EP1509505A2 - Nicht-endogene, konstitutiv aktivierte rezeptoren für menschliches serotonin und modulatoren von kleiner molekülgrösse - Google Patents

Nicht-endogene, konstitutiv aktivierte rezeptoren für menschliches serotonin und modulatoren von kleiner molekülgrösse

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Publication number
EP1509505A2
EP1509505A2 EP03705889A EP03705889A EP1509505A2 EP 1509505 A2 EP1509505 A2 EP 1509505A2 EP 03705889 A EP03705889 A EP 03705889A EP 03705889 A EP03705889 A EP 03705889A EP 1509505 A2 EP1509505 A2 EP 1509505A2
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EP
European Patent Office
Prior art keywords
phenyl
methyl
bromo
pyrazol
urea
Prior art date
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Pending
Application number
EP03705889A
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English (en)
French (fr)
Inventor
Bradley Teegarden
Keith Drouet
Honnappa Jayakumar
William Thomsen
Paul Maffuid
Katie Elwell
Richard Foster
Michael Lawless
Qian Liu
Julian Smith
Konrad Feichtinger
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Arena Pharmaceuticals Inc
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Arena Pharmaceuticals Inc
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Publication date
Application filed by Arena Pharmaceuticals Inc filed Critical Arena Pharmaceuticals Inc
Publication of EP1509505A2 publication Critical patent/EP1509505A2/de
Pending legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
    • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
    • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D231/14—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
    • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D231/14—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D231/16—Halogen atoms or nitro radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond

Definitions

  • the present invention relates to certain pyrazole derivatives of Formula (I) and pharmaceutical compositions thereof that modulate the activity of the 5-HT 2A serotonin receptor.
  • Compounds and pharmaceutical compositions thereof are directed to methods useful in the prophylaxis or treatment of reducing platelet aggreagation, coronary artery disease, myocardial infarction, transient ischemic attack, angina, stroke, atrial fibrillation, reducing the risk of blood clot formation, asthma or symptoms thereof, agitation or a symptom, behavioral disorders, drug induced psychosis, excitative psychosis, Gilles de la Tourette's syndrome, manic disorder, organic or NOS psychosis, psychotic disorder, psychosis, acute schizophrenia, chronic schizophrenia and NOS schizophrenia and related disorders.
  • the present invention also relates to the method of prophylaxis or treatment of 5-HT 2A serotonin receptor mediated disorders in combination with a dopamine D2 receptor antagonist such as haloperidol, administered separately or together.
  • a dopamine D2 receptor antagonist such as haloperidol
  • G protein-coupled receptors share a common structural motif. All these receptors have seven sequences of between 22 to 24 hydrophobic amino acids that form seven alpha helices, each of which spans the membrane. The transmembrane helices are joined by strands of amino acids having a larger loop between the fourth and fifth transmembrane helix on the extracellular side ofthe membrane. Another larger loop, composed pi ⁇ rnarily of hydrophilic amino acids, joins transmembrane helices five and six on the intracellular side of the membrane. The carboxy tenninus of the receptor lies intracellularly with the amino teiminus in the extracellular space.
  • G protein-coupled receptors exist in the cell membrane in equiUbrium between two different states or conformations: an "inactive" state and an "active" state. As shown schematically in Figure 2, a receptor in an inactive state is unable to link to the intracellular transduction pathway to produce a biological response. Changing the receptor conformation to the active state allows linkage to the transduction pathway and produces a biological response.
  • a receptor may be stabilized in an active state by an endogenous ligand or an exogenous agonist ligand.
  • Recent discoveries such as, including but not exclusively limited to, modifications to the amino acid sequence of the receptor provide means other than ligands to stabilize the active state conformation. These means effectively stabilize the receptor in an active state by simulating the effect of a ligand binding to the receptor. Stabilization by such ligand-independent means is termed "constitutive receptor activation.”
  • Serotouin receptors [0004] Receptors for serotonin (5-hyc_oxyrrypt_mine, 5-HT) are an important class of G protein-coupled receptors. Serotonin is thought to play a role in processes related to learning and memory, sleep, thermoregulation, mood, motor activity, pain, sexual and aggressive behaviors, appetite, neurodegenerative regulation, and biological rhythms. Not surprisingly, serotonin is linked to pathophysiological conditions such as anxiety, depression, obsessive-compulsive disorders, schizophrenia, suicide, autism, migraine, emesis, alcoholism, and neurodegenerative disorders.
  • these types of therapeutics can generally be divided into two classes, the "typical” and the “atypical.” Both have anti- psychotic effects, but the typicals also include concomitant motor-related side effects (extra pyramidal syndromes, e.g., lip-smacking, tongue darting, locomotor movement, etc). Such side effects are thought to be associated with the compounds interacting with other receptors, such as the human dopamine D2 receptor in the nigro-striatal pathway. Therefore, an atypical treatment is preferred. Haloperidol is considered a typical anti-psychotic, and clozapine is considered an atypical anti-psychotic.
  • Serotonin receptors are divided into seven subfamilies, referred to as 5-HT1 through
  • 5-HT7 inclusive. These subfamilies are further divided into subtypes.
  • the 5-HT2 subfamily is divided into three receptor subtypes: 5-HT 2A , 5-HT 2B , and 5-HT 2 c.
  • the human 5-HT 2 c receptor was first isolated and cloned in 1987, and the human 5-HT 2A receptor was first isolated and cloned in 1990. These two receptors are thought to be the site of action of hallucinogenic drugs.
  • antagonists to the 5-HT 2A and 5-HT 2C receptors are believed to be useful in treating depression, anxiety, psychosis, and eating disorders. [0006] U.S.
  • Patent Number 4,985,352 describes the isolation, characterization, and expression of a functional cDNA clone encoding the entire human 5-HT1C receptor (now known as the 5-HT 2C receptor).
  • U.S. Patent Number 5,661,012 describes the isolation, characterization, and expression of a functional cDNA clone encoding the entire human 5-HT 2A receptor.
  • Casey describes a mutation of the cysteine residue at position 322 of the rat 5-HT 2A receptor to lysine (C322K), glutamine (C322Q), and arginine (C322R) which reportedly led to constitutive activation.
  • Herrick-Davis 1 and Herrick-Davis 2 describe mutations of the serine residue at position 312 of the rat 5-HT 2C receptor to phenylalanine (S312F) and lysine (S312K), which reportedly led to constitutive activation.
  • the present invention relates to non-endogenous, constitutively activated forms ofthe human 5-HT 2A and human 5-HT 2C receptors and various uses of such receptors. Further disclosed are small molecule modulators of these receptors. Most preferably, these modulators have inverse agonist characteristics at the receptor.
  • the present invention discloses nucleic acid molecules and the proteins for three non-endogenous, constitutively activated human serotonin receptors, referred to herein as, AP-1, AP-3, and AP-4.
  • the AP-1 receptor is a constitutively active form of the human 5- HT 2 c receptor created by an S31 OK point mutation.
  • the AP-3 receptor is a constitutively active form ofthe human 5-HT2A receptor whereby the intracellular loop 3 (IC3) portion and the cytoplasmic-tail portion of the endogenous human 5-HT2A receptor have been replaced with the IC3 portion and the cytoplasmic-tail portion of the human 5-HT 2C receptor.
  • the AP-4 receptor is a constitutively active form ofthe human 5-HT2A receptor whereby (1) the region ofthe intracellular third loop between the proline of the transmembrane 5 region (TM5) and the proline of TM6 of the endogenous human 5- HT 2A receptor has been replaced with the corresponding region of the human 5-HT 2C receptor (including a S310K point mutation); and (2) the cytoplasmic-tail portion ofthe endogenous human 5- HT 2A receptor has been replaced with the cytoplasmic-tail portion of the endogenous human 5-HT 2C receptor.
  • the invention also provides assays that may be used to directly identify candidate compounds as agonists, partial agonists or inverse agonists to non-endogenous, constitutively activated human serotonin receptors; such candidate compounds can then be utilized in pharmaceutical composition(s) for treatment of diseases and disorders which are related to the human 5-HT 2A and/or human 5-HT 2C receptors.
  • the invention also provides compounds that exhibit high selectivity 5-HT 2A activity.
  • Ri is H, halogens, NR 5 R 6 , OH or OR 7 , wherein
  • R 5 and R_ are independently H, or C 1-6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR_R 9 , NR_R 9 , NHCOCH 3 , OCF 3 , SMe, COOR 10 , S0 3 R 8 , SO_NR_R 9 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 .
  • R 5 and Re may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, OCF 3 , SMe, COOR 10 , S0 2 NR_R 9 , SO 3 R 10 , NHCOCH 3 , COEt, COMe, or halogen;
  • R 8 and R 9 are independently a H, or C ⁇ _ 6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF 3 ,
  • Rio is H or C ⁇ -6 alkyl;
  • R 7 is H or C 1-6 alkyl;
  • R 2 is H, straight chain or branched C 1-6 alkyl, C 2-6 alkenyl, or cycloalkyl;
  • R 3 is halogen, carboxy, CN, alkoxycarbonyl, straight chain or branched C ⁇ _ 6 alkyl, C 2- 6 alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C ⁇ _ 6 alkyl, C 2 .
  • 6 alkenyl, C 2 _ 6 alkynyl, cycloalkyl, aryl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from OH, ORio, NR 8 R 9 , halogen, -C(p) 3j or -0-C(p) 3 where p is halogen, and said cycloalkyl, aryl or heteroaryl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position;
  • R 4 is C ⁇ - 6 alkyl, C 2 . ⁇ alkenyl, or cycloalkyl;
  • Rn and R ⁇ 2 are each independently H, straight chain or branched C ⁇ . 6 alkyl, C 2-6 alkenyl, or cycloalkyl;
  • R- 1 6 and R 17 are each independently H, halogen, CN, NR 8 R 9 , COOR 10 , SR 10 , straight chain or branched C . . . alkyl, C 2 . 6 alkenyl, C 2-6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C ⁇ _ 6 alkyl, C 2 . 6 alkenyl, C 2 .
  • 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , OR 10 , NR_R 9 , halogen, -C(p) 3> or -0-C(p) 3 where p is halogen, and said cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position;
  • L 2 is -0-Q 2 wherein Q 2 is straight chain or branched C ⁇ . 6 alkyl, C 2 . 6 alkenyl, C 2-6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl, wherein each of said cycloalkyl, aryl, alkylaryl, or arylalkyl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , OR 7 , halogen, -C(p) 3 ⁇ or -0-C(p) 3 where p is halogen, or an aliphatic or aromatic heterocycle, and said cycloalkyl, aryl, alkylaryl, and arylalkyl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position; or a pharmaceutically acceptable salt.
  • bold typeface indicates the location of the mutation in the non- endogenous, constitutively activated receptor relative to the corresponding endogenous receptor.
  • Figure 1 shows a generalized structure of a G protein-coupled receptor with the numbers assigned to the transmembrane helices, the intracellular loops, and the extracellular loops.
  • Figure 2 schematically shows the active and inactive states for a typical G protein- coupled receptor and the linkage ofthe active state to the second messenger transduction pathway.
  • Figure 3a provides the nucleic acid sequence of the endogenous human 5-HT 2A receptor (SEQ.TD.NO.:22).
  • Figure 3b provides the corresponding amino acid sequence ofthe endogenous human 5-HT2A receptor (SEQ._D.NO.:23).
  • Figure 4a provides the nucleic acid sequence of the endogenous human 5-HT 2C receptor (SEQ.ID.NO.:24).
  • Figure 4b provides the corresponding amino acid sequence ofthe endogenous human
  • 5-HT 2C receptor SEQ.ID.NO.:25.
  • Figure 5a provides the nucleic acid sequence of a constitutively active form of the human 5-HT 2C receptor ("AP-1 cDNA" - SEQ.ID.NO.:26).
  • Figure 5b provides the corresponding amino acid sequence ofthe AP-1 cDNA ("AP-
  • Figure 6a provides the nucleic acid sequence of a constitutively active form of the human 5-HT 2A receptor whereby the IC3 portion and the cytoplasmic-tail portion of the endogenous 5-HT 2A receptor have been replaced with the IC3 portion and the cytoplasmic-tail portion of the human 5-HT 2C receptor ("AP-3 cDNA" - SEQ.ID.NO.:28).
  • Figure 6b provides the corresponding amino acid sequence ofthe AP-3 cDNA CAP ⁇
  • Figure 6c provides a schematic representation of AP-3, where die dashed-lines represent the portion obtained from the human 5-HT 2C receptor.
  • Figure 7a provides the nucleic acid sequence of a constitutively active form of the human 5-HT 2A receptor whereby (1) the region o the between the proline of TM5 and the proline of TM6 of the endogenous human 5-HT 2A receptor has been replaced with the corresponding region of the human 5-HT 2C receptor (including a S310K point mutation); and (2) the cytoplasmic-tail portion of the endogenous 5-HT 2A receptor has been replaced with the cytoplasmic-tail portion of the endogenous human 5-HT 2C receptor ("AP-4 cDNA" - SEQ.ED.NO.:30).
  • Figure 7b provides the corresponding amino acid sequence ofthe AP-4 cDNA ("AP-
  • Figure 7c provides a schematic representation of the mutated 5-HT 2A receptor of
  • Figure 8 is a representation of the preferred vector, pCMV, used herein.
  • Figure 9 is a diagram illustrating (1) enhanced ( 35 S)GTP ⁇ S binding to membranes prepared from COS cells expressing the endogenous human 5-HT 2C receptor in response to serotonin, and (2) inhibition by mianserin using wheatgerm agglutinin scintillation proximity beads.
  • the concentration of ( 35 S)GTP">S was held constant at 0.3 nM, and the concentration of GDP was held at 1 ⁇ M.
  • the concentration ofthe membrane protein was 12.5 ⁇ g.
  • Figure 10 is a diagram showing serotonin stimulation of ( 35 S)GTP ⁇ S binding to membranes expressing AP-1 receptors in 293T cells and the inhibition by 30 ⁇ M mianserin on WallacTM scintistrips.
  • FIGs 11A and UB are diagrams showing the effects of protein concentration on ( 35 S)GTP-yS binding in membranes prepared from 293T cells transfected with the endogenous human 5-HT 2C receptors and AP-1 receptors compared to cells transfected with the control vector (pCMV) alone in the absence ( Figure 11 A) and presence ( Figure 11B) of 10 ⁇ M serotonin.
  • the radiolableled concentration of ( 35 S)GTP' S was held constant at 0.3 nM, and the GDP concentration was held constant at 1 ⁇ M.
  • the assay was perfonned on 96-well format on WallacTM scintistrips.
  • Figure 12 provides bar-graph comparisons of inositol tris-phosphate ("IP3") production between the endogenous human 5-HT 2A receptor and AP-2, a mutated form of the receptor.
  • IP3 inositol tris-phosphate
  • Figure 13 provides bar-graph comparisons of inositol tris-phosphate ("IP3") production between the endogenous human 5-HT2A receptor and AP-4, a mutated form of the receptor.
  • IP3 inositol tris-phosphate
  • Figure 14 provides bar graph comparisons of IP3 production between the endogenous human 5-HT2A receptor and AP-3, a mutated form ofthe receptor.
  • Figure 15 provides bar-graph comparisons of IP3 production between the endogenous human 5-HT 2C receptor and AP-1.
  • Figures 16A-C provides representative autoradiograms showing displacement of I 125 -
  • Figures 17A-C show in vivo response of animals to Compound 2 exposure.
  • AGONISTS shall mean moieties that activate the intracellular response when they bind to the receptor, or enhance GTP binding to membranes.
  • AMINO ACID ABBREVIATIONS used herein are set out in TABLE 1 :
  • PARTIAL AGONISTS shall mean moieties which activate the intracellular response when they bind to the receptor to a lesser degree/extent than do agonists, or enhance GTP binding to membranes to a lesser degree/extent than do agonists.
  • ANTAGONIST shall mean moieties that competitively bind to the receptor at the same site as the agonists but which do not activate the intracellular response initiated by the active form of the receptor, and can thereby inhibit the intracellular responses by agonists or partial agonists. ANTAGONISTS do not diminish the baseline intracellular response in the absence of an agonist or partial agonist.
  • CANDIDATE COMPOUND shall mean a molecule (for example, and not limitation, a chemical compound) which is amenable to a screening technique.
  • COMPOSITION shall mean a material comprising at least two compounds or two components; for example, and not limitation, a Pharmaceutical Composition is a Composition.
  • COMPOUND EFFICACY shall mean a measurement of the ability of a compound to inhibit or stimulate receptor functionality, as opposed to receptor binding affinity.
  • CONSTITUTIVELY ACTIVATED RECEPTOR shall mean a receptor subject to constitutive receptor activation.
  • CONSTITUTIVE RECEPTOR ACTIVATION shall mean stabilization of a receptor in the active state by means other than binding of the receptor with its endogenous ligand or a chemical equivalent thereof.
  • CONTACT or CONTACTING shall mean bringing at least two moieties together, whether in an in vitro system or an in vivo system.
  • ENDOGENOUS shall mean a material that a mammal naturally produces.
  • ENDOGENOUS in reference to, for example and not limitation, the term "receptor” shall mean that which is naturally produced by a mammal (for example, and not limitation, a human) or a virus.
  • NON-ENDOGENOUS in this context shall mean that which is not naturally produced by a mammal (for example, and not limitation, a human) or a virus.
  • a receptor which is not constitutively active in its endogenous form, but when manipulated becomes constitutively active, is most preferably referred to herein as a "non-endogenous, constitutively activated receptor.”
  • Both terms can be utilized to describe both "in vivo" and "in vitro" systems.
  • the endogenous or non-endogenous receptor may be in reference to an in vitro screening system.
  • screening of a candidate compound by means of an in vivo system is viable.
  • INHIBIT or INHIBITING in relationship to the term “response” shall mean that a response is decreased or prevented in the presence of a compound as opposed to in the absence of the compound.
  • INVERSE AGONISTS shall mean moieties that bind the endogenous form of the receptor or to the constitutively activated form ofthe receptor, and which inhibit the baseline intracellular response initiated by the active form of the receptor below the normal base level of activity which is observed in the absence of agonists or partial agonists, or decrease GTP binding to membranes.
  • the baseline intracellular response is inhibited in the presence ofthe inverse agonist by at least 30%, more preferably by at least 50%, and most preferably by at least 75%, as compared with the baseline response in the absence ofthe inverse agonist.
  • LIGAND shall mean an endogenous, naturally occurring molecule specific for an endogenous, naturally occurring receptor.
  • MODULATE or MODULATING shall mean to refer to an increase or decrease in the amount, quality, response or effect of a particular activity, function or molecule.
  • Compounds which modulate/capable of modulating the 5-HT 2A activity include agonists, inverse agonists, antagonists, inhibitors, activators, and compounds which directly or indirectly affect regulation ofthe 5-HT 2A activity.
  • PHARMACEUTICAL COMPOSITION shall mean a composition comprising at least one active ingredient, whereby the composition is amenable to investigation for a specified, efficacious outcome in a mammal (for example, and not limitation, a human).
  • a mammal for example, and not limitation, a human.
  • STIMULATE or STIMULATING in relationship to the term “response” shall mean that a response is increased in the presence of a compound as opposed to in the absence ofthe compound.
  • the compounds disclosed herein are believed to be useful in the treatment of several additional diseases and disorders, and in the amelioration of symptoms thereof. Without limitation, these include the following:
  • Antiplatelet agents are prescribed for a variety of conditions. For example, in coronary artery disease they are used to help prevent myocardial infarction or stroke in patients who are at risk of developing obstructive blood clots (e.g., coronary thrombosis).
  • a myocardial infarction heart attack
  • the heart muscle does not receive enough oxygen-rich blood as a result of a blockage in the coronary blood vessels. If taken while an attack is in progress or ⁇ nmediately afterward (preferably within 30 minutes), antiplatelets can reduce the damage to the heart.
  • a transient ischemic attack (“TIA” or "mini-stroke”) is a brief imterruption of oxygen flow to the brain due to decreased blood flow through arteries, usually due to an obstructing blood clot.
  • Antiplatelet drugs have been found to be effective in preventing TIAs.
  • Angina is a temporary and often recurring chest pain, pressure or discomfort caused by inadequate oxygen-rich blood flow (ischemia) to some parts of the heart.
  • ischemia oxygen-rich blood flow
  • antiplatelet therapy can reduce the effects of angina and the risk of myocardial infarction.
  • Stroke is an event in which the brain does not receive enough oxygen-rich blood, usually due to blockage of a cerebral blood vessel by a blood clot. In high-risk patients, taking antiplatelets regularly has been found to prevent the formation blood clots that cause first or second strokes.
  • Angioplasty is a catheter based technique used to open arteries obstructed by a blood clot. Whether or not stenting is performed immediately after this procedure to keep the artery open, antiplatelets can reduce the risk of forming additional blood clots following the procedure(s).
  • Coronary bypass surgery is a surgical procedure in which an artery or vein is taken from elsewhere in the body and grafted to a blocked coronary artery, rerouting blood around the blockage and through the newly attached vessel. After the procedure, antiplatelets can reduce the risk of secondary blood clots.
  • Atrial fibrillation is the most common type of sustained irregular heart rhythm
  • Atrial fibrillation affects about two million Americans every year. In atrial fibrillation, the atria (the heart's upper chambers) rapidly fire electrical signals that cause them to quiver rather than contract normally. The result is an abnormally fast and highly irregular heartbeat. When given after an episode of atrial fibrillation, antiplatelets can reduce the risk of blood clots forming in the heart and traveling to the brain (embolism).
  • 5-HT 2A receptors are expressed on smooth muscle of blood vessels and 5-HT secreted by activated platelets causes vasoconstriction as well as activation of additional platelets during clotting.
  • 5-HT 2A inverse agonist will inhibit platelet aggregation and thus be a potential treatment as an antiplatelet therapy. See Satimura, K, et al., Clin Cardiol 2002 Jan. 25 (l):28-32; and Wilson, H.C et al., Thromb Haemost 1991 Sep 2;66(3):355-60.
  • the 5-HT 2A inverse agonists disclosed herein provide beneficial improvement in microcirculation to patients in need of antiplatelet therapy by antagonizing the vasoconstrictive products of the aggregating platelets in, for example and not limitation, the indications described above. Accordingly, in some embodiments, the present invention provides methods for reducing platelet aggregation in a patient in need thereof comprising administering to said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein.
  • the present invention provides methods for treating coronary artery disease, myocardial infarction, transient ischemic attack, angina, stroke, atrial fibrillation, or a symptom of any of the foregoing in a patient in need of said treatment, comprising administering to said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein.
  • the present invention provides methods for reducing risk of blood clot formation in a angioplasty or coronary bypass surgery patient, or a patient suffering from atrial fibrillation, comprising administering to a said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein at a time where such risk exists.
  • the present invention provides methods for treating asthma in a patient in need of said treatment, comprising administering to said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein.
  • methods for treating a symptom of asthma in a patient in need of said treatment comprising administering to said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein.
  • Agitation is a well-recognized behavioral syndrome with a range of symptoms, including hostility, extreme excitement, poor impulse control, tension and uncooperativeness (See Cohen-Mansfield J, and Billig, N., (1986), Agitated Behaviors in the Elderly. I. A Conceptual Review. J Am Geriatr Soc 34(10): 711-721).
  • Alzheimer's disease is a common occurrence in the elderly and often associated with dementia such as those caused by Alzheimer's disease, Lewy Body, Parkinson's, and Huntington's, which are degenerative diseases of the nervous system and by diseases that affect blood vessels, such as stroke, or multi-infarct dementia, which is caused by multiple strokes in the brain can also induce dementia.
  • Alzheimer's disease accounts for approximately 50 to 70% of all dementias (See Koss E, et al., (1997), Assessing patterns of agitation in Alzheimer's disease patients with the Cohen-Mansfield Agitation Inventory. The Alzheimer's Disease Cooperative Study. Alzheimer Dis Assoc Disord ll(suppl 2):S45-S50).
  • Agitated behaviors can also be manifested in cognitively intact elderly people and by those with psychiatric disorders other than dementia
  • the present invention provides methods for treating agitation in a patient in need of such treatment comprising administering to said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein.
  • the agitation is due to a psychiatric disorder other than dementia.
  • the present invention provides methods for treatment of agitation or a symptom thereof in a patient suffering from dementia comprising administering to said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein.
  • the dementia is due to a degenerative disease of the nervous system, for example and without limitation, Alzheimers disease, Lewy Body, Parkinson's disease, and Huntington's disease, or dementia due to diseases that affect blood vessels, including with out limitation stroke and multi-infarct dementia.
  • methods are provided for treating agitation or a symptom thereof in a patient in need of such treatment, where the patient is a cognitively intact elderly patient, comprising administering to said patient a composition comprising a 5-HT 2A inverse agonist disclosed herein.
  • Schizophrenia is a psychopathic disorder of unknown origin, which usually appears for the first time in early adulthood and is marked by a number of characteristics, psychotic symptoms, progression, phasic development and deterioration in social behavior and professional capability in the region below the highest level ever attained.
  • Characteristic psychotic symptoms are disorders of thought content (multiple, fragmentary, incoherent, implausible or simply delusional contents or ideas of doctrine) and of mentality (loss of association, flight of imagination, incoherence up to incomprehensibility), as well as disorders of perceptibility (hallucinations), of emotions (superficial or inadequate emotions), of self-perception, of intentions and impulses, of interhuman relationships, and finally psychomotoric disorders (such as catatonia). Other symptoms are also associated with this disorder. (See, American Statistical and Diagnostic Handbook).
  • Haloperidol is a potent dopamine D2 receptor antagonist. It is widely prescribed for acute schizophrenic symptoms, and is very effective for the positive symptoms of schizophrenia. However, Haldol is not effective for the negative symptoms of schizophrenia and may actually induce negative symptoms as well as cognitive dysfunction. In accordance with some methods of the invention, adding a 5-HT 2A inverse agonist concomitantly with Haldol will provide benefits including the ability to use a lower dose of Haldol without losing its effects on positive symptoms, while reducing or eliminating its inductive effects on negative symptoms, and prolonging relapse to the patient's next schizophrenic event.
  • Haloperidol is used for treatment of a variety of behavioral disorders, drug induced psychosis, excitative psychosis, Gilles de la Tourette's syndrome, manic disorders, psychosis (organic and NOS), psychotic disorder, psychosis, schizophrenia (acute, chronic and NOS). Further uses include in the treatment of infantile autism, huntington's chorea, and nausea and vomiting from chemotherapy and chemotherapeutic antibodies. Administration of 5-HT 2A inverse agonists disclosed herein with haloperidol also will provide benefits in these indications.
  • the present invention provides methods for treating a behavioral disorder, drug induced psychosis, excitative psychosis, Gilles de la Tourette's syndrome, manic disorders, psychosis (organic and NOS), psychotic disorder, psychosis, schizophrenia (acute, chronic and NOS) comprising administering to said patient a dopamine D2 receptor antagonist and a 5-HT 2A inverse agonist disclosed herein.
  • the present invention provides methods for treating a behavioral disorder, drug induced psychosis, excitative psychosis, Gilles de la Tourette's syndrome, manic disorders, psychosis (organic and NOS), psychotic disorder, psychosis, schizophrenia (acute, chronic and NOS) comprising administering to said patient haloperidol and a 5-HT 2A inverse agonist disclosed herein.
  • the present invention provides methods for treating infantile autism, huntington's chorea, or nausea and vomiting from chemotherapy or chemotherapeutic antibodies comprising administering to said patient a dopamine D2 receptor antagonist and a 5-HT 2A inverse agonist disclosed herein.
  • the present invention provides methods for treating infantile autism, huntington's chorea, or nausea and vomiting from chemotherapy or chemotherapeutic antibodies comprising administering to said patient haloperidol and a 5-HT 2A inverse agonist disclosed herein.
  • the present invention provides methods for treating schizophrenia in a patient in need of said treatment comprising administering to said patient a dopamine D2 receptor antagonist and a 5-HT 2A inverse agonist disclosed herein.
  • the dopamine D2 receptor antagonist is haloperidol.
  • the administration of the dopamine D2 receptor antagonist can be concomitant with administration ofthe 5-HT 2A inverse agonist, or they can be administered at different times. Those of skill in the art will easily be able to determine appropriate dosing regimes for the most efficacious reduction or elimination of deleterions haloperidol effects.
  • haloperidol and the 5-HT 2A inverse agonist are administered in a single dosage form, and in other embodiments, they are administered in separate dosage forms.
  • the present invention further provides methods of alleviating negative symptoms of schizophrenia induced by the administration of haloperidol to a patient suffering from said schizophrenia, comprising administering to said patient a 5-HT 2A inverse agonist as disclosed herein.
  • sequence information regarding the non-endogenous, constitutively active human 5-HT 2A and 5-HT 2 c receptors are referred to by identifiers as set forth in TABLE 2:
  • AP-2 a mutation analogous to that reported by Casey (C322K) was utilized in the human 5-HT 2A receptor and is referred to herein as AP-2.
  • AP-2 did not lead to sufficient constitutive activation to allow for utilization in screening techniques.
  • Herrick-Davis 1 or Herrick-Davis 2 is of limited predictive value relative to the manipulation ofthe human 5-HT 2 c receptor. Consequently, the ability to make reasonable predictions about the effects of mutations to the rat 5-HT receptors vis-a- vis the corresponding human receptors is not possible. Nonetheless, this unfortunate lack of reasonable predictability provides the opportunity for others to discover mutations to the human 5-HT receptors that provide evidence of constitutive activation.
  • the present invention is based upon the desire of defining mutated sequences ofthe human serotonin receptors 5-HT 2A and 5-HT 2C whereby such mutated versions ofthe expressed receptor are constitutively active. These constitutively active receptors allow for, inter alia, screening candidate compounds.
  • G protein receptor When a G protein receptor becomes constitutively active, it binds to a G protein (Gq, Gs,
  • candidate compounds are identified using the "generic" G protein-coupled receptor assay (i.e. an assay to select compounds that are agonists, partial agonists, or inverse agonists), further screening to confirm that the compounds have interacted at the receptor site is preferred.
  • a compound identified by the "generic” assay may not bind to the receptor, but may instead merely "uncouple" the G protein from the intracellular domain.
  • Lysergic acid diethylamide is a well-known agonist ofthe 5-HT 2A and 5-HT 2C receptors, while mesulergine is a well-known antagonist to the 5-HT 2C receptor. Accordingly, in most preferred embodiments, an agonist (LSD) and/or antagonist (mesulergine) competitive binding assay(s) is used to further screen those compounds selected from the "generic" assay for confirmation of serotonin receptor binding.
  • IP3 accumulation can be used to confirm constitutive activation of these types of Gq coupled receptors (see Herrick-Davis-1).
  • IP3 accumulation assays can be used to further screen those compounds selected from an agonist and/or antagonist competitive binding assay. .
  • Candidate compounds selected for further development can be formulated into pharmaceutical compositions using techniques well known to those in the art. Suitable pharmaceutically- acceptable carriers are available to those in the art; for example, see Remington's Pharmaceutical Sciences, 16 th Edition, 1980, Mack Publishing Co., (Oslo et al., eds.).
  • PCR was performed using either TaqPlusTM precision polymerase (Stratagene) or rTthTM polymerase (Perkin Elmer) with the buffer systems provided by the manufacturers, 0.25 ⁇ M of each primer, and 0.2 mM of each ofthe four (4) nucleotides.
  • the cycle condition was 30 cycles of 94°C for 1 minute, 57 °C for 1 minute and 72 °C for 2 minutes.
  • the 1.5 kb PCR fragment was digested with Xho I and Xba I and subcloned into the Sal I-Xba I site of pBluescript.
  • the cDNA containing a S310K mutation (AP-1 cDNA) in the third intracellular loop , of the human 5-HT 2C receptor was constructed by replacing the Sty I restriction fragment containing amino acid 310 with synthetic double stranded oligonucleotides encoding the desired mutation.
  • the sense strand sequence utilized had the following sequence:
  • the cDNA encoding endogenous human 5-HT 2A receptor was obtained by RT-PCR using human brain poly-A + RNA; a 5 ' primer from the 5 ' untranslated region with a Xho I restriction site:
  • the resulting cDNA clones were fully sequenced and found to encode two amino acid changes from the published sequences.
  • the first change is a T25N mutation in the N-terminal extracellular domain and the second change is an H452Y mutation.
  • These mutations are likely to represent sequence polymorphisms rather than PCR errors since the cDNA clones having the same two mutations were derived from two independent PCR procedures using Taq polymerase from two different commercial sources (TaqPlusTM Stratagene and rTthTM Perkin Elmer).
  • the cDNA containing the point mutation C322K in the third intracellular loop was constructed by using the Sph I restriction enzyme site, which encompasses amino acid 322.
  • a primer containing the C322K mutation: 5'-CAAAGAAAGTACTGGGCATCGTCTTCTTCCT-3' (SEQ.ro.NO.:7) was used along with the primer from the 3' untranslated region set forth above as SEQ.LD.NO.:6.
  • the resulting PCR fragment was then used to replace the 3' end ofthe wild type 5-HT 2A cDNA by the T4 polymerase blunted Sph I site.
  • PCR was performed using pfu polymerase (Stratagene) with the buffer system provided by the manufacturer and 10% DMSO, 0.25 mM of each primer, 0.5mM of each of the 4 nucleotides.
  • the cycle conditions were 25 cycles of 94°C for 1 minute, 60°C for 1 minute, and 72°C for 1 minute.
  • the human 5-HT 2A cDNA with intracellular loop 3 (IC3) or IC3 and cytoplasmic tail replaced by the corresponding human 5-HT 2C cDNA was constructed using PCR-based mutagenesis.
  • (a) Replacement of IC3 Loop [0104] The IC3 loop of human 5-HT 2A cDNA was first replaced with the corresponding human 5-HT 2C cDNA. Two separate PCR procedures were performed to generate the two fragments, Fragment A and Fragment B, that fuse the 5-HT 2C IC3 loop to the transmembrane 6 (TM6) of 5-HT 2A .
  • the 237 bp PCR fragment, Fragment A, containing 5-HT 2C IC3 and the initial 13 bp of 5-HT 2A TM6 was amplified by using the following primers:
  • Second round PCR was performed using Fragment A and Fragment B as co-templates with SEQ.ID.NO.:8 and SEQ.ID.NO.:ll (it is noted that the sequences for SEQ.ID.NOS.:6 and 11 are the same) as primers.
  • the resulting 740 bp PCR fragment, Fragment C contained the IC3 loop of human 5-HT 2C fused to TM6 through the end of the cytoplasmic tail of human 5-HT 2A .
  • PCR was performed using pfuTM polymerase (Stratagene) with the buffer system provided by the manufacturer, and 10% DMSO, 0.25 mM of each primer, and 0.5 mM of each ofthe four (4) nucleotides.
  • the cycle conditions were 25 cycles of 94 °C for 1 minute, 57 °C (1st round PCR) or 60 °C (2nd round PCR) for 1 minute, and 72 °C for 1 minute (1st round PCR) or 90 seconds (2nd round PCR). [0107] To generate a PCR fragment containing a fusion junction between the human 5-HT 2A
  • TM5 and the IC3 loop of 5-HT 2C four (4) primers were used.
  • the two external primers, derived from human 5-HT 2A had the following sequences: 5'-CGTGTCTCTCCTTACTTCA-3' (SEQ.ID.NO.:12)
  • the first internal primer utilized was an antisense strand containing the initial 13 bp of IC3 of 5-HT 2C followed by the terminal 23 bp derived from TM5 of 5-HT 2A : 5'-TCGGCGCAGTACTTTGATAGTTAGAAAGTAGGTGAT-3' (SEQ.ID.NO.:13)
  • the second internal primer was a sense strand containing the te ⁇ ninal 14 bp derived from TM5 of 5-HT 2A followed by the initial 24 bp derived from IC3 of 5-HT 2C :
  • PCR was performed using endogenous human 5-HT 2A and a co-template, Fragment
  • PCR was performed using a sense primer containing the C-te ⁇ ninal 22 bp of TM7 of endogenous human 5-HT 2A followed by the initial 21 bp ofthe cytoplasmic tail of endogenous human 5-HT 2C : 5'-TTCAGCAGTCAACCCACTAGTCTATACTCTGTTCAACAAAATT-3 ' (SEQ.LO.NO.:15)
  • the antisense primer was derived from the 3 ' untranslated region of endogenous human 5-HT 2C : 5'-ATTTCTAGACATATGTAGCTTGTACCGT-3' (SEQ.ID.NO.:16).
  • the resulting PCR fragment, Fragment D contained the last 22 bp of endogenous human 5-HT 2A TM7 fused to the cytoplasmic tail of endogenous human 5-HT 2C .
  • Second round PCR was performed using Fragment D and the co-template was endogenous human 5-HT 2A that was previously digested with Ace I to avoid undesired amplification.
  • the antisense primer used was SEQ.ID.NO.:16 (the sequences for SEQ.LD.NOS.:16 and 2 are the same) and the sense primer used was derived from endogenous human 5-HT 2A : 5 '-ATCACCTACTTTCTAACTA-3 ' (SEQ.ID.NO.:17).
  • PCR conditions were as set forth in Example lB3(a) for the first round PCR, except that the annealing temperature was 48 °C and the extension time was 90 seconds.
  • the resulting 710 bp PCR product was digested with Apa I and Xba I and used to replace the corresponding Apa I-Xba I fragment of either (a) endogenous human 5-HT 2A , or (b) 5-HT 2A with 2C IC3 to generate (a) endogenous human 5-HT 2A with endogenous human 5-HT 2 c cytoplasmic tail and (b) AP-3, respectively.
  • This mutant was created by replacement ofthe region of endogenous human 5-HT 2A from amino acid 247, the middle of TM5 right after Pro 246 , to amino acid 337, the middle of TM6 just before Pro 338 , by the corresponding region of AP-1 cDNA.
  • the junction in TM5 is referred to as the "2 A-2C junction”
  • the junction in TM6 is referred to as the "2C-2A junction.”
  • PCR fragments containing the desired hybrid junctions were generated.
  • the 5' fragment of 561 bp containing the 2A-2C junction in TM5 was generated by PCR using endogenous human 5-HT 2A as template, SEQ.EO.NO.:12 as the sense primer, and the antisense primer was derived from 13 bp of 5-HT 2C followed by 20 bp of 5-HT 2A sequence: 5'-CCATAATCGTCAGGGGAATGAAAAATGACACAA-3' (SEQ.ED.NO.:18)
  • the middle fragment of the 323 bp contains endogenous human 5-HT 2C sequence derived from the middle of TM5 to the middle of TM6, flanked by 13 bp of 5-HT 2A sequences from the 2A-2C junction and the 2C-2A junction.
  • This middle fragment was generated by using AP-1 cDNA as a template, a sense primer containing 13 bp of 5-HT2A followed by 20 bp of 5-HT 2C sequences across the 2A-2C junction and having the sequence:
  • the 3' fragment of 487 bp containing the 2C-2A junction was generated by PCR using endogenous human 5-HT 2A as a template and a sense primer having the following sequence from the 2C-2A junction: 5'-GATCATGTGGTGCCCTTTCTTCATCACAAACAT-3' (SEQ.ED.NO.:21) and the antisense primer was SEQ.ID.NO.:6 (see note above regarding SEQ.ID.NOS.:6 and 11).
  • the M3' PCR used the middle and M3' PCR fragment described above as the co- template, SEQ.ID.NO.:19 as the sense primer and SEQ.ED.NO.:6 (see note above regarding SEQ.ID.NOS.:6 and 11) as the antisense primer, and generated a 784 bp amplification product.
  • the final round of PCR was performed using the 857 bp and 784 bp fragments from the second round PCR as the co-template, and SEQ.ID.NO.:12 and SEQ.ID.NO.:6 (see note above regarding SEQ._O.NOS.:6 and 11) as the sense and the antisense primer, respectively.
  • the 1.32 kb amplification product from the final round of PCR was digested with Pst I and Eco RI. Then resulting 1 kb Pst I-Eco RI fragment was used to replace the corresponding fragment ofthe endogenous human 5-HT 2A to generate mutant 5-HT 2A with 5-HT 2C : C310K/IC3.
  • the Apa I-Xba fragment of AP3 was used to replace the corresponding fragment in mutant 5-HT 2A with 5-HT 2C : C310K/IC3 to generate AP4.
  • the vector utilized be pCMV.
  • This vector was deposited with the American Type Culture Collection (ATCC) on October 13, 1998 (10801 University Boulevard., Manassas, VA 20110-2209 USA) under the provisions ofthe Budapest Treaty for the International Recognition ofthe Deposit of Microorganisms for the Purpose of Patent Procedure. The DNA was tested by the ATCC and determined to be viable. The ATCC has assigned the following deposit number to pCMV: ATCC #203351. See Figure 8.
  • tube A was prepared by mixing 20 ⁇ g DNA (e.g., pCMV vector; pCMV vector AP-1 cDNA, etc.) in 1.2 ml serum free DMEM (Irvine Scientific, Irvine, CA); tube B was prepared by rnixing 120 ⁇ l hpofectarnine (Gibco BRL) in 1.2 ml serum free DMEM. Tubes A and B were then admixed by inversions (several times), followed by incubation at room temperature for 30-45 min. The admixture is referred to as the "transfection mixture".
  • Plated COS-7 cells were washed with IX PBS, followed by addition of 10 ml serum free DMEM. 2.4 ml of the transfection mixture was then added to the cells, followed by incubation for 4 hrs at 37°C/5% C0 2 . The transfection mixture was then removed by aspiration, followed by the addition of 25 ml of DMEM/10% Fetal Bovine Serum. Cells were then incubated at 37°C/5% C0 2 . After 72 hr incubation, cells were then harvested and utilized for analysis.
  • (a) ( 35 S)GTP ⁇ S binding is generically applicable to all G protein-coupled receptors; and (b) ( 35 S)GTP ⁇ S binding is proximal at the membrane surface, thereby making it less likely to pick-up molecules which affect the intracellular cascade.
  • the assay utilizes the ability of G protein-coupled receptors to stimulate ( 35 S)GTP ⁇ S binding to membranes expressing the relevant receptors. Therefore, the assay may be used to directly screen compounds at the disclosed serotonin receptors.
  • Figure 9 demonstrates the utility of a scintillation proximity assay to monitor the binding of ( 35 S)GTP ⁇ S to membranes expressing, e.g., the endogenous human 5-HT 2 c receptor expressed in COS cells.
  • a preferred protocol for the assay is such that the assay was incubated in 20 mM HEPES, pH 7.4, binding buffer with 0.3 nM ( 35 S)GTP ⁇ S and 12.5 ⁇ g membrane protein and 1 ⁇ M GDP for 30 minutes.
  • Wheatgerm agglutinin beads 25 ⁇ l; Amersham
  • the tubes were then centrifuged at 1500 x g for 5 minutes at room temperature and then counted in a scintillation counter.
  • serotonin which as the endogenous ligand activates the 5-HT 2C receptor, stimulated ( 35 S)GTP ⁇ S binding to the membranes in a concentration dependant manner.
  • the stimulated binding was completely inhibited by 30 ⁇ M mianserin, a compound considered as a classical 5-HT 2C antagonist, but also known as a 5-HT 2C inverse agonist.
  • this assay measures agonist-induced binding of ( 35 S)GTP ⁇ S to membranes and can be routinely used to measure constitutive activity of receptors
  • the present cost of wheatgerm agglutinin beads may be prohibitive.
  • a less costly but equally applicable alternative also meets the needs of large-scale screening.
  • Flash plates and WallacTM scintistrips may be used to format a high throughput ( 35 S)GTP ⁇ S binding assay. This technique allows one to monitor the tritiated ligand binding to the receptor while simultaneously monitoring the efficacy via ( 35 S)GTP ⁇ S binding. This is possible because the WallacTM beta counter can switch energy windows to analyze both tritium and 35 S-labeled probes.
  • this assay may be used for detecting of other types of membrane activation events that result in receptor activation.
  • the assay may be used to monitor 32 P phosphorylation of a variety of receptors (including G protein-coupled and tyrosine kinase receptors).
  • the bound ( 35 S)GTP ⁇ S or the 32 P- phosphorylated receptor will activate the scintillant coated on the wells.
  • Use of Scinti ® strips (WallacTM) demonstrate this principle.
  • this assay may be used for measuring ligand binding to receptors using radiolabeled ligands. In a similar manner, the radiolabeled bound ligand is centrifuged to the bottom of the well and activates the scintillant.
  • the ( 35 S)GTP ⁇ S assay results parallel the results obtained in traditional second messenger assays of receptors.
  • Figure 11A and Figure 11B demonstrate the enhanced binding of ( 35 S)GTP ⁇ S to membranes prepared from 293T cells expressing the control vector alone, the native human 5-HT 2 c receptor or the AP-1 receptor was observed (data not shown).
  • the total protein concentration used in the assay affects the total amount of ( 35 S)GTP ⁇ S binding for each receptor.
  • the c.p.m. differential between the CMV transfected and the constitutively active mutant receptor increased from approximately 1000 c.p.m at 10 ⁇ g/well to approximately 6-8000 c.p.m. at 75 ⁇ g/well protein concentration, as shown in Figure 11.
  • the AP-1 receptor showed the highest level of constitutive activation followed by the wild type receptor, which also showed enhanced ( 35 S)GTP ⁇ S binding above basal. This is consistent with the ability of the endogenous human 5-HT 2C receptor to accumulate intracellular IP3 in the absence of 5-HT stimulation (Example 5) and is also consistent with published data claiming that the endogenous human 5-HT 2C receptor has a high natural basal activity. Therefore, the AP-1 receptor demonstrates that constitutive activity may be measured by proximal ( 3S S)GTP ⁇ S binding events at the membrane interface.
  • Membranes were prepared from transfected COS-7 cells (see Example 2) by homogenization in 20 mM HEPES and 10 mM EDTA, pH 7.4 and centrifuged at 49,000 x g for 15 min. The pellet was resuspended in 20 mM HEPES and 0.1 mM EDTA, pH 7.4, homogenized for 10 sec. using a Polytron homogenizer (Brinkman) at 5000 rpm and centrifuged at 49,000 x g for 15 min. The final pellet was resuspended in 20 mM HEPES and 10 mM MgCl 2 , pH 7.4, homogenized for 10 sec. using polytron homogenizer (Brinkman) at 5000 rpm.
  • COS-7 and 293 Cells [0132] On day one, COS-7 cells or 293 cells were plated onto 24 well plates, usually lxl 0 5 cells/well or 2xl0 5 cells/well, respectively. On day two, the cells were transfected by first mixing 0.25 ug DNA (see Example 2) in 50 ⁇ l serum-free DMEM/well and then 2 ⁇ l lipofectamine in 50 ⁇ l serum-free DMEM/well.
  • transfection media The solutions (“transfection media”) were gentiy mixed and incubated for 15-30 minutes at room temperature. The cells were washed with 0.5 ml PBS and then 400 ⁇ l of serum free media was mixed with the transfection media and added to the cells. The cells were then incubated for 3-4 hours at 37°C/5%C0 2 . Then the transfection media was removed and replaced with 1 ml/well of regular growth media. On day 3, the media was removed and the cells were washed with 0.5 ml PBS.
  • tube A was prepared by mixing 20 ⁇ g DNA (e.g., pCMV vector; pCMV vector AP-1 cDNA, etc.) in 1.2 ml serum free DMEM (Irvine Scientific, Irvine, CA); tube B was prepared by mixing 120 ⁇ l lipofectamine (Gibco BRL) in 1.2 ml serum free DMEM. Tubes A and B were then admixed by inversions (several times), followed by incubation at room temperature for 30-45 min.
  • DNA e.g., pCMV vector; pCMV vector AP-1 cDNA, etc.
  • tube B was prepared by mixing 120 ⁇ l lipofectamine (Gibco BRL) in 1.2 ml serum free DMEM. Tubes A and B were then admixed by inversions (several times), followed by incubation at room temperature for 30-45 min.
  • the admixture is referred to as the "transfection mixture”.
  • Plated 293 cells were washed with 1XPBS, followed by addition of 10ml serum free DMEM.
  • 2.4 ml of the transfection mixture was then added to die cells, followed by incubation for 4 hrs at 37°C/5% C0 2 .
  • On day 3 cells were trypsinized and counted, followed by plating of lxlO 6 cells/well (poly D-lysine treated 12-well plates). Cells were permitted to adhere to the wells, followed by one wash with lxPBS. Thereafter, 0.5 ⁇ Ci 3 H-inositol in 1ml inositol-free DMEM was added per well. Protocol B.
  • the cells were washed with 0.5 ml PBS and then 0.45 ml of assay medium was added containing inositol-free/serum free media, 10 ⁇ M pargyline, 10 mM lithium chloride, or 0.4 ml of assay medium and 50 ⁇ l of lOx ketanserin (ket) to a final concentration of 10 ⁇ M.
  • the cells were then incubated for 30 minutes at 37° C. Then the cells were washed with 0.5 ml PBS and 200 ⁇ l of fresh/ice cold stop solution (IM KOH; 18 mM Na-borate; 3.8 mM EDTA) was added/well.
  • IM KOH fresh/ice cold stop solution
  • the solution was kept on ice for 5-10 minutes or until the cells were lysed and then neutralized by 200 ⁇ l of fresh/ice cold neutralization sol. (7.5 % HCL).
  • the lysate was then transferred into 1.5 ml rnicro- centrifuge tubes and 1 ml of chloroform/methanol (1:2) was added/tube.
  • the solution was vortexed for 15 seconds and the upper phase was applied to a Biorad AG1-X8 anion exchange resin ( 100-200 mesh). The resin was washed with water and 0.9 ml of the upper phase was loaded onto the column.
  • the column was washed with 10 ml of 5 mM myo-inositol and 10 ml of 5 mM Na-borate/60mM Na- formate.
  • the inositol trisphosphates were eluted into scintillation vials containing 10 ml of scintillation cocktail with 2 ml of 0.1 M formic acid 1 M ammonium formate.
  • the columns were regenerated by washing with 10 ml of 0.1 M formic acid/3M ammonium formate and rinsed twice with dd H 2 0 and stored at room temperature in water. Results are discussed below.
  • Figure 12 is an illustration of IP3 production from the human 5-HT 2A receptor which was mutated using the same point mutation as set forth in Casey, which rendered the rat receptor constitutively active.
  • the results represented in Figure 12 support die position that when the point mutation shown to activate the rat receptor is introduced into the human receptor, little activation of the receptor is obtained that would allow for appropriate screening of candidate compounds, with the response being only moderately above that of the endogenous human 5-HT 2A receptor. Generally, a response of at least 2X above that ofthe endogenous response is preferred.
  • Figure 13 provides an illustration comparing IP3 production from endogenous 5-HT 2A receptor and the AP4 mutation.
  • the results illustrated in Figure 13 support the position that when the novel mutation disclosed herein is utilized, a robust response of constitutive IP3 accumulation is obtained (e.g., over 2X that ofthe endogenous receptor).
  • Figure 14 provides an illustration of IP3 production from AP3.
  • the results illustrated in Figure 14 support the position fliat when the novel mutation disclosed herein is utilized, a robust response of constitutive IP3 accumulation is obtained.
  • Figure 15 provides bar-graph comparisons of IP3 accumulation between endogenous human 5-HT 2C receptor and AP-1. Note that the endogenous receptor has a high degree of natural constitutive activity relative to the control CMV transfected cells (i.e., the endogenous receptor appears to be constitutively activated).
  • transiently expressing constitutively active mutant human 5-HT 2C receptor AP-1 were incubated with binding buffer (20 mM HEPES, pH 7.4, 100 mM NaCl, 20 mM MgCl 2 ° 6H,0, 0.2% saponin, and 0.2 mM ascobate), GDP(1 ⁇ M) and compound in a 96-well plate format for a period of 60 rninutes at ambient room temperature. Plates were then centrifuged at 4,000 rpm for 15 minutes followed by aspiration of the reaction mixture and counting for 1 minute in a WallacTM MicroBeta plate scintillation counter.
  • Example 7 SCREENING OF SELECTED COMPOUNDS TO CONFIRM RECEPTOR BINDING: AP-1 [0141]
  • the candidate compounds identified from Example 7 were then screened using the assay protocol of Example 4 (mesulergine), using the AP-1 mutant receptor.
  • IC 50 (nM) values were determined; five of the nearly 120 compounds of Example 7 were determined to have potent binding affinity for the receptor. Results are summarized in TABLE 4. TABLE 4
  • the "primary" screen designed to directly identify human 5-HT 2A /5-HT 2C receptor inverse agonists consisted of a membrane-based GTP ⁇ S binding assay utilizing membranes prepared from COS7 cells transiently transfected with AP-1 human receptor.
  • Candidate compounds (lO ⁇ M final assay concentration) directly identified as inhibiting receptor-mediated increases in GTP ⁇ S binding by greater than 50-75% (arbitrary cut-off value) were considered active "hits”.
  • Primary assay hits were then re-tested in the same assay to recorifirm their inverse agonist activity.
  • This protocol is substantially the same as set forth above in Example 6.
  • Primary screening assays measuring GTP ⁇ S binding to membranes prepared from COS7 cells transiently transfected with human mutated 5-HT 2C receptor (AP-1) were used to directly identify inverse agonists in screening libraries (Tripos, Inc.).
  • Candidate compound screens were performed in a total assay volume of 200 ⁇ l using scintillant-coated Wallac ScintistripTM plates.
  • the primary assay was comprised of the following chemicals (at indicated final assay concentrations): 20 mM HEPES, pH 7.4, 100 mM NaCl, 20 mM MgCl 2 , 0.2% saponin, 0.2 mM ascorbic acid, 1 ⁇ M GDP, 0.3 nM GTP ⁇ 35 S, and 12.5 ⁇ g of the above defined membranes. Incubations were performed for 60 minutes at ambient room temperature. The binding assay incubation was temiinated by centrifugation of assay plates at 4,000 rpm for 15 minutes, followed by rapid aspiration ofthe reaction mixture and counting in a Wallac MicroBetaTM scintillation counter.
  • Radioligand binding competition experiments were performed in a total assay volume of 200 ⁇ l using standard 96-well microtiter plates.
  • the final assay ingredients consisted of assay buffer (20 mM HEPES and 10 mM MgCl 2 ), InM ( 3 H) mesulergine, and 50 ⁇ g of membranes (COS7 with AP-1 as defined above).
  • Nonspecific ( 3 H) mesulergine binding was defined in the presence of 100 ⁇ M mianserin. Incubations were performed for 1 hour at 37°C.
  • Receptor bound radioligand was resolved from free radioligand by rapid filtration ofthe assay mixture over a Wallac FiltermatTM Type B filter, followed by washing with ice-cold assay buffer using a SkatronTM cell harvester. Radioactivity was counted using a Wallac 1205 BetaPlateTM counter. Each assay plate contained five negative control wells (membranes expressing receptor and no candidate compound addition) and three positive control wells (each containing 100 ⁇ M mianserin). For one concentration tests, candidate compounds were diluted into assay buffer and screened at a final concentration of 10 ⁇ M, in triplicate. For IC 50 determinations, candidate compounds were diluted in assay buffer and eight different concentrations were evaluated, in triplicate. A total of 16 wells were designated for an eight concentration mianserin dose response curve evaluation for both assays.
  • Radioligand binding competition experiments were performed in a total assay volume of 200 ⁇ l using standard 96-well microtiter plates.
  • the final assay ingredients comprised assay buffer (20 mM HEPES and lOmM MgCl 2 ), InM ( 3 H)LSD, and 50 ⁇ g of the above-defined membranes (COS7 with AP-1).
  • Nonspecific ( 3 H)LSD binding was defined in the presence of 100 ⁇ M serotonin. Incubations were performed for 1 hour at 37° C.
  • Receptor bound radioligand was resolved from free radioligand by rapid filtration of the assay mixture over a Wallac FiltermatTM Type B filter, followed by washing with ice-cold assay buffer using a SkatronTM cell harvester. Radioactivity was counted using a Wallac 1205 BetaPlateTM counter. Each assay plate contained five negative control wells (membranes expressing receptor and no candidate compound addition) and three positive control wells (containing 100 ⁇ M mianserin). For one concentration tests, candidate compounds were diluted into assay buffer and screened at a final concentration of 10 ⁇ M in triplicate. For IC 50 determinations, candidate compounds were diluted in assay buffer and eight different concentrations were evaluated in triplicate. A total of 16 wells were designated for an eight concentration serotonin dose response curve evaluation for both assays. EXAMPLE 9D
  • Candidate compound identified in the assays of Examples 9A-9C were then evaluated for inositol phosphate accumulation, following the protocol of Example 5 (COS7 cells expressing human mutated 5-HT 2A receptor, AP-3), modified as follows: tube A was prepared by mixing 16 ⁇ g DNA (e.g., pCMV vector; pCMV vector AP-1 cDNA, etc.) in 1.0 ml serum free DMEM (Irvine Scientific, Irvine, CA); tube B was prepared by mixing 60 ⁇ l lipofectamine (Gibco BRL) in 1.0 ml serum free DMEM.
  • DNA e.g., pCMV vector; pCMV vector AP-1 cDNA, etc.
  • tube B was prepared by mixing 60 ⁇ l lipofectamine (Gibco BRL) in 1.0 ml serum free DMEM.
  • Tubes A and B were then admixed by inversions (several times), followed by incubation at room temperature for 30 min. The admixture is referred to as die "transfection mixture”. Plated 293 cells were washed with 10 ml Serum Free DMEM, followed by addition of 11 ml Serum Free DMEM. 2.0 ml ofthe transfection mixture was then added to the cells, followed by incubation for 5 hrs at 37° C/5% C0 2 . On day 3, cells were trypsinized and counted, followed by plating of lxlO 6 cells/well (12-well plates). Cells were permitted to adhere to the wells for 8 hrs, followed by one wash with lxPBS. Thereafter, 0.5 ⁇ Ci 3 H-inositol in 1ml inositol-free DMEM was added per well.
  • the cells were washed with 1.5 ml PBS and then 0.9 ml of assay medium was added containing inositol-free/serum free media, 10 ⁇ M pargyline, 10 mM lithium chloride, for 5min in 37°C/5% C0 2 followed by lOO ⁇ l addition of candidate compound diluted in the same material. The cells were then incubated for 120 minutes at 37° C. Then the cells were washed witii 1.5 ml PBS and 200 ⁇ l of fresh/icecold stop solution (IM KOH; 18 mM Na-borate; 3.8 mM EDTA) was added/well.
  • IM KOH fresh/icecold stop solution
  • the solution was kept on ice for 5-10 minutes or until the cells were lysed and then neutralized by 200 ⁇ l of fresh/ice cold neutralization sol. (7.5 % HCL).
  • the lysate was then transferred into 1.5 ml micro-centrifuge tubes and 1 ml of chloroform/methanol (1:2) was added/tube.
  • the solution was vortexed for 15 seconds and the upper phase was applied to a Biorad AG1-X8 anion exchange resin (100-200 mesh).
  • the resin was washed with water and 0.9 ml ofthe upper phase was loaded onto the column.
  • the column was washed with 10 ml of 5 mM myo-inositol and 10 ml of 5 mM Na-borate/60mM Na-formate.
  • the inositol trisphosphates were eluted into scintillation vials containing 10 ml of scintillation cocktail with 2 ml of 0.1 M formic acid/ 1 M ammonium formate.
  • the columns were regenerated by washing with 10 ml of 0.1 M formic acid/3M ammonium formate and rinsed twice with ddH 2 0 and stored at room temperature in water.
  • This series of compounds exhibits highly selective 5-HT 2A activity. Accordingly, in one aspect of die invention, a series of compounds possessing 5-HT 2A receptor activity that are useful as inverse agonists at such receptors is designated by the general Formula (I):
  • Ri is H, halogens, NR 5 R_, OH or OR 7 , wherein
  • R5 and Re are independently H, or C ⁇ _ 6 alkyl, or C 2-6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONRgR . , NRsR . , NHCOCH 3 , OCF 3 , SMe, COOR 10 , S0 3 R 8 , S0 2 NR 8 R 9 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 .
  • R 5 and R ⁇ may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, OCF 3 , SMe, COOR 10 , S0 2 NR_R 9 , SO 3 R ⁇ 0 , NHCOCH 3 , COEt, COMe, or halogen;
  • R 8 and R 9 are independently a H, or C ⁇ _ 6 alkyl, or C 2 _ 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF 3 , OEt, CC1 3 ,
  • R 7 is H or C ⁇ alkyl; ii) R 2 is H, straight chain or branched C ⁇ __ alkyl, C 2-6 alkenyl, or cycloalkyl; iii) R 3 is halogen, carboxy, CN, alkoxycarbonyl, straight chain or branched C ⁇ -6 alkyl, C 2- ⁇ alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C_._ alkyl, C 2 . 6 alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from OH, OR 10 , NRsRg, halogen,
  • R 4 is Cue alkyl, C 2-6 alkenyl, or cycloalkyl;
  • Rn and R ⁇ 2 are each independently H, straight chain or branched C ⁇ - 6 alkyl, C 2-6 alkenyl, or cycloalkyl;
  • R ⁇ 3 , R 14 , Ri5, Ri ⁇ and R are each independently H, halogen, CN,
  • NR 8 R 9 COORio, SRio, straight chain or branched C ⁇ -6 alkyl, C_. 6 alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C ⁇ _ 6 alkyl, C 2 . 6 alkenyl, C 2 .
  • 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , ORio, NR_R 9 , halogen, -C(p) 3 ⁇ or -0-C(p) 3 where p is halogen, and said cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can alternatively or additionally be optionally substituted witii up to four alkyl substituents in any position;
  • L_ is -O-Q 2 wherein Q 2 is straight chain or branched C ⁇ -6 alkyl, C 2 _ 6 alkenyl, C 2 _ 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl, wherein each of said cycloalkyl, aryl, alkylaryl, or arylalkyl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , OR .
  • An aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to 4 hetero atoms independently selected from N, O, or S) or a 6 membered aromatic non-heterocyclic ring or a polycycle.
  • C ⁇ -6 alkyl moieties can be straight chain or branched; optionally substituted C ⁇ _ 6 alkyl moieties can be straight chain or branched; C 2 . 6 alkenyl moieties can be straight chain or branched; and optionally substituted C 2 . 6 alkenyl moieties can be straight chain or branched.
  • suitable C ⁇ -6 alkyl groups include but are not limited to methyl, ethyl, n-propyl, i-propyl, n-butyl, and t-butyl.
  • Halogens are typically F, CI, Br, and I.
  • Examples of 5 or 6 membered ring moieties include, but are not restricted to, phenyl, furanyl, thienyl, imidazolyl, pyridyl, pyrrolyl, oxazolyl, isoxazolyl, triazolyl, pyrazolyl, tetrazolyl, thiazolyl and isothiazoyl.
  • Examples of polycycle moieties include, but are not restricted to, naphthyl, benzothiazolyl, benzofuranyl, benzimidazolyl, quinolyl, isoquinolyl, indolyl, quinoxalinyl, quinazolinyl and benzothienyl.
  • B is L b q is 1, m is 0, and n is 0.
  • B is L q is 1, m is 1, and n is 0.
  • B is L b q is 1, m is 0, and n is 1.
  • B is L q is 0, m is 0, and n is 0.
  • B is L 2 .
  • Ri is H, CI, F, dimethylamino, pyrrolidin-1-yl, morpholin-1-yl, 4-methylpiperazin-l-yl, OH or OCH 3 .
  • R 2 is H and R 4 is methyl of each of the foregoing embodiments of the compounds of Formula (I),
  • R ⁇ 3 , R 14 , R 15 , Ri 6 and Rn are each independently H, F, CI, Br, CN, dimethylamino, ethoxycarbonyl, methylthio, methyl, ethyl, isopropyl, trifluoromethyl, trifluoromethoxy, methoxy, NH 2 or N0 2 .
  • R 2 is H and R4 is methyl.
  • B is L 2 .
  • R 4 is methyl.
  • R 2 is H.
  • R 2 is H or lower allcyl(C ⁇ _ ⁇ );
  • R 3 is lower alkyl (C ⁇ _ 6 ), or halogen
  • R 4 is lower alkyl ( . 6 );
  • X is eitiier Oxygen or Sulfur
  • R 15 is H or lower alkyl(C ⁇ _();
  • Ri 6 and Rn are independently d . 6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, C0NR I8 R 19 ,
  • NR 1S R 19 NHCOCH 3j OCF 3 , SMe, COOR 20 , SO 3 R 20 , S0 2 NR 18 R 19 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 . 6 alkenyl, H, halogens, C M alkoxy, C 3-6 cycloalkyl, C ⁇ . ⁇ alkyl, and aryl;
  • Ris and R ⁇ 9 are independently a H or C ⁇ -6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 21 R 22 , NR 21 R 22 , NHCOCH 3 , OCF 3 , SMe, C00R 23 , S0 3 R 23 , S0 2 NR 2 ⁇ R 22 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 .
  • R !8 and R ⁇ 9 may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, OCF 3 , SMe, COOR 23 , S0 2 NR 21 R 22 , S0 3 R 23 , NHCOCH 3 , COEt, COMe, or halogen;
  • R 20 and R 23 are each independently selected from H or C ⁇ _ 6 alkyl;
  • R 2 ⁇ and R 22 are each independently are independently a H, or C) .
  • an aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to 4 hetero atoms independently selected from N, 0, or S) or a 6 membered aromatic non-heterocyclic ring or a polycycle;
  • Ci- ⁇ alkyl groups can be straight chain or branched; optionally substituted C ⁇ . 6 alkyl moieties can be straight chain or branched; C 2 . 6 alkenyl moieties can be straight chain or branched; and optionally substituted C 2 .6 alkenyl moieties can be straight chain or branched.
  • suitable Ci- ⁇ alkyl groups include but are not limited to methyl, ethyl, n- propyl, i-propyl, n-butyl, and t-butyl.
  • Halogens are typically F, CI, Br, and I.
  • Examples of 5 or 6 membered ring moieties include, but are not restricted to, phenyl, furanyl, thienyl, imidazolyl, pyridyl, pyrrolyl, oxazolyl, isoxazolyl, friazolyl, pyrazolyl, tefrazolyl, thiazolyl and isothiazoyl.
  • Examples of polycycle moieties include, but are not restricted to, naphthyl, benzothiazolyl, benzofuranyl, benzimidazolyl, quinolyl, isoquinolyl, indolyl, quinoxalinyl, quinazolinyl and benzothienyl.
  • R 3 is F, CI, Br, I, C ⁇ -6 straight chain or branched alkyl, C 3 . 8 cycloalkyl, C 4-9 alkylcycloalkyl, or C 2 . 6 alkenyl;
  • X is O or S
  • Y is NR 15 Ri6, or (CH 2 ) m R 17 , or 0(CH 2 ) n R ; m is an integer between 0 and 4, inclusive; n is an integer between 0 and 4, inclusive;
  • R 4 is H, C ⁇ - 8 straight chain or branched alkyl, C 3-8 cycloalkyl, C 4-9 alkylcycloalkyl, or C 2 . 8 alkenyl;
  • R 2 and R ⁇ 5 a is each independently selected from H, C ⁇ _ 8 straight chain or branched alkyl, C 3 . 8 cycloalkyl, C 4-9 alkylcycloalkyl, or C 2-8 alkenyl;
  • Ri 6 and R 17 is each independently selected from: C_._ straight chain or branched alkyl, C 2-8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl, or CH 2 aryl, wherein each moiety within said C ⁇ -8 straight chain or branched alkyl, C 2 .
  • alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl or CH 2 aryl may be optionally substituted by up to four substituents in any position, whereby each substituent is independently selected from: H, F, CI, Br, I, R 20 , CF 3 , CF 2 R 7 , CF 2 CF 2 , CC1 3 , CC1 2 R 7 , CC1 2 CC1 2 R 7 , NR 18 R 19 , NR ⁇ 8 COR 20 , NR 18 SO 2 R 20 , OR 20 , OCF 3 , OCF 2 R 20 , OCF 2 CF 2 R 20 , OCOR 20 , OSO 2 R 20 , OPO(OR 20 ) 2 , SR 20 , SCF 3 , SCF 2 R 20 , SCF 2 CF 2 R 20 , SCOR 20 , SO 3 R 20 , S0 2 NR 1S R 19 , PO(OR 20 ) 3 , PO(
  • R 20 is H, Ci-s straight chain or branched alkyl, C 3-8 cycloalkyl, C 4-9 alkylcycloalkyl, C 2-8 alkenyl, aryl or alkylaryl; R ⁇ 8 and R ⁇ 9 are each independently selected from H, C ⁇ -8 straight chain or branched alkyl, C 2 .
  • An aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to
  • C . .. alkyl groups include but are not limited to methyl, ethyl, n-propyl, i-propyl, n-butyl, and t-butyl.
  • Examples of 5 or 6 membered ring moieties include, but are not restricted to, phenyl, furanyl, thienyl, imidazolyl, pyridyl, pyrrolyl, oxazolyl, isoxazolyl, friazolyl, pyrazolyl, tefrazolyl, thiazolyl, and isothiazolyl.
  • Examples of polycycle moieties include, but are not restricted to, naphthyl, benzothiazolyl, benzofuranyl, benzirnidazolyl, quinolyl, isoquinolyl, indolyl, quinoxalinyl, quinazolinyl, and benzothienyl.
  • X is O or S;
  • R 4 is H or CH 3 ;
  • Rs and R 20 are each independently selected from H, C ⁇ -8 straight chain or branched alkyl, C 3 . 8 cycloalkyl, C 4-9 alkylcycloalkyl, or C 2-8 alkenyl;
  • R 15 is H, F, CI, Br, I, R 20 , CF 3 , CF 2 R 20 , CF 2 CF 2 , CC1 3 , CC1 2 R 20 , CC1 2 CC1 2 R 20 , NR 18 R 19 ,
  • R 18 and R !9 are each independently selected from H, C ⁇ -8 straight chain or branched alkyl, C 2 . 8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl, or CH 2 aryl, wherein each moiety within said C ⁇ _ 8 straight chain or branched alkyl, C 2-8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl or CH 2 aryl may be optionally substituted by up to four substituents in any position, whereby each substituent is independently selected from: F, CI, Br, I, CF 3 , CC1 3) CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , NH 2 , NHCH 3 , N(CH 3 ) 2 , NHC 2 H 5 , N(C 2 H 5 ) 2 , NHC 3 H 7 , N(C 3 H 7 ) 2 , NHC 4 H 9 ,
  • R 13 , Ri 4 , R 15 , Ri 6 and R 17 each independently selected from the following: F, CI, Br, I, CF 3 , CC1 3 , CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , NH 2 , NHCH 3; N(CH 3 ) 2 , NHC 2 H 5 , N(C 2 H 5 ) 2 , NHC 3 H 7 , N(C 3 H 7 ) 2 , NHC 4 H 9 , N(C 4 H 9 ) 2 , NHCOH, NHCOCH 3 , NHCOC 2 H 5 , NHCOC 3 H 7 , NHCOC 4 H 9 , NHS0 2 CH 3 , NHS0 2 C 2 H 5 , NHS0 2 C 3 H 7 , NHS0 2 C 4 H 9 , OH, OCH 3 , OC 2 H 5 , OC 3 H 7 , OC 4 H 7 , OC 4 H 9 , OC 5 H 9 , OC 5
  • a more preferred series of compounds possessing 5-HT 2A receptor activity that are useful as inverse agonists at such receptors is designated by the general Formula (B):
  • R 2 is H or lower alkyl (C 1-4 );
  • R 3 is Me, or Et, or halogen
  • X is either Oxygen or Sulfur
  • Y is NR 1 5R16, or (CH 2 ) m R 17 , or 0(CH 2 ) n R 17 ;
  • Ri 5 is H or lower alkyl(C ); Ri 6 and Rn are independently C ⁇ _ 6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 18 R ⁇ 9 , NR ⁇ 8 R 19 , NHCOCH 3 , OCF 3 , SMe, COOR 20 , SO 3 R 20 , S0 2 NR ⁇ 8 R 19 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2-6 alkenyl, H, halogens, C__ ⁇ alkoxy, C 3-6 cycloalkyl, C ⁇ -6 alkyl, aryl and aryloxy wherein each of the C 1-4 alkoxy, C 3 .6 cycloalkyl
  • Ris and R ⁇ 9 are independently a H or C ⁇ _ 6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 21 R 22 , NR 21 R 22 , NHCOCH3, OCF 3 , SMe, COOR 23 , S0 3 R 23 , S0 2 NR_ ⁇ R 22 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 . 6 alkenyl, H, halogens, Q.
  • C 3-6 cycloalkyl, C ⁇ -6 alkyl, and aryl or R ⁇ 8 and R ⁇ 9 may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 ,
  • R 20 and R 23 are each independently selected from H or C ⁇ . 6 alkyl;
  • R 21 and R 22 are each independently are independently a H, or C 1 . 6 alkyl, or C 2- 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF 3 , OEt, CC1 3 , Me, N0 2 , OH, OMe, SMe, COMe, CN, COOR 20 , SO 3 R 20 , COEt, NHCOCH 3 , or aryl;
  • C 1 . 6 alkyl moieties can be straight chain or branched; optionally substituted C ⁇ _ 6 alkyl moieties can be straight chain or branched:
  • C 2-s alkenyl moieties can be straight chain or branched; and optionally substituted C 2 _6 alkenyl moieties can be straight chain or branched.
  • C ⁇ _ 6 alkyl groups include but are not limited to methyl, ethyl, n- propyl, i-propyl, n-butyl, and t-butyl.
  • Halogens are typically F, Cl, Br, and I.
  • Examples of 5 or 6 membered ring moieties include, but are not restricted to, phenyl, furanyl, thienyl, imidazolyl, pyridyl, pyrrolyl, oxazolyl, isoxazolyl, friazolyl, pyrazolyl, tefrazolyl, thiazolyl and isothiazoyl.
  • Examples of polycycle moieties include, but are not restricted to, naphthyl, benzothiazolyl, benzofuranyl, benzimidazolyl, quinolyl, isoquinolyl, indolyl, quinoxahnyl, quinazolinyl and benzothienyl.
  • R 2 and R ⁇ 5 are H;
  • R 3 is Br;
  • X is O;
  • R 4 is Me.
  • R ⁇ 6 is preferably 4-trifluoromethoxyphenyl , 4- trifluoromethoxybenzyl, 4-chlorophenyl or 4-fluorophenyl.
  • Certain preferred compounds are: Compound 7 N-(3-(4-bromo-2-methylpyrazol-3-yl)phenyl)(((4-frifluoromethoxy)phenyl)an nocarboxamide
  • Inositol phosphate accumulation assays evidence the activity of test compounds. Both single concentration percentages of control values and IC 0 determinations indicate activity.
  • the column legends have the following meanings: IP 3 % Control: The values in this column reflect an IP Accumulation Assay where the test compounds were evaluated at one concentration of 10 ⁇ M. For these assays, the compound was diluted into inositol-free Dulbecco's Eagle Media containing 10 ⁇ M pargyline and 10 mM LiCl and tested at a final assay concentration of 10 ⁇ M, in triplicate. The percent control value was calculated based on the control in which no test compound was added.
  • IPT AP-3 ICsn nM The values in this column reflect an IP accumulation assay in which the test compound was evaluated at several different concentrations whereby an IC 50 could be detennined. This column corresponds to the column appearing in the tables above which is labeled: Inositol Phosphate Accumulation, AP-3, IC 50 Value ( ⁇ M).
  • WT 5-HTTA LSD ICsn nM The values in this column reflect a competitive binding assay using LSD. This column corresponds to the column appearing in the tables above which is labeled: Competitive Binding, WT 5-HT 2A) (( 3 H)LSD), IC 50 Value ( ⁇ M).
  • R 2 is H.
  • R 3 is Br.
  • X is O.
  • R 4 is Me.
  • R 17 is 4-methoxyphenyl or tertiary butyl.
  • 5-HT 2A receptor was further confirmed by the following:
  • Lysergic acid diethylamide is a potent 5-HT 2A receptor and dopamine D2 receptor ligand.
  • An indication of the selectivity of compounds for either or both of these receptors involves displacement of radiolabeled-bound LSD from pre-treated brain sections.
  • radiolabeled I 125 -LSD NN Life Sciences, Boston, Mass., Catalogue number NEX-199 was utilized; spiperone (RBI, Natick, Mass. Catalogue number s-128) a 5-HT 2A receptor and dopamine D2 receptor antagonist, was also utilized.
  • Buffer consisted of 50 nanomolar TRIS-HCl, pH 7.4.
  • Brain sections were incubated in (a) Buffer plus 1 nanomolar I i25 -LSD; (b) Buffer plus 1 nanomolar I 125 -LSD and 1 micromolar spiperone; or Buffer plus 1 nanomolar I 125 -LSD and 1 micromolar Compound 1 for 30 minutes at room temperature. Sections were then washed 2x 10 minutes at 4 ° C. in Buffer, followed by 20 seconds in distilled H 2 0. Slides were then air-dried.
  • Figures 16A-C provide representative autoradiographic sections from this study.
  • Figure 16A evidences darker bands (derived from I 125 -LSD binding) primarily in both die fourth layer of the cerebral cortex (primarily 5-HT 2A receptors), and the caudate nucleus (primarily dopamine D2 receptors and some 5-HT 2A receptors).
  • spiperone which is a 5-HT 2 and dopamine D2 antagonist, displaces the I 125 -LSD from these receptors on both the cortex and the caudate.
  • Compound 1 appears to selectively displace the I 125 - LSD from the cortex (5-HT 2A ) and not the caudate (dopamine D2).
  • a third series of compounds having 5-HT 2A receptor activity is represented by a class
  • R 2 is H.
  • R 3 is Br.
  • X is O.
  • R 4 is Me.
  • R is preferably 4-trifluoromethoxyphenyl, or thiophene, or
  • a 5-HT 2A receptor antagonist or inverse agonist is expected to decrease amphetamine- stimulated locomotion without affecting baseline locomotion. See, for example, Soresnon, et al, 266(2) J. Pharmacol. Exp. Ther. 684 (1993). Based upon the foregoing information, Compound 2 is a potent inverse agonist at the human 5-HT 2A receptor. For the following study, the following parameters and protocol were utilized:
  • a San Diego Instruments Flex Field apparatus was used to quantify baseline and amphetamine-stimulated locomotor activity. This apparatus consists of four 16" x 16" clear plastic open fields. Photocell arrays (16 in each dimension) interfaced with a personal computer to automatically quantify activity. Several measures of activity can be assessed witii the apparatus, including total photocell beam breaks. Animals (vehicle control and Compound treated) were injected s.c. 30 minutes prior to initiation of analysis. Following this 30 minute period, animals were placed individually into an open field and baseline activity was assessed for 30 minutes (habituation phase).
  • R 2 is H or lower alkyl(C );
  • R 3 is Me, or Et, or halogen;
  • X is either Oxygen or Sulfur;
  • Y is NR 15 R 16 , or (CH2) m Rn, or 0(CH 2 ) n R 17 ;
  • R 15 is H or lower alkyl(C M );
  • R 18 and R ⁇ 9 are independently a H or C ⁇ -6 alkyl, or C 2-6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 21 R 22 , NR 2 ⁇ R 22 , NHCOCH 3 , OCF 3 , SMe, C00R 23 , S0 3 R 23 , S0 2 NR 2 ⁇ R 22 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2-5 alkenyl, H, halogens, C..
  • Ris and R 19 may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, OCF 3 , SMe, COOR 7 , S0 2 NR 2 ⁇ R 22 , S0 3 R 23 , NHCOCH 3 , COEt, COMe, or halogen;
  • Rn is C ⁇ - 6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 21 R 22 , NR_ ⁇ R 22 , NHCOCH,, OCF 3 , SMe, COOR 23 , S0 3 R 23 , S0 2 NR 21 R 22 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 .
  • R 20 and R 23 may be independently selected from H or C ⁇ -6 alkyl;
  • R 21 and R 22 are independently a H, or C alkyl, or C 2-6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF3, OEt, CC1 3 , Me, N0 2 , OH, OMe, SMe, COMe, CN, COOR 20 , SO 3 R 20 , COEt, NHCOCH 3 , or aryl.
  • An aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to 4 hetero atoms independently selected from N, O, or S) or a 6 membered aromatic non-heterocyclic ring or a polycycle;
  • C ⁇ -6 alkyl moieties can be straight chain or branched; optionally substituted C ⁇ _ 6 alkyl moieties can be straight chain or branched; C 2-6 alkenyl moieties can be straight chain or branched; and optionally substituted C 2 . 6 alkenyl moieties can be straight chain or branched;
  • suitable C ⁇ -6 alkyl groups include but are not limited to methyl, ethyl, n-propyl, i-propyl, n-butyl, and t-butyl.
  • Halogens are typically F, CI, Br, and I.
  • Examples of 5 or 6 membered ring moieties include, but are not restricted to, phenyl, furanyl, thienyl, imidazolyl, pyridyl, pyrrolyl, oxazolyl, isoxazolyl, friazolyl, pyrazolyl, tefrazolyl, thiazolyl and isothiazolyl.
  • Examples of polycycle moieties include, but are not restricted to, naphthyl, benzothiazolyl, benzofuranyl, benzimidazolyl, quinolyl, isoquinolyl, indolyl, quinoxahnyl, quinazolinyl and benzothienyl.
  • Some embodiments of the invention are compounds of Formula (V) and have one of the structures below.
  • X is O or S; P is H or CH 3 ;
  • R 8 and R 30 are each independently selected from H, C ⁇ -8 straight chain or branched alkyl, C 3-8 cycloalkyl, C 4-9 alkylcycloalkyl, or C 2 . 8 alkenyl;
  • R 15 is H, F, CI, Br, I, R 20 , CF 3 , CC1 3 , NR 18 R 19 , NR 30 COR 20 , NR 30 SO 2 R 20 , OR 20 , OCF 3 ,
  • R ⁇ S and said adjacent position can together be selected from SCH 2 S, SCH 2 CH 2 S, OCH 2 0, or OCH 2 CH 2 0 to form a bi-cyclic structure;
  • R 20 is H, C ⁇ -8 straight chain or branched alkyl, C 3 . 8 cycloalkyl, C 4-9 alkylcycloalkyl, C 2-8 alkenyl, aryl or alkylaryl;
  • Ris and Ri are each independently selected from: H, C ⁇ _ 8 straight chain or branched alkyl, C 2- 8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl or CH 2 aryl wherein each moiety within said C ⁇ -8 straight chain or branched alkyl, C 2 .
  • alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl or CH 2 aryl may be optionally substituted by up to four substituents in any position, whereby each substituent is independently selected from: F, CI, Br, I, CF 3 , CC1 3 , CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , NH 2 , NHCH 3 , N(CH 3 ) 2 , NHC 2 H 5 , N(C 2 H 5 ) 2 , NHC 3 H 7 , N(C 3 H 7 ) 2 , NHC 4 H 9 , N(C 4 H 9 ) 2 , NHCOH, NHCOCH 3 , NHCOC 2 H 5 , NHCOC 3 H 7 , NHCOC 4 H 9 , NHS0 2 CH 3 , NHS0 2 C 2 H 5 , NHS0 2 C 3 H 7 , NHS0 2 C 4 H 9 , OH, OCH 3 , OC
  • Ar is a phenyl ring optionally substituted with up to five groups selected from tiie group consisting of halogen, OR 7 , OH, NR_R 9 , carboxy, CN, alkoxycarbonyl, straight chain or branched C ⁇ -6 alkyl -C(p) 3 ⁇ or -0-C(p) 3 where p is halogen;
  • R 8 and R 9 are independently a H, or C ⁇ -6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF 3 ,
  • R 8 and R 9 may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 ,
  • R 7 is H or C 1-6 alkyl
  • Rio is H or C ⁇ -6 alkyl
  • R 2 is H, straight chain or branched C ⁇ -6 alkyl, C 2 . 6 alkenyl, or cycloalkyl;
  • R 3 is halogen, carboxy, CN, alkoxycarbonyl, straight chain or branched C M alkyl, C 2 _ 6 alkenyl, C 2 solicit6 alkynyl, cycloalkyl, aryl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched -6 alkyl, C 2 . 6 alkenyl, C 2 .
  • 6 alkynyl, cycloalkyl, aryl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from OH, ORio, NR 8 R 9 , halogen, -C(p) 3> or -O- C(p) 3 where p is halogen, and said cycloalkyl, aryl or heteroaryl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position;
  • R is Ci -6 alkyl, C 2 . 6 alkenyl, or cycloalkyl;
  • Rn and R l2 are each independently H, straight chain or branched C ⁇ _ 6 alkyl, C 2-6 alkenyl, or cycloalkyl;
  • Ri 3 , R ⁇ , R 1 5, Ri ⁇ and R are each independently H, halogen, CN, NR 8 R 9 , COORio, SRio, straight chain or branched C 1-6 alkyl, C 2 . 6 alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C ⁇ _ 6 alkyl, C 2 .
  • 6 alkenyl, C 2-6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , OR 10 , NR 8 R 9 , halogen, -C(p) 3 , or -0-C(p) 3 where p is halogen, and said cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position;
  • L 2 is -0-Q 2 wherein Q 2 is straight chain or branched - ⁇ alkyl
  • the present invention further provides methods for modulating die activity of a human 5-HT 2A serotonin receptor by contacting the receptor with a compound of formula:
  • Ri is H, halogens, NR 5 R_, OH or OR 7 , wherein
  • R 5 and Re are independently H, or C ⁇ -6 alkyl, or C 2-6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 8 R 9 , NR 8 R 9 , NHCOCH 3 , OCF 3 , SMe, COOR 10 , S0 3 R 8 ,
  • 6 cycloalkyl, Q .6 alkyl, or aryl groups may be further optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 8 R 9 , NR 8 R 9 , NHCOCH 3 , OCF 3 , SMe, COOR 10 , S0 2 NR 8 R 9 , SO 3 R 10 , COMe,
  • COEt CO-lower alkyl, SCF 3 , CN, C 2 . 6 alkenyl, H, halogens, C M alkoxy, C 3 . 6 cycloalkyl, C 1-6 alkyl, and aryl; or
  • R 5 and ⁇ may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, OCF 3 , SMe, COOR 10 , S0 2 NRsR 9 , SO 3 R ⁇ 0 , NHCOCH 3 , COEt, COMe, or halogen; R 8 and R 9 are independently a H, or C .
  • Rio is H or C M alkyl;
  • R 7 is H or C ⁇ _6 alkyl;
  • R 2 is H, straight chain or branched C ⁇ -6 alkyl, C 2 . 6 alkenyl, or cycloalkyl;
  • R 3 is halogen, carboxy, CN, alkoxycarbonyl, straight chain or branched Ci . alkyl, C 2- 6 alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C e alkyl, C 2 .
  • 6 alkenyl, C 2 _6 alkynyl, cycloalkyl, aryl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from OH, OR 10 , NR ⁇ R 9 , halogen,
  • R 4 is C ⁇ _6 alkyl, C 2-6 alkenyl, or cycloalkyl;
  • L is:
  • R u and R ⁇ 2 are each independently H, straight chain or branched C M alkyl, C 2-6 alkenyl, or cycloalkyl;
  • R 13 , R 14 , R 15 , R 16 and R 17 are each independently H, halogen, CN,
  • NR 8 R 9 , COOR JO , SR 10 straight chain or branched C ⁇ -6 alkyl, C 2 . 6 alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C ⁇ _ 6 alkyl, C 2 . 6 alkenyl, C 2 .
  • 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , ORio, NR 8 R 9 , halogen, -C(p) 3j or -0-C(p) 3 where p is halogen, and said cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position;
  • Q 2 is straight chain or branched _ 6 alkyl
  • An aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to
  • Also provided by the present invention are methods for modulating the activity of a human 5-HT 2A serotonin receptor by contacting the receptor with a compound of formula:
  • R 2 is H or lower alkyl(C ⁇ _ ⁇ );
  • R 3 is lower alkyl (C ⁇ . 6 ), or halogen;
  • R 4 is lower alkyl (C ⁇ _ 6 );
  • X is eidier Oxygen or Sulfur
  • R 1 6 and Rn are independently C ⁇ _ 6 alkyl, or C 2 . ⁇ alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 18 R ⁇ 9 , NR 18 R 19 , NHCOCH 3 , OCF 3 , SMe, COOR 20 , SO 3 R 20 , S0 2 NR 18 R 19 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 . 6 alkenyl, H, halogens, C alkoxy, C 3-6 cycloalkyl, d_6 alkyl, and aryl;
  • Ris and R 19 are independently a H or C ⁇ -6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, C0NR 21 R 22 , NR 21 R 22 , NHCOCH 3 , OCF 3 , SMe, C00R 23 , S0 3 R 23 , S0 2 NR 2 ⁇ R 22 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2-6 alkenyl, H, halogens, .
  • Ris and R ⁇ 9 may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 ,
  • R 20 and R 23 are each independently selected from H or C ⁇ _ 6 alkyl;
  • R 2 ⁇ and R 22 are each independently are independently a H, or C 1-6 alkyl, or C 2- 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF 3 , OEt, CC1 3 , Me, N0 2 , OH, OMe, SMe, COMe, CN, COOR 20 , SO 3 R 20 , COEt, NHCOCH 3 , or aryl; an aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to 4 hetero atoms independently selected from N, O, or S) or a 6 membered aromatic non-heterocyclic ring or a polycycle.
  • the present invention further provides methods for modulating the activity of a human 5-HT 2A serotonin receptor by contacting the receptor with a compound of formula:
  • R 3 is F, Cl, Br, I, C 1-6 straight chain or branched alkyl, C 3-8 cycloalkyl, C 4 . 9 alkylcycloalkyl, or C 2 . 6 alkenyl; Xis O or S;
  • Y is NR15R16, or (CH 2 ) m R 17 , or 0(CH 2 ) n R 17; m is an integer between 0 and 4, inclusive; n is an integer between 0 and 4, inclusive; is H, C ⁇ -8 straight chain or branched alkyl, C 3-8 cycloalkyl, C 4-9 alkylcycloalkyl, or C 2-8 alkenyl;
  • R 2 and R 15 a is each independently selected from H, C ⁇ . g straight chain or branched alkyl, C 3 . 8 cycloalkyl, C 4-9 alkylcycloalkyl, or C 2-8 alkenyl;
  • R J6 and Rn is each independently selected from: C ⁇ . 8 straight chain or branched alkyl, C 2-8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl, or CH 2 aryl, wherein each moiety within said C 1-8 straight chain or branched alkyl, C 2 .
  • alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl or CH 2 aryl may be optionally substituted by up to four substituents in any position, whereby each substituent is independently selected from: H, F, Cl, Br, I, R 20 , CF 3 , CF 2 R 7 , CF 2 CF 2 , CC1 3 , CC1 2 R 7 , CC1 2 CC1 2 R 7 , NR 18 R 19 , NR 18 COR 20 , NR ⁇ 8 SO 2 R 20 , OR 20 , OCF 3 , OCF 2 R 20 , OCF 2 CF 2 R 20 , OCOR 20 , OSO 2 R 20 , OPO(OR 20 ) 2 , SR 20 , SCF 3 , SCF 2 R 20 , SCF 2 CF 2 R 20 , SCOR 20 , SO 3 R 20 , S0 2 NR ⁇ 8 R ⁇ 9 , PO(OR 20 ) 3 , PO(
  • R 20 is H, C ⁇ _ 8 straight chain or branched alkyl, C 3 . 8 cycloalkyl, C 4-9 alkylcycloalkyl, C 2 . 8 alkenyl, aryl or alkylaryl; Ris and R 19 are each independently selected from H, C ⁇ -8 straight chain or branched alkyl, C 2 . 8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl, or CH 2 aryl, wherein each moiety within said C ⁇ -8 straight chain or branched alkyl, C 2 .
  • alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl or CH 2 aryl may be optionally substituted by up to four substituents in any position, whereby each substituent is independently selected from: F, Cl, Br, I, CF 3 , CC1 3 , CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , NH 2 , NHCH 3 , N(CH 3 ) 2 , NHC 2 H 5 , N(C 2 H 5 ) 2 , NHC 3 H 7 , N(C 3 H 7 ) 2 , NHC 4 H 9 , N(C 4 H 9 ) 2 , NHCOH, NHCOCH . , NHC0C 2 H 5 ,
  • OCOC 4 H 9 OS0 2 CH 3 , OS0 2 C 2 H 5 , OS0 2 C 3 H 7 , OS0 2 C 4 H 9 , SH, SCH 3 , SC 2 H 5 , SC 3 H 7 , SC 4 H 7 , SGd ,
  • Ris or R ]9 may together form part of a 5, 6 or 7 membered cyclic stracture, with said stracture being saturated or unsaturated, and further with said stracture containing up to four heteroatoms selected from O, N or S, and further wherein each moiety within said cyclic structure being optionally substituted by up to four substituents in any position independently selected from: F, Cl, Br, I, CF 3 , CC1 3 , CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , NH 2 , NHCH 3; N(CH 3 ) 2 , NHC 2 H 5 , N(C 2 H 5 ) 2 , NHC 3 H 7 , N(C 3 H 7 ) 2 , NHC 4 H 9 , N(C 4 H 9 ) 2 , NHCOH, NHCOCH 3 , NHCOC 2 H 5 , NHCOC 3 H 7 , NHCOQH,, NHS0 2 CH 3 , NHS
  • An aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to
  • Also provided by the present invention are methods for modulating the activity of a human 5-HT 2A serotonin receptor by contacting the receptor with a compound of formula:
  • X is O or S;
  • P is H or CH 3 ;
  • R 8 and R 20 are each independently selected from H, C ⁇ -8 straight chain or branched alkyl, C 3-8 cycloalkyl, C 4 . 9 alkylcycloalkyl, or C 2 . 8 alkenyl;
  • R 15 is H, F, Cl, Br, I, R 20 , CF 3 , CF 2 R 20 , CF 2 CF 2 , CC1 3 , CC1 2 R 20 , CC1 2 CC1 2 R 20 , NR 18 R 19 , NR 19 COR 20 , NR 19 SO 2 R 20 , OR 20 , OCF 3 , OCF 2 R 20 , OCF 2 CF 2 R 20 , OCOR 20 , OSO 2 R 20 , OPO(OR 20 ) 2 , SR 20 , SCF 3 , SCF 2 R 20 , SCF 2 CF 2 R 20 , SCOR 20 , SO 3 R 20 , S0 2 NR 18 R ⁇ 9 , PO(OR 20 ) 3 , PO(OR 20 ) 2 R 20 , N0 2 , CN, CNR 20 (NR 18 R ⁇ 9 ), CNR 19 (SR 20 ), COOR 20 , COSR 20 , CONR ⁇ 8 R ⁇ 9 , with the pro
  • Ris and R ⁇ 9 are each independently selected from H, C]. s straight chain or branched alkyl, C 2 . 8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl, or CH 2 aryl, wherein each moiety within said C ⁇ _ 8 straight chain or branched alkyl, C 2-8 alkenyl or cycloalkyl, or alkylcycloalkyl, or aryl or CH 2 aryl may be optionally substituted by up to four substituents in any position, whereby each substituent is independently selected from: F, Cl, Br, I, CF 3 , CC1_, CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , NH 2 , NHCH 3 , N(CH 3 ) 2 ,
  • OCOC 4 H 9 OS0 2 CH 3 , OS0 2 C 2 H 5 , OS0 2 C 3 H 7 , OS0 2 C 4 H 9 , SH, SCH 3 , SC 2 H 5 , SC 3 H 7 , SC 4 H 7 , SC 4 H 9 ,
  • R 13 , R ⁇ 4 , R 15 , Ri 6 and R each independently selected from the following: F, Cl, Br, I, CF 3 , CC1 3 , CH 3 , C 2 H 5 , C 3 H 7 , C 4 H 9 , NH 2 , NHCH 3 , N(CH 3 ) 2 , NHC 2 H 5 , N(C 2 H 5 ) 2 , NHC 3 H 7 , N(C 3 H 7 ) 2 ,
  • NHC 4 H 9 N(C 4 H 9 ) 2 , NHCOH, NHCOCH 3 , NHCOC 2 H 5 , NHC0C 3 H 7 , NHCOC 4 H 9 , NHS0 2 CH 3 ,
  • An aryl moiety can be a 5 or 6 membered aromatic heterocyclic ring (containing up to 4 hetero atoms independently selected from N, O, or S) or a 6 membered aromatic non-heterocyclic ring or a polycycle.
  • the present invention further provides methods for modulating the activity of a human 5-HT 2A serotonin receptor by contacting the receptor with a compound of formula:
  • R 2 is H or lower alkyl (C M );
  • R 3 is Me, or Et, or halogen;
  • X is either Oxygen or Sulfur;
  • Y is NR ⁇ 5 R 16 , or (CH 2 ) m R 17 , or 0(CH 2 ) n R 17 ;
  • R 4 is lower alkyl (C ⁇ _ 6 );
  • R 16 and R are independently C M alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, C0NR 18 R ⁇ 9 , NR ⁇ 8 R ⁇ 9 , NHCOCH 3 , OCF 3 , SMe, COOR 20 , SO 3 R 20 , S0 2 NR 18 R 19 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 .
  • Ris and R 19 are independently a H or -6 alkyl, or C 2 . 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, CONR 2 ⁇ R 22 , NR 21 R 22 , NHCOCH3, OCF 3 , SMe, COOR 23 , S0 3 R 23 , S0 2 NR 2 ⁇ R 22 , COMe, COEt, CO-lower alkyl, SCF 3 , CN, C 2 . 6 alkenyl, H, halogens, Q.
  • C 3 . 6 cycloalkyl, C alkyl, and aryl or R 18 and R i9 may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from O, N or S and said cyclic structure may be "optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, OCF 3 , SMe, COOR 7 , S0 2 NR 24 R 25 , S0 3 R 26 , NHCOCH 3 , COEt, COMe, or halogen;
  • R 20 and R 23 are each independently selected from H or C !-6 alkyl;
  • R 21 and R 22 are each independently are independently a H, or C 1-5 alkyl, or C 2- 6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF 3 , OEt, CC1 3 , Me, N0 2 , OH, OMe, SMe, COMe, CN, COOR 20 , SO 3 R 20 , COEt, NHCOCH 3 , or aryl.
  • An aryl moiety can be a 5 or 6 membered aromatic hetero-cyclic ring (containing up to 4 hetero atoms independently selected from N, O, or S) or a 6 membered aromatic non-heterocyclic ring or a polycycle.
  • the present invention further provides methods for modulating the activity of a human 5-HT 2A serotonin receptor by contacting the receptor with a compound of formula:
  • Ar is a phenyl ring optionally substituted with up to five groups selected from the group consisting of halogen, OR 7 , OH, NRs 9 , carboxy, CN, alkoxycarbonyl, straight chain or branched C M alkyl -C(p) 3j or -0-C(p) 3 where p is halogen;
  • Rs and R 9 are independentiy a H, or C M alkyl, or C 2-6 alkenyl, or cycloalkyl, or aryl, or CH 2 aryl group and each said group may be optionally substituted by up to four substituents in any position independently selected from halogen, CF 3 , OCF 3 , OEt, CC1 3 , Me, N0 2 , OH, OMe, SMe, COMe, CN, COOR 10 , SO 3 R 10 , COEt, NHCOCH 3 , or aryl; or
  • R 8 and R 9 may form part of a 5, 6 or 7 membered cyclic structure which may be either saturated or unsaturated and that may contain up to four heteroatoms selected from 0, N or S and said cyclic structure may be optionally substituted by up to four substituents in any position independently selected from CF 3 , CC1 3 , Me, N0 2 , OH, OMe, OEt, OCF 3 , SMe, COORio, SOzNRsRc,, SO 3 R 10 , NHCOCH 3 , COEt, COMe, or halogen;
  • R 7 is H or C alkyl; Rio is H or CM alkyl;
  • R 2 is H, straight chain or branched C alkyl, C 2 . 6 alkenyl, or cycloalkyl; iv) R 3 is halogen, carboxy, CN, alkoxycarbonyl, straight chain or branched CM alkyl, C 2 _
  • R 4 is C alkyl, C 2 . 6 alkenyl, or cycloalkyl;
  • R n and R 12 are each independently H, straight chain or branched C alkyl, C 2 . 6 alkenyl, or cycloalkyl; wherein:
  • R ⁇ 3 , R ⁇ 4 , Ris, Ri 6 and R are each independently H, halogen, CN, NR 8 R 9 , COORio, SRio, straight chain or branched C ⁇ -6 alkyl, C -6 alkenyl, C 2 . 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl, wherein each of said alkoxycarbonyl, straight chain or branched C M alkyl, C 2 . 6 alkenyl, C 2 .
  • 6 alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , ORio, NRsR . , halogen, -C(p) 3; or -0-C(p) 3 where p is halogen, and said cycloalkyl, aryl, alkylaryl, arylalkyl or heteroaryl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position;
  • Q 2 is straight chain or branched C M alkyl, C 2 . 6 alkenyl, C 2 . ⁇ alkynyl, cycloalkyl, aryl, alkylaryl, arylalkyl, wherein each of said cycloalkyl, aryl, alkylaryl, or arylalkyl groups can be optionally substituted with up to four substituents in any position selected from N0 2 , OR 7 , halogen, -C(p) 3> or -0-C(p) 3 where p is halogen, or an aliphatic or aromatic heterocycle, and said cycloalkyl, aryl, alkylaryl, and arylalkyl groups can alternatively or additionally be optionally substituted with up to four alkyl substituents in any position; or a pharmaceutically acceptable salt.
  • the compounds are selected from compounds and combinations and subcombinations of compounds selected from the group consisting of:
  • the present invention provides the foregoing compounds, and combinations and subcombinations thereof.
  • the present invention provides compounds useful as inverse agonists for 5-HT 2A receptors stracturally represented as follows:
  • compositions comprising each of the compounds ofthe invention.
  • Odier preferred compounds provided by the present invention and useful in the methods disclosed herein include, but are not limited to:
  • R 3 is Br or Cl; t is lower alkyl; R n isH,F,ClorCF 3 ;
  • R 12 is H, F or Cl
  • Specifically preferred compounds of Formula (LX) above include the following:
  • the amine 12-1 can be converted to the corresponding isocyanate 12-4 by die action of phosgene or a suitable phosgene equivalent, e.g. friphosgene, in an inert solvent such as a halocarbon in the presence of an organic base such as triethylamine (i.e., TEA) or diisopropylethylamine (i.e., DIEA).
  • phosgene or a suitable phosgene equivalent e.g. friphosgene
  • Isocyanate 12-4 reacts with amines of the general formula 12-5 in an analogous fashion to that described above in Scheme 12-1 to give urea 12-6.
  • This approach allows for diverse groups to be introduced for the R 2 or R 3 group based on the starting amine 12-5 (Scheme 12-2).
  • Arnines of general formula 12-5 are also readily converted to activated isocyanate equivalents of general fonnula 12-8 by the sequential action of carbonyldiimidazole and metiiyl iodide in tefrahydrofuran and acetonitrile respectively (R.A. Batey et al, Tetrahedron Lett., (1998), 39, 6267-6270.) Reaction of 12-8 with amine 12-1 in an inert solvent such as a halocarbon would yield the requisite ureas of general formula 12-3 (Scheme 12-4).
  • Amine 12-1 may be monomethylated according to the procedure of J. Barluenga et al,
  • Carbamates of general formula 12-12 can be obtained in a similar manner via a variety of synthetic manipulations, all of which would be familiar to one skilled in the art.
  • the reaction of amine 12-1 witii chloroformates (see Org. Syn. Coll. Vol. IN, (1963), 780) of general formula 12-13 in an inert solvent such as ether or halocarbon in the presence of a tertiary base such as friethylamine or ethylisopropylamine readily yields the requisite carbamates of general formula 12-12 (Scheme 12-6).
  • Chloroformates of general formula 12-13 not commercially available may be readily prepared from the corresponding alcohol 12-15 in an inert solvent such as toluene, chlorocarbon or etiier by the action of excess phosgene (see Org. Syn. Coll. Vol. Ill, (1955), 167) (Scheme 12-9).
  • Amide compounds of the general formula 12-16 can be obtained via a variety of synthetic manipulations, all of which would be familiar to one skilled in the art.
  • the reaction of amine 12-1 with acid chlorides (see Org. Syn. Coll. N, (1973), 336) of general formula 12-17 to yield the desired amides 12-16 is readily achieved in an inert solvent such as chloroform or dichloromethane in the presence of an organic base such as friethylamine or emyldiisopropylamine (Scheme 12-10).
  • the corresponding acids of general formula 12-19 may be coupled with dicyclohexylcarbodiimide (DCC)/hydroxybenzofriazole (HOBT) (see W. Konig et al, Chem. Ber., (1970), 103, 788) or hydroxybenzotriazole (HOBT)/2-(lH-benzotriazole-l-yl)-l, 1,3,3- tetramethyluronium hexafluorophosphate (HBTU) (see M.
  • DCC dicyclohexylcarbodiimide
  • HOBT hydroxybenzofriazole
  • HBTU 1,3,3- tetramethyluronium hexafluorophosphate
  • a third aspect of the present invention provides a compound of Formula (I) or a solvate or physiologically functional derivative thereof for use as a tiierapeutic agent, specifically as a modifier ofthe activity of the serotonin 5-HT 2A receptor.
  • Modifiers ofthe activity ofthe serotonin 5- HT 2A receptor are believed to be of potential use for the treatment or prophylaxis of CNS, gastrointestinal, cardiovascular, and inflammatory disorders.
  • Compounds of die Formula (I) may be adieri concludedred by oral, sublingual, parenteral, rectal, or topical administration.
  • physiologically acceptable salts of the compounds may also be formed and used as therapeutic agents.
  • Different amounts of the compounds of Formula (I) will be required to achieve the desired biological effect. The amount will depend on factors such as the specific compound, the use for which it is intended, the means of adminisfration, and the condition of the treated individual.
  • a typical dose may be expected to fall in the range of 0.001 to 200 mg per kilogram of body weight of die treated individual. Unit does may contain from 1 to 200 mg of the compounds of Formula (1) and may be administered one or more times a day, individually or in multiples.
  • the salt or solvate of a compound of Formulas (I) the dose is based on the cation (for salts) or the unsolvated compound.
  • a fourth aspect of the present invention provides pharmaceutical compositions, comprising at least one compound of Formula (I) and/or an acceptable salt or solvate thereof (e.g., a pharmaceutically acceptable salt or solvate) as an active ingredient combined with at least one carrier or excipient (e.g., pharmaceutical carrier or excipient).
  • Pharmaceutical compositions may be used in die treatment of clinical conditions for which a modifier of the activity of the serotonin 5-HT 2A receptor is indicated, particularly where the active ingredient is preferentially selective for the 5-HT 2A receptor over the 5-HT 2A receptor, and most particularly where the active ingredient is also an inverse agonist at the 5-HT 2A receptor.
  • At least one compound of Formula (I) may be combined with the carrier in either solid or liquid form in a unit dose formulation.
  • the pharmaceutical carrier must be compatible with the other ingredients in tiie composition and must be tolerated by the individual recipient.
  • Other physiologically active ingredients may be inco ⁇ orated into the pharmaceutical composition of the invention if desired, and if such ingredients are compatible with the other ingredients in the composition.
  • Formulations may be prepared by any suitable method, typically by uniformly mixing the active compound(s) with liquids or finely divided solid carriers, or both, in the required proportions, and then, if necessary, forming the resulting mixture into a desired shape.
  • liquid preparations for oral administration may be in the form of solutions, emulsions, aqueous or oily suspensions, and syrups.
  • the oral preparations may be in the form of dry powder that can be reconstituted with water or another suitable liquid vehicle before use. Additional additives such as suspending or emulsifying agents, non-aqueous vehicles (including edible oils), preservatives, and flavorings and colorants may be added to the liquid preparations.
  • Parenteral dosage forms may be prepared by dissolving the compound ofthe invention in a suitable liquid vehicle and filter sterilizing the solution before filling and sealing an appropriate vial or ampoule. These are just a few examples ofthe many appropriate methods well known in the art for preparing dosage forms.
  • the fifth aspect of the present invention provides for the use of a compound of formula (I) in the preparation of a medicament for the treatment of a medical condition for which a modifier ofthe activity ofthe serotonin 5-HT 2A receptor is indicated.
  • the sixth aspect of the present invention provides for a method of treatment of a clinical condition of a mammal, such as a human, for which a modifier of the activity of the serotonin 5-HT 2A receptor is indicated, which comprises the administration to the mammal of a therapeutically effective amount of a compound of Formula (I) or a physiologically acceptable salt, solvate, or physiologically functional derivative thereof.
  • Mass spectra were recorded on a Micromass PlatformTM LC with Gilson HPLC.
  • Infra-red spectra were recorded on a Nicolet AvatarTM 360 FT-IR. Melting points were recorded on a Electrothermal IA9200TM apparatus and are uncorrected. Proton nuclear magnetic resonance spectra were recorded on a BrukerTM 300MHz machine. Chemical shifts are given with respect to teframethylsilane. In the text the following abbreviations are used; s (singlet), d (doublet), t (triplet), m (multiplet) or combinations thereof. Chemical shifts are quoted in parts per million (ppm) and with coupling constants in Hertz.
  • HPLC (250 ⁇ L; GF 254 ).
  • HPLC was recorded either on a HP ChemstationTM 1100 HPLC using a Hichrom 3.5 C18 reverse phase column (50mm x 2.1mm i.d.). Linear gradient elution over 5 minutes - 95% water (+0.1% TFA) / 5% acetonitrile (+0.05% TFA) down to 5% water / 95% acetonitrile. Flow rate 0.8mL/min (Method A); or on a Hichrom 3.5 C18 reverse phase column (100mm x 3.2mm i.d.).
  • HPLC retention time 3.28 mins (Method A).
  • Tic Rf 0.4 (EtOAc/hexane).
  • This compound is commercially available from Maybridge Chemical Company, Catalog No. KM04515, under the name N-(3-(4-bromo-l-methylpyrazol-3-yl)phenyl)(((4- trifluoromethoxy)phenyl) amino)carboxamide. :
  • IR: v raax 1684, 1592, 1510, 1253, 1217, 1157, 987, 798, 700 cm "1 .

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EP03705889A 2002-01-23 2003-01-23 Nicht-endogene, konstitutiv aktivierte rezeptoren für menschliches serotonin und modulatoren von kleiner molekülgrösse Pending EP1509505A2 (de)

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PCT/US2003/002059 WO2003062206A2 (en) 2002-01-23 2003-01-23 Small molecule modulators of the 5-ht2a serotonin receptor useful for the prophylaxis and treatment of disorders related thereto

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