WO2012003418A2 - Functionally selective ligands of dopamine d2 receptors - Google Patents

Functionally selective ligands of dopamine d2 receptors Download PDF

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Publication number
WO2012003418A2
WO2012003418A2 PCT/US2011/042734 US2011042734W WO2012003418A2 WO 2012003418 A2 WO2012003418 A2 WO 2012003418A2 US 2011042734 W US2011042734 W US 2011042734W WO 2012003418 A2 WO2012003418 A2 WO 2012003418A2
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Prior art keywords
butoxy
benzo
diazepan
compound
dichlorophenyl
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PCT/US2011/042734
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French (fr)
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WO2012003418A3 (en
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Jian Jin
Bryan Roth
Stephen Frye
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The University Of North Carolina At Chapel Hill
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Priority to US13/807,347 priority Critical patent/US9156822B2/en
Publication of WO2012003418A2 publication Critical patent/WO2012003418A2/en
Publication of WO2012003418A3 publication Critical patent/WO2012003418A3/en

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    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
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    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/472—Non-condensed isoquinolines, e.g. papaverine
    • A61K31/4725—Non-condensed isoquinolines, e.g. papaverine containing further heterocyclic rings
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    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
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    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
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    • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom 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
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    • C07D235/04—Benzimidazoles; Hydrogenated benzimidazoles
    • C07D235/24—Benzimidazoles; Hydrogenated benzimidazoles 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 in position 2
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    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
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    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/08—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms
    • C07D295/084—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
    • C07D295/088—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly bound oxygen or sulfur atoms with the ring nitrogen atoms and the oxygen or sulfur atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
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    • 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
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    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07D471/04—Ortho-condensed systems

Definitions

  • the present invention relates to novel functionally selective ligands of dopamine D 2 receptors, including agonists, antagonists, and inverse agonists.
  • the invention further relates to the use of these compounds in research and for treating central nervous system disorders related to D 2 receptors.
  • a partial agonist possesses a lower degree of intrinsic efficacy and partially activates all cellular responses induced by an agonist Antagonists, according to this schema, are neutral entities which possess no intrinsic activity but are able to block the receptor and preclude activation by full or partial agonists (Furchgott, Advances in Drug Research (Harper N and Simmonds A eds) Academic Press, New York., pp 21- 55 (1966)).
  • GPCR G-protein coupled receptor
  • GPCR G-protein coupled receptor
  • the present invention relates to novel, functionally selective ligands of dopamine D 2 receptors.
  • the compounds of the invention When evaluated in multiple D 2 binding and functional assays, the compounds of the invention showed different activity profiles ranging from full agonists to antagonists to inverse agonists.
  • the compounds of the invention are ⁇ -arrestin biased and, in certain embodiments, also are inverse agonists of Gj-regulated pathways. These pharmacologically active compounds exhibit fewer side effects and are useful in research to study D 2 receptor function and its role in psychosis and for the treatment of CNS disorders.
  • the invention provides a compound of formula I:
  • m is 0 or 1 ;
  • n 0, 1, or 2;
  • q 1 or 2;
  • X is O, NH, or CH 2 ;
  • Y is N or CH
  • Z is C, CH, CH 2 , cycloalkyl, aryl, or heteroaryl
  • Aii and Ar 2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
  • the invention provides a compound of formula II:
  • n 0, 1, or 2;
  • q 1 or 2;
  • X is O, NH, or CH 2 ;
  • Y is N or CH
  • Z is C, CH, CH 2 , cycloalkyl, aryl, or heteroaryl
  • Ari and Ar 2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl; or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
  • the invention provides a compound of formula VI:
  • n 0, 1, or 2;
  • q 1 or 2;
  • X is O, NH, or CH 2 ;
  • Y is N or CH
  • Z is C, CH, CH 2 , cycloalkyl, aryl, or heteroaryl
  • Ari and Ar 2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
  • the invention further relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of the invention and a pharmaceutically acceptable carrier.
  • the invention also relates to a method of modulating the activity of a D 2 dopamine receptor comprising contacting the receptor with a compound of the invention.
  • the invention additionally relates to a method of treating a central nervous system disorder associated with D 2 dopamine receptors in a subject (e.g., a psychiatric, neurological, pituitary, or endocrine disorder), comprising delivering to the subject a therapeutically effective amount of the compound of the invention.
  • a central nervous system disorder associated with D 2 dopamine receptors e.g., a psychiatric, neurological, pituitary, or endocrine disorder
  • the invention further relates to the use of a compound of the invention for treating a central nervous system disorder associated with D 2 dopamine receptors in a subject.
  • the invention further relates to the use of a compound of the invention for stimulating D 2 dopamine receptors in a ⁇ -arrestin biased manner.
  • Figures 1A-1F show that UNC 9975 exhibits potent antipsychotic-like activity in mouse hyperlocomotion studies.
  • Figure 2 shows that UNC9975 and UNC0006 induce catalepsy in p-arrestin-2 knockout mice but not in wild-type littermates.
  • a can mean one or more than one.
  • a cell can mean a single cell or a multiplicity of cells.
  • compositions of this invention means the composition can contain additional components as long as the additional components do not materially alter the composition.
  • materially altered refers to an increase or decrease in the therapeutic effectiveness of the composition of at least about 20% or more as compared to the effectiveness of a composition consisting of the recited components.
  • terapéuticaally effective amount refers to that amount of a composition, compound, or agent of this invention that imparts a modulating effect, which, for example, can be a beneficial effect, to a subject afflicted with a disorder, disease or illness, including improvement in the condition of the subject (e.g., in one or more symptoms), delay or reduction in the progression of the condition, prevention or delay of the onset of the disorder, and/or change in clinical parameters, disease or illness, etc., as would be well known in the art.
  • a therapeutically effective amount or effective amount can refer to the amount of a composition, compound, or agent that improves a condition in a subject by at least 5%, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.
  • Treat” or “treating” or “treatment” refers to any type of action that imparts a modulating effect, which, for example, can be a beneficial effect, to a subject afflicted with a disorder, disease or illness, including improvement in the condition of the subject (e.g., in one or more symptoms), delay or reduction in the progression of the condition, and/or change in clinical parameters, disease or illness, etc., as would be well known in the art.
  • a "central nervous system disorder associated with D 2 dopamine receptors” refers to any disorder in which modulation (either an increase or a decrease) of one or more activities of a D 2 dopamine receptor in a subject results in the treatment of one or more symptoms of the disorder in the subject,
  • “Pharmaceutically acceptable,” as used herein, means a material that is not biologically or otherwise undesirable, i.e., the material can be administered to an individual along with the compositions of this invention, without causing substantial deleterious biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained.
  • the material would naturally be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject, as would be well known to one of skill in the art ⁇ see, e.g., Remington's Pharmaceutical Science; 21 st ed. 2005).
  • Exemplary pharmaceutically acceptable carriers for the compositions of this invention include, but are not limited to, sterile pyrogen-free water and sterile pyrogen-free physiological saline solution.
  • Concurrently means sufficiently close in time to produce a combined effect (that is, concurrently can be simultaneously, or it can be two or more events occurring within a short time period before or after each other).
  • the administration of two or more compounds "concurrently” means that the two compounds are administered closely enough in time that the presence of one alters the biological effects of the other.
  • the two compounds can be administered in the same or different formulations or sequentially. Concurrent administration can be carried out by mixing the compounds prior to administration, or by administering the compounds in two different formulations, for example, at the same point in time but at different anatomic sites or using different routes of administration.
  • ⁇ -arrestin biased refers to a compound that binds the D 2 dopamine receptor, is a partial or full agonist of the ⁇ -arrestin pathway, and is either neutral or an inverse agonist of the Gj-regulated cAMP pathway.
  • neutral refers to a compound that binds to a receptor but does not stimulate a particular receptor-linked pathway.
  • inverse agonist refers to a compound that binds to a receptor and reduces constitutive activity of a particular receptor-linked pathway.
  • alkyl denotes a straight or branched hydrocarbon chain containing 1- 12 carbon atoms, e.g., 1-6 or 1-4 carbon atoms.
  • alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.
  • substituted alkyl an alkyl in which an atom of the alkyl is substituted with, for example, a carbon, nitrogen, sulfur, oxygen, silicon, or halogen atom, or alternatively a nitrogen, sulfur, oxygen, or halogen atom.
  • the term encompasses substituents on alkyl, alkenyl, alkynyl, and cycloalkyl groups.
  • substituents that can be attached to any atom of the alkyl group in a "substituted alkyl” include cyclyl groups, heterocyclyl groups; aryl groups, heteroaryl groups, amino groups, amido groups, nitro groups, cyano groups, azide groups, hydroxy groups, alkoxy groups, acyloxy groups, thioalkoxy groups, acyl thioalkoxy groups, halogen groups, sulfonate groups, sulfonamide groups, ester groups, carboxylic acids, oxygen ⁇ e.g., a carbonyl group), and sulfur ⁇ e.g., a thiocarbonyl group).
  • Substituents also include any chemical functional group that imparts improved water-solubility to the molecule ⁇ e.g., carboxylic acid, carboxylic ester, carboxamido, morpholino, piperazinyl, imidazolyl, thiomorpholino, or tetrazolyl groups; both unsubstituted and substituted).
  • cycloalkyl denotes a monocyclic saturated carbocyclic group containing 3-8 carbon atoms, e.g., 3-6 carbon atoms.
  • Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.
  • alkoxy denotes an oxygen linked to an alkyl or substituted alkyl as defined above.
  • halo and halogen refer to any radical of fluorine, chlorine, bromine or iodine.
  • ring and "ring system” refer to a ring comprising the delineated number of atoms, said atoms being carbon or, where indicated, a heteroatom such as nitrogen, oxygen or sulfur.
  • the ring itself, as well as any substituents thereon, can be attached at any atom that allows a stable compound to be formed.
  • aryl refers to an aromatic 5-8 membered monocyclic or 8-12 membered bicyclic ring system wherein 0, 1, 2, or 3 atoms of each ring can be substituted by a substituent.
  • the term also includes aromatic bicyclic ring systems in which a hydrogen atom has been added to one, two, or three of the ring carbons in one of the rings (e.g., a partially saturated ring). Examples of aryl groups include phenyl, naphthyl and the like.
  • heteroaryl refers to an aromatic 5-8 membered monocyclic or 8-12 membered bicyclic ring system comprising 1-3 heteroatoms if monocyclic or 1-6 heteroatoms if bicyclic, said heteroatoms selected from O, N, or S, wherein 0, 1, 2 or 3 atoms of each ring can be substituted by a substituent.
  • the term also includes aromatic bicyclic ring systems in which a hydrogen atom has been added to one, two, or three of the ring carbons in one of the rings (e.g., a partially saturated ring).
  • heteroaryl groups include pyridyl, furyl or furanyl, benzofuranyl, imidazolyl, benzimidazolyl, pyrimidinyl, thiophenyl or thienyl, benzothiophenyl, quinolinyl, isoquinolinyl, dihydroquinolinyl, dihydroisoquinolinyl, naphthyridinyl, dihydronaphthyridinyl, quinazolinyl, indolyl, indazolyl, thiazolyl, benzothiazolyl, oxazinyl, benzooxazinyl, oxazolyl, benzooxazolyl, dihydrobenzodioxinyl, and the like.
  • Suitable substituents for aryl and heteroaryl groups are the same as the substituents for alkyl groups.
  • the present invention provides novel, functionally selective ligands of dopamine D 2 receptors.
  • the compounds of the invention are ⁇ -arrestin biased.
  • the compounds are at least partial agonists of the ⁇ -arrestin pathway.
  • the compounds are full agonists of the ⁇ -arrestin pathway.
  • the compounds also are neutral or inverse agonists of the Gj-regulated cAMP pathway. These compounds represent the first functionally selective ⁇ -arrestin-biased dopamine D 2 receptor ligands to exhibit antipsychotic activity in vivo.
  • the invention provides a compound of formula I:
  • n 0 or 1
  • q 1 or 2;
  • n 0, 1, or 2;
  • X is O, NH, or CH 2 ;
  • Y is N or CH
  • Z is C, CH, CH 2 , cycloalkyl, aryl, or heteroaryl
  • Ari and Ar 2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
  • Ar 2 is an unsubstituted or substituted bicyclic aryl or heteroaryl.
  • Z is C, CH, CH 2 , cycloalkyl, monocyclic aryl, or monocyclic heteroaryl.
  • the bond indicated as refers to the bond attached to Z and not to any other bond, e.g., not between the two carbon atoms that are present when q is 2.
  • the compound of the invention has the structure of formula II:
  • n 0, 1, or 2;
  • q 1 or 2;
  • X is O, NH, or CH 2 ; Y is N or CH;
  • Z is C, CH, CH 2 , cycloalkyl, aryl, or heteroaryl
  • Arj and Ar 2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
  • the compound of the invention has the structure of formula
  • n 0, 1, or 2;
  • X is O, NH, or CH 2 ;
  • Y is N or CH
  • Ari and Ar 2 are independently an unsubstituted or substituted monocyclic or bicyclic heteroaryl
  • n is 1. In a further embodiment, n is 0. In another embodiment, X is O. In another embodiment, X is NH.
  • Ax ⁇ is selected from the group consisting of phenyl, substituted phenyl (e.g., chlorophenyl, dichlorophenyl, fluorophenyl, trifluoromethylphenyl, methylphenyl, cyanophenyl, methoxyphenyl, ethoxyphenyl, isopropoxyphenyl), 2,3-dihydro-lH-benzo[d]imidazolyl, benzo[d]oxazol-2(3H)-one, 1H- benzo[d]imidazolyl, benzo[d]oxazolyl, IH-indazolyl, IH-indazolyl, IH-indolyl, benzofuranyl, benzo[b]thiophenyl, benzo[d]thiazolyl, naphthalenyl, isoquinolinyl, quinazolinyl, quinolin
  • Ar 2 is selected from the group consisting of 3,4-dihydroquinolin-2(lH)-one, 3,4-dihydro-l ,8-naphthyridin-2(lH)-one, quinolin-2(lH)-one, 3,4-dihydroisoquinolin-l(2H)-one, benzo jthiazole, lH-benzo[ ⁇ /Jimidazol- 2(3H)-one, lH-indazole, and benzo[d]oxazol-2(3H)-one, or any subgroup selected from the list.
  • Ari is dichlorophenyl or methoxyphenyl and Ar 2 is quinolinone, dihydroquinolinone, naphthyridinone, or benzothiazole.
  • ⁇ is dichlorophenyl, methoxyphenyl, ethoxyphenyl, or isopropyloxyphenyl and Ar 2 is quinolinone, dihydroquinolinone, naphthyridinone, benzothiazole, or benzoimidazolone.
  • Ar 2 is not naphthyridinone, dihydronaphthyridinone, quinolinone, dihydroquinolinone, tetrahydropyridoazepinone, or iso indole.
  • the compounds of the invention have the structure of formula
  • the compound of formula II is selected from the group of compounds shown in Tables 1 and 2.
  • the compound of the invention has the structure of formula
  • n 0, 1, or 2;
  • q 1 or 2;
  • X is O, NH, or C3 ⁇ 4;
  • Y is N or CH
  • Z is C, CH, CH 2 , cycloalkyl, aryl, or heteroaryl
  • Ari and Ar 2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
  • Ar 2 is an unsubstituted or substituted bicyclic aryl or heteroaryl.
  • Z is C, CH, CH 2 , cycloalkyl, monocyclic aryl, or monocyclic heteroaryl.
  • n is 0. In a different embodiment, n is 1. In another embodiment, X is O or NH.
  • Ari is selected from the group consisting of substituted phenyl ⁇ e.g., dichlorophenyl, methoxyphenyl), 2H- benzo[b][l,4]oxazin-3(4H)-one, 2,3-dihydrobenzo[b][l,4]dioxinyl, and benzo[d]oxazol-2(3H)- one, or any subgroup selected from the list.
  • Ax ⁇ is dichlorophenyl.
  • Ar 2 is selected from the group consisting of IH-indazole, benzo[d]thiazole, imidazol-2(3H)-one, 3,4-dihydroisoquinolin- l(2H)-one, 3,4-dihydroquinolin-2(lH)-one, lH-benzo[d]imidazol-2(3H)-one, 3,4-dihydro-l,8- naphthyridin-2(lH)-one, quinolin-2(lH)-one, 2-methylphenylacetamide, and benzo[d]oxazol- 2(3H)-one, or any subgroup selected from the list.
  • Ar 2 is selected from the group consisting of IH-indazole, benzo[d]thiazole, and imidazol-2(3H)-one.
  • Ari is dichlorophenyl or dihydrobenzodioxinyl and Ar 2 is dihydroquinolinone, benzothiazole, or indazole.
  • Ari is dichlorophenyl, methoxyphenyl, or dihydrobenzodioxinyl and Ar 2 is dihydroquinolinone, benzothiazole, indazole, benzoimidazolone, or quinolinone.
  • Y is N or CH
  • n is 0 or 1
  • X is O
  • Ari is dichlorophenyl
  • Ar 2 is selected from the group consisting of IH-indazole, benzo[d]thiazole, and lH-benzoimidazol-2(3H)-one.
  • Ar 2 is not naphthyridinone, dihydronaphthyridinone, quinolinone, dihydroquinolinone, tetrahydropyridoazepinone, or isoindole.
  • the compound of formula VI is selected from the group of compounds shown in Tables 3 and 4.
  • the compounds of this invention include all pharmaceutically acceptable salt forms thereof.
  • such salts include those derived from pharmaceutically acceptable inorganic and organic acids and bases.
  • suitable acid salts include, without limitation, acetate, adipate, alginate, aspartate, benzoate, butyrate, citrate, fumarate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2- hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, nicotinate, nitrate, oxalate, palmoate, pectinate, persulfate, hydroxynapthoate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate.
  • Other acids such as oxalic, while not in themselves pharmaceutically acceptable, can be
  • Salts derived from appropriate bases include, without limitation, alkali metal (e.g., sodium, potassium), alkaline earth metal (e.g., magnesium and calcium), ammonium and N- (alkyl)4 + salts.
  • the compounds of this invention include deuterium-substituted versions of the disclosed compounds, e.g., compounds comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more deuterium atoms.
  • the compounds of the invention include prodrugs of the compounds that are converted to the active compound in vivo.
  • the compound can be modified to enhance cellular permeability (e.g., by esterification of polar groups) and then converted by cellular enzymes to produce the active agent.
  • Methods of masking charged or reactive moieties as a pro-drug are known by those skilled in the art ⁇ see, e.g., P. Korgsgaard-Larsen and H. Bundgaard, A Textbook of Drug Design and Development, Reading U.K., Harwood Academic Publishers, 1991).
  • prodrug refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formula, for example, by hydrolysis in blood, see, e.g., T. Higuchi and V. Stella, Prodrugs as Novel delivery Systems, Vol. 14 of the A.C.S. Symposium Series and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated by reference herein. See also U.S. Patent No. 6,680,299.
  • Exemplary prodrugs include a prodrug that is metabolized in vivo by a subject to an active drug having an activity of the compounds as described herein, wherein the prodrug is an ester of an alcohol or carboxylic acid group, if such a group is present in the compound; an amide of an amine group or carboxylic acid group, if such groups are present in the compound; a urethane of an amine group, if such a group is present in the compound; an acetal or ketal of an alcohol group, if such a group is present in the compound; a N-Mannich base or an imine of an amine group, if such a group is present in the compound; or a Schiff base, oxime, acetal, enol ester, oxazolidine, or thiazolidine of a carbonyl group, if such a group is present in the compound, such as described, for example, in U.S. Patent No. 6,680,324 and U.S
  • prodrug refers to those prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and/or other animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable risk/benefit ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
  • the compounds in Tables 1 and 3 were designed and synthesized as novel, functionally selective ligands of dopamine D 2 receptors to elucidate the key signal transduction pathways essential for antipsychotic efficacy and the undesired ones associated with side-effects. These novel compounds were evaluated in multiple D 2 binding and functional assays: D 2 radioligand binding, D 2 cAMP GloSensor, D 2 ERK phosphorylation (p-ERK), and/or D 2 ⁇ - arrestin 2 translocation (Tango). In the D 2 cAMP GloSensor assay, these compounds showed different activity profiles ranging from full agonists to antagonists to inverse agonists of the Gj- regulated cAMP pathway.
  • these compounds showed significantly attenuated antipsychotic-like activity and increased catalepsy in p-arrestin-2 knockout mice.
  • These novel, pharmacologically active compounds are useful agents for studying dopamine receptor function and signaling pathways, as well as for treating CNS disorders.
  • the compounds may have advantageous pharmacokinetic characteristics for treating CNS disorders, including a longer half life in brain and higher brain :plasma ratio over 24 h compared to other antipsychotic compounds.
  • the receptor binding pattern of the compounds e.g., relative low potency at histamine receptors suggests that the compounds may exhibit a lower risk for side effects, such as sedation and/or cognitive impairment.
  • one aspect of the invention relates to a method of modulating the activity of a D 2 dopamine receptor comprising contacting the receptor with a compound of the invention.
  • the D 2 dopamine receptor may be present in a cell, an isolated cell membrane or an artificial membrane.
  • the cell may be a cultured cell, a cell isolated from a subject, or a cell in a subject (e.g., an animal model of disease or a patient).
  • the receptor may be a wild-type or modified receptor and may be naturally-occurring or recombinantly expressed.
  • modulating is intended to encompass any type of change in one or more activities of the receptor, including increasing or decreasing the activity of one or more receptor-linked signaling pathways.
  • Many of the compounds of the invention display potent antipsychotic activity in animal models in vivo and are thus predicted to possess antipsychotic actions in humans and other animals.
  • the compounds would thus be effective for treating diseases including, but not limited to, a variety of psychiatric disorders including: schizophrenia, schizoaffective disorder, schizophreniform disorder, bipolar disorder, mania, manic psychosis, Tourette's syndrome and Tourette-like disorders, obsessive-compulsive disorder, depression with psychotic features, psychosis not otherwise specified (Psychosis NOS) and additional affective and non-affective psychiatric disorders.
  • the compounds would be effective against a number of neurological disorders including Huntington's Disease, Parkinsonian Psychosis, Alzheimer's Disease and associated psychosis and various organic brain syndromes. Further, the compounds would be effective at a variety of autism spectrum and autistic disorders. Given the D -receptor functional selectivity, the compounds would also be effective against pituitary diseases including pituitary adenomas and prolactinomas as well as endocrinological disorders including, but not limited to, galactorrhea.
  • the compounds have significant and unexpected advantages over conventional antipsychotic drugs in that they are predicted to effectively treat psychotic disorders without appreciable extra-pyramidal side effects (EPS), having minimal propensity for elevating serum prolactin and with a potentially lower risk of inducing tardive dyskinesia.
  • EPS extra-pyramidal side effects
  • the compounds Given the low affinities for Hi -histamine and partial agonism for 5-HT 2C serotonin receptors the compounds are expected to differ from approved atypical antipsychotic drugs by virtue of a lower propensity to induce weight gain and associated metabolic adverse consequences (diabetes, hyperlipidemia, hyperglycemia, hypercholesterolemia, hypertension and so on (Kroeze et al, Neuropsychopharmacology 28:519 (2003)).
  • the compounds also lack agonist activity at 5-HT 2 B serotonin receptors and hERG potassium channels and are thus predicted to have a low incidence of cardiovascular side-effects including prolongation of the Q-T interval and drug-induced valvular heart disease (Roth, N. Engl. J. Med. 356:6 (2007)). Given the affinities of certain compounds for 5-HT 6 and 5-HT 7 receptors the compounds are expected to have potential cognition-enhancing and antidepressant actions (Abbas et al, Psychopharmacology (Berl) 205: 119 (2009); Woolley et al, Neuropharmacology 41 :210 (2001)). Thus, these compounds are likely to also be effective in treating bipolar depression, unipolar depression, and a variety of anxiety disorders as well as disorders such as Alzheimer's Disease, dementia, Niemann-Pick Disorder, and other late-life dementias.
  • one aspect of the invention relates to a method of treating a central nervous system disorder associated with D 2 dopamine receptors in a subject, comprising delivering to the subject a therapeutically effective amount of the compound of the invention.
  • the disorder associated with D 2 dopamine receptors is a psychiatric, neurological, pituitary, or endocrine disorder.
  • one or more of the compounds of the invention is administered to the subject as needed to treat a disorder.
  • the compound can be administered continuously or intermittently.
  • the compound is administered to the subject more than once a day or once every 1, 2, 3, 4, 5, 6, or 7 days.
  • the compound is administered to the subject no more than once a week, e.g., no more than once every two weeks, once a month, once every two months, once every three months, once every four months, once every five months, once every six months, or longer.
  • the compound is administered using two or more different schedules, e.g., more frequently initially (for example to build up to a certain level, e.g., once a day or more) and then less frequently (e.g., once a week or less).
  • the compound can be administered by any discontinuous administration regimen.
  • the compound can be administered not more than once every three days, every four days, every five days, every six days, every seven days, every eight days, every nine days, or every ten days, or longer.
  • the administration can continue for one, two, three, or four weeks or one, two, or three months, or longer.
  • the compound after a period of rest, can be administered under the same or a different schedule.
  • the period of rest can be one, two, three, or four weeks, or longer, according to the pharmacodynamic effects of the compound on the subject.
  • the compound of the invention can be delivered to the subject by any suitable route, e.g., oral, rectal, buccal (e.g., sub-lingual), vaginal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), topical (i.e., both skin and mucosal surfaces, including airway surfaces) and transdermal administration.
  • the compound is delivered to the subject at a dose that is effective to treat the disorder.
  • the effective dosage will depend on many factors including the gender, age, weight, and general physical condition of the subject, the severity of the disorder, the particular compound or composition being administered, the duration of the treatment, the nature of any concurrent treatment, the carrier used, and like factors within the knowledge and expertise of those skilled in the art.
  • the compound is administered at a dose of about 0.001 to about 10 mg/kg body weight, e.g., about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg/kg.
  • the dose can be even lower, e.g., as low as 0.0005 or 0.0001 mg/kg or lower.
  • the dose can be even higher, e.g., as high as 20, 50, 100, 500, or 1000 mg/kg or higher.
  • the present invention encompasses every sub-range within the cited ranges and amounts.
  • the compound of the invention is delivered to a subject concurrently with an additional therapeutic agent.
  • the additional therapeutic agent can be delivered in the same composition as the compound or in a separate composition.
  • the additional therapeutic agent can be delivered to the subject on a different schedule or by a different route as compared to the compound.
  • the additional therapeutic agent can be any agent that provides a benefit to the subject.
  • Further agents include, without limitation, anti-psychotics, antidepressants, agents for neurological disorders, and chemotherapeutic agents.
  • anti-psychotic agents include, without limitation, for psychosis: prochloroperazine, chlorpromazine, droperidol, fluphenazine, periciazine, perphenazine, thiothixene, triflupromazine, haloperidol, molindone, pimozide, and thioridazine: and for schizophrenia: olanzapine, paliperidone, aripiprazole, quetiapine, risperidone, clozapine, flupenthixene, iloperidone, loxapine, mesoridazine, promazine, reserpine, thioridazine, zuclopenthixol, asenapine, levomepromazine, ziprasidone, molindone, pimozide, and thioridazine.
  • agents for depression and related disorders include, without limitation, for depression: modafinil, desvenlafaxine, nortriptyline, seleginline, amoxapine, citalopram, clomipramine, fluoxetine, isocarboxazid, mapriotiline, nefazodone, niacin, nortriptyline, phenelzine, protriptyline, tranylcypromine, trazodone, trimipramine, desipramine, imipramine, methylphenidate, buproprion, alprazolam, amitriptyline, chlordiazepoxide, perphenazine, doxepin, mirtazapine, 5-hydroxytryptophan, duloxetine, escitalopram, venlafaxine, desvenlafaxine, paroxetine, fluoxetine, olanzapine, 1-methylfolate, amitriptyline, sertraline, fluvoxamine
  • agents for neurological disorders include, without limitation, for Alzheimer's disease: caprylidene, donepezil, galantamine, tacrine, vitamin E, ergoloid mesylates, rivastigmine; for Parkinson's disease: nadolol, zonisamide, amantadine, apomorphine, belladonna, benztropine, biperiden, bromocriptine, carbidopa, entacapone, levodopa, pergolide mesylate, pramipexole, procyclidine, rasagiline, ropinirole, rotiotine, scopolamine, tolcapone, trihexylphenidyl, rivastigmine, seleginline; for Huntington's disease: baclofen, pregabalin, tetrabenazine, methylprednisolone, desvenlafaxine, nortriptyline; and for dementia: haloperidol and
  • chemotherapeutic agents include, without limitation, acivicin, aclarubicin, acodazole hydrochloride, acronine, adozelesin, aldesleukin, altretamine, ambomycin, ametantrone acetate, aminoglutethimide, amsacrine, anastrozole, anthramycin, asparaginase, asperlin, azacytidine, azetepa, azotomycin, batimastat, benzodepa, bicalutamide, bisantrene hydrochloride, bisnafide dimesylate, bizelesin, bleomycin sulfate, brequinar sodium, bropirimine, busulfan, cactinomycin, calusterone, caracemide, carbetimer, carboplatin, carmustine, carubicin hydrochloride, carzelesin, cedefingol, chloramb
  • chemotherapeutic agents include, but are not limited to, 20-epi- 1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein- 1; prostatic carcinoma antiandrogen; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-C
  • Suitable subjects are generally mammalian subjects.
  • mammalian subjects includes, but is not limited to, humans, non-human primates, cattle, sheep, goats, pigs, horses, cats, dog, rabbits, rodents (e.g., rats or mice), etc.
  • Human subjects include neonates, infants, juveniles, adults and geriatric subjects.
  • the subject is a human subject that has a CNS disorder associated with D 2 dopamine receptors.
  • the subject used in the methods of the invention is an animal model of a CNS disorder associated with D 2 dopamine receptors.
  • the subject can be a subject "in need of the methods of the present invention, e.g., in need of the therapeutic effects of the inventive methods.
  • the subject can be a subject that is experiencing a CNS disorder associated with D 2 dopamine receptors and/or is anticipated to experience a CNS disorder associated with D 2 dopamine receptors, and the methods and compositions of the invention are used for therapeutic and/or prophylactic treatment.
  • the compounds of the invention described above can be formulated for administration in a pharmaceutical carrier in accordance with known techniques. See, e.g., Remington, The Science And Practice of Pharmacy (21 st ed. 2005).
  • the compound is typically admixed with, inter alia, an acceptable carrier.
  • the carrier must, of course, be acceptable in the sense of being compatible with any other ingredients in the formulation and must not be deleterious to the patient.
  • the carrier can be a solid or a liquid, or both, and can be formulated with the compound as a unit-dose formulation, for example, a tablet, which can contain from 0.01% or 0.5% to 95% or 99% by weight of the compound.
  • One or more compounds can be incorporated in the formulations of the invention, which can be prepared by any of the well known techniques of pharmacy comprising admixing the components, optionally including one or more accessory ingredients.
  • compositions of the invention include those suitable for oral, rectal, topical, buccal (e.g., sub-lingual), vaginal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), topical (i.e., both skin and mucosal surfaces, including airway surfaces) and transdermal administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated and on the nature of the particular active compound which is being used.
  • Formulations suitable for oral administration can be presented in discrete units, such as capsules, cachets, lozenges, or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as a solution or a suspension in an aqueous or nonaqueous liquid; or as an oil-in-water or water-in-oil emulsion.
  • Such formulations can be prepared by any suitable method of pharmacy which includes the step of bringing into association the compound and a suitable carrier (which can contain one or more accessory ingredients as noted above).
  • the formulations of the invention are prepared by uniformly and intimately admixing the compound with a liquid or finely divided solid carrier, or both, and then, if necessary, shaping the resulting mixture.
  • a tablet can be prepared by compressing or molding a powder or granules containing the compound, optionally with one or more accessory ingredients.
  • Compressed tablets can be prepared by compressing, in a suitable machine, the compound in a free-flowing form, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, and/or surface active/dispersing agent(s).
  • Molded tablets can be made by molding, in a suitable machine, the powdered compound moistened with an inert liquid binder.
  • Formulations suitable for buccal (sub-lingual) administration include lozenges comprising the compound in a flavored base, usually sucrose and acacia or tragacanth; and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacia.
  • Formulations of the present invention suitable for parenteral administration comprise sterile aqueous and non-aqueous injection solutions of the compound, which preparations are preferably isotonic with the blood of the intended recipient. These preparations can contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient.
  • Aqueous and non-aqueous sterile suspensions can include suspending agents and thickening agents.
  • the formulations can be presented in unit/dose (e.g., in a syringe or other injection device) or multi-dose containers, for example sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or water-for-injection immediately prior to use.
  • sterile liquid carrier for example, saline or water-for-injection immediately prior to use.
  • Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules and tablets of the kind previously described.
  • an injectable, stable, sterile composition comprising one or more compounds, in a unit dosage form in a sealed container.
  • the compound is provided in the form of a lyophilizate which is capable of being reconstituted with a suitable pharmaceutically acceptable carrier to form a liquid composition suitable for injection thereof into a subject.
  • the unit dosage form typically comprises from about 0.001 mg to about 10 grams of the compound.
  • a sufficient amount of emulsifying agent which is physiologically acceptable can be employed in sufficient quantity to emulsify the compound in an aqueous carrier.
  • emulsifying agent is phosphatidyl choline.
  • Formulations suitable for rectal administration are preferably presented as unit dose suppositories. These can be prepared by admixing the compound with one or more conventional solid carriers, for example, cocoa butter, and then shaping the resulting mixture.
  • one or more conventional solid carriers for example, cocoa butter
  • Formulations suitable for topical application to the skin preferably take the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil.
  • Carriers which can be used include petroleum jelly, lanoline, polyethylene glycols, alcohols, transdermal enhancers, and combinations of two or more thereof.
  • Formulations suitable for transdermal administration can be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. Formulations suitable for transdermal administration can also be delivered by iontophoresis (see, for example, Pharm. Res. 3:318 (1986)) and typically take the form of an optionally buffered aqueous solution of the compound. Suitable formulations comprise citrate or bis ⁇ tris buffer (pH 6) or ethanol/water and contain from 0.1 to 0.2 M active ingredient.
  • compositions can be prepared from the compounds disclosed herein, such as aqueous base emulsions.
  • the composition will contain a sufficient amount of pharmaceutically acceptable emulsifying agent to emulsify the desired amount of the compound.
  • Particularly useful emulsifying agents include phosphatidyl cholines and lecithin.
  • the pharmaceutical compositions can contain other additives, such as pH-adjusting additives.
  • useful pH-adjusting agents include acids, such as hydrochloric acid, bases or buffers, such as sodium lactate, sodium acetate, sodium phosphate, sodium citrate, sodium borate, or sodium gluconate.
  • the compositions can contain microbial preservatives.
  • Useful microbial preservatives include methylparaben, propylparaben, and benzyl alcohol. The microbial preservative is typically employed when the formulation is placed in a vial designed for multidose use.
  • additives that are well known in the art include, e.g., detackifiers, anti-foaming agents, antioxidants (e.g., ascorbyl palmitate, butyl hydroxy anisole (BHA), butyl hydroxy toluene (BHT) and tocopherols, e.g., a-tocopherol (vitamin E)), preservatives, chelating agents (e.g., EDTA and/or EGTA), viscomodulators, tonicifiers (e.g., a sugar such as sucrose, lactose, and/or mannitol), flavorants, colorants, odorants, opacifiers, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof.
  • detackifiers e.g., anti-foaming agents
  • antioxidants e.g., ascorbyl palmitate, butyl hydroxy anisole (BHA), butyl hydroxy to
  • the additive can also comprise a thickening agent.
  • suitable thickening agents can be those known and employed in the art, including, e.g., pharmaceutically acceptable polymeric materials and inorganic thickening agents.
  • Exemplary thickening agents for use in the present pharmaceutical compositions include polyacrylate and polyacrylate co-polymer resins, for example poly-acrylic acid and poly-acrylic acid/methacrylic acid resins; celluloses and cellulose derivatives including: alkyl celluloses, e.g., methyl-, ethyl- and propyl-celluloses; hydroxyalkyl- celluloses, e.g., hydroxypropyl-celluloses and hydroxypropylalkyl-celluloses such as hydroxypropyl-methyl-celluloses; acylated celluloses, e.g., cellulose-acetates, cellulose- acetatephthallates, cellulose-acetatesuccinates and hydroxypropylmethyl-cellulose phthallates; and salts thereof
  • thickening agents as described above can be included, e.g., to provide a sustained release effect.
  • the use of thickening agents as aforesaid will generally not be required and is generally less preferred.
  • Use of thickening agents is, on the other hand, indicated, e.g., where topical application is foreseen.
  • the present invention provides liposomal formulations of the compounds disclosed herein.
  • the technology for forming liposomal suspensions is well known in the art.
  • the compound When the compound is in the form of an aqueous-soluble material, using conventional liposome technology, the same can be incorporated into lipid vesicles. In such an instance, due to the water solubility of the compound, the compound will be substantially entrained within the hydrophilic center or core of the liposomes.
  • the lipid layer employed can be of any conventional composition and can either contain cholesterol or can be cholesterol-free.
  • the compound of interest is water-insoluble, again employing conventional liposome formation technology, the compound can be substantially entrained within the hydrophobic lipid bilayer which forms the structure of the liposome.
  • the liposomes which are produced can be reduced in size, as through the use of standard sonication and homogenization techniques.
  • the liposomal formulations containing the compound disclosed herein can be lyophilized to produce a lyophilizate which can be reconstituted with a pharmaceutically acceptable carrier, such as water, to regenerate a liposomal suspension.
  • Mass spectra (MS) and HPLC (UV 254 nM or ELSD) data for all compounds were recorded on an 1100 LC/MS system (Agilent Technology Corporation) using a 4.6x50 mm column (CenturySIL C-18 AQ + , 5 ⁇ ) with a linear gradient 30-90% (v/v) acetonitrile-water with 0.035% trifluoroacetic acid over 5 min with a flow rate of 3.5 mL/min.
  • Analytical TLC was performed using 2.5 x 5 cm plates coated with 0.25 mm of silica gel GF254. Column chromatography was performed on silica gel G (200-300 mesh) with reagent grade solvents.
  • a well-dried flask was first charged with l-bromo-2,3-dichlorobenzene (10 g, 44 mmol) and piperazine (4.55 g, 53 mmol), which was evacuated and backfilled with N 2 through a balloon under gentle warming (40°C). Toluene was charged and the mixture was bubbled with N 2 for 10 min, then BINAP (822 mg, 1.32 mmol) and Pd 2 dba 3 (403 mg, 0.44 mmol) were added to the mixture.
  • Triethylamine in acetone was added dropwise to the solution of 2,3- dihydrobenzo[b][l,4]dioxine-5-carboxylic acid (360 mg, 2 mmol) in water and acetone at 0°C.
  • ethyl carbonochloridate was added dropwise.
  • the reaction mixture was stirred for 1 h then it was poured into ice water and extracted with Et 2 0. The combined organic extracts were washed with brine, dried with MgS0 4 , and concentrated in vacuo.
  • Cells stably expressing D 2 receptors were plated in 15-cm dishes (in DMEM containing 10% fetal bovine serum and grown to 90% confluence. Then, cells were washed with PBS, pH 7.4, and harvested by scraping into PBS, pH 7.4. Harvested cells were centrifuged at 1,000 x g for 10 min, then hypotonically lysed by resuspension in ice-cold 50 mM HEPES, 1% BSA, pH 7.4. Membranes were isolated by centrifugation at 21,000 x g for 20 min. The supernatant was removed and the membrane pellets were assayed for D2 ligand-stimulated GTP-gamma-S binding or stored at -80°C until used for radioligand binding assays.
  • Membranes prepared as above were resuspended to 1 ⁇ g protein/ ⁇ (measured by Bradford assay using BSA as standard), and 50 ⁇ were added to each well of a polypropylene 96-well plate containing (per well): 50 ⁇ of buffer (20 mM HEPES, 10 mM MgCl 2 , 1 mM EDTA, 1 mM EGTA, 100 mM N-methyl-D-gluconate, pH 7.4), 50 ⁇ of 1.5 nM [ 3 H]N- methylspiperone (final concentration 0.3 nM) and reference or D2 test ligand at various concentrations ranging from 50 pM to 50 ⁇ (final concentrations ranging from 10 pM to 10 ⁇ , triplicate determinations for each concentration of D 2 test ligand).
  • buffer 20 mM HEPES, 10 mM MgCl 2 , 1 mM EDTA, 1 mM EGTA, 100 mM N-methyl-D-gluconate
  • Residual [ 3 H]N-methylspiperone binding to filtered membranes was plotted as a function of log [reference] or log [D 2 test ligand] and the data were regressed using the one-site competition model built into Prism 4.0 (GraphPad software).
  • HEK293T cells co-expressing the cAMP biosensor GloSensor-22F (Promega) and hD 2 receptors were seeded (10,000 cells/20 ⁇ /well) into white, clear-bottom, tissue culture plates in HBSS, 10% FBS, 20 mM HEPES, pH 7.4. After a one- to two-hour recoveiy, cells were treated with 10 ⁇ of 3X test or reference drug prepared in HBSS, 10% FBS, 20 mM HEPES, pH 7.4. After 30 minutes, cells were challenged with 10 ⁇ of 1,200 nM (4X) isoproterenol in 8% (4X) GloSensor reagent.
  • Luminescence per well per second was read on a Wallac TriLux microbeta plate counter. Data were normalized to the isoproterenol response (100%) and the maximal quinpirole-induced inhibition thereof (0%) and regressed using the sigmoidal dose- response function built into GraphPad Prism 4.0. Notably, HEK293T cells expressing the GloSensor-22F alone (no hD 2 ) were assayed in parallel and displayed no inhibition of isoproterenol-stimulated cAMP, either by quinpirole or by the test compounds, suggesting that the effect observed in hD 2 -expressing cells was due to compound acting via the recombinant receptor.
  • Immunofluorescence Automatic multichannel pipetters were used for liquid handling and multichannel vacuum manifolds for aspirations. Each tested concentration was typically measured in triplicate or quadruplicate. For concentration curves, half-log-dilutions were used. Drug dilutions were prepared in stimulation medium (serum-free medium, 100 mg/L ascorbic acid). 100 ⁇ dopamine and 1 ⁇ PMA (phorbol 12-myristate 13 -acetate) were used as positive controls. Medium only was used as a negative control. Specificity of the response was confirmed by pretreatment for 5 min with 10 ⁇ of the antagonist spiperone and in experiments using wild type CHO cells.
  • stimulation medium serum-free medium, 100 mg/L ascorbic acid
  • PMA phorbol 12-myristate 13 -acetate
  • Plates were incubated in 100 ⁇ , ⁇ blocking buffer (PBS/Ca, 5% goat serum, 0.1% Triton X-100) for 1 h at room temperature and then in blocking buffer containing rabbit phospho-Thr202/p-Tyr204-ERK antibody (Cell Signaling 9101) at 1 :1000 at 4°C overnight. Cells were washed 3X for 5-10 min with 250 ⁇ wash buffer (PBS/Ca, 0.03% Triton X-100).
  • PBS/Ca 5% goat serum, 0.1% Triton X-100
  • Plates were incubated with 50 ⁇ Alexa-594 coupled goat anti-rabbit IgG (Invitrogen) at 1 :250, 5 ⁇ g/mL Hoechst 33342 nuclear stain, and 25 ⁇ g/mL concanavaline-Alexa488 conjugate (ConA, Invitrogen) in blocking buffer for 2 h at room temperature. Plates were washed three times with 250 ⁇ ⁇ ⁇ washing buffer, post-fixed for 10 min in fixing buffer, washed with 250 ⁇ PBS/Ca, filled with 200 nL/well PBS/Ca, sealed with transparent adhesive plate seals, and stored at 4°C.
  • Alexa-594 coupled goat anti-rabbit IgG Invitrogen
  • ConA concanavaline-Alexa488 conjugate
  • Microscopy and Image Analysis Plates were scanned with a high-content automated microscopic system (BD Pathway 855) using a 20x objective and a 2 x 2 image montage setting.
  • the Alexa594 light path was used for the target signal, Alexa488 for whole cell staining, and Hoechst for the nuclear staining.
  • Well-averaged individual cell-based measurements were exported to Excel and cell- free background intensity was subtracted from the whole cell intensity. Concentration curves were analyzed in GraphPad Prism by fitting against a sigmoidal dose-response model and normalization to the dopamine curve.
  • the cAMP (Glo-Sensor) assay shows that the majority of the tested compounds are not agonists for the Gj-coupled signaling pathway and that 14 of the 43 compounds function as inverse agonists for the Gj-coupled signaling pathway (i.e., have a negative E max ).
  • the p-ERK assay confirms that the majority of compounds lack agonism via canonical G-protein-dependent pathways.
  • 2 and 3 had moderate to high binding affinity ( j's: 0.6 - 250 nM) at 5HT 2A , 5HT 2B , 5HT 2C , and 5HTi A , but were significantly less potent in functional assays (Ca 2+ mobilization fluorometric imaging plate reader (FLIPR) or cAMP biosensor).
  • 2 and 3 were antagonists at 5HT 2A , 5HT 2B , and 5HT 2C and weak agonists (EC 50 's > 1 ⁇ ) at 5HTi A .
  • 2 and 3 had moderate affinities to Hi -histamine receptor (Xj's ⁇ 10 nM) but were less potent in Hi functional assays.
  • 2 and 3 display a similar G protein-coupled receptor selectivity profile as aripiprazole (Table 7).
  • Neutral ligands are either antagonists or are inactive; NA: not applicable.
  • Kj, IC 50 , pA 2 , or EC 50 values are the average of at least two duplicate experiments with standard deviation (SD) values that are 3-fold less than the average.
  • mice All experiments were approved by the Institutional Animal Care and Use Committees at the University of North Carolina, Chapel Hill and Duke University. C57BL/6J and P-arrestin-2 knockout (Bohn et al, Science 286:2495 (1999)) mice were housed under standard conditions - 12 hour light/dark cycle and food and water provided ad libitum. Adult, age-matched male and female C57BL/6J and P-arrestin-2 knockout drug naive mice were used for all behavioral testing.
  • Locomotor activity was assessed in photocell-based activity chambers under standardized environmental conditions, using an AccuScan activity monitor (AccuScan Instruments, Columbus, OH) with a 25.8 x 25.8 cm Plexiglas chamber and a beam spacing of 1.52 cm as described (Abbas et al, J. Neurosci. 29:7124 (2009)). Mice were injected (i.p.) with vehicle (0.9% saline/0.2% acetic acid) , aripiprazole (0.10, 0.25, 0.50 or 2.0 mg/kg), or UNC9975 (0.25, 0.5 and 2.0 mg/kg) and placed into the open field.
  • vehicle i.p.
  • aripiprazole 0.10, 0.25, 0.50 or 2.0 mg/kg
  • UNC9975 0.25, 0.5 and 2.0 mg/kg
  • mice were initially injected (i.p.) with vehicle (0.9% saline/0.2% acetic acid), aripiprazole, U C0006 (5 mg/kg), UNC9975 (5 mg/kg) or haloperidol (2 mg/kg) and returned to their home cage.
  • vehicle 0.9% saline/0.2% acetic acid
  • U C0006 5 mg/kg
  • UNC9975 5 mg/kg
  • haloperidol haloperidol
  • UNC9975 exhibited potent antipsychotic-like activity in mouse hyperlocomotion studies which is attenuated in p-arrestin-2 knockout mice .
  • locomotion responses in inbred C57BL/6 mice are shown as 5 minute binned intervals to different doses (i.p.) of UNC9975 or vehicle followed 30 min later by 3 mg/kg D-Amphetamine (AMPH, i.p.).
  • AMPH i.p.
  • Fig. IB total distance traveled after D-Amphetamine administration (30-70 min time interval) is shown.
  • C57BL/6 mice were given vehicle or different doses of UNC9975 or aripiprazole 30 min prior to AMPH treatment.
  • n 8 animals/group. *, p ⁇ 0.05 versus vehicle + 3 mg/kg D- Amphetamine group.
  • locomotor activities are shown as 5 min binned intervals of wild-type (WT) or P-arrestin-2 knockout ( ⁇ - ⁇ 2 KO) littermate mice given vehicle or different doses of UNC9975 followed 30 min later with 6 mg/kg phencyclidine (PCP, i.p.).
  • UNC9975 and U C0006 induced catalepsy in p-arrestin-2 knockout mice but not in wild-type littermates.
  • wild-type and p-Arrestin-2 knockout littermate mice were administered (i.p.) vehicle, 5.0 mg/kg UNC9975, 5.0 mg/kg UNC0006, 5.0 mg/kg aripiprazole, or 2.0 mg/kg haloperidol.
  • novel D 2 ⁇ -arrestin-biased agonists were designed, synthesized and characterized, and demonstrated to have unique atypical antipsychotic drug-like activities in vivo. These novel compounds represent the first functionally selective ⁇ -arrestin- biased dopamine D 2 receptor ligands to exhibit antipsychotic activity in vivo. These findings show that based on the arrestin-bias of these compounds, ⁇ -arrestin may be an important contributor to both antipsychotic drug efficacy and antipsychotic side effects.
  • the disclosed compounds were discovered as unique, ⁇ -arrestin-biased functionally selective D 2 ligands.
  • both UNC0006 and UNC9975 are potent partial agonists which induce D 2 receptors-mediated ⁇ - arrestin recruitment and signaling, and simultaneously are potent inverse agonists at Gi-dependent signaling.
  • the in vivo atypical antipsychotic drug-like activity of UNC9975 indicated a strict requirement for p-arrestin-2 for both full antipsychotic activity and protection against motoric side-effects.
  • this ⁇ -arrestin-biased ligand shows a potent ability to suppress both D-amphetamine and phencyclidine-induced hyperlocomotion in mice, indicating that the compound possesses antipsychotic drug-like activities in vivo.
  • the antipsychotic drug-like activity of UNC9975 was attenuated in ⁇ -3 ⁇ 8 ⁇ -2 knockout mice, indicating p-arrestin-2 is required in vivo for full activity.
  • aripiprazole With the exception of aripiprazole, all FDA approved typical and atypical antipsychotic medications (e.g., haloperidol, chlorpromazine, clozapine, risperidone) share the common property of antagonizing D 2 -mediated G protein-dependent and independent signaling (Masri et al, Proc. Natl. Acad. Sci. USA 105:13656 (2008)). Indeed, typical and atypical antipsychotic drugs, with the exception of aripiprazole, are inverse agonists at Gj-mediated signaling and antagonists at arrestin-ergic pathways.
  • typical and atypical antipsychotic drugs are inverse agonists at Gj-mediated signaling and antagonists at arrestin-ergic pathways.

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Abstract

The present invention relates to novel functionally selective ligands of dopamine D2 receptors, including agonists, antagonists, and inverse agonists. The invention further relates to the use of these compounds for treating central nervous system disorders related to D2 receptors.

Description

Functionally Selective Ligands of Dopamine D2 Receptors
Statement of Priority
[0001] This application claims the benefit of U.S. Provisional Application Serial No. 61/360,951, filed July 2, 2010, the entire contents of which is incorporated by reference herein.
Statement of Government Support
[0002] Aspects of this invention were made with government support under National Institute of Mental Health Grant Nos. U19MH082441S1 and U19MH082441. The United States Government has certain rights to this invention.
Field of the Invention
[0003] The present invention relates to novel functionally selective ligands of dopamine D2 receptors, including agonists, antagonists, and inverse agonists. The invention further relates to the use of these compounds in research and for treating central nervous system disorders related to D2 receptors.
Background of the Invention
[0004] Classical notions of receptor pharmacology imply that when a ligand interacts with a receptor, only a unitary outcome is possible. According to these models of receptor pharmacology, a full or partial agonist can activate only a single signal transduction pathway while an antagonist can only block the actions of an agonist. The key theoretical construct underlying this model was the concept of intrinsic efficacy (Furchgott, Advances in Drug Research (Harper N and Simmonds A eds) Academic Press, New York, pp 21-55 (1966)). According to this conceptualization, a full agonist has maximum intrinsic efficacy and maximally stimulates all cellular responses induced by ligand binding. A partial agonist possesses a lower degree of intrinsic efficacy and partially activates all cellular responses induced by an agonist Antagonists, according to this schema, are neutral entities which possess no intrinsic activity but are able to block the receptor and preclude activation by full or partial agonists (Furchgott, Advances in Drug Research (Harper N and Simmonds A eds) Academic Press, New York., pp 21- 55 (1966)). An extension of this model has been that a single G-protein coupled receptor (GPCR) interacts with a single G protein subtype (e.g., Gi, Gs, Gq, Go and so on) and that full and partial agonists activate only a single transduction pathway.
[0005] For many decades, however, it has been clear that these simplistic notions of GPCR (also known as 7-transmembrane receptor or 7-TM) function cannot account for the myriad of actions induced by agonist or antagonist binding. This was first convincingly demonstrated for the serotonin (5-hydroxytryptamine; 5-HT) and dopamine (DA) families of receptors where it has been demonstrated that: (1) agonists differentially activate distinct signal transduction pathways (Beaulieu et al, J. Neurosci. 27:881 (2007); Berg et al, Mol. Pharmacol. 54:94 (1998); Ghosh et al, J. Med. Chem. 39:549 (1996); Kilts et al, J. Pharmacol. Exp. Ther. 307: 1179 (2002); Mottola et al, J. Pharmacol. Exp. Ther. 307: 1166 (2002); Parrish et al, J. Neurochem. 95:1575 (2005); Roth and Chuang, Life Sci. ¥7:1051 (1987); Urban et al, J. Pharmacol. Exp. Ther. 320:1 (2007)); (2) antagonists can possess negative intrinsic activity (e.g., function as inverse agonists) or be silent and thereby block the actions of an inverse agonist (e.g., neutral antagonists) (Barker et al, J. Biol. Chem. 269:11687 (1994); Burstein et al, J. Pharmacol. Exp. Ther. 375:1278 (2005); Chidiac et al, Mol Pharmacol. 45:490 (1994); Gilliland and Alper, Naunyn Schmiedebergs Arch. Pharmacol. 3(57:498 (2000); Rauser et al, J. Pharmacol. Exp. Ther. 299:83 (2001)); (3) antagonists can also induce receptor down regulation (Peroutka and Snyder, Science 270:88 (1980)) and receptor internalization in vitro (Berry et al, Mol. Pharmacol. 50:306 (1996)) and in vivo (Gray and Roth, Brain Res. Bull 56:441 (2001); Willins et al, Ann. NY Acad. Sci. 861:121 (1998); Willins et al, euroscience 91:599 (1999)) - properties typically associated with agonists.
[0006] The processes by which agonists and antagonists differentially modulate signaling pathways and receptor trafficking have been variously dubbed 'functional selectivity', 'biased agonism' or 'agonist-directed trafficking'. Currently, the preferred term for the process by which ligands can differentially activate signaling pathways mediated via a single GPCR is functional selectivity (Urban et al, J. Pharmacol. Exp. Ther. 320:1 (2007)). Functional selectivity has been demonstrated for more than a dozen GPCRs (reviewed in (Urban et al, Neuropsychopharmacology 32:67 (2007)) and is, likely, a ubiquitous phenomenon. In particular, functional selectivity has been widely observed among DA receptor subtypes.
[0007] It is desirable to identify novel, functionally selective ligands of dopamine D2 receptors to elucidate the key signal transduction pathways essential for antipsychotic efficacy and the undesired ones associated with side-effects. Such compounds would also be useful agents for treating central nervous system (CNS) disorders including schizophrenia and depression.
Summary of the Invention
[0008] The present invention relates to novel, functionally selective ligands of dopamine D2 receptors. When evaluated in multiple D2 binding and functional assays, the compounds of the invention showed different activity profiles ranging from full agonists to antagonists to inverse agonists. In particular, the compounds of the invention are β-arrestin biased and, in certain embodiments, also are inverse agonists of Gj-regulated pathways. These pharmacologically active compounds exhibit fewer side effects and are useful in research to study D2 receptor function and its role in psychosis and for the treatment of CNS disorders.
[0009] Accordingly, as one aspect, the invention provides a compound of formula I:
Figure imgf000005_0001
wherein m is 0 or 1 ;
n is 0, 1, or 2;
q is 1 or 2;
X is O, NH, or CH2;
Y is N or CH;
Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is a single, double, or triple bond as valencies permit; and
Aii and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0010] In another aspect, the invention provides a compound of formula II:
Figure imgf000005_0002
wherein n is 0, 1, or 2;
q is 1 or 2;
X is O, NH, or CH2;
Y is N or CH;
Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is a single, double, or triple bond as valencies permit; and
Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl; or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0011] In a further aspect, the invention provides a compound of formula VI:
Figure imgf000006_0001
wherein n is 0, 1, or 2;
q is 1 or 2;
X is O, NH, or CH2;
Y is N or CH;
Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is a single, double, or triple bond as valencies permit; and
Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0012] The invention further relates to a pharmaceutical composition comprising the compound of the invention and a pharmaceutically acceptable carrier.
[0013] The invention also relates to a method of modulating the activity of a D2 dopamine receptor comprising contacting the receptor with a compound of the invention.
[0014] The invention additionally relates to a method of treating a central nervous system disorder associated with D2 dopamine receptors in a subject (e.g., a psychiatric, neurological, pituitary, or endocrine disorder), comprising delivering to the subject a therapeutically effective amount of the compound of the invention.
[0015] The invention further relates to the use of a compound of the invention for treating a central nervous system disorder associated with D2 dopamine receptors in a subject.
[0016] The invention further relates to the use of a compound of the invention for stimulating D2 dopamine receptors in a β-arrestin biased manner.
[0017] The present invention is explained in greater detail in the drawings herein and the specification set forth below. Brief Description of the Drawings
[0018] Figures 1A-1F show that UNC 9975 exhibits potent antipsychotic-like activity in mouse hyperlocomotion studies.
[0019] Figure 2 shows that UNC9975 and UNC0006 induce catalepsy in p-arrestin-2 knockout mice but not in wild-type littermates.
Detailed Description of the Invention
[0020] The present invention can be embodied in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. For example, features illustrated with respect to one embodiment can be incorporated into other embodiments, and features illustrated with respect to a particular embodiment can be deleted from that embodiment. In addition, numerous variations and additions to the embodiments suggested herein will be apparent to those skilled in the art in light of the instant disclosure, which do not depart from the instant invention.
[0021] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
[0022] Unless the context indicates otherwise, it is specifically intended that the various features of the invention described herein can be used in any combination.
[0023] Moreover, the present invention also contemplates that in some embodiments of the invention, any feature or combination of features set forth herein can be excluded or omitted.
[0024] To illustrate, if the specification states that a complex comprises components A, B and C, it is specifically intended that any of A, B or C, or a combination thereof, can be omitted and disclaimed singularly or in any combination.
[0025] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference herein in their entirety.
Definitions.
[0026] As used herein, "a," "an," or "the" can mean one or more than one. For example, "a" cell can mean a single cell or a multiplicity of cells.
[0027] Also as used herein, "and/or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or"). [0028] Furthermore, the term "about," as used herein when referring to a measurable value such as an amount of a compound or agent of this invention, dose, time, temperature, and the like, is meant to encompass variations of ± 20%, ± 10%, ± 5%, ± 1%, ± 0.5%), or even + 0.1% of the specified amount.
[0029] The term "consists essentially of (and grammatical variants), as applied to the compositions of this invention, means the composition can contain additional components as long as the additional components do not materially alter the composition. The term "materially altered," as applied to a composition, refers to an increase or decrease in the therapeutic effectiveness of the composition of at least about 20% or more as compared to the effectiveness of a composition consisting of the recited components.
[0030] The term "therapeutically effective amount" or "effective amount," as used herein, refers to that amount of a composition, compound, or agent of this invention that imparts a modulating effect, which, for example, can be a beneficial effect, to a subject afflicted with a disorder, disease or illness, including improvement in the condition of the subject (e.g., in one or more symptoms), delay or reduction in the progression of the condition, prevention or delay of the onset of the disorder, and/or change in clinical parameters, disease or illness, etc., as would be well known in the art. For example, a therapeutically effective amount or effective amount can refer to the amount of a composition, compound, or agent that improves a condition in a subject by at least 5%, e.g., at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%.
[0031] "Treat" or "treating" or "treatment" refers to any type of action that imparts a modulating effect, which, for example, can be a beneficial effect, to a subject afflicted with a disorder, disease or illness, including improvement in the condition of the subject (e.g., in one or more symptoms), delay or reduction in the progression of the condition, and/or change in clinical parameters, disease or illness, etc., as would be well known in the art.
[0032] A "central nervous system disorder associated with D2 dopamine receptors" refers to any disorder in which modulation (either an increase or a decrease) of one or more activities of a D2 dopamine receptor in a subject results in the treatment of one or more symptoms of the disorder in the subject,
[0033] "Pharmaceutically acceptable," as used herein, means a material that is not biologically or otherwise undesirable, i.e., the material can be administered to an individual along with the compositions of this invention, without causing substantial deleterious biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained. The material would naturally be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject, as would be well known to one of skill in the art {see, e.g., Remington's Pharmaceutical Science; 21st ed. 2005). Exemplary pharmaceutically acceptable carriers for the compositions of this invention include, but are not limited to, sterile pyrogen-free water and sterile pyrogen-free physiological saline solution.
[0034] "Concurrently" means sufficiently close in time to produce a combined effect (that is, concurrently can be simultaneously, or it can be two or more events occurring within a short time period before or after each other). In some embodiments, the administration of two or more compounds "concurrently" means that the two compounds are administered closely enough in time that the presence of one alters the biological effects of the other. The two compounds can be administered in the same or different formulations or sequentially. Concurrent administration can be carried out by mixing the compounds prior to administration, or by administering the compounds in two different formulations, for example, at the same point in time but at different anatomic sites or using different routes of administration.
[0035] The term "β-arrestin biased" refers to a compound that binds the D2 dopamine receptor, is a partial or full agonist of the β-arrestin pathway, and is either neutral or an inverse agonist of the Gj-regulated cAMP pathway.
[0036] The term "neutral" refers to a compound that binds to a receptor but does not stimulate a particular receptor-linked pathway.
[0037] The term "inverse agonist" refers to a compound that binds to a receptor and reduces constitutive activity of a particular receptor-linked pathway.
[0038] The term "alkyl" denotes a straight or branched hydrocarbon chain containing 1- 12 carbon atoms, e.g., 1-6 or 1-4 carbon atoms. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, and the like.
[0039] By "substituted alkyl" is meant an alkyl in which an atom of the alkyl is substituted with, for example, a carbon, nitrogen, sulfur, oxygen, silicon, or halogen atom, or alternatively a nitrogen, sulfur, oxygen, or halogen atom. The term encompasses substituents on alkyl, alkenyl, alkynyl, and cycloalkyl groups.
[0040] Examples of substituents that can be attached to any atom of the alkyl group in a "substituted alkyl" include cyclyl groups, heterocyclyl groups; aryl groups, heteroaryl groups, amino groups, amido groups, nitro groups, cyano groups, azide groups, hydroxy groups, alkoxy groups, acyloxy groups, thioalkoxy groups, acyl thioalkoxy groups, halogen groups, sulfonate groups, sulfonamide groups, ester groups, carboxylic acids, oxygen {e.g., a carbonyl group), and sulfur {e.g., a thiocarbonyl group). Substituents also include any chemical functional group that imparts improved water-solubility to the molecule {e.g., carboxylic acid, carboxylic ester, carboxamido, morpholino, piperazinyl, imidazolyl, thiomorpholino, or tetrazolyl groups; both unsubstituted and substituted).
[0041] The term "cycloalkyl" denotes a monocyclic saturated carbocyclic group containing 3-8 carbon atoms, e.g., 3-6 carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.
[0042] The term "alkoxy" denotes an oxygen linked to an alkyl or substituted alkyl as defined above.
[0043] The terms "halo" and "halogen" refer to any radical of fluorine, chlorine, bromine or iodine.
[0044] The terms "ring" and "ring system" refer to a ring comprising the delineated number of atoms, said atoms being carbon or, where indicated, a heteroatom such as nitrogen, oxygen or sulfur. The ring itself, as well as any substituents thereon, can be attached at any atom that allows a stable compound to be formed.
[0045] The term "aryl" refers to an aromatic 5-8 membered monocyclic or 8-12 membered bicyclic ring system wherein 0, 1, 2, or 3 atoms of each ring can be substituted by a substituent. The term also includes aromatic bicyclic ring systems in which a hydrogen atom has been added to one, two, or three of the ring carbons in one of the rings (e.g., a partially saturated ring). Examples of aryl groups include phenyl, naphthyl and the like.
[0046] The term "heteroaryl" refers to an aromatic 5-8 membered monocyclic or 8-12 membered bicyclic ring system comprising 1-3 heteroatoms if monocyclic or 1-6 heteroatoms if bicyclic, said heteroatoms selected from O, N, or S, wherein 0, 1, 2 or 3 atoms of each ring can be substituted by a substituent. The term also includes aromatic bicyclic ring systems in which a hydrogen atom has been added to one, two, or three of the ring carbons in one of the rings (e.g., a partially saturated ring). Examples of heteroaryl groups include pyridyl, furyl or furanyl, benzofuranyl, imidazolyl, benzimidazolyl, pyrimidinyl, thiophenyl or thienyl, benzothiophenyl, quinolinyl, isoquinolinyl, dihydroquinolinyl, dihydroisoquinolinyl, naphthyridinyl, dihydronaphthyridinyl, quinazolinyl, indolyl, indazolyl, thiazolyl, benzothiazolyl, oxazinyl, benzooxazinyl, oxazolyl, benzooxazolyl, dihydrobenzodioxinyl, and the like.
[0047] Suitable substituents for aryl and heteroaryl groups are the same as the substituents for alkyl groups.
Compounds.
[0048] The present invention provides novel, functionally selective ligands of dopamine D2 receptors. In particular, the compounds of the invention are β-arrestin biased. In certain embodiments, the compounds are at least partial agonists of the β-arrestin pathway. In other embodiments, the compounds are full agonists of the β-arrestin pathway. In further embodiments, the compounds also are neutral or inverse agonists of the Gj-regulated cAMP pathway. These compounds represent the first functionally selective β-arrestin-biased dopamine D2 receptor ligands to exhibit antipsychotic activity in vivo.
[0049] In one aspect the invention provides a compound of formula I:
Figure imgf000011_0001
wherein m is 0 or 1;
q is 1 or 2;
n is 0, 1, or 2;
X is O, NH, or CH2;
Y is N or CH;
Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is optionally a single, double, or triple bond as valencies permit; and Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0050] In one embodiment, Ar2 is an unsubstituted or substituted bicyclic aryl or heteroaryl. In another embodiment, Z is C, CH, CH2, cycloalkyl, monocyclic aryl, or monocyclic heteroaryl. The bond indicated as refers to the bond attached to Z and not to any other bond, e.g., not between the two carbon atoms that are present when q is 2.
[0051] In one aspect the compound of the invention has the structure of formula II:
Figure imgf000011_0002
wherein n is 0, 1, or 2;
q is 1 or 2;
X is O, NH, or CH2; Y is N or CH;
Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is optionally a single, double, or triple bond as valencies permit; and Arj and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0052] In one embodiment, the compound of the invention has the structure of formula
III:
Figure imgf000012_0001
wherein n is 0, 1, or 2;
X is O, NH, or CH2;
Y is N or CH; and
Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0053] In another e structure of formula IV:
Figure imgf000012_0002
IV or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0054] In one embodiment for the compound of formula II-IV, n is 1. In a further embodiment, n is 0. In another embodiment, X is O. In another embodiment, X is NH.
[0055] In one embodiment for the compound of formula II-IV, Ax\ is selected from the group consisting of phenyl, substituted phenyl (e.g., chlorophenyl, dichlorophenyl, fluorophenyl, trifluoromethylphenyl, methylphenyl, cyanophenyl, methoxyphenyl, ethoxyphenyl, isopropoxyphenyl), 2,3-dihydro-lH-benzo[d]imidazolyl, benzo[d]oxazol-2(3H)-one, 1H- benzo[d]imidazolyl, benzo[d]oxazolyl, IH-indazolyl, IH-indazolyl, IH-indolyl, benzofuranyl, benzo[b]thiophenyl, benzo[d]thiazolyl, naphthalenyl, isoquinolinyl, quinazolinyl, quinolinyl, and pyridinyl, or any subgroup selected from the list. In a further embodiment, Ari is dichlorophenyl.
[0056] In one embodiment for the compound of formula II-IV, Ar2 is selected from the group consisting of 3,4-dihydroquinolin-2(lH)-one, 3,4-dihydro-l ,8-naphthyridin-2(lH)-one, quinolin-2(lH)-one, 3,4-dihydroisoquinolin-l(2H)-one, benzo jthiazole, lH-benzo[</Jimidazol- 2(3H)-one, lH-indazole, and benzo[d]oxazol-2(3H)-one, or any subgroup selected from the list.
[0057] In one embodiment for the compound of formula II-IV, Ari is dichlorophenyl or methoxyphenyl and Ar2 is quinolinone, dihydroquinolinone, naphthyridinone, or benzothiazole.
[0058] In one embodiment for the compound of formula II-IV, Α is dichlorophenyl, methoxyphenyl, ethoxyphenyl, or isopropyloxyphenyl and Ar2 is quinolinone, dihydroquinolinone, naphthyridinone, benzothiazole, or benzoimidazolone.
[0059] In certain embodiments for the compound of formula II-IV, Ar2 is not naphthyridinone, dihydronaphthyridinone, quinolinone, dihydroquinolinone, tetrahydropyridoazepinone, or iso indole.
[0060] In a further embodi
Figure imgf000013_0001
[0061] In certain embodiments, the compounds of the invention have the structure of formula
Figure imgf000013_0002
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0062] In one embodiment, the compound of formula II is selected from the group of compounds shown in Tables 1 and 2.
Table 1
Figure imgf000013_0003
Figure imgf000014_0001
Figure imgf000015_0001
Table 2
Figure imgf000015_0002
Figure imgf000016_0001
Figure imgf000017_0001
Figure imgf000018_0001
Figure imgf000019_0001
[0063] In one embodiment, the compound of the invention has the structure of formula
Figure imgf000019_0002
wherein n is 0, 1, or 2;
q is 1 or 2;
X is O, NH, or C¾;
Y is N or CH;
Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is optionally a single, double, or triple bond as valencies permit; and
Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
[0064] In one embodiment, Ar2 is an unsubstituted or substituted bicyclic aryl or heteroaryl. In another embodiment, Z is C, CH, CH2, cycloalkyl, monocyclic aryl, or monocyclic heteroaryl.
[0065] In one embodiment for the compound of formula VI, n is 0. In a different embodiment, n is 1. In another embodiment, X is O or NH.
[0066] In one embodiment for the compound of formula VI, Ari is selected from the group consisting of substituted phenyl {e.g., dichlorophenyl, methoxyphenyl), 2H- benzo[b][l,4]oxazin-3(4H)-one, 2,3-dihydrobenzo[b][l,4]dioxinyl, and benzo[d]oxazol-2(3H)- one, or any subgroup selected from the list. In a further embodiment, Ax\ is dichlorophenyl. [0067] In another embodiment for the compound of formula VI, Ar2 is selected from the group consisting of IH-indazole, benzo[d]thiazole, imidazol-2(3H)-one, 3,4-dihydroisoquinolin- l(2H)-one, 3,4-dihydroquinolin-2(lH)-one, lH-benzo[d]imidazol-2(3H)-one, 3,4-dihydro-l,8- naphthyridin-2(lH)-one, quinolin-2(lH)-one, 2-methylphenylacetamide, and benzo[d]oxazol- 2(3H)-one, or any subgroup selected from the list. In a further embodiment, Ar2 is selected from the group consisting of IH-indazole, benzo[d]thiazole, and imidazol-2(3H)-one.
[0068] In another embodiment for the compound of formula VI, Ari is dichlorophenyl or dihydrobenzodioxinyl and Ar2 is dihydroquinolinone, benzothiazole, or indazole.
[0069] In another embodiment for the compound of formula V, Ari is dichlorophenyl, methoxyphenyl, or dihydrobenzodioxinyl and Ar2 is dihydroquinolinone, benzothiazole, indazole, benzoimidazolone, or quinolinone.
[0070] In another embodiment for the compound of formula VI, Y is N or CH, n is 0 or 1, X is O, Ari is dichlorophenyl, and Ar2 is selected from the group consisting of IH-indazole, benzo[d]thiazole, and lH-benzoimidazol-2(3H)-one.
[0071] In certain embodiments for the compound of formula VI, Ar2 is not naphthyridinone, dihydronaphthyridinone, quinolinone, dihydroquinolinone, tetrahydropyridoazepinone, or isoindole.
[0072] In one embodiment, the compound of formula VI is selected from the group of compounds shown in Tables 3 and 4.
Table 3
Figure imgf000020_0001
Figure imgf000021_0001
Figure imgf000022_0001
Figure imgf000023_0001
Table 4
Figure imgf000023_0002
Figure imgf000024_0001
Figure imgf000025_0001
Figure imgf000026_0001
[0073] The compounds of the invention can be synthesized in a similar way as Examples 1 - 51 below and/or according to Feenstra et al, Bioorg. Med. Chem. Lett. 11:2345 (2001) (incorporated herein by reference in its entirety).
[0074] The compounds of this invention include all pharmaceutically acceptable salt forms thereof. Examples of such salts include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acid salts include, without limitation, acetate, adipate, alginate, aspartate, benzoate, butyrate, citrate, fumarate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2- hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, nicotinate, nitrate, oxalate, palmoate, pectinate, persulfate, hydroxynapthoate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate and undecanoate. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, can be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts.
[0075] Salts derived from appropriate bases include, without limitation, alkali metal (e.g., sodium, potassium), alkaline earth metal (e.g., magnesium and calcium), ammonium and N- (alkyl)4+ salts.
[0076] Compounds of the formulae herein include those having quaternization of any basic nitrogen-containing group therein.
[0077] The compounds of this invention include deuterium-substituted versions of the disclosed compounds, e.g., compounds comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more deuterium atoms.
[0078] The discussion herein is, for simplicity, provided without reference to stereoisomerism. Those skilled in the art will appreciate that the compounds of the invention can contain one or more asymmetric centers and thus occur as racemates and racemic mixtures, single optical isomers, individual diastereomers, and diastereomeric mixtures. All such isomeric forms of these compounds are expressly included in the present invention.
[0079] Similarly, compounds of the invention containing a double bond can exist in the form of geometric isomers, which can be readily separated and recovered by conventional procedures. Such isomeric forms are included in the scope of this invention.
[0080] Further, the compounds of the invention include prodrugs of the compounds that are converted to the active compound in vivo. For example, the compound can be modified to enhance cellular permeability (e.g., by esterification of polar groups) and then converted by cellular enzymes to produce the active agent. Methods of masking charged or reactive moieties as a pro-drug are known by those skilled in the art {see, e.g., P. Korgsgaard-Larsen and H. Bundgaard, A Textbook of Drug Design and Development, Reading U.K., Harwood Academic Publishers, 1991).
[0081] The term "prodrug" refers to compounds that are rapidly transformed in vivo to yield the parent compound of the above formula, for example, by hydrolysis in blood, see, e.g., T. Higuchi and V. Stella, Prodrugs as Novel delivery Systems, Vol. 14 of the A.C.S. Symposium Series and in Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated by reference herein. See also U.S. Patent No. 6,680,299. Exemplary prodrugs include a prodrug that is metabolized in vivo by a subject to an active drug having an activity of the compounds as described herein, wherein the prodrug is an ester of an alcohol or carboxylic acid group, if such a group is present in the compound; an amide of an amine group or carboxylic acid group, if such groups are present in the compound; a urethane of an amine group, if such a group is present in the compound; an acetal or ketal of an alcohol group, if such a group is present in the compound; a N-Mannich base or an imine of an amine group, if such a group is present in the compound; or a Schiff base, oxime, acetal, enol ester, oxazolidine, or thiazolidine of a carbonyl group, if such a group is present in the compound, such as described, for example, in U.S. Patent No. 6,680,324 and U.S. Patent No. 6,680,322.
[0082] The term "pharmaceutically acceptable prodrug" (and like terms) as used herein refers to those prodrugs of the compounds of the present invention which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and/or other animals without undue toxicity, irritation, allergic response and the like, commensurate with a reasonable risk/benefit ratio, and effective for their intended use, as well as the zwitterionic forms, where possible, of the compounds of the invention.
Methods.
[0083] The compounds in Tables 1 and 3 were designed and synthesized as novel, functionally selective ligands of dopamine D2 receptors to elucidate the key signal transduction pathways essential for antipsychotic efficacy and the undesired ones associated with side-effects. These novel compounds were evaluated in multiple D2 binding and functional assays: D2 radioligand binding, D2 cAMP GloSensor, D2 ERK phosphorylation (p-ERK), and/or D2 β- arrestin 2 translocation (Tango). In the D2 cAMP GloSensor assay, these compounds showed different activity profiles ranging from full agonists to antagonists to inverse agonists of the Gj- regulated cAMP pathway. In the D2 β-arrestin recruitment (Tango) assay, these compounds showed activity profiles ranging from little agonist activity to full agonist activity of the β- arrestin pathway. In the D2 ERK phosphorylation (p-ERK) assay, these compounds again showed different activity profiles ranging from antagonists to partial agonists that are more efficacious compared to aripiprazole with similar potency. Several β-arrestin biased, functionally selective D2 ligands were efficacious in D-amphetamine-induced and phencyclidine (PCP)- induced hyperlocomotion mouse models and did not induce catalepsy in wild-type mice at pharmacologically active doses. Interestingly, these compounds showed significantly attenuated antipsychotic-like activity and increased catalepsy in p-arrestin-2 knockout mice. These novel, pharmacologically active compounds are useful agents for studying dopamine receptor function and signaling pathways, as well as for treating CNS disorders. Furthermore, the compounds may have advantageous pharmacokinetic characteristics for treating CNS disorders, including a longer half life in brain and higher brain :plasma ratio over 24 h compared to other antipsychotic compounds. Additionally, the receptor binding pattern of the compounds (e.g., relative low potency at histamine receptors) suggests that the compounds may exhibit a lower risk for side effects, such as sedation and/or cognitive impairment.
[0084] Thus, one aspect of the invention relates to a method of modulating the activity of a D2 dopamine receptor comprising contacting the receptor with a compound of the invention. The D2 dopamine receptor may be present in a cell, an isolated cell membrane or an artificial membrane. The cell may be a cultured cell, a cell isolated from a subject, or a cell in a subject (e.g., an animal model of disease or a patient). The receptor may be a wild-type or modified receptor and may be naturally-occurring or recombinantly expressed. The term "modulating" is intended to encompass any type of change in one or more activities of the receptor, including increasing or decreasing the activity of one or more receptor-linked signaling pathways.
[0085] Many of the compounds of the invention display potent antipsychotic activity in animal models in vivo and are thus predicted to possess antipsychotic actions in humans and other animals. The compounds would thus be effective for treating diseases including, but not limited to, a variety of psychiatric disorders including: schizophrenia, schizoaffective disorder, schizophreniform disorder, bipolar disorder, mania, manic psychosis, Tourette's syndrome and Tourette-like disorders, obsessive-compulsive disorder, depression with psychotic features, psychosis not otherwise specified (Psychosis NOS) and additional affective and non-affective psychiatric disorders. Additionally, the compounds would be effective against a number of neurological disorders including Huntington's Disease, Parkinsonian Psychosis, Alzheimer's Disease and associated psychosis and various organic brain syndromes. Further, the compounds would be effective at a variety of autism spectrum and autistic disorders. Given the D -receptor functional selectivity, the compounds would also be effective against pituitary diseases including pituitary adenomas and prolactinomas as well as endocrinological disorders including, but not limited to, galactorrhea.
[0086] The compounds have significant and unexpected advantages over conventional antipsychotic drugs in that they are predicted to effectively treat psychotic disorders without appreciable extra-pyramidal side effects (EPS), having minimal propensity for elevating serum prolactin and with a potentially lower risk of inducing tardive dyskinesia. Given the low affinities for Hi -histamine and partial agonism for 5-HT2C serotonin receptors the compounds are expected to differ from approved atypical antipsychotic drugs by virtue of a lower propensity to induce weight gain and associated metabolic adverse consequences (diabetes, hyperlipidemia, hyperglycemia, hypercholesterolemia, hypertension and so on (Kroeze et al, Neuropsychopharmacology 28:519 (2003)). The compounds also lack agonist activity at 5-HT2B serotonin receptors and hERG potassium channels and are thus predicted to have a low incidence of cardiovascular side-effects including prolongation of the Q-T interval and drug-induced valvular heart disease (Roth, N. Engl. J. Med. 356:6 (2007)). Given the affinities of certain compounds for 5-HT6 and 5-HT7 receptors the compounds are expected to have potential cognition-enhancing and antidepressant actions (Abbas et al, Psychopharmacology (Berl) 205: 119 (2009); Woolley et al, Neuropharmacology 41 :210 (2001)). Thus, these compounds are likely to also be effective in treating bipolar depression, unipolar depression, and a variety of anxiety disorders as well as disorders such as Alzheimer's Disease, dementia, Niemann-Pick Disorder, and other late-life dementias.
[0087] Thus, one aspect of the invention relates to a method of treating a central nervous system disorder associated with D2 dopamine receptors in a subject, comprising delivering to the subject a therapeutically effective amount of the compound of the invention. In certain embodiments, the disorder associated with D2 dopamine receptors is a psychiatric, neurological, pituitary, or endocrine disorder.
[0088] In one embodiment of the invention, one or more of the compounds of the invention is administered to the subject as needed to treat a disorder. The compound can be administered continuously or intermittently. In one embodiment, the compound is administered to the subject more than once a day or once every 1, 2, 3, 4, 5, 6, or 7 days. In another embodiment, the compound is administered to the subject no more than once a week, e.g., no more than once every two weeks, once a month, once every two months, once every three months, once every four months, once every five months, once every six months, or longer. In a further embodiment, the compound is administered using two or more different schedules, e.g., more frequently initially (for example to build up to a certain level, e.g., once a day or more) and then less frequently (e.g., once a week or less). In other embodiments, the compound can be administered by any discontinuous administration regimen. In one example, the compound can be administered not more than once every three days, every four days, every five days, every six days, every seven days, every eight days, every nine days, or every ten days, or longer. The administration can continue for one, two, three, or four weeks or one, two, or three months, or longer. Optionally, after a period of rest, the compound can be administered under the same or a different schedule. The period of rest can be one, two, three, or four weeks, or longer, according to the pharmacodynamic effects of the compound on the subject.
[0089] The compound of the invention can be delivered to the subject by any suitable route, e.g., oral, rectal, buccal (e.g., sub-lingual), vaginal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), topical (i.e., both skin and mucosal surfaces, including airway surfaces) and transdermal administration. The compound is delivered to the subject at a dose that is effective to treat the disorder. The effective dosage will depend on many factors including the gender, age, weight, and general physical condition of the subject, the severity of the disorder, the particular compound or composition being administered, the duration of the treatment, the nature of any concurrent treatment, the carrier used, and like factors within the knowledge and expertise of those skilled in the art. As appropriate, a treatment effective amount in any individual case can be determined by one of ordinary skill in the art by reference to the pertinent texts and literature and/or by using routine experimentation (see, e.g., Remington, The Science and Practice of Pharmacy (21st ed. 2005)). In one embodiment, the compound is administered at a dose of about 0.001 to about 10 mg/kg body weight, e.g., about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mg/kg. In some instances, the dose can be even lower, e.g., as low as 0.0005 or 0.0001 mg/kg or lower. In some instances, the dose can be even higher, e.g., as high as 20, 50, 100, 500, or 1000 mg/kg or higher. The present invention encompasses every sub-range within the cited ranges and amounts.
[0090] In one aspect of the invention, the compound of the invention is delivered to a subject concurrently with an additional therapeutic agent. The additional therapeutic agent can be delivered in the same composition as the compound or in a separate composition. The additional therapeutic agent can be delivered to the subject on a different schedule or by a different route as compared to the compound. The additional therapeutic agent can be any agent that provides a benefit to the subject. Further agents include, without limitation, anti-psychotics, antidepressants, agents for neurological disorders, and chemotherapeutic agents.
[0091] Examples of anti-psychotic agents include, without limitation, for psychosis: prochloroperazine, chlorpromazine, droperidol, fluphenazine, periciazine, perphenazine, thiothixene, triflupromazine, haloperidol, molindone, pimozide, and thioridazine: and for schizophrenia: olanzapine, paliperidone, aripiprazole, quetiapine, risperidone, clozapine, flupenthixene, iloperidone, loxapine, mesoridazine, promazine, reserpine, thioridazine, zuclopenthixol, asenapine, levomepromazine, ziprasidone, molindone, pimozide, and thioridazine.
[0092] Examples of agents for depression and related disorders include, without limitation, for depression: modafinil, desvenlafaxine, nortriptyline, seleginline, amoxapine, citalopram, clomipramine, fluoxetine, isocarboxazid, mapriotiline, nefazodone, niacin, nortriptyline, phenelzine, protriptyline, tranylcypromine, trazodone, trimipramine, desipramine, imipramine, methylphenidate, buproprion, alprazolam, amitriptyline, chlordiazepoxide, perphenazine, doxepin, mirtazapine, 5-hydroxytryptophan, duloxetine, escitalopram, venlafaxine, desvenlafaxine, paroxetine, fluoxetine, olanzapine, 1-methylfolate, amitriptyline, sertraline, fluvoxamine, paliperidone, aripiprazole, quetiapine, risperidone, amisulpride; for anxiety: atomoxetine, aspirin, meprobamate, atenolol, buspirone, chlorodiazepoxide, chlorprothixene, clorazepate, diazepam, gabapentin, halazepam, hydroxyzine, lorazepam, meprobamate, nadolol, oxazepam, phenytoin, trifluoperazine, clonidine, clonazepam, oxcarbazepine, prochloroperazine, alprazolam, amitriptyline, chlordiazepoxide, perphenazine, doxepin, mirtazapine, 5- hydroxytryptophan, duloxetine, escitalopram, venlafaxine, desvenlafaxine, paroxetine, sertraline, fluvoxamine, amisulpride; and for bipolar disorder: clonidine, clonazapam, oxcarbazepine, carbamazepine, divalproex, lamotrigine, levetiracetam, lithium, topiramate, valproic acid, verapamil, buproprion, duloxetine, escitalopram, venlafaxine, 1-methylfolate, fluoxetine, olanzapine, sertraline, olanzapine, paliperidone, aripiprazole, quetiapine, risperidone, asenapine, levomepromazine, ziprasidone.
[0093] Examples of agents for neurological disorders include, without limitation, for Alzheimer's disease: caprylidene, donepezil, galantamine, tacrine, vitamin E, ergoloid mesylates, rivastigmine; for Parkinson's disease: nadolol, zonisamide, amantadine, apomorphine, belladonna, benztropine, biperiden, bromocriptine, carbidopa, entacapone, levodopa, pergolide mesylate, pramipexole, procyclidine, rasagiline, ropinirole, rotiotine, scopolamine, tolcapone, trihexylphenidyl, rivastigmine, seleginline; for Huntington's disease: baclofen, pregabalin, tetrabenazine, methylprednisolone, desvenlafaxine, nortriptyline; and for dementia: haloperidol and ergoloid mesylates.
[0094] Examples of chemotherapeutic agents include, without limitation, acivicin, aclarubicin, acodazole hydrochloride, acronine, adozelesin, aldesleukin, altretamine, ambomycin, ametantrone acetate, aminoglutethimide, amsacrine, anastrozole, anthramycin, asparaginase, asperlin, azacytidine, azetepa, azotomycin, batimastat, benzodepa, bicalutamide, bisantrene hydrochloride, bisnafide dimesylate, bizelesin, bleomycin sulfate, brequinar sodium, bropirimine, busulfan, cactinomycin, calusterone, caracemide, carbetimer, carboplatin, carmustine, carubicin hydrochloride, carzelesin, cedefingol, chlorambucil, cirolemycin, cisplatin, cladribine, crisnatol mesylate, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin hydrochloride, decitabine, dexormaplatin, dezaguanine, dezaguanine mesylate, diaziquone, docetaxel, doxorubicin, doxorubicin hydrochloride, droloxifene, droloxifene citrate, dromostanolone propionate, duazomycin, edatrexate, eflornithine hydrochloride, elsamitrucin, enloplatin, enpromate, epipropidine, epirubicin hydrochloride, erbulozole, esorubicin hydrochloride, estramustine, estramustine phosphate sodium, etanidazole, etoposide, etoposide phosphate, etoprine, fadrozole hydrochloride, fazarabine, fenretinide, floxuridine, fludarabine phosphate, fluorouracil, flurocitabine, fosquidone, fostriecin sodium, gemcitabine, gemcitabine hydrochloride, hydroxyurea, idarubicin hydrochloride, ifosfarnide, ilmofosine, interleukin II (including recombinant interleukin II or rIL2), interferon alfa-2a, interferon alfa-2b, interferon alfa-nl, interferon alfa-n3, interferon beta-la, interferon gamma-lb, iproplatin, irinotecan hydrochloride, lanreotide acetate, letrozole, leuprolide acetate, liarozole hydrochloride, lometrexol sodium, lomustine, losoxantrone hydrochloride, masoprocol, maytansine, mechlorethamine hydrochloride, megestrol acetate, melengestrol acetate, melphalan, menogaril, mercaptopurine, methotrexate, methotrexate sodium, metoprine, meturedepa, mitindomide, mitocarcin, mitocromin, mitogillin, mitomalcin, mitomycin, mitosper, mitotane, mitoxantrone hydrochloride, mycophenolic acid, nocodazole, nogalamycin, ormaplatin, oxisuran, paclitaxel, pegaspargase, peliomycin, pentamustine, peplomycin sulfate, perfosfamide, pipobroman, piposulfan, piroxantrone hydrochloride, plicamycin, plomestane, porfimer sodium, porfiromycin, prednimustine, procarbazine hydrochloride, puromycin, puromycin hydrochloride, pyrazofurin, riboprine, rogletimide, safingol, safingol hydrochloride, semustine, simtrazene, sparfosate sodium, sparsomycin, spirogermanium hydrochloride, spiromustine, spiroplatin, streptonigrin, streptozotocin, sulofenur, talisomycin, tecogalan sodium, tegafur, teloxantrone hydrochloride, temoporfin, teniposide, teroxirone, testolactone, thiamiprine, thioguanine, thiotepa, tiazofurin, tirapazamine, toremifene citrate, trestolone acetate, triciribine phosphate, trimetrexate, trimetrexate glucuronate, triptorelin, tubulozole hydrochloride, uracil mustard, uredepa, vapreotide, verteporfin, vinblastine sulfate, vincristine sulfate, vindesine, vindesine sulfate, vinepidine sulfate, vinglycinate sulfate, vinleurosine sulfate, vinorelbine tartrate, vinrosidine sulfate, vinzolidine sulfate, vorozole, zeniplatin, zinostatin, zorubicin hydrochloride.
[0095] Examples of other chemotherapeutic agents include, but are not limited to, 20-epi- 1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein- 1; prostatic carcinoma antiandrogen; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel; docosanol; dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-1 receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; 4- ipomeanol; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor 1- based therapy; mustard anticancer agent; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn; 06-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; odansteron; oracin; oral cytokine inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues; paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; prednisone; propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin; ribozymes; RJI retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone Bl; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine; senescence derived inhibitor 1 ; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen binding protein; sizofiran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B; vector system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.
[0096] The present invention finds use in research as well as veterinary and medical applications. Suitable subjects are generally mammalian subjects. The term "mammal" as used herein includes, but is not limited to, humans, non-human primates, cattle, sheep, goats, pigs, horses, cats, dog, rabbits, rodents (e.g., rats or mice), etc. Human subjects include neonates, infants, juveniles, adults and geriatric subjects.
[0097] In particular embodiments, the subject is a human subject that has a CNS disorder associated with D2 dopamine receptors. In other embodiments, the subject used in the methods of the invention is an animal model of a CNS disorder associated with D2 dopamine receptors.
[0098] The subject can be a subject "in need of the methods of the present invention, e.g., in need of the therapeutic effects of the inventive methods. For example, the subject can be a subject that is experiencing a CNS disorder associated with D2 dopamine receptors and/or is anticipated to experience a CNS disorder associated with D2 dopamine receptors, and the methods and compositions of the invention are used for therapeutic and/or prophylactic treatment.
[0099] The compounds of the invention described above can be formulated for administration in a pharmaceutical carrier in accordance with known techniques. See, e.g., Remington, The Science And Practice of Pharmacy (21st ed. 2005). In the manufacture of a pharmaceutical formulation according to the invention, the compound is typically admixed with, inter alia, an acceptable carrier. The carrier must, of course, be acceptable in the sense of being compatible with any other ingredients in the formulation and must not be deleterious to the patient. The carrier can be a solid or a liquid, or both, and can be formulated with the compound as a unit-dose formulation, for example, a tablet, which can contain from 0.01% or 0.5% to 95% or 99% by weight of the compound. One or more compounds can be incorporated in the formulations of the invention, which can be prepared by any of the well known techniques of pharmacy comprising admixing the components, optionally including one or more accessory ingredients.
[0100] The formulations of the invention include those suitable for oral, rectal, topical, buccal (e.g., sub-lingual), vaginal, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), topical (i.e., both skin and mucosal surfaces, including airway surfaces) and transdermal administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated and on the nature of the particular active compound which is being used.
[0101] Formulations suitable for oral administration can be presented in discrete units, such as capsules, cachets, lozenges, or tablets, each containing a predetermined amount of the active compound; as a powder or granules; as a solution or a suspension in an aqueous or nonaqueous liquid; or as an oil-in-water or water-in-oil emulsion. Such formulations can be prepared by any suitable method of pharmacy which includes the step of bringing into association the compound and a suitable carrier (which can contain one or more accessory ingredients as noted above). In general, the formulations of the invention are prepared by uniformly and intimately admixing the compound with a liquid or finely divided solid carrier, or both, and then, if necessary, shaping the resulting mixture. For example, a tablet can be prepared by compressing or molding a powder or granules containing the compound, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing, in a suitable machine, the compound in a free-flowing form, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent, and/or surface active/dispersing agent(s). Molded tablets can be made by molding, in a suitable machine, the powdered compound moistened with an inert liquid binder.
[0102] Formulations suitable for buccal (sub-lingual) administration include lozenges comprising the compound in a flavored base, usually sucrose and acacia or tragacanth; and pastilles comprising the compound in an inert base such as gelatin and glycerin or sucrose and acacia.
[0103] Formulations of the present invention suitable for parenteral administration comprise sterile aqueous and non-aqueous injection solutions of the compound, which preparations are preferably isotonic with the blood of the intended recipient. These preparations can contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation isotonic with the blood of the intended recipient. Aqueous and non-aqueous sterile suspensions can include suspending agents and thickening agents. The formulations can be presented in unit/dose (e.g., in a syringe or other injection device) or multi-dose containers, for example sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, saline or water-for-injection immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules and tablets of the kind previously described. For example, in one aspect of the present invention, there is provided an injectable, stable, sterile composition comprising one or more compounds, in a unit dosage form in a sealed container. The compound is provided in the form of a lyophilizate which is capable of being reconstituted with a suitable pharmaceutically acceptable carrier to form a liquid composition suitable for injection thereof into a subject. The unit dosage form typically comprises from about 0.001 mg to about 10 grams of the compound. When the compound is substantially water-insoluble (e.g., when conjugated to a lipid), a sufficient amount of emulsifying agent which is physiologically acceptable can be employed in sufficient quantity to emulsify the compound in an aqueous carrier. One such useful emulsifying agent is phosphatidyl choline.
[0104] Formulations suitable for rectal administration are preferably presented as unit dose suppositories. These can be prepared by admixing the compound with one or more conventional solid carriers, for example, cocoa butter, and then shaping the resulting mixture.
[0105] Formulations suitable for topical application to the skin preferably take the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil. Carriers which can be used include petroleum jelly, lanoline, polyethylene glycols, alcohols, transdermal enhancers, and combinations of two or more thereof.
[0106] Formulations suitable for transdermal administration can be presented as discrete patches adapted to remain in intimate contact with the epidermis of the recipient for a prolonged period of time. Formulations suitable for transdermal administration can also be delivered by iontophoresis (see, for example, Pharm. Res. 3:318 (1986)) and typically take the form of an optionally buffered aqueous solution of the compound. Suitable formulations comprise citrate or bis\tris buffer (pH 6) or ethanol/water and contain from 0.1 to 0.2 M active ingredient.
[0107] Other pharmaceutical compositions can be prepared from the compounds disclosed herein, such as aqueous base emulsions. In such an instance, the composition will contain a sufficient amount of pharmaceutically acceptable emulsifying agent to emulsify the desired amount of the compound. Particularly useful emulsifying agents include phosphatidyl cholines and lecithin.
[0108] In addition to compound, the pharmaceutical compositions can contain other additives, such as pH-adjusting additives. In particular, useful pH-adjusting agents include acids, such as hydrochloric acid, bases or buffers, such as sodium lactate, sodium acetate, sodium phosphate, sodium citrate, sodium borate, or sodium gluconate. Further, the compositions can contain microbial preservatives. Useful microbial preservatives include methylparaben, propylparaben, and benzyl alcohol. The microbial preservative is typically employed when the formulation is placed in a vial designed for multidose use. Other additives that are well known in the art include, e.g., detackifiers, anti-foaming agents, antioxidants (e.g., ascorbyl palmitate, butyl hydroxy anisole (BHA), butyl hydroxy toluene (BHT) and tocopherols, e.g., a-tocopherol (vitamin E)), preservatives, chelating agents (e.g., EDTA and/or EGTA), viscomodulators, tonicifiers (e.g., a sugar such as sucrose, lactose, and/or mannitol), flavorants, colorants, odorants, opacifiers, suspending agents, binders, fillers, plasticizers, lubricants, and mixtures thereof. The amounts of such additives can be readily determined by one skilled in the art, according to the particular properties desired.
[0109] The additive can also comprise a thickening agent. Suitable thickening agents can be those known and employed in the art, including, e.g., pharmaceutically acceptable polymeric materials and inorganic thickening agents. Exemplary thickening agents for use in the present pharmaceutical compositions include polyacrylate and polyacrylate co-polymer resins, for example poly-acrylic acid and poly-acrylic acid/methacrylic acid resins; celluloses and cellulose derivatives including: alkyl celluloses, e.g., methyl-, ethyl- and propyl-celluloses; hydroxyalkyl- celluloses, e.g., hydroxypropyl-celluloses and hydroxypropylalkyl-celluloses such as hydroxypropyl-methyl-celluloses; acylated celluloses, e.g., cellulose-acetates, cellulose- acetatephthallates, cellulose-acetatesuccinates and hydroxypropylmethyl-cellulose phthallates; and salts thereof such as sodium-carboxymethyl-celluloses; polyvinylpyrrolidones, including for example poly-N-vinylpyrrolidones and vinylpyrrolidone co-polymers such as vinylpyrrolidone- vinylacetate co-polymers; polyvinyl resins, e.g., including polyvinylacetates and alcohols, as well as other polymeric materials including gum traganth, gum arabicum, alginates, e.g., alginic acid, and salts thereof, e.g., sodium alginates; and inorganic thickening agents such as atapulgite, bentonite and silicates including hydrophilic silicon dioxide products, e.g., alkylated (for example methylated) silica gels, in particular colloidal silicon dioxide products. Such thickening agents as described above can be included, e.g., to provide a sustained release effect. However, where oral administration is intended, the use of thickening agents as aforesaid will generally not be required and is generally less preferred. Use of thickening agents is, on the other hand, indicated, e.g., where topical application is foreseen.
[0110] Further, the present invention provides liposomal formulations of the compounds disclosed herein. The technology for forming liposomal suspensions is well known in the art. When the compound is in the form of an aqueous-soluble material, using conventional liposome technology, the same can be incorporated into lipid vesicles. In such an instance, due to the water solubility of the compound, the compound will be substantially entrained within the hydrophilic center or core of the liposomes. The lipid layer employed can be of any conventional composition and can either contain cholesterol or can be cholesterol-free. When the compound of interest is water-insoluble, again employing conventional liposome formation technology, the compound can be substantially entrained within the hydrophobic lipid bilayer which forms the structure of the liposome. In either instance, the liposomes which are produced can be reduced in size, as through the use of standard sonication and homogenization techniques. The liposomal formulations containing the compound disclosed herein, can be lyophilized to produce a lyophilizate which can be reconstituted with a pharmaceutically acceptable carrier, such as water, to regenerate a liposomal suspension.
[0111] The present invention is explained in greater detail in the following non-limiting Examples.
EXAMPLE 1
Synthesis of Compounds
[0112] The compounds shown in Table 5 were synthesized as described below.
Figure imgf000040_0001
Figure imgf000041_0001
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
General Procedures.
[0113] Unless stated to the contrary, where applicable, the following conditions apply: all commercial grade reagents were used without further purification. MeCN and CH2C12 were distilled from CaH2 under a N2 atmosphere prior to use; THF was distilled from Na/benzophenone under N2. All other dry solvents were of anhydrous quality purchased from Sigma-Aldrich. Brine (NaCl), NaHC03, and NH4C1 refer to saturated aqueous solutions. Melting points were uncorrected. Mass spectra (MS) and HPLC (UV 254 nM or ELSD) data for all compounds were recorded on an 1100 LC/MS system (Agilent Technology Corporation) using a 4.6x50 mm column (CenturySIL C-18 AQ+, 5μ) with a linear gradient 30-90% (v/v) acetonitrile-water with 0.035% trifluoroacetic acid over 5 min with a flow rate of 3.5 mL/min. Analytical TLC was performed using 2.5 x 5 cm plates coated with 0.25 mm of silica gel GF254. Column chromatography was performed on silica gel G (200-300 mesh) with reagent grade solvents. ¾ NMR spectra (300 MHz) are reported as follows: chemical shifts in ppm (δ scale) downfield from TMS as the internal standard with either CDC13 or DMSO-i/6 as the solvent. Multiplicities are indicated as the following: bs=broad singlet, s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet.
Synthesis of Compound 1
Scheme 1. Synthesis of intermediate 2
Scheme 2. Synthesis of intermediate 3
Figure imgf000045_0002
Intermediate 3
Scheme 3. Synthesis of compound 1
Figure imgf000045_0003
Example 1
Intermediate 1:
Figure imgf000045_0004
[0114] A well-dried flask was first charged with l-bromo-2,3-dichlorobenzene (4 g, 17.7 mmol) and 1,4-diazepane (2.1 g, 20.9 mmol), which was evacuated and backfilled with N2 through a balloon under gentle warming (40°C). Toluene was charged and the mixture was bubbled with N2 for 10 min, then BINAP (318 mg, 0.51 mmol) and Pd2dba3 (156 mg, 0.17 mmol) was added to the mixture, followed by DBU (3.4 mL). The resulting mixture was warmed at 60- 70°C while fine powder of tBuONa was added in one portion to start the amination. After the reaction mixture cooled to rt, (Boc)20 (11 g, 51 mmol) solution in DCM was added dropwise to the reaction mixture, then stirred for 3 h at rt. The reaction mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 10:1) to give tert-butyl 4-(2,3-dichlorophenyl)-l,4-diazepane- 1-carboxylate (intermediate 1) (4.1 g, 70%) as a yellow oil.
Intermediate 2:
Figure imgf000046_0001
[0115] Excess HC1 in EtOAc was added dropwise to a solution of intermediate 1 (4.0 g, 11.6 mmol) in EtOAc and the reaction mixture was stirred at rt for 1.5 h. The reaction mixture was filtered to give l-(2,3-dichlorophenyl)-l,4-diazepane hydrochloride salt (intermediate 2) (3.0 g, 90%) as a white solid. HPLC: 99%, RT 2.108 min. MS (ESI) m/z 245.0 [M + H]+. mp: 186- 187°C.
Intermediate 3:
Figure imgf000046_0002
[0116] 7-hydroxy-3,4-dihydroquinolin-2(lH)-one (195 mg, 1.2 mmol), 1,3- dibromopropane (960 mg, 4.8 mmol) and anhydrous K2C03 (166 mg, 1.2 mmol) were dissolved in EtOH and the solution was heated to reflux overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 2:1) to give 7-(3-bromopropoxy)-3,4- dihydroquinolin-2(lH)-one (intermediate 3) (300 mg, 88%) as a white solid. HPLC: 99%, RT 2.775 min. MS (ESI) m/z 284.0 [M + H]+.
Compound 1:
Figure imgf000046_0003
[0117] A mixture of intermediate 3 (142 mg, 0.5 mmol) and Nal (150 mg, 1.0 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (196 mg, 0.7 mmol) and anhydrous K2C03 (173 mg, 1.25 mmol) were added to the mixture. The resulting mixture was heated to reflux for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 7-(3-(4- (2,3-dichlorophenyl)-l,4-diazepan-l-yl)propoxy)-3,4-dihydroquinolin-2(lH)-one (compound 1) (140 mg, 63%). ¾ NMR (300 MHz, CDC13): δ 8.16 (s, 1H), 7.10-6.98 (m, 4H), 6.53 (dd, J = 8.1 Hz, 2.4 Hz, 1H), 6.34 (d, J = 2.1 Hz, 1H), 4.01 (t, J = 6.6 Hz, 2H), 3.32-3.28 (m, 4H), 2.92-2.87 (m, 6H), 2.76 (t, J = 6.6 Hz, 2H), 2.62 (t, J = 6.6 Hz, 2H), 2.04-1.97 (m, 4H). HPLC: 99%, RT 2.421 min. MS (ESI) m/z 448.1 [M + H]+. mp: 130-131°C.
Synthesis of Compound 2
Scheme 4. Synthesis of intermediate 4
Figure imgf000047_0001
Intermediate 4
Scheme 5. Synthesis of compound 2
Figure imgf000047_0002
Intermediate 4:
Figure imgf000047_0003
[0118] 7-hydroxy-3,4-dihydroquinolin-2(lH)-one (163 mg, 1.0 mmol), 1,4- dibromobutane (0.36 mL, 3.0 mmol) and anhydrous K2C03 (138 mg, 1.0 mmol) were dissolved in EtOH and the solution was heated to reflux overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE EtOAc = 2: 1) to give 7-(4-bromobutoxy)-3,4-dihydroquinolin- 2(lH)-one (intermediate 4) (220 mg, 74%) as a white solid, mp: 106-109°C. Compound 2:
Figure imgf000048_0001
[0119] A mixture of intermediate 4 (156 mg, 0.52 mmol) and Nal (120 mg, 0.8 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (119 mg, 0.4 mmol) and anhydrous K2C03 (189 mg, 1.37 mmol) were then added to the mixture. The resulting mixture was heated to reflux for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2,3-dichlorophenyl)-l ,4-diazepan-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one
(compound 2) (75 mg, 41%). ¾ NMR (300 MHz, CDC13): δ 8.08 (s, 1H), 7.10-6.99 (m, 4H), 6.52 (dd, J = 6.6 Hz, 2.1 Hz, 1H), 6.33 (s, 1H), 3.96 (t, J= 6.0 Hz, 2H), 3.31 -3.27 (m, 4H), 2.92- 2.84 (m, 6H), 2.64-2.60 (m, 4H), 2.02-1.98 (m, 2H), 1.84-1.70 (m, 4H). HPLC: 99%, RT 2.435 min. MS (ESI) m/z 462.2 [M + H]+. mp: 88-89°C.
Synthesis of Compound 3
Scheme 6. Synthesis of intermediate 12
Figure imgf000048_0002
intermediate 9 intermediate 10
Figure imgf000048_0003
ntermediate 11 intermediate 12 Scheme 7. Synthesis of compound 3
Figure imgf000049_0001
[0120] Pivaloyl chloride (18.7 mL, 151.6 mmol) was slowly added to an ice-cooled solution of 6-chloropyridin-2-amine (15 g, 116.7 mmol) and triethylamine (24.25 mL, 124.96 mmol) in DCM (180 mL). The mixture was stirred in an ice-bath for 15 min and then at rt for 6 h. The mixture was poured into water. The organic layer was washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04; filtered and concentrated in vacuo to give N-(6- chloropyridin-2-yl)pivalamide (intermediate 5) (20.01 g, 81%) as a yellow solid.
Intermediate 6:
Figure imgf000049_0002
[0121] A solution of intermediate 5 (19.28 g, 90.6 mmol) in THF (181 mL) was treated with 77-BuLi (108.8 mL, 272 mmol) and the resulting mixture was stirred at -20°C for 3 h. After addition of DMF (20.81 mL, 271.86 mmol), the reaction was allowed to warm to rt. The reaction was poured into cold 6N HC1 and stirred for 15 min. The mixture was then neutralized with anhydrous K2C03 to pH=7 and extracted with Et20. The combined organic layers were washed with water, brine, dried over anhydrous Na2SC>4, concentrated in vacuo and purified by recrystallization from EtOAc and hexanes to afford N-(6-chloro-3-formylpyridin-2-yl)pivalamide (intermediate 6) (12.84 g, 59%).
Intermediate 7:
Figure imgf000049_0003
[0122] To a solution of 4-(benzyloxy)butan-l-ol (15.42 g, 85.6 mmol) in DMF (120 mL) was added sodium hydride (4.11 g, 171 mmol) at 0°C. The mixture was stirred for 20 min, then intermediate 6 (10.28 g, 42.7 mmol) was added portion-wise and the resulting mixture was stirred overnight. The mixture was quenched with saturated aq NH CI and extracted with EtOAc. The combined organic layers were washed with water, brine, dried over anhydrous Na2S04 and concentrated in vacuo. The residue was purified by flash chromatography on silica gel column (elution with PE EtOAc = 8: 1 - 4: 1) to give N-(6-(4-(benzyloxy)butoxy)-3-formylpyridin-2- yl)pivalamide (intermediate 7) (5.04 g, 31%).
Intermediate 8:
Figure imgf000050_0001
[0123] Under N2, an ice-cooled flask containing THF (100 mL) was charged with sodium hydride (5.7 g, 142.5 mmol) to which ethyl 2-(diethoxyphosphoryl)acetate (32 g, 142 mmol) was added. The ice bath was removed and a solution of intermediate 7 (21.8 g, 56.7 mmol) in THF (120 mL) was slowly added to the preformed anion. The reaction was heated to reflux overnight. The reaction mixture was cooled to rt and partitioned between EtOAc and water. The organic layer was separated and the aqueous layer was extracted with EtOAc. The combined organic layers were washed with brine, dried over anhydrous Na2S04 and concentrated in vacuo to give ethyl 3-(6-(4-(benzyloxy)butoxy)-2-pivalamidopyridin-3-yl)acrylate (intermediate 8) (13.3 g, 52%) as a white solid, mp: 75-76°C.
Intermediate 9:
Figure imgf000050_0002
[0124] Intermediate 8 (2.46 g, 5.4 mmol) was hydrogenated under an atmosphere of ¾ (40 psi) using Raney Ni (1.59 g, 27.1 mmol) in THF (120 mL). The reaction was filtered to remove the Raney Ni and the filtrate was concentrated to afford ethyl 3-(6-(4- (benzyloxy)butoxy)-2-pivalamidopyridin-3-yl)propanoate (intermediate 9) (1.76 g, 71.5% crude) as an oil.
Intermediate 10:
Figure imgf000050_0003
[0125] To a solution of intermediate 9 (1.76 g, 3.9 mmol) in 1,4-dioxane (14 mL) was added 3N HC1 (7.1 mL) and the mixture was then heated to reflux for 3 h. The mixture was cooled to rt and then anhydrous K2C03 was slowly added to neutralize the reaction mixture to pH=8, then extracted with EtOAc. The combined organic layers were washed with water, brine, dried over anhydrous Na2S04 and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel column (elution with PE EtOAc = 4: 1 - 1 : 1) to give 7-(4- (benzyloxy)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one (intermediate 10) (671 mg, 54%) as an oil. Intermediate 11:
Figure imgf000051_0001
[0126] To the solution of intermediate 10 (900 mg, 2.8 mmol) in EtOH (25 mL) was added 10% Pd/C (400 mg) under an atmosphere of H2 overnight. The reaction was filtered to remove the Pd/C and the filtrate was concentrated and washed with Et20 to give 7-(4- hydroxybutoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one (intermediate 11) (360 mg, 55%) as a white solid.
Intermediate 12:
Figure imgf000051_0002
[0127] A mixture of intermediate 11 (140 mg, 0.593 mmol), Et3N (240 mg, 2.37 mmol) and methylsulfonyl chloride (102 mg, 0.89 mmol) was dissolved in DCM and the solution was stirred for 6 h at rt. The solution was then washed with water, brine, and dried over anhydrous Na2S04. After filtration, the solvent was evaporated in vacuo to give 4-(7-oxo-5,6,7,8- tetrahydro-l,8-naphthyridin-2-yloxy)butyl methanesulfonate (intermediate 12) (180 mg, 96%) as a yellow solid.
Compound 3:
Figure imgf000051_0003
[0128] Intermediate 12 (85 mg, 0.27 mmol) was dissolved in CH3CN then Nal (55 mg, 0.37 mmol), Et3N (91 mg, 0.9 mmol), and intermediate 2 (102 mg, 0.36 mmol) were added. The resulting mixture was stirred at 70°C overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous a2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8- naphthyridin-2(lH)-one (compound 3) (78 mg, 63%). ¾ MR (300 MHz, CDC13): δ 7.64 (s, 1H), 7.38 (d, J = 8.1 Hz, 1H), 7.27-7.15 (m, 2H), 7.06 (d, J = 8.1 Hz, 1H), 6.37 (d, J = 8.1 Hz, 1H), 4.36 (t, J- 6.0 Hz, 2H), 3.61-3.49 (m, 7H), 3.32-3.20 (m, 5H), 2.90-2.85 (m, 2H), 2.67-2.62 (m, 2H), 2.21-2.16 (m, 2H), 1.90-1.85 (m, 2H). HPLC: 99%, RT 2.682 min. MS (ESI) m/z 463.1 [M + H]+. mp: 110-111°C. Synthesis of Compound 4
Scheme 8. Synthesis of intermediate 15
Figure imgf000052_0001
Intermediate 13:
Figure imgf000052_0002
[0129] A solution of cinnamoyl chloride (4.25 g, 25.6 mmol) in acetone (25 mL) was added to a solution of 3-methoxyaniline (2.74 mL, 24.4 mmol) and anhydrous K2CO3 (3.7 g, 26.8 mmol) in acetone-ice (28 mL-28 g) and the resulting reaction mixture was stirred at rt for 3 h. The reaction mixture was then poured into ice water, filtered, and washed with water. Recrystallization from EtOH gave N-(3-methoxyphenyl)cinnamamide (intermediate 13) (4.56 g, 74%) as a white solid.
Intermediate 14:
Figure imgf000052_0003
[0130] AICI3 (11.5 g, 86.3 mmol) was added portion-wise to a suspension of intermediate 13 (4.2 g, 16.6 mmol) in chlorobenzene (90 mL) at 0°C. The reaction mixture was gradually warmed to 120°C and stirred for 3 h. The mixture was poured into ice water and the resulting precipitate was collected by filtration, washed with water, and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 60:1) to give 7-hydroxyquinolin-2(lH)-one (intermediate 14) (1.41 g, 53%) as a white solid. Intermediate 15:
Figure imgf000053_0001
[0131] A mixture of intermediate 14 (193 mg, 1.2 mmol), 1,4-dibromobutane (0.43 mL, 3.6 mmol) and anhydrous K2C03 (166 mg, 1.2 mmol) was dissolved in EtOH and the solution was heated to reflux overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers was washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 2:1) to give 7-(4-bromobutoxy)quinolin-2(lH)-one (intermediate 15) (147 mg, 41%) as a yellow solid.
Compound 4:
Figure imgf000053_0002
[0132] A mixture of intermediate 15 (120 mg, 0.41 mmol) and al (123 mg, 0.82 mmol) in C¾CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (175 mg, 0.62 mmol) and anhydrous K2C03 (226 mg, 1.64 mmol) were added to the mixture. The resulting mixture was heated to reflux overnight. The mixture was evaporated under reduced pressure and the residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2,3-dichlorophenyl)-l,4- diazepan-l-yl)butoxy)quinolin-2(lH)-one (compound 4) (50 mg, 26%). Ή NMR (300 MHz, CDC13) δ 11.54 (br„ 1H), 7.71 (d, J = 6.9 Hz, 1H) 7.44 (d, J = 8.1 Hz, 1H), 7.09-7.07 (m, 2H), 7.01-6.97 (m, 1H), 6.82-6.78 (m, 2H), 6.53 (d, J= 9.3 Hz, 1H), 4.10 (t, J= 6.6 Hz, 2H), 3.30 (d, J = 5.1 Hz, 4H), 2.87-2.81 (m, 4H), 2.62 (t, J= 7.5 Hz, 2H), 2.01-1.99 (m, 2H), 1.92-1.82 (m, 2H), 1.76-1.68 (m, 2H). HPLC: 99%, RT 2.461 min. MS (ESI) m/z 460.2 [M + H]+. mp: 55-57°C.
Synthesis of Compound 5
Figure imgf000054_0001
BF3/MeOH MsCl then TiCI4
Figure imgf000054_0002
Scheme 11. Synthesis of compound 5
Figure imgf000054_0003
Example 5
Intermediate 16:
Figure imgf000054_0004
[0133] To the solution of 6-methoxy-2,3-dihydro-lH-inden-l-one (6.5 g, 40 mmol) in 100 mL of MeOH was added Et3N (11.1 mL, 80 mmol) and hydroxylamine hydrochloride (3.9 g, 56 mmol). The mixture was stirred at rt for 2 h. The mixture was evaporated and redissolved in EtOAc, the organic phase was washed with water, brine, dried over anhydrous a2S04, filtered and concentrated to give (£)-6-methoxy-2,3-dihydro-lH-inden-l-one oxime (intermediate 16) (6.6 g, 93%).
Intermediate 17:
Figure imgf000054_0005
[0134] To the solution of intermediate 16 (6.6 g, 37.2 mmol) in CH2C12 (120 mL) was added Et3N (20.6 mL, 149 mmol), DMAP (455 mg, 3.73 mmol) and MsCl (4.35 mL, 55.87 mmol). The mixture was stirred at rt overnight. The reaction mixture was washed successively with water, saturated aq NH4C1, brine, dried over anhydrous MgS04, filtered and concentrated. The crude product was purified by flash chromatography on silica gel column (elution with PE/CH2C12 = 1 : 1) to give (£)-6-methoxy-2,3-dihydro-lH-inden-l-one O-methylsulfonyl oxime (intermediate 17) (7.76 g, 82%) as a white solid.
Intermediate 18:
Figure imgf000055_0001
[0135] To the solution of intermediate 17 (7.76 g, 30.4 mmol) in 1,2-dichloroethane (150 mL) was added BF3 (5.52 mL, 48.7 mmol, in MeOH), MsCl (3.8 mL, 48.7 mmol) and TiCl4 (5.35 mL, 48.7 mmol). The mixture was stirred at rt for 5 h. CH2C12 (150 mL) was then added to the reaction mixture. The solution was washed with water and brine. The water layer was neutralized by aqueous Na2C03 to pH = 7 and was extracted with CH2C12. The combined organic layers were dried over anhydrous MgS04, filtered and concentrated. The crude product was purified by flash chromatography on silica gel column (elution with PE EtOAc = 4: 1) to give 7- methoxy-3,4-dihydroisoquinolin-l(2H)-one (intermediate 18) (5.1 g, 95%) as a white solid.
Intermediate 19:
Figure imgf000055_0002
[0136] BBr3 (4.75 mL, 47.5 mmol, in CH2C12) was added to a cold solution (ice-bath) of intermediate 18 (3.36 g, 18.98 mmol) in CH2C12 (100 mL). The mixture was then warmed to rt and stirred overnight. Water was carefully added dropwise to quench the reaction. EtOAc was added and the mixture was washed with water, brine, dried over anhydrous MgS04, filtered and concentrated to afford 7-hydroxy-3,4-dihydroisoquinolin-l(2H)-one (intermediate 19) (2.46 g, 80%) as a white solid, mp: 201-203°C.
Intermediate 20:
Figure imgf000055_0003
[0137] A mixture of intermediate 19 (326 mg, 2 mmol), 1,3-dibromopropane (0.62 mL, 6 mmol) and anhydrous K2C03 (276 mg, 2 mmol) was dissolved in 4 mL of EtOH and the solution was heated to reflux and stirred overnight. The mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with water, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE EtOAc = 5:1) to give 7-(3-bromopropoxy)-3,4-dihydroisoquinolin-l(2H)-one (intermediate 20) (360 mg, 63%) as a white solid. Compound 5:
Figure imgf000056_0001
[0138] A mixture of intermediate 20 (110 mg, 0.39 mmol) and Nal (117 mg, 0.78 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (165 mg, 0.58 mmol) and anhydrous K2C03 (216 mg, 1.56 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)propoxy)-3,4-dihydroisoquinolin-l(2H)-one (compound 5) (125 mg, 71%). !H NMR (300 MHz, CDCI3) δ 7.60 (d, J = 2.1 Hz, 1H), 7.14-7.08 (m, 3H), 7.02-7.00 (m, 2H), 6.02 (br., 1H), 4.10 (t, J = 6.3 Hz, 2H), 3.56-3.53 (m, 2H), 3.31-3.27 (m, 4H), 2.96-2.88 (m, 6H), 2.77-2.72 (m, 2H), 2.01(m, 4H). HPLC: 99%, RT 2.578 min. MS (ESI) m/z 448.1 [M + H]+. mp: 116-117°C.
Synthesis of Compound 6
Scheme 12. Synthesis of intermediate 21
Figure imgf000056_0002
Scheme 13. Synthesis of compound 6
Figure imgf000056_0003
Intermediate 21:
Figure imgf000056_0004
[0139] A mixture of intermediate 19 (196 mg, 1.2 mmol), 1,4-dibromobutane (0.43 mL, 3.6 mmol) and anhydrous K2C03 (166 mg, 1.22 mmol) was dissolved in EtOH (4 mL) and the solution was heated to reflux and stirred overnight. The mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with water, brine, dried over anhydrous Na2SC>4, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 3 : 1) to give 7-(4-bromobutoxy)-3,4-dihydroisoquinolin-l(2H)- one (intermediate 21) (230 mg, 63%) as a white solid.
Compound 6:
Figure imgf000057_0001
[0140] A mixture of intermediate 21 (100 mg, 0.34 mmol) and Nal (102 mg, 0.68 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (144 mg, 0.51 mmol) and anhydrous K2CO3 (188 mg, 1.36 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous a2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 7-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)butoxy)-3,4-dihydroisoquinolin-l(2H)-one (compound 6) (107 mg, 68%). ¾ NMR (300 MHz, CDCI3) δ 7.58 (s, 1H), 7.20-7.09 (m, 3H), 7.02-6.98 (m, 2H), 6.04 (br„ 1H), 4.05 (t, J = 6.6 Hz, 2H), 3.54-3.52 (m, 2H), 3.32-3.28 (m, 4H), 2.95-2.91 (m, 6H), 2.70-2.68 (m, 2H), 2.06-2.04 (m, 2H), 1.84-1.77 (m, 4H). HPLC: 99%, RT 2.676 min. MS (ESI) m/z 462.4 [M + H]+. mp: 86-88°C.
Synthesis of Compound 7
Scheme 14. Synthesis of intermediate 26
Figure imgf000057_0002
intermediate 26 intermediate 25 intermediate 24 Scheme 15. Synthesis of compound 7
Figure imgf000058_0001
Intermediate 26
Intermediate 22:
Figure imgf000058_0002
[0141] A sample of 3-nitro-4-aminoanisole (13.5 g, 80 mmol) was dissolved in water (100 mL) and concentrated H2S04 (44 mL), cooled to 0°C, and diazotized with a solution of sodium nitrite (14.3 g, 207.2 mmol) in water (25 mL) while maintaining a temperature no greater than 5°C. The resulting solution was added dropwise with vigorous stirring to a solution of crystalline cobalt chloride (42 g, 176 mmol) and potassium thiocyanate (25.6 g, 264 mmol) in water (100 mL), after which the mixture was stirred at 0-10°C for 3 h and allowed to stand overnight. It was then stirred at 50°C for 16 h until nitrogen evolution had ceased. The precipitate was removed by filtration, washed with water, and dried at 80°C to give 4-methoxy-2- nitro-l-thiocyanatobenzene (intermediate 22) (9.82 g, 58%). mp: 127-128°C.
Intermediate 23:
Figure imgf000058_0003
[0142] A solution of intermediate 22 (4 g, 19 mmol) in benzene was added dropwise with stirring and cooling to a suspension of technical-grade lithium aluminum hydride (9.4 g, 247 mmol) in absolute ether (90 mL), after which the mixture was heated to reflux and stirred under a stream of dry nitrogen for 8 h. The excess lithium aluminum hydride was quenched by ice water. Water (25 mL) was then added, the mixture acidified to pH = 6 with dilute HC1, and then extracted with benzene. The extract was dried with MgS04, evaporated, and purified by flash column chromatography to afford 2-amino-4-methoxybenzenethiol hydrochloride (intermediate 23) (1.43 g, 49%) as a light yellow solid.
Intermediate 24:
Figure imgf000058_0004
[0143] A mixture of intermediate 23 (1.33 g, 6.9 mmol), boric acid (2.3 g, 37.2 mmol) and formic acid (17 mL) was heated to reflux and stirred for 6 h. The reaction mixture was cooled to rt and diluted with water. The solution was made alkaline and extracted with ethyl acetate. Further purification by column chromatography (elution with PE EtOAc = 3:1) gave 5- methoxybenzo[JJthiazole (intermediate 24) (290 mg, 15%). HPLC: 99%, RT 2.646 min. MS (ESI) m/z 166.0 [M + H]+.
Intermediate 25:
Figure imgf000059_0001
[0144] A mixture of intermediate 24 (272 mg, 1.65 mmol) and hydroiodic acid (45%, 2 mL) was heated to reflux for 5 h. The mixture was diluted with water and filtered. The filtrate was neutralized and the solids were filtered, washed with water, and combined to give benzo[c ]thiazol-5-ol (intermediate 25) (206 mg, 83%). HPLC: 99%, RT 2.038 min. MS (ESI) m/z 152.1 [M + H]+.
Intermediate 26:
Figure imgf000059_0002
[0145] A mixture of intermediate 25 (206 mg, 1.4 mmol), 1,4-dibromobutane (0.5 mL, 4.2 mmol) and anhydrous K2C03 (193 mg, 1.4 mmol) in EtOH (20 mL) was heated to reflux and stirred overnight. The yellow solid was filtered and purified by column chromatography (elution with PE/EtOAc = 4:1) to afford 5-(4-bromobutoxy)benzo[if]thiazole (intermediate 26) (108 mg, 39%) as a yellow oil. HPLC: 99%, RT 3.426 min. MS (ESI) m/z 288.0 [M + H]+.
Compound 7:
Figure imgf000059_0003
[0146] A mixture of intermediate 26 (47 mg, 0.16 mmol) and Nal (48 mg, 0.32 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (70 mg, 0.25 mmol) and anhydrous K2C03 (88 mg, 0.64 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 5-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)butoxy)benzo[if]thiazole (compound 7) (31 mg, 42%). ¾ NMR (300 MHz, CDC13) δ 8.97 (s, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.60 (d, J = 3.0 Hz, 1H), 7.11-7.07 (m, 3H), 7.01-6.98 (m, 1H), 4.10 (t, J= 5.7 Hz, 2H),), 3.33-3.27 (m, 4H), 2.96-2.95 (m, 4H), 2.73-2.71 (m, 2H), 2.06-2.04 (m, 2H), 1.92-1.80 (m, 4H), HPLC: 99%, RT 2.702 min. MS (ESI) m/z 450.1 [M + H]+. mp: 82- 84°C.
Synthesis of Compound 8
Scheme 16. Synthesis of intermediate 27
Figure imgf000060_0001
Scheme 17. Synthesis of compound 8
Figure imgf000060_0002
Intermediate 27
Intermediate 27:
Figure imgf000060_0003
[0147] A mixture of intermediate 25 (1 g, 6.6 mmol), 1,3-dibromopropane (1.83 mL, 18 mmol), anhydrous K2C03 (828 mg, 6.0 mmol) and EtOH (20 mL) was heated to reflux and stirred overnight. The yellow solid was filtered and purified by column chromatography (elution with PE/EtOAc = 3:1) to afford 5-(3-bromopropoxy)benzo[d]thiazole (intermediate 27) (543 mg, 33%) as a yellow oil.
Compound 8:
Figure imgf000060_0004
[0148] A mixture of intermediate 27 (106 mg, 0.39 mmol) and Nal (140 mg, 0.78 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (165 mg, 0.59 mmol) and anhydrous K2C03 (215 mg, 1.56 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 5-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)propoxy)benzo[<fJthiazole (compound 8) (98 mg, 60%) as a yellow solid. 'H NMR (300 MHz, DMSO-d6) δ 9.37 (s, 1H), 8.05 (d, J= 10.5 Hz, 1H), 7.66 (d, J= 4.8 Hz, 1H), 7.32-7.30 (m, 2H), 7.25-7.23 (m, 1H), 7.14 (dd, J = 9.0 Hz, 2.4 Hz, 1H), 4.19 (t, J = 6.0 Hz, 2H), 3.61-3.29 (m, 10H), 2.31-2.26 (m, 4H). HPLC: 99%, RT 2.601 min. MS (ESI) m/z 436.3 [M + H]+. mp: 93- 94°C.
Synthesis of Compound 9
Scheme 18. Synthesis of intermediate 29
Figure imgf000061_0001
Intermediate28 Intermediate 29
Scheme 19. Synthesis of compound 9
Figure imgf000061_0002
Intermediate 28:
Figure imgf000061_0003
[0149] A well-dried flask was first charged with l-bromo-2-methoxybenzene (2.5 g, 13.4 mmol) and 1,4-diazepane (1.6 g, 16.1 mmol), which was then evacuated and backfilled with N2 through a balloon under gentle warming (40°C). Toluene was added and the mixture was bubbled with N2 for 10 min, then BINAP (250 mg, 0.4 mmol) and Pd2dba3 (156 mg, 0.17 mmol) was added to the mixture. After the addition of DBU (2.4 mL), the solution was warmed at 60- 70°C while tBuONa (2.25 g, 20.1 mmol) was added in one portion to start the amination. After the reaction mixture cooled to rt, (Boc)20 (7.3 g, 33.5 mmol) was dissolved in DCM and added dropwise to the reaction mixture and stirred for 3 h at rt. The reaction mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 30:1) to give tert-butyl 4-(2-methoxyphenyl)- 1,4-diazepane- 1- carboxylate (intermediate 28) (396 mg, 10%) as a yellow oil. Intermediate 29:
Figure imgf000062_0001
[0150] Excess HC1 in EtOAc was added dropwise to a solution of intermediate 28 (396 mg, 1.29 mmol) in EtOAc and the reaction mixture was stirred at rt for 1.5 h. Filtration gave 1- (2-methoxyphenyl)-l,4-diazepane hydrochloride (intermediate 29) (200 mg, 64%) as a white solid.
Compound 9:
Figure imgf000062_0002
[0151] A mixture of intermediate 4 (150 mg, 0.51 mmol) and Nal (150 mg, 1.02 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 29 (121 mg, 0.5 mmol) and anhydrous K2C03 (276 mg, 2.0 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 7-(4-(4-(2-methoxyphenyl)-l,4-diazepan-l- yl)butoxy)-3,4-dihydroquinolin-2(lH)-one (compound 9) (25 mg, 11%). HPLC: 95%, RT 2.314 min. MS (ESI) m/z 424.2 [M + H]+.
Synthesis of Compound 10
Scheme 20. Synthesis of compound 10
Figure imgf000062_0003
Compound 10:
Figure imgf000062_0004
[0152] A mixture of intermediate 15 (150 mg, 0.51 mmol) and Nal (150 mg, 1.02 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 29 (124 mg, 0.51 mmol) and anhydrous K2C03 (282 mg, 2.04 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried Over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 7-(4-(4-(2-methoxyphenyl)-l,4-diazepan-l- yl)butoxy)quinolin-2(lH)-one (compound 10) (61 mg, 26%). *H NMR (300 MHz, CDC13) δ 7.69 (d, J = 9.6 Hz, 1H), 7.43 (d, J= 8.7 Hz, 1H), 6.91-6.85 (m, 5H), 6.79 (d, J= 9.0 Hz, 1H), 6.65 (s, 1H) 6.49 (d, J= 9.3 Hz, 1H), 4.07 (t, J = 6.0 Hz, 2H), 3.83 (s, 3H), 3.34-3.29 (m, 4H), 2.93-2.86 (m, 4H), 2.69-2.65 (m, 2H), 2.06-2.02 (m, 2H), 1.86-1.84 (m, 2H), 1.77-1.73 (m, 2H). HPLC: 95%, RT 1.902 min. MS (ESI) m/z 422.3 [M + H]+.
Synthesis of Compound 11
Scheme 21. Synthesis of compound 11
Figure imgf000063_0001
[0153] A mixture of intermediate 12 (150 mg, 0.48 mmol) and Nal (141 mg, 0.94 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 29 (114 mg, 0.47 mmol) and anhydrous K2C03 (260 mg, 1.88 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2-methoxyphenyl)-l,4-diazepan-l - yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one (compound 11) (40 mg, 17%). HPLC: 95%, RT 2.400 min. MS (ESI) m/z 422.2 [M + H]+. Synthesis of Compound 12
Scheme 22. S nthesis of intermediate 29
Figure imgf000064_0001
Intermediate 30 Intermediate 31
Scheme 23. Synthesis of compound 12
Figure imgf000064_0002
Intermediate 30:
Figure imgf000064_0003
[0154] A well-dried flask was first charged with l-bromo-2-ethoxybenzene (2.5 g, 12.4 mmol) and 1,4-diazepane (1.5 g, 15.0 mmol), which was evacuated and backfilled with N2 through a balloon under gentle warming (40°C). Toluene was charged and the mixture was bubbled with N2 for 10 min, then BINAP (234 mg, 0.38 mmol) and Pd2dba3 (1 19 mg, 0.13 mmol) was added to the mixture. After the addition of DBU (2.25 mL), the solution was warmed at 60- 70°C while a fine powder of ffluONa (2.1 g, 18.75 mmol) was added in one portion to start the amination. After the reaction mixture cooled to rt, (Boc)20 (6.9 g, 31.25 mmol) was dissolved in DCM and added dropwise to the reaction mixture, then stirred for 3 h at rt. The reaction mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 30:1) to give tert-bvXy\ 4-(2- ethoxyphenyl)-l,4-diazepane-l-carboxylate (intermediate 30) (800 mg, 20%). HPLC: 99%, RT 2.485 min. MS (ESI) m/z 321.0 [M + H]+.
Intermediate 31 :
Figure imgf000064_0004
[0155] Excess HC1 in EtOAc was added dropwise to a solution of intermediate 30 (800 mg, 2.5 mmol) in EtOAc and the reaction mixture was stirred at rt for 1.5 h. Filtration gave l-(2- ethoxyphenyl)-l,4-diazepane hydrochloride (intermediate 31) (382 mg, 59%) as a white solid.
Compound 12:
[0156] A mixture of intermediate 4 (150 mg, 0.5 mmol) and Nal (150 mg, 1.0 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 31 (129 mg, 0.5 mmol) and anhydrous K2CO3 (276 mg, 2.0 mmol) were then added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2-ethoxyphenyl)-l,4-diazepan-l- yl)butoxy)-3,4-dihydroquinolin-2(lH)-one (compound 12) (95 mg, 44%). ¾ MR (300 MHz, CDC13) δ 7.75 (bs, 1H), 7.04 (d, J= 8.1 Hz, 1H), 6.92-6.83 (m, 4H), 6.52 (dd, J = 8.4 Hz, 2.1 Hz, 1H), 6.30 (d, J = 2.7 Hz, 1H), 4.04 (q, J = 6.9 Hz, 2H), 3.96-3.94 (m, 2H), 3.38-3.29 (m, 4H), 3.02-2.93 (m, 4H), 2.89 (t, J= 6.9 Hz, 2H), 2.74-2.70 (m, 2H), 2.61 (t, J= 8.1 Hz, 2H), 2.14-2.10 (m, 2H), 1.83-1.79 (m, 4H), 1.45 (t, J= 6.9 Hz, 3H). HPLC: 99%, RT 2.438 min. MS (ESI) m/z 438.2 [M + H]+.
Synthesis of Compound 13
Scheme 24. Synthesis of compound 13
Figure imgf000065_0002
Compound 13:
Figure imgf000066_0001
[0157] A mixture of intermediate 12 (150 mg, 0.48 mmol) and Nal (150 mg, 1.0 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 31 (124 mg, 0.48 mmol) and anhydrous Κ2003 (265 mg, 1.92 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2-ethoxyphenyl)-l,4-diazepan-l- yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one (compound 13) (50 mg, 24%). XR NMR (300 MHz, CDCI3) δ 7.55 (bs, 1H), 7.35 (d, J= 8.1 Hz, 1H), 6.89-6.85 (m, 4H), 6.34 (d, J= 8.1 Hz, 1H), 4.24-4.20 (m, 2H), 4.04 (q, J= 6.9 Hz, 2H), 3.41-3.37 (m, 2H), 3.31 (t, J= 5.7 Hz, 2H), 3.15-2.95 (m, 4H), 2.86 (t, J= 6.9 Hz, 2H), 2.79-2.69 (m, 2H), 2.64 (t, J= 7.5 Hz, 2H), 2.20-2.12 (s, 2H), 1.82-1.76 (m, 4H), 1.45 (t, J = 6.9 Hz, 3H). HPLC: 99%, RT 2.393 min. MS (ESI) m/z 439.2 [M + H]+.
Synthesis of Compound 14
Scheme 25. Synthesis of compound 14
Figure imgf000066_0002
Compound 14:
Figure imgf000066_0003
[0158] A mixture of intermediate 15 (150 mg, 0.51 mmol) and Nal (150 mg, 1.0 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 31 (130 mg, 0.51 mmol) and anhydrous K2C03 (282 mg, 2.04 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2-ethoxyphenyl)-l,4-diazepan-l- yl)butoxy)quinolin-2(lH)-one (compound 14) (50 mg, 24%). lH NMR (300 MHz, CDC13) δ 10.89 (bs, 1H), 7.69 (d, J= 9.6 Hz, 1H), 7.44 (d, J= 9.0 Hz, 1H), 6.92-6.78 (m, 5H), 6.71 (d, J =2.7 Hz, 1H), 6.51 (d, J=9.6 Hz, 1H), 4.10-4.01 (m, 4H), 3.36-3.30 (m, 4H), 2.89-2.79 (m, 4H), 2.62-2.59 (m, 4H), 2.02-2.01 (m, 2H), 1.89-1.82 (m, 2H), 1.45 (t, J= 6.9 Hz, 3H). HPLC: 99%, RT 2.375 min. MS (ESI) m/z 436.2 [M + H]+.
Synthesis of Compound 15
Scheme 26. Synthesis of intermediate 32
Figure imgf000067_0001
Scheme 27. Synthesis of compound 15
Figure imgf000067_0002
Intermediate 32:
Figure imgf000067_0003
[0159] A mixture of intermediate 14 (193 mg, 1.2 mmol), 1,3-dibromopropane (0.37 mL, 3.6 mmol) and anhydrous K2C03 (166 mg, 1.2 mmol) was dissolved in EtOH and the solution was heated to reflux overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE EtOAc = 2:1) to give 7-(3-bromopropoxy)quinolin-2(lH)-one (intermediate 32) (185 mg, 55%) as a white solid. Compound 15:
Figure imgf000068_0001
[0160] A mixture of intermediate 32 (217 mg, 0.77 mmol) and Nal (231 mg, 1.54 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 2 (217 mg, 0.77 mmol) and anhydrous K2C03 (425 mg, 3.08 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 7-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)propoxy)quinolin-2(lH)-one (compound 15) (135 mg, 39%). 1H NMR (300 MHz, CDC13) δ 11.96 (bs., 1H), 7.72 (d, J = 9.3 Hz, 1H) 7.45 (d, J = 8.7 Hz, 1H), 7.09-7.07 (m, 2H), 7.01-6.99 (m, 1H), 6.83-6.81 (m, 2H), 6.54 (d, J= 9.0 Hz, 1H), 4.15 (t, J= 5.7 Hz, 2H), 3.31 (d, J= 5.1 Hz, 4H), 2.92-2.96 (m, 4H), 2.76 (t, J= 6.6 Hz, 2H), 2.03-1.01 (m, 4H). HPLC: 99%, RT 2.348 min. MS (ESI) m/z 446.1 [M + H]+. mp: 117-118°C.
Synthesis of Compound 16
Scheme 28. S nthesis of intermediate 34
Figure imgf000068_0002
Intermediate 33 Intermediate 34
Scheme 29. Synthesis of compound 16
Figure imgf000068_0003
Intermediate 33:
Figure imgf000068_0004
[0161] A well-dried flask was first charged with l-bromo-2-isopropoxybenzene (500 mg, 2.32 mmol) and 1,4-diazepane (348 mg, 3.48 mmol), which was evacuated and backfilled with N2 through a balloon under gentle warming (40°C). Toluene was charged and the mixture was bubbled with N2 for 10 min, then BINAP (44 mg, 0.07 mmol) and Pd2dba3 (14 mg, 0.02 mmol) were added to the mixture. After the addition of DBU (0.5 mL), the solution was warmed at 60- 70°C while a fine powder of iBuONa (334 mg, 3.48 mmol) was added in one portion to start the amination. After the reaction mixture cooled to rt, (Boc)20 (1.265 g, 5.8 mmol) was dissolved in DCM and added dropwise to the reaction mixture then stirred for 3 h at rt. The reaction mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 30:1) to give tert-butyl 4-(2-isopropoxyphenyl)- 1,4-diazepane-l-carboxylate (intermediate 33) (172 mg, 22%).
Intermediate 34:
Figure imgf000069_0001
[0162] Excess HC1 in EtOAc was added dropwise to a solution of intermediate 33 (172 mg, 0.51 mmol) in EtOAc and the reaction mixture was stirred at rt for 1.5 h. Filtration gave 1- (2-isopropoxyphenyl)- 1,4-diazepane hydrochloride (intermediate 34) (175 mg, 100%) as a white solid.
Compound 16:
Figure imgf000069_0002
[0163] A mixture of intermediate 4 (75 mg, 0.25 mmol) and Nal (75 mg, 0.5 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 34 (100 mg, 0.37 mmol) and anhydrous K2C03 (138 mg, 1.0 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2-isopropoxyphenyl)-l,4-diazepan-l- yl)butoxy)-3,4-dihydroquinolin-2(lH)-one (compound 16) (81 mg, 72%). 1H NMR (300 MHz, CDC13) δ 7.72 (bs, 1H), 7.03 (d, J=8.1 Hz, 1H), 6.89-6.85 (m, 4H), 6.51 (d, J=8.1 Hz, 1H), 6.30 (s, 1H), 4.62-4.57 (m, 1H), 4.02-3.92 (m, 2H), 3.42-3.36 (m, 2H ), 3.32-3.28 (m, 2H), 3.07-3.00 (m, 4H), 2.89 (t, J = 7.2 Hz, 2H), 2.80-2.74 (m, 2H), 2.61 (t, J= 6.9 Hz, 2H), 2.20-2.10 (m, 2H), 1.90-1.76 (m, 4H), 1.35 (d, J = 6.0 Hz, 6H). HPLC: 99%, RT 2.506 min. MS (ESI) m/z 452.4 [M + H]+.
Synthesis of Compound 17
Scheme 30. Synthesis of compound 17
Figure imgf000070_0001
Compound 17:
Figure imgf000070_0002
[0164] A mixture of intermediate 12 (114 mg, 0.36 mmol) and Nal (109 mg, 0.73 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 34 (98 mg, 0.36 mmol) and anhydrous K2C03 (200 mg, 1.45 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2-isopropoxyphenyl)-l,4-diazepan-l- yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one (compound 17) (52 mg, 32%). Ή NMR (300 MHz, CDCI3) δ 7.58 (bs., 1H), 7.35 (d, J=8.1 Hz, 1H), 6.89-6.86 (m, 4H,), 6.34 (d, J=8.1 Hz, 1H), 4.62-4.57 (m, 1H), 4.22 (m, 2H), 3.42-3.36 (m, 2H), 3.30 (t, J = 6.3 Hz, 2H), 3.08-3.01 (m, 4H), 2.89-2.77 (m, 4H), 2.64 (t, J= 7.2 Hz, 2H), 2.21-2.11 (m, 2H), 1.90-1.72 (m., 4H), 1.35 (d, J= 6.0 Hz, 6H). HPLC: 99%, RT 2.463 min. MS (ESI) m/z 453.3 [M + H]+. Synthesis of Compound 18
Scheme 31. Synthesis of compound 18
Figure imgf000071_0001
Compound 18:
Figure imgf000071_0002
[0165] A mixture of intermediate 15 (120 mg, 0.41 mmol) and Nal (123 mg, 0.82 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 34 (111 mg, 0.41 mmol) and anhydrous K2C03 (226 mg, 1.64 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2-isopropoxyphenyl)-l,4-diazepan-l- yl)butoxy)quinolin-2(lH)-one (compound 18) (115 mg, 62%). 1H NM (300 MHz, CDC13) δ 11.68 (bs., 1H), 7.71 (d, J= 9.3 Hz, 1H), 7.44 (d, J= 9.3 Hz, 1H), 6.89-6.79 (m, 6H), 6.53 (d, J= 9.6 Hz, 1H), 4.62-4.54 (m, 1H), 4.11-4.07 (m, 2H), 3.34-3.29 (m, 4H), 2.94-2.86 (m, 4H), 2.72- 2.62 (m, 2H), 2.12-1.96 (m, 2H), 1.87-1.76 (m, 4H), 1.35 (d, J= 6.0 Hz, 6H). HPLC: 99%, RT 2.434 min. MS (ESI) m/z 450.3 [M + H]+.
\ Synthesis of Compound 19
Scheme 32. Synthesis of intermediate 39
Figure imgf000072_0001
intermediate 35 intermediate 36
Figure imgf000072_0002
Scheme 33. Synthesis of compound 19
Figure imgf000072_0003
Example 19
Intermediate 35:
Figure imgf000072_0004
[0166] To a solution of 4-methoxy-2-nitroaniline (10 g, 59.4 mmol) in EtOH (200 mL) and water (80 mL) was added iron (12 g, 210 mmol) and ammonium chloride (9.63 g, 180 mmol). The mixture was then heated to reflux and stirred overnight. The mixture was filtered and the filtrate was evaporated to remove the EtOH and the residue was purified by flash chromatography on silica gel column (elution with DCM/MeOH = 80:1) to give 4- methoxybenzene-l,2-diamine (intermediate 35) (5.65 g, 69%) as a yellow solid.
Intermediate 36:
Figure imgf000072_0005
[0167] Intermediate 35 (5.6 g, 40.5 mmol) was dissolved in MeOH (10 mL). To this solution was added (Boc)20 (8.86 g, 40.6 mmol) followed by iodine (515 mg, 4.06 mmol), which was then stirred at rt for 1 h. The solvent was evaporated, and the residue was dissolved in EtOAc and washed with 5% aq Na2S203 and saturated aq NaHC03. The organic layer was dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 100:1) to give tert-butyl 2-amino-4- methoxyphenylcarbamate (intermediate 36) (7.4 g, 69%) as a brown solid, mp: 108-110°C.
Intermediate 37:
Figure imgf000073_0001
[0168] Intermediate 36 (7.3 g, 30.6 mmol) was dissolved in DMF (70 mL). To this solution was added anhydrous K2C03 (4.7 g, 33.7 mmol) and the mixture was heated to reflux (130°C) for 5 h. The solvent was removed under pressure, the residue was poured into ice water, and the aq phase was extracted with EtOAc. The combined organic layers were dried over anhydrous Na2S04, concentrated under vacuum, and the resulting yellow solid was recrystallized from EtOH to give 5-methoxy-lH-benzo[^imidazol-2(3H)-one (intermediate 37) (3.6 g, 72%) as a yellow solid, mp: 249-251°C.
Intermediate 38:
Figure imgf000073_0002
[0169] A mixture of intermediate 37 (3.6 g, 22 mmol) and 48% hydrogen iodide (30 mL) was heated to reflux (130°C) and stirred for 12 h. The reaction mixture was neutralized to pH = 8 with aq NaOH and then extracted with EtOAc. The combined organic layers were dried over anhydrous Na2S04, concentrated under vacuum and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 80: 1) to give 5-hydroxy-lH-benzo[cf]imidazol-2(3H)- one (intermediate 38) (16.9 g, 80%). HPLC: 99%, RT 1.377 min. MS (ESI) m/z 151.0 [M + H]+. mp: 186-187°C
Intermediate 39:
Figure imgf000073_0003
[0170] A solution of intermediate 38 (5 g, 33.3 mmol), 1,3-dibromopropane (4 mL, 33.3 mmol) and anhydrous K2C03 (4.6 g, 33.3 mmol) in EtOH (25 mL) was heated to reflux and stirred overnight. The solvent was removed under vacuum. The residue was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with water, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 100: 1) to give 5-(3-bromopropoxy)-lH-benzo[d]imidazol- 2(3H)-one (intermediate 39) (240 mg, 12%) as a yellow solid. Compound 19:
Figure imgf000074_0001
[0171] Intermediate 39 (80 mg, 0.29 mmol) was dissolved in CH3CN (9 mL), then Nal (89 mg, 0.59 mmol) was added and the mixture was heated to reflux for 30 min, then cooled to rt. Intermediate 2 (43 mg, 0.15 mmol) and anhydrous K2C03 (163 mg, 1.19 mmol) were added. The resulting mixture was heated to reflux and stirred for 6 h. The reaction solution was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 5:1) to give 5-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)propoxy)-lH-benzo[iflimidazol-2(3H)-one (compound 19) (41mg, 32%). Ή NMR (300 MHz, CDC13): δ 9.37 (bs, 1H), 9.05 (bs., 1H), 7.09 - 7.07 (m, 2H), 7.00 - 6.97 (m, 1H), 6.91 - 6.88 (m, 1H), 6.66 - 6.60 (m, 2H), 4.10 - 3.90 (m, 2H), 3.31 - 3.29 (m, 4H), 2.95 - 2.80 (m, 4H), 2.76 - 2.71 (m, 2H), 2.04 - 1.92 (m, 4H). HPLC: 99%, RT 2.241 min. MS (ESI) m/z 436.9 [M + H]+. mp: 148-150°C.
Synthesis of Compound 20
Scheme 34. Synthesis of intermediate 41
Figure imgf000074_0002
Intermediate 40 Intermediate 41
Scheme 35. Synthesis of intermediate 44
Figure imgf000074_0003
Intermediate 43
Figure imgf000074_0004
Intermediate 40:
Figure imgf000075_0001
[0172] A well-dried flask was first charged with l-bromo-2,3-dichlorobenzene (10 g, 44 mmol) and piperazine (4.55 g, 53 mmol), which was evacuated and backfilled with N2 through a balloon under gentle warming (40°C). Toluene was charged and the mixture was bubbled with N2 for 10 min, then BINAP (822 mg, 1.32 mmol) and Pd2dba3 (403 mg, 0.44 mmol) were added to the mixture. After the addition of DBU (8.0 mL), the solution was warmed at 60-70°C while a fine powder of ffluONa (7.39 g, 66 mmol) was added in one portion to start the amination. After the reaction mixture cooled to rt, (Boc)20 (24 g, 110 mmol) was dissolved in DCM and added dropwise to the reaction mixture, then stirred at rt for 3 h. The reaction mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 10:1) to give tert-butyl 4-(2,3-dichlorophenyl)piperazine-l- carboxylate (intermediate 40) (11.1 g, 76%) as a white solid.
Intermediate 41:
Figure imgf000075_0002
[0173] Excess HC1 in EtOAc was added dropwise to a solution of intermediate 40 (11.1 g, 33.5 mmol) in EtOAc and the reaction mixture was stirred at rt for 1.5 h. Filtration gave 1- (2,3-dichlorophenyl)piperazine hydrochloride salt (intermediate 41) (7.5 g, 78%) as a white solid. HPLC: 99%, RT 2.052 min. MS (ESI) m/z 231.0[M + H]+.
Intermediate 42:
Figure imgf000075_0003
[0174] A mixture of 2-fluoro-4-methoxybenzaldehyde (1 g, 6.5 mmol) and hydrazine (7 mL) was heated to reflux for 4 h. The mixture was extracted with CH2(¾. The combined organic layers were washed with water, brine, concentrated in vacuo, and purified by column chromatography (elution with PE/EtOAc=5: l) to afford 6-methoxy-lH-indazole (intermediate 42) (568 mg, 59%) as a yellow solid. HPLC: 99%, RT 2.159 min. MS (ESI) m/z 149.1[M + H]+. Intermediate 43:
Figure imgf000075_0004
[0175] A mixture of 6-methoxy-lH-indazole (560 mg, 3.8 mmol) and HI (2.4 mL 48%) was heated to reflux for 4 h. The mixture was extracted with CH2C12. The combined organic layers were washed with water, brine, concentrated in vacuo, and purified by column chromatography (CH2Cl2 MeOH = 60/1) to give lH-indazol-6-ol (intermediate 43) (436 mg, 86%).
Intermediate 44:
Figure imgf000076_0001
[0176] A mixture of lH-indazol-6-ol (600 mg, 4.5 mmol), 1,3-dibromopropane (1.4 mL, 13.4 mmol) and anhydrous K2C03 (618 mg, 4.5 mmol) in EtOH (20 mL) was heated to reflux and stirred overnight. Filtration gave a yellow solid which was purified by column chromatography (elution with PE/EtOAc = 3:1) to afford 6-(3-bromopropoxy)-lH-indazole (intermediate 44) (84 mg, 7%) as a yellow oil.
Compound 20:
Figure imgf000076_0002
[0177] A mixture of intermediate 44 (84 mg, 0.31 mmol) and Nal (94 mg, 0.63 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (125 mg, 0.47 mmol) and anhydrous K2C03 (172 mg, 1.25 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 40:1) to give 6-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)-lH-indazole (compound 20)
(90 mg, 71%). 'H NMR(300 MHZ, CDC13): δ 7.97 (s, 1H), 7.61 (d, J - 8.7 Hz, 1H), 7.16 - 7.14 (m, 2H), 6.98 - 6.95 (m, 1H), 6.87 - 6.82 (m, 2H), 4.10 (t, J = 6.6Hz, 2H), 3.10 (m, 4H), 2.71 - 2.63 (m, 6H), 2.10 - 2.04 (m, 2H). HPLC: 99%, RT 2.462 min. MS (ESI) m/z 405.2 [M + H]+. mp: 159-161°C. Synthesis of Compound 21
Scheme 37. Synthesis of intermediate 47
Figure imgf000077_0001
H2,RhCI(PPh3)3
96%
toluene 70°C, 4d
Figure imgf000077_0002
intermediate 47 Intermediate 46 38. Synthesis of compound 21
Figure imgf000077_0003
Intermediate 45:
Figure imgf000077_0004
[0178] To a -30°C solution of l-bromo-2,3-dichlorobenzene (10 g, 44 mmol) in THF (120 mL) was added z-PrMgCl (2.0 M in THF, 35 mL, 70 mmol) at a rate such that the temperature < -20°C. Meanwhile, to a -10°C solution of Cul (420 mg, 2.2 mmol) in THF (120 mL) was added pyridine (7.1 mL, 88 mmol) and then benzyl carbonochloridate (9.7 mL, 68 mmol) such that the temperature < 0°C. To this heterogeneous mixture was added the initially formed Grignard at a rate such that the temperature < 0°C. The resulting solution was stirred at 0°C for 30 min and then allowed to warm to rt. The reaction was then quenched with 10% aq NH4C1. EtOAc was added and the blue aq layer was removed. The organic layer was washed with 10% aq H4CI, 1 N HC1, and a 20% aq NaCl solution. The organic layer was then concentrated and the residue was dissolved and crystallized from MeOH. The slurry was filtered and the filtercake washed with MeOH to give benzyl 4-(2,3-dichlorophenyl)pyridine-l(4H)- carboxylate (intermediate 45) (12 g, 76%) as an off-white solid. HPLC: 99%, RT 4.118 min. MS (ESI) m/z 382.0 [M + Na]+. mp: 69-70°C.
Intermediate 46:
Figure imgf000078_0001
[0179] To a solution of intermediate 45 (7.0 g, 19.5 mmol) in toluene (150 mL) was added R Cl(PPh3)3 (2.1 g, 2.0 mmol) as a slurry in toluene (50 mL). The reaction was subjected to an atmosphere of H2 at 40 psi and heated to 70°C. After 6 h, the reaction was filtered through silica gel and the silica gel was washed with 1 :9 EtO Ac/toluene. The filtrate was dissolved in toluene, concentrated in vacuo, and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 50:1) to give benzyl 4-(2,3-dichlorophenyl)piperidine-l-carboxylate (intermediate 46) (6.8 g, 96%) as an oil. HPLC: 99%, RT 30843 min. MS (ESI) m/z 386.1 [M + Na]+.
Intermediate 47:
Figure imgf000078_0002
[0180] To 6 N HC1 (30 mL) was added a solution of intermediate 46 (5.2 g, 14 mmol) in THF (10 mL). The mixture was heated to reflux for 3 h and then concentrated in vacuo. The residue was washed with Et20 to give 4-(2,3-dichlorophenyl)piperidine (intermediate 47) (3.5 g, 92%) as a white solid. HPLC: 99%, RT 2.149 min. MS (ESI) m/z 230.1 [M + H]+
Compound 21:
Figure imgf000078_0003
[0181] A mixture of intermediate 26 (114 mg, 0.4 mmol) and Nal (120 mg, 0.8 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 47 (160 mg, 0.6 mmol) and anhydrous K2C03 (221 mg, 1.6 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 5-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butoxy)benzo[d]thiazole (compound 21) (144 mg, 66%). ¾ MR (300 MHz, CDC13): δ 8.97 (s, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.61 (d, J = 2.1 Hz, 1H), 7.33-7.29 (m, 1H), 7.26-7.16 (m, 1H), 7.09 (dd, J= 2.1 Hz, 8.7 Hz, 3H), 4.10 (t, J = 6.6 Hz, 2H), 3.13-3.09 (m, 2H), 2.50 (t, J=7.5 Hz, 2H), 2.14-2.10 (m, 2H), 1.91-1.74 (m, 10H). HPLC: 99%, RT 2.689 min. MS (ESI) m/z 435.3 [M + H]+. mp: 79-81°C.
Synthesis of Compound 22
Scheme 39. Synthesis of compound 22
Figure imgf000079_0001
[0182] A mixture of intermediate 26 (218 mg, 0.8 mmol) and Nal (240 mg, 1.6 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 47 (320 mg, 1.2 mmol) and anhydrous K2C03 (442 mg, 3.8 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 5-(3-(4-(2,3-dichlorophenyl)piperidin-l-yl)propoxy)benzo[d]thiazole (compound 22) (21 1 mg, 56%). Ή NMR (300 MHz, CDC13): δ 8.98 (s, 1H), 7.80 (d, J = 8.7 Hz, 1H), 7.62 (d, J = 3.0 Hz, 1H), 7.34-7.31 (m, 1H), 7.24-7.18 (m, 3H), 7.15-7.09 (m, 1H), 4.15 (t, J = 5.7Hz, 2H), 3.18-3.12 (m, 3H), 2.70-2.66 (m, 2H), 2.27-2.19 (m, 2H), 2.17-2.10 (m, 2H), 1.92-1.81 (m, 4H). HPLC: 99%, RT 2.622 min. MS (ESI) m/z 421.1 [M + H]+. mp: 114-1 16°C. Synthesis of Compound 23
Scheme 40. Synthesis of compound 23
Figure imgf000080_0001
Intermediate 27 xamp e
Compound 23:
Figure imgf000080_0002
[0183] A mixture of intermediate 27 (218 mg, 0.8 mmol) and Nal (240 mg, 1.6 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (320 mg, 1.2 mmol) and anhydrous K2C03 (442 mg, 3.2 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 5-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)benzo[d]thiazole (compound 23) (209 mg, 62%). 1H NMR(300 MHz, CDC13): δ 8.97 (s, 1H), 7.80 (d, J= 8.7 Hz, 1H), 7.63 (d, J = 2.1 Hz, 1H), 7.16-7.09 (m, 3H), 6.94 (dd, J = 3.6 Hz, 6.0 Hz, 1H), 4.15(t, J = 5.7Hz, 2H), 3.09 (m, 4H), 2.68-2.63 (m, 6H), 2.10-2.05 (m, 2H). HPLC: 99%, RT 2.607 min. MS (ESI) m/z 422.0 [M + H]+. mp: 127-129°C.
Synthesis of Compound 24
Scheme 41. S nthesis of intermediate 48
Figure imgf000080_0003
Intermediate 38 reflux Scheme 42. Synthesis of compound 24
Figure imgf000081_0001
Intermediate 48:
Figure imgf000081_0002
[0184] A mixture of intermediate 38 (253 mg, 1.68 mmol), 1 ,4-dibromobutane (0.6 mL, 5.04 mmol) and anhydrous K2C03 (232 mg, 1.68 mmol) was dissolved in EtOH and the solution was heated to reflux overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 4:1) to give 7-(4-bromobutoxy)quinolin-2(lH)-one (intermediate 48) (120 mg, 25%) as a yellow solid.
Compound 24:
Figure imgf000081_0003
[0185] A mixture of intermediate 48 (200 mg, 0.7 mmol) and Nal (210 mg, 1.4 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 47 (167 mg, 0.63 mmol) and anhydrous K2C03 (386 mg, 2.8 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous a2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 5:1) to give 5-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butoxy)-lH-benzo[d]imidazol-2(3H)-one
(compound 24) (61 mg, 20%). !H NMR(300 MHz, CDC13): δ 9.59 (brs, 1H), 9.29 (brs, 1H), 7.33-7.30 (m, 2H), 7.20-7.13 (m, 1H), 6.92 (d, J = 8.7 Hz, 1H), 6.65-6.59 (m, 2H), 3.94-3.91 (m, 2H), 3.15-3.04 (m, 3H), 2.51-2.49 (m, 2H), 2.15(t, J = 11.1Hz, 2H), 1.90-1.74 (m, 8H). HPLC: 99%, RT 2.332 min. MS (ESI) m/z 434.0 [M + H]+. mp: 180-182°C. Synthesis of Compound 25
Scheme 43. Synthesis of compound 25
Figure imgf000082_0001
Compound 25:
Figure imgf000082_0002
[0186] A mixture of intermediate 21 (220 mg, 0.74 mmol) and Nal (222 mg, 1.48 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (278 mg, 1.04 mmol) and anhydrous K2C03 (1409 mg, 2.96 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)-3,4-dihydroisoquinolin-l(2H)- one (compound 25) (215 mg, 65%). Ή NMR (300 MHz, CDC13): δ 7.58 (d, J = 2.7 Hz, 1H), 7.14-7.11 (m, 3H), 7.01-6.94 (m, 2H), 6.18 (brs, 1H), 4.05 (t, J= 6.3 Hz, 2H), 3.56-3.51 (m, 2H), 3.07 (m, 4H), 2.90 (t, J = 8.1 Hz, 2H), 2.66 (m, 4H), 2.49-2.46 (m, 2H), 1.86-1.68 (m, 4H). HPLC: 99%, RT 2.374 min. MS (ESI) m/z 448.3 [M + H]+.
Synthesis of Compound 26
Scheme 44. Synthesis of intermediate 49
Figure imgf000082_0003
Intermediate 49 Scheme 45. Synthesis of compound 26
Figure imgf000083_0001
Intermediate 49 CH3CN Example 26
Intermediate 49:
Figure imgf000083_0002
[0187] A mixture of 7-hydroxy-3,4-dihydroquinolin-2(lH)-one (196 mg, 1.2 mmol), 1,4- bis(bromomethyl)cyclohexane (972 mg, 3.6 mmol) and anhydrous K2CO3 (166 mg, 1.2 mmol) was dissolved in EtOH and the solution was heated to reflux overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column to give 7-((4-(bromomethyl)cyclohexyl)methoxy)-3,4- dihydroquinolin-2(lH)-one (intermediate 49) (130 mg, 30%) as a white solid.
Compound 26:
Figure imgf000083_0003
[0188] A mixture of intermediate 49 (70 mg, 0.2 mmol) and Nal (60 mg, 0.4 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (81 mg, 0.3 mmol) and anhydrous K2C03 (110 mg, 0.8 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-((4-((4-(2,3-dichlorophenyl)piperazin-l-yl)methyl)cyclohexyl)methoxy)-3,4-dihydroquinolin-
2(lH)-one (compound 26) (63 mg, 63%). 1H NMR (300 MHz, CDC13): δ 8.06 (s, 1H), 7.13-7.15 (m, 2H), 7.04 (d, J= 8.4 Hz, 1H), 6.94-6.97 (m, 1H), 6.52 (dd, J= 2.4 Hz, 8.4 Hz, 1H), 6.32 (d, J = 2.1 Hz, 1H), 3.71-3.74 (m, 2H), 3.05-3.07 (m, 4H), 2.87-2.92 (m, 2H), 2.59-2.64 (m, 6H), 2.24 (d, J= 6.9 Hz, 2H), 3.83 (d, J= 9.9 Hz, 2H), 1.53-1.69 (m, 6H), 0.94-1.10 (m, 4H). HPLC: 99%, RT 2.652 min. MS (ESI) m/z 502.2 [M + H]+. Synthesis of Compound 27
Scheme 46. Synthesis of intermediate 50
Figure imgf000084_0001
Intermediate 50
Scheme 47. Synthesis of compound 27
Figure imgf000084_0002
Example 27
Intermediate 50:
Figure imgf000084_0003
[0189] A mixture of 7-hydroxy-3,4-dihydroquinolin-2(lH)-one (930 mg, 6 mmol), 1,4- dichlorobut-2-yne (326 mg, 2 mmol) and anhydrous K2C03 (276 mg, 2 mmol) was dissolved in EtOH and the solution was heated to reflux overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column to give 7-(4-chlorobut-2-ynyloxy)-3,4-dihydroquinolin-2(lH)-one (intermediate 50) (330 mg, 66%) as a yellow solid.
Compound 27:
Figure imgf000084_0004
[0190] A mixture of intermediate 50 (200 mg, 0.8 mmol) and Nal (240 mg, 1.6 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (320 mg, 1.2 mmol) and anhydrous K2C03 (442 mg, 3.2 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give 7-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)but-2-ynyloxy)-3,4-dihydroquinolin- 2(lH)-one (compound 27) (280 mg, 79%). Ή NMR (300 MHz, CDC13): δ 7.85 (s, 1H), 7.20- 7.13 (m, 2H), 7.08 (d, J= 8.4 Hz, 1H), 6.97 (dd, J = 3.0Hz, 6.6Hz, 1H), 4.73 (s, 2H), 3.42 (s, 2H), 3.10 (s, 4H), 2.93-2.88 (m, 2H), 2.75 (s, 4H), 2.64-2.59 (m, 2H), 2.06 (s, lH), 1.70 (s, 2H), 1.29- 1.24 (m, 1H). HPLC: 99%, RT 2.403 min. MS (ESI) m/z 444.1 [M + H]+.
Synthesis of Compound 28
esis of intermediate 51
Figure imgf000085_0001
Scheme 49. Synthesis of compound 28
Figure imgf000085_0002
Intermediate 51:
Figure imgf000085_0003
[0191] A mixture of lH-indazol-6-ol (intermediate 43) (320 g, 2.39 mmol), 1 ,4- dibromobutane (0.9 mL, 7.2 mmol), anhydrous K2C03 (55 mg, 2.39 mmol) and EtOH (5 mL) was heated to reflux and stirred overnight. A yellow solid was filtered and purified by column chromatography (elution with PE/EtOAc = 3 :1) to afford 5-(3-bromopropoxy)benzo[d]thiazole (intermediate 51) (70 mg, 17%) as a yellow oil.
Compound 28:
Figure imgf000085_0004
[0192] A mixture of intermediate 51 (65 mg, 0.24 mmol) and Nal (72 mg, 0.48 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (97 mg, 0.36 mmol) and anhydrous K2C03 (132 mg, 0.96 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 6-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)-lH-indazole (compound 28) (35 mg, 35%). Ή MR (300 MHz, CDC13): δ 7.97 (s, 1H), 7.61 (d, J = 8.7 Hz 1H), 7.17-7.15 (m, 2H), 6.96(dd, J = 3.0 Hz, 6.6 Hz, 1H), 6.86-6.81 (m, 2H), 4.05 (t, J = 5.4 Hz, 2H), 3.16 (s, 4H), 2.79 (s, 4H), 2.64-2.60 (m, 2H), 1.89-1.83 (m, 4H). HPLC: 99%, RT 2.491 min. MS (ESI) m/z 419.2 [M + H]+. mp: 101-103°C.
Synthesis of Compound 29
Scheme 50. Synthesis of intermediate 52
Figure imgf000086_0001
Intermediate 52
Scheme 51. Synthesis of compound 29
Figure imgf000086_0002
Intermediate 52:
Figure imgf000086_0003
[0193] A mixture of 7-hydroxy-3,4-dihydroquinolin-2(lH)-one (196 mg, 1.2 mmol), (£)- 1 ,4-dichlorobut-2-ene (450 mg, 3.6 mmol) and anhydrous K2C03 (166 mg, 1.2 mmol) was dissolved in EtOH and the solution was heated to reflux and stirred overnight. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column to give (£)-7-(4-chlorobut-2-enyloxy)-3,4- dihydroquinolin-2(lH)-one (intermediate 52) (102 mg, 34%) as a white solid.
Compound 29:
Figure imgf000086_0004
[0194] A mixture of intermediate 52 (75 mg, 0.25 mmol) and Nal (75 mg, 0.5 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (102 mg, 0.38 mmol) and anhydrous K2C03 (138 mg, 1 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2SC>4, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give (E)-7-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)but-2-enyloxy)-3,4-dihydroquinolin- 2(lH)-one (compound 29) (79 mg, 71%). ]H MR (300 MHz, CDC13): δ 7.77 (s, 1H), 7.16-7.14 (m, 2H), 7.05 (d, J= 8.1 Hz, 1H), 6.98-6.94 (m, 1H), 6.53 (d, J- 8.1 Hz, 1H), 6.33 (s, 1H), 5.94- 5.89 (m, 2H), 4.51 (d, J= 4.2 Hz, 2H), 3.19-3.08 (m, 6H), 2.90 (t, J= 6.9 Hz, 2H), 2.67-2.59 (m, 6H). HPLC: 99%, RT 2.468 min. MS (ESI) m/z 446.1 [M + H]+. mp: 151-152°C.
Synthesis of Compound 30
Scheme 52. Synthesis of intermediate 53
Figure imgf000087_0001
Intermediate 41 Intermediate 53
Scheme 53. Synthesis of compound 30
Figure imgf000087_0002
Intermediate 53:
Figure imgf000087_0003
[0195] A mixture of l-(2,3-dichlorophenyl)piperazine hydrochloride (intermediate 41) (294 mg, 1.1 mmol), l-bromo-3-(bromomethyl)benzene (250 mg, 1 mmol) and anhydrous triethylamine (253 mg, 2.5 mmol) was dissolved in CH3CN and the solution was heated to reflux for 4 h. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 8:1) to give l-(3-bromobenzyl)-4-(2,3-dichlorophenyl)piperazine (intermediate 53) (220 mg, 74%) as a white solid. Compound 30:
Figure imgf000088_0001
[0196] To a solution of 7-hydroxy-3,4-dihydroquinolin-2(lH)-one (196 mg, 1.2 mmol in NMP was added Cs2C03 (391 mg, 1.2 mmol). The slurry was degassed by evacuating and filling the reaction flask with N2 three times. Intermediate 53 (240 mg, 0.6 mmol) and TMHD (11 mg, 0.06 mmol) were added followed by the addition of CuCl (60 mg, 0.6mmol). The reaction mixture was degassed by evacuating and filling the reaction flask with N2 three times, and then warmed to 120°C under N2 for 7.5 h. The reaction mixture was cooled to rt and diluted with Et20. The slurry was filtered and the filtercake was washed with Et20. Combined filtrates were washed with 2 N HC1, 0.6 N HC1, 2 M NaOH, and 10% aq NaCl. The resulting organic layer was dried, concentrated, and purified by flash chromatography on a silica gel column (elution with PE/EtOAc = 1 :1) to give 7-(3-((4-(2,3-dichlorophenyl)piperazin-l-yl)methyl)phenoxy)-3,4- dihydro quinolin-2(lH)-one (compound 30) (110 mg, 38%). 'H NMR (300 MHz, CDC13): δ 7.62 (s, 1H), 7.31 (d, J = 7.2 Hz, 2H), 7.15-7.05 (m, 4H), 6.96-6.89 (m, 2H), 6.62 (dd, J = 2.1 Hz, 8.1Hz, 1H), 6.41 (d, J= 2.1Hz, 1H), 3.57 (s, 2H), 3.06 (m, 4H), 2.94 (t, J= 7.5Hz, 2H), 2.66-2.61 (m, 5H). HPLC: 99%, RT 2.637 min. MS (ESI) m/z 482.1 [M + H]+. mp: 182-183°C.
Synthesis of Compound 31
Scheme 54. Synthesis of intermediate 54
Figure imgf000088_0002
E l 1 Intermediate 54:
r
Figure imgf000089_0001
[0197] A mixture of intermediate 41 (294 mg, 1.1 mmol), l-bromo-4-(bromomethyl) benzene (250 mg, 1.0 mmol) and anhydrous triethylamine (253 mg, 2.5 mmol) was dissolved in CH3CN and the solution was heated to reflux for 4 h. The solution was diluted with water and extracted with EtOAc. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 8:1) to give l-(4-bromobenzyl)-4-(2,3- dichlorophenyl)piperazine (intermediate 54) (220 mg, 74%) as a white solid.
Compound 31:
Figure imgf000089_0002
[0198] To a solution of 7-hydroxy-3,4-dihydroquinolin-2(lH)-one (228 mg, 1.4 mmol) in NMP was added Cs2C03 (456 mg, 1.4 mmol). The slurry was degassed by evacuating and filling the reaction flask with N2 three times. Intermediate 54 (280 mg, 0.7 mmol) and TMHD (13 mg, 0.07 mmol) were added followed by the addition of CuCl (70 mg, 0.7 mmol). The reaction mixture was degassed by evacuating and filling the reaction flask with N2 three times, and then warmed to 120°C under N2 for 7.5 h. The reaction mixture was cooled to rt and diluted with Et20. The slurry was filtered and the filtercake was washed with Et20. Combined filtrates were washed with 2 N HC1, 0.6 N HC1, 2 M NaOH, and 10% aq NaCl. The resulting organic layer was dried, concentrated, and purified by flash chromatography on a silica gel column (elution with PE EtOAc = 1:1) to give 7-(4-((4-(2,3-dichlorophenyl)piperazin-l-yl)methyl)phenoxy)-3,4- dihydro quinolin-2(lH)-one (compound 31) (120 mg, 35%). !H NMR (300 MHz, CDC13): δ 7.67 (s, 1H), 7.32 (d, J = 8.7 Hz, 2H), 7.16-7.09 (m, 3H), 6.98-6.94 (m, 3H), 6.64-6.61 (m, 1H), 6.41 (d, J = 3.0Hz, 1H), 3.56 (s, 2H), 3.07 (m, 4H), 2.94 (t, J = 6.6 Hz, 2H), 2.66-2.61 (m, 6H). HPLC: 99%, RT 2.626 min. MS (ESI) m/z 482.1 [M + H]+. mp: 185-186°C. Synthesis of Compound 32
Scheme 56. Synthesis of compound 32
Figure imgf000090_0001
[0199] A mixture of intermediate 20 (110 mg, 0.39 mmol) and Nal (117 mg, 0.78 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (157 mg, 0.59 mmol) and anhydrous K2C03 (216 mg, 1.56 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 6 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)-3,4-dihydroisoquinolin-l(2H)- one (compound 32) (88 mg, 52%). ¾ NMR (300 MHz, CDC13): δ 7.61 (s, 1H), 7.14-7.11 (m, 3H), 7.03-6.98(m, 2H), 5.93 (brs, 1H), 4.11 (m, 2H), 3.55 (m, 2H), 3.09-2.94 (m, 6H), 2.65-2.61 (m, 6H), 2.02 (s, 2H). HPLC: 99%, RT 2.612 min. MS (ESI) m/z 434.1 [M + H]+. mp: 162- 164°C.
Synthesis of Compound 33
Scheme 57. Synthesis of compound 33
Figure imgf000090_0002
Compound 33:
Figure imgf000091_0001
[0200] A mixture of intermediate 21 (103 mg, 0.35 mmol) and Nal (105 mg, 0.7 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 47 (142 mg, 0.53 mmol) and anhydrous K2C03 (194 mg, 1.4 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30: 1) to give 7-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butoxy)-3,4-dihydroisoquinolin-l(2H)- one (compound 33) (108 mg, 69%). *H NMR (300 MHz, CDC13): δ 7.58 (d, J = 2.1Hz, 1H), 7.23-7.11 (m, 5H), 7.02-6.99 (m, 1H), 6.05 (s, 1H), 4.05 (t, J = 6.0 Hz, 2H), 3.56-3.53 (m, 2H), 3.15-3.06 (m, 3H), 2.93 (t, J = 6.6Hz, 2H), 2.51 (m, 2H), 2.18 (m, 2H), 1.82 (m, 8H). HPLC: 99%, RT 2.746 min. MS (ESI) m/z 447.2 [M + H]+. mp: 113-114°C.
Synthesis of Compound 34
Scheme 58. Synthesis of compound 34
Figure imgf000091_0002
in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (188 mg, 0.71 mmol) and anhydrous K2C03 (259 mg, 1.88 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 15:1) to give 5-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)-lH-benzo[d]imidazol-2(3H)- one (compound 34) (54 mg, 27%). 'H MR (300 MHz, DMSO): δ 10.60 (s, 1H), 10.36 (s, 1H), 7.32 (d, J = 5.1 Hz, 2H), 7.16-7.18 (m, 1H), 6.80 (d, J = 9.6 Hz, 1H), 6.51-6.53 (d, J = 6.6 Hz, 2H), 3.99-3.95 (m, 2H), 2.81-3.26 (s, 5H), 2.80-2.60 (m, 5H), 1.80-2.01 (m, 2H). HPLC: 99%, RT 2.228 min. MS (ESI) m/z 421.1 [M + H]+.
Synthesis of Compound 35
Scheme 59. Synthesis of compound 35
Figure imgf000092_0001
Compound 35:
Figure imgf000092_0002
[0202] A mixture of intermediate 48 (110 mg, 0.39 mmol) and al (117 mg, 0.78 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (156 mg, 0.59 mmol) and anhydrous K2C03 (215 mg, 1.56 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred overnight. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 15:1) to give 5-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)-lH-benzo[d]imidazol-2(3H)- one (compound 35) (54 mg, 32%). *H NMR (300 MHz, CDC13): δ 9.53 (s, 1H), 9.24 (s, 1H), 7.17-7.13 (m, 2H), 6.97-6.91 (m, 2H), 6.66-6.60 (m, 2H), 3.95 (t, J= 5.7 Hz, 2H), 3.20-3.01 (m, 4H), 2.80-2.61 (m, 4H), 2.52 (t, J= 7.5 Hz, 2H), 1.82-1.73(m, 4H). HPLC: 99%, RT 2.325 min. MS (ESI) m/z 435.1 [M + H]+. Synthesis of Compound 36
Figure imgf000093_0001
Compound 36:
Figure imgf000093_0002
[0203] A mixture of intermediate 26 (108 mg, 0.38 mmol) and Nal (114 mg, 0.76 mmol) in C¾CN was heated to reflux for 30 min and then cooled to rt. Intermediate 41 (152 mg, 0.57 mmol) and anhydrous K2C03 (210 mg, 1.52 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 5-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)benzo[d]thiazole (compound 36) (98 mg, 60%). ¾ NM (300 MHz, CDC13): δ 8.97 (s, 1H), 7.79 (d, J = 8.7 Hz, 1H), 7.61 (s, 1H), 7.16-7.08 (m, 3H), 6.97-6.94 (m, 1H), 4.11 (t, J= 5.7 Hz, 2H), 3.09 (br, 4H), 2.68 (brs, 4H), 2.55-2.50 (m, 2H), 1.92-1.76 ( m, 5H). HPLC: 99%, RT 2.651 min. MS (ESI) m/z 436.3 [M+H]+. mp: 93-94.5°C.
Synthesis of Compound 37
Scheme 61. Synthesis of intermediate 55
Figure imgf000093_0003
Intermediate 55
Scheme 62. S nthesis of com ound 37
Figure imgf000093_0004
xamp e Intermediate 55:
Figure imgf000094_0001
[0204] A well-dried flask was first charged with l-bromo-2-methoxybenzene (4.0 g, 21.3 mmol) and piperazine (2.2 g, 25.6 mmol), which was evacuated and backfilled with N2 through a balloon under gentle warming (40°C). Toluene was charged and the mixture was bubbled with N2 for 10 min, then BINAP (398 mg, 0.64 mmol) and Pd2dba3 (195 mg, 0.21 mmol) was added to the mixture. After the addition of DBU (3.8 mL), the solution was warmed at 60-70°C while a fine powder of tBuONa (3.5 g, 31.9 mmol) was added in one portion to start the amination. After the reaction mixture cooled to rt, (Boc)20 (11.6 g, 53.2 mmol) was dissolved in DCM and added dropwise to the reaction mixture, then stirred at rt for 3 h. The reaction mixture was diluted with water and extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with PE/EtOAc = 10:1) to give tert-butyl 4-(2-methoxyphenyl)piperazine-l- carboxylate as a white solid. Excess HC1 in EtOAc was added dropwise to a solution of tert- butyl 4-(2-methoxyphenyl)piperazine-l-carboxylate in EtOAc and the reaction mixture was stirred at rt for 1.5 h. Filtration gave l-(2-methoxyphenyl)piperazine hydrochloride salt (intermediate 55) (2.6 g, 42%) as a white solid. HPLC: 99%, RT 1.586 min. MS (ESI) m/z 193.1 [M + H]+.
Compound 37:
Figure imgf000094_0002
[0205] A mixture of intermediate 12 (314 mg, 0.48 mmol) and Nal (150 mg, 0.96 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 55 (228 mg, 0.48 mmol) and anhydrous K2C03 (138 mg, 1.92 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2-methoxyphenyl)piperazin-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one (compound 37) (98 mg, 60%). ¾ NMR (300 MHz, CDC13): δ 7.55 (s, 1H), 7.35 (d, J= 8.1 Hz, 1H), 7.02-6.85 (m, 3H), 6.37-6.34(m, 1H), 4.29-4.20 (m, 2H), 3.86 (s, 3H), 3.11 (brs, 4H), 2.86 (t, J = 6.9 Hz, 2H), 2.67-2.61(m, 5H), 2.48 (t, J = 7.2 Hz, 1H), 1.82-1.62(m, 6H). HPLC: 99%, RT 2.109 min. MS (ESI) m/z 411.2 [M + H]+.
Synthesis of Compound 38
Scheme 63. S nthesis of intermediate 59
Figure imgf000095_0001
Intermediate 57 Intermediate 58 Intermediate 59
Scheme 64. Synthesis of compound 38
Figure imgf000095_0002
[0206] To a cooled (0°C) solution of 2-amino-4-chlorophenol (30 g, 209 mmol) in DCM (200 mL) and pyridine (17 mL) was added a solution of chloroacetyl chloride in DCM (23.6 g, 230 mmol in 75 mL). The mixture was allowed to reach rt slowly and was subsequently stirred for 18 h. The precipitate was filtered, washed with DCM, and dried overnight at 50°C to give crude 2-chloro-N-(5-chloro-2-hydroxyphenyl)acetamide (intermediate 56) (26 g, 55%), as a brown solid which was used in the next step without any further purification.
Intermediate 57:
Figure imgf000095_0003
[0207] Intermediate 56 (26 g, 118 mmol) was suspended in water at 0°C. To suspension was added HN03 (1.5 equiv, 15 mL, 174 mmol) over a period of 0.75 h. resulting red solution was allowed to reach rt after 1 h, then water was added to give a dark brown solid which was isolated by filtration. The resulting filtrate was washed with cold water (3x100 mL) and dried to give a yellow solid. Ring closure to intermediate 57 was established by heating the yellow solid with K2C03 in toluene/water (50/1, v/v) for 2 h. The reaction mixture was cooled to rt and the resulting brown precipitate was filtered and purified by silica gel chromatography (EtOAc/PA, 1/1, v/v to 1/0, v/v) to give 6-chloro-8-nitro-2H- benzo[b][l,4]oxazin-3(4H)-one (intermediate 57) (21 g, 78%) as a brown solid.
Intermediate 58:
Figure imgf000096_0001
[0208] Intermediate 57 (326 mg, 1.4 mmol) was dissolved in EtOH, then Pd/C (0.03 g) was added. The mixture was hydrogenated under H2 for 4 h at 50°C. The reaction mixture was filtered over Hyflo and the filtrate was concentrated in vacuo to give 8-amino-2H- benzo[b][l,4]oxazin-3(4H)-one (intermediate 58) (83%) as a brown solid, which was used in the next step without any further purification.
Intermediate 59:
Figure imgf000096_0002
[0209] To a solution of intermediate 58 (164 mg, 1.0 mmol) in chlorobenzene (5 mL) was added bis(2-chloroethyl)amine hydrochloric acid (285 mg, 1.6 mmol) and the mixture was heated to reflux for 100 h. The reaction mixture was concentrated in vacuo and the residue was stirred in EtOAc for 2 h. The resulting brown solid was filtered and purified by silica gel chromatography to give 8-(piperazin-l-yl)-2H-benzo[b][l,4] oxazin-3(4H)-one (intermediate 59) (150 mg, 64%). HPLC: 99%, RT 1.344 min. MS(ESI) m/z 234.1 [M + H]+.
Compound 38:
Figure imgf000096_0003
[0210] A mixture of intermediate 4 (110 mg, 0.37 mmol) and Nal (110 mg, 0.74 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 59 (118 mg, 0.5 mmol) and anhydrous K2C03 (138 mg, 1.92 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 8-(4-(4-(2-oxo- 1,2,3, 4-tetrahydroquinolin-7-yloxy)butyl)piperazin-l-yl)-2H- benzo[b][l,4]oxazin-3(4H)-one (compound 38) (55 mg, 34%). ]H NMR (300 MHz, CDC13): δ 10.56 (s, 1H), 9.96 (s, 1H), 7.04 (d, J= 8.4 Hz, 1H), 6.85 (t, 1H), 6.57-6.43 (m, 4H), 4.53 (s, 2H), 3.92 (t, J = 5.7 Hz, 2H), 2.97 (br, 4H), 2.78 (t, J= 6.9 Hz, 2H), 2.43-2.38 (m, 4H), 1.72-1.58 (m, 4H). HPLC: 99%, RT 1.970 min. MS (ESI) m/z 451.0 [M + H]+.
Synthesis of Compound 39
Scheme 65. Synthesis of compound 39
Figure imgf000097_0001
Compound 39:
Figure imgf000097_0002
[0211] A mixture of intermediate 4 (110 mg, 0.34 mmol) and Nal (102 mg, 1.36 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 55 (117 mg, 0.51 mmol) and anhydrous K2C03 (188 mg, 1.36 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(5-(4-(2-methoxyphenyl)piperazin-l-yl)pentyloxy)-3,4-dihydroquinolin-2(lH)- one (compound 39) (112 mg, 81%). 1H MR (300 MHz, CDC13): δ 7.92 (brs, 1H), 7.06-6.85 (m, 5H), 6.52 (d, J = 6.6 Hz, 1H), 6.31 (s, 1H), 3.98-3.94 (m, 2H), 3.86 (s, 3H), 3.12 (br, 4H), 2.92- 2.87 (m, 2H), 2.69-2.59 (m, 6H), 2.52-2.47 (m, 2H), 1.82-1.72 (m, 4H). HPLC: 99%, RT 2.100 min. MS (ESI) m/z 410.2 [M + H]+. Synthesis of Compound 40
Scheme 66, Synthesis of intermediate 61
Figure imgf000098_0001
Intermediate 60:
Figure imgf000098_0002
[0212] Triethylamine in acetone was added dropwise to the solution of 2,3- dihydrobenzo[b][l,4]dioxine-5-carboxylic acid (360 mg, 2 mmol) in water and acetone at 0°C. To the mixture was added ethyl carbonochloridate in acetone slowly. The mixture was stirred for 4 h then a solution of NaN3 (196 mg, 3.02 mmol) in water was added dropwise. The reaction mixture was stirred for 1 h then it was poured into ice water and extracted with Et20. The combined organic extracts were washed with brine, dried with MgS04, and concentrated in vacuo. The residue was dissolved in anhydrous toluene and the mixture was heated on a steam bath until nitrogen evolution had ceased. The toluene was then removed in vacuo and 20% aq HCI (4 mL) was added and the solution was heated to reflux and stirred overnight. The reaction was concentrated under reduced pressure. The residue was dissolved in water, made strongly alkaline by the addition of 40% aq NaOH, then extracted with Et20. The combined organic extracts were washed with brine, dried by MgS04, and purified by flash chromatography on silica gel column (PE:EtOAc=5:l) to give 2,3-dihydrobenzo[b][l,4]dioxin-5-amine (intermediate 60) (270 mg, 70%). HPLC: 99%, RT 1.214 min. MS (ESI) m/z 152.1 [M + H]+. Intermediate 61:
Figure imgf000099_0001
[0213] Bis(2-chloroethyl)amine hydrochloride salt (452 mg, 2.54 mmol) was added to a solution of intermediate 60 (320 mg, 2.12 mmol) in chlorobenzene (4 mL) and the mixture was heated to reflux and stirred overnight. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on silica gel column (DCM:MeOH=30: 1-20:1), followed by a wash with EtOAc, to give l-(2,3-dihydrobenzo[b][l,4]dioxin-5-yl)piperazine (intermediate 61) (236 mg, 51%). HPLC: 99%, T 1.673 min. MS (ESI) m/z 221.1 [M + H]+.
Compound 40:
Figure imgf000099_0002
[0214] A mixture of intermediate 4 (100 mg, 0.34 mmol) and Nal (102 mg, 0.68 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 61 (112 mg, 0.51 mmol) and anhydrous K2C03 (188 mg, 1.36 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2,3-dihydrobenzo[b][l,4]dioxin-5-yl)piperazin-l-yl)butoxy)-3,4- dihydroquinolin-2(lH)-one (compound 40) (125 mg, 83%). ¾ NMR (300 MHz, CDC13): δ 7.88 (s, 1H), 7.04 (d, J=8.1Hz, 1H), 6.80-6.77(m, 1H), 6.61-6.51 (m, 3H), 6.31 (d, J = 1.8 Hz, 1H), 4.33-4.24 (m, 4H),3.96(m, 2H), 3.10(m, 4H), 2.89 (t, J = 7.5 Hz, 2H), 2.66-2.59 (m, 6H), 2.50- 2.46(m, 2H), 1.81-1.71(m, 6H). HPLC: 99%, RT 2.468 min. MS (ESI) m/z 438.2 [M + H]+.
Synthesis of Compound 41
Scheme 68. S nthesis of com ound 41
Figure imgf000099_0003
xamp e Compound 41:
Figure imgf000100_0001
[0215] A mixture of intermediate 12 (100 mg, 0.32 mmol) and Nal (95 mg, 0.64 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 61 (105 mg, 0.48 mmol) and anhydrous K2C03 (176 mg, 1.27 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2,3-dihydrobenzo[b][l,4]dioxin-5-yl)piperazin-l-yl)butoxy)-3,4-dihydro- l,8-naphthyridin-2(lH)-one (compound 41) (115 mg, 83%). !H MR (300 MHz, CDC13): δ 7.60 (s, 1H), 7.36 (d, J=8.1Hz,lH), 7.27 (s, 1H), 6.78 (t, J= 8.1 Hz, lH),6.61-6.53(m, 2H), 6.35 (d, J = 8.1 Hz, lH),4.33-4.20 (m, 6H), 3.10 (s, 4H), 2.86 (dd, J=7.5 Hz, 2H), 2.67-2.62 (m, 6H),2.51- 2.46 (m, 2H), 1.82-1.69(m, 6H). HPLC: 99%, RT 2.084 min. MS (ESI) m/z 439.2 [M + H]+.
Synthesis of Compound 42
Scheme 69. S nthesis of com ound 42
Figure imgf000100_0002
Compound 42:
Figure imgf000100_0003
[0216] A mixture of intermediate 15 (70 mg, 0.24 mmol) and Nal (70 mg, 0.48 mmol) in CH3CN was heated to reflux for 30 min and then cooled to rt. Intermediate 61 (78 mg, 0.36 mmol) and anhydrous K2C03 (162 mg, 0.96 mmol) were added to the mixture. The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOA. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 30:1) to give 7-(4-(4-(2,3-dihydrobenzo[b][l,4]dioxin-5-yl)piperazin-l-yl)butoxy)quinolin- 2(lH)-one (compound 42) (52 mg, 50%). !H NMR (300 MHz, CDC13): δ 11.65 (brs, 1H), 7.72 (d, J = 9.6 Hz, 1H), 7.45 (d, J = 8.4 Hz, 1H), 6.82-6.75 (m, 3H), 6.61- 6.52 (m, 3H), 4.33-4.26 (m, 4H), 4.12-4.08 (m, 2H), 3.12 (br, 4H), 2.70 (br, 4H), 2.52(br, 2H), 2.04-1.68 (m, 6H). HPLC: 99%, RT 2.167 min. MS (ESI) m/z 436.0 [M + H]+.
Synthesis of Compound 43
Scheme 70. Synthesis of compound 43
Figure imgf000101_0001
Intermediate 62:
Figure imgf000101_0002
[0217] A mixture of 4-(4-bromobutoxy)-l-methyl-2-nitrobenzene (200 mg, 0.7 mmol), intermediate 41 (186 mg, 0.7 mmol) and anhydrous K2C03 (195 mg, 1.4 mmol) was dissolved in CH3CN and the solution was heated to reflux and stirred overnight. The solution was concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 50:1) to give l-(2,3-dichlorophenyl)-4-(4-(4-methyl-3- nitrophenoxy)butyl)piperazine (intermediate 62) (180 mg, 59%) as a yellow solid.
Intermediate 63:
Figure imgf000101_0003
[0218] A mixture of intermediate 62 (180 mg, 0.41 mmol), Fe (70 mg, 1.23 mmol) and ammonium chloride (25 mg, 0.41 mmol) was dissolved in EtOH and ¾0, and the solution was heated to reflux and stirred overnight. The solution was filtered, diluted with water, and then extracted with DCM. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, and concentrated in vacuo to give 5-(4-(4-(2,3- dichlorophenyl)piperazin-l-yl)butoxy)-2-methylaniline (intermediate 63) (150 mg, 90%) as a yellow solid. HPLC: 99%, RT 3.197 min. MS(ESI) m/z 407.9 [M + H]+.
Compound 43:
Figure imgf000102_0001
[0219] A mixture of intermediate 63 (120 mg, 0.29 mmol), acetyl chloride (35 mg, 0.44 mmol) and Et3N (54 mg, 0.54 mmol) was dissolved in DCM and the solution was stirred at rt. The solution was diluted with water and extracted with DCM. The combined organic layers were washed with saturated aq NaHC03, brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by flash chromatography on silica gel column (elution with DCM/MeOH = 100:1- 30: 1) to give N-(5-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)-2-methylphenyl)acetamide (compound 43) (45 mg, 35%) as a yellow solid. ¾ NMR (300 MHz, CDC13): δ 7.56 (s, 1H), 7.18-7.13 (m, 2H), 7.07-6.88 (m, 3H), 6.62 (d, J= 8.1 Hz, 1H), 3.99 (br, 2H), 3.26 (br, 4H), 2.92- 2.75 (br, 6H), 2.21 (s, 3H), 2.19 (s, 3H), 1.86 (br, 6H). HPLC: 99%, RT 2.464 min. MS (ESI) m/z 449.8 [M + H]+.
Synthesis of Compound 44
Scheme 71. Synthesis of compound 44
Figure imgf000102_0002
[0220] A mixture of intermediate 44 (41 mg, 0.16 mmol) and Nal (48.1 mg, 0.32 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 47 (64.3 mg, 0.24 mmol), followed then by DIEA (0.062 mL, 0.35 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 6-(3-(4-(2,3-dichlorophenyl)piperidin-l- yl)propoxy)-lH-indazole (compound 44) (42, 1 mg, 65%). 1H NMR (400 MHz, CDC13): δ 7.92 (d, J = 0.8 Hz, 1H), 7.57 (d, J = 8.8 Hz, 1H), 7.33 (dd, J = 7.8, 1.6 Hz, 1H), 7.28 - 7.24 (dd, J = 7.8, 1 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 6.98 (s, 1H), 6.77 (dd, J = 8.8, 2.1 Hz, 1H), 4.10 (t, J = 6.0 Hz, 2H), 3.56 - 3.40 (m, 2H), 3.39 - 3.26 (m, 1H), 3.11 - 2.97 (m, 2H), 2.73 (s, 2H), 2.39 - 2.15 (m, 4H), 2.04 - 1.94 (dd, J = 16.8, 9.4 Hz, 2H). HPLC: 99%, RT 4.397 min. MS (ESI) m/z 404.15 [M + H]+.
Synthesis of Compound 45
Scheme 72. Synthesis of compound 45
Figure imgf000103_0001
[0221] A mixture of intermediate 51 (40.6 mg, 0.15 mmol) and Nal (45.2 mg, 0.30 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 47 (60.3 mg, 0.23 mmol), followed then by DIEA (0.058 mL, 0.33 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 6-(4-(4-(2,3- dichlorophenyl)piperidin-l-yl)butoxy)-lH-indazole (compound 45) (37.6 mg, 60%). lH NMR (400 MHz, CDC13): δ 7.93 (s, 1H), 7.54 (d, J= 8.8 Hz, 1H), 7.33 (dd, J= 7.9, 1.2 Hz, 1H), 7.25 - 7.20 (m, 1H), 7.15 (t, J= 7.9 Hz, 1H), 6.93 (s, 1H), 6.74 (dd, J = 8.8, 1.9 Hz, 1H), 3.94 (t, J = 6.0 Hz, 2H), 3.75 - 3.67 (m, 2H), 3.46 - 3.36 (m, 1H), 3.15 - 3.08 (m, 2H), 3.03 - 2.91 (m, 2H), 2.59 - 2.43 (m, 2H), 2.18 - 2.06 (m, 2H), 2.06 - 1.96 (m, 2H), 1.88 - 1.75 (m, 2H). HPLC: 99%, RT 4.447 min. MS (ESI) m/z 418.10 [M + H]+. Synthesis of Compound 46
Scheme 73. Synthesis of compound 46
Figure imgf000104_0001
[0222] A mixture of intermediate 44 (43.7 mg, 0.17 mmol) and Nal (51.3 mg, 0.34 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 2 (57.9 mg, 0.21 mmol), followed then by DIEA (0.066 mL, 0.38 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 6-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)propoxy)-lH-indazole (compound 46) (43.6 mg, 61%). !H NMR (400 MHz, CDC13): δ 7.96 (d, J= 0.8 Hz, 1H), 7.59 (d, J= 8.8 Hz, 1H), 7.15 - 7.07 (m, 2H), 7.00 (dd, J= 7.4, 2.3 Hz, 1H), 6.88 (s, 1H), 6.80 (dd, J= 8.8, 2.1 Hz, 1H), 4.09 (t, J= 6.1 Hz, 2H), 3.43 - 3.37 (m, 2H), 3.29 (t, J= 6.1 Hz, 2H), 3.14 - 3.05 (m, 4H), 3.00 - 2.92 (m, 2H), 2.22 - 2.10 (m, 4H). HPLC: 99%, RT 4.201 min. MS(ESI) m/z 419.10 [M + H]+.
Synthesis of Compound 47
Scheme 74. Synthesis of compound 47
Figure imgf000104_0002
Compound 47:
Figure imgf000105_0001
[0223] A mixture of intermediate 51 (54.2 mg, 0.20 mmol) and Nal (60.3 mg, 0.40 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 2 (62.4 mg, 0.22 mmol), followed then by DIEA (0.077 mL, 0.44 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 6-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l- yl)butoxy)-lH-indazole (compound 47) (50.8 mg, 59%). HPLC: 99%, RT 4.526 min. MS(ESI) m/z 433.20 [M + H]+.
Synthesis of Compound 48
Scheme 75. Synthesis of compound 48
Figure imgf000105_0002
Intermediate 44 Intermediate 64 CH3CN Example 48
Compound 48:
Figure imgf000105_0003
[0224] A mixture of intermediate 44 (43.6 mg, 0.17 mmol) and Nal (51.2 mg, 0.34 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 64 (45.0 mg, 0.21 mmol, prepared following the procedure in US Publication No. 2010/0119622) followed by DIEA (0.066 mL, 0.38 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 7-(4-(3-(lH-indazol-6-yloxy)propyl)piperazin-l- yl)benzo[d]oxazol-2(3H)-one (compound 48) (43.7 mg, 65%). HPLC: 99%, RT 3.551 min. MS (ESI) m/z 394.20 [M + H]+.
Synthesis of Compound 49
Scheme 76. Synthesis of compound 49
Figure imgf000106_0001
Compound 49:
Figure imgf000106_0002
[0225] A mixture of intermediate 51 (47.3 mg, 0.18 mmol) and Nal (52.6 mg, 0.35 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 64 (42.4 mg, 0.19 mmol), followed then by DIEA (0.067 mL, 0.39 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 7-(4-(4-(lH-indazol-6- yloxy)butyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one (compound 49) (43.5 mg, 61%). HPLC: 99%, RT 3.680 min. MS(ESI) m/z 408.20 [M + H]+.
Synthesis of Compound 50
Scheme 77. Synthesis of intermediate 65
Figure imgf000106_0003
diglyme Intermediate 65 Scheme 78. Synthesis of compound 50
Figure imgf000107_0001
Intermediate 44 Intermediate 65 CH3CN Example 50
Intermediate 65:
Figure imgf000107_0002
[0226] To a solution of l-bromo-2-methoxybenzene (0.623 mL, 5.0 mmol) and piperazine (1722.8 mg, 20.0 mmol) in diglyme (5 mL) was added t-BuONa. The resulting mixture was stirred at rt for a few minutes before the addition of dichlorobis(tri-0- tolyphosphine)palladium (235.8 mg, 0.3 mmol). The resulting mixture was then stirred at 170°C under microwave irradiation for 30 min. Water was added and the mixture was extracted with EtOAc. The combined organic layers were dried, concentrated, and purified by ISCO to give 1- (2-methoxyphenyl)piperazine (intermediate 65) (758 mg, 79%) as an off-white solid. HPLC: 95%, RT 2.641 min. MS (ESI) m/z 193.10 [M + H]+.
Compound 50:
Figure imgf000107_0003
[0227] A mixture of intermediate 44 (42.2 mg, 0.17 mmol) and Nal (49.5 mg, 0.33 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 65 (35.0 mg, 0.18 mmol), followed then by DIEA (0.063 mL, 0.36 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 6-(3-(4-(2- methoxyphenyl)piperazin-l-yl)propoxy)-lH-indazole (compound 50) (36.6 mg, 60%). HPLC: 99%, RT 3.907 min. MS(ESI) m/z 367.20 [M + H]+. Synthesis of Compound 51
Scheme 79. Synthesis of compound 51
Figure imgf000108_0001
Compound 51:
Figure imgf000108_0002
[0228] A mixture of intermediate 51 (41.6 mg, 0.15 mmol) and Nal (46.3 mg, 0.31 mmol) in CH3CN (3 mL) was heated to reflux for 30 min and then cooled to rt. To this mixture was added intermediate 65 (32.7 mg, 0.17 mmol), followed then by DIEA (0.059 mL, 0.34 mmol). The resulting mixture was heated to reflux and stirred for 4 h. Precipitated crystals were filtered off and the filtrate was evaporated under reduced pressure. The residue was extracted with EtOAc. The combined EtOAc layers were washed with brine, dried over anhydrous Na2S04, concentrated in vacuo and purified by ISCO to give 6-(4-(4-(2- methoxyphenyl)piperazin-l-yl)butoxy)-lH-indazole (compound 51) (35.6 mg, 62%). HPLC: 99%, RT 4.000 min. MS (ESI) m/z 381.20 [M + H]+.
EXAMPLE 2
In Vitro Studies
[0229] Each of the synthesized compounds were tested in a variety of in vitro assays for D2 receptor activity. Results are summarized in Table 6.
CHO-D? Membrane Preparation for GTP-gamma-S and Radioligand Binding Assays
[0230] Cells stably expressing D2 receptors (CHO-D2) were plated in 15-cm dishes (in DMEM containing 10% fetal bovine serum and grown to 90% confluence. Then, cells were washed with PBS, pH 7.4, and harvested by scraping into PBS, pH 7.4. Harvested cells were centrifuged at 1,000 x g for 10 min, then hypotonically lysed by resuspension in ice-cold 50 mM HEPES, 1% BSA, pH 7.4. Membranes were isolated by centrifugation at 21,000 x g for 20 min. The supernatant was removed and the membrane pellets were assayed for D2 ligand-stimulated GTP-gamma-S binding or stored at -80°C until used for radioligand binding assays.
CHO-D? Radioligand Binding Assay
[0231] Membranes prepared as above were resuspended to 1 μg protein/μΐ (measured by Bradford assay using BSA as standard), and 50 μΐ were added to each well of a polypropylene 96-well plate containing (per well): 50 μΐ of buffer (20 mM HEPES, 10 mM MgCl2, 1 mM EDTA, 1 mM EGTA, 100 mM N-methyl-D-gluconate, pH 7.4), 50 μΐ of 1.5 nM [3H]N- methylspiperone (final concentration 0.3 nM) and reference or D2 test ligand at various concentrations ranging from 50 pM to 50 μΜ (final concentrations ranging from 10 pM to 10 μΜ, triplicate determinations for each concentration of D2 test ligand). After a 1.5-hr incubation in the dark at room temperature, the reactions were harvested onto 0.3% PEI-soaked Filtermax GF/A filters (Wallac) and washed three times with ice-cold 50 mM Tris, pH 7.4 using a Perkin- Elmer Filtermate 96-well harvester. The filters were subsequently dried, placed on a hot plate (100°C), and Melitilex-A (Wallac) scintillant was applied. The filters were then removed from the hot plate and allowed to cool. The filters were counted on a Wallac TriLux microbeta counter (3 min per well). Residual [3H]N-methylspiperone binding to filtered membranes was plotted as a function of log [reference] or log [D2 test ligand] and the data were regressed using the one-site competition model built into Prism 4.0 (GraphPad software).
D? mediated cAMP GloSensor Assay
[0232] HEK293T cells co-expressing the cAMP biosensor GloSensor-22F (Promega) and hD2 receptors were seeded (10,000 cells/20 μΐ/well) into white, clear-bottom, tissue culture plates in HBSS, 10% FBS, 20 mM HEPES, pH 7.4. After a one- to two-hour recoveiy, cells were treated with 10 μΐ of 3X test or reference drug prepared in HBSS, 10% FBS, 20 mM HEPES, pH 7.4. After 30 minutes, cells were challenged with 10 μΐ of 1,200 nM (4X) isoproterenol in 8% (4X) GloSensor reagent. Luminescence per well per second was read on a Wallac TriLux microbeta plate counter. Data were normalized to the isoproterenol response (100%) and the maximal quinpirole-induced inhibition thereof (0%) and regressed using the sigmoidal dose- response function built into GraphPad Prism 4.0. Notably, HEK293T cells expressing the GloSensor-22F alone (no hD2) were assayed in parallel and displayed no inhibition of isoproterenol-stimulated cAMP, either by quinpirole or by the test compounds, suggesting that the effect observed in hD2-expressing cells was due to compound acting via the recombinant receptor.
D? β-Arrestin Recruitment (Tango) assay
[0233] Recruitment of β-arrestin to agonist-stimulated D2 receptors was performed using a previously described "Tango"-type assay (Barnea et at, Proc. Natl. Acad. Sci. USA 105:64 (2008)). Briefly, HTLA cells stably expressing β-arrestin-TEV protease and a tetracycline transactivator-driven luciferase were plated in 15-cm dishes in DMEM containing 10% fetal bovine serum and transfected (via calcium phosphate) with 20 μg of a D2V2-TCS-tTA construct (Barnea et al, Proc. Natl. Acad. Sci. USA 105:64 (2008)). The next day, cells were plated in white, clear-bottom, 384-well plates (Greiner, 10,000 cells/well, 50 μΐ/well) in DMEM containing 1% dialyzed fetal bovine serum. The following day, the cells were challenged with 10 μΐ/well of reference agonist (6 μΜ) or D2 test ligand (6 μΜ) ± reference agonist prepared in HBS, 20 mM HEPES, pH 7.4, 18% DMSO (final ligand concentrations were 1 μΜ, final DMSO concentration was 3%). After 18 hours, the medium was removed and replaced with IX BriteGlo reagent (Promega), and luminescence/well was read using a TriLux plate reader (1 sec/well). Data were normalized to vehicle (0%) and quinpirole (100%) controls and regressed using the sigmoidal dose-response function built into GraphPad Prism 4.0.
pERK High-Content Assay
[0234] Cell Culture: Chinese Hamster Ovary (CHO) cells stably expressing the hD2L dopamine receptor (Urban et al, Neuropsychopharmacology 32:67 (2007)) were maintained in Ham's F12 medium supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 μg/mL streptomycin, and 0.5 μg/ml G418. On day 1 of the assay, cells were seeded onto black clear-bottom tissue culture-treated 96-well plates (Greiner, BioOne). On day 2 of the assay, cells were washed with serum-free medium (Ham's F-12, penicillin and streptomycin) and incubated in 100 μΐ serum-free medium overnight .
[0235] Immunofluorescence: Automatic multichannel pipetters were used for liquid handling and multichannel vacuum manifolds for aspirations. Each tested concentration was typically measured in triplicate or quadruplicate. For concentration curves, half-log-dilutions were used. Drug dilutions were prepared in stimulation medium (serum-free medium, 100 mg/L ascorbic acid). 100 μΜ dopamine and 1 μΜ PMA (phorbol 12-myristate 13 -acetate) were used as positive controls. Medium only was used as a negative control. Specificity of the response was confirmed by pretreatment for 5 min with 10 μΜ of the antagonist spiperone and in experiments using wild type CHO cells.
[0236] Cells were stimulated on day 3 by fast addition of 50 μΐ^ equilibrated 3X drug dilutions in a tissue culture incubator for 5 min. Plates were placed on ice, the medium was aspirated, and 100 μΕΛνεΙΙ of freshly prepared ice-cold fixing buffer (4% formaldehyde and 0.5 mM CaCl2 in PBS) solution was added. After 30 min fixing at room temperature, the plates were washed with 350 μΐ ννεΐΐ PBS/Ca (0.5 mM CaCl2 in PBS) and permeabilized for 20 min in 100 μΙ,ΛνεΙΙ 0.3% Triton X-100 in PBS/Ca. Plates were incubated in 100 μΙ,ΛνεΙΙ blocking buffer (PBS/Ca, 5% goat serum, 0.1% Triton X-100) for 1 h at room temperature and then in blocking buffer containing rabbit phospho-Thr202/p-Tyr204-ERK antibody (Cell Signaling 9101) at 1 :1000 at 4°C overnight. Cells were washed 3X for 5-10 min with 250 μΕΛνεΙΙ wash buffer (PBS/Ca, 0.03% Triton X-100). Plates were incubated with 50 μΕΛνεΙΙ Alexa-594 coupled goat anti-rabbit IgG (Invitrogen) at 1 :250, 5 μg/mL Hoechst 33342 nuclear stain, and 25 μg/mL concanavaline-Alexa488 conjugate (ConA, Invitrogen) in blocking buffer for 2 h at room temperature. Plates were washed three times with 250 μΙ^ΛνεΙΙ washing buffer, post-fixed for 10 min in fixing buffer, washed with 250 μΐ PBS/Ca, filled with 200 nL/well PBS/Ca, sealed with transparent adhesive plate seals, and stored at 4°C.
[0237] Microscopy and Image Analysis: Plates were scanned with a high-content automated microscopic system (BD Pathway 855) using a 20x objective and a 2 x 2 image montage setting. The Alexa594 light path was used for the target signal, Alexa488 for whole cell staining, and Hoechst for the nuclear staining. Images were analyzed using the CellProfiler software. Well-averaged individual cell-based measurements were exported to Excel and cell- free background intensity was subtracted from the whole cell intensity. Concentration curves were analyzed in GraphPad Prism by fitting against a sigmoidal dose-response model and normalization to the dopamine curve.
[0238] The results in Table 6 show that, of the 43 compounds tested, 33 are partial agonists (Emax of 10% - 80%) of the β-arrestin pathway and another 8 compounds are full agonists (Emax > 80%) of the β-arrestin pathway. The compounds are also exceedingly potent, with 39 having an EC5o less than 100 nM and 23 having an EC50 less than 10 nM.
[0239] The cAMP (Glo-Sensor) assay shows that the majority of the tested compounds are not agonists for the Gj-coupled signaling pathway and that 14 of the 43 compounds function as inverse agonists for the Gj-coupled signaling pathway (i.e., have a negative Emax). The p-ERK assay confirms that the majority of compounds lack agonism via canonical G-protein-dependent pathways.
[0240] Compounds 2 and 3 were tested for receptor binding specificity. Consistent with their high potency in D2 functional assays, 2 and 3 displayed high affinities (Kfs < 10 nM) in the D2 radioligand competition binding assay (Table 7). Although 2 and 3 also had high affinity for D3-dopamine receptor, they displayed low affinities for other dopamine receptors (i.e., Dl5 D4, and D5). At serotonin receptors, 2 and 3 had moderate to high binding affinity ( j's: 0.6 - 250 nM) at 5HT2A, 5HT2B, 5HT2C, and 5HTiA, but were significantly less potent in functional assays (Ca2+ mobilization fluorometric imaging plate reader (FLIPR) or cAMP biosensor). 2 and 3 were antagonists at 5HT2A, 5HT2B, and 5HT2C and weak agonists (EC50's > 1 μΜ) at 5HTiA. In addition, 2 and 3 had moderate affinities to Hi -histamine receptor (Xj's < 10 nM) but were less potent in Hi functional assays. In general, 2 and 3 display a similar G protein-coupled receptor selectivity profile as aripiprazole (Table 7). ATTORNEY DOCKET NO. 5470-568WO
Table 6
Figure imgf000112_0001
ATTORNEY DOCKET NO. 5470-568WO
Figure imgf000113_0001
Neutral ligands are either antagonists or are inactive; NA: not applicable.
I l l
Table 7
Figure imgf000114_0001
* Kj, IC50, pA2, or EC50 values are the average of at least two duplicate experiments with standard deviation (SD) values that are 3-fold less than the average.
[0241] In mouse pharmacokinetic (PK) studies, both 3 and aripiprazole displayed high exposure levels in brain and excellent CNS penetration (Table 8). Although the brain exposure level of 3 was about 3-fold lower, 3 had a longer half life in brain and higher brain plasma ratio over 24 h compared to aripiprazole.
Table 8
Figure imgf000115_0001
EXAMPLE 3
In Vivo Studies
[0242] Compounds 2 (UNC10000006A, or UNC0006) and 3 (UNC10099975A, or UNC9975) were evaluated in D-amphetamine-induced hyperlocomotion, phencyclidine (PCP)- induced hyperlocomotion, and/or catalepsy mouse models.
[0243] Mice. All experiments were approved by the Institutional Animal Care and Use Committees at the University of North Carolina, Chapel Hill and Duke University. C57BL/6J and P-arrestin-2 knockout (Bohn et al, Science 286:2495 (1999)) mice were housed under standard conditions - 12 hour light/dark cycle and food and water provided ad libitum. Adult, age-matched male and female C57BL/6J and P-arrestin-2 knockout drug naive mice were used for all behavioral testing.
[0244] Locomotor activity. Mouse locomotor activity was assessed in photocell-based activity chambers under standardized environmental conditions, using an AccuScan activity monitor (AccuScan Instruments, Columbus, OH) with a 25.8 x 25.8 cm Plexiglas chamber and a beam spacing of 1.52 cm as described (Abbas et al, J. Neurosci. 29:7124 (2009)). Mice were injected (i.p.) with vehicle (0.9% saline/0.2% acetic acid) , aripiprazole (0.10, 0.25, 0.50 or 2.0 mg/kg), or UNC9975 (0.25, 0.5 and 2.0 mg/kg) and placed into the open field. Thirty min later D-amphetamine (3 mg/kg) or phencyclidine (6.0 mg/kg) was administered and they were immediately returned to the open field for 80 min. Activity was monitored throughout this entire period . Horizontal activity was measured as the total distance traveled in centimeters. The means ± SEMs of the locomotor responses of animals were analyzed using Graphpad Prism 5.0. To estimate the half-maximal inhibitory concentration (ED50), dose responses of total locomotor activity during the 90 minutes after D-amphetamine or phencyclidine administration were plotted and best-fit decay curves were determined using a nonlinear regression one-phase decay equation. Cumulative locomotor responses were analyzed using a one-way ANOVA followed by Newman- Keuls multiple comparison test using Graphpad Prism 5.0. Statistical significance was set at p<0.05.
[0245] Catalepsy testing. In this testing paradigm, mice were initially injected (i.p.) with vehicle (0.9% saline/0.2% acetic acid), aripiprazole, U C0006 (5 mg/kg), UNC9975 (5 mg/kg) or haloperidol (2 mg/kg) and returned to their home cage. The latency of movement was assessed 30, 60, 90 and 120 min after drug injection; the maximal latency to move in the mice was observed 60 min after drug treatments. Mice were placed upright on a screen placed at a 45 degree angle. The time required for the animal to move all four paws was scored in seconds (maximum of 5 minutes) and is reported as the latency to movement. An extended delay to voluntarily move on the inclined screen test is indicative of drug-induced catalepsy. The data are displayed as mean ± SEM. Catalepsy data were analyzed for statistical significance using a oneway ANOVA followed by Newman-Keuls multiple comparison test using Graphpad Prism 5.0. Differences were considered significant at p<0.05.
[0246] UNC9975 exhibited potent antipsychotic-like activity in mouse hyperlocomotion studies which is attenuated in p-arrestin-2 knockout mice . In Fig. 1A, locomotion responses in inbred C57BL/6 mice are shown as 5 minute binned intervals to different doses (i.p.) of UNC9975 or vehicle followed 30 min later by 3 mg/kg D-Amphetamine (AMPH, i.p.). In Fig. IB, total distance traveled after D-Amphetamine administration (30-70 min time interval) is shown. C57BL/6 mice were given vehicle or different doses of UNC9975 or aripiprazole 30 min prior to AMPH treatment. n=8 animals/group. *, p<0.05 versus vehicle + 3 mg/kg D- Amphetamine group.
[0247] In Figs. 1C and ID, locomotor activities are shown as 5 min binned intervals of wild-type (WT) or P-arrestin-2 knockout (β-ΑΓΓ<2 KO) littermate mice given vehicle or different doses of UNC9975 followed 30 min later with 6 mg/kg phencyclidine (PCP, i.p.). Fig. IE shows the total distance traveled by WT and P-ARR2 KO mice after PCP administration (30 to 70 min time interval), shown in Figs. 1C and ID - different doses of UNC9975 were given to mice 30 min before PCP treatment, n = 14 WT and P-ARR2 KO pairs/group. *p<0.05, versus vehicle + 6 mg/kg PCP group. Fig. IF shows the total distance traveled by WT and -ARR2 KO mice after aripiprazole followed by PCP treatments (30 to 70 min time interval), n = 8 littermate WT and β- ARR2 KO pairs/group. *p<0.05, versus vehicle + 6 mg/kg PCP group.
[0248] UNC9975 and U C0006 induced catalepsy in p-arrestin-2 knockout mice but not in wild-type littermates. In Figs. 2A and 2B, wild-type and p-Arrestin-2 knockout littermate mice were administered (i.p.) vehicle, 5.0 mg/kg UNC9975, 5.0 mg/kg UNC0006, 5.0 mg/kg aripiprazole, or 2.0 mg/kg haloperidol. Following 30 and 60 min after drug injection, catalepsy was assessed using the inclined screen test where latency to move was scored, n = 8 littermate wild-type and knockout animal pairs/group. *p<0.05 versus vehicle controls.
[0249] Through a combined medicinal chemistry and comprehensive in vitro and in vivo pharmacological profiling approach, novel D2 β-arrestin-biased agonists were designed, synthesized and characterized, and demonstrated to have unique atypical antipsychotic drug-like activities in vivo. These novel compounds represent the first functionally selective β-arrestin- biased dopamine D2 receptor ligands to exhibit antipsychotic activity in vivo. These findings show that based on the arrestin-bias of these compounds, β-arrestin may be an important contributor to both antipsychotic drug efficacy and antipsychotic side effects.
[0250] The disclosed compounds were discovered as unique, β-arrestin-biased functionally selective D2 ligands. In addition to possessing high affinity for D2 receptors, both UNC0006 and UNC9975 are potent partial agonists which induce D2 receptors-mediated β- arrestin recruitment and signaling, and simultaneously are potent inverse agonists at Gi-dependent signaling. More significantly, the in vivo atypical antipsychotic drug-like activity of UNC9975 indicated a strict requirement for p-arrestin-2 for both full antipsychotic activity and protection against motoric side-effects. Similar to aripiprazole, this β-arrestin-biased ligand shows a potent ability to suppress both D-amphetamine and phencyclidine-induced hyperlocomotion in mice, indicating that the compound possesses antipsychotic drug-like activities in vivo. Significantly, the antipsychotic drug-like activity of UNC9975 was attenuated in β-3ΓΓε8ΐϊη-2 knockout mice, indicating p-arrestin-2 is required in vivo for full activity.
[0251] With the exception of aripiprazole, all FDA approved typical and atypical antipsychotic medications (e.g., haloperidol, chlorpromazine, clozapine, risperidone) share the common property of antagonizing D2-mediated G protein-dependent and independent signaling (Masri et al, Proc. Natl. Acad. Sci. USA 105:13656 (2008)). Indeed, typical and atypical antipsychotic drugs, with the exception of aripiprazole, are inverse agonists at Gj-mediated signaling and antagonists at arrestin-ergic pathways. This combination of inverse agonism and antagonism is thought to underlie both the therapeutic benefit to prevent psychotic symptoms but can also cause serious extrapyramidal side-effects including catalepsy and other motor diskenesias (Roth et al, Nat. Rev. Drug Discov. 3:353 (2004)). Although aripiprazole, UNC0006 and UNC9975 do not induce catalepsy in wild-type mice at doses at which antipsychotic druglike therapeutic activities are measured, UNC0006 and UNC9975 resemble haloperidol in inducing catalepsy in p-arrestin-2 knockout mice. This propensity to cause catalepsy in the absence of P-arrestin-2 suggests that UNC0006 and UNC9975 signal through P-arrestin-2 in vivo and that this signaling may protect against motoric side-effects due to inverse agonism at D2 receptor Gj pathways.
[0252] These findings have obvious implications for the development of novel therapeutic approaches for treating schizophrenia and related disorders. Atypical antipsychotic drugs, which differ from older medications {e.g. , haloperidol and chlorpromazine) by virtue of their reduced propensity to induce motoric side-effects, are among the most widely prescribed medications. Many pharmacologic strategies which aim to target a multiplicity of non-D2 receptor molecular targets {e.g., 5-HT2A inverse agonists, 5-HT2C agonists, mGluR2/3 agonists, NK-3 antagonists, sigma antagonists, Dp or D4-selective antagonists and so forth) have led to a growing number of unsuccessful attempts to create safer and more effective atypical antipsychotic drugs (Roth et al, Nat. Rev. Drug Discov. 3:353 (2004); Conn et al, Neuropsychopharmacology 33:2048 (2008); Gray et al, Mol. Psychiatry 12:904 (2007)). The present results suggest that novel atypical antipsychotic drugs with a unique mechanism of action may arise from the disclosed β-arrestin-biased D2 ligands. These compounds may demonstrate special efficacies in psychotic disorders with a mood dysfunction given the prominent role of arrestin-ergic signaling in the action of lithium and related compounds.
[0253] The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein. All publications, patent applications, patents, patent publications, and any other references cited herein are incorporated by reference in their entireties for the teachings relevant to the sentence and/or paragraph in which the reference is presented.

Claims

What is Claimed is:
1. A compound of formula I:
Figure imgf000119_0001
wherein m is 0 or 1;
n is 0, 1, or 2;
q is 1 or 2;
X is O, NH, or C¾;
Y is N or CH;
Z is C, CH, C¾, cycloalkyl, aryl, or heteroaryl;
is a single, double, or triple bond as valencies permit; and
Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
2. The compound of claim 1, which is β-arrestin biased.
3. The compound of claim 1, having the structure of formula II:
Figure imgf000119_0002
wherein n is 0, 1 , or 2;
q is 1 or 2;
X is O, NH, or CH2;
Y is N or CH; Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is a single, double, or triple bond as valencies permit; and
Art and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
4. The compound of claim 3, having the structure of formula III:
Figure imgf000120_0001
wherein n is 0, 1, or 2;
X is O, NH, or CH2;
Y is N or CH; and
Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof. compound of claim 3, having the structure of formula IV
Figure imgf000120_0002
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
6. The compound of claim 3, wherein n is 1.
7. The compound of claim 3, wherein X is O.
8. The compound of claim 3 wherein Ar2 is:
Figure imgf000121_0001
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof. 10. The compound of claim 3, selected from the group consisting of:
Figure imgf000121_0002
Figure imgf000122_0001
Figure imgf000122_0002
120
11. The compound of claim 3, selected from the group consisting of:
Figure imgf000123_0001
Figure imgf000124_0001
Figure imgf000125_0001
Figure imgf000125_0002
Figure imgf000125_0003
12. The compound of claim 3, selected from the group consisting of:
-(3-(4-(2,3-dichloiOphenyl)-l,4-diazepan-l -yl)propoxy)-3,4-dihydroquinolin-2(lH)-one;
-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one;
-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;-(4-(4-(2,3-dichlorophenyl)-l ,4-diazepan-l-yl)butoxy)quinolin-2(lH)-one;
-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)propoxy)-3,4-dihydroisoquinolin-l(2H)-one;-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydroisoquinolin-l (2H)-one; 5-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butoxy)benzo[< ]thiazole;
5-(3-(4-(2,3-dichlorophenyl)-l ,4-diazepan-l-yl)propoxy)benzo[ ]thiazole;
7-(4-(4-(2-methoxyphenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one;
7-(4-(4-(2-methoxyphenyl)-l,4-diazepan-l-yl)butoxy)quinolin-2(lH)-one;
7-(4-(4-(2-methoxyphenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(2-ethoxyphenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one;
7-(4-(4-(2-ethoxyphenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(2-ethoxyphenyl)- 1 ,4-diazepan- 1 -yl)butoxy)quinolin-2(lH)-one;
7-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l -yl)propoxy)quinolin-2(lH)-one;
7-(4-(4-(2-isopropoxyphenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one;
7-(4-(4-(2-isopropoxyphenyl)-l ,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)- one;
7-(4-(4-(2-isopropoxyphenyl)-l,4-diazepan-l-yl)butoxy)quinolin-2(lH)-one;
5- (3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)propoxy)-lH-benzo[^imidazol-2(3H)-one;
6- (3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)propoxy)-lH-indazole; and
6- (4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butoxy)-lH-indazole.
13. The compound of claim 3, selected from the group consisting of:
7- (4-(4-(2-oxo-2,3-dihydro-lH-benzo[d]imidazol-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro- 1 ,8-naphthyridin-2(lH)-one;
7-(4-(4-(7-oxo-5,6,7,8-tetrahydro-l,8-naphthyridin-2-yloxy)butyl)-l,4-diazepan-l- yl)benzo[d]oxazol-2(3H)-one;
4-(4-(4-(7-oxo-5,6,7,8-tetrahydro-l,8-naphthyridin-2-yloxy)butyl)-l,4-diazepan-l- yl)benzo[d]oxazol-2(3H)-one;
7-(4-(4-(lH-benzo[d]imidazol-7-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin- 2(lH)-one;
7-(4-(4-(benzo[d]oxazol-7-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)- one;
7-(4-(4-(benzo[d]oxazol-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)- one;
7-(4-(4-(lH-indazol-7-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one; 7-(4-(4-(lH-indol-7-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one; 7-(4-(4-(benzofuran-7-yl)- 1 ,4-diazepan- 1 -yl)butoxy)-3 ,4-dihydro- 1 , 8-naphthyridin-2(l H)-one; 7-(4-(4-(benzo[b]thiophen-7-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)- one; 7-(4-(4-(benzo[d]thiazol-7-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)- one;
7-(4-(4-(benzo[d]thiazol-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)- one;
7-(4-(4-( 1 H-indol-4-yl)- 1 ,4-diazepan- 1 -yl)butoxy)-3 ,4-dihydro- 1 , 8-naphthyridin-2( 1 H)-one; 7-(4-(4-(lH-indazol-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one; 7-(4-(4-(benzofuran-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one; 7-(4-(4-(benzo[b]thiophen-4-yl)- 1 ,4-diazepan- 1 -yl)butoxy)-3 ,4-dihydro- 1 , 8-naphthyridin-2( 1 H)- one;
7-(4-(4-(naphthalen- 1 -yl)- 1 ,4-diazepan- 1 -yl)butoxy)-3 ,4-dihydro- 1 ,8-naphthyridin-2( 1 H)-one;
7-(4-(4-(isoquinolin-l-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(quinazolin-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(isoquinolin-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(quinolin-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(quinazolin-5-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(quinazolin-8-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(quinolin-5-yl)- 1 ,4-diazepan- 1 -yl)butoxy)-3,4-dihydro- 1 ,8-naphthyridin-2(lH)-one;
7-(4-(4-(isoquinolin-5-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(isoquinolin-8-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(quinolin-8-yl)- 1 ,4-diazepan- 1 -yl)butoxy)-3,4-dihydro- 1 ,8-naphthyridin-2(lH)-one;
7-(4-(4-phenyl-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(pyridin-2-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(pyridin-3-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(pyridin-4-yl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(2-fluorophenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(2-chlorophenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
7-(4-(4-(2-(trifluoromethyl)phenyl)-l,4-diazepan-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one;
2-(4-(4-(7-oxo-5,6,7,8-tetrahydro- 1 ,8-naphthyridin-2-yloxy)butyl)- 1 ,4-diazepan- 1 - yl)benzonitrile;
7-(4-(4-o-tolyl-l,4-diazepan-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
N-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)propyl)-lH-indazol-6-amine;
N-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butyl)-lH-indazol-6-amine;
N-(3-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)propyl)benzo[d]thiazol-5-amine;
N-(4-(4-(2,3-dichlorophenyl)- 1 ,4-diazepan- 1 -yl)butyl)benzo[d]thiazol-5-amine;
5-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butoxy)-lH-benzo[d]imidazol-2(3H)-one; 5 -(3 -(4-(2,3 -dichlorophenyl)- 1 ,4-diazepan- 1 -yl)propylamino)- 1 H-benzo [d] imidazol-2(3H)-one; and
5-(4-(4-(2,3-dichlorophenyl)-l,4-diazepan-l-yl)butylamino)-lH-benzo[d]imidazol-2(3H)-one. 14. The compound of claim 1 , having the structure of formula VI:
Figure imgf000128_0001
wherein n is 0, 1, or 2;
q is 1 or 2;
X is O, H, or CH2;
Y is N or CH;
Z is C, CH, CH2, cycloalkyl, aryl, or heteroaryl;
is a single, double, or triple bond as valencies permit; and
Ari and Ar2 are independently an unsubstituted or substituted monocyclic or bicyclic aryl or heteroaryl;
or a pharmaceutically acceptable salt, prodrug, or optical isomer thereof.
15. The compound of claim 14, wherein n is 0 or 1.
16. The compound of claim 14, wherein X is O or NH.
17. The compound of claim 14, wherein Ari is dichlorophenyl.
18. The compound of claim 14, wherein Ar2 is selected from the group consisting of lH-indazole, benzo[d]thiazole, and imidazol-2(3H)-one.
19. The compound of claim 14, wherein Ar2 is not naphthyridinone, dihydronaphthyridinone, quinolinone, dihydroquinolinone, tetrahydropyridoazepinone, or isoindole.
20. The compound of claims 14, selected from the group consisting of:
Figure imgf000129_0001
Figure imgf000129_0002
Figure imgf000130_0001
128
Figure imgf000131_0001
Figure imgf000131_0002
Figure imgf000132_0001
Figure imgf000132_0002
130
Figure imgf000133_0001
Figure imgf000133_0002
22. The compound of claim 14, selected from the group consisting of:
6- (3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)-lH-indazole;
5-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butoxy)benzo[d]thiazole;
5-(3-(4-(2,3-dichlorophenyl)piperidin-l-yl)propoxy)benzo[d]thiazole;
5-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)benzo[d]thiazole;
5-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butoxy)-lH-benzo[d]imidazol-2(3H)-one;
7- (4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)-3,4-dihydroisoquinolin-l(2H)-one; 7-((4-((4-(2,3-dichlorophenyl)piperazin-l-yl)methyl)cyclohexyl)methoxy)-3,4-dihydroquinoli 2(lH)-one;
7-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)but-2-ynyloxy)-3,4-dihydroquinolin-2(lH)-one;
6- (4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)-lH-indazole;
(E)-7-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)but-2-enyloxy)-3,4-dihydroquinolin-2(lH)-one;
7- (3-((4-(2,3-dichlorophenyl)piperazin-l-yl)methyl)phenoxy)-3,4-dihydroquinolin-2(lH)-one; 7-(4-((4-(2,3-dichlorophenyl)piperazin-l-yl)methyl)phenoxy)-3,4-dihydroquinolin-2(lH)-one; 7-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propoxy)-3,4-dihydroisoquinolin-l(2H)-one;
7-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butoxy)-3,4-dihydroisoquinolin-l(2H)-one;
5 -(3 -(4-(2,3 -dichlorophenyl)piperazin- 1 -yl)propoxy)- 1 H-benzo [d] imidazol-2(3H)-one;
5-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)-lH-benzo[d]imidazol-2(3H)-one;
5- (4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)benzo[d]thiazole;
7- (4-(4-(2-methoxyphenyl)piperazin-l-yl)butoxy)-3,4-dihydro-l,8-naphthyridin-2(lH)-one;
8- (4-(4-(2-oxo-l,2,3,4-tetrahydroquinolin-7-yloxy)butyl)piperazin-l-yl)-2H-benzo[b][l,4]oxazin- 3(4H)-one;
7-(4-(4-(2-methoxyphenyl)piperazin-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)-one;
7-(4-(4-(2,3-dihydrobenzo[b][l,4]dioxin-5-yl)piperazin-l-yl)butoxy)-3,4-dihydroquinolin-2(lH)- one;
7-(4-(4-(2,3-dihydrobenzo[b][l,4]dioxin-5-yl)piperazin-l-yl)butoxy)-3,4-dihydro-l,8- naphthyridin-2(lH)-one;
7-(4-(4-(2,3-dihydrobenzo[b][l,4]dioxin-5-yl)piperazin-l-yl)butoxy)quinolin-2(lH)-one;
N-(5-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butoxy)-2-methylphenyl)acetamide;
6- (3-(4-(2,3-dichlorophenyl)piperidin-l-yl)propoxy)-lH-indazole;
6- (4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butoxy)-lH-indazole;
7- (4-(3-(lH-indazol-6-yloxy)propyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one;
7-(4-(4-(lH-indazol-6-yloxy)butyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one;
6-(3-(4-(2-methoxyphenyl)piperazin-l-yl)propoxy)-lH-indazole; and
6-(4-(4-(2-methoxyphenyl)piperazin-l-yl)butoxy)-lH-indazole.
23. The compound of claim 14, selected from the group consisting of:
N-(3-(4-(2,3-dichlorophenyl)piperidin-l -yl)propyl)- lH-indazol-6-amine;
N-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butyl)-lH-indazol-6-amine;
N-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propyl)-lH-indazol-6-amine;
N-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butyl)-lH-indazol-6-amine;
N-(3-(4-(2-methoxyphenyl)piperazin-l-yl)propyl)-lH-indazol-6-amine; N-(4-(4-(2-methoxyphenyl)piperazin- 1 -yl)butyl)- lH-indazol-6-amine;
7-(4-(3 -( 1 H-indazol-6-ylamino)propyl)piperazin- 1 -yl)benzo [d]oxazol-2(3H)-one;
7-(4-(4-(lH-indazol-6-ylamino)butyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one;
N-(3-(4-(2,3-dichlorophenyl)piperidin-l-yl)propyl)benzo[d]thiazol-5-amine;
N-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butyl)benzo[d]thiazol-5-amine;
N-(3-(4-(2,3-dichlorophenyl)piperazin-l-yl)propyl)benzo[d]thiazol-5-amine;
N-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butyl)benzo[d]thiazol-5-amine;
5 -(3 -(4-(2-methoxyphenyl)piperazin- 1 -yl)propoxy)benzo [d]thiazole;
5-(4-(4-(2-methoxyphenyl)piperazin-l-yl)butoxy)benzo[d]thiazole;
N-(3-(4-(2-methoxyphenyl)piperazin-l-yl)propyl)benzo[d]thiazol-5-amine;
N-(4-(4-(2-methoxyphenyl)piperazin-l-yl)butyl)benzo[d]thiazol-5-amine;
7-(4-(3-(benzo[d]thiazol-5-yloxy)propyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one;
7-(4-(4-(benzo[d]thiazol-5-yloxy)butyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one;
7-(4-(3-(benzo[d]thiazol-5-ylamino)propyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one;
7-(4-(4-(benzo[d]thiazol-5-ylamino)butyl)piperazin-l-yl)benzo[d]oxazol-2(3H)-one;
5 -(3 -(4-(2,3 -dichlorophenyl)piperidin- 1 -yl)propylamino)- 1 H-benzo [d] imidazol-2(3H)-one;
5-(4-(4-(2,3-dichlorophenyl)piperidin-l-yl)butylamino)-lH-benzo[d]imidazol-2(3H)-one;
5 -(3 -(4-(2,3 -dichlorophenyl)piperazin- 1 -yl)propylamino)- 1 H-benzo [d]imidazol-2(3H)-one;
5-(4-(4-(2,3-dichlorophenyl)piperazin-l-yl)butylamino)-lH-benzo[d]imidazol-2(3H)-one;
5-(3-(4-(2-methoxyphenyl)piperazin-l-yl)propoxy)-lH-benzo[d]imidazol-2(3H)-one;
5-(4-(4-(2-methoxyphenyl)piperazin-l-yl)butoxy)-lH-benzo[d]imidazol-2(3H)-one;
5-(3-(4-(2-methoxyphenyl)piperazin-l-yl)propylamino)-lH-benzo[d]imidazol-2(3H)-one;
5-(4-(4-(2-methoxyphenyl)piperazin-l-yl)butylamino)-lH-benzo[d]imidazol-2(3H)-one;
7-(4-(3-(2-oxo-2,3-dihydro-lH-benzo[d]imidazol-5-yloxy)propyl)piperazin-l-yl)benzo[d]oxazol-
2(3H)-one;
7-(4-(4-(2-oxo-2,3-dihydro-lH-benzo[d]imidazol-5-yloxy)butyl)piperazin-l-yl)benzo[d]oxazol- 2(3H)-one;
7-(4-(3-(2-oxo-2,3-dihydro-lH-benzo[d]imidazol-5-ylamino)propyl)piperazin-l- yl)benzo[d]oxazol-2(3H)-one; and
7-(4-(4-(2-oxo-2,3-dihydro-lH-benzo[d]imidazol-5-ylamino)butyl)piperazin-l- yl)benzo [d] oxazol-2(3H)-one.
24. A pharmaceutical composition comprising the compound of any one of claims 1- 23 and a pharmaceutically acceptable carrier.
25. A method of modulating the activity of a D2 dopamine receptor comprising contacting the receptor with a compound of any one of claims 1-23.
26. A method of treating a central nervous system disorder associated with D2 dopamine receptors in a subject, comprising delivering to the subject a therapeutically effective amount of the compound of any one of claims 1-23.
27. The method of claim 26, wherein the disorder associated with D2 dopamine receptors is a psychiatric disorder.
28. The method of claim 27, wherein the psychiatric disorder is selection from the group consisting of schizophrenia, schizoaffective disorder, schizophreniform disorder, bipolar disorder, mania, manic psychosis, Tourette's syndrome and Tourette-like disorders, obsessive- compulsive disorder, depression, bipolar depression, unipolar depression, depression with psychotic features, psychosis not otherwise specified, anxiety disorders, affective disorders, and non-affective disorders.
29. The method of claim 26, wherein the disorder associated with D2 dopamine receptors is a neurological disorder.
30. The method of claim 29, wherein the neurological disorder is selected from the group consisting of Huntington's Disease, Parkinsonian Psychosis, autism spectrum and autistic disorders, Alzheimer's Disease, dementia, and iemann-Pick Disorder.
31. The method of claim 26, wherein the disorder associated with D2 dopamine receptors is a pituitary disorder.
32. The method of claim 31, wherein the pituitary disorder is selected from the group consisting of pituitary adenoma and prolactinoma.
33. The method of claim 26, wherein the disorder associated with D2 dopamine receptors is an endocrine disorder.
34. The method of claim 33, wherein the endocrine disorder is galactorrhea.
35. The method of claim 26, further comprising delivering to the subject an additional therapeutic agent.
36. The method of claim 35, wherein the compound is administered in the same pharmaceutical composition as the additional therapeutic agent.
37. The method of claim 35, wherein the compound is administered in a different pharmaceutical composition than the additional therapeutic agent.
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