds identified by this method act through the benzodiazepine site of the GABA A receptor without eliciting the side effects classically associated with compounds that bind at this site. Alternatively, these compounds elicit the side effects only to a very low degree. These side effectε include cognitive impairment, sedation, ataxia, potentiation of alcohol effects, and a tendency for tolerance and drug dependence. More specifically, compounds identified as antidepressants by this method show agonist activity at the 2 β 3 γ 2 and α 3 β 3 γ 2 GABA^ subtype receptors and lower or no agonist activity at subtype receptors containing the α,_ or α 5 subunits. Of the possible subunit combinations for receptors containing o^ or a 5 εubunitε, preferred for use herein are the cι. 1 β 2 y_ and α ζ β 3 γ, subtype receptors . Compounds useful for any of these indications must produce EC 50 values of 200 nM or less at the α 2 β 3 γ 2 and α 3 β 3 γ >: GABA A subtype receptors and preferably should exhibit EC 5C values of 150 nM or lesε at these receptors. The preferred criteria for screening for compounds having antidepressant activity are presented m the table below. Table IV Antidepressant Activity Range Profiler *A wide range of EC 5C values for the a^ 2 y 2 and s β 3 γ 2 subtype receptors is permitted. In practice, however, the "any/<20%" criteria are used for compounds having EC 50 values at these subtypes less than 100 times the EC ςo values at the α 2 β 3 γ 2 and α 3 β 3 γ 2 subtype receptors. On the other hand, when the EC 50 value for the compound at either the ct^ 2 y 2 or the α 5 β 3 y 2 subtype receptor is greater than 100 times the EC 50 values for either the α 2 β 3 γ 2 or the α 3 β 3 Y 2 subtype receptors, then >20% m vi tro efficacy is acceptable . Thus, the method of this invention comprises : a) screening compounds, optionally compounds having a binding affinity lesε than 100 nM at any GABA A receptor; b) determining m vi tro efficacy and EC 50 values for the compounds at cloned 2 β 3 γ 2 and 3 β 3 γ 2 receptors; c) determining m vi tro efficacy and EC 50 values for the compounds at GABA A subtype receptors containing an . or α 5 subunit; and d) selecting a compound having an EC C „ as determined in b) of less than 200 nM and an efficacy value as determined b) greater than the efficacy value measured c. In preferred embodiments, after the binding affinity of the compound has been determined, m vi tro ef f icacy and EC 50 values for the test compound are measured using cells expressing the α 2 β 3 γ 2 and 3 β 3 γ 2 GABA. receptor εubunit combinations . If the test compound exhibits any agonist activity and produces EC S0 values of < 200 nM or, more preferably, < 150 nM at these subunit combinations these values are compared to in vi tro efficacy values determined using cells expressing GABA A subtype receptors containing α or α c subunits . Compounds giving lower or no efficacy activity at the α or α c subunits are selected as having antidepreεεant propertieε. In a preferred aspect, the in vi tro efficacy and EC 50 values are determined for the test compound using c β-f anc i o^β s Y ∑ GABA A receptor subunit combinations. If the test compound exhibits agonist activity and EC C0 values < 200 nM or more preferably < 150 nM at these subunit combinations, these values are compared to m vi tro efficacy values determined with cells expressing the α-iβ ∑ Y ∑ GABA A receptor εubunit combination. Where the compound exhibits lower or no efficacy activity at the a^ 2 y 2 subunit, it is identified as having antidepressant properties. In more preferred aspect, the m vi tro efficacy and EC 50 values determined for the test compound using cells expressing the ot 2 β 3 γ 2 and ct 3 β 3 γ ^ GABA A receptor subunit combinations are compared to m vi tro efficacy values determined using cells expressing the α j β 2 γ 2 and α s β 3 γ 2 GABA A receptor subunit combinations combination. Where the compound exhibits lower to no efficacy activity at both the a^Y 2 and α s β 3 γ 2 subunits, it is identified as having antidepressant properties. In these embodiments, agonist activity is preferably defined as producing an efficacy value of >30% potentiation of the GABA responεe. Thus, m preferred embodiments of the invention if the test compound exhibits >30% potentiation of the GABA response at the α 2 β 3 γ 2 and α 3 β 3 γ ^ receptor subunits, and EC S0 valueε of < 200 nM or, more preferably < 150 nM, at these subunit combinations, these values are compared to the m vi tro efficacy at cells expressing the α,β 2 γ 2 GABA A receptor subunit combination. Where the compound exhibits lower to no efficacy (agonist) activity at the ^ 2 y 2 subunit, it is identified as having antidepressant properties. Particularly preferred embodiments comprise- determining the binding affinity of the compound for GABA A receptors determining in vi tro efficacy and EC 50 values for the test compound with cells expressing the 2 β 3 γ 2 and α 3 β 3 γ 2 GABA A receptor subunit combinations. Where the test compound produces >30% potentiation of the GABA response (i.e., agonist activity) and EC S0 values of < 200 nM or, more preferably < 150 nM, at these subunit combinations, these efficacy values are compared to m vitro efficacy values determined using cells expressing the ct^ 2 y 2 and α c β 3 γ 2 GABA A receptor subunit combinations. Where the compound exhibits < 20% potentiation of the GABA response at the a^ 2 y 2 and ct ζ β 3 γ 2 subunits, it is identified as having antidepressant properties . In alternative embodiments, the method includes, an m vivo evaluation of the ability of the compound to mediate antidepresεant effects without causing sedation. This is done using animal models predictive of antidepressant activity and sedation. A compound that produceε a statistically significant effect in an animal model predictive of antidepressant activity and no statistically significant effect m an animal model predictive of sedative effects is identified as having antidepressant properties. Suitable m vivo animal models include the Porsolt swim test for predicting antidepresεant activity and the εpontaneous locomotor activity model for determining sedative effects. In the forgoing embodiments and in the claims, any of the various criteria presented for characterizing each of hypnotic, antidepressant, anxiolytic or cognition enhancing properties (including those set forth in the various dependent claims submitted herewith regarding a particular one of such properties) may be applied in association with each aspect or embodiment of the invention concerning the characterization of that particular property and to each of the various independent claims submitted herewith regarding the characterization of that particular property. The methods of the present invention are illustrated further by the following examples, which are not to be construed as limiting the invention m scope or spirit to the specific procedures and compounds described them. EXAMPLES Example 1. BINDING ASSAYS A preferred method for discovery of compounds that bind with high affinity to GABA A receptors is a competition binding assay. Rat cortex membranes are prepared by homogenizing one or two previously frozen rat cortexes m 30 ml of 25 mM Tris Buffer, pH 7.4. The homogenate is centrifuged for 10 minutes at 500 x g. The supernatant is then transferred to a clean centrifuge tube and the pellet discarded. The supernatant is spun for 20 minutes at 48,000 x g. The supernatant from this spin is discarded and the pellet is resuspended in 30 ml of 25 mM Tris Buffer, pH 7.4 and centrifuged for an additional 10 minutes at 48,000 x g. The supernatant from the final spin is discarded and the membrane pellet is resuεpended in 100 mL Tris buffer per gram of cortex used. The cortex membrane preparation is used to perform either percent inhibition or competition binding assays . In order to determine percent inhibition 300 μl of reεuspended membranes are mixed with 200 μl 3 H labeled Rol5-1788 (final concentration 2.5 nM) and incubated for 1 hour on ice m the presence of 2 μl test compound in DMSO (final concentration 4 μM) . Membranes are harvested onto untreated filtermatε. The filtermatε are dried and the H Rol5-1788 signal lε counted m a scintillation counter. Nonspecific binding is determined by displacement of 3 H Rol5-1788 with 10 μM Diazepam (RBI) or any other compound known to bind tightly at the benzodiazepine site. For any test compound exhibiting a favorable percent inhibition a competition binding curve is obtained and a K 1 value is calculated. Generally, up to 11 points spanning the compound concentration range from 10 12 M to 10 B M are obtained per curve by the same method as for the percent inhibition assay. K values are calculated according to the Cheng-Prusεof equation. Thoεe compounds that exhibit Ki values m the desired range are submitted for efficacy testing. Example 2. ELECTROPHYSIOLOGY ASSAYS The efficacy profile of compounds of thiε invention is determined by the following electrophysiological assay for GABA A receptor activity. Assays are carried out as described previously m White and Gurley, 1995 and White, et al . , 1995. Xenopuε laevis oocytes are enzymatically isolated and injected with non-polyadenylated cRNA for human derived α, β and γ GABA t subunits, respectively. In more preferred embodiments cRNA for the ^ 2 y_ c β . . a 3 β 3 γ 2 and α 5 β 3 γ ^ subunit combinations are injected. Only one of these subunit combinations is injected per cell. For each subunit combination, sufficient meεεage is injected to result current amplitudes of >10 nA when 1 μM GABA is applied. Electrophysiological recordings are carried out using the two electrode voltage-clamp technique at a membrane holding potential of -70 mV. Compounds are evaluated against a GABA concentration that evokes <10% (EC,„) of the maximal evocable GABA current. Each oocyte is exposed to increasing concentrations of compound in order to evaluate a concentration/effect relationship. Compound efficacy is expressed as a percent-change m current amplitude: 100* ( (Ic/I) -1) , where Ic is the GABA evoked current amplitude observed m the presence of compound and I is the GABA evoked current amplitude observed in the absence of compound. Specificity of a compound for the Rol5-1788 site is determined following completion of the concentration/effect curve. After washing the oocyte sufficiently to remove previously applied compound, the oocyte is exposed to GABA and 1 μM Rol5-1788, followed by exposure to GABA, 1 μM Rol5-1788, and compound. Percent change due to addition of compound is calculated as described above. Any percent change observed in the presence of Rol5-1788 is subtracted from the percent changes m current amplitude observed m the absence of 1 μM Rol5-1788. These net values are used for the calculation of average activity and EC EC values. To evaluate average activity and EC 5C values, the concentration/ effect data are averaged across cells and fit to the logistic equation. Average values are reported as mean ± standard error. In the preferred embodiment, anxiolytic compounds should exhibit an activity profile of <20% agonist activity at the α,β 2 γ 2 and α 5 β 3 γ 2 subunit combinations and >30 % agonist activity at the α 2 β 3 γ 2 and α 3 β 3 γ 2 εubunit combinations and EC 50 values < 150 nM. Preferred hypnotic compounds exhibit an activity profile of >20% agonist activity at the α 2 β 3 γ 2 subtype receptor, >60% agonist activity at the α 3 β,γ 2 subtype receptor, <45% agonist activity at the o^β^ subtype receptor, and <40% activity at the α 5 β 3 γ 2 subunit combinations and EC S0 values of <150 nM at the α 2 β 3 y 2 and α 3 β 3 γ 2 constructε . Preferred antidepressant compounds exhibit an activity profile of <20% agonist activity at the α,β 2 γ 2 and α^β ^ γ- subunit combinations, >30 % agonist activity at the α 2 β 3 γ 2 and α 3 β J γ ^ subunit combinations, and EC 50 values of <150 nM at the α,β 3 γ 2 and α 3 β 3 γ 2 constructs. Preferred cognitive enhancing compounds produce an efficacy profile of >10% inverse agonist activity (<-10% efficacy) at the ct^ 2 y 2 or α 5 β 3 γ 2 subtype receptors, >10 % agonist activity at the a 2 βf/ 2 and α 3 β 3 γ 2 subunit combinations, and produce EC 5C values of < 150 nM at the ,β 2 γ 2 and α 5 β 3 γ 2 subtype receptors. The modulatory effect on the GABA current amplitude is near maximal for all subtype receptors studied. Example 3. ANIMAL BEHAVIOR METHODS To verify that compoundε identified as selective anxiolytics, hypnotics, antidepresεantε , or cognitive enhancers by this method mediate these effects it is preferable to examine the performance of some such compounds m vivo . A εeπes of animal models, most preferably rat models, are employed for this purpose . It has been established that the elevated plus maze model can be used to determine anxiolytic efficacy of compounds. Side effects asεociated with anxiolytic compoundε can be ascertained from a variety of animal models . The spontaneouε locomotor activity model can be uεed to determine whether test compounds cause sedation. Spontaneous locomotor activity has been established as a measure of hypnotic effect. Among the most common side effectε of currently preεcribed εedative hypnoticε are learning and memory deficits or cognitive impairmen . Consequently, one of the goals of the sedative hypnotic program has been to develop a sedative hypnotic compound with minimal side effects of this nature. The step-down passive avoidance and spatial water maze paradigms can be used to determine whether test compounds cause learning and memory deficits. The Porsolt swim test has been established as a meaεure of determining antidepressant activity of compounds. Side effects associated with antidepressant compounds can be ascertained from a variety of animal models. For example, the spontaneouε locomotor activity model may be used to determine whether test compounds cause seαation. The spatial water maze model or the step-down passive avoidance model may be used to measure cognitive enhancement. Proconvulsant activity and anxiogemc effects are the side effects of greatest concern for compounds that act as cognitive enhancers via the benzodiazepine site. The bicucullme εeizure threshold or the PTZ seizure threshold testε may be used to determine whether cognition enhancing compounds are proconvulsant, the elevated plus maze may be used to determine whether such compounds are anxiogemc. The aosence of side effectε for a compound that shows m vivo efficacy is indicative that the compound not only has the desired activity but that it is also a selective compound. A statistically significant effect m the spatial water maze, step-down passive avoidance, spontaneous locomotor activity, Porsolt swim test, or the elevated plus maze models is defined as p <0.05 using a valid parametric statistical test. Likewise, no statistically significant effect is defined these models as p >0.05 using a valid parametric statistical test. No statistically significant effect for either of the seizure threshold models is defined as a decrease of less than 25% in the seizure threshold the presence of a seizure inducing drug or a result that is not significant at the p < 0.05 level. a. Elevated plus maze The elevated plus maze model capitalizes on rats' innate fear of open, elevated places. The test apparatuε is an elevated plus maze consisting of two open arms and two closed arms. A rat will naturally chose to spend more time on the closed arms of the maze than on the open arms but that if an efficacious anxiolytic compound is administered to the rat prior to the test the amount of time the rat spendε m the open armε is increased. Test compound is administered IV m a 50% PEG (polyethylene glycol) vehicle 5 minutes prior to the test session. A range of compound doses is typically used and 8 - 10 rats are tested at each dose. The rat is placed m the center of the maze facing one of the open arms. The animal's locomotion is tracked over a five minute test seεεion using photocellε interfaced to a computer. The computer measures the number of entries into each arm and the time spent on each arm. An anxiolytic effect m the elevated plus maze model is defined by an increase m the percentage of time spent on the open arms m compound versus vehicle treated animals. b . Spontaneous Locomotor Activi ty- Spontaneous locomotor activity can be measured to determine εedative effects of compounds. Locomotor activity is measured in eight computerized Dιgιscan-16 Animal Activity Monitors (Omnitech Electronics, Columbus, Ohio) equipped with 48 infrared photocell emitters and detectors . Each box is constructed of Plexiglas sides and floor. Horizontal activity is detected by a set of horizontal sensors on the front to back walls and a second group of senεorε on the εide to side walls located 5 cm above the cage floor. Vertical activity is detected by a third set of εenεors on the εide to εide walls located 13.5 cm above the cage floor. The rats are tested m the presence of white noise (62 dB) and red light (60 watt) . Test compound is administered IV m a 50% PEG vehicle 5 minutes prior to the test session. A range of compound doses is typically uεed and 6 to 8 rats are tested at each dose. The animal's movement time, vertical activity, and total distance traveled are tracked over a 15 minute test session. A sedative, or hypnotic, effect m the spontaneous locomotor activity model is defined by a decrease m any two of these three measures relative to animals given vehicle alone. c. Step-Down Passi ve Avoidance In step-down pasεive avoidance a rat lε placed on a platform located m the center of an electrified grid floor that lε contained within a large (45 cm x 45 cm x 50 cm) white tranεlucent Plexiglas * enclosure. The natural inclination of the rat is to εtep off the platform and investigate its surroundings . In day one of the experiment animals are treated with either Zolpide , test compound m a 50% PEG vehicle, or vehicle alone and then trained to remain on the platform for at least 120 secondε . Each time the animal steps off the platform it receives a mild foot shock of 2 mAmps x 6 εec . Following each shock the animal is removed from the box, placed its cage for a one minute mter-trial interval, and then returned to the platform. The latency to step down on each trial, the number of trials taken to reach criterion during training and the retention latency are collected. Testing is conducted approximately 24 hrε . after training. Drug- free animals are placed on the platform m the box m which they will have been trained and the latency to step down onto the grid floor is recorded for one trial as a measure of memory retention. The animal is allowed a maximum of 120 seconds to step down and does not receive a shock upon stepping off the platform. d. Porsol t Forced Swim Test The effects measured m this model have been correlated to antidepreεεant efficacy for drugε . The paradigm of this model is that an effective antidepressant compound will cause a rat to make greater attempts to escape a water- filled cylinder than a rat given vehicle only. Animals used m thiε study were non-naive male Sprague Dawley Rats (SASCO, St. Louiε) weighing between 280 - 350 grams. The teεt apparatuε consists of 6 clear Plexiglas cylinders 40 cm high x 19 cm wide Cylinders are filled to 18 cm with 25 °C water. Each rat is placed m a cylinder for a 15 minute training session. Following either subchronic or acute dosing of either vehicle (0.5% methylcelluloεe) of compound, animals are brought back 24 hours later for a 5 minute test session. These test session is videotaped for later scoring. Subchronic dosing consists of administering drug three times m the 24 -hour period between training and testing. The drug is administered 24 hrε . , 5 hrε., and 1 hr . prior to the teεt seεsion. Acute dosing consists of administering the drug once, 1 hour prior to the test εeεεion. Scoring lε done using a time-sampling computer program written m Visual Basic and run in DOS. Every five seconds, animals are rated aε demonstrating one of three behaviors. immobility, mild swim, or climbing. These sampling score are then converted into percentages of the test seεεion. e. Spatial Water Maze The spatial water maze has been used extensively as a test of spatial learning and memory. Rats are trained to escape from the water by swimming to a platform that is submerged uεt below the εurface of the water. Since the platform is not visible to the animal, it has to utilize visual extra-maze cues the area of the tank to locate the platform. The water maze apparatus consists of a circular tank, 119 cm m diameter and 56 cm m height, with a black interior. The tank is filled with water approximately 23 - 25 °C to a height of 42 cm. Superimposed onto the tank are four quadrants, South, East, North and West The tank is surrounded by external visual cues, which consiεt of a black and white checkered wall, a black and white εtπped wall, a white wall with two light fixtureε, and a blue wall. A black circular PLEXIGLAS platform with a plack neoprene rubber top lε placed the Northeaεt quadrant approximately 1 - 2 cm below the surface of the water. The submerged platform is 39 cm m height and haε a diameter of 11.5 cm. Training and testing are conducted m the presence of a 60 - 62 dB white noise source and under dim light conditions (1.0 - 1.2 lux) . The animal's path is tracked by a video camera interfaced to an automated tracking package (Videotrack, CPL Systems) . Acquisition Training. Training consists of six trials. Test compound is administered IV m a 50% PEG vehicle 5 minutes prior to the test session. A range of doses is typically used and 8 to 10 rats are teεted at each dose. Each animal is placed on the platform m tne tank for 20 seconds prior to the first trial of acquisition training For the first trial, the animal is placed m the water facing the wall of the tank at the "South" starting position. The order of training trials is South, East, North, and West. Eacn of the training trials is separated by an mter- trial interval (ITI) of 3 minutes. Each trial ends with the animal finding the platform or being placed onto it after 90 sec. Rats are then given 10 secondε on the platform and removed from the maze for the ITI . Each trial endε with the animal finding the platform or being placed onto it after 90 seconds. Rats are then given 10 seconds on the platform and removed from the maze for the ITI. During the ITI, each rat is dried off with a towel and placed m a heating chamber maintained at 45 °C. The latency to reach the submerged platform (measured m secondε) , the total diεtance traveled m the maze (meaεured in meterε) , the number of zone or quadrant transitions made, and the swim speed (meaεured m meters/ sec.) on each trial are all recorded. Upon completion of training animals are returned to their home cages m the vivarium. Retention Testing: Approximately 24 hours after training each rat is tested for retention on one trial. The rat is placed the "South" starting position and given 90 seconds to located the platform. The latency to locate the platform, total distance traveled, number of zone transitions and swim speed are all recorded by computer. f . Bicucullme Seizure Threshold Test . This teεt provideε a meaεure of whether the test compound is proconvulsant by measuring whether the test compound produces any change m the dose of the seizure inducing drug, bicucullme, needed to elicit seizures in rats. Test compounds must show <25% decrease m seizure threshold or p >0.05 significance to be considered as drug candidates. Adult male Sprague-Dawley rats (175 - 300 g) are weighed and placed in a rat restrainer. A Teflon indwelling catheter is placed m one of the lateral tail veins and held in place with surgical tape. The catheter and a 3.0 cc syringe are connected by a length of PE-100 tubmg fitted with Hamilton male and female adapter. Patency of catheter placement is tested by backflow of venous blood and ready infusion of lesε than 2 ml of saline solution. Animals are placed m a clear acrylic cage for behavioral observation. The test compound is prepared m either 25% or 50% polyethylene glycoi 400 (PEG) vehicle. A bicucullme (BIC) stock solution of 1.0 mg/ml BIC m 25% PEG/ 75% IN HC1 is also prepared Immediately prior to infusion 0.5 ml of BIC stock is diluted to 20 ml with 19.5 ml of ice cold saline. Test compound is infused 5 minutes prior to the start of bicucullme infusion. Injection volumes do not exceed 4.0 ml/ kg and are typically 1.0 or 2.0 ml/ kg. The catheter lε flushed immediately with 2 0 ml of saline to ensure total delivery of the drug. The catheter is then preloaded with dilute pontamme skye blue dye followed by some air to allow obεervation of the initiation of the bicucullme infusion. The BIC solution is delivered at a speed of 2.0 ml/ minute by a constant speed infusion pump. The final infusion rate of bicucullme is approximately 0.05 mg/ minute. The time elapsed from the start of bicucullme infusion to firεt myoclomc erk (first head/ neck twitch) , the initiation of full myoclonus (writhing) , and final forelimb extenεion is all recorded using a digital stopwatch. Animals are subsequently euthanized The bicucullme seizure threshold is defined as the amount of bicucullme required to titrate the first myoclomc jerk. Bicucullme seizure threshold is expresεed m mg/ kg aε follows : [t me of myoclomc lerk (minutes) 1 x [infusion rate (mg/ minute) ] [weight of animal (kg) ] g. Pentelenetetrazol Seizure Thresnold Test This assay may be used as an alternative to the bicucullme seizure threshold teεt to determine whether a teεt compound is proconvulsant Pentelenetetrazol (PTZ) is used to induce seizures rather than bicucullme. Compound is administered 5 minutes prior to infusion of PTZ as m the bicucullme seizure threshold test. PTZ (2.5 mg/ ml in 0.09% NaCl) is infused at a constant rate of 1.92 ml/ minute with a syringe pump or a constant drug delivery rate of 4 8 mg/ minute The time elapsed from the start of PTZ infusion to first myoclomc jerk (first head/ neck twitch) , the initiation of full myoclonus (writhing) , and final forelimb extension is recorded using a digital stopwatch. The PTZ seizure threshold is defined as the amount of infused PTZ required to titrate the first myoclomc jerk and is calculated by the same relationship as used for the bicucullme seizure threshold In either the bicucullme or the PTZ seizure threshold test compounds should decrease the seizure threshold by lesε than a 25% decreaεe or not εhow a significant effect at the p <0.05 level. Example 4 ANXIOLYTIC COMPOUNDS The compounds listed m Table V were tested by each of the methods described above for identifying anxiolytics compounds and compared to the known anxiolytic Alprazolam. Alprazolam is known to cause side effects including sedation. Table V ' For behavioral assays results are given aε the minimal efficaciouε dose of compound in mg/ kg, administered IV, needed to elicit a statistically significant response. * NS : no statistically significant effect was obεerved over the dose range studied. The structures of the above compounds are given in Table VI Example 5. HYPNOTIC COMPOUNDS The compounds listed in Table VII were tested by the methods described above for identifying hypnotic compounds and compared to the known hypnotic Zolpidem. Zolpidem is known to display side effects including cognitive impairment. Table VII ♦ Minimal efficacious dose of compound, administered IV, needed to elicit a statistically significant response (mg/kg) . T NS indicates that no statistically significant effect was observed this assay over the dose range studied. ***No meaningful value obtained. The structures of Compounds 6-9 are shown m Table VIII, Example 6 ANTIDEPRESSANT COMPOUNDS The compounds εhown in Table IX were tested by each of the methods described above and compared to the antidepressant compound Rol6-6028. ' Minimal efficacious dose of compound , administered IV, needed to elicit a statistically significant response (mg/kg) . * NS indicates that no statistically significant effect was observed this assay over the dose range studied. The compounds shown m Table IX also met the criteria for anxiolytic compounds The structureε for these compounds are given m Table VI, above Example 7 COGNITION ENHANCING COMPOUNDS The following compoundε were teεted by the methods described above for identifying cognition enhancing compounds and compared to CGS 8216, a compound that has been εhown to have a positive effect in models of learning and memory. We have also shown CGS 8216 to be efficacious m the step-down passive avoidance model (0.06 mg/ kg minimal efficacious dose). Additionally it is know that CGS 8216 is anxiogemc and proconvulsant . Table X τ The cognitive model is the step-down passive avoidance model. The result is given as the minimal efficacious dose of compound in mg/ kg, administered IV. " The cognitive model is the spatial water maze model. For the spatial water maze asεay, reεults are given as the minimal efficaciouε doεe of compound m mg/ kg, administered IV (unlesε listed as PO) , needed to elicit a statistically significant response . 'Mean percent decrease in seizure threshold over the dose range studied. * NS indicates that no statistically significant effect was observed in this assay m compound versus vehicle treated animals over the dose range studied. Structure s f or Compoundε 10 - 13 above are preεented in Table XI The foregoing describes preferred embodiments of the present invention. Those of skill in the art will recognize that modifications may be made therein without departing from the spirit or scope of the present invention as set forth in the following claims, which conclude this specification.

Claims: What is claimed is : 1. A method for screening a plurality of compounds so as to identify at least one compound exhibiting cognitive enhancing activity, comprising: a) determining m vi tro efficacy and EC S0 values for each compound at an α,β 2 γ 2 or an 5 β 3 γ 2 GABA. subtype receptor; b) determining an in vi tro efficacy value for each compound at a GABA A receptor comprising an α or α 3 subunit, and c) identifying as exhibiting cognitive enhancing activity a compound having: an EC S0 value determined m a) of lesε than about 200nM, an efficacy value determined in a) of less than about -5%, and an efficacy value determined in b) of greater than about 5%. 2. The method of Claim 1 wherein the EC 50 meaεured m step a) is leεε than 150 nM. 3. The method of Claim 2 wherein the in vi tro efficacy measured at said a^ 2 y 2 GABA A subtype receptor or said α 5 β 3 γ 2 GABA A subtype receptor is less than -10%. 4. The method of Claim 3 wherein the m vi tro efficacy measured at said GABA A receptor comprised of said α 2 subunit or said α subunit is greater than 10%. 5. The method of Claim 1 wherein the m vi tro efficacy measured at said a^ 2 y 2 GABA A subtype receptor or said α s β 3 γ 2 GABA. subtype receptor is less than -10%. 6. The method of Claim 5 wherein the m vi tro efficacy measured at said GABA. receptor comprised of said α 2 or said α, subunit is greater than 10%. 7. The method of Claim 1 wherein the GABA A receptor comprised of said α 2 subunit lε an α 2 β 3 γ 2 GABA A receptor or the GABA A receptor comprised of said α, subunit is an α 3 β 3 γ 2 GABA A receptor. 8. A method for screening compounds for cognitive enhancing activity, comprising: a) selecting compounds having a binding affinity less than 100 nM at any GABA A receptor; b) determining m vi tro efficacy and EC 5C values for each selected compound at an a. 2 f 2 or α 5 β 3 γ 2 GABA A subtype receptor; c) determining m vitro efficacy and EC 50 values for each selected compound at a GABA. receptor comprised of an α 2 or α, subunit; and d) identifying as having cognitive enhancing activity any compound having an EC CC value determined in b) of less than 200nM and an efficacy value measured in b) of less than -5%, and an efficacy value measured in c) of greater than 5%. 9. A method of providing a pharmaceutical preparation to patients m need of cognition enhancing treatment comprising: a) obtaining at least one compound identified as exhibiting cognition enhancing activity by the method of Claim 1; b) testing said at least one compound and submitting results of said testing as part of submission of information under a United States Federal law which regulates the manufacture, use, or sale of drugs or veterinary products; c) showing a pharmaceutical preparation comprising said at least one compound to be safe for use as required by the provisions of the Federal Food Drug And Cosmetic Act; and d) offering the pharmaceutical preparation for sale in the United States of America for use as a cognition enhancing drug or cognition enhancing veterinary product . 10. A method for screening a plurality of compounds for cognitive enhancing activity, comprising: a) determining m vi tro efficacy and EC 50 values for each compound at a^ 2 y 2 or a 5 β 3 γ 2 GABA A receptors ; b) determining m vi tro efficacy for each compound at a GABA A receptor comprised of an α 2 or α 3 subunit; c) determining the in vivo effect of each compound in an animal model for measuring cognitive enhancement; d) determining the in vivo effects of each compound in an animal model for proconvulsant activity by measuring a seizure threshold in the presence of a seizure inducing compound or m an animal model that predicts anxiogemc effects, and e) identifying a cognitive enhancing compound as a compound having cognitive enhancing properties when the EC 50 measured in step a) is lesε than 200nM and the efficacy meaεured m εtep a) is lesε than -5% and the efficacy meaεured m εtep b) is greater than 5% and said compound produces a statistically significant (p <0.05) positive effect m the animal model indicative of cognitive enhancement and said compound does not produce an effect in the animal model predictive of proconvulsant activity of more than a 25% decrease in the seizure threεhold m the presence of the seizure inducing drug, or does not produce a change that is statistically significant said model, or the compound does not produce a statistically significant effect in the animal model that predicts anxiogemc effects . 11. A method for screening compounds for cognitive enhancing properties, comprising: a) selecting compounds having binding affinities of less than 100 nM at any GABA A receptor; b) meaεuring the m vi tro efficacy of each compound at an a^ 2 y 2 or 5 β 3 γ 2 GABA. receptor; c) meaεuring the in vi tro efficacy of each compound at a GABA A receptor compπεed of an α 2 or α, εubunit; d) meaεuring the m vivo ef f ect of each compound in an animal model predictive of cognitive enhancement; e) meaεuring the in vivo side effects of each compound in an animal model that predicts proconvulsant activity by meaεuring a εeizure threεhold in the presence of a seizure inducing compound or measuring the in vi vo side effects of each compound in an animal model that predicts anxiogemc effects; and f) identifying as a cognitive enhancing compound a particular compound for which the EC 5C meaεured m step b) is leεs than 200nM and the efficacy meaεured m step b) is leεε than -5% and the efficacy meaεured in step c) is greater than 5% and said particular compound produces a statistically significant (p <0.05) positive effect in the animal model indicative of cognitive enhancement and said particular compound does not produce an effect in the animal model predictive of proconvulsant activity of more than a 25% decrease in the seizure threshold in the presence of the seizure inducing drug, or does not produce a change that is statistically significant in said model, or said particular compound does not produce a statistically significant effect in the animal model that predicts anxiogenic effects. 12. A method for screening compounds for hypnotic activity, comprising: a) determining EC 50 and in vitro efficacy of each compound at an α 2 β 3 γ 2 GABA. subtype receptor or an α 3 β 3 γ 2 GABA A subtype receptor; b) determining in vi tro efficacy of each compound at a GABA A receptor comprised of an α x or α 5 subunit; and c) selecting a compound having an EC S0 determined in a) of less than 200nM, an in vi tro efficacy determined in a) of greater than 10% for said α 2 β 3 γ 2 GABA A subtype receptor or greater than 50% for said 3 β 3 γ 2 GABA A subtype receptor; and an in vitro efficacy value determined in b) of lesε than 50% for the GABA A receptor comprised of an α subunit or lesε than 45% for the GABA A receptor comprised of an α subunit. 13. The method of Claim 12 wherein the m vi tro efficacy value measured at said o β 3 γ 2 receptor is greater than 20% or the in vi tro efficacy value meaεured εaid α 3 β 3 γ 2 GABA A receptor lε greater than 60%. 14. The method of Claim 13 wherein the in vi tro efficacy value measured at the GABA A receptor comprised of said α 1 subunit is less than 45% or the m vi tro efficacy value measured at the GABA A receptor comprised of said α 5 subunit 15. The method of Claim 12 wherein the m vi tro efficacy value measured at the GABA A receptor comprised of said α subunit is less than 45% or the m vi tro efficacy value measured at the GABA A receptor comprised of said α subunit is less than 40%. 16. The method of Claim 12 wherein the EC 5 „ measured at said α 2 β 3 γ 2 GABA ^ subtype receptor or at said α,β 3 γ 2 GABA. subtype receptor is less than 150 nM. 17. The method of Claim 16 wherein the in vi tro efficacy measured at said α 2 β 3 γ 2 GABA A subtype receptor is greater than 20% or the in vi tro efficacy measured said α 3 β 3 γ 2 GABA A εubtype receptor s greater than 60%. 18. The method of Claim 17 wherein the m vitro efficacy meaεured at the GABA A receptor comprised of said a. subunit is lesε than 45% or the m vi tro efficacy measured at the GABA Λ receptor comprised of said α c subunit is lesε than 40%. 19. The method of Claim 16 wherein the in vi tro efficacy meaεured at the GABA A receptor comprised of said α x subunit is less than 45% or the m vi tro efficacy measured at the GABA A receptor comprised of said α 5 subunit is less than 40%. 20. The method of Claim 12 wherein the GABA A receptor comprised of an α Λ subunit is an a^ 2 y 2 GABA A subtype receptor or the GABA A receptor comprised of an α s subunit is an 5 β 3 γ 2 GABA A subtype receptor. 21. A method for screening a plurality of compoundε εo as to identify at least one compound exhibiting hypnotic activity, comprising: a) selecting a plurality of compounds having a binding affinity of less than 100 nM at any GABA A receptor . b) determining EC 50 and m vi tro efficacy values for each selected compound at an α 2 β 3 γ 2 GABA A subtype receptor or at an α 3 β 3 γ 2 GABA A subtype receptor; c) determining in vitro efficacy values for each selected compound at a GABA A receptor comprised of an ct- L or an α 5 subunit; and d) identifying as exhibiting hypnotic activity each selected compound having an EC b0 value determined in b) of lesε than 200nM, an m vi tro efficacy value measured n b) of greater than 10% for said α 2 β 3 γ 2 GABA A subtype receptor or greater than 50% for said α 3 β 3 γ 2 GABA A subtype receptor, and an in vi tro efficacy value determined m c) of less than 50% for the GABA A receptor comprised of said α subunit or less than 45% for the GABA A receptor comprised of said α 5 subunit. 22. A method for screening a plurality of compounds so as to identify compounds exhibiting hypnotic activity, comprising : a) measuring the EC C0 and in vi tro efficacy of each compound at an α 2 β 3 γ 2 GABA A subtype receptor or an α 3 β 3 γ 2 GABA A subtype receptor; b) measuring the m vi tro efficacy of each compound at a GABA. receptor comprised of an α 2 or α subunit; and c) meaεuring the m vivo effect of each compound m an animal model indicative of hypnotic effects; d) meaεuring the m vivo ef fect of each compound in an animal model indicative of cognitive impairment; and e) identifying a compound aε having hypnotic activity when the EC 50 measured in step a) is less than 200nM, the m vi tro efficacy measured in step a) is greater than 10% for said ct 2 β 3 γ 2 GABA A subtype receptor or greater than 50% for said α 3 β 3 γ 2 GABA A subtype receptor, and the m vi tro efficacy measured m step b) is less than 50% for the GABA A receptor comprised of said a 1 subunit or less than 45% for the GABA A receptor comprised of said α s εubunit and εaid compound produces a statistically significant (p <0.05) positive effect in the animal model indicative of sedation and said compound does not produce a statistically significant effect m the animal model indicative of cognitive impairment . 23. A method for screening a plurality of compounds so as to identify at least one compound exhibiting hypnotic activity, comprising: a) selecting compounds having a binding affinity less than 100 nM at any GABA A receptor; b) measuring the EC„ and m vi tro efficacy of each selected compound at an α 2 β 3 γ 2 GABA A subtype receptor or an ,β 3 γ 2 GABA A subtype receptor; c) measuring the m vi tro efficacy of each selected compound at a GABA A receptor comprised of an c or α 5 subunit; and d) measuring the m vivo effect of each selected compound an animal model indicative of sedative effects ; e) measuring the in vivo effect of each selected compound an animal model indicative of cognitive impairment; and f) identifying as having hypnotic activity each selected compound for which the EC 50 measured m step b) is less than 200nM, the in vi tro eff cacy measured in step b) is greater than 10% for εaid α 2 β 3 γ 2 GABA A εubtype receptor or greater than 50% for εaid α 3 β 3 γ 2 GABA A subtype receptor, and the in vi tro efficacy meaεured εtep c) is lesε than 50% for the GABA A receptor comprised of said α_ subunit or less than 45% for the GABA A receptor comprised of said α s subunit and said compound produces a statistically significant (p <0.05) positive effect m the animal model indicative of hypnotic effects and said compound does not produce a statistically significant effect m the animal model indicative of cognitive impairment. 24. A method for screening a plurality of compounds so as to identify compounds exhibiting anxiolytic activity, comprising : a) determining m vi tro efficacy and EC 50 value for each compound at an α 2 β 3 γ 2 GABA A subtype receptor or an α 3 β 3 γ 2 GABA A subtype receptor; b) determining m vi tro efficacy values for each compound at a GABA A receptor comprised of an α, subunit or an α ς subunit; and c) identifying as exhibiting anxiolytic activity each compound having an EC C0 value determined m a) of less than 200nM and an efficacy value measured m a) greater than the efficacy measured m b) . 25. The method of Claim 24 wherein the EC 5 „ measured m step a) is less than 150 nM. 26. The method of Claim 25 wherein the m vi tro efficacy measured at said 2 β 3 γ 2 or εaid α β 3 γ t GABA A receptor is greater than 20%. 27. The method of Claim 25 wherein the m vi tro ef f icacy measured at said α 2 β 3 γ 2 or εaid α 3 β 3 γ 2 GABA. receptor is greater than 30%. 28. The method of Claim 27 wherein the m vi tro efficacy measured at said GABA A receptor comprised of said α x or said α 5 subunit is less than 20%. 29. The method of Claim 24 wherein the m vi tro efficacy measured at said α 2 β 3 γ 2 or α 3 β 3 γ 2 GABA A receptor is greater than 20%. 30. The method of Claim 24 wherein the m vi tro efficacy measured at said 2 β 3 γ 2 or α 3 β 3 γ 2 GABA A receptor is greater than 30%. 31. The method of Claim 30 wherein the m vi tro ef f icacy measured at εaid GABA A receptor compriεed of said x or said c εubunit is less than 20%. 32. The method of Claim 24 wherein the GABA A receptor comprised of said α subunit is an o^β 2 γ_ GABA A subtype receptor or the GABA receptor comprised of said α s subunit is an a 5 βf{_ . GABA A subtype receptor. 33. A method for screening for compounds having anxiolytic activity, comprising: a) selecting a compound having a binding affinity lesε than 100 nM at any GABA A receptor; b) measuring in vi tro efficacy and EC ζ0 values for each compound at an α 2 β 3 γ 2 or α 3 β 3 γ 2 GABA A receptor; c) measuring m vi tro efficacy values for each compound at a GABA A receptor comprised of an α x or α 5 subunit; and d) selecting a compound having an EC 50 value measured in a) of less than 200nM and an efficacy value measured m b) greater than the efficacy meaεured in c) . 34. A method for screening compounds so as to select at least one compound having anxiolytic activity, comprising-. a) measuring in vi tro efficacy for each compound at an α 2 β 3 2 GABA. subtype receptor or an α 3 β 3 γ 2 GABA A subtype receptor; b) measuring in vitro efficacy and EC C0 values for each compound at a GABA A receptor comprised of an - or α 5 subunit ; c) measuring m vivo effects of each compound in an animal model indicative of anxiolytic activity; d) measuring m vivo effects of each compound in an animal model indicative of sedative effects; and e) selecting each compound having: an EC 50 value meaεured in a) of less than 200nM, an efficacy value measured in b) greater than the efficacy measured in step c) , a statistically significant (p <0.05) positive effect in the animal model indicative of anxiolytic activity, and no statistically significant effect m the animal indicative of sedative effects. 35. A method for screening a plurality of compounds so as to identify at least one compound having anxiolytic activity, comprising: a) selecting a compound having a binding affinity less than 100 nM at any GABA A receptor; b) measuring in vi tro efficacy and EC 50 values for each selected compound at an α 2 β 3 γ 2 or α 3 β 3 γ 2 GABA A receptor; c) measuring m vi tro efficacy for each selected compound at a GABA A receptor comprised of an α x or α 5 subunit; d) measuring in vivo effects of each selected compound in an animal model indicative of anxiolytic activity; e) measuring m vivo effect of each selected compound in an animal model indicative of sedative effects; and f) selecting a compound having: an EC 50 value measured in b) of less than 200nM, an efficacy measured in c) greater than the efficacy measured in d) , a statistically significant (p <0.05) positive effect in the animal model indicative of anxiolytic activity, and no statistically significant effect in the animal indicative of sedative effects. 36. A method for screening a plurality of compounds so as to identify compounds exhibiting antidepressant activity, comprising: a) determining m vi tro efficacy and EC 50 values for each compound using an ct 2 β 3 γ 2 GABA A subtype receptor or an α 3 β 3 γ 2 GABA A subtype receptor; b) determining in vi tro efficacy values for each compound at a GABA A receptor comprised of an a- or an α 5 subunit; and c) identifying as having antidepreεsant activity a compound having an EC 50 value determined in a) of less than 200nM and an efficacy value determined in a) of greater than the efficacy value determined in b) . 37. The method of Claim 36 wherein the EC 50 value determined using said α 2 β 3 γ 2 GABA A subtype receptor or εaid 3 β 3 γ 2 GABA A subtype receptor is less than 150 nM. 38. The method of Claim 37 wherein the in vi tro efficacy measured at said α 2 β 3 γ 2 or said α 3 β 3 γ 2 GABA A receptor is greater than 20%. 39. The method of Claim 37 wherein the in vi tro efficacy measured at said α 2 β 3 γ 2 GABA A subtype receptor or said α 3 β 3 γ 2 GABA A subtype receptor is greater than 30%. 40. The method of Claim 39 wherein the in vi tro efficacy measured at said GABA A receptor compriεed of said α, subunit or said o subunit is lesε than 20% 41. The method of Claim 36 wherein the in vi tro efficacy measured at said α 2 β 3 γ 2 GABA A subtype receptor or said α 3 β 3 γ 2 GABA A subtype receptor is greater than 20%. 42. The method of Claim 36 wherein the in vitro efficacy measured at εaid α 2 β 3 γ 2 GABA A subtype receptor or said α 3 β 3 γ 2 GABA A subtype receptor is greater than 30%. 43. The method of Claim 42 wherein the in vi tro efficacy meaεured at εaid GABA A receptor comprised of said o^ subunit or said α 5 subunit is less than 20%. 44. The method of Claim 36 wherein the GABA A receptor comprised of εaid cq subunit is an a^ 2 y 2 GABA A subtype receptor or the GABA A receptor comprised of said s subunit is an α 5 β 3 γ 2 GABA A subtype receptor. 45. A method for screening compoundε for antidepressant activity, comprising: a) selecting compounds having a binding affinity less than 100 nM at any GABA A receptor; b) determining in vi tro efficacy and EC 50 values for the selected compounds using an α 2 β 3 γ 2 GABA A subtype receptor or an ct 3 β 3 γ 2 GABA A subtype receptor; c) determining in vi tro efficacy for the selected compounds using a GABA A receptor comprised of an λ or an α 5 subunit; and d) identifying as having antidepressant activity a compound having an EC 50 as determined in b) of less than 200nM and an efficacy value as determined in b) greater than the efficacy value determined m c) . 46. A method for screening compounds for antidepressant activity, comprising: a) determining m vitro efficacy and EC 50 values for each compound using an α 2 β 3 γ 2 GABA A subtype receptor or α 3 β 3 γ 2 GABA A subtype receptor; b) determining m vi tro efficacy values for each compound at a GABA A receptor comprised of an a- or an α 5 subunit ; c) determining n vivo effect of said compound m an animal model indicative of antidepressant activity; d) determining the m vivo ef fect of said compound in an animal model indicative of sedative effects; and e) identifying as an antidepressant a compound that produces an EC 50 value as determined in a) of lesε than 200nM, and an efficacy value as determined in b) greater than the efficacy value from c) , and d) produceε a statistically significant (p <0.05) positive effect m the animal model indicative of antidepresεant activity and (11) does not produce a statistically significant effect m the animal model indicative of sedative effects. 47. A method for screening compounds for antidepressant activity, comprising: a) selecting test compounds having a binding affinity lesε than 100 nM at any GABA A receptor; b) determining m vi tro efficacy and EC 50 value for each teεt compound using an α 2 β 3 γ 2 GABA A subtype receptor or an α 3 β 3 γ 2 GABA A subtype receptor; c) determining in vi tro efficacy value for each test compound at a GABA A receptor comprised of an α ^ subunit or an a b subunit ; d) determining the m vivo ef fect of each test compound m an animal model indicative of antidepresεant activity; e) determining the m vivo effect of each test compound in an animal model indicative of sedative effects, and f) identifying as an antidepresεant a compound that produceε. an EC C0 value as determined m b) of less than 200nM, an efficacy value aε determined m c) greater than the efficacy value from d) , and (i) produceε a statistically significant (p <0.05) positive effect in the animal model indicative of antidepressant activity and (11) does not produce a statistically significant effect in the animal model indicative of sedative effects. 48. A method of providing pharmaceutical compounds to patients in need of hypnotic treatment comprising: a) obtaining at least one compound identified as exhibiting hypnotic activity by the method of Claim 21; b) testing said at least one compound and submitting results of said testing as part of submission of information under a United States Federal law which regulates the manufacture, use, or sale of drugs or veterinary products c) showing a pharmaceutical preparation comprising said at least one compound to be εafe for use as required by the provisions of the Federal Food Drug And Cosmetic Act; and d) offering the pharmaceutical preparation for sale m the United States of America for use as an hypnotic drug or hypnotic veterinary product. 49. A method of providing a pharmaceutical preparation to patients in need of anxiolytic treatment comprising: a) obtaining at least one compound identified as exhibiting anxiolytic activity by the method of Claim 24; b) submitting information regarding the anxiolytic activity of εaid at leaεt one compound aε part of an application under a United States Federal law which regulates the manufacture, use, or sale of drugs or veterinary products c) showing a pharmaceutical preparation comprising said at least one compound to be safe for use as required by the provisions of the Federal Food Drug And Cosmetic Act; and d) offering the pharmaceutical preparation for sale in the United States of America for use as an anxiolytic drug or anxiolytic veterinary product. 50. A method of providing a pharmaceutical preparation to patients in need of antidepressant treatment comprising: a) obtaining at leaεt one compound identified as exhibiting antidepressant activity by the method of Claim 36; b) testing said at leaεt one compound and εubmitting results of said testing as part of submission of information under a United States Federal law which regulates the manufacture, use, or sale of drugs or veterinary products c) showing a pharmaceutical preparation comprising εaid at least one compound to be safe for use as required by the provisionε of the Federal Food Drug And Cosmetic Act; and d) offering the pharmaceutical preparation for sale in the United States of America for use as an antidepressant drug or antidepressant veterinary product .

Inventors: ALBAUGH PAMELA, CASSELLA JAMES, CRANDALL MARCI, GALLAGER DOROTHY, RAJACHANDRAN LAVANYA, SHAW KENNETH, TALLMAN JOHN, WHITE GEOFFREY, ALBAUGH, PAMELA, CASSELLA, JAMES, CRANDALL, MARCI, GALLAGER, DOROTHY, RAJACHANDRAN, LAVANYA, SHAW, KENNETH, TALLMAN, JOHN, WHITE, GEOFFREY
Applicants: ALBAUGH PAMELA, CASSELLA JAMES, CRANDALL MARCI, GALLAGER DOROTHY, NEUROGEN CORP, RAJACHANDRAN LAVANYA, SHAW KENNETH, TALLMAN JOHN, WHITE GEOFFREY, ALBAUGH, PAMELA, CASSELLA, JAMES, CRANDALL, MARCI, GALLAGER, DOROTHY, NEUROGEN CORPORATION, RAJACHANDRAN, LAVANYA, SHAW, KENNETH, TALLMAN, JOHN, WHITE, GEOFFREY
Application year: 2000
Main classification: G01N33
IPC codes: A01K67, A61K45, A61P25, A61P43, G01N33