WO2008060568A2 - Amide-substituted aryl piperidines - Google Patents

Amide-substituted aryl piperidines Download PDF

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WO2008060568A2
WO2008060568A2 PCT/US2007/023936 US2007023936W WO2008060568A2 WO 2008060568 A2 WO2008060568 A2 WO 2008060568A2 US 2007023936 W US2007023936 W US 2007023936W WO 2008060568 A2 WO2008060568 A2 WO 2008060568A2
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alkyl
compound
amino
hydrate
salt
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PCT/US2007/023936
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French (fr)
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WO2008060568A3 (en
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Rajagopal Bakthavatchalam
Scott M. Capitosti
Alan J. Hutchison
Guiying Li
John M. Peterson
Charles S. Cheng
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Neurogen Corporation
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Publication of WO2008060568A3 publication Critical patent/WO2008060568A3/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—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 three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D519/00—Heterocyclic compounds containing more than one system of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring system not provided for in groups C07D453/00 or C07D455/00

Definitions

  • This invention relates generally to amide-substituted aryl piperidines and related compounds, and to the use of such compounds to treat conditions responsive to calcitonin gene-related peptide (CGRP) receptor modulation.
  • the invention further relates to the use of such compounds as reagents for the identification of other agents that bind to CGRP receptor, and as probes for the detection and localization of CGRP receptor.
  • Calcitonin gene-related peptide is a naturally occurring peptide that is widely distributed in both the peripheral and central nervous systems. This peptide is primarily localized in sensory afferent and central neurons, where it exerts a number of biological effects, including vasodilation. CGRP is released from trigeminal ganglia nerves after nerve activation and is a strong cerebral and dural vessel dilator. Accordingly, CGRP regulates blood flow to the brain and meninges (which are pain- sensitive), and is involved in the pathophysiology of conditions such as migraine and cluster headaches.
  • CGRP Calcitonin gene-related peptide
  • CGRP exerts its biological actions by binding to specific cell surface receptors (CGRP receptors), which are G-protein coupled and in turn activate intracellular adenylate cyclase.
  • CGRP receptors generally have three discrete components: (1) a transmembrane calcitonin receptor-like receptor (CRLR or CLR); (2) a transmembrane receptor activity modifying protein type one (RAMPl) and (3) an intracellular receptor component protein (RCP), all of which are required in order to form a functional CGRP receptor complex.
  • RAMPl transmembrane receptor activity modifying protein type one
  • RCP intracellular receptor component protein
  • the present invention provides amide-substituted aryl piperidines that satisfy Formula A, Formula I or Formula II:
  • ⁇ represents a single or double bond; are each a 5- or 6-membered heterocycle that:
  • (a) comprises at least one ring nitrogen atom
  • (c) is optionally substituted with Ci-C 4 alkyl
  • Ar is a 5- or 6-membered heteroaryl, each of which is optionally substituted and each of which is preferably substituted with from 0 to 2 substituents independently chosen from R 3 ;
  • Z is C, CH, CH 2 , N or NH;
  • n is 0, 1 or 2;
  • R] is C]-C 8 aminoalkyl, Ci-C 8 hydroxyalkyl, C 2 -C 8 alkyl ether, phenylC 0 -C 6 alkyl, or (4- to 12-membered heterocycle)C 0 -C 6 alkyl, each of which is optionally substituted and each of which is preferably substituted with from 0 to 6 substituents independently chosen from:
  • R 2 is hydrogen, C]-C 6 alkyl or Ci-C 6 haloalkyl; or Ri and R 2 are taken together to form a 4- to 12-membered heterocycle that is optionally substituted and is preferably substituted with from 0 to 6 substituents independently chosen from:
  • Ci-C 6 alkyl C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, C]-C 6 haloalkyl, Ci-C 6 hydroxyalkyl, C 2 -C 6 alkyl ether, Ci-C 6 alkoxy, Ci-C 6 alkoxycarbonyl, (C 3 -C 8 cycloalkyl)C 0 -C 4 alkyl, mono- or di- (Ci-C 6 alkyl)aminoC 0 -C 4 alkyl, phenylC 0 -C 4 alkyl, (4- to 12-membered heterocycle)Co-C 4 alkyl, and spiro C 3 -C 8 cycloalkyl; each of which is optionally substituted and each of which is preferably substituted with from 0 to 6 substituents independently
  • Each R 3 is independently chosen from:
  • Ci-C 6 alkyl (i) halogen, hydroxy, cyano and amino; and (ii) Ci-C 6 alkyl, (C 3 -C 8 carbocycle)C 0 -C 4 alkyl, Ci-C 6 haloalkyl, Ci-C 6 alkoxy, Ci-
  • C 6 alkyl ether mono- or di-(Ci-C 6 alkyl)aminoC 0 -C 2 alkyl, and (4- to 7-membered heterocycle)C 0 -C 4 alkyl; each of which is optionally substituted and each of which is preferably substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, Ci-C 4 alkyl, (C 3 -C 8 carbocycle)C 0 -C 4 alkyl and (4- to 10-membered heterocycle)Co-C 4 alkyl; or two substituents represented by R 3 are taken together to form a fused 5- to 7- membered carbocycle or heterocycle that is optionally substituted and is preferably substituted with from 0 to 3 substituents independently chosen from halogen, hydroxy, amino, Ci-C 6 alkyl, (C 3 -C 8 carbocycle)C 0 -C 4 alkyl, Ci-C ⁇ haloalkyl, Ci-C
  • R 4 represents:
  • Ci-C 6 alkyl C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, Ci-C 6 haloalkyl, Q-Cehydroxyalkyl, Ci-C 6 alkoxy, Ci-C 6 alkoxycarbonyl, (C 3 -C 8 cycloalkyl)Co-C 4 alkyl, mono- or di- (C 1 -
  • R 5 represents from 0 to 4 substituents independently chosen from Ci-C 6 alkyl, C 2 - C 6 alkenyl, C 2 -C 6 alkynyl and (C 3 -C 8 cycloalkyl)Co-C 4 alkyl; and R 6 represents from O to 4 substituents independently chosen from halogen, hydroxy, amino, cyano, Ci-C 6 alkyl, Ci-C 6 haloalkyl, Ci-C 6 alkoxy and Ci-C 6 alkyl ether.
  • the present invention provides amide-substituted aryl piperidines that satisfy Formula III or IV:
  • Z is C, CH, CH 2 , N or NH; ⁇ represents a single or double bond; n is 0, 1 or 2;
  • A, B, D and E are each C, CH or N, such that exactly one or two of A, B, D and E are N;
  • T, U and V are independently C, CH, N, NH, S and O, such that at least one of T,
  • U and V is N; exactly one of T, U and V is CH; and aromatic; and R 3 represents from 0 to 3 substituents independently chosen from:
  • the present invention provides amide-substituted aryl piperidines that satisfy Formula V or VI:
  • A, B, D and E are each C, CH or N, such that exactly one or two of A, B, D and E are N;
  • T, U and V are independently C, CH, N, NH, S and O, such that at least one of T,
  • U and V is N; exactly one of T, U and V is CH; and
  • R 3 represents from 0 to 3 substituents independently chosen from:
  • X is C
  • such amide-substituted aryl piperidines further satisfy Formula VIII or IX:
  • X is C
  • Z is C, CH, CH 2 , N or NH
  • n 0, 1 or 2; and the remaining variables are as described above.
  • amide-substituted aryl piperidines provided herein are CGRP receptor modulators and exhibit an EC 50 or IC 50 of no greater than 10 micromolar, 5 micromolar, 2 micromolar, 1 micromolar, 500 nanomolar, 100 nanomolar, 50 nanomolar or 10 nanomolar in an assay for determination of CGRP receptor agonist or antagonist activity.
  • CGRP receptor modulators provided herein are CGRP receptor antagonists; in certain embodiments, such antagonists exhibit no detectable CGRP receptor agonist activity above background at their EC 50 for CGRP receptor.
  • amide-substituted aryl piperidines provided herein are labeled with a detectable marker (e.g., radiolabeled or fluorescein conjugated).
  • a detectable marker e.g., radiolabeled or fluorescein conjugated
  • the present invention further provides pharmaceutical compositions comprising at least one amide-substituted aryl piperidine provided herein in combination with a physiologically acceptable carrier or excipient.
  • Methods are further provided for inhibiting agonist-induced CGRP receptor activity.
  • the inhibition takes place in vitro.
  • Such methods comprise contacting a CGRP receptor with at least one CGRP receptor antagonist as described herein, under conditions and in an amount or concentration sufficient to detectably inhibit agonist-induced CGRP receptor activity.
  • the CGRP receptor is in a patient.
  • Such methods comprise contacting cells expressing a CGRP receptor in a patient with a CGRP receptor antagonist as described herein in an amount or concentration that would be sufficient to detectably inhibit agonist-induced CGRP receptor activity in cells expressing CGRP receptor in vitro.
  • the present invention further provides methods for treating a condition responsive to CGRP receptor modulation in a patient, comprising administering to the patient a therapeutically effective amount of at least one amide-substituted aryl piperidine provided herein.
  • methods for treating migraine or other headache in a patient, comprising administering to a patient suffering from (or at risk for) headache a therapeutically effective amount of at least one amide-substituted aryl piperidine provided herein.
  • the present invention provides methods for determining the presence or absence of CGRP receptor in a sample, comprising: (a) contacting a sample with an amide-substituted aryl piperidine provided herein under conditions that permit binding of the compound to CGRP receptor; and (b) detecting a signal indicative of a level of the compound bound to CGRP receptor.
  • the present invention provides methods of preparing the compounds disclosed herein, including the intermediates.
  • methods for detecting CGRP receptor antagonist activity of a test compound comprising: (i) contacting test cells that stably expresses CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2 with: (a) a test compound; (b) an indicator of intracellular or cytosolic calcium; and (c) CGRP receptor agonist; (ii) detecting a response indicative of intracellular or cytosolic calcium level in the contacted test cells; (iii) contacting matched control cells that stably expresses CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2 with: (a) an indicator of intracellular or cytosolic calcium; and (b) CGRP receptor agonist; (iv) detecting a response indicative of intracellular or cytosolic calcium level in the contacted control
  • the present invention provides amide-substituted aryl piperidines, which may be used in vitro or in vivo in a variety of contexts, as described herein.
  • amide-substituted aryl piperidine encompasses any compound that satisfies Formula A or Formula I or Formula II or Formula VII, and optionally further satisfies one or more additional formulas provided herein. This term further includes pharmaceutically acceptable salts, solvates ⁇ e.g., hydrates) and esters of such compounds.
  • a "pharmaceutically acceptable salt” of a compound recited herein is an acid or base salt that is suitable for use in contact with the tissues of human beings or animals without excessive toxicity or carcinogenicity, and preferably without irritation, allergic response, or other problem or complication.
  • Specific pharmaceutically acceptable anions for use in salt formation include, but are not limited to, acetate, 2-acetoxybenzoate, ascorbate, benzoate, bicarbonate, bromide, calcium edetate, carbonate, chloride, citrate, dihydrochloride, diphosphate, ditartrate, edetate, estolate (ethylsuccinate), formate, fumarate, gluceptate, gluconate, glutamate, glycolate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroiodide, hydroxymaleate, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, pamoate, pantothenate, phenylacetate
  • pharmaceutically acceptable cations for use in salt formation include, but are not limited to ammonium, benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, and metals such as aluminum, calcium, lithium, magnesium, potassium, sodium and zinc.
  • a pharmaceutically acceptable acid or base salt can be synthesized from a parent compound that contains a basic or acidic moiety by any conventional chemical method.
  • such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, the use of nonaqueous media, such as ether, ethyl acetate, ethanol, methanol, isopropanol or acetonitrile, is preferred.
  • nonaqueous media such as ether, ethyl acetate, ethanol, methanol, isopropanol or acetonitrile
  • prodrugs of the compounds provided herein are a compound that may not fully satisfy the structural requirements of a formula provided herein, but is modified in vivo, following administration to a patient, to produce a compound within the scope of such formula.
  • a prodrug may be an acylated derivative of a compound as provided herein.
  • Prodrugs include compounds wherein hydroxy, amine or sulfhydryl groups are bonded to any group that, when administered to a mammalian subject, cleaves to form a free hydroxy, amino or sulfhydryl group, respectively.
  • Examples of prodrugs include, but are not limited to, acetate, formate, phosphate and benzoate derivatives of alcohol and amine functional groups within the compounds provided herein.
  • Prodrugs of the compounds provided herein may be prepared by modifying functional groups present in the compounds in such a way that the modifications are cleaved in vivo to yield the parent compounds.
  • alkyl refers to a straight or branched chain saturated aliphatic hydrocarbon.
  • Alkyl groups include groups having from 1 to 8 carbon atoms (Cj- C 8 alkyl), from 1 to 6 carbon atoms (Ci-C 6 alkyl) and from 1 to 4 carbon atoms (Ci-
  • C 4 alkyl such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2- pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl or 3-methylpentyl.
  • a substituent of an alkyl group is specifically indicated.
  • Ci- C 6 aminoalkyl refers to a Ci-C 6 alkyl that has at least one -NH 2 substituent
  • Ci- Qhydroxyalkyl refers to a Ci -Qalkyl substituted with at least one -OH.
  • C 0 -C 4 alkyl refers to a single covalent bond (C 0 ) or an alkylene group having 1, 2, 3 or 4 carbon atoms; "C 0 -C 2 alkyl” refers to a single covalent bond or a methylene or ethylene group.
  • Alkylene refers to a divalent alkyl group, as defined above. Ci-C 4 alkylene is an alkylene group having 1, 2, 3 or 4 carbon atoms.
  • Alkenyl refers to straight or branched chain alkene groups, which comprise at least one unsaturated carbon-carbon double bond. Alkenyl groups include C 2 -C 8 alkenyl, C 2 -C 6 alkenyl and C 2 -C 4 alkenyl groups, which have from 2 to 8, 2 to 6 or 2 to 4 carbon atoms, respectively, such as ethenyl, allyl or isopropenyl.
  • Alkynyl refers to straight or branched chain alkyne groups, which have one or more unsaturated carbon-carbon bonds, at least one of which is a triple bond.
  • Alkynyl groups include C 2 -C 8 alkynyl, C 2 -C 6 alkynyl and C 2 -C 4 alkynyl groups, which have from 2 to 8, 2 to 6 or 2 to 4 carbon atoms, respectively.
  • a “cycloalkyl” is a saturated or partially saturated cyclic group in which all ring members are carbon, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and partially saturated variants thereof. Certain cycloalkyl groups are C 3 -C 8 cycloalkyl, in which the ring contains from 3 to 8 ring members, all of which are carbon.
  • a "(C 3 - C 8 cycloalkyl)C 0 -C 4 alkyl” is a C 3 -C 8 cycloalkyl group linked via a single covalent bond or a Ci-C 4 alkylene group.
  • a "spiro C 3 -C 8 cycloalkyl” is a C 3 -C 8 cycloalkyl substituent of a ring attached so as to form a spiro ring system. For example, if R 7 in the group designated: is a spiro cyclopropyl ring, one resulting ring system is
  • alkoxy is meant an alkyl group attached via an oxygen bridge ⁇ i.e., -O-alkyl).
  • Alkoxy groups include Ci-C 6 alkoxy and Ci-C 4 alkoxy groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively.
  • Methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy and 3-methylpentoxy are representative alkoxy groups.
  • alkylthio refers to an alkyl group as attached via a sulfur bridge ⁇ i.e., - S-alkyl).
  • Alkylthio groups include Ci-C 6 alkylthio and Ci-C 4 alkylthio groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively.
  • Alkylsulfinyl refers to groups of the formula -(SO)-alkyl, in which the sulfur atom is the point of attachment.
  • Alkylsulfinyl groups include Ci-C 6 alkylsulfinyl and Ci- C 4 alkylsulfmyl groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively.
  • Alkylsulfonyl refers to groups of the formula -(SO 2 )-alkyl, in which the sulfur atom is the point of attachment.
  • Alkylsulfonyl groups include Ci-C 6 alkylsulfonyl and Ci- C 4 alkylsulfonyl groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively.
  • the term "(Ci-C 6 alkylsulfonyl)C 0 -C 4 alkyl refers to a Ci-C 6 alkylsulfonyl that is linked via a single covalent bond or a Ci-C 4 alkylene group ⁇ i.e., -(C 0 -C 4 alkyl)-(SO 2 )-(Ci-C 6 alkyl)).
  • alkoxycarbonyl refers to an alkoxy group linked via a carbonyl ⁇ i.e., a group having the general structure -C ⁇ CO-O-alkyl).
  • Alkoxycarbonyl groups include Ci- C 8 , Ci-C 6 and Ci-C 4 alkoxycarbonyl groups, which have from 1 to 8, 6 or 4 carbon atoms, respectively, in the alkyl portion of the group.
  • Alkanoyl groups have the indicated number of carbon atoms, with the carbon of the keto group being included in the numbered carbon atoms.
  • Alkanoyl groups include, for example, C]-C 8 alkanoyl, Ci-C 6 alkanoyl and Ci-C 4 alkanoyl groups, which have from 1 to 8, from 1 to 6 or from 1 to 4 carbon atoms, respectively.
  • Alkanoylamino groups include, for example, (Ci- C 6 alkanoyl)amino groups, which have from 1 to 6 carbon atoms within the alkanoyl group.
  • Alkylamino refers to a secondary or tertiary amine that has the general structure -NH-alkyl or -N(alkyl)(alkyl), wherein each alkyl is selected independently from alkyl, cycloalkyl and (cycloalkyl)alkyl groups.
  • groups include, for example, mono- and di- (Ci-C 8 alkyl)amino groups, in which each Ci-C 8 alkyl may be the same or different, as well as mono- and di-(Ci-C 6 alkyl)amino groups and mono- and di-(Ci-C 4 alkyl)amino groups.
  • Alkylaminoalkyl refers to an alkylamino group linked via an alkylene moiety (i.e., a group having the general structure -alkylene-NH-alkyl or -alkylene-
  • Alkylaminoalkyl groups include, for example, mono- and di- (Ci-C 8 alkyl)aminoCi-C 6 alkyl, and mono- and di-(Ci-C 6 alkyl)aminoCi-C 4 alkyl.
  • "Mono- or di-(C]-C 6 alkyl)aminoC 0 -C 4 alkyl” refers to a mono- or di-(Ci-C ⁇ alkyl)amino group linked via a single covalent bond or a Ci-C 4 alkylene group.
  • alkyl as used in the terms “alkylamino” and “alkylaminoalkyl” differs from the definition of "alkyl” used for all other alkyl- containing groups, in the inclusion of cycloalkyl and (cycloalkyl)alkyl groups (e.g., (C 3 - C 7 cycloalkyl)C 0 -C 2 alkyl).
  • aminonosulfonyl refers to a sulfonamide group (i.e., -SO 2 NH 2 ).
  • di-(Ci-C 6 alkyl)aminosulfonyl refers to groups of the formula -(SO 2 )- N(R) 2 , in which the sulfonyl is the point of attachment, one R is C]-C 6 alkyl and the other R is hydrogen or an independently chosen Ci-C 6 alkyl.
  • halogen refers to fluorine, chlorine, bromine or iodine.
  • haloalkyl is an alkyl group that is substituted with 1 or more independently chosen halogens (e.g., "Ci-Cshaloalkyl” groups have from 1 to 8 carbon atoms; "C]- C 6 haloalkyl” groups have from 1 to 6 carbon atoms).
  • haloalkyl groups include, but are not limited to, mono-, di- or tri-fluoromethyl; mono-, di- or tri- chloromethyl; mono-, di-, tri-, tetra- or penta-fluoroethyl; mono-, di-, tri-, tetra- or penta- chloroethyl; and 1,2,2,2-tetrafluoro-l-trifluoromethyl-ethyl.
  • Typical haloalkyl groups are trifiuoromethyl and difluoromethyl.
  • a dash (“-") that is not between two letters or numbers is used to indicate a point of attachment for a substituent.
  • a “carbocycle” or “carbocyclic group” comprises at least one ring formed entirely by carbon-carbon bonds (referred to herein as a carbocyclic ring), and does not contain a heterocycle. Unless otherwise specified, each ring within a carbocycle may be independently saturated, partially saturated or aromatic, and is optionally substituted as indicated.
  • a carbocycle generally has from 1 to 3 fused, pendant or spiro rings; carbocycles within certain embodiments have one ring or two fused rings. Typically, each ring contains from 3 to 8 ring members (i.e., C 3 -C 8 ); rings having from 5 to 7 ring members are recited in certain embodiments.
  • Carbocycles comprising fused, pendant or spiro rings typically contain from 9 to 14 ring members. Certain representative carbocycles are cycloalkyl as described above. Other carbocycles are aryl (i.e., contain at least one aromatic carbocyclic ring, with or without one or more additional aromatic and/or cycloalkyl rings). Such aryl carbocycles include, for example, phenyl, naphthyl (e.g., 1 -naphthyl and 2-naphthyl), fluorenyl, indanyl and 1,2,3,4-tetrahydronaphthyl.
  • Certain 3- to 8-membered carbocycles are linked via a single covalent bond or Ci- C 4 alkylene; and are accordingly designated "(C 3 -C 8 carbocycle)C 0 -C 4 alkyl.”
  • Such groups include, for example, aryl and arylalkyl moieties (such as phenyl, indanyl, and groups in which either of the foregoing is linked via Ci-C 4 alkylene; for example, a phenylCo-C 4 alkyl refers to a phenyl group linked via a single covalent bond or Ci-C 4 alkylene) and (cycloalkyl)alkyl moieties as described above.
  • a “heterocycle” (also referred to herein as a “heterocyclic group”) has from 1 to 3 fused, pendant or spiro rings, at least one of which is a heterocyclic ring (i.e., one or more ring atoms is a heteroatom independently chosen from oxygen, sulfur and nitrogen, with the remaining ring atoms being carbon).
  • a heterocyclic ring comprises 1, 2, 3 or 4 heteroatoms; within certain embodiments each heterocyclic ring has 1 or 2 heteroatoms per ring.
  • Each heterocyclic ring generally contains from 4 to 8 ring members (rings having from 4 or 5 to 7 ring members are recited in certain embodiments) and heterocycles comprising fused, pendant or spiro rings typically contain from 9 to 14 ring members.
  • Certain heterocycles comprise a sulfur atom as a ring member; in certain embodiments, the sulfur atom is oxidized to SO or SO 2 .
  • Heterocycles may be optionally substituted with a variety of substituents, as indicated.
  • Certain heterocycles are 4- to 10- membered or 4- to 10-membered and comprise one or two rings; in certain embodiments, such heterocycles are monocyclic (e.g., 4- to 8-membered, 5- to 8-membered, 4- to 7- membered, or 5- or 6-membered).
  • heterocycles are heteroaryl groups (i.e., at least one heterocyclic ring within the group is aromatic), such as a 5- or 6-membered heteroaryl (e.g., thienyl, imidazolyl, pyridyl or pyrimidyl).
  • heterocycles are heterocycloalkyl groups (i.e., do not comprise an aromatic heterocyclic ring).
  • a heterocycle may be linked by a single covalent bond or an alkylene group; for example, "(4- to 12-membered heterocycle)Co-C 4 alkyl” refers to a 4- to 12-membered heterocycle having 1 or 2 rings, which is linked via a single covalent bond or via an alkylene group having from 1 to 6 carbons. "(4- to 7-membered heterocycle)Co-C 4 alkyl” refers to a 4- to 7-membered heterocyclic ring that is linked via a single covalent bond or an alkylene group having from 1 to 4 carbons, methylene or ethylene linker. When substituted, it will be apparent that substituent(s) may be attached on the ring and/or the alkylene linker.
  • a "substituent,” as used herein, refers to a molecular moiety that is covalently bonded to an atom within a molecule of interest.
  • a "ring substituent” may be a moiety such as a halogen, alkyl group, haloalkyl group or other group discussed herein that is covalently bonded to an atom (such as a carbon or nitrogen atom) that is a ring member.
  • substitution refers to replacing a hydrogen atom in a molecular structure with a substituent as described above, such that the valence on the designated atom is not exceeded, and such that a chemically stable compound (i.e., a compound that can be isolated, characterized, and tested for biological activity) results from the substitution.
  • Groups that are "optionally substituted” are unsubstituted or are substituted by other than hydrogen at one or more available positions, typically 1, 2, 3, 4 or 5 positions, by one or more suitable groups (which may be the same or different).
  • Optional substitution is also indicated by the phrase "substituted with from 0 to X substituents," where X is the maximum number of possible substituents.
  • Certain optionally substituted groups are substituted with from 0 to 2, 3 or 4 independently selected substituents (i.e., are unsubstituted or substituted with up to the recited maximum number of substituents).
  • CGRP receptor refers to any functional CGRP receptor complex, comprising a ligand binding protein CRLR and a chaperone protein RAMPl .
  • a receptor complex further comprises a coupling protein RCP.
  • each complex component is human: human CRLR includes, for example, the polypeptide having the sequence provided at Genbank accession number NM_005795, and human RAMPl includes, for example, the polypeptide having the sequence provided at Genbank accession number NM 005855.
  • One or more components of the receptor complex may be expressed endogenously; in certain embodiments it is convenient to use cells that express CRLR and RAMPl recombinantly, with RCP expressed endogenously.
  • CGRP receptor agonist refers to a compound that binds CGRP receptor and induces signal transduction mediated by CGRP receptor.
  • CGRP receptor agonists include, for example, CGRP (e.g., human CGRP), as well as peptide portions or variants thereof that bind CGRP receptor and retain CGRP-receptor activating activity.
  • a “CGRP receptor antagonist” is a compound that detectably inhibits signal transduction mediated by CGRP receptor. Such inhibition may be determined using the representative calcium mobilization assay provided in Example 5.
  • Preferred CGRP receptor antagonists have an IC 50 of 10 ⁇ M or less in this assay, more preferably 5 ⁇ M or less, and still more preferably 1 ⁇ M or less, 500 nM or less, or 100 nM or less.
  • Certain CGRP receptor antagonists exhibit minimal agonist activity (i.e., they induce an increase in the basal activity of CGRP receptor that is less than 5% of the increase that would be induced by one EC 50 of the peptide agonist CGRP) and more preferably exhibit no detectable agonist activity within the assay described in Example 5.
  • CGRP receptor antagonists for use as described herein are generally non-toxic.
  • CGRP receptor antagonists include neutral antagonists and inverse agonists.
  • a "neutral antagonist" of CGRP receptor is a compound that inhibits the activity of
  • CGRP receptor agonist at CGRP receptor but when present at its IC 50 (determined using the calcium mobilization assay as described in Example 5 does not significantly change the basal activity of the receptor.
  • the effect on the basal activity may be determined using the calcium mobilization assay described in Example 5 performed in the absence of agonist.
  • CGRP receptor activity is reduced by no more than 10%, more preferably by no more than 5%, and even more preferably by no more than 2%; most preferably, there is no detectable reduction in activity.
  • Neutral antagonists may, but need not, also inhibit the binding of agonist to CGRP receptor.
  • An "inverse agonist" of CGRP receptor is a compound that reduces the activity of CGRP receptor below its basal activity level in the absence of activating concentrations of agonist. Inverse agonists may also inhibit the activity of agonist at CGRP receptor, and/or may inhibit binding of CGRP receptor agonist to CGRP receptor. The reduction in basal activity of CGRP receptor produced by an inverse agonist may be determined from a calcium mobilization assay, such as the assay of Example 5.
  • a "therapeutically effective amount" (or dose) is an amount that, upon administration to a patient, results in a discernible patient benefit (e.g., provides detectable relief from a condition being treated). Such relief may be detected using any appropriate criteria.
  • a therapeutically effective amount or dose generally results in a concentration of compound(s) in a body fluid (such as blood, plasma, serum, CSF, synovial fluid, lymph, cellular interstitial fluid, tears or urine) that is sufficient to result in detectable alteration in CGRP receptor -mediated signal transduction (using an assay provided herein).
  • a body fluid such as blood, plasma, serum, CSF, synovial fluid, lymph, cellular interstitial fluid, tears or urine
  • the discernible patient benefit may be apparent after administration of a single dose, or may become apparent following repeated administration of the therapeutically effective dose according to a predetermined regimen, depending upon the indication for which the compound is administered.
  • a “patient” is any individual treated with an amide-substituted aryl piperidine as provided herein.
  • Patients include humans, as well as other animals such as companion animals (e.g., dogs and cats) and livestock; in certain embodiments, human patients are preferred.
  • Patients may be experiencing one or more symptoms of a condition responsive to CGRP receptor modulation or may be free of such symptom(s) (i.e., treatment may be prophylactic in a patient considered to be at risk for the development of such symptoms).
  • AMIDE-SUBSTITUTED ARYL PIPERIDINES As noted above, the present invention provides amide-substituted aryl piperidines that satisfy any one of Formulas A, I, II, or VII, above. Such amide-substituted aryl piperidines may be used in a variety of contexts, including in the treatment of conditions responsive to CGRP receptor modulation. Such compounds may also be used within in vitro assays (e.g., assays for CGRP receptor activity), as probes for the detection and localization of CGRP receptor and within assays to identify other CGRP receptor antagonists.
  • in vitro assays e.g., assays for CGRP receptor activity
  • Certain amide-substituted aryl piperidines of Formula A or Formula I or Formula II further satisfy one of Formulas III- VI, as described above.
  • Certain amide-substituted aryl piperidines of Formula VII further satisfy Formula VIII of Formula IX, as described above.
  • R 7 represents from 0 to 4 substituents independently chosen from: (i) halogen, hydroxy, amino and oxo; and (ii) Ci-C ⁇ alkyl, Ci-C 6 alkoxy, mono- or di-(Ci-C 6 alkyl)amino, Ci- C 6 alkoxycarbonyl, C 2 -C 6 alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C 3 - C 7 cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C 4 alkyl, Ci-C 4 alkoxy and mono- or di-(Cp C 4 alkyl) amino; and are each a 5- or 6-membered heterocycle (e.g., a 5-membered heteroaryl) that is optionally substituted with Ci-C 4 alkyl.
  • substituents independently chosen from: (i) halogen, hydroxy
  • R 7 moieties in the above formulas include methyl (e.g., in certain compounds, R 7 represents two methyl substituents, such as gem-dimethyl) and spiro C 3 - C 6 cycloalkyl groups. Within certain such compounds, the group designated:
  • each R 9 is independently hydrogen or Ci-
  • Ci-C 4 alkyl include: an( j m which m is an integer ranging from 1 to 4.
  • R 7 moieties within such compounds include, for example hydroxy, amino, oxo, Ci-C ⁇ alkyl, Ci-C 6 hydroxyalkyl, C]-C 6 alkoxy, mono- or di-(Ci-C 6 alkyl)amino, (Ci-C 6 alkanoyl)amino, Ci-C 6 alkoxycarbonyl, C 2 -C 6 alkyl ether, phenyl and 4- to 7- membered heterocycles.
  • o is 1 (i.e.,
  • R 7 represents two methyl substituents.
  • Ri is Ci-C 6 alkyl, C 2 -C 6 alkyl ether, phenylC 0 -C 2 alkyl or (4- to 7-membered heterocycle)C 0 -C 2 alkyl, each of which is substituted with from 0 to 4 substituents independently chosen from oxo, halogen, hydroxy, and Ci-C 4 alkyl; and R 2 is hydrogen or Ci-C 4 alkyl.
  • variable "Ar,” in Formulas A, I, II and VII (and the various sub formulas thereof) is generally a 5- or 6-membered heteroaryl that contains at least one nitrogen atom, and is optionally substituted with one or more substituents independently chosen from R 3 , as described above.
  • Ar is a 6-membered heteroaryl, this group is
  • R 3 represents from O to 2 substituents independently chosen from halogen, Ci-C 4 alkyl, Ci-C 4 hydroxyalkyl, Ci-C 4 alkoxy, C 2 -C 4 alkyl ether, and mono- or di-(Ci-C 4 alkyl)amino.
  • One class of such compounds satisfies the Formula:
  • each R 9 is independently hydrogen or Ci-C 4 alkyl, or both Rg are taken together to form a spiro C 3 -C 8 cycloalkyl.
  • two substituents represented by R 3 are taken together to form an optionally-substituted 5- to 7-membered fused carbocyclic or heterocyclic ring.
  • ⁇ W ) ⁇ - ⁇ ⁇ represents a 5- or 6-membered carbocycle or a 5- or 6-membered heterocycle, each of which is substituted with from O to 4 substituents independently chosen from halogen, hydroxy, amino, Ci-C 6 alkyl, (C 3 -C 8 carbocycle)C 0 -C 4 alkyl, Ci-C 6 haloalkyl, Ci-C 6 alkoxy, Ci-C 6 alkyl ether, mono- or di-(Ci-C 6 alkyl)aminoC 0 -C 2 alkyl, and (4- to 7-membered heterocycle)C 0 -C 4 alkyl.
  • Ax is a 5-membered heteroaryl
  • R 3 represents 0 substituents or one substituent chosen from Ci-C 4 alkyl ⁇ e.g., methyl, ethyl or propyl), Ci-C 4 hydroxyalkyl and C 2 -C 4 alkyl ether. Certain such groups
  • Ar moieties include, for example:
  • R 3a is hydrogen, Ci-C 4 alkyl ⁇ e.g., methyl), Ci-C 4 hydroxyalkyl or C 2 -
  • R 4 moieties are indicated as "fused phenyl or 5- or 6-membered heterocyclic rings.” In such groups, the ring represented by R 4 is fused to the Z-containing ring, as in the following representative subformulas:
  • Yi, Y 2 , Y 3 and Y 4 are independently N, C, or CH and R 8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci- C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, (C 3 -C 8 cycloalkyl)C 0 -C 4 alkyl, Ci-C 6 alkoxy, and mono- or di-(Ci-C 6 alkyl)aminoC 0 -C 4 alkyl.
  • Y] is N and Y 2 , Y 3 and Y 4 are each CH; in other embodiments, Y 4 is N and Yi, Y 2 and Y 3 are each CH.
  • R 4 moieties are "a phenyl or 5- or 6-membered heteroaryl substituent.” In such groups, the ring represented by R 4 is pendant, as in the following representative subformulas:
  • Yi is N and Y 2 , Y 3 , Y 4 and Y 5 are each CH.
  • variable "n" in Formulas I, III, IV, VII, VIII and IX (as well as the subformulas thereof) is generally 0, 1 or 2. Within certain compounds, n is 0 (i.e., the ring
  • Formulas IHd-IIIh, IVd-IVh, VIIId-VIIIh and IXd-IXh illustrate certain amide- substituted aryl piperidines in which n is 0, 1 or 2, R 5a is hydrogen, Ci-C 6 alkyl, C 2 - C ⁇ alkenyl, C 2 -C 6 alkynyl or (C 3 -C 8 cycloalkyl)C 0 -C 4 alkyl and the remaining variables are as described above:
  • the group designated w is generally a heterocycle, as described above, that forms a spirocyclic system with the piperidine.
  • Representative such spirocyclic systems include, for example:
  • Yi is N and Y 2 , Y 3> Y 4 and Y 5 (if present) are each CH; in other embodiments, Y 4 is N and Yi, Y 2 , Y 3 and Y 5 (if present) are each CH.
  • R 5 (where present), R 6 and R 8 (where present) each represent O substituents within certain embodiments of the above Formulas.
  • amide-substituted aryl piperidines provided herein include, but are not limited to, those specifically described in the Examples below. It will be apparent that the specific compounds recited herein are representative only, and are not intended to limit the scope of the present invention. Further, as noted above, all compounds of the present invention may be present as a free acid or base or as a pharmaceutically acceptable salt, hydrate or ester.
  • compounds provided herein may be evaluated for certain pharmacological properties including, but not limited to, oral bioavailability (preferred compounds are orally bioavailable to an extent allowing for therapeutically effective doses of less than 140 mg/kg, preferably less than 50 mg/kg, more preferably less than 30 mg/kg, even more preferably less than 10 mg/kg, still more preferably less than 1 mg/kg and most preferably less than 0.1 mg/kg), toxicity (a preferred compound is nontoxic when a therapeutically effective amount is administered to a subject), side effects (a preferred compound produces side effects comparable to placebo when a therapeutically effective amount of the compound is administered to a subject), serum protein binding and in vitro and in vivo half-life (a preferred compound exhibits an in vivo half-life allowing for Q.I.D.
  • T. LD. dosing preferably T. LD. dosing, more preferably B. LD. dosing, and most preferably once- a-day dosing).
  • differential penetration of the blood brain barrier may be desirable. Routine assays that are well known in the art may be used to assess these properties, and identify superior compounds for a particular use. For example, assays used to predict bioavailability include transport across human intestinal cell monolayers, including Caco-2 cell monolayers. Penetration of the blood brain barrier of a compound in humans may be predicted from the brain levels of the compound in laboratory animals given the compound (e.g., intravenously). Serum protein binding may be predicted from albumin binding assays. Compound half-life is inversely proportional to the frequency of dosage of a compound. In vitro half- lives of compounds may be predicted from assays of microsomal half-life as described herein.
  • nontoxic compounds are nontoxic.
  • the term "nontoxic” as used herein shall be understood in a relative sense and is intended to refer to any substance that has been approved by the United States Food and Drug Administration (“FDA”) for administration to mammals (preferably humans) or, in keeping with established criteria, is susceptible to approval by the FDA for administration to mammals (preferably humans).
  • FDA United States Food and Drug Administration
  • a highly preferred nontoxic compound generally satisfies one or more of the following criteria: (1) does not substantially inhibit cellular ATP production; (2) does not significantly prolong heart QT intervals; (3) does not cause substantial liver enlargement, or (4) does not cause substantial release of liver enzymes.
  • a compound that does not substantially inhibit cellular ATP production is a compound that satisfies the criteria set forth in Example 6, herein.
  • cells treated as described in Example 6 with a 100 ⁇ M concentration of such a compound exhibit ATP levels that are at least 50% of the ATP levels detected in untreated cells.
  • such cells exhibit ATP levels that are at least 80% of the ATP levels detected in untreated cells.
  • a compound that does not significantly prolong heart QT intervals is a compound that does not result in a statistically significant prolongation of heart QT intervals (as determined by electrocardiography) in guinea pigs, minipigs or dogs upon administration of a dose that yields a serum concentration equal to the EC 50 or IC 50 for the compound.
  • a dose of 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 40 or 50 mg/kg administered parenterally or orally does not result in a statistically significant prolongation of heart QT intervals.
  • statically significant is meant results varying from control at the p ⁇ 0.1 level or more preferably at the p ⁇ 0.05 level of significance as measured using a standard parametric assay of statistical significance such as a student's T test.
  • a compound does not cause substantial liver enlargement if daily treatment of laboratory rodents (e.g., mice or rats) for 5-10 days with a dose that yields a serum concentration equal to the EC 50 or IC 50 for the compound results in an increase in liver to body weight ratio that is no more than 100% over matched controls. In more highly preferred embodiments, such doses do not cause liver enlargement of more than 75% or 50% over matched controls. If non-rodent mammals (e.g., dogs) are used, such doses should not result in an increase of liver to body weight ratio of more than 50%, preferably not more than 25%, and more preferably not more than 10% over matched untreated controls. Preferred doses within such assays include 0.01, 0.05. 0.1, 0.5, 1, 5, 10, 40 or 50 mg/kg administered parenterally or orally.
  • a compound does not promote substantial release of liver enzymes if administration of twice the minimum dose that yields a serum concentration equal to the EC 50 or IC 50 for the compound does not elevate serum levels of ALT, LDH or AST in laboratory rodents by more than 100% over matched mock-treated controls, hi more highly preferred embodiments, such doses do not elevate such serum levels by more than 75% or 50% over matched controls.
  • a compound does not promote substantial release of liver enzymes if, in an in vitro hepatocyte assay, concentrations (in culture media or other such solutions that are contacted and incubated with hepatocytes in vitro) that are equal to the EC 50 or IC 50 for the compound do not cause detectable release of any of such liver enzymes into culture medium above baseline levels seen in media from matched mock-treated control cells, hi more highly preferred embodiments, there is no detectable release of any of such liver enzymes into culture medium above baseline levels when such compound concentrations are five-fold, and preferably ten-fold the EC 50 or IC 50 for the compound.
  • certain preferred compounds do not inhibit or induce microsomal cytochrome P450 enzyme activities, such as CYP 1A2 activity, CYP2A6 activity, CYP2C9 activity, CYP2C19 activity, CYP2D6 activity, CYP2E1 activity or CYP3 A4 activity at a concentration equal to the EC 5O or IC 50 for the compound.
  • Certain preferred compounds are not clastogenic (e.g., as determined using a mouse erythrocyte precursor cell micronucleus assay, an Ames micronucleus assay, a spiral micronucleus assay or the like) at a concentration equal the EC 50 or IC 50 for the compound.
  • certain preferred compounds do not induce sister chromatid exchange (e.g., in Chinese hamster ovary cells) at such concentrations.
  • compounds provided herein may be isotopically-labeled or radiolabeled.
  • such compounds may have one or more atoms replaced by an atom of the same element having an atomic mass or mass number different from the atomic mass or mass number usually found in nature.
  • isotopes examples include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 15 N, 18 0, 17 0, 31 P, 32 P, 35 S, 18 F and 36 Cl.
  • substitution with heavy isotopes such as deuterium (i.e., 2 H) can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances.
  • Suitable protecting groups and methodology for protection and deprotection such as those described in Protecting Groups in Organic Synthesis by T. Greene, are well known. Compounds and intermediates requiring protection/deprotection will be readily apparent.
  • a compound provided herein may contain one or more asymmetric carbon atoms, so that the compound can exist in different stereoisomeric forms.
  • Such forms can be, for example, racemates or optically active forms.
  • All stereoisomers are encompassed by the present invention. Nonetheless, it may be desirable to obtain single enantiomers (i.e., optically active forms).
  • Standard methods for preparing single enantiomers include asymmetric synthesis and resolution of racemates. Resolution of racemates can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography using, for example a chiral HPLC column.
  • Compounds may be radiolabeled by carrying out their synthesis using precursors comprising at least one atom that is a radioisotope.
  • Each radioisotope is preferably carbon (e.g., 14 C), hydrogen (e.g., 3 H), sulfur (e.g., 35 S) or iodine (e.g., 125 I).
  • Tritium labeled compounds may also be prepared catalytically via platinum-catalyzed exchange in tritiated acetic acid, acid-catalyzed exchange in tritiated trifluoroacetic acid, or heterogeneous- catalyzed exchange with tritium gas using the compound as substrate.
  • certain precursors may be subjected to tritium-halogen exchange with tritium gas, tritium gas reduction of unsaturated bonds, or reduction using sodium borotritide, as appropriate.
  • Preparation of radiolabeled compounds may be conveniently performed by a radioisotope supplier specializing in custom synthesis of radiolabeled probe compounds.
  • compositions comprising one or more amide-substituted aryl piperidines provided herein, together with at least one physiologically acceptable carrier or excipient.
  • Pharmaceutical compositions may comprise, for example, one or more of water, buffers (e.g., sodium bicarbonate, neutral buffered saline or phosphate buffered saline), ethanol, mineral oil, vegetable oil, dimethylsulfoxide, carbohydrates (e.g., glucose, mannose, sucrose, starch, mannitol or dextrans), proteins, adjuvants, polypeptides or amino acids such as glycine, antioxidants, chelating agents such as EDTA or glutathione and/or preservatives.
  • other active ingredients may (but need not) be included in the pharmaceutical compositions provided herein.
  • compositions may be formulated for any appropriate manner of administration, including, for example, topical, oral (including, but not limited to, sublingual), nasal, rectal or parenteral administration.
  • parenteral as used herein includes subcutaneous, intradermal, intravascular (e.g., intravenous), intramuscular, spinal, intracranial, intrathecal and intraperitoneal injection, as well as any similar injection or infusion technique.
  • compositions suitable for oral use are preferred.
  • Such compositions include, for example, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs.
  • compositions of the present invention may be formulated as a lyophilizate.
  • Compositions intended for oral use may further comprise one or more components such as sweetening agents, flavoring agents, coloring agents and/or preserving agents in order to provide appealing and palatable preparations.
  • Tablets contain the active ingredient in admixture with physiologically acceptable excipients that are suitable for the manufacture of tablets.
  • excipients include, for example, inert diluents (e.g., calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate), granulating and disintegrating agents (e.g., corn starch or alginic acid), binding agents (e.g., starch, gelatin or acacia) and lubricating agents (e.g., magnesium stearate, stearic acid or talc).
  • inert diluents e.g., calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate
  • granulating and disintegrating agents e.g., corn starch or
  • the tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period.
  • a time delay material such as glyceryl monosterate or glyceryl distearate may be employed.
  • Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent (e.g., calcium carbonate, calcium phosphate or kaolin), or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin or olive oil).
  • an inert solid diluent e.g., calcium carbonate, calcium phosphate or kaolin
  • an oil medium e.g., peanut oil, liquid paraffin or olive oil.
  • Such excipients include suspending agents (e.g., sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia); and dispersing or wetting agents (e.g., naturally-occurring phosphatides such as lecithin, condensation products of an alkylene oxide with fatty acids such as polyoxyethylene stearate, condensation products of ethylene oxide with long chain aliphatic alcohols such as heptadecaethyleneoxycetanol, condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides such as polyethylene sorbitan monooleate).
  • suspending agents e.g., sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate,
  • Aqueous suspensions may also comprise one or more preservatives, such as ethyl or n- propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.
  • Oily suspensions may be formulated by suspending the active ingredient(s) in a vegetable oil (e.g., arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil such as liquid paraffin.
  • the oily suspensions may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol.
  • Sweetening agents such as those set forth above, and/or flavoring agents may be added to provide palatable oral preparations.
  • Such suspensions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
  • Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives.
  • a dispersing or wetting agent e.g., glycerol, glycerol, glycerol, glycerol, glycerol, glycerol, glycerin, glycerin, glycerin, glycerin, glycerin, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, glycerol, glycerol, glycerol, glycerol, glycerol, glycerol, glycerol, glycerol, glycerol
  • compositions may also be formulated as oil-in-water emulsions.
  • the oily phase may be a vegetable oil (e.g., olive oil or arachis oil), a mineral oil (e.g., liquid paraffin) or a mixture thereof.
  • Suitable emulsifying agents include naturally- occurring gums (e.g., gum acacia or gum tragacanth), naturally-occurring phosphatides (e.g., soy bean lecithin, and esters or partial esters derived from fatty acids and hexitol), anhydrides (e.g., sorbitan monoleate) and condensation products of partial esters derived from fatty acids and hexitol with ethylene oxide (e.g., polyoxyethylene sorbitan monoleate).
  • An emulsion may also comprise one or more sweetening and/or flavoring agents.
  • Syrups and elixirs may be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also comprise one or more demulcents, preservatives, flavoring agents and/or coloring agents.
  • Formulations for topical administration typically comprise a topical vehicle combined with active agent(s), with or without additional optional components. Suitable topical vehicles and additional components are well known in the art, and it will be apparent that the choice of a vehicle will depend on the particular physical form and mode of delivery.
  • Topical vehicles include water; organic solvents such as alcohols (e.g., ethanol or isopropyl alcohol) or glycerin; glycols (e.g., butylene, isoprene or propylene glycol); aliphatic alcohols (e.g., lanolin); mixtures of water and organic solvents and mixtures of organic solvents such as alcohol and glycerin; lipid-based materials such as fatty acids, acyl glycerols (including oils, such as mineral oil, and fats of natural or synthetic origin), phosphoglycerides, sphingolipids and waxes; protein-based materials such as collagen and gelatin; silicone-based materials (both non- volatile and volatile); and hydrocarbon-based materials such as microsponges and polymer matrices.
  • organic solvents such as alcohols (e.g., ethanol or isopropyl alcohol) or glycerin
  • glycols e.g., butylene, isoprene or prop
  • a composition may further include one or more components adapted to improve the stability or effectiveness of the applied formulation, such as stabilizing agents, suspending agents, emulsifying agents, viscosity adjusters, gelling agents, preservatives, antioxidants, skin penetration enhancers, moisturizers and sustained release materials.
  • stabilizing agents such as hydroxymethylcellulose or gelatin- microcapsules, liposomes, albumin microspheres, microemulsions, nanoparticles or nanocapsules.
  • a topical formulation may be prepared in a variety of physical forms including, for example, solids, pastes, creams, foams, lotions, gels, powders, aqueous liquids and emulsions.
  • Typical modes of delivery for topical compositions include application using the fingers; application using a physical applicator such as a cloth, tissue, swab, stick or brush; spraying (including mist, aerosol or foam spraying); dropper application; sprinkling; soaking; and rinsing. Controlled release vehicles can also be used.
  • a pharmaceutical composition may be prepared as a sterile injectible aqueous or oleaginous suspension.
  • the compound(s) provided herein, depending on the vehicle and concentration used, can either be suspended or dissolved in the vehicle.
  • Such a composition may be formulated according to the known art using suitable dispersing, wetting and/or suspending agents such as those mentioned above.
  • suitable dispersing, wetting and/or suspending agents such as those mentioned above.
  • the acceptable vehicles and solvents that may be employed are water, 1,3-butanediol, Ringer's solution and isotonic sodium chloride solution, hi addition, sterile, fixed oils may be employed as a solvent or suspending medium.
  • any bland fixed oil may be employed, including synthetic mono- or diglycerides.
  • fatty acids such as oleic acid find use in the preparation of injectible compositions, and adjuvants such as local anesthetics, preservatives and/or buffering agents can be dissolved in the vehicle.
  • compositions may also be formulated as suppositories (e.g., for rectal administration).
  • Such compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug.
  • Suitable excipients include, for example, cocoa butter and polyethylene glycols.
  • Pharmaceutical compositions may be formulated for release at a pre-determined rate. Instantaneous release may be achieved, for example, via sublingual administration (i.e., administration by mouth in such a way that the active ingredient(s) are rapidly absorbed via the blood vessels under the tongue rather than via the digestive tract).
  • Controlled release formulations may be administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at a target site.
  • a controlled release formulation comprises a matrix and/or coating that delays disintegration and absorption in the gastrointestinal tract (or implantation site) and thereby provides a delayed action or a sustained action over a longer period.
  • One type of controlled-release formulation is a sustained-release formulation, in which at least one active ingredient is continuously released over a period of time at a constant rate.
  • the therapeutic agent is released at such a rate that blood (e.g., plasma) concentrations are maintained within the therapeutic range, but below toxic levels, over a period of time that is at least 4 hours, preferably at least 8 hours, and more preferably at least 12 hours.
  • blood e.g., plasma
  • Such formulations may generally be prepared using well known technology and administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at the desired target site.
  • Carriers for use within such formulations are biocompatible, and may also be biodegradable; preferably the formulation provides a relatively constant level of modulator release. The amount of modulator contained within a sustained release formulation depends upon, for example, the site of implantation, the rate and expected duration of release and the nature of the condition to be treated or prevented.
  • Controlled release may be achieved by combining the active ingredient(s) with a matrix material that itself alters release rate and/or through the use of a controlled-release coating.
  • the release rate can be varied using methods well known in the art, including (a) varying the thickness or composition of coating, (b) altering the amount or manner of addition of plasticizer in a coating, (c) including additional ingredients, such as release- modifying agents, (d) altering the composition, particle size or particle shape of the matrix, and (e) providing one or more passageways through the coating.
  • the amount of modulator contained within a sustained release formulation depends upon, for example, the method of administration (e.g., the site of implantation), the rate and expected duration of release and the nature of the condition to be treated or prevented.
  • the matrix material which itself may or may not serve a controlled-release function, is generally any material that supports the active ingredient(s).
  • a time delay material such as glyceryl monosterate or glyceryl distearate may be employed.
  • Active ingredient(s) may be combined with matrix material prior to formation of the dosage form (e.g., a tablet).
  • active ingredient(s) may be coated on the surface of a particle, granule, sphere, microsphere, bead or pellet that comprises the matrix material. Such coating may be achieved by conventional means, such as by dissolving the active ingredient(s) in water or other suitable solvent and spraying.
  • additional ingredients are added prior to coating (e.g., to assist binding of the active ingredient(s) to the matrix material or to color the solution).
  • the matrix may then be coated with a barrier agent prior to application of controlled-release coating. Multiple coated matrix units may, if desired, be encapsulated to generate the final dosage form.
  • a controlled release is achieved through the use of a controlled release coating (i.e., a coating that permits release of active ingredient(s) at a controlled rate in aqueous medium).
  • the controlled release coating should be a strong, continuous film that is smooth, capable of supporting pigments and other additives, nontoxic, inert and tack-free.
  • Coatings that regulate release of the modulator include pH- independent coatings, pH-dependent coatings (which may be used to release modulator in the stomach) and enteric coatings (which allow the formulation to pass intact through the stomach and into the small intestine, where the coating dissolves and the contents are absorbed by the body).
  • pH dependent coatings include, for example, shellac, cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropylmethylcellulose phthalate, methacrylic acid ester copolymers and zein.
  • the coating is a hydrophobic material, preferably used in an amount effective to slow the hydration of the gelling agent following administration.
  • Suitable hydrophobic materials include alkyl celluloses (e.g., ethylcellulose or carboxymethylcellulose), cellulose ethers, cellulose esters, acrylic polymers (e.g., poly(acrylic acid), poly(methacrylic acid), acrylic acid and methacrylic acid copolymers, methyl methacrylate copolymers, ethoxy ethyl methacrylates, cyanoethyl methacrylate, methacrylic acid alkamide copolymer, poly(methyl methacrylate), polyacrylamide, ammonio methacrylate copolymers, aminoalkyl methacrylate copolymer, poly(methacrylic acid anhydride) and glycidyl methacrylate copolymers) and mixtures of the foregoing.
  • Representative aqueous dispersions of ethylcellulose include, for
  • AQUACOAT® FMC Corp., Philadelphia, PA
  • SURELEASE® Colorcon, Inc., West Point, PA
  • Representative acrylic polymers include, for example, the various EUDRAGIT® (Rohm America, Piscataway, NJ) polymers, which may be used singly or in combination depending on the desired release profile, according to the manufacturer's instructions.
  • Suitable plasticizers for alkyl celluloses include, for example, dibutyl sebacate, diethyl phthalate, triethyl citrate, tributyl citrate and triacetin.
  • Suitable plasticizers for acrylic polymers include, for example, citric acid esters such as triethyl citrate and tributyl citrate, dibutyl phthalate, polyethylene glycols, propylene glycol, diethyl phthalate, castor oil and triacetin.
  • Controlled-release coatings are generally applied using conventional techniques, such as by spraying in the form of an aqueous dispersion.
  • the coating may comprise pores or channels to facilitate release of active ingredient. Pores and channels may be generated by well known methods, including the addition of organic or inorganic material that is dissolved, extracted or leached from the coating in the environment of use.
  • pore-forming materials include hydrophilic polymers, such as hydroxyalkylcelluloses (e.g., hydroxypropylmethylcellulose), cellulose ethers, synthetic water-soluble polymers (e.g., polyvinylpyrrolidone, cross-linked polyvinylpyrrolidone and polyethylene oxide), water-soluble polydextrose, saccharides and polysaccharides and alkali metal salts.
  • a controlled release coating may include one or more orifices, which may be formed my methods such as those described in US Patent Nos. 3,845,770; 4,034,758; 4,077,407; 4,088,864; 4,783,337 and 5,071,607.
  • Controlled-release may also be achieved through the use of transdermal patches, using conventional technology (see, e.g., US Patent No. 4,668,232).
  • controlled release formulations may be found, for example, in US Patent Nos. 4,572,833; 4,587,117; 4,606,909; 4,610,870; 4,684,516; 4,777,049; 4,994,276; 4,996,058; 5,128,143; 5,202,128; 5,376,384; 5,384,133; 5,445,829; 5,510,119; 5,618,560; 5,643,604; 5,891,474; 5,958,456; 6,039,980; 6,143,353; 6,126,969; 6,156,342; 6,197,347; 6,387,394; 6,399,096; 6,437,000; 6,447,796; 6,475,493; 6,491,950; 6,524,615; 6,838,094; 6,905,709; 6,923,984; 6,923,988; and 6,911,217; each of which is hereby incorporated by reference for its teaching of the preparation of controlled release dosage forms
  • a compound provided herein may be conveniently added to food or drinking water (e.g., for administration to non-human animals including companion animals (such as dogs and cats) and livestock).
  • Animal feed and drinking water compositions may be formulated so that the animal takes in an appropriate quantity of the composition along with its diet. It may also be convenient to present the composition as a premix for addition to feed or drinking water.
  • Amide-substituted aryl piperidines provided herein are generally administered in a therapeutically effective amount.
  • Preferred systemic doses are no higher than 50 mg per kilogram of body weight per day (e.g., ranging from about 0.001 mg to about 50 mg per kilogram of body weight per day), with oral doses generally being about 5-20 fold higher than intravenous doses (e.g., ranging from 0.01 to 40 mg per kilogram of body weight per day).
  • the amount of active ingredient that may be combined with the carrier materials to produce a single dosage unit will vary depending, for example, upon the patient being treated and the particular mode of administration. Dosage units will generally contain from about 10 ⁇ g to about 500 mg of an active ingredient. Optimal dosages may be established using routine testing, and procedures that are well known in the art.
  • compositions provided herein may, but need not, further comprise one or more additional pharmaceutical agents, such as an anti-inflammatory agent, analgesic, anti-migraine agent, serotonin agonist, anti-hypertensive and/or anticonvulsant.
  • additional pharmaceutical agents such as an anti-inflammatory agent, analgesic, anti-migraine agent, serotonin agonist, anti-hypertensive and/or anticonvulsant.
  • Anti-inflammatory agents include, for example, non-steroidal anti-inflammatory drugs (NSAIDs), non-specific and cyclooxygenase-2 (COX-2) specific cyclooxgenase enzyme inhibitors, gold compounds, corticosteroids, methotrexate, leflunomide, cyclosporine A, EVI gold, minocycline, azathioprine, tumor necrosis factor (TNF) receptor antagonists, soluble TNF alpha receptor (etanercept), anti-TNF alpha antibodies (e.g., infliximab and adalimumab), anti-C5 antibodies, interleukin-1 (IL-I) receptor antagonists (e.g., anakinra or IL-I trap), IL-18 binding protein, CTLA4-Ig (e.g., abatacept), anti- human IL-6 receptor monoclonal antibody (e.g., tocilizumab), LFA-3-Ig fusion proteins (e.g., alef
  • NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen or naproxen sodium, diclofenac, combinations of diclofenac sodium and misoprostol, sulindac, oxaprozin, difiunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, sodium nabumetone, sulfasalazine, tolmetin sodium, and hydroxychloroquine.
  • COX cyclooxygenase
  • NSAIDs further include salicylates such as acetylsalicylic acid or aspirin, sodium salicylate, choline and magnesium salicylates, and salsalate, as well as corticosteroids such as cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, and prednisone.
  • salicylates such as acetylsalicylic acid or aspirin
  • sodium salicylate sodium salicylate
  • choline and magnesium salicylates sodium salicylate
  • salsalate sodium salicylate
  • corticosteroids such as cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, and prednisone.
  • analgesics for use in combination with CGRP receptor modulators are also anti-inflammatory agents, and are listed above.
  • Other such medications include narcotic agents which typically act at one or more opioid receptor subtypes (e.g., ⁇ , K and/or ⁇ ), preferably as agonists or partial agonists.
  • opioid receptor subtypes e.g., ⁇ , K and/or ⁇
  • Such agents include opiates, opiate derivatives and opioids, as well as pharmaceutically acceptable salts and hydrates thereof.
  • narcotic analgesics include, within preferred embodiments, alfentanil, alphaprodine, anileridine, bezitramide, buprenorphine, butorphanol, codeine, diacetyldihydromorphine, diacetylmorphine, dihydrocodeine, diphenoxylate, ethylmorphine, fentanyl, heroin, hydrocodone, hydromorphone, isomethadone, levomethorphan, levorphane, levorphanol, meperidine, metazocine, methadone, methorphan, metopon, morphine, nalbuphine, opium extracts, opium fluid extracts, powdered opium, granulated opium, raw opium, tincture of opium, oxycodone, oxymorphone, paregoric, pentazocine, pethidine, phenazocine, piminodine, propoxyphene, racemethorphan
  • narcotic analgesic agents include acetorphine, acetyldihydrocodeine, acetylmethadol, allylprodine, alphracetylmethadol, alphameprodine, alphamethadol, benzethidine, benzylmorphine, betacetylmethadol, betameprodine, betamethadol, betaprodine, clonitazene, codeine methylbromide, codeine-N-oxide, cyprenorphine, desomo ⁇ hine, dextromoramide, diampromide, diethylthiambutene, dihydromorphine, dimenoxadol, dimepheptanol, dimethylthiamubutene, dioxaphetyl butyrate, dipipanone, drotebanol, ethanol, ethylmethylthiambutene, etonitazene,
  • analgesic agents include, for example acetaminophen (paracetamol); aspirin and other NSAIDs described above; NR2B antagonists; bradykinin antagonists; anti-migraine agents; anticonvulsants such as oxcarbazepine and carbamazepine; antidepressants (such as TCAs, SSRIs, SNRIs, substance P antagonists, etc.); spinal blocks; pentazocine/naloxone; meperidine; levo ⁇ hanol; bupreno ⁇ hine; hydromo ⁇ hone; fentanyl; sufentanyl; oxycodone; oxycodone/acetaminophen, nalbuphine and oxymo ⁇ hone.
  • analgesic agents include CB2-receptor agonists, such as AM 1241, capsaicin receptor antagonists and compounds that bind to the ⁇ 2 ⁇ subunit of voltage-gated calcium channels, such as gabapentin and pregabalin.
  • anti-migraine agents for use in combination with a CGRP receptor modulator provided herein include caffeine; beta-adrenergic receptor antagonists such as timolol, propanolol, atenolol or nadolol; bradykinin- 1 receptor antagonists; adenosine Al receptor agonists; NMDA antagonists; NK-I receptor antagonists; opiate agonists; ergotamines; methysergide; calcium channel blockers such as flunarizine; selective serotonin uptake inhibitors such as fluoxetine, paroxetine, sertraline, duloxetine, escitalopram, or citalopram; 5-HTi agonists, such as sumatripan, naratriptan, zolmatriptan, eletriptan, almotriptan, frovatriptan, donitriptan and rizatriptan; inhibitors of nitric oxide synthesis; and cyclooxygenas
  • additional pharmaceutical agents include decongestants such as phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline, epinephrine, naphazoline, xylometazoline, propylhexedrine, or levo- desoxy-ephedrine; sedating or non-sedating antihistamines; and/or asthma treatments such as beta-adrenergic receptor agonists; leukotriene D 4 antagonists (e.g., montelukast or zafirlukast).
  • decongestants such as phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline, epinephrine, naphazoline, xylometazoline, propylhexedrine, or levo- desoxy-ephedrine
  • sedating or non-sedating antihistamines and/or asthma treatments such as beta
  • compositions may be packaged for treating conditions responsive to CGRP receptor modulation (e.g., treatment of pain, such as migraine or headache, or other disorder(s) recited herein).
  • Packaged pharmaceutical preparations generally comprise a container holding a therapeutically effective amount of a pharmaceutical composition as described above and instructions (e.g., labeling) indicating that the composition is to be used for treating a condition responsive to CGRP receptor modulation in a patient (e.g., pain, such as migraine or headache, or other disorder as indicated herein).
  • a packaged pharmaceutical preparation comprises one or more amide-substituted aryl piperidines provided herein and one or more additional agents in the same package, either in separate containers within the package or in the same container (i.e., as a mixture). Preferred mixtures are formulated for oral administration (e.g., as pills, capsules, tablets or the like).
  • the package comprises a label bearing indicia indicating that the components are to be taken together for the treatment of pain.
  • the present invention provides methods for treating a condition responsive to CGRP receptor modulation in a patient.
  • the patient may be afflicted with such a condition, or may be free of symptoms but considered at risk for developing such a condition.
  • a condition is "responsive to CGRP receptor modulation" if the condition or symptom(s) thereof are alleviated, attenuated, delayed or otherwise improved by modulation of CRP receptor activity.
  • such methods comprise administering to the patient a therapeutically effective amount of at least one amide- substituted aryl piperidine as provided herein.
  • Conditions responsive to CGRP receptor modulation include, for example pain (e.g., headache such as migraine), vasodilation (e.g., neurogenic vasodilation or in the treatment of shock, sepsis and hot flashes associated with menopause or hormone treatment), inflammation (e.g., neurogenic inflammation, inflammatory diseases of the joints such as arthritis, and airway inflammatory diseases such as asthma and chronic obstructive pulmonary disease), bronchial hyperreactivity, high blood pressure, hypotonia, tachycardia, diabetes (e.g., non-insulin-dependent diabetes mellitus), nausea, vomiting, diarrhea, loss of appetite, diabetic gastroparesis, polydipsis, thermal injury, cardiovascular diseases, morphine tolerance, skin diseases (e.g., dermatitis, psoriasis, thermal and radiation-induced skin damage such as sunburn), encephalitis, brain trauma, ischemia, epilepsy, neurodegenerative diseases, tinnitus, inflammatory bowel disease,
  • Headache that may be treated as described herein includes, for example, sinus, cluster (i.e., migranous neuralgia) and tension headaches, temporomandibular pain and maxillary sinus pain.
  • Migraine headache and other migraine symptoms may also be treated; for example, migraine headaches may be prevented by administration of a compound provided herein as soon as a pre-migrainous aura is experienced by the patient.
  • Other pain that may be treated using the CGRP receptor modulators provided herein may be chronic or acute and includes, but is not limited to, peripheral nerve- mediated pain (especially neuropathic pain, such as pain due to diabetes, postherpetic neuralgia, nerve injury, vulvodynia, root avulsions, painful traumatic neuropathy and painful polyneuropathy).
  • visceral pain e.g., pancreatitis, interstitial cystitis and renal colic
  • persistent hyperalgesia inflammatory pain, repetitive motion pain, carpel tunnel syndrome, perioperative pain, algesia, oral neuropathic pain, toothache (dental pain), denture pain, ocular pain, postherpetic neuralgia, diabetic neuropathy, chemotherapy- induced neuropathy, reflex sympathetic dystrophy, trigeminal neuralgia, bone and joint pain (e.g., pain associated with osteoarthritis), rheumatoid arthritis, myofascial pain (e.g., muscular injury and fibromyalgia), Guillain-Barre syndrome, meralgia paresthetica, burning-mouth syndrome and/or pain associated with nerve and root damage, including as pain associated with peripheral nerve disorders (e.g., nerve entrapment and brachial plexus avulsions, amputation,
  • peripheral nerve disorders e.g., nerve entrapment and
  • Additional neuropathic pain conditions include causalgia (reflex sympathetic dystrophy - RSD, secondary to injury of a peripheral nerve), neuritis (including, for example, sciatic neuritis, peripheral neuritis, polyneuritis, optic neuritis, postfebrile neuritis, migrating neuritis, segmental neuritis and Gombault's neuritis), neuronitis, neuralgias (e.g., those mentioned above, cervicobrachial neuralgia, cranial neuralgia, geniculate neuralgia, glossopharyngial neuralgia, migranous neuralgia, idiopathic neuralgia, intercostals neuralgia, mammary neuralgia, mandibular joint neuralgia, Morton's neuralgia, nasociliary neuralgia, occipital neuralgia, red neuralgia, Sluder's neuralgia, splenopalatine neuralg
  • Further pain conditions that may be treated include Charcot's pains, intestinal gas pains, ear pain, heart pain, muscle pain, eye pain, orofacial pain (e.g., odontalgia), abdominal pain, gynaecological pain (e.g., menstrual pain, dysmenorrhoea, pain associated with cystitis, labor pain, chronic pelvic pain, chronic prostitis and endometriosis), acute and chronic back pain (e.g., lower back pain), gout, scar pain, hemorrhoidal pain, dyspeptic pains, pain associated with angina, nerve root pain, "non-painful" neuropathies, complex regional pain syndrome, homotopic pain and heterotopic pain - including pain associated with carcinoma, often referred to as cancer pain (e.g., in patients with bone cancer), pain (and inflammation) associated with venom exposure (e.g., due to snake bite, spider bite, or insect sting) and trauma associated pain (e.g., post-surgical pain such as post
  • Additional pain conditions that may be treated as described herein include pain associated with respiratory disorders as described above, autoimmune diseases, immunodeficiency disorders, hot flashes, inflammatory bowel disease, gastroesophageal reflux disease (GERD), irritable bowel syndrome and/or inflammatory bowel disease.
  • amide-substituted aryl piperidines provided herein may be administered alone or in combination with one or more additional agents that are suitable for treating the disorder of interest.
  • the compound(s) and additional agent(s) may be present in the same pharmaceutical composition, or may be administered separately in either order.
  • Representative additional pharmaceutical agents for use in combination therapy include those indicated above administered by way of any means discussed above.
  • Treatment regimens may vary depending on the compound used and the particular condition to be treated. In general, a dosage regimen of 4 times daily or less is preferred, with 1 or 2 times daily particularly preferred. Dosage may be on an as-needed basis (e.g., the patient may self-administer upon the onset of symptoms). It will be understood, however, that the specific dose and treatment regimen for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex and diet of the patient, the time of administration, the route of administration, the rate of excretion, any drug combination and the severity of the particular disease undergoing therapy. Dosages are generally as described above; in general, the use of the minimum dose sufficient to provide effective therapy is preferred.
  • Patients may generally be monitored for therapeutic effectiveness using medical or veterinary criteria suitable for the condition being treated or prevented. Dosages and methods of administration of any additional agent(s) can be found, for example, in the manufacturer's instructions or in the Physician 's Desk Reference. In certain embodiments, combination administration results in a reduction of the dosage of the additional agent required to produce a therapeutic effect ⁇ i.e., a decrease in the minimum therapeutically effective amount). Thus, preferably, the dosage of additional agent in a combination or combination treatment method of the invention is less than the maximum dose advised by the manufacturer for administration of the agent without combination with a CGRP receptor modulator provided herein.
  • this dose is less than 3 Zt, even more preferably less than Vi, and highly preferably less than 1 A of the maximum dose, while most preferably the dose is less than 10% of the maximum dose advised by the manufacturer for administration of the agent(s) when administered without combination administration as described herein. It will be apparent that the dose of CGRP antagonist as provided herein needed to achieve the desired effect may similarly be reduced by the administration of the additional agent.
  • the present invention provides a variety of non- pharmaceutical in vitro and in vivo uses for the compounds provided herein.
  • such compounds may be labeled and used as probes for the detection and localization of CGRP receptor (in samples such as cell preparations or tissue sections, preparations or fractions thereof).
  • compounds provided herein that comprise a suitable reactive group may be used in photoaff ⁇ nity labeling studies of receptor binding sites, m addition, compounds provided herein may be used as positive controls in assays for receptor activity, as standards for determining the ability of a candidate agent to bind to CGRP receptor, or as radiotracers for positron emission tomography (PET) imaging or for single photon emission computerized tomography (SPECT). Such methods can be used to characterize CGRP receptors in living subjects.
  • a suitable reactive group such as an aryl carbonyl, nitro or azide group
  • a compound may be labeled using any of a variety of well known techniques (e.g., radiolabeled with a radionuclide such as tritium, as described herein), and incubated with a sample for a suitable incubation time (e.g., determined by first assaying a time course of binding). Following incubation, unbound compound is removed (e.g., by washing), and bound compound detected using any method suitable for the label employed (e.g., autoradiography or scintillation counting for radiolabeled compounds; spectroscopic methods may be used to detect luminescent groups and fluorescent groups).
  • a radionuclide such as tritium, as described herein
  • a matched sample containing labeled compound and a greater (e.g., 10-fold greater) amount of unlabeled compound may be processed in the same manner.
  • a greater amount of detectable label remaining in the test sample than in the control indicates the presence of CGRP receptor in the sample.
  • Detection assays, including receptor autoradiography (receptor mapping) of CGRP receptor in cultured cells or tissue samples may be performed as described by Kuhar in sections 8.1.1 to 8.1.9 of Current Protocols in Pharmacology (1998) John Wiley & Sons, New York. Also provided herein are methods for detecting CGRP receptor agonist or antagonist activity of a test compound.
  • Such methods generally employ cells that stably express CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2.
  • Suitable cells include HEK293 cells that are transfected with recombinant DNAs that encode CRLR, RAMP-I and the rat olfactory cyclic nucleotide gated channel subunits 1 and 2 (GenBank # X55519 and #U12623, respectively).
  • RAMP-I is human (e.g., GenBank #NM_005855) and the CRLR is human (e.g., GenBank #NM_005795).
  • the resulting cells can take up extracellular calcium via the rat olfactory cyclic nucleotide gated channel in response to increased intracellular cyclic AMP that results from CGRP receptor activation.
  • Such cells are then contacted with (a) a test compound; (b) CGRP receptor agonist
  • an indicator of intracellular calcium e.g., a membrane permeable calcium sensitivity dye such as Fluo-4 AM, which produces a fluorescent signal when bound to Ca ++ ).
  • an indicator of intracellular calcium e.g., a membrane permeable calcium sensitivity dye such as Fluo-4 AM, which produces a fluorescent signal when bound to Ca ++ .
  • a response indicative of intracellular calcium level in the contacted cells is measured. If the indicator produces a fluorescent response, typically the maximum response observed over a period of time (e.g., 4 minutes) is determined. This response is compared to a response detected in control cells that are treated identically to those described above, except the absence of test compound.
  • Compounds with antagonist activity reduce fluorescent signal detected in the presence of agonist and indicator.
  • Mass spectroscopy in the following Examples is Electrospray MS, obtained in positive ion mode using a Waters ZMD II Mass Spectrometer (Waters Corp.; Milford, MA), equipped with a Waters 600 pump (Waters Corp.), Waters 996 photodiode array detector (Waters Corp.), and a Gilson 215 autosampler (Gilson, Inc.; Middleton, WI). MassLynxTM version 4.0 software with OpenLynx processing is used for data collection and analysis (Waters Corp., Milford MA).
  • LCMS conditions are as follows:
  • This Example illustrates the preparation of representative intermediates useful in the synthesis of certain amide-substituted aryl piperidines.
  • 2-Chloro-l,3-dimethylimidazolinium chloride (272 mg, 1.61 mmol) is added portionwise to a stirred mixture of 2-chloro-4-methylthiazole-5-carboxylic acid (190 mg, 1.07 mmol), /ert-butyl 5-[l,l-dimethyl-2-(methylamino)ethyl]-lH-pyrazole-l-carboxylate (270 mg, 1.07 mmol), and TEA (0.45 ml, 3.21 mmol) in 10 ml OfCH 2 Cl 2 at 0 °C. After 10-15 minutes, water (10 ml) is added, and the layers are separated.
  • 2-Chloro-l,3-dimethylimidazolinium chloride (315 mg, 1.86 mmol) is added portionwise to a stirred mixture of 2-chloro-4-methylthiazole-5-carboxylic acid (220 mg, 1.24 mmol), 7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis- hydrochloride (278 mg, 1.24 mmol) and TEA (0.69 ml, 4.96 mmol) in 10 ml OfCH 2 Cl 2 at 0 °C. After 10-15 minutes, water (10 ml) is added, and the layers are separated.
  • 2-Chloro-l,3-dimethylimidazolinium chloride (2.5 g, 14.88 mmol) is added portionwise to a stirred mixture of 2-chloro-6-(methoxymethyl)-isonicotinic acid (2.0 g, 9.92 mmol), 7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis- hydrochloride (2.2 g, 9.92 mmol) and TEA (5.5 ml, 39.68 mmol) in 30 ml Of CH 2 Cl 2 at 0 °C. After 20 min, water (20 ml) is added, and the layers are separated.
  • 2-Chloro-l,3-dimethylimidazolinium chloride (323 mg, 1.91 mmol) is added portionwise to a stirred mixture of 2-chloroisonicotinic acid (200 mg, 1.27 mmol), 7,7- dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis-hydrochloride (285 mg, 1.27 mmol) and TEA (0.71 ml, 5.08 mmol) in 15 ml OfCH 2 Cl 2 at 0 °C. After 10 minutes, water (15 ml) is added, and the layers are separated. The aqueous layer is extracted once more with 15 ml Of CH 2 Cl 2 .
  • Potassium cyanate (8.7 g, 107 mmol) is added to a solution of tert-butyl 4-[(2- ethoxy-2-oxoethyl)amino]piperidine-l-carboxylate (30.5 g, 107 mmol) in water (600 ml).
  • the pH of the reaction mixture is adjusted to 4-5 with acetic acid, and the resulting reaction mixture is heated to 40 0 C for 16 h. After cooling to rt, the mixture is filtered and the solid dried in vacuo to give the title compound (with some inorganic salts) as a white solid which is used without further purification.
  • TFA 25 ml
  • tert-butyl 4-(2,4-dioxoimidazolidin-l- yl)piperidine-l-carboxylate 5 g
  • the excess TFA is removed in vacuo to give the title compound (with some inorganic salts) as a white solid which is used without further purification.
  • NaBH(OAc) 3 (25.28 g, 119.3 mmol) is added in portions and the mixture is stirred at rt for 1 hr before it is washed with 10 % Na 2 CO 3 solution (100 mL x 3) and brine (50 mL x 2). The organic phase is dried over Na 2 SO 4 . The solvent is removed in vacuo to afford the title compound as a red oil, which is used without further purification in subsequent steps.
  • tert-Butyl 4-[(2-nitrobenzyl)amino]piperidine-l-carboxylate (40 g) is dissolved in EtOH (200 mL) and then added 10 % Pd/C (3 g, 2.82 mmol). The mixture is hydrogenated at rt for 2 h. The mixture is passed through celite, the celite is washed with EtOH, and the combined solution is concentrated in vacuo to afford the title compound as a yellow oil, which is used without further purification in subsequent steps.
  • tert-Butyl (5Z)-5-[(dimethylamino)methylene]-3,3-dimethyl-4-oxopiperidine- 1 - carboxylate (6.4 g, 22.7 mmol) is dissolved in anhydrous MeOH (25 mL). Hydrazine (0.80 g, 24.93 mmol) is added and the resulting mixture is stirred at rt overnight. The solvent is removed in vacuo to afford the title compound as a viscous oil.
  • N-Boc-4-piperidone (43.8 g, 219.8 mmol) is added and the resulting mixture is stirred at rt for 1 h.
  • 2-Chloroethyl dimethylsulfonium iodide (50.0 g, 198.0 mmol) is added in portions and the resulting mixture is stirred at rt for 2 h.
  • t-BuOK (24.7 g, 219.8 mmol) in t-BuOH (350 mL) and the resulting mixture is allowed to stir at rt for 48 h.
  • reaction mixture is poured into H 2 O (2.5 L), and the resulting mixture is extracted with EtOAc (500 mL x 5) and dried over MgSO 4 .
  • EtOAc 500 mL x 5
  • the solvent is removed in vacuo, and silica gel column chromatography affords the title compound as a white solid.
  • tert-BvXy ⁇ 4-oxo-9-azadispiro[2.1.2.3]decane-9-carboxylate (1.0 g, 3.98 mmol) is dissolved in DCM (5 mL) and cooled to 0 °C. TFA (4.61 g, 40.4 mmol) is added and the resulting solution is stirred at 0 °C to rt for 2 h. Volatiles are removed in vacuo, and DCM (50 mL) is added. To this solution is added saturated NaHCO 3 aqueous solution (10 mL) and the mixture is stirred at 0 °C for 30 min. The organic phase is separated and the aqueous phase is extracted with DCM (30 mL x 4).
  • t-BuOK (24.7 g, 219.8 mmol) is dissolved in t-BuOH (350 mL) after heating at 60 0 C.
  • N-Boc-4-piperidone (43.8 g, 219.8 mmol) is added and the resulting mixture is stirred at rt for 1 h.
  • 2-Chloroethyldimethylsulfonium iodide (50.0 g, 198.0 mmol) is added in portions and the resulting mixture is stirred at rt for 2 h.
  • a solution of n-BuLi in hexane (1.6 M, 76.4 ml, 122 mmol) is added dropwise to a solution of diisopropylamine (17.2 ml, 122 mmol) in 250 ml of THF at -78 °C under N 2 .
  • a solution of 2-chloroquinoline (20 g, 122 mmol) in 50 ml of THF is added via a dropping funnel over 30 min.
  • a solution of 1- benzyl-4-piperidone (22.6 ml, 122 mmol) in 10 ml of THF is added to the brown reaction mixture, and stirring is continued for an additional 40 min.
  • EXAMPLE 2 SYNTHESIS OF REPRESENTATIVE AMIDE-SUBSTITUTED ARYL PIPERIDINES This Example illustrates the synthesis of certain representative amide-substituted aryl piperidines. All amide-substituted aryl piperidines in this Example are CGRP receptor antagonists, and all except Example 2E have an IC 50 (determined as described in Example 5) that is 2 micromolar or less (i.e., the concentration of such compounds that is required to provide a 50% decrease in the fluorescence response of cells exposed to one IC 50 of CGRP is 2 micromolar or less). A.
  • TFA (23.03 g, 0.202 mol) is added to 2-(3,5-dimethoxybenzyl)-l'- ⁇ 4-[(7,7- dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)carbonyl]pyridin-2- yl ⁇ spiro[isoindole-l,4'-piperidin]-3(2H)-one (160 mg, 0.264 mmol), and the resulting solution is refluxed overnight. The volatiles are removed in vacuo. DCM (10 mL) is added and to the resulting solution is added saturated NaHCO 3 solution (10 mL).
  • TFA (23.03 g, 0.202 mol) is added to 6'-(3,5-dimethoxybenzyl)-l- ⁇ 4-[(7,7- dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)carbonyl]pyridin-2- yl ⁇ spiro[piperidine-4,7'-pyi ⁇ olo[3,4-b]pyridin]-5'(6 ⁇ )-one (146 mg, 0.240 mmol), and the resulting solution is reflux ed overnight. The volatiles are removed in vacuo. DCM (100 mL) is added and to the resulting solution is added saturated NaHCO 3 solution (10 mL).
  • reaction mixture is diluted with H 2 O (10 mL) and the resulting mixture is extracted with DCM (15 mL * 3). The extracts are dried over MgSO 4 and solvent is removed in vacuo. Silica gel column chromatography affords the title compound as a yellow solid.
  • Example 5 This Example illustrates the preparation of CGRP receptor-transfected cells for use in CGRP receptor calcium mobilization assays (Example 5).
  • hRAMP-1 human receptor activity-modify protein 1
  • hCRLR human calcitonin receptor-like receptor
  • This plasmid is transfected into HEK293 cells that stably express recombinant rat olfactory cyclic nucleotide gated channel subunit 1 (GenBank # X55519) and rat olfactory cyclic nucleotide gated channel subunit 2 (GenBank #U12623), thus generating cells that take up extracellular calcium via the rat olfactory cyclic nucleotide gated channel in response to increased intracellular cyclic AMP.
  • a clonal cell line stably expressing human CGRP receptor is selected by neomycin resistance and by a calcium mobilization assay using calcium sensitive fluorescent dye as detailed below.
  • EXAMPLE 5 CALCIUM MOBILIZATION ASSAY This Example illustrates representative calcium mobilization assays for use in evaluating test compounds for agonist and antagonist activity at CGRP receptor.
  • Test compounds are tested in serial dilutions from 10 ⁇ M, 4 ⁇ M, 1 ⁇ M, or 0.1 ⁇ M stocks; for positive control cells, no test compound is added. The plates are incubated for an additional 1 hour at room temperature, and then transferred to a FLIPR384 instrument (Molecular Devices, Sunnyvale, CA) for calcium mobilization analysis.
  • FLIPR384 instrument Molecular Devices, Sunnyvale, CA
  • activating agonist 25 ⁇ L of KRH containing CGRP
  • the FLIPR into the plate
  • the final CGRP concentration is the EC 50 for the CGRP-mediated response in these cells (1.6 nM CGRP).
  • Pn negative control cells 25 ⁇ L of KRH without CGRP is added at this stage. Fluorescence is monitored for at least 4 minutes. The data is analyzed as follows. First, the average maximum relative fluorescent unit (RFU) response from the negative control wells (no CGRP) is subtracted from the maximum response detected for each of the other experimental wells. Second, average maximum RFU response is calculated for the agonist response to CGRP in the positive control wells (agonist wells). Then, percent inhibition for each compound tested is calculated using the equation:
  • KALEID AGRAPH software Synergy Software, Reading, PA
  • Antagonists of the CGRP receptor decrease the response induced by CGRP alone (in the absence of test compound) by at least about 20%, preferably by at least about 50%, and most preferably by at least 80%, at a concentration of 10 micromolar or less, preferably 4 micromolar or less, and more preferably 1 micromolar or less.
  • the IC 50 such antagonists is preferably below 1 micromolar, 100 nanomolar, 10 nanomolar or 1 nanomolar.
  • Similar assays are performed in the absence of added CGRP for the determination of agonist activity of the test compounds.
  • the ability of a test compound to act as an agonist of CGRP receptor is determined by measuring the fluorescence response elicited by the test compound as a function of compound concentration.
  • CGRP receptor antagonists that exhibit no detectable agonist activity elicit no detectable fluorescence at concentrations of 100 nanomolar, or 1, 4 or 10 micromolar.
  • Example 6 MDCK CYTOTOXICITY ASSAY This Example illustrates the evaluation of compound toxicity using a Madin Darby canine kidney (MDCK) cell cytotoxicity assay.
  • test compound 1 ⁇ L is added to each well of a clear bottom 96-well plate (PACKARD, Meriden, CT) to give final concentration of compound in the assay of 10 ⁇ M, 100 ⁇ M or 200 ⁇ M. Solvent without test compound is added to control wells.
  • MDCK cells ATCC no. CCL-34 (American Type Culture Collection, Manassas,
  • Confluent MDCK cells are trypsinized, harvested, and diluted to a concentration of 0.1 x 10 6 cells/mL with warm (37°C) medium (VITACELL Minimum Essential Medium Eagle, ATCC catalog # 30-2003). 100 ⁇ L of diluted cells is added to each well, except for five standard curve control wells that contain 100 ⁇ L of warm medium without cells. The plate is then incubated at 37 0 C under 95% O 2 , 5% CO 2 for 2 hours with constant shaking.
  • warm (37°C) medium VITACELL Minimum Essential Medium Eagle, ATCC catalog # 30-2003
  • mammalian cell lysis solution from the Packard ATP-LITE-M Luminescent ATP detection kit
  • 50 ⁇ L of mammalian cell lysis solution is added per well, the wells are covered with PACKARD TOPSEAL stickers, and plates are shaken at approximately 700 rpm on a suitable shaker for 2 min.
  • the ATP-LITE-M Luminescent ATP detection kit is generally used according to the manufacturer's instructions to measure ATP production in treated and untreated MDCK cells. PACKARD ATP LITE-M reagents are allowed to equilibrate to room temperature. Once equilibrated, the lyophilized substrate solution is reconstituted in 5.5 mL of substrate buffer solution (from kit). Lyophilized ATP standard solution is reconstituted in deionized water to give a 10 mM stock.
  • PACKARD substrate solution 50 ⁇ L is added to all wells, which are then covered, and the plates are shaken at approximately 700 rpm on a suitable shaker for 2 min.
  • a white PACKARD sticker is attached to the bottom of each plate and samples are dark adapted by wrapping plates in foil and placing in the dark for 10 min.
  • Luminescence is then measured at 22°C using a luminescence counter (e.g., PACKARD TOPCOUNT Microplate Scintillation and Luminescence Counter or TECAN SPECTRAFLUOR PLUS), and ATP levels calculated from the standard curve.
  • ATP levels in cells treated with test compound(s) are compared to the levels determined for untreated cells.
  • Cells treated with 10 ⁇ M of a preferred test compound exhibit ATP levels that are at least 80%, preferably at least 90%, of the untreated cells.
  • a 100 ⁇ M concentration of the test compound is used, cells treated with preferred test compounds exhibit ATP levels that are at least 50%, preferably at least 80%, of the ATP levels detected in untreated cells.

Abstract

Amide-substituted aryl piperidines derivatives of the following Formulas are provided:(Formulas), in which the variables are as described herein. Such compounds may be used to modulate calcitonin gene-related peptide (CGRP) receptor activity in vivo or in vitro, and are particularly useful in the treatment of conditions responsive to CGRP modulation in humans, domesticated companion animals and livestock animals, including headache, such as migraine. Pharmaceutical compositions and methods for using them to treat such disorders are provided, as are methods for using such compounds for receptor localization studies and various in vitro assays.

Description

AMIDE-SUBSTITUTED ARYL PIPERIDINES
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the benefit of U.S. Provisional Application 60/865,862, filed November 15, 2006, which provisional application is incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
This invention relates generally to amide-substituted aryl piperidines and related compounds, and to the use of such compounds to treat conditions responsive to calcitonin gene-related peptide (CGRP) receptor modulation. The invention further relates to the use of such compounds as reagents for the identification of other agents that bind to CGRP receptor, and as probes for the detection and localization of CGRP receptor.
BACKGROUND
Calcitonin gene-related peptide (CGRP) is a naturally occurring peptide that is widely distributed in both the peripheral and central nervous systems. This peptide is primarily localized in sensory afferent and central neurons, where it exerts a number of biological effects, including vasodilation. CGRP is released from trigeminal ganglia nerves after nerve activation and is a strong cerebral and dural vessel dilator. Accordingly, CGRP regulates blood flow to the brain and meninges (which are pain- sensitive), and is involved in the pathophysiology of conditions such as migraine and cluster headaches.
CGRP exerts its biological actions by binding to specific cell surface receptors (CGRP receptors), which are G-protein coupled and in turn activate intracellular adenylate cyclase. CGRP receptors generally have three discrete components: (1) a transmembrane calcitonin receptor-like receptor (CRLR or CLR); (2) a transmembrane receptor activity modifying protein type one (RAMPl) and (3) an intracellular receptor component protein (RCP), all of which are required in order to form a functional CGRP receptor complex. Evans et al. (1996) J Biol. Chem 40:31438-43. CGRP receptor antagonists inhibit the action of CGRP at such receptors, and are useful in the treatment of a variety of conditions associated with CGRP receptor modulation, including pain, headache and migraine.
There is thus a need in the art for CGRP antagonists. The present invention fulfills this need and provides further related advantages.
SUMMARY OF THE INVENTION
The present invention provides amide-substituted aryl piperidines that satisfy Formula A, Formula I or Formula II:
Figure imgf000003_0001
Formula A
Figure imgf000003_0002
Formula I Formula II
Within Formulas A, I and II,
^^= represents a single or double bond;
Figure imgf000003_0003
are each a 5- or 6-membered heterocycle that:
(a) comprises at least one ring nitrogen atom;
(b) is substituted with oxo;
(c) is optionally substituted with Ci-C4alkyl; and
(d) is:
(i) substituted with a phenyl or 5- or 6-membered heteroaryl substituent; or (ii) fused to a phenyl or 5- or 6-membered heterocyclic ring; each of which (i) and (ii) is optionally substituted and is preferably substituted with from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci- C6alkyl, C2-Cόalkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ci-C6alkoxy, and mono- or di-(C]-C6alkyl)aminoCo-C4alkyl; and wherein X is C, CH, or N;
Ar is a 5- or 6-membered heteroaryl, each of which is optionally substituted and each of which is preferably substituted with from 0 to 2 substituents independently chosen from R3; Z is C, CH, CH2, N or NH; n is 0, 1 or 2;
R] is C]-C8aminoalkyl, Ci-C8hydroxyalkyl, C2-C8alkyl ether, phenylC0-C6alkyl, or (4- to 12-membered heterocycle)C0-C6alkyl, each of which is optionally substituted and each of which is preferably substituted with from 0 to 6 substituents independently chosen from:
(i) halogen, hydroxy, cyano, amino, aminocarbonyl, aminosulfonyl and oxo; and (ii) C,-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, Ci-C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8carbocycle)Co-C4alkyl, mono- or di- (Ci- C6alkyl)aminoC0-C4alkyl, and (4- to 12-membered heterocycle)Co-C4alkyl, each of which is optionally substituted and each of which is preferably substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino and Ci-C4alkyl; and
R2 is hydrogen, C]-C6alkyl or Ci-C6haloalkyl; or Ri and R2 are taken together to form a 4- to 12-membered heterocycle that is optionally substituted and is preferably substituted with from 0 to 6 substituents independently chosen from:
(i) halogen, hydroxy, cyano, amino, aminocarbonyl, aminosulfonyl and oxo; and (ii) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, C]-C6haloalkyl, Ci-C6hydroxyalkyl, C2-C6alkyl ether, Ci-C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8cycloalkyl)C0-C4alkyl, mono- or di- (Ci-C6alkyl)aminoC0-C4alkyl, phenylC0-C4alkyl, (4- to 12-membered heterocycle)Co-C4alkyl, and spiro C3-C8cycloalkyl; each of which is optionally substituted and each of which is preferably substituted with from 0 to 6 substituents independently chosen from hydroxy, oxo, halogen, amino, aminocarbonyl, aminosulfonyl, Ci-C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-Qalkanoyl, Ci-C6alkylsulfonyl, d-C6alkylsulfinyl, Ci- C6alkoxycarbonyl, mono- or di-(Ci-C6alkyl)aminocarbonyl, and mono- or CU-(C1- C6alkyl)aminosulfonyl;
Each R3 is independently chosen from:
(i) halogen, hydroxy, cyano and amino; and (ii) Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-
C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, and (4- to 7-membered heterocycle)C0-C4alkyl; each of which is optionally substituted and each of which is preferably substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, Ci-C4alkyl, (C3-C8carbocycle)C0-C4alkyl and (4- to 10-membered heterocycle)Co-C4alkyl; or two substituents represented by R3 are taken together to form a fused 5- to 7- membered carbocycle or heterocycle that is optionally substituted and is preferably substituted with from 0 to 3 substituents independently chosen from halogen, hydroxy, amino, Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-Cόhaloalkyl, Ci-C6alkoxy, Ci- C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoCo-C2alkyl, and (4- to 7-membered heterocycle)C0-C4alkyl;
R4 represents:
(i) oxo;
(ii) a phenyl or 5- or 6-membered heteroaryl substituent; or (iii) a fused phenyl or 5- or 6-membered heterocyclic ring; each of which (ii) and (iii) is optionally substituted and is preferably substituted with from 0 to 4 substituents independently chosen from:
(a) halogen, hydroxy, cyano, amino and oxo; and
(b) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Q-Cehydroxyalkyl, Ci-C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8cycloalkyl)Co-C4alkyl, mono- or di- (C1-
C6alkyl)aminoC0-C4alkyl, and (4- to 10-membered heterocycle)C0-C4alkyl; each of which is optionally substituted and each of which is preferably substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino, aminocarbonyl, aminosulfonyl, Ci-C6alkyl, Ci-Cόalkoxy, Ci-C6alkylthio, Ci-C6alkanoyl, Ci- C6alkylsulfonyl, Ci-C6alkylsulfinyl, Ci-C6alkoxycarbonyl, mono- or CU-(C1- C6alkyl)aminocarbonyl, and mono- or di-(Ci-C6alkyl)aminosulfonyl;
R5 represents from 0 to 4 substituents independently chosen from Ci-C6alkyl, C2- C6alkenyl, C2-C6alkynyl and (C3-C8cycloalkyl)Co-C4alkyl; and R6 represents from O to 4 substituents independently chosen from halogen, hydroxy, amino, cyano, Ci-C6alkyl, Ci-C6haloalkyl, Ci-C6alkoxy and Ci-C6alkyl ether.
Within certain embodiments, the present invention provides amide-substituted aryl piperidines that satisfy Formula III or IV:
Figure imgf000006_0001
Formula III Formula IV
Within Formulas III and VI,
Figure imgf000006_0002
wherein X is N;
Z is C, CH, CH2, N or NH; ^^ represents a single or double bond; n is 0, 1 or 2;
A, B, D and E are each C, CH or N, such that exactly one or two of A, B, D and E are N;
T, U and V are independently C, CH, N, NH, S and O, such that at least one of T,
U and V is N; exactly one of T, U and V is CH; and
Figure imgf000006_0003
aromatic; and R3 represents from 0 to 3 substituents independently chosen from:
(i) halogen, hydroxy, cyano and amino; and
(ii) Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-C6haloalkyl, d-C6alkoxy, Ci- Qalkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, or (4- to 7-membered heterocycle)Co-C4alkyl; each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, Ci-C4alkyl, (C3-C8carbocycle)C0- C4alkyl and (4- to 10-membered heterocycle)C0-C4alkyl; and or two substituents represented by R3 are taken together to form a fused 5- to 7- membered carbocycle or heterocycle that is substituted with from 0 to 3 substituents independently chosen from halogen, hydroxy, amino, Ci-C6alkyl, (C3-C8carbocycle)C0- C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, C]-C6alkyl ether, mono- or di-(Ci- C6alkyl)aminoCo-C2alkyl, and (4- to 7-membered heterocycle)C0-C4alkyl; and the remaining variables are as described above.
Within certain embodiments, the present invention provides amide-substituted aryl piperidines that satisfy Formula V or VI:
Figure imgf000007_0001
Formula V Formula VI
Within Formulas V and VI,
A, B, D and E are each C, CH or N, such that exactly one or two of A, B, D and E are N; T, U and V are independently C, CH, N, NH, S and O, such that at least one of T,
U and V is N; exactly one of T, U and V is CH; and
Figure imgf000007_0002
R3 represents from 0 to 3 substituents independently chosen from:
(i) halogen, hydroxy, cyano and amino; and
(ii) Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, d-C6haloalkyl, Ci-C6alkoxy, Ci- Qalkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, or (4- to 7-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, Ci-C4alkyl, (C3-C8carbocycle)C0- C4alkyl and (4- to 10-membered heterocycle)C0-C4alkyl; and or two substituents represented by R3 are taken together to form a fused 5- to 7- membered carbocycle or heterocycle that is substituted with from 0 to 3 substituents independently chosen from halogen, hydroxy, amino, d-C6alkyl, (C3-C8carbocycle)Co- C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, C]-C6alkyl ether, mono- or di-(Ci- C6alkyl)aminoC0-C2alkyl, and (4- to 7-membered heterocycle)Co-C4alkyl; and the remaining variables are as described above. Also provided herein, within further aspects, are amide-substituted aryl piperidines that satisfy Formula VII: Formula VII
Figure imgf000008_0001
Within Formula VII:
Figure imgf000008_0002
wherein
X is C;
Z is C, CH, CH2, N or NH; ^^= represents a single or double bond; n is 0, 1 or 2; and the remaining variables are as described above.
Within certain embodiments, such amide-substituted aryl piperidines further satisfy Formula VIII or IX:
Figure imgf000008_0003
Formula VIII Formula IX Within Formulas VIII and IX:
Figure imgf000008_0004
wherein
X is C;
Z is C, CH, CH2, N or NH;
=^^ represents a single or double bond; n is 0, 1 or 2; and the remaining variables are as described above.
Within certain aspects, amide-substituted aryl piperidines provided herein are CGRP receptor modulators and exhibit an EC50 or IC50 of no greater than 10 micromolar, 5 micromolar, 2 micromolar, 1 micromolar, 500 nanomolar, 100 nanomolar, 50 nanomolar or 10 nanomolar in an assay for determination of CGRP receptor agonist or antagonist activity.
In preferred embodiments, CGRP receptor modulators provided herein are CGRP receptor antagonists; in certain embodiments, such antagonists exhibit no detectable CGRP receptor agonist activity above background at their EC50 for CGRP receptor.
Within certain aspects, amide-substituted aryl piperidines provided herein are labeled with a detectable marker (e.g., radiolabeled or fluorescein conjugated).
The present invention further provides pharmaceutical compositions comprising at least one amide-substituted aryl piperidine provided herein in combination with a physiologically acceptable carrier or excipient.
Methods are further provided for inhibiting agonist-induced CGRP receptor activity. Within certain embodiments, the inhibition takes place in vitro. Such methods comprise contacting a CGRP receptor with at least one CGRP receptor antagonist as described herein, under conditions and in an amount or concentration sufficient to detectably inhibit agonist-induced CGRP receptor activity. Within other such aspects, the CGRP receptor is in a patient. Such methods comprise contacting cells expressing a CGRP receptor in a patient with a CGRP receptor antagonist as described herein in an amount or concentration that would be sufficient to detectably inhibit agonist-induced CGRP receptor activity in cells expressing CGRP receptor in vitro. The present invention further provides methods for treating a condition responsive to CGRP receptor modulation in a patient, comprising administering to the patient a therapeutically effective amount of at least one amide-substituted aryl piperidine provided herein.
Within other aspects, methods are provided for treating migraine or other headache in a patient, comprising administering to a patient suffering from (or at risk for) headache a therapeutically effective amount of at least one amide-substituted aryl piperidine provided herein.
Within further aspects, the present invention provides methods for determining the presence or absence of CGRP receptor in a sample, comprising: (a) contacting a sample with an amide-substituted aryl piperidine provided herein under conditions that permit binding of the compound to CGRP receptor; and (b) detecting a signal indicative of a level of the compound bound to CGRP receptor.
In yet another aspect, the present invention provides methods of preparing the compounds disclosed herein, including the intermediates. Within other aspects, methods are provided for detecting CGRP receptor antagonist activity of a test compound, comprising: (i) contacting test cells that stably expresses CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2 with: (a) a test compound; (b) an indicator of intracellular or cytosolic calcium; and (c) CGRP receptor agonist; (ii) detecting a response indicative of intracellular or cytosolic calcium level in the contacted test cells; (iii) contacting matched control cells that stably expresses CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2 with: (a) an indicator of intracellular or cytosolic calcium; and (b) CGRP receptor agonist; (iv) detecting a response indicative of intracellular or cytosolic calcium level in the contacted control cells; and (iii) comparing the responses detected in (ii) and (iv).
These and other aspects of the present invention will become apparent upon reference to the following detailed description.
DETAILED DESCRIPTION
As noted above, the present invention provides amide-substituted aryl piperidines, which may be used in vitro or in vivo in a variety of contexts, as described herein. TERMINOLOGY
Compounds are generally described herein using standard nomenclature. For compounds having asymmetric centers, it should be understood that (unless otherwise specified) all of the optical isomers and mixtures thereof are encompassed. In addition, compounds with carbon-carbon double bonds may occur in Z- and E- forms, with all isomeric forms of the compounds being included in the present invention unless otherwise specified. If a compound exists in various tautomeric forms, a recited compound is not limited to any one specific tautomer, but rather is intended to encompass all tautomeric forms. Certain compounds are described herein using a general formula that includes variables {e.g., Z, Ar, Ri). Unless otherwise specified, each variable within such a formula is defined independently of any other variable, and any variable that occurs more than one time in a formula is defined independently at each occurrence.
The term "amide-substituted aryl piperidine" encompasses any compound that satisfies Formula A or Formula I or Formula II or Formula VII, and optionally further satisfies one or more additional formulas provided herein. This term further includes pharmaceutically acceptable salts, solvates {e.g., hydrates) and esters of such compounds.
A "pharmaceutically acceptable salt" of a compound recited herein is an acid or base salt that is suitable for use in contact with the tissues of human beings or animals without excessive toxicity or carcinogenicity, and preferably without irritation, allergic response, or other problem or complication. Specific pharmaceutically acceptable anions for use in salt formation include, but are not limited to, acetate, 2-acetoxybenzoate, ascorbate, benzoate, bicarbonate, bromide, calcium edetate, carbonate, chloride, citrate, dihydrochloride, diphosphate, ditartrate, edetate, estolate (ethylsuccinate), formate, fumarate, gluceptate, gluconate, glutamate, glycolate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroiodide, hydroxymaleate, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, pamoate, pantothenate, phenylacetate, phosphate, polygalacturonate, propionate, salicylate, stearate, subacetate, succinate, sulfamate, sulfanilate, sulfate, sulfonates including besylate (benzenesulfonate), camsylate (camphorsulfonate), edisylate (ethane- 1 ,2-disulfonate), esylate (ethanesulfonate) 2-hydroxyethylsulfonate, mesylate (methanesulfonate), triflate (trifluoromethanesulfonate) and tosylate (p-toluenesulfonate), tannate, tartrate, teoclate and triethiodide. Similarly, pharmaceutically acceptable cations for use in salt formation include, but are not limited to ammonium, benzathine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine, and metals such as aluminum, calcium, lithium, magnesium, potassium, sodium and zinc. Those of ordinary skill in the art will recognize further pharmaceutically acceptable salts for the compounds provided herein. In general, a pharmaceutically acceptable acid or base salt can be synthesized from a parent compound that contains a basic or acidic moiety by any conventional chemical method. Briefly, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, the use of nonaqueous media, such as ether, ethyl acetate, ethanol, methanol, isopropanol or acetonitrile, is preferred.
It will be apparent that each compound provided herein may, but need not, be formulated as a solvate (e.g., a hydrate) or non-covalent complex. In addition, the various crystal forms and polymorphs are within the scope of the present invention. Also provided herein are prodrugs of the compounds provided herein. A "prodrug" is a compound that may not fully satisfy the structural requirements of a formula provided herein, but is modified in vivo, following administration to a patient, to produce a compound within the scope of such formula. For example, a prodrug may be an acylated derivative of a compound as provided herein. Prodrugs include compounds wherein hydroxy, amine or sulfhydryl groups are bonded to any group that, when administered to a mammalian subject, cleaves to form a free hydroxy, amino or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to, acetate, formate, phosphate and benzoate derivatives of alcohol and amine functional groups within the compounds provided herein. Prodrugs of the compounds provided herein may be prepared by modifying functional groups present in the compounds in such a way that the modifications are cleaved in vivo to yield the parent compounds.
As used herein, the term "alkyl" refers to a straight or branched chain saturated aliphatic hydrocarbon. Alkyl groups include groups having from 1 to 8 carbon atoms (Cj- C8alkyl), from 1 to 6 carbon atoms (Ci-C6alkyl) and from 1 to 4 carbon atoms (Ci-
C4alkyl), such as methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, 2- pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl or 3-methylpentyl. In some instances, a substituent of an alkyl group is specifically indicated. For example, "Ci- C6aminoalkyl" refers to a Ci-C6alkyl that has at least one -NH2 substituent, and "Ci- Qhydroxyalkyl" refers to a Ci -Qalkyl substituted with at least one -OH. "C0-C4alkyl" refers to a single covalent bond (C0) or an alkylene group having 1, 2, 3 or 4 carbon atoms; "C0-C2alkyl" refers to a single covalent bond or a methylene or ethylene group.
"Alkylene" refers to a divalent alkyl group, as defined above. Ci-C4alkylene is an alkylene group having 1, 2, 3 or 4 carbon atoms. "Alkenyl" refers to straight or branched chain alkene groups, which comprise at least one unsaturated carbon-carbon double bond. Alkenyl groups include C2-C8alkenyl, C2-C6alkenyl and C2-C4alkenyl groups, which have from 2 to 8, 2 to 6 or 2 to 4 carbon atoms, respectively, such as ethenyl, allyl or isopropenyl. "Alkynyl" refers to straight or branched chain alkyne groups, which have one or more unsaturated carbon-carbon bonds, at least one of which is a triple bond. Alkynyl groups include C2-C8alkynyl, C2-C6alkynyl and C2-C4alkynyl groups, which have from 2 to 8, 2 to 6 or 2 to 4 carbon atoms, respectively.
A "cycloalkyl" is a saturated or partially saturated cyclic group in which all ring members are carbon, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and partially saturated variants thereof. Certain cycloalkyl groups are C3-C8cycloalkyl, in which the ring contains from 3 to 8 ring members, all of which are carbon. A "(C3- C8cycloalkyl)C0-C4alkyl" is a C3-C8cycloalkyl group linked via a single covalent bond or a Ci-C4alkylene group. A "spiro C3-C8cycloalkyl" is a C3-C8cycloalkyl substituent of a ring attached so as to form a spiro ring system. For example, if R7 in the group designated: is a spiro cyclopropyl ring, one resulting ring system is
Figure imgf000013_0001
By "alkoxy," as used herein, is meant an alkyl group attached via an oxygen bridge {i.e., -O-alkyl). Alkoxy groups include Ci-C6alkoxy and Ci-C4alkoxy groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively. Methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy, neopentoxy, hexoxy, 2-hexoxy, 3-hexoxy and 3-methylpentoxy are representative alkoxy groups.
Similarly, "alkylthio" refers to an alkyl group as attached via a sulfur bridge {i.e., - S-alkyl). Alkylthio groups include Ci-C6alkylthio and Ci-C4alkylthio groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively.
"Alkylsulfinyl" refers to groups of the formula -(SO)-alkyl, in which the sulfur atom is the point of attachment. Alkylsulfinyl groups include Ci-C6alkylsulfinyl and Ci- C4alkylsulfmyl groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively. "Alkylsulfonyl" refers to groups of the formula -(SO2)-alkyl, in which the sulfur atom is the point of attachment. Alkylsulfonyl groups include Ci-C6alkylsulfonyl and Ci- C4alkylsulfonyl groups, which have from 1 to 6 or from 1 to 4 carbon atoms, respectively. The term "(Ci-C6alkylsulfonyl)C0-C4alkyl refers to a Ci-C6alkylsulfonyl that is linked via a single covalent bond or a Ci-C4alkylene group {i.e., -(C0-C4alkyl)-(SO2)-(Ci-C6alkyl)). The term "alkoxycarbonyl" refers to an alkoxy group linked via a carbonyl {i.e., a group having the general structure -C^CO-O-alkyl). Alkoxycarbonyl groups include Ci- C8, Ci-C6 and Ci-C4alkoxycarbonyl groups, which have from 1 to 8, 6 or 4 carbon atoms, respectively, in the alkyl portion of the group. "Cialkoxycarbonyl" refers to -C(=O)-O- CH3. The term "alkanoyl" refers to an acyl group {e.g., -(C=O)-alkyl), in which carbon atoms are in a linear or branched alkyl arrangement and where attachment is through the carbon of the keto group. Alkanoyl groups have the indicated number of carbon atoms, with the carbon of the keto group being included in the numbered carbon atoms. For example a C2alkanoyl group is an acetyl group having the formula -(C=O)CH3. Alkanoyl groups include, for example, C]-C8alkanoyl, Ci-C6alkanoyl and Ci-C4alkanoyl groups, which have from 1 to 8, from 1 to 6 or from 1 to 4 carbon atoms, respectively. "Cialkanoyl" refers to -(C=O)H.
"Alkanoylamino," as used herein, refers to an alkanoyl group linked via a nitrogen bridge (i.e., a group having the general structure -N(R)-C(=O)-alkyl), in which R is hydrogen or Ci-C6alkyl. Alkanoylamino groups include, for example, (Ci- C6alkanoyl)amino groups, which have from 1 to 6 carbon atoms within the alkanoyl group.
"Alkylamino" refers to a secondary or tertiary amine that has the general structure -NH-alkyl or -N(alkyl)(alkyl), wherein each alkyl is selected independently from alkyl, cycloalkyl and (cycloalkyl)alkyl groups. Such groups include, for example, mono- and di- (Ci-C8alkyl)amino groups, in which each Ci-C8alkyl may be the same or different, as well as mono- and di-(Ci-C6alkyl)amino groups and mono- and di-(Ci-C4alkyl)amino groups. "Alkylaminoalkyl" refers to an alkylamino group linked via an alkylene moiety (i.e., a group having the general structure -alkylene-NH-alkyl or -alkylene-
N(alkyl)(alkyl)) in which each alkyl is selected independently from alkyl, cycloalkyl and (cycloalkyl)alkyl groups. Alkylaminoalkyl groups include, for example, mono- and di- (Ci-C8alkyl)aminoCi-C6alkyl, and mono- and di-(Ci-C6alkyl)aminoCi-C4alkyl. "Mono- or di-(C]-C6alkyl)aminoC0-C4alkyl" refers to a mono- or di-(Ci-Cόalkyl)amino group linked via a single covalent bond or a Ci-C4alkylene group. The following are representative alkylaminoalkyl groups:
Figure imgf000014_0001
It will be apparent that the definition of "alkyl" as used in the terms "alkylamino" and "alkylaminoalkyl" differs from the definition of "alkyl" used for all other alkyl- containing groups, in the inclusion of cycloalkyl and (cycloalkyl)alkyl groups (e.g., (C3- C7cycloalkyl)C0-C2alkyl).
The term "aminocarbonyl" refers to an amide group (i.e., -C(=O)NH2). The term "mono- or di-(Ci-C6alkyl)aminocarbonyl" refers to groups of the formula -(C=O)-N(R)2, in which the carbonyl is the point of attachment, one R is Ci-C6alkyl and the other R is hydrogen or an independently chosen C!-C6alkyl. Similarly, "aminosulfonyl" refers to a sulfonamide group (i.e., -SO2NH2). The term "mono- or di-(Ci-C6alkyl)aminosulfonyl" refers to groups of the formula -(SO2)- N(R)2, in which the sulfonyl is the point of attachment, one R is C]-C6alkyl and the other R is hydrogen or an independently chosen Ci-C6alkyl. The term "halogen" refers to fluorine, chlorine, bromine or iodine.
The term "oxo" is used herein to refer to an oxygen substituent of a carbon atom that results in the formation of a carbonyl group (C=O). An oxo group that is a substituent of a nonaromatic carbon atom results in a conversion Of-CH2- to -C(=0)-. An oxo group that is a substituent of an aromatic carbon atom results in a conversion of -CH- to - C(=0)- and may result in a loss of aromaticity.
A "haloalkyl" is an alkyl group that is substituted with 1 or more independently chosen halogens (e.g., "Ci-Cshaloalkyl" groups have from 1 to 8 carbon atoms; "C]- C6haloalkyl" groups have from 1 to 6 carbon atoms). Examples of haloalkyl groups include, but are not limited to, mono-, di- or tri-fluoromethyl; mono-, di- or tri- chloromethyl; mono-, di-, tri-, tetra- or penta-fluoroethyl; mono-, di-, tri-, tetra- or penta- chloroethyl; and 1,2,2,2-tetrafluoro-l-trifluoromethyl-ethyl. Typical haloalkyl groups are trifiuoromethyl and difluoromethyl.
A dash ("-") that is not between two letters or numbers is used to indicate a point of attachment for a substituent. For example, -C(=0)NH2 is attached through the carbon atom.
A "carbocycle" or "carbocyclic group" comprises at least one ring formed entirely by carbon-carbon bonds (referred to herein as a carbocyclic ring), and does not contain a heterocycle. Unless otherwise specified, each ring within a carbocycle may be independently saturated, partially saturated or aromatic, and is optionally substituted as indicated. A carbocycle generally has from 1 to 3 fused, pendant or spiro rings; carbocycles within certain embodiments have one ring or two fused rings. Typically, each ring contains from 3 to 8 ring members (i.e., C3-C8); rings having from 5 to 7 ring members are recited in certain embodiments. Carbocycles comprising fused, pendant or spiro rings typically contain from 9 to 14 ring members. Certain representative carbocycles are cycloalkyl as described above. Other carbocycles are aryl (i.e., contain at least one aromatic carbocyclic ring, with or without one or more additional aromatic and/or cycloalkyl rings). Such aryl carbocycles include, for example, phenyl, naphthyl (e.g., 1 -naphthyl and 2-naphthyl), fluorenyl, indanyl and 1,2,3,4-tetrahydronaphthyl. Certain 3- to 8-membered carbocycles are linked via a single covalent bond or Ci- C4alkylene; and are accordingly designated "(C3-C8carbocycle)C0-C4alkyl." Such groups include, for example, aryl and arylalkyl moieties (such as phenyl, indanyl, and groups in which either of the foregoing is linked via Ci-C4alkylene; for example, a phenylCo-C4alkyl refers to a phenyl group linked via a single covalent bond or Ci-C4alkylene) and (cycloalkyl)alkyl moieties as described above.
A "heterocycle" (also referred to herein as a "heterocyclic group") has from 1 to 3 fused, pendant or spiro rings, at least one of which is a heterocyclic ring (i.e., one or more ring atoms is a heteroatom independently chosen from oxygen, sulfur and nitrogen, with the remaining ring atoms being carbon). Typically, a heterocyclic ring comprises 1, 2, 3 or 4 heteroatoms; within certain embodiments each heterocyclic ring has 1 or 2 heteroatoms per ring. Each heterocyclic ring generally contains from 4 to 8 ring members (rings having from 4 or 5 to 7 ring members are recited in certain embodiments) and heterocycles comprising fused, pendant or spiro rings typically contain from 9 to 14 ring members. Certain heterocycles comprise a sulfur atom as a ring member; in certain embodiments, the sulfur atom is oxidized to SO or SO2. Heterocycles may be optionally substituted with a variety of substituents, as indicated. Certain heterocycles are 4- to 10- membered or 4- to 10-membered and comprise one or two rings; in certain embodiments, such heterocycles are monocyclic (e.g., 4- to 8-membered, 5- to 8-membered, 4- to 7- membered, or 5- or 6-membered).
Certain heterocycles are heteroaryl groups (i.e., at least one heterocyclic ring within the group is aromatic), such as a 5- or 6-membered heteroaryl (e.g., thienyl, imidazolyl, pyridyl or pyrimidyl). Other heterocycles are heterocycloalkyl groups (i.e., do not comprise an aromatic heterocyclic ring). If so designated, a heterocycle may be linked by a single covalent bond or an alkylene group; for example, "(4- to 12-membered heterocycle)Co-C4alkyl" refers to a 4- to 12-membered heterocycle having 1 or 2 rings, which is linked via a single covalent bond or via an alkylene group having from 1 to 6 carbons. "(4- to 7-membered heterocycle)Co-C4alkyl" refers to a 4- to 7-membered heterocyclic ring that is linked via a single covalent bond or an alkylene group having from 1 to 4 carbons, methylene or ethylene linker. When substituted, it will be apparent that substituent(s) may be attached on the ring and/or the alkylene linker.
Certain heteroaryls are indicated in various Formulas with the following notation:
Figure imgf000017_0001
in which the variables are as described herein, such that the ring retains
aromaticity. It will be apparent, for example, that -K BO Dt R3 an(j
Figure imgf000017_0002
interchangeable notations. A "substituent," as used herein, refers to a molecular moiety that is covalently bonded to an atom within a molecule of interest. For example, a "ring substituent" may be a moiety such as a halogen, alkyl group, haloalkyl group or other group discussed herein that is covalently bonded to an atom (such as a carbon or nitrogen atom) that is a ring member. The term "substitution" refers to replacing a hydrogen atom in a molecular structure with a substituent as described above, such that the valence on the designated atom is not exceeded, and such that a chemically stable compound (i.e., a compound that can be isolated, characterized, and tested for biological activity) results from the substitution.
Groups that are "optionally substituted" are unsubstituted or are substituted by other than hydrogen at one or more available positions, typically 1, 2, 3, 4 or 5 positions, by one or more suitable groups (which may be the same or different). Optional substitution is also indicated by the phrase "substituted with from 0 to X substituents," where X is the maximum number of possible substituents. Certain optionally substituted groups are substituted with from 0 to 2, 3 or 4 independently selected substituents (i.e., are unsubstituted or substituted with up to the recited maximum number of substituents).
"CGRP receptor," as used herein, refers to any functional CGRP receptor complex, comprising a ligand binding protein CRLR and a chaperone protein RAMPl . In general, such a receptor complex further comprises a coupling protein RCP. In certain embodiments, each complex component is human: human CRLR includes, for example, the polypeptide having the sequence provided at Genbank accession number NM_005795, and human RAMPl includes, for example, the polypeptide having the sequence provided at Genbank accession number NM 005855. One or more components of the receptor complex may be expressed endogenously; in certain embodiments it is convenient to use cells that express CRLR and RAMPl recombinantly, with RCP expressed endogenously. Alternatively, any of a variety of cells such as the human neuroblastoma cell line SK-N- MS (DSMZ, Braunschweig, Germany), which naturally express functional CGRP receptor, may be a convenient source for CGRP receptor and receptor-containing membrane preparations. The term "CGRP receptor agonist" refers to a compound that binds CGRP receptor and induces signal transduction mediated by CGRP receptor. CGRP receptor agonists include, for example, CGRP (e.g., human CGRP), as well as peptide portions or variants thereof that bind CGRP receptor and retain CGRP-receptor activating activity.
A "CGRP receptor antagonist" is a compound that detectably inhibits signal transduction mediated by CGRP receptor. Such inhibition may be determined using the representative calcium mobilization assay provided in Example 5. Preferred CGRP receptor antagonists have an IC50 of 10 μM or less in this assay, more preferably 5 μM or less, and still more preferably 1 μM or less, 500 nM or less, or 100 nM or less. Certain CGRP receptor antagonists exhibit minimal agonist activity (i.e., they induce an increase in the basal activity of CGRP receptor that is less than 5% of the increase that would be induced by one EC50 of the peptide agonist CGRP) and more preferably exhibit no detectable agonist activity within the assay described in Example 5. CGRP receptor antagonists for use as described herein are generally non-toxic. CGRP receptor antagonists include neutral antagonists and inverse agonists. A "neutral antagonist" of CGRP receptor is a compound that inhibits the activity of
CGRP receptor agonist at CGRP receptor, but when present at its IC50 (determined using the calcium mobilization assay as described in Example 5 does not significantly change the basal activity of the receptor. The effect on the basal activity may be determined using the calcium mobilization assay described in Example 5 performed in the absence of agonist. Preferably, within such an assay, CGRP receptor activity is reduced by no more than 10%, more preferably by no more than 5%, and even more preferably by no more than 2%; most preferably, there is no detectable reduction in activity. Neutral antagonists may, but need not, also inhibit the binding of agonist to CGRP receptor.
An "inverse agonist" of CGRP receptor is a compound that reduces the activity of CGRP receptor below its basal activity level in the absence of activating concentrations of agonist. Inverse agonists may also inhibit the activity of agonist at CGRP receptor, and/or may inhibit binding of CGRP receptor agonist to CGRP receptor. The reduction in basal activity of CGRP receptor produced by an inverse agonist may be determined from a calcium mobilization assay, such as the assay of Example 5. A "therapeutically effective amount" (or dose) is an amount that, upon administration to a patient, results in a discernible patient benefit (e.g., provides detectable relief from a condition being treated). Such relief may be detected using any appropriate criteria. A therapeutically effective amount or dose generally results in a concentration of compound(s) in a body fluid (such as blood, plasma, serum, CSF, synovial fluid, lymph, cellular interstitial fluid, tears or urine) that is sufficient to result in detectable alteration in CGRP receptor -mediated signal transduction (using an assay provided herein). The discernible patient benefit may be apparent after administration of a single dose, or may become apparent following repeated administration of the therapeutically effective dose according to a predetermined regimen, depending upon the indication for which the compound is administered.
A "patient" is any individual treated with an amide-substituted aryl piperidine as provided herein. Patients include humans, as well as other animals such as companion animals (e.g., dogs and cats) and livestock; in certain embodiments, human patients are preferred. Patients may be experiencing one or more symptoms of a condition responsive to CGRP receptor modulation or may be free of such symptom(s) (i.e., treatment may be prophylactic in a patient considered to be at risk for the development of such symptoms).
AMIDE-SUBSTITUTED ARYL PIPERIDINES As noted above, the present invention provides amide-substituted aryl piperidines that satisfy any one of Formulas A, I, II, or VII, above. Such amide-substituted aryl piperidines may be used in a variety of contexts, including in the treatment of conditions responsive to CGRP receptor modulation. Such compounds may also be used within in vitro assays (e.g., assays for CGRP receptor activity), as probes for the detection and localization of CGRP receptor and within assays to identify other CGRP receptor antagonists.
Certain amide-substituted aryl piperidines of Formula A or Formula I or Formula II further satisfy one of Formulas III- VI, as described above. Certain amide-substituted aryl piperidines of Formula VII further satisfy Formula VIII of Formula IX, as described above.
Within certain embodiments of Formulas A and I - IX, the group designated:
Figure imgf000020_0001
, wherein:
R7 represents from 0 to 4 substituents independently chosen from: (i) halogen, hydroxy, amino and oxo; and (ii) Ci-Cόalkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Ci- C6alkoxycarbonyl, C2-C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3- C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Cp C4alkyl) amino; and
Figure imgf000020_0002
are each a 5- or 6-membered heterocycle (e.g., a 5-membered heteroaryl) that is optionally substituted with Ci-C4alkyl.
Certain such compounds satisfy one or more of Formulas Ia - IXa:
Figure imgf000020_0003
Formula Ia Formula Ha
Figure imgf000020_0004
Formula Ilia Formula FVa
Figure imgf000020_0005
Formula Va Formula Via
Figure imgf000021_0002
Formula Vila
Figure imgf000021_0001
Figure imgf000021_0003
Formula IXa
Within further such embodiments of Formulas Ia - IXa, the group designated:
Figure imgf000021_0004
Certain such compounds satisfy ( one or more of Formulas Ib - IXb:
Figure imgf000021_0005
Formula lib
Figure imgf000021_0007
Formula IVb
Figure imgf000021_0006
Figure imgf000021_0008
Formula Vb Formula VIb
Figure imgf000022_0001
Formula VIIb Formula VIIIb
Figure imgf000022_0002
Formula IXb
Representative R7 moieties in the above formulas include methyl (e.g., in certain compounds, R7 represents two methyl substituents, such as gem-dimethyl) and spiro C3- C6cycloalkyl groups. Within certain such compounds, the group designated:
Figure imgf000022_0003
, wherein each R9 is independently hydrogen or Ci-
C4alkyl, or both R9 are taken together to form a spiro C3-C8cycloalkyl, and Ri0 is hydrogen
or Ci-C4alkyl. Certain such groups include:
Figure imgf000022_0004
an(j m which m is an integer ranging from 1 to 4.
Within other embodiments of Formulas A and I - IX, the group designated:
-
Figure imgf000022_0005
wherein R7 is as described above; o is 0, 1 or 2 and M is NH, O, S, SO, SO2 or CH2. Certain such compounds satisfy one or more of Formulas Ic IXc:
Figure imgf000022_0006
Formula Ic Formula Hc
Figure imgf000023_0001
Formula IHc Formula IVc
Figure imgf000023_0002
Formula Vc Formula VIc
Figure imgf000023_0003
Formula VIIc Formula VIIIc
Figure imgf000023_0004
Formula FVc
Representative R7 moieties within such compounds include, for example hydroxy, amino, oxo, Ci-Cόalkyl, Ci-C6hydroxyalkyl, C]-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, (Ci-C6alkanoyl)amino, Ci-C6alkoxycarbonyl, C2-C6alkyl ether, phenyl and 4- to 7- membered heterocycles. Within certain such compounds, o is 1 (i.e.,
Figure imgf000023_0005
and/or R7 represents two methyl substituents.
Within other embodiments of Formulas A and I - IX, Ri is Ci-C6alkyl, C2-C6alkyl ether, phenylC0-C2alkyl or (4- to 7-membered heterocycle)C0-C2alkyl, each of which is substituted with from 0 to 4 substituents independently chosen from oxo, halogen, hydroxy, and Ci-C4alkyl; and R2 is hydrogen or Ci-C4alkyl. The variable "Ar," in Formulas A, I, II and VII (and the various sub formulas thereof) is generally a 5- or 6-membered heteroaryl that contains at least one nitrogen atom, and is optionally substituted with one or more substituents independently chosen from R3, as described above. When Ar is a 6-membered heteroaryl, this group is
designated B D R3 , wherein variables are as described above.
Within certain embodiments, the group designated:
Figure imgf000024_0001
In certain such compounds, R3 represents from O to 2 substituents independently chosen from halogen, Ci-C4alkyl, Ci-C4hydroxyalkyl, Ci-C4alkoxy, C2-C4alkyl ether, and mono- or di-(Ci-C4alkyl)amino. One class of such compounds satisfies the Formula:
Figure imgf000024_0002
wherein each R9 is independently hydrogen or Ci-C4alkyl, or both Rg are taken together to form a spiro C3-C8cycloalkyl. In other such compounds, two substituents represented by R3 are taken together to form an optionally-substituted 5- to 7-membered fused carbocyclic or heterocyclic ring. Within one representative class of such
compounds, the group designated
Figure imgf000024_0003
IS , wherein
\ W) ^-^^ represents a 5- or 6-membered carbocycle or a 5- or 6-membered heterocycle, each of which is substituted with from O to 4 substituents independently chosen from halogen, hydroxy, amino, Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, and (4- to 7-membered heterocycle)C0-C4alkyl. Within certain compounds in which Ax is a 5-membered heteroaryl, this group is designated:
Figure imgf000025_0001
, wherein variables are as described above. In certain embodiments,
R3 represents 0 substituents or one substituent chosen from Ci-C4alkyl {e.g., methyl, ethyl or propyl), Ci-C4hydroxyalkyl and C2-C4alkyl ether. Certain such groups
Figure imgf000025_0002
S.
Representative Ar moieties include, for example:
Figure imgf000025_0003
wherein R3a is hydrogen, Ci-C4alkyl {e.g., methyl), Ci-C4hydroxyalkyl or C2-
C4alkyl ether. Certain such amide-substituted aryl piperidines satisfy the Formula:
Figure imgf000025_0004
Figure imgf000026_0001
Certain R4 moieties are indicated as "fused phenyl or 5- or 6-membered heterocyclic rings." In such groups, the ring represented by R4 is fused to the Z-containing ring, as in the following representative subformulas:
Figure imgf000026_0002
in which Yi, Y2, Y3 and Y4 are independently N, C, or CH and R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci- C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ci-C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl. In certain embodiments, Y] is N and Y2, Y3 and Y4 are each CH; in other embodiments, Y4 is N and Yi, Y2 and Y3 are each CH.
Other R4 moieties are "a phenyl or 5- or 6-membered heteroaryl substituent." In such groups, the ring represented by R4 is pendant, as in the following representative subformulas:
Figure imgf000027_0001
in which Yi, Y2, Y3, Y4 and Y5 are independently N, C, or CH; and R8 is as described above. In certain embodiments, Yi is N and Y2, Y3, Y4 and Y5 are each CH.
The variable "n" in Formulas I, III, IV, VII, VIII and IX (as well as the subformulas thereof) is generally 0, 1 or 2. Within certain compounds, n is 0 (i.e., the ring
designated
Figure imgf000027_0002
is 5-membered). Z is CH2 in certain such compounds. Within other compounds, n is 1, resulting in a 6-membered ring, or 2, resulting in a 7-membered ring. Formulas IHd-IIIh, IVd-IVh, VIIId-VIIIh and IXd-IXh illustrate certain amide- substituted aryl piperidines in which n is 0, 1 or 2, R5a is hydrogen, Ci-C6alkyl, C2- Cόalkenyl, C2-C6alkynyl or (C3-C8cycloalkyl)C0-C4alkyl and the remaining variables are as described above:
Figure imgf000027_0003
Formula IHd Formula IHe
Figure imgf000028_0001
Formula IHf Formula IHg
Figure imgf000028_0002
Formula HIh Formula FVd
Figure imgf000028_0003
Formula IVe Formula IVf
Figure imgf000028_0004
Formula IVg Formula IVh
Figure imgf000028_0005
Formula VIIId Formula VIIIe
Figure imgf000028_0006
Formula VIIIf Formula VIIIg
Figure imgf000029_0001
Formula VIIIh Formula IXd
Figure imgf000029_0002
Formula IXe Formula IXf
Figure imgf000029_0003
Formula IXg Formula IXh
Within compounds of Formulas II, V and VI, the group designated w is generally a heterocycle, as described above, that forms a spirocyclic system with the piperidine. Representative such spirocyclic systems include, for example:
Figure imgf000029_0004
in which Yi, Y2, Y3> Y4, Y5 and Rg is as described above. In certain embodiments, Yi is N and Y2, Y3> Y4 and Y5 (if present) are each CH; in other embodiments, Y4 is N and Yi, Y2, Y3 and Y5 (if present) are each CH.
The variables R5 (where present), R6 and R8 (where present) each represent O substituents within certain embodiments of the above Formulas.
Representative amide-substituted aryl piperidines provided herein include, but are not limited to, those specifically described in the Examples below. It will be apparent that the specific compounds recited herein are representative only, and are not intended to limit the scope of the present invention. Further, as noted above, all compounds of the present invention may be present as a free acid or base or as a pharmaceutically acceptable salt, hydrate or ester.
If desired, compounds provided herein may be evaluated for certain pharmacological properties including, but not limited to, oral bioavailability (preferred compounds are orally bioavailable to an extent allowing for therapeutically effective doses of less than 140 mg/kg, preferably less than 50 mg/kg, more preferably less than 30 mg/kg, even more preferably less than 10 mg/kg, still more preferably less than 1 mg/kg and most preferably less than 0.1 mg/kg), toxicity (a preferred compound is nontoxic when a therapeutically effective amount is administered to a subject), side effects (a preferred compound produces side effects comparable to placebo when a therapeutically effective amount of the compound is administered to a subject), serum protein binding and in vitro and in vivo half-life (a preferred compound exhibits an in vivo half-life allowing for Q.I.D. dosing, preferably T. LD. dosing, more preferably B. LD. dosing, and most preferably once- a-day dosing). In addition, differential penetration of the blood brain barrier may be desirable. Routine assays that are well known in the art may be used to assess these properties, and identify superior compounds for a particular use. For example, assays used to predict bioavailability include transport across human intestinal cell monolayers, including Caco-2 cell monolayers. Penetration of the blood brain barrier of a compound in humans may be predicted from the brain levels of the compound in laboratory animals given the compound (e.g., intravenously). Serum protein binding may be predicted from albumin binding assays. Compound half-life is inversely proportional to the frequency of dosage of a compound. In vitro half- lives of compounds may be predicted from assays of microsomal half-life as described herein.
As noted above, preferred compounds provided herein are nontoxic. In general, the term "nontoxic" as used herein shall be understood in a relative sense and is intended to refer to any substance that has been approved by the United States Food and Drug Administration ("FDA") for administration to mammals (preferably humans) or, in keeping with established criteria, is susceptible to approval by the FDA for administration to mammals (preferably humans). In addition, a highly preferred nontoxic compound generally satisfies one or more of the following criteria: (1) does not substantially inhibit cellular ATP production; (2) does not significantly prolong heart QT intervals; (3) does not cause substantial liver enlargement, or (4) does not cause substantial release of liver enzymes. As used herein, a compound that does not substantially inhibit cellular ATP production is a compound that satisfies the criteria set forth in Example 6, herein. In other words, cells treated as described in Example 6 with a 100 μM concentration of such a compound exhibit ATP levels that are at least 50% of the ATP levels detected in untreated cells. In more highly preferred embodiments, such cells exhibit ATP levels that are at least 80% of the ATP levels detected in untreated cells.
A compound that does not significantly prolong heart QT intervals is a compound that does not result in a statistically significant prolongation of heart QT intervals (as determined by electrocardiography) in guinea pigs, minipigs or dogs upon administration of a dose that yields a serum concentration equal to the EC50 or IC50 for the compound. In certain preferred embodiments, a dose of 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 40 or 50 mg/kg administered parenterally or orally does not result in a statistically significant prolongation of heart QT intervals. By "statistically significant" is meant results varying from control at the p<0.1 level or more preferably at the p<0.05 level of significance as measured using a standard parametric assay of statistical significance such as a student's T test.
A compound does not cause substantial liver enlargement if daily treatment of laboratory rodents (e.g., mice or rats) for 5-10 days with a dose that yields a serum concentration equal to the EC50 or IC50 for the compound results in an increase in liver to body weight ratio that is no more than 100% over matched controls. In more highly preferred embodiments, such doses do not cause liver enlargement of more than 75% or 50% over matched controls. If non-rodent mammals (e.g., dogs) are used, such doses should not result in an increase of liver to body weight ratio of more than 50%, preferably not more than 25%, and more preferably not more than 10% over matched untreated controls. Preferred doses within such assays include 0.01, 0.05. 0.1, 0.5, 1, 5, 10, 40 or 50 mg/kg administered parenterally or orally.
Similarly, a compound does not promote substantial release of liver enzymes if administration of twice the minimum dose that yields a serum concentration equal to the EC50 or IC50 for the compound does not elevate serum levels of ALT, LDH or AST in laboratory rodents by more than 100% over matched mock-treated controls, hi more highly preferred embodiments, such doses do not elevate such serum levels by more than 75% or 50% over matched controls. Alternatively, a compound does not promote substantial release of liver enzymes if, in an in vitro hepatocyte assay, concentrations (in culture media or other such solutions that are contacted and incubated with hepatocytes in vitro) that are equal to the EC50 or IC50 for the compound do not cause detectable release of any of such liver enzymes into culture medium above baseline levels seen in media from matched mock-treated control cells, hi more highly preferred embodiments, there is no detectable release of any of such liver enzymes into culture medium above baseline levels when such compound concentrations are five-fold, and preferably ten-fold the EC50 or IC50 for the compound.
In other embodiments, certain preferred compounds do not inhibit or induce microsomal cytochrome P450 enzyme activities, such as CYP 1A2 activity, CYP2A6 activity, CYP2C9 activity, CYP2C19 activity, CYP2D6 activity, CYP2E1 activity or CYP3 A4 activity at a concentration equal to the EC5O or IC50 for the compound. Certain preferred compounds are not clastogenic (e.g., as determined using a mouse erythrocyte precursor cell micronucleus assay, an Ames micronucleus assay, a spiral micronucleus assay or the like) at a concentration equal the EC50 or IC50 for the compound. In other embodiments, certain preferred compounds do not induce sister chromatid exchange (e.g., in Chinese hamster ovary cells) at such concentrations. For detection purposes, as discussed in more detail below, compounds provided herein may be isotopically-labeled or radiolabeled. For example, such compounds may have one or more atoms replaced by an atom of the same element having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be present in the compounds provided herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as 2H, 3H, 11C, 13C, 14C, 15N, 180, 170, 31P, 32P, 35S, 18F and 36Cl. In addition, substitution with heavy isotopes such as deuterium (i.e., 2H) can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances.
PREPARATION OF AMIDE-SUBSTITUTED ARYL PIPERIDINES
Compounds provided herein may generally be prepared using standard synthetic methods. In general, starting materials are commercially available from suppliers such as Sigma- Aldrich Corp. (St. Louis, MO), or may be synthesized from commercially available precursors using established protocols. By way of example, a synthetic route similar to that shown in any of the following Schemes may be used, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereon appreciated by those skilled in the art. It will be apparent that the reagents and synthetic transformations in the following Schemes and in the art can be readily modified to produce additional amide- substituted aryl piperidines. Each variable in the following Schemes refers to any group consistent with the description of the compounds provided herein.
When a protecting group is required, an optional deprotection step may be employed. Suitable protecting groups and methodology for protection and deprotection, such as those described in Protecting Groups in Organic Synthesis by T. Greene, are well known. Compounds and intermediates requiring protection/deprotection will be readily apparent.
Certain abbreviations used in the following Schemes and elsewhere herein include:
BINAP (rac)-2,2'-bis(diphenylphosphino)- 1 , 1 '-binaphthyl
BOC tert-butyloxycarbonyl
BOP benzotriazol- 1 -yl-oxy-tris(dimethylamino)phosphonium hexafluorophosphate
CDI N,N'-carbonyldiimidazole
DCE 1,1-dichloroethane
DCM dichloromethane
DMA N.N-dimethylacetamide
DMC 2-chloro- 1 ,3-dimethylimidazolidinium chloride
DMF dimethylformamide
DMSO dimethylsulfoxide
Eq. equivalent(s)
EtOAc ethyl acetate h hour(s)
1H NMR proton nuclear magnetic resonance
HOAc acetic acid
LC-MS liquid chromatography/mass spectrometry
LDA lithium diisopropyl amide
MCPBA m-chloroperoxybenzoic acid
MeOH methanol
MHz megahertz
M+H or M+l mass + 1 min minute(s)
MS mass spectrometry
NMP 1 -methyl-2-pyrrolidone δ chemical shift rt room temperature
PG protecting group
Ph phenyl
PTLC preparative thin layer chromatography t-BuOK potassium tert-butoxide
TEA triethylamine
TFA trifluoroacetic acid
THF tetrahydrofuran
Scheme 1
Figure imgf000034_0001
deprotection
Figure imgf000034_0002
Figure imgf000034_0003
Scheme 2
Figure imgf000034_0004
deprotection
Figure imgf000034_0005
Scheme 3
Figure imgf000035_0001
deprotection
Figure imgf000035_0002
Figure imgf000035_0003
deprotection
L=leaving group M= metal
Figure imgf000035_0004
Scheme 4
Figure imgf000035_0005
L = leaving group
R4NX = quaternary ammonium salt Scheme 5
base solvent
Figure imgf000036_0001
deprotection
Figure imgf000036_0003
Figure imgf000036_0002
Scheme 6
Figure imgf000036_0004
deprotection
Figure imgf000036_0006
Figure imgf000036_0005
Scheme 7
Figure imgf000037_0001
Scheme 8
Figure imgf000037_0002
Figure imgf000037_0003
L= leaving group
OR
Figure imgf000037_0004
Scheme 9
Figure imgf000038_0001
H2 / catalyst
Figure imgf000038_0002
Figure imgf000038_0003
Figure imgf000038_0004
Scheme 10
hydride / DMF
Figure imgf000038_0005
H2 / catalyst solvent
Figure imgf000038_0006
Figure imgf000038_0007
Scheme 11
Figure imgf000038_0008
In certain embodiments, a compound provided herein may contain one or more asymmetric carbon atoms, so that the compound can exist in different stereoisomeric forms. Such forms can be, for example, racemates or optically active forms. As noted above, all stereoisomers are encompassed by the present invention. Nonetheless, it may be desirable to obtain single enantiomers (i.e., optically active forms). Standard methods for preparing single enantiomers include asymmetric synthesis and resolution of racemates. Resolution of racemates can be accomplished, for example, by conventional methods such as crystallization in the presence of a resolving agent, or chromatography using, for example a chiral HPLC column.
Compounds may be radiolabeled by carrying out their synthesis using precursors comprising at least one atom that is a radioisotope. Each radioisotope is preferably carbon (e.g., 14C), hydrogen (e.g., 3H), sulfur (e.g., 35S) or iodine (e.g., 125I). Tritium labeled compounds may also be prepared catalytically via platinum-catalyzed exchange in tritiated acetic acid, acid-catalyzed exchange in tritiated trifluoroacetic acid, or heterogeneous- catalyzed exchange with tritium gas using the compound as substrate. In addition, certain precursors may be subjected to tritium-halogen exchange with tritium gas, tritium gas reduction of unsaturated bonds, or reduction using sodium borotritide, as appropriate. Preparation of radiolabeled compounds may be conveniently performed by a radioisotope supplier specializing in custom synthesis of radiolabeled probe compounds.
PHARMACEUTICAL COMPOSITIONS
The present invention also provides pharmaceutical compositions comprising one or more amide-substituted aryl piperidines provided herein, together with at least one physiologically acceptable carrier or excipient. Pharmaceutical compositions may comprise, for example, one or more of water, buffers (e.g., sodium bicarbonate, neutral buffered saline or phosphate buffered saline), ethanol, mineral oil, vegetable oil, dimethylsulfoxide, carbohydrates (e.g., glucose, mannose, sucrose, starch, mannitol or dextrans), proteins, adjuvants, polypeptides or amino acids such as glycine, antioxidants, chelating agents such as EDTA or glutathione and/or preservatives. In addition, other active ingredients may (but need not) be included in the pharmaceutical compositions provided herein.
Pharmaceutical compositions may be formulated for any appropriate manner of administration, including, for example, topical, oral (including, but not limited to, sublingual), nasal, rectal or parenteral administration. The term parenteral as used herein includes subcutaneous, intradermal, intravascular (e.g., intravenous), intramuscular, spinal, intracranial, intrathecal and intraperitoneal injection, as well as any similar injection or infusion technique. In certain embodiments, compositions suitable for oral use are preferred. Such compositions include, for example, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs. Within yet other embodiments, compositions of the present invention may be formulated as a lyophilizate.
Compositions intended for oral use may further comprise one or more components such as sweetening agents, flavoring agents, coloring agents and/or preserving agents in order to provide appealing and palatable preparations. Tablets contain the active ingredient in admixture with physiologically acceptable excipients that are suitable for the manufacture of tablets. Such excipients include, for example, inert diluents (e.g., calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate), granulating and disintegrating agents (e.g., corn starch or alginic acid), binding agents (e.g., starch, gelatin or acacia) and lubricating agents (e.g., magnesium stearate, stearic acid or talc). The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monosterate or glyceryl distearate may be employed.
Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent (e.g., calcium carbonate, calcium phosphate or kaolin), or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin or olive oil). Aqueous suspensions contain the active material(s) in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents (e.g., sodium carboxymethylcellulose, methylcellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia); and dispersing or wetting agents (e.g., naturally-occurring phosphatides such as lecithin, condensation products of an alkylene oxide with fatty acids such as polyoxyethylene stearate, condensation products of ethylene oxide with long chain aliphatic alcohols such as heptadecaethyleneoxycetanol, condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides such as polyethylene sorbitan monooleate).
Aqueous suspensions may also comprise one or more preservatives, such as ethyl or n- propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin. Oily suspensions may be formulated by suspending the active ingredient(s) in a vegetable oil (e.g., arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and/or flavoring agents may be added to provide palatable oral preparations. Such suspensions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, such as sweetening, flavoring and coloring agents, may also be present.
Pharmaceutical compositions may also be formulated as oil-in-water emulsions. The oily phase may be a vegetable oil (e.g., olive oil or arachis oil), a mineral oil (e.g., liquid paraffin) or a mixture thereof. Suitable emulsifying agents include naturally- occurring gums (e.g., gum acacia or gum tragacanth), naturally-occurring phosphatides (e.g., soy bean lecithin, and esters or partial esters derived from fatty acids and hexitol), anhydrides (e.g., sorbitan monoleate) and condensation products of partial esters derived from fatty acids and hexitol with ethylene oxide (e.g., polyoxyethylene sorbitan monoleate). An emulsion may also comprise one or more sweetening and/or flavoring agents.
Syrups and elixirs may be formulated with sweetening agents, such as glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also comprise one or more demulcents, preservatives, flavoring agents and/or coloring agents. Formulations for topical administration typically comprise a topical vehicle combined with active agent(s), with or without additional optional components. Suitable topical vehicles and additional components are well known in the art, and it will be apparent that the choice of a vehicle will depend on the particular physical form and mode of delivery. Topical vehicles include water; organic solvents such as alcohols (e.g., ethanol or isopropyl alcohol) or glycerin; glycols (e.g., butylene, isoprene or propylene glycol); aliphatic alcohols (e.g., lanolin); mixtures of water and organic solvents and mixtures of organic solvents such as alcohol and glycerin; lipid-based materials such as fatty acids, acyl glycerols (including oils, such as mineral oil, and fats of natural or synthetic origin), phosphoglycerides, sphingolipids and waxes; protein-based materials such as collagen and gelatin; silicone-based materials (both non- volatile and volatile); and hydrocarbon-based materials such as microsponges and polymer matrices. A composition may further include one or more components adapted to improve the stability or effectiveness of the applied formulation, such as stabilizing agents, suspending agents, emulsifying agents, viscosity adjusters, gelling agents, preservatives, antioxidants, skin penetration enhancers, moisturizers and sustained release materials. Examples of such components are described in Martindale— The Extra Pharmacopoeia (Pharmaceutical Press, London 1993) and Martin (ed.), Remington's Pharmaceutical Sciences. Formulations may comprise microcapsules, such as hydroxymethylcellulose or gelatin- microcapsules, liposomes, albumin microspheres, microemulsions, nanoparticles or nanocapsules.
A topical formulation may be prepared in a variety of physical forms including, for example, solids, pastes, creams, foams, lotions, gels, powders, aqueous liquids and emulsions. Typical modes of delivery for topical compositions include application using the fingers; application using a physical applicator such as a cloth, tissue, swab, stick or brush; spraying (including mist, aerosol or foam spraying); dropper application; sprinkling; soaking; and rinsing. Controlled release vehicles can also be used.
A pharmaceutical composition may be prepared as a sterile injectible aqueous or oleaginous suspension. The compound(s) provided herein, depending on the vehicle and concentration used, can either be suspended or dissolved in the vehicle. Such a composition may be formulated according to the known art using suitable dispersing, wetting and/or suspending agents such as those mentioned above. Among the acceptable vehicles and solvents that may be employed are water, 1,3-butanediol, Ringer's solution and isotonic sodium chloride solution, hi addition, sterile, fixed oils may be employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectible compositions, and adjuvants such as local anesthetics, preservatives and/or buffering agents can be dissolved in the vehicle.
Compounds may also be formulated as suppositories (e.g., for rectal administration). Such compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Suitable excipients include, for example, cocoa butter and polyethylene glycols. Pharmaceutical compositions may be formulated for release at a pre-determined rate. Instantaneous release may be achieved, for example, via sublingual administration (i.e., administration by mouth in such a way that the active ingredient(s) are rapidly absorbed via the blood vessels under the tongue rather than via the digestive tract). Controlled release formulations (i.e., formulations such as a capsule, tablet or coated tablet that slows and/or delays release of active ingredient(s) following administration) may be administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at a target site. In general, a controlled release formulation comprises a matrix and/or coating that delays disintegration and absorption in the gastrointestinal tract (or implantation site) and thereby provides a delayed action or a sustained action over a longer period. One type of controlled-release formulation is a sustained-release formulation, in which at least one active ingredient is continuously released over a period of time at a constant rate. Preferably, the therapeutic agent is released at such a rate that blood (e.g., plasma) concentrations are maintained within the therapeutic range, but below toxic levels, over a period of time that is at least 4 hours, preferably at least 8 hours, and more preferably at least 12 hours. Such formulations may generally be prepared using well known technology and administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at the desired target site. Carriers for use within such formulations are biocompatible, and may also be biodegradable; preferably the formulation provides a relatively constant level of modulator release. The amount of modulator contained within a sustained release formulation depends upon, for example, the site of implantation, the rate and expected duration of release and the nature of the condition to be treated or prevented.
Controlled release may be achieved by combining the active ingredient(s) with a matrix material that itself alters release rate and/or through the use of a controlled-release coating. The release rate can be varied using methods well known in the art, including (a) varying the thickness or composition of coating, (b) altering the amount or manner of addition of plasticizer in a coating, (c) including additional ingredients, such as release- modifying agents, (d) altering the composition, particle size or particle shape of the matrix, and (e) providing one or more passageways through the coating. The amount of modulator contained within a sustained release formulation depends upon, for example, the method of administration (e.g., the site of implantation), the rate and expected duration of release and the nature of the condition to be treated or prevented. The matrix material, which itself may or may not serve a controlled-release function, is generally any material that supports the active ingredient(s). For example, a time delay material such as glyceryl monosterate or glyceryl distearate may be employed. Active ingredient(s) may be combined with matrix material prior to formation of the dosage form (e.g., a tablet). Alternatively, or in addition, active ingredient(s) may be coated on the surface of a particle, granule, sphere, microsphere, bead or pellet that comprises the matrix material. Such coating may be achieved by conventional means, such as by dissolving the active ingredient(s) in water or other suitable solvent and spraying. Optionally, additional ingredients are added prior to coating (e.g., to assist binding of the active ingredient(s) to the matrix material or to color the solution). The matrix may then be coated with a barrier agent prior to application of controlled-release coating. Multiple coated matrix units may, if desired, be encapsulated to generate the final dosage form.
In certain embodiments, a controlled release is achieved through the use of a controlled release coating (i.e., a coating that permits release of active ingredient(s) at a controlled rate in aqueous medium). The controlled release coating should be a strong, continuous film that is smooth, capable of supporting pigments and other additives, nontoxic, inert and tack-free. Coatings that regulate release of the modulator include pH- independent coatings, pH-dependent coatings (which may be used to release modulator in the stomach) and enteric coatings (which allow the formulation to pass intact through the stomach and into the small intestine, where the coating dissolves and the contents are absorbed by the body). It will be apparent that multiple coatings may be employed (e.g., to allow release of a portion of the dose in the stomach and a portion further along the gastrointestinal tract). For example, a portion of active ingredient(s) may be coated over an enteric coating, and thereby released in the stomach, while the remainder of active ingredient(s) in the matrix core is protected by the enteric coating and released further down the GI tract. pH dependent coatings include, for example, shellac, cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropylmethylcellulose phthalate, methacrylic acid ester copolymers and zein. In certain embodiments, the coating is a hydrophobic material, preferably used in an amount effective to slow the hydration of the gelling agent following administration. Suitable hydrophobic materials include alkyl celluloses (e.g., ethylcellulose or carboxymethylcellulose), cellulose ethers, cellulose esters, acrylic polymers (e.g., poly(acrylic acid), poly(methacrylic acid), acrylic acid and methacrylic acid copolymers, methyl methacrylate copolymers, ethoxy ethyl methacrylates, cyanoethyl methacrylate, methacrylic acid alkamide copolymer, poly(methyl methacrylate), polyacrylamide, ammonio methacrylate copolymers, aminoalkyl methacrylate copolymer, poly(methacrylic acid anhydride) and glycidyl methacrylate copolymers) and mixtures of the foregoing. Representative aqueous dispersions of ethylcellulose include, for example,
AQUACOAT® (FMC Corp., Philadelphia, PA) and SURELEASE® (Colorcon, Inc., West Point, PA), both of which can be applied to the substrate according to the manufacturer's instructions. Representative acrylic polymers include, for example, the various EUDRAGIT® (Rohm America, Piscataway, NJ) polymers, which may be used singly or in combination depending on the desired release profile, according to the manufacturer's instructions.
The physical properties of coatings that comprise an aqueous dispersion of a hydrophobic material may be improved by the addition or one or more plasticizers. Suitable plasticizers for alkyl celluloses include, for example, dibutyl sebacate, diethyl phthalate, triethyl citrate, tributyl citrate and triacetin. Suitable plasticizers for acrylic polymers include, for example, citric acid esters such as triethyl citrate and tributyl citrate, dibutyl phthalate, polyethylene glycols, propylene glycol, diethyl phthalate, castor oil and triacetin.
Controlled-release coatings are generally applied using conventional techniques, such as by spraying in the form of an aqueous dispersion. If desired, the coating may comprise pores or channels to facilitate release of active ingredient. Pores and channels may be generated by well known methods, including the addition of organic or inorganic material that is dissolved, extracted or leached from the coating in the environment of use. Certain such pore-forming materials include hydrophilic polymers, such as hydroxyalkylcelluloses (e.g., hydroxypropylmethylcellulose), cellulose ethers, synthetic water-soluble polymers (e.g., polyvinylpyrrolidone, cross-linked polyvinylpyrrolidone and polyethylene oxide), water-soluble polydextrose, saccharides and polysaccharides and alkali metal salts. Alternatively, or in addition, a controlled release coating may include one or more orifices, which may be formed my methods such as those described in US Patent Nos. 3,845,770; 4,034,758; 4,077,407; 4,088,864; 4,783,337 and 5,071,607.
Controlled-release may also be achieved through the use of transdermal patches, using conventional technology (see, e.g., US Patent No. 4,668,232).
Further examples of controlled release formulations, and components thereof, may be found, for example, in US Patent Nos. 4,572,833; 4,587,117; 4,606,909; 4,610,870; 4,684,516; 4,777,049; 4,994,276; 4,996,058; 5,128,143; 5,202,128; 5,376,384; 5,384,133; 5,445,829; 5,510,119; 5,618,560; 5,643,604; 5,891,474; 5,958,456; 6,039,980; 6,143,353; 6,126,969; 6,156,342; 6,197,347; 6,387,394; 6,399,096; 6,437,000; 6,447,796; 6,475,493; 6,491,950; 6,524,615; 6,838,094; 6,905,709; 6,923,984; 6,923,988; and 6,911,217; each of which is hereby incorporated by reference for its teaching of the preparation of controlled release dosage forms.
In addition to or together with the above modes of administration, a compound provided herein may be conveniently added to food or drinking water (e.g., for administration to non-human animals including companion animals (such as dogs and cats) and livestock). Animal feed and drinking water compositions may be formulated so that the animal takes in an appropriate quantity of the composition along with its diet. It may also be convenient to present the composition as a premix for addition to feed or drinking water.
Amide-substituted aryl piperidines provided herein are generally administered in a therapeutically effective amount. Preferred systemic doses are no higher than 50 mg per kilogram of body weight per day (e.g., ranging from about 0.001 mg to about 50 mg per kilogram of body weight per day), with oral doses generally being about 5-20 fold higher than intravenous doses (e.g., ranging from 0.01 to 40 mg per kilogram of body weight per day). The amount of active ingredient that may be combined with the carrier materials to produce a single dosage unit will vary depending, for example, upon the patient being treated and the particular mode of administration. Dosage units will generally contain from about 10 μg to about 500 mg of an active ingredient. Optimal dosages may be established using routine testing, and procedures that are well known in the art.
Pharmaceutical compositions provided herein may, but need not, further comprise one or more additional pharmaceutical agents, such as an anti-inflammatory agent, analgesic, anti-migraine agent, serotonin agonist, anti-hypertensive and/or anticonvulsant.
Anti-inflammatory agents include, for example, non-steroidal anti-inflammatory drugs (NSAIDs), non-specific and cyclooxygenase-2 (COX-2) specific cyclooxgenase enzyme inhibitors, gold compounds, corticosteroids, methotrexate, leflunomide, cyclosporine A, EVI gold, minocycline, azathioprine, tumor necrosis factor (TNF) receptor antagonists, soluble TNF alpha receptor (etanercept), anti-TNF alpha antibodies (e.g., infliximab and adalimumab), anti-C5 antibodies, interleukin-1 (IL-I) receptor antagonists (e.g., anakinra or IL-I trap), IL-18 binding protein, CTLA4-Ig (e.g., abatacept), anti- human IL-6 receptor monoclonal antibody (e.g., tocilizumab), LFA-3-Ig fusion proteins (e.g., alefacept), LFA-I antagonists, anti-VLA4 monoantibody (e.g., natalizumab), anti- CD 1 Ia monoclonal antibody, anti-CD20 monoclonal antibody (e.g., rituximab), anti-IL-12 monoclonal antibody, anti-IL-15 monoclonal antibody, CDP 484, CDP 870, chemokine receptor antagonists, selective NOS inhibitors, p38 kinase inhibitors, integrin antagonists, angiogenesis inhibitors, and TMI-I dual inhibitors. Further anti-inflammatory agents include meloxicam, rofecoxib, celecoxib, etoricoxib, parecoxib, valdecoxib and tilicoxib.
NSAIDs include, but are not limited to, ibuprofen, flurbiprofen, naproxen or naproxen sodium, diclofenac, combinations of diclofenac sodium and misoprostol, sulindac, oxaprozin, difiunisal, piroxicam, indomethacin, etodolac, fenoprofen calcium, ketoprofen, sodium nabumetone, sulfasalazine, tolmetin sodium, and hydroxychloroquine. One class of NSAlDs consists of compounds that inhibit cyclooxygenase (COX) enzymes; such compounds include celecoxib and rofecoxib. NSAIDs further include salicylates such as acetylsalicylic acid or aspirin, sodium salicylate, choline and magnesium salicylates, and salsalate, as well as corticosteroids such as cortisone, dexamethasone, methylprednisolone, prednisolone, prednisolone sodium phosphate, and prednisone.
Certain analgesics for use in combination with CGRP receptor modulators provided herein are also anti-inflammatory agents, and are listed above. Other such medications include narcotic agents which typically act at one or more opioid receptor subtypes (e.g., μ, K and/or δ), preferably as agonists or partial agonists. Such agents include opiates, opiate derivatives and opioids, as well as pharmaceutically acceptable salts and hydrates thereof. Specific examples of narcotic analgesics include, within preferred embodiments, alfentanil, alphaprodine, anileridine, bezitramide, buprenorphine, butorphanol, codeine, diacetyldihydromorphine, diacetylmorphine, dihydrocodeine, diphenoxylate, ethylmorphine, fentanyl, heroin, hydrocodone, hydromorphone, isomethadone, levomethorphan, levorphane, levorphanol, meperidine, metazocine, methadone, methorphan, metopon, morphine, nalbuphine, opium extracts, opium fluid extracts, powdered opium, granulated opium, raw opium, tincture of opium, oxycodone, oxymorphone, paregoric, pentazocine, pethidine, phenazocine, piminodine, propoxyphene, racemethorphan, racemorphan, sulfentanil, thebaine and pharmaceutically acceptable salts and hydrates of the foregoing agents.
Other examples of narcotic analgesic agents include acetorphine, acetyldihydrocodeine, acetylmethadol, allylprodine, alphracetylmethadol, alphameprodine, alphamethadol, benzethidine, benzylmorphine, betacetylmethadol, betameprodine, betamethadol, betaprodine, clonitazene, codeine methylbromide, codeine-N-oxide, cyprenorphine, desomoφhine, dextromoramide, diampromide, diethylthiambutene, dihydromorphine, dimenoxadol, dimepheptanol, dimethylthiamubutene, dioxaphetyl butyrate, dipipanone, drotebanol, ethanol, ethylmethylthiambutene, etonitazene, etoφhine, etoxeridine, furethidine, hydromorphinol, hydroxypethidine, ketobemidone, levomoramide, levophenacylmorphan, methyldesoφhine, methyldihydromoφhine, moφheridine, moφhine methylpromide, moφhine methylsulfonate, moφhine-N-oxide, myrophin, naloxone, naltyhexone, nicocodeine, nicomoφhine, noracymethadol, norlevoφhanol, normethadone, normoφhine, noφipanone, pentazocaine, phenadoxone, phenampromide, phenomoφhan, phenoperidine, piritramide, pholcodine, proheptazoine, properidine, propiran, racemoramide, thebacon, trimeperidine and the pharmaceutically acceptable salts and hydrates thereof.
Further specific representative analgesic agents include, for example acetaminophen (paracetamol); aspirin and other NSAIDs described above; NR2B antagonists; bradykinin antagonists; anti-migraine agents; anticonvulsants such as oxcarbazepine and carbamazepine; antidepressants (such as TCAs, SSRIs, SNRIs, substance P antagonists, etc.); spinal blocks; pentazocine/naloxone; meperidine; levoφhanol; buprenoφhine; hydromoφhone; fentanyl; sufentanyl; oxycodone; oxycodone/acetaminophen, nalbuphine and oxymoφhone. Still further analgesic agents include CB2-receptor agonists, such as AM 1241, capsaicin receptor antagonists and compounds that bind to the α2δ subunit of voltage-gated calcium channels, such as gabapentin and pregabalin.
Representative anti-migraine agents for use in combination with a CGRP receptor modulator provided herein include caffeine; beta-adrenergic receptor antagonists such as timolol, propanolol, atenolol or nadolol; bradykinin- 1 receptor antagonists; adenosine Al receptor agonists; NMDA antagonists; NK-I receptor antagonists; opiate agonists; ergotamines; methysergide; calcium channel blockers such as flunarizine; selective serotonin uptake inhibitors such as fluoxetine, paroxetine, sertraline, duloxetine, escitalopram, or citalopram; 5-HTi agonists, such as sumatripan, naratriptan, zolmatriptan, eletriptan, almotriptan, frovatriptan, donitriptan and rizatriptan; inhibitors of nitric oxide synthesis; and cyclooxygenase inhibitors, as described above.
Additionally, or alternatively, additional pharmaceutical agents include decongestants such as phenylephrine, phenylpropanolamine, pseudoephedrine, oxymetazoline, epinephrine, naphazoline, xylometazoline, propylhexedrine, or levo- desoxy-ephedrine; sedating or non-sedating antihistamines; and/or asthma treatments such as beta-adrenergic receptor agonists; leukotriene D4 antagonists (e.g., montelukast or zafirlukast).
Pharmaceutical compositions may be packaged for treating conditions responsive to CGRP receptor modulation (e.g., treatment of pain, such as migraine or headache, or other disorder(s) recited herein). Packaged pharmaceutical preparations generally comprise a container holding a therapeutically effective amount of a pharmaceutical composition as described above and instructions (e.g., labeling) indicating that the composition is to be used for treating a condition responsive to CGRP receptor modulation in a patient (e.g., pain, such as migraine or headache, or other disorder as indicated herein). In certain embodiments, a packaged pharmaceutical preparation comprises one or more amide-substituted aryl piperidines provided herein and one or more additional agents in the same package, either in separate containers within the package or in the same container (i.e., as a mixture). Preferred mixtures are formulated for oral administration (e.g., as pills, capsules, tablets or the like). In certain embodiments, the package comprises a label bearing indicia indicating that the components are to be taken together for the treatment of pain.
METHODS OF USE
Within certain aspects, the present invention provides methods for treating a condition responsive to CGRP receptor modulation in a patient. The patient may be afflicted with such a condition, or may be free of symptoms but considered at risk for developing such a condition. A condition is "responsive to CGRP receptor modulation" if the condition or symptom(s) thereof are alleviated, attenuated, delayed or otherwise improved by modulation of CRP receptor activity. In general, such methods comprise administering to the patient a therapeutically effective amount of at least one amide- substituted aryl piperidine as provided herein.
Conditions responsive to CGRP receptor modulation include, for example pain (e.g., headache such as migraine), vasodilation (e.g., neurogenic vasodilation or in the treatment of shock, sepsis and hot flashes associated with menopause or hormone treatment), inflammation (e.g., neurogenic inflammation, inflammatory diseases of the joints such as arthritis, and airway inflammatory diseases such as asthma and chronic obstructive pulmonary disease), bronchial hyperreactivity, high blood pressure, hypotonia, tachycardia, diabetes (e.g., non-insulin-dependent diabetes mellitus), nausea, vomiting, diarrhea, loss of appetite, diabetic gastroparesis, polydipsis, thermal injury, cardiovascular diseases, morphine tolerance, skin diseases (e.g., dermatitis, psoriasis, thermal and radiation-induced skin damage such as sunburn), encephalitis, brain trauma, ischemia, epilepsy, neurodegenerative diseases, tinnitus, inflammatory bowel disease, irritable bowel syndrome, cystitis and allergic rhinitis. Headache that may be treated as described herein includes, for example, sinus, cluster (i.e., migranous neuralgia) and tension headaches, temporomandibular pain and maxillary sinus pain. Migraine headache and other migraine symptoms may also be treated; for example, migraine headaches may be prevented by administration of a compound provided herein as soon as a pre-migrainous aura is experienced by the patient. Other pain that may be treated using the CGRP receptor modulators provided herein may be chronic or acute and includes, but is not limited to, peripheral nerve- mediated pain (especially neuropathic pain, such as pain due to diabetes, postherpetic neuralgia, nerve injury, vulvodynia, root avulsions, painful traumatic neuropathy and painful polyneuropathy). Compounds provided herein may also be used in the treatment of, for example, visceral pain (e.g., pancreatitis, interstitial cystitis and renal colic), persistent hyperalgesia, inflammatory pain, repetitive motion pain, carpel tunnel syndrome, perioperative pain, algesia, oral neuropathic pain, toothache (dental pain), denture pain, ocular pain, postherpetic neuralgia, diabetic neuropathy, chemotherapy- induced neuropathy, reflex sympathetic dystrophy, trigeminal neuralgia, bone and joint pain (e.g., pain associated with osteoarthritis), rheumatoid arthritis, myofascial pain (e.g., muscular injury and fibromyalgia), Guillain-Barre syndrome, meralgia paresthetica, burning-mouth syndrome and/or pain associated with nerve and root damage, including as pain associated with peripheral nerve disorders (e.g., nerve entrapment and brachial plexus avulsions, amputation, peripheral neuropathies including bilateral peripheral neuropathy, tic douloureux, atypical facial pain, nerve root damage, and arachnoiditis). Additional neuropathic pain conditions include causalgia (reflex sympathetic dystrophy - RSD, secondary to injury of a peripheral nerve), neuritis (including, for example, sciatic neuritis, peripheral neuritis, polyneuritis, optic neuritis, postfebrile neuritis, migrating neuritis, segmental neuritis and Gombault's neuritis), neuronitis, neuralgias (e.g., those mentioned above, cervicobrachial neuralgia, cranial neuralgia, geniculate neuralgia, glossopharyngial neuralgia, migranous neuralgia, idiopathic neuralgia, intercostals neuralgia, mammary neuralgia, mandibular joint neuralgia, Morton's neuralgia, nasociliary neuralgia, occipital neuralgia, red neuralgia, Sluder's neuralgia, splenopalatine neuralgia, supraorbital neuralgia and vidian neuralgia), musculoskeletal pain, AIDS-related neuropathy, MS- related neuropathy, central nervous system pain (e.g., pain due to brain stem damage, sciatica, and ankylosing spondylitis), spinal pain, including spinal cord injury-related pain. Further pain conditions that may be treated include Charcot's pains, intestinal gas pains, ear pain, heart pain, muscle pain, eye pain, orofacial pain (e.g., odontalgia), abdominal pain, gynaecological pain (e.g., menstrual pain, dysmenorrhoea, pain associated with cystitis, labor pain, chronic pelvic pain, chronic prostitis and endometriosis), acute and chronic back pain (e.g., lower back pain), gout, scar pain, hemorrhoidal pain, dyspeptic pains, pain associated with angina, nerve root pain, "non-painful" neuropathies, complex regional pain syndrome, homotopic pain and heterotopic pain - including pain associated with carcinoma, often referred to as cancer pain (e.g., in patients with bone cancer), pain (and inflammation) associated with venom exposure (e.g., due to snake bite, spider bite, or insect sting) and trauma associated pain (e.g., post-surgical pain such as postmastectomy, postthoracotomy and stump pain; phantom limb pain, episiotomy pain, pain from cuts, musculoskeletal pain, bruises and broken bones, and burn pain, especially primary hyperalgesia associated therewith). Additional pain conditions that may be treated as described herein include pain associated with respiratory disorders as described above, autoimmune diseases, immunodeficiency disorders, hot flashes, inflammatory bowel disease, gastroesophageal reflux disease (GERD), irritable bowel syndrome and/or inflammatory bowel disease. It will be apparent that amide-substituted aryl piperidines provided herein may be administered alone or in combination with one or more additional agents that are suitable for treating the disorder of interest. Within such combination therapy, the compound(s) and additional agent(s) may be present in the same pharmaceutical composition, or may be administered separately in either order. Representative additional pharmaceutical agents for use in combination therapy include those indicated above administered by way of any means discussed above.
Treatment regimens may vary depending on the compound used and the particular condition to be treated. In general, a dosage regimen of 4 times daily or less is preferred, with 1 or 2 times daily particularly preferred. Dosage may be on an as-needed basis (e.g., the patient may self-administer upon the onset of symptoms). It will be understood, however, that the specific dose and treatment regimen for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex and diet of the patient, the time of administration, the route of administration, the rate of excretion, any drug combination and the severity of the particular disease undergoing therapy. Dosages are generally as described above; in general, the use of the minimum dose sufficient to provide effective therapy is preferred. Patients may generally be monitored for therapeutic effectiveness using medical or veterinary criteria suitable for the condition being treated or prevented. Dosages and methods of administration of any additional agent(s) can be found, for example, in the manufacturer's instructions or in the Physician 's Desk Reference. In certain embodiments, combination administration results in a reduction of the dosage of the additional agent required to produce a therapeutic effect {i.e., a decrease in the minimum therapeutically effective amount). Thus, preferably, the dosage of additional agent in a combination or combination treatment method of the invention is less than the maximum dose advised by the manufacturer for administration of the agent without combination with a CGRP receptor modulator provided herein. More preferably this dose is less than 3Zt, even more preferably less than Vi, and highly preferably less than 1A of the maximum dose, while most preferably the dose is less than 10% of the maximum dose advised by the manufacturer for administration of the agent(s) when administered without combination administration as described herein. It will be apparent that the dose of CGRP antagonist as provided herein needed to achieve the desired effect may similarly be reduced by the administration of the additional agent.
Within separate aspects, the present invention provides a variety of non- pharmaceutical in vitro and in vivo uses for the compounds provided herein. For example, such compounds may be labeled and used as probes for the detection and localization of CGRP receptor (in samples such as cell preparations or tissue sections, preparations or fractions thereof). In addition, compounds provided herein that comprise a suitable reactive group (such as an aryl carbonyl, nitro or azide group) may be used in photoaffϊnity labeling studies of receptor binding sites, m addition, compounds provided herein may be used as positive controls in assays for receptor activity, as standards for determining the ability of a candidate agent to bind to CGRP receptor, or as radiotracers for positron emission tomography (PET) imaging or for single photon emission computerized tomography (SPECT). Such methods can be used to characterize CGRP receptors in living subjects. For example, a compound may be labeled using any of a variety of well known techniques (e.g., radiolabeled with a radionuclide such as tritium, as described herein), and incubated with a sample for a suitable incubation time (e.g., determined by first assaying a time course of binding). Following incubation, unbound compound is removed (e.g., by washing), and bound compound detected using any method suitable for the label employed (e.g., autoradiography or scintillation counting for radiolabeled compounds; spectroscopic methods may be used to detect luminescent groups and fluorescent groups). As a control, a matched sample containing labeled compound and a greater (e.g., 10-fold greater) amount of unlabeled compound may be processed in the same manner. A greater amount of detectable label remaining in the test sample than in the control indicates the presence of CGRP receptor in the sample. Detection assays, including receptor autoradiography (receptor mapping) of CGRP receptor in cultured cells or tissue samples may be performed as described by Kuhar in sections 8.1.1 to 8.1.9 of Current Protocols in Pharmacology (1998) John Wiley & Sons, New York. Also provided herein are methods for detecting CGRP receptor agonist or antagonist activity of a test compound. Such methods generally employ cells that stably express CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2. Suitable cells include HEK293 cells that are transfected with recombinant DNAs that encode CRLR, RAMP-I and the rat olfactory cyclic nucleotide gated channel subunits 1 and 2 (GenBank # X55519 and #U12623, respectively). In certain embodiments, the
RAMP-I is human (e.g., GenBank #NM_005855) and the CRLR is human (e.g., GenBank #NM_005795). The resulting cells can take up extracellular calcium via the rat olfactory cyclic nucleotide gated channel in response to increased intracellular cyclic AMP that results from CGRP receptor activation. Such cells are then contacted with (a) a test compound; (b) CGRP receptor agonist
(e.g., CGRP); and (c) an indicator of intracellular calcium (e.g., a membrane permeable calcium sensitivity dye such as Fluo-4 AM, which produces a fluorescent signal when bound to Ca++). Following sufficient contact to allow the indicator to enter the cells, a response indicative of intracellular calcium level in the contacted cells is measured. If the indicator produces a fluorescent response, typically the maximum response observed over a period of time (e.g., 4 minutes) is determined. This response is compared to a response detected in control cells that are treated identically to those described above, except the absence of test compound. Compounds with antagonist activity reduce fluorescent signal detected in the presence of agonist and indicator. The following Examples are offered by way of illustration and not by way of limitation. Unless otherwise specified all reagents and solvent are of standard commercial grade and are used without further purification. Using routine modifications, the starting materials may be varied and additional steps employed to produce other compounds provided herein. EXAMPLES
Mass spectroscopy in the following Examples is Electrospray MS, obtained in positive ion mode using a Waters ZMD II Mass Spectrometer (Waters Corp.; Milford, MA), equipped with a Waters 600 pump (Waters Corp.), Waters 996 photodiode array detector (Waters Corp.), and a Gilson 215 autosampler (Gilson, Inc.; Middleton, WI). MassLynx™ version 4.0 software with OpenLynx processing is used for data collection and analysis (Waters Corp., Milford MA). MS conditions are: capillary voltage = 3.5 kV; cone voltage = 30 V, desolvation and source temperature = 250 0C and 100 0C, respectively; mass range = 100-750 with a scan time of 0.75 seconds and an interscan delay of 0.15 minutes. LCMS conditions are as follows:
Column 4.6x50mm, Chromolith SpeedRod (Merck KGaA);
UV 5 spectra/sec
Extracted wavelengths 220 and 254nm
Flow rate 6.0 mL/min
Injection Volume l-10μl;
Mobile phase A 95% Water, 5% Methanol with 0.05% Formic acid
Mobile phase B 95% Methanol, 5% Water with 0.025% Formic acid
Gradient:
Time(min) %B
0 5
0.01 5
1.0 100
2 100
2.1 5
EXAMPLE 1. SYNTHESIS OF REPRESENTATIVE INTERMEDIATES
This Example illustrates the preparation of representative intermediates useful in the synthesis of certain amide-substituted aryl piperidines.
1. N-(I -Benzylpiperidin-4-yl)-2-(2-nitrophenyl)acetamide
Figure imgf000054_0001
A solution of 2-nitrophenylacetic acid (60 g, 331 mmol) in 150 ml of THF is added to a suspension of CDI (59 g, 364 mmol) in 150 ml of THF, during which time CO2 (g) evolution occurs. After the addition, the brown reaction mixture is stirred for 30 min. at rt. A solution of 4-amino-l-benzylpiperidine (67.5 ml, 331 mmol) in 30 ml of THF is then added dropwise (the amine addition is mildly exothermic). The resulting reaction mixture is stirred for 2 h at rt. A little THF (about 150 ml) is removed in vacuo, and water (600 ml) is added. The resulting suspension is stirred for 1 h at 0 °C. The solid is filtered, washed with Et2O, and dried in vacuo to give the title compound as a white solid. MS: 354.16 (M+H).
2. 1 -Benzyl-N-r2-(2-nitrophenyl)ethyl1piperidin-4-amine bis-hvdrochloride
Figure imgf000055_0001
A solution of N-(l-benzylpiperidin-4-yl)-2-(2-nitrophenyl)acetamide (42 g, 119 mmol) in 350 ml of THF is heated to 60 0C, and chlorotrimethylsilane (54 ml, 428.4 mmol) is added. A white solid precipitates from solution. Lithium borohydride (2.0M in THF, 143 ml, 285.6 mmol) is then added dropwise over 15-20 min., during which time gas evolution is observed and the solids in the mixture slowly dissolve. After the addition, the reaction mixture is stirred for 4 h at 60 °C. After cooling to rt, 35 ml of MeOH is added, followed by 65 ml of 30% aq. HCl. The resulting mixture is refluxed for 2.5 h, and then once again cooled to rt. The pH is adjusted to 9-10 with a 50% aq. NaOH solution, the mixture is transferred to a separating funnel and the milky white aqueous layer is separated. To the organic phase is added 30 ml of 30% aq. HCl, and the resulting mixture is heated to reflux for 1 h. The mixture is then cooled to 0 °C and kept at that temperature overnight. The solid is then filtered, washed with Et2O, and dried in vacuo to give the title compound as a white solid. MS: 340.20 (M+H).
3. 3-(l-Benzylpiperidin-4-yl)-l,3A5-tetrahvdro-2H-l,3-benzodiazepin-2-one
Figure imgf000055_0002
To a suspension of l-benzyl-N-[2-(2-nitrophenyl)ethyl]piperidin-4-amine bis- hydrochloride (30 g, 73 mmol) in 150 ml of toluene and 150 ml of water is added 18 g of a 50% aq. NaOH solution. The resulting mixture is stirred for 1 h at 50 0C. The layers are separated, and the organic phase is washed with water and concentrated in vacuo. The brown oily residue is dissolved in 150 ml of MeOH and treated with Raney Ni (2.7 g, slurry in water), and the resulting reaction mixture is hydrogenated under 30-40 psi of H2(g) for 2.5 h. The mixture is filtered through Celite, and the Celite is washed well with MeOH. The filtrate is then concentrated to give a dark brown oil, which is dissolved in 60 ml of THF and added dropwise to a suspension of CDI (22 g, 128 mmol) in 60 ml of THF. After the addition, the reaction is stirred for 1 h at rt. Half of the THF is removed in vacuo, and the remaining solution is cooled to 0 0C in an ice bath. Water (100 ml) is added, and the resulting precipitate is filtered, washed with Et2O, and dried in vacuo to give the title compound as an off-white solid. MS: 336.19 (M+H).
4. 3-Piperidin-4-yl-l,3A5-tetrahvdro-2H-l,3-benzodiazepin-2-one trifluoroacetate
Figure imgf000056_0001
A mixture of 3-(l-benzylpiperidin-4-yl)-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin- 2-one (22.5 g, 67.1 mmol) and 10% Pd/C (2.2 g) in 500 ml of MeOH is hydrogenated at rt under 40 psi of H2 (g) for 3 days. The catalyst is removed by filtration through Celite, and the Celite is washed well with MeOH. The filtrate is concentrated in vacuo to give a brown solid, which is taken up in CH2Cl2, cooled to 0 °C, and treated with TFA (12-13 ml). The resulting mixture is concentrated, and the residue is triturated with Et2O and filtered to give the title compound as an off-white solid. MS: 245.15 (M+H). 5. 4-rBenzyl(methyl)aminol-3,3-dimethylbutan-2-one
Figure imgf000056_0002
A mixture of isopropyl methyl ketone (4.1 ml, 38.1 mmol), N-methylbenzylamine hydrochloride (6.0 g, 38.1 mmol), and formaldehyde (37% in water, 10.2 ml, 137.2 mmol) is heated to 100 0C and stirred vigorously for 15 h in a sealed tube. After cooling to rt, the homogenous solution is extracted with Et2O (20 ml) to remove any unreacted ketone. The aqueous layer is basified to pH=10-l 1 with 30% NaOH and extracted with Et2O (3 x 30 ml). The combined ethereal extracts are dried (Na2SO4), filtered, and evaporated to give a yellow/brown oil. Purification by silica gel column chromatography (gradient from Hex to 10% EtO Ac/Hex) affords the title compound as a clear oil. 1H NMR (400 MHz, CDCl3) δ 7.30 (5H, m), 3.51 (2H, s), 2.61 (2H, s), 2.16 (3H, s), 2.13 (3H, s), 1.15 (6H, s). 6. N-Benzyl-N,2-dimethyl-2-(lH-pyrazol-5-yl)propan-l -amine
Figure imgf000057_0001
A mixture of 4-[benzyl(methyl)amino]-3,3-dimethylbutan-2-one (1.65 g, 7.52 mmol) and ethyl formate (0.6 ml, 7.52 mmol) is added slowly to a slurry of freshly prepared sodium methoxide (419 mg, 7.75 mmol) in 10 ml of anhydrous Et2O. After the addition, an additional 0.16 ml (2.0 mmol) of ethyl formate is added. The resulting reaction mixture is stirred for 1 h at RT, at which time a solution of hydrazine monohydrate (0.38 ml, 7.75 mmol) and 0.7 ml of cone. HCl in 2.5 ml of water is added. After stirring for 15 h at rt, the layers are separated, and the ethereal layer is washed twice with brine (10 ml each), dried (Na2SO4), filtered, and evaporated in vacuo to give a light yellow oil. Purification by silica gel column chromatography (gradient from Hex to 50% EtO Ac/Hex) affords the title compound as a viscous, pale yellow oil. MS: 245.29 (M+H).
7. fert-Butyl 5-{2-rbenzyl(methyl')aminol-l,l-dimethylethvU-lH-pyrazole-l- carboxvlate
Figure imgf000057_0002
A solution of di-tert-butyldicarbonate (567 mg, 2.60 mmol), 4- dimethylaminopyridine (21 mg, 0.173 mmol), and TEA (3.5 ml) in 7 ml OfCH2Cl2 is added to a solution of N-benzyl-N,2-dimethyl-2-(lH-pyrazol-5-yl)propan-l -amine (420 mg, 1.73 mmol) in 3 ml Of CH2Cl2. After stirring for 14 h at rt, water (10-15 ml) is added, and the layers are separated. The CH2Cl2 layer is dried (Na2SO4), filtered, and evaporated in vacuo to give a brown oil. Purification by silica gel column chromatography (20%
EtO Ac/Hex) affords the title compound as a viscous, light brown oil. MS: 44.23 (M+H). 8. fert-Butyl 5 -I" 1 , 1 -dimethyl-2-(methylamino)ethyll - 1 H-p yrazole- 1 -carboxvlate
NH
>*O \
O
A mixture of tert-butyl 5-{2-[benzyl(methyl)amino]-l,l-dimethylethyl}-lH- pyrazole-1-carboxylate (400 mg, 1.16 mmol) and 10% Pd/C (100 mg) in 30 ml of EtOH is hydrogenated under 30-40 psi of H2 (g) on a Parr shaker for 15 h. The reaction is filtered through Celite, and the Celite is washed well with EtOH. The filtrate is concentrated in vacuo to give the title compound as a light brown oil. MS: 254.17 (M+H).
9. tert-Butyl 5-(2-ilY2-chloro-4-methyl-L3-thiazol-5- yl)carbonyl1(methyl)amino 1-1,1 -dimethylethyl)- 1 H-p yrazole- 1 -carboxylate
Figure imgf000058_0001
2-Chloro-l,3-dimethylimidazolinium chloride (272 mg, 1.61 mmol) is added portionwise to a stirred mixture of 2-chloro-4-methylthiazole-5-carboxylic acid (190 mg, 1.07 mmol), /ert-butyl 5-[l,l-dimethyl-2-(methylamino)ethyl]-lH-pyrazole-l-carboxylate (270 mg, 1.07 mmol), and TEA (0.45 ml, 3.21 mmol) in 10 ml OfCH2Cl2 at 0 °C. After 10-15 minutes, water (10 ml) is added, and the layers are separated. The CH2Cl2 layer is dried (Na2SO4), filtered, and evaporated in vacuo to give a yellow/brown oil. Purification by silica gel column chromatography (gradient from CH2Cl2 to 10% EtOAc/CH2Cl2) affords the title compound as pale yellow oil. MS: 313.13 [M+H (-Boc)]. 10. Methyl 4-chloropicolinate
Figure imgf000058_0002
A mixture of picolinic acid (15 g, 122 mmol) sodium bromide (1.88 g, 18.3 mmol), and thionyl chloride (45 ml) is heated to reflux and stirred for 20 h. After cooling to rt, the excess thionyl chloride is removed in vacuo. The resulting residue is taken up in toluene (100 ml) and cooled to 0 °C. MeOH (7 ml) is added dropwise, after which time the reaction is warmed to rt and stirred an additional 1.5 h. The precipitated solid is collected by filtration, washed with toluene, and partitioned between EtOAc (200 ml) and water (100 ml). The mixture is neutralized to pH=7-8 with solid NaHCO3. The EtOAc layer is dried (Na2SO4), filtered, and evaporated in vacuo to give the title compound as a light brown solid. MS: 171.94 (M+H). 11. 4-Chloropicolinic acid
Figure imgf000059_0001
A 50% aq. NaOH solution (8 ml) is added to a solution of methyl 4- chloropicolinate (3.6 g, 21 mmol) in 20 ml of MeOH at 0 °C. After the addition, the ice bath is removed, and the suspension is stirred at rt for 15 minutes. The MeOH is removed in vacuo, and the residue is diluted with water (20 ml) and acidified to pH=2 with 6N HCl. The mixture is then filtered, and the solid is dried in vacuo to give the title compound as an off-white solid. MS: 158.10 (M+H).
12. 5-r(4-Chloropyridin-2-yl)carbonyll-7,7-dimethyl-4.5.6.7-tetrahvdro-lH- pyrazolo [4,3 -dp yri dine
Figure imgf000059_0002
A mixture of 4-chloropicolinic acid (200 mg, 1.27 mmol) and thionyl chloride (1.0 ml) is heated to reflux for 1 h. The excess thionyl chloride is removed in vacuo to give the acid chloride as a brown solid, which is dissolved in CH2Cl2 (8.0 ml)and added to a mixture of 7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis-hydrochloride (285 mg, 1.27 mmol) and TEA (0.71 ml, 5.08 mmol) in 7.0 ml OfCH2Cl2 cooled to 0 °C. After 20 min. at 0 °C, water (5 ml) is added, and the reaction is warmed to rt. The layers are separated, and the aqueous layer is extracted once more with CH2Cl2 (10 ml). The combined organic extracts are dried (Na2SO4), filtered, and evaporated to give a yellow/green oil. Purification by silica gel column chromatography (gradient from 50% EtOAc/CH2Cl2 to 1% MeOH/EtOAc) affords the title compound as a white solid. MS: 291.18 (M+H).
13. 5-r(2-Chloro-4-methyl-l,3-thiazol-5-vncarbonyll-7.7-dimethyl-4.5.6.7- tetrahvdro-lH-pyrazolo[4,3-clpyridine
Figure imgf000059_0003
2-Chloro-l,3-dimethylimidazolinium chloride (315 mg, 1.86 mmol) is added portionwise to a stirred mixture of 2-chloro-4-methylthiazole-5-carboxylic acid (220 mg, 1.24 mmol), 7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis- hydrochloride (278 mg, 1.24 mmol) and TEA (0.69 ml, 4.96 mmol) in 10 ml OfCH2Cl2 at 0 °C. After 10-15 minutes, water (10 ml) is added, and the layers are separated. The CH2Cl2 layer is dried (Na2SO4), filtered, and evaporated in vacuo to give a yellow oil. Purification by silica gel column chromatography (gradient from 50% EtO Ac/Hex to EtOAc) affords the title compound as a clear oil. MS: 311.07 (M+H ).
14. 5-(2-Chloro-6-methylisonicotinoylV7.7-dimethyl-4.5.6.7-tetrahydro-lH- pyrazolor4,3-c1pγridine
Figure imgf000060_0001
A mixture of 2-chloro-6-methylpyridine-4-carboxylic acid (5.0 g, 29.1 mmol) and thionyl chloride (20 ml) is heated to reflux and stirred for 5 hours. The volatiles are removed in vacuo to give the acid chloride as a light purple oil, which solidifies while standing in the freezer. A solution of this acid chloride (1.3 g, 6.7 mmol) in 10 ml of CH2Cl2 is then added dropwise to a mixture of 7,7-dimethyl-4,5,6,7-tetrahydro-lH- pyrazolo[4,3-c]pyridine bis-hydrochloride (1.5 g, 6.7 mmol) and TEA (3.7 ml, 26.8 mmol) in 15 ml OfCH2Cl2 cooled to 0 °C. After stirring for 15 min. at 0 °C, water (15 ml) is added, and the reaction is warmed to rt. The layers are separated, and the aqueous layer is extracted once more with CH2Cl2 (20 ml). The combined organic extracts are dried
(Na2SO4), filtered, and evaporated to give a yellow oily solid. Purification by silica gel column chromatography (gradient from 50% EtOAc/CH2Cl2 to EtOAc) affords the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.39 (IH, s), 7.25 (IH, s), 7.14 (IH, s), 7.08 (IH, s), 4.82 (IH, s), 4.40 (IH, s), 3.78 (IH, s), 3.33 (IH, s), 2.57 (3H, d, J 4.4), 1.39 (3H, s), 1.25 (3H, s). MS: 305.17 (M+H).
15. Methyl 2-chloro-6-methylpyridine-4-carboxvlate-N-oxide
Figure imgf000061_0001
A solution of MCPBA (6.6 g, 29.6 mmol) in 50 ml OfCH2Cl2 is added dropwise to a solution of methyl 2-chloro-6-methylpyridine-4-carboxylate (5.0 g, 26.9 mmol) in 30 ml OfCH2Cl2 at RT. The reaction mixture is stirred for 2 days, and then it is evaporated to dryness. Sat. NaHCO3 (20 ml) and EtOAc (30 ml) are added. The layers are separated, and the aqueous layer is extracted twice more with EtOAc (30 ml each). The combined organic extracts are dried (Na2SO4), filtered, and evaporated in vacuo to give an off-white solid. Purification by silica gel column chromatography (gradient from Hex to EtOAc) affords the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.0 (IH, s), 7.80 (IH, s), 3.94 (3H, s), 2.57 (3H, s).
16. Methyl 2-chloro-6-(chloromethvl)isonicotinate
Figure imgf000061_0002
A mixture of methyl 2-chloro-6-methylpyridine-4-carboxylate-N-oxide (3.4 g, 16.9 mmol) and phosphorous oxychloride (5.0 ml) is heated to 100 °C and stirred for 48 h. The excess POCl3 is removed in vacuo, and the residue is dissolved in CH2Cl2 and washed with sat. NaHCO3. The CH2Cl2 layer is dried (Na2SO4), filtered, and evaporated in vacuo to give a dark brown oil. Purification by silica gel column chromatography (gradient from Hex to 20% EtO Ac/Hex) affords the title compound as yellow/brown oil. MS: 220.01 (M+H). 17. 2-Chloro-6-(methoxymethyl)isonicotinic acid
Figure imgf000061_0003
A sodium methoxide solution (25 wt. % in MeOH, 5.2 ml, 22.8 mmol) is added dropwise to a solution of methyl 2-chloro-6-(chloromethyl)isonicotinate (2.5 g, 11.4 mmol) in 20 ml of THF and 2 ml of water at rt. After 3.5 h, the reaction is diluted with water and extracted with Et2O. The aqueous layer is cooled to 0 0C and acidified to pH=2 with cone. HCl. The cloudy mixture is extracted with CH2Cl2 (2 x 30 ml), and the combined organic extracts are dried (Na2SO4), filtered, and evaporated in vacuo to give the title compound as a brown solid. MS: 202.07 (M+H).
18. 5-r2-Chloro-6-(methoxymethyl)isonicotinoyll-7,7-dimethyl-4,5,6,7- tetrahvdro-lH-pyrazolor4,3-clpyridine
Figure imgf000062_0001
2-Chloro-l,3-dimethylimidazolinium chloride (2.5 g, 14.88 mmol) is added portionwise to a stirred mixture of 2-chloro-6-(methoxymethyl)-isonicotinic acid (2.0 g, 9.92 mmol), 7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis- hydrochloride (2.2 g, 9.92 mmol) and TEA (5.5 ml, 39.68 mmol) in 30 ml Of CH2Cl2 at 0 °C. After 20 min, water (20 ml) is added, and the layers are separated. The aqueous layer is extracted once more with 20 ml OfCH2Cl2. The combined CH2Cl2 extracts are dried (Na2SO4), filtered, and evaporated to give a yellow/brown oil. Purification by silica gel column chromatography (gradient from 40% EtOAc/ CH2Cl2 to EtOAc) affords the title compound as a viscous, yellow oil. MS: 335.16 (M+H ).
19. 5-(2.6-Dichloroisonicotinoyl)-7.7-dimethyl-4,5,6J-tetrahvdro-lH- p yrazolo [4,3 -dp yridine
C
Figure imgf000062_0002
2-Chloro-l,3-dimethylimidazolinium chloride (254 mg, 1.5 mmol) is added portionwise to a stirred mixture of 2,6-dichloroisonicotinic acid (192 mg, 1.0 mmol), 7,7- dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis-hydrochloride (224 mg, 1.0 mmol) and TEA (0.56 ml, 4.0 mmol) in 10 ml OfCH2Cl2 at 0 0C. After 10 minutes, water (10 ml) is added, and the layers are separated. The aqueous layer is extracted once more with 10 ml Of CH2Cl2. The combined CH2Cl2 extracts are dried (Na2SO4), filtered, and evaporated to give a brown oil. Purification by silica gel column chromatography (gradient from 50% EtOAc/ CH2Cl2 to EtOAc) affords the title compound as a pale brown solid. MS: 325.11 (M+H).
20. 5-r2-Chloro-6-(4-pyridin-4-ylpiperidin-l-yl)isonicotinoyll-7,7-dimethyl- 4,5,6J-tetrahydro-lH-pyrazolor4,3-c1pyridine
Figure imgf000063_0001
A mixture of 5-(2,6-dichloroisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro-lH- pyrazolo[4,3-c]pyridine (220 mg, 0.68 mmol), 4-piperidin-4-ylpyridine (110 mg, 0.68 mmol), and potassium carbonate (188 mg, 1.36 mmol) in 2.0 ml of dioxane is heated to 110 0C and stirred for 15 h in a sealed tube. The dioxane is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50%
EtOAc/CH2Cl2 to 5% MeOH/ CH2Cl2) to give the title compound as a white solid. MS: 451.20 (M+H ).
21. 5-(2-Chloroisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahvdro-lH-pyrazolor4,3- clpyridine
Figure imgf000063_0002
2-Chloro-l,3-dimethylimidazolinium chloride (323 mg, 1.91 mmol) is added portionwise to a stirred mixture of 2-chloroisonicotinic acid (200 mg, 1.27 mmol), 7,7- dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine bis-hydrochloride (285 mg, 1.27 mmol) and TEA (0.71 ml, 5.08 mmol) in 15 ml OfCH2Cl2 at 0 °C. After 10 minutes, water (15 ml) is added, and the layers are separated. The aqueous layer is extracted once more with 15 ml Of CH2Cl2. The combined CH2Cl2 extracts are dried (Na2SO4), filtered, and evaporated to give a brown oil. Purification by silica gel column chromatography (gradient from 50% EtOAc/ CH2Cl2 to EtOAc) affords the title compound as a pale brown solid. MS: 291.17 (M+H ). 22. Ethyl 4-methyl-2-(4-(2-oxo-l .2.4.5-tetrahvdrobenzordl [ 1 ,31diazepin-3- vπpiperidin-l-yl)thiazole-5-carboxylate
Figure imgf000064_0001
3-Piperidin-4-yl-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one trifluoroacetate (3 g, 8.35 mmol), ethyl 2-bromo-4-methyl-l,3-thiazole-5-carboxylate (2.1 g, 8.35 mmol) and potassium carbonate (3.46 g, 25.1 mmol) in NMP (20 mL) is heated at 150 0C for 2 hours. The mixture is then cooled to rt, diluted with EtOAc, washed with water and brine, dried over Na2SO4, and concentrated under reduced pressure. Purification by silica gel chromatography (hexane / EtOAc = 1/1) affords the title compound as a white solid. 1H- NMR (400 MHz, CDCl3) δ: 7.11 (m, IH), 7.05 (d, IH), 6.91 (m, IH), 6.70 (d, IH), 6.42 (s, IH), 4.51 (m, IH), 4.26 (q, 2H), 4.13-4.20 (m, 2H), 3.45-3.47 (m, 2H), 3.13-3.20 (m, 2H), 2.96-3.00 (m, 2H), 2.55 (s, 3H), 1.74-1.89 (m, 4H), 1.33 (t, 3H). MS: 415.25 (M+H).
23. 4-Methyl-2-(4-(2-oxo- 1 ,2,4,5-tetrahvdrobenzordir 1 ,31diazepin-3-yl)piperidin- l-yl)thiazole-5-carboxylic acid
Figure imgf000064_0002
Ethyl4-methyl-2-(4-(2-oxo-l,2,4,5-tetrahydrobenzo[d][l,3]diazepin-3-yl)piperidin- l-yl)thiazole-5-carboxylate (1.56 g, 3.77 mmol) is treated with IN aq. NaOH (18.8 mL) and EtOH (18.8 mL) at 70 °C for 4 hours. The reaction is cooled to rt, and EtOH is removed under reduced pressure to give a white suspension. The pH of this suspension is adjusted to 5. The white precipitates are collected via filtration, and dried under vacuum overnight to afford the title compound. 1H-NMR (400 MHz, CDCl3) δ: 7.45 (s, IH), 7.11 (m, IH), 7.05 (d, IH), 6.91(m, IH), 6.82 (d, IH), 4.55 (m, IH), 4.21 (d, 2H), 3.47 (bs. 2H), 3.16-3.22 (m, 2H), 3.00 (bs, 2H), 2.58 (s, 3H), 1.78-1.90 (m, 5H). MS: 387.18 (M+l). 24. 1 -(2-Chloroisonicotinoyl)-3,3-dimethylpiperidin-4-one
Figure imgf000064_0003
To a solution of 3,3-dimethylpiperidin-4-one HCl salt (7.3g, 45 mmol) in DMA (100 mL) is added TEA (24.9 mL, 180 mmol). After 5 min of stirring, 2-chloro- isonicotinic acid (7.80 g, 49.5 mmol) and BOP (29.8 g, 67.5 mmol) are added. The mixture is stirred at 4O0C overnight. The reaction is cooled to rt, diluted with EtOAc, washed with 50% saturated Na2CO3, water and brine, dried over Na2SO4, and concentrated under reduced pressure. The residue is purified by silica gel chromatography (hexane : EtOAc = 1:1) to afford the title compound as a yellow solid. 1H-NMR (400 MHz, CDCl3) δ: 8.51 (d, IH), 7.38 (s, IH), 7.25 (d, IH), 4.03 (t, IH), 3.77 (s, IH), 3.64 (t, IH), 3.36 (s, IH), 2.65 (t, IH), 2.51 (t, IH), 1.23 (s, 3H), 1.05 (s, 3H). MS: 267.08 (M+H). 25. tert-Butyl 4-r(2-ethoxy-2-oxoethyl)amino1piperidine- 1 -carboxylate
Figure imgf000065_0001
To a solution of l-Boc-4-piperidone (25 g, 125 mmol) and glycine ethyl ester hydrochloride (17.5 g, 125 mmol) in 625 ml of methanol is added sodium cyanoborohydride (9.45 g, 150 mmol) in small portions. Evolution of gas is observed. After 16 h, the mixture is quenched with sat. NH4Cl, concentrated, and partitioned between CH2Cl2 and sat. NaHCO3. The organic layer is washed with brine, dried (Na2SO4), filtered, and concentrated. Purification by silica column chromatography [gradient from CH2Cl2 to 95% CH2Cl2/5% (10% NH4OHMeOH)] affords the title compound as clear, viscous oil.
26. tert-Butyl 4-(2,4-dioxoimidazolidin-l-yl)piperidine-l -carboxylate
Figure imgf000065_0002
Potassium cyanate (8.7 g, 107 mmol) is added to a solution of tert-butyl 4-[(2- ethoxy-2-oxoethyl)amino]piperidine-l-carboxylate (30.5 g, 107 mmol) in water (600 ml). The pH of the reaction mixture is adjusted to 4-5 with acetic acid, and the resulting reaction mixture is heated to 40 0C for 16 h. After cooling to rt, the mixture is filtered and the solid dried in vacuo to give the title compound (with some inorganic salts) as a white solid which is used without further purification.
27. l-Piperidin-4-ylimidazolidine-2,4-dione trifluoroacetate
Figure imgf000066_0001
TFA (25 ml) is added to a solution of tert-butyl 4-(2,4-dioxoimidazolidin-l- yl)piperidine-l-carboxylate (5 g) in 100 ml Of CH2Cl2. After stirring for 5 h at rt, the excess TFA is removed in vacuo to give the title compound (with some inorganic salts) as a white solid which is used without further purification. 1H NMR (300 MHz, DMSOd6) δ 10.80 (IH, s), 8.64 (IH, bs), 8.27 (IH, bs), 3.98 (IH, m), 3.90 (2H, s), 3.30 (2H, m), 2.98 (2H, m), 1.78 (4H, m).
28. fert-Butyl 4-r(2-aminopyridin-3-yl)amino1piperidine-l-carboxylate
Figure imgf000066_0002
Sodium triacetoxyborohydride (29.1 g, 137.4 mmol) is added to a solution of 1,2- diaminopyridine (10 g, 91.6 mmol) and l-Boc-4-piperidone (19.2 g, 96.2 mmol) in 150 ml of DCE at rt. After 5h, another 3.6 g of sodium triacetoxyborohydride is added, followed by an additional 12 h of stirring. The reaction is quenched with 5% aq. NaOH, and the mixture is extracted four times with CH2Cl2. The combined organic extracts are washed with 5% aq. NaOH, water, and brine, dried (Na2SO4), filtered, and evaporated in vacuo to give dark brown oil. Purification by silica gel column chromatography (gradient from CH2Cl2 to 5% MeOH/CH2Cl2/2% Et3N) affords a reddish-brown oil, which is triturated with Et2O to give the title compound as a light brown solid. MS: 293.22 (M+H).
29. fert-Butyl 4-(2-oxo-2,3-dihvdro-lH-imidazor4,5-blpyridin-l-yl)piperidine-l- carboxylate
Figure imgf000066_0003
To a stirred solution of tert-butyl 4-[(2-aminopyridin-3-yl)amino]piperidine-l- carboxylate (10.6 g, 36.3 mmol) in 300 ml of CH3CN at RT is added CDI (6.5 g, 39.9 mmol). After 4 h, an additional 7.5 g of CDI is added, followed by a further 19 h of stirring. The solvent is removed in vacuo, and the brown oily residue is taken up in
CH2Cl2 (200 ml), washed with water (100 ml) and brine (100 ml), dried (Na2SO4), filtered, and evaporated in vacuo to give a gooey brown oil/solid mix. This is dissolved in Et2O, and the product is crystallized by the addition of hexane. Filtration of the solid affords the title compound as a light brown solid. MS: 219.16 (M+H (-Boc)).
30. l-Piperidin-4-yl-l,3-dihvdro-2H-imidazor4,5-blpyridin-2-one bis- hydrochloride
Figure imgf000067_0001
A IM solution of HCl in Et2O (150 ml, 150 mmol) is added at rt to a solution of tert-butyl 4-(2-oxo-2,3-dihydro-lH-imidazo[4,5-b]pyridin-l-yl)piperidine-l-carboxylate (9.6 g, 30.2 mmol) in 230 ml of MeOH. After stirring for 17 h, the reaction mixture is concentrated in vacuo to give the title compound as a light brown solid. MS: 219.16 (M+H).
31. tert-Butyl 4-r(2-nitrobenzyl)amino1piperidine- 1 -carboxylate
Figure imgf000067_0002
To a mixture of 2-nitrobenzylamine hydrochloride (20.0 g, 106.0 mmol) in DCE (250 mL) is added TEA (10.73 g, 106.0 mmol). The mixture is allowed to stir at rt for 5 min. To this mixture N-Boc-4-piperidone (21.13 g, 106.0 mmol), and HOAC (7.96 g, 132.5 mmol), is added and stirred at rt for another 5 min. NaBH(OAc)3 (25.28 g, 119.3 mmol) is added in portions and the mixture is stirred at rt for 1 hr before it is washed with 10 % Na2CO3 solution (100 mL x 3) and brine (50 mL x 2). The organic phase is dried over Na2SO4. The solvent is removed in vacuo to afford the title compound as a red oil, which is used without further purification in subsequent steps.
32. tert-Butyl 4-IY2-aminobenzyl)amino1piperidine-l -carboxvlate
Figure imgf000067_0003
tert-Butyl 4-[(2-nitrobenzyl)amino]piperidine-l-carboxylate (40 g) is dissolved in EtOH (200 mL) and then added 10 % Pd/C (3 g, 2.82 mmol). The mixture is hydrogenated at rt for 2 h. The mixture is passed through celite, the celite is washed with EtOH, and the combined solution is concentrated in vacuo to afford the title compound as a yellow oil, which is used without further purification in subsequent steps.
33. tert-Butyl 4-(2-oxo-l,4-dihvdroquinazolin-3(2H)-yl*)piperidine-l-carboxylate
Figure imgf000068_0001
To a solution of tørt-butyl 4-[(2-aminobenzyl)amino]piperidine-l-carboxylate (30 g) in DCE (250 mL), is added CDI (20.0 g, 122.8 mmol) in portions. The mixture is stirred at rt overnight. The mixture is washed with 1 M citric acid (50 mL x 2), 10 % K2CO3 aqueous solution (50 mL x 3) and dried over MgSO4. The dried extract is filtered and concentrated under vacuum to afford the title compound as a white solid, which is used without further purification in subsequent steps.
34. 3-Piperidin-4-yl-3,4-dihvdroquinazolin-2(lH)-one trifluoroacetate
Figure imgf000068_0002
tert-Butyl 4-(2-oxo-l ,4-dihydroquinazolin-3(2H)-yl)piperidine-l-carboxylate (26.5 g, 80 mmol) is dissolved in CH2Cl2. TFA (61.4 g, 538.5 mmol) is added and the resulting mixture is stirred at rt overnight. The volatiles are removed in vacuo and the resulting solid is rinsed with ether (150 mL χ 2) to afford the title compound as an orange solid. MS: 232.15 (M+).
35. /ert-Butyl 3,3-dimethyl-4-oxopiperidine-l-carboxylate
Figure imgf000068_0003
To a solution of N-Boc-4-piperidone (100 g, 501.9 mmol) in anhydrous THF (2 L) at 0 °C is added NaH (60 % in mineral oil, 42.16 g, 1.05 mol). To this suspension is added iodomethane (178 g, 1.25 mol). The reaction mixture is allowed to stir and warm to rt overnight. The reaction mixture is concentrated in vacuo, ether is added and the resulting mixture is washed with H2O (50 mL x 2) and brine (50 mL x 2), and dried over Na2SO4. The solvent is removed in vacuo and the resulting material is recrystallized from hexanes to afford the title compound as white solid. 36. tgrt-Butyl (5Z)-5-r(dimethylamino)methylenel-3,3-dimethyl-4-oxopiperidine- 1-carboxylate
Figure imgf000069_0001
tert-Buty\ 3,3-dimethyl-4-oxopiperidine-l-carboxylate (5.0 g, 22.0 mmol) is dissolved in dimethylformamide dimethylacetal (22.4 g, 188.2 mmol), and the resulting solution is refluxed for 48 h. The volatiles are removed in vacuo to afford title compound as a green oil, which is used without further purification in subsequent steps. MS: 283.19 (M+).
37. tert-Butyl 7,7-dimethyl-l A6 J-tetrahvdro-5H-pyrazolor4,3-c1pyridine-5- carboxylate
Figure imgf000069_0002
tert-Butyl (5Z)-5-[(dimethylamino)methylene]-3,3-dimethyl-4-oxopiperidine- 1 - carboxylate (6.4 g, 22.7 mmol) is dissolved in anhydrous MeOH (25 mL). Hydrazine (0.80 g, 24.93 mmol) is added and the resulting mixture is stirred at rt overnight. The solvent is removed in vacuo to afford the title compound as a viscous oil.
38. 7J-Dimethyl-4,5,6J-tetrahvdro-lH-pyrazolor4,3-clpyridine dihvdrochloride
Figure imgf000069_0003
2 HCI
To a solution of tert-butyl 7,7-dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3- c]pyridine-5 -carboxylate (40 g, 159 mmol) in DCM (100 mL) is added TFA (80 mL) at 0 °C. The solution is allowed to warm to rt and is stirred at rt for 2 h. The reaction mixture is concentrated in vacuo to give a viscous brown oil. The oil is dissolved in EtOH (90 mL), cooled to 0 °C, concentrated HCl (24 mL) is added and the resulting mixture is concentrated to afford the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 9.74 (2 H, s), 7.61 (1 H, s), 4.05 (2 H, s), 3.14 (2 H, s), 1.35 (6 H, s). MS: 152.14 (M+H+).
39. Ethyl 2-r4-(2-oxo-1.4-dihvdroquinazolin-3(2HVvnpiperidin-l-yl1-1.3- thiazole-4-carboxylate
Figure imgf000070_0001
A mixture of 3-piperidin-4-yl-3,4-dihydroquinazolin-2(lH)-one trifluoroacetate (1.00 g, 2.90 mmol), ethyl 2-chloro-4-thiazolocarboxylate (0.771 g, 4.34 mmol), K2CO3 (1.20 g, 8.70 mmol) and DMA (2 mL) is heated at 150 °C overnight. The mixture is filtered, concentrated in vacuo and purified by silica gel column chromatography to afford the title compound as a white solid, m/z (ES+) 373.12 (M+H+). 40. 2-[4-(2-OxQ- 1 ,4-dihvdroquinazolin-3(2H)-yl)piperidin-l -yli- 1.3-thiazole-4- carboxylic acid
Figure imgf000070_0002
A mixture of ethyl 2-[4-(2-oxo-l,4-dihydroquinazolin-3(2H)-yl)piperidin-l-yl]- l,3-thiazole-4-carboxylate (0.380 g, 1.02 mmol), NaOH (0.20 g, 5.10 mmol), H2O(I mL), and EtOH (6 mL) is reflux ed overnight. The mixture is concentrated in vacuo, HCl (1 N) solution is added to adjust pH to 3. The resulting white solid is collected and washed with water (1 mL x 5) and dried by vacuum to afford the title compound. MS: 359.08 (M+H+).
41. 5-r(4.5-Dichloroisothiazol-3-yl')carbonyll-7J-dimethyl-4.5.6.7-tetrahvdro- lH-pyrazolor4,3-clpyridine
Figure imgf000070_0003
4,5-Dichloroisothiazole-3-carboxylic acid (264 mg, 1.33 mmol), 7,7-dimethyl- 4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine dihydrochloride (250 mg, 1.33 mmol), and TEA (538 mg, 5.32 mmol) are added to DCM (10 mL). The mixture is cooled to 0 0C, and 2-chloro-l,3-dimethylimidazolinium chloride (340 mg, 2.00 mmol) is added and stirred at rt overnight. The mixture is concentrated in vacuo and purified by silica gel column chromatography to afford the title compound. MS: 331.02 (M+). 42. fert-butyl 4-(N-(3 , 5 -dimethoxybenzylV2-bromobenzamido V 5 ,6- dihydropyridine- 1 (2H)-carboxylate
Figure imgf000071_0001
A mixture of N-Boc-piperidone (7.36 g, 36.96 mmol) and 3,5- dimethoxybenzylamine (36.96 mmol) in toluene (60 ml) is refluxed for 4 h with a Dean- Stark trap to remove water formed. Solvent is removed in vacuo, the resulting thick oil is dissolved in toluene (70 ml) and TEA (10.3 ml, 73.92 mmol) is added followed by 2- bromobenzoyl chloride (36.96 mmol). The mixture is heated at 80 0C for 2 h and is then cooled to rt. Water (80 ml) is added, the layers are separated and the aqueous layer is extracted with EtOAc (60 ml). The combined extracts are washed with brine (60 ml), dried over Na2SO4 and evaporated. The residue is purified by silica gel column chromatography to afford the title compound as a light yellow oil. 43. N-Boc-Spiropiperidine intermediate
Figure imgf000071_0002
A mixture of tert-butyl 4-(3,5-dimethoxybenzylamino)-5,6-dihydropyridine-l(2H)- carboxylate (4.63 mmol), Pd(OAc)2 (73 mg, 0.32 mmol, 7 mol%), PPh3 (170 mg, 0.65 mmol, 14 mol%), K2CO3 (959 mg, 6.95 mmol) and Et4NCl (537 mg, 3.24 mmol) in CH3CN (30 ml) is degassed by Ar for 15 min then heated at 110 °C in a sealed tube overnight. Water (40 ml) is added, the layers are separated and the aqueous layer is extracted with EtOAc (40 ml). The combined extracts are washed with brine (40 ml), dried over Na2SO4 and evaporated. The residue is purified by silica gel column chromatography to afford title compound as light yellow oil. 44. Dihydrospiropiperidine intermediate
Figure imgf000072_0001
To solution of the above N-Boc-spiropiperidine intermediate (4 mmol) in dioxane (10 ml) is added HCl solution in dioxane (4N, 15 ml, 60 mmol) and the mixture is stirred at rt overnight. The solvent is removed and to the residue is added aqueous NaHCO3 solution (30 ml) and EtOAc (30 ml). The layers are separated and the aqueous layer is extracted with EtOAc (30 ml). The combined extracts are washed with brine (30 ml), dried over Na2SO4 and evaporated. The residue is purified by silica gel column chromatography to afford the title compound as an off-white solid.
45. 2-(3,5-DimethoxybenzvDspirorisoindole-l,4'-piperidin"l-3(2H)-one
Figure imgf000072_0002
To a solution of the above dihydrospiropiperidine intermediate (3.6 mmol) in
HOAc (20 ml) is added 10% Pd on C (100 mg) and the mixture is hydrogenated at 50 psi overnight. The catalyst is filtered and washed with MeOH (2 >< 10 mL) and the combined filtrate is evaporated to dryness. The residue is partitioned between aqueous NaHCO3 solution (30 ml) and EtOAc (30 ml). The layers are separated and the aqueous layer is extracted with EtOAc (30 ml). The combined extracts are washed with brine (30 ml), dried over Na2SO4 and evaporated. The resulting light yellow solid is used in subsequent reactions without further purification.
46. 2-(3,5-DimethoxybenzylVl'-{4-r(7J-dimethyl-1.4.6.7-tetrahvdro-5H- pyrazolor4,3-c1pyridin-5-yl)carbonyllpyridin-2-yl)spirorisoindole-l,4'-piperidin1-3(2HV one
Figure imgf000073_0001
5-(2-Chloroisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3- cjpyridine (100 mg, 0.344 mmol), 2-(3,5-dimethoxybenzyl)spiro[isoindole-l,4'-piperidin]- 3(2H)-one (121.2 mg, 0.344 mmol), and K2CO3 (190 mg, 1.38 mmol) are added to DMA (1 mL) and the resulting mixture is heated at 150 °C overnight. The mixture is diluted with DCM (30 mL), washed with water (5 mL x 3) and brine (5 mL x 2), and dried over MgSO4. The solvent is removed in vacuo and purified by silica gel column chromatography to afford title compound. MS: 607.50 (M+).
47. 6'-f 3.5-Dimethoxybenzvn-l- {4-IY7.7-dimethyl-l .4.6.7-tetrahvdro-5H- pyrazolor4.3-clpyridin-5-vDcarbonyllpyridin-2-yl}spirorpiperidine-4,7'-pyrrolor3.4- blpyridinl-5Y6'HVone
Figure imgf000073_0002
5-(2-Chloroisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3- c]pyridine (100 mg, 0.344 mmol), 6'-(3,5-dimethoxybenzyl)spiro[piperidine-4,7'- pyrrolo[3,4-b]pyridin]-5'(6Η)-one (122 mg, 0.344 mmol) and K2CO3 (190 mg, 1.376 mmol) are added to DMA (1 mL), and the resulting mixture is heated at 150 °C overnight. The mixture is diluted with DCM (30 mL), washed with water (5 mL * 3) and brine (5 mL x 2), and dried over MgSO4. The solvent is removed in vacuo and purified by silica gel column chromatography to give the title compound as yellow solid. MS: 608.48 (M+). 48. 2-Chloro-4-r(4-pyridin-4-ylpiperidin-l-vDcarbonyl1pyridine
Figure imgf000073_0003
4-Piperidin-4-ylpyridine (324 mg, 2.0 mmol) is added to DCE (1 mL), and the resulting mixture is cooled to 0 °C. To this suspension is added a mixture of 2- chloroisonicotinoyl chloride (352 mg, 2.0 mmol) and CH2Cl2 (10 mL). The resulting mixture is stirred at 0 0C to rt overnight. H2O (10 mL) is added and the resulting mixture is extracted with CH2Cl2 (30 mL * 3). The extracts are dried over MgSO4 and solvent is removed in vacuo. Silica gel column chromatography affords the title compound as yellow solid. MS: 302.09 (M+).
49. l-Benzyl-l'-(4-r(7J-dimethyl-l,4.6.7-tetrahvdro-5H-pyrazolor4,3-clpyridin -5 - vDcarbonyllp yridin-2-yl| spiro [indole- 3 ,4'-piperidinl-2( 1 HVone
Figure imgf000074_0001
5-(2-Chloroisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3- c]pyridine (123 mg, 0.423 mmol), l-benzylspiro[indole-3,4'-piperidin]-2(lH)-one hydrochloride (153 mg, 0.465 mmol), and K2CO3 (183 mg, 1.324 mmol) are added to
DMA (1.0 mL), and the resulting mixture is heated at 150 0C overnight. DCM (30 mL) is added, and the mixture is washed with water (5 ml x 3) and brine (5 mL x 2), and dried over MgSO4. The solvent is removed in vacuo and silica gel column chromatography affords the title compound as light yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.49 (1 H, m), 8.27 (1 H, m), 7.24-7.39 (8 H, m), 7.17 (1 H, m), 7.02 (1 H, m), 6.74 (2 H, m), 6.58 (1 H, m), 4.92 (1 H, s), 4.83 (1 H, m), 4.44 (1 H, m), 4.00-4.14 (2 H, m), 3.79 (1 H, s), 3.49 (1 H, m), 3.33 (1 H, s), 1.91-2.05 (4 H, m), 1.40 (3 H, s), 1.27 (3 H, s). MS: 547.42 (M+).
50. 5-(2-Chloro-6-methoxyisonicotinoylV7J-dimethyl-4.5,6,7-tetrahydro-lH- pyrazolor4,3-clpyridine
Figure imgf000074_0002
7,7-Dimethyl-4,5,6,7-tetrahydro- lH-pyrazolo[4,3-c]pyridine dihydrochloride (2.18 g, 9.71 mmol) and TEA (4.91 g, 48.6 mmol) are added to DCM (10 mL), and the resulting mixture is cooled to 0 °C. To this suspension is added a mixture of 2-chloro-6- methoxyisonicotinoyl chloride (2.00 g, 9.71 mmol) and DCM (10 mL) dropwise. The resulting mixture is stirred at 0 °C to rt overnight. The mixture is diluted with H2O (10 mL) and the resulting mixture is extracted with DCM (15 mL x 3). The extracts are dried over MgSO4, solvent is removed in vacuo and silica gel column chromatography affords the title compound as light yellow solid. MS: 321.10 (M+).
51. tert-Butyl 4-oxo-9-azadispiror2.1.2.31decane-9-carboxylate
Figure imgf000075_0001
t-BuOK (24.7 g, 219.8 mmol) is dissolved in t-BuOH (350 mL) after heating at 60
°C. To this solution, N-Boc-4-piperidone (43.8 g, 219.8 mmol) is added and the resulting mixture is stirred at rt for 1 h. 2-Chloroethyl dimethylsulfonium iodide (50.0 g, 198.0 mmol) is added in portions and the resulting mixture is stirred at rt for 2 h. To this mixture is added a solution of t-BuOK (24.7 g, 219.8 mmol) in t-BuOH (350 mL) and the resulting mixture is allowed to stir at rt for 48 h. The reaction mixture is poured into H2O (2.5 L), and the resulting mixture is extracted with EtOAc (500 mL x 5) and dried over MgSO4. The solvent is removed in vacuo, and silica gel column chromatography affords the title compound as a white solid.
52. 9-Azadispiror2.1.2.31decan-4-one
Figure imgf000075_0002
tert-BvXy\ 4-oxo-9-azadispiro[2.1.2.3]decane-9-carboxylate (1.0 g, 3.98 mmol) is dissolved in DCM (5 mL) and cooled to 0 °C. TFA (4.61 g, 40.4 mmol) is added and the resulting solution is stirred at 0 °C to rt for 2 h. Volatiles are removed in vacuo, and DCM (50 mL) is added. To this solution is added saturated NaHCO3 aqueous solution (10 mL) and the mixture is stirred at 0 °C for 30 min. The organic phase is separated and the aqueous phase is extracted with DCM (30 mL x 4). The organic phase is combined, dried over MgSO4, and solvent is removed in vacuo. The title compound is obtained as dark yellow solid. 1H NMR (400 MHz, CDCl3) δ (1 H, s), 3.44 (4 H, s), 1.48 (m, 4 H), 0.98 (m, 4 H). 53. 9-(2-Chloroisonicotinoyl)-9-azadispiro[2.1 ,2.31decan-4-one
Figure imgf000075_0003
2-Chloroisonicotonic acid (0.208 g, 1.32 mmol), 9-azadispiro[2.1.2.3]decan-4-one (0.200 g, 1.32 mmol), and TEA (534 mg, 5.28 mmol) are added to DCM (4 mL). The mixture is cooled to 0 0C, and 2-chloro-l,3-dimethylimidazolinium chloride (335 mg, 1.98 mmol) is added. The reaction is stirred at 0 °C to rt overnight. The solvent is removed in vacuo. Purification by silica gel column chromatography affords the title compound. MS: 291.12 (M+).
54. tert-Butyl 8-oxo-5-azaspiror2.51octane-5-carboxylate
Figure imgf000076_0001
t-BuOK (24.7 g, 219.8 mmol) is dissolved in t-BuOH (350 mL) after heating at 60 0C. To this solution, N-Boc-4-piperidone (43.8 g, 219.8 mmol) is added and the resulting mixture is stirred at rt for 1 h. 2-Chloroethyldimethylsulfonium iodide (50.0 g, 198.0 mmol) is added in portions and the resulting mixture is stirred at rt for 2 h. To this mixture is added a solution of t-BuOK (24.7 g, 219.8 mmol) in t-BuOH (350 mL) and the resulting mixture is allowed to stir at rt for 48 h. The reaction mixture is poured into H2O (2.5 L), and the resulting mixture is extracted with EtOAc (500 mL x 5) and dried over MgSO4. The solvent is removed in vacuo, and silica gel column chromatography affords the title compound as a waxy solid. 1H NMR (400 MHz, CDCl3) δ 3.78 (2 H, m), 3.57 (2 H, broad s), 2.60 (2 H, t), 1.48 (9 H, s), 1.30-1.26 (4 H, m).
55. tert-Butyl (7Z)-7-r(dimethylamino')methylenel-8-oxo-5-azaspiror2.51octane- 5-carboxvlate
Figure imgf000076_0002
o tert-Butyl 8-oxo-5-azaspiro[2.5]octane-5-carboxylate is dissolved in dimethyl formamide dimethylacetal (17.98 g, 150.9 mmol), and the resulting solution is refluxed for 26 h. Volatiles are removed in vacuo and the title compound is obtained as a green oil, which is used without further purification in subsequent steps.
56. tert-Butyl l\4'-dihvdrospirorcvclopropane-l,7'-pyrazolor4,3-c1pyridine'|- 5'(6'HVcarboxylate
Figure imgf000076_0003
tert-Butyl (7Z)-7-[(dimethylamino)methylene]-8-oxo-5-azaspiro[2.5]octane-5- carboxylate (4.55 g, 15.09 mmol) is dissolved in anhydrous MeOH (17 mL). Hydrazine (0.532 g, 16.6 mmol) is added and the resulting mixture is stirred at rt overnight. Solvents are removed in vacuo and silica gel column chromatography affords the title compound as a red oil. MS: 250.19 (M+H+).
57. 1\4\5\6'-Tetrahvdrospiro[cvclopropane-1 J'-pyrazoloKJ-cipyridinei dihvdrochloride
Figure imgf000077_0001
tert-Butyl r,4'-dihydrospiro[cyclopropane-l,7'-pyrazolo[4,3-c]pyridine]-5'(6Η)- carboxylate (1.2 g, 4.81 mmol) is dissolved in EtOAc (35 mL). The solution is cooled to 0 0C and HCl (4 N) in dioxane is added dropwise. The mixture is stirred at 0 °C to rt overnight. The precipitate is filtered, washed with EtOAc (2 mL x T), and dried. The title compound is obtained as white solid. NMR (400 MHz, CD3OD) δ 7.88 (1 H, m), 4.42 (2 H, s), 3.44 (2 H, s), 1.36 (m, 2 H), 1.26 (2 H, m). MS: 150.23 (M+H).
58. 5'-(2-Chloroisonicotinoyl)-r,4',5',6'-tetrahvdrospirorcyclopropane-l,7'- p yrazolo [4,3 -cipyridinei
Figure imgf000077_0002
r,4',5',6'-Tetrahydrospiro [cyclopropane- l,7'-pyrazolo[4,3-c]pyridine] dihydrochloride (222 mg, 1.0 mmol) and TEA (506 mg, 5.0 mmol) are added to DCM (2 mL), and the resulting mixture is cooled to 0 °C. To this suspension is added dropwise a mixture of 2-chloroisonicotinoyl chloride (176 mg, 1.0 mmol) dissolved in DCM (2 mL). The resulting mixture is stirred at 0 0C to rt overnight and then diluted with H2O (10 mL), and the resulting mixture is extracted with DCM (15 mL x 3). The extracts are dried over MgSO4 and solvent is removed in vacuo. Silica gel column chromatography affords the title compound as a light yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.48 (1 H, d), 2.39 (1 H, d), 7.26 (2 H, m), 4.91 (1 H, s), 4.51 (1 H, s), 3.88 (1 H, s), 3.42 (1 H, s), 1.21 (2 H, m), 1.11 (1 H, m), 0.70 (1 H, m). MS: 289.08 (M+).
59. l-Benzyl-4-(2-chloroquinolin-3-yl)piperidin-4-ol
Figure imgf000077_0003
A solution of n-BuLi in hexane (1.6 M, 76.4 ml, 122 mmol) is added dropwise to a solution of diisopropylamine (17.2 ml, 122 mmol) in 250 ml of THF at -78 °C under N2. After stirring for 1 h, a solution of 2-chloroquinoline (20 g, 122 mmol) in 50 ml of THF is added via a dropping funnel over 30 min. After a further 1 h, a solution of 1- benzyl-4-piperidone (22.6 ml, 122 mmol) in 10 ml of THF is added to the brown reaction mixture, and stirring is continued for an additional 40 min. The reaction is then allowed to slowly warm to rt. After cooling the mixture back to -20 °C, the reaction is carefully quenched with water (100 ml). The resulting mixture is extracted with EtOAc, and the organic phase is washed with brine, dried (Na2SO4), filtered and evaporated. Silica gel chromatographic purification (gradient from CH2Cl2 to 10% [5%
NH4OH/MeOH]/CH2Cl2) affords the title compound. MS: 353.15 (M+H). 60. 3-(l-Benzyl-L2,3,6-tetrahvdropyridin-4-yl)quinolin-2(lH)-one
Figure imgf000078_0001
A mixture of l-benzyl-4-(2-chloroquinolin-3-yl)piperidin-4-ol (22 g, 62.3 mmol) in 300 ml of 6N HCl is stirred under reflux for 1O h. The reaction mixture is cooled to rt and diluted with water (200 ml). The precipitated solid is collected and dried in vacuo to give the title compound. Mass spec. 317.18 (M+H). 61. 3 -Piperidin-4- ylquinolin-2( 1 HVone
Figure imgf000078_0002
A mixture of 3-(l-benzyl-l,2,3,6-tetrahydropyridin-4-yl)quinolin-2(lH)-one (4.0 g, 12.6 mmol) in 200 ml of MeOH is degassed with N2 (g) for 5 minutes, and 10% Pd/C (1.2 g) is added. The resulting reaction mixture is hydrogenated under 30 psi of H2 (g) at 50 °C for 18 h on a Parr shaker. The mixture is filtered through celite, and the celite is washed well with MeOH. The filtrate is concentrated in vacuo to give the title compound as a white solid. MS: 229.16 (M+H).
62. N'-Phenyl-2-pyridin-4-ylacetohvdrazide
Figure imgf000079_0001
To a solution of pyridin-4-ylacetic acid hydrochloride (20 g, 115 mmol) in 130 ml of DMF at rt is added l-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (26.5 g, 138 mmol), 1-hydroxybenzotriazole (18.6 g, 138 mmol), diisopropylethylamine (60 ml, 345 mmol), and phenylhydrazine (11.3 ml, 115 mmol). After stirring for 22 h, the reaction mixture is diluted with CH2Cl2 (300 ml) and washed with sat. NaHCO3 (300 ml). The aqueous phase is extracted twice more with CH2Cl2 (100 ml each), and the combined organic extracts are dried (Na2SO4), filtered and evaporated in vacuo to give a red solid. This solid is suspended in EtOAc and heated to 50 0C for 15 min with vigorous stirring. After cooling, the solid is collected by vacuum filtration and dried in vacuo to give the title compound as an off-white solid. MS: 228.18 (M+H).
Figure imgf000079_0002
Calcium hydride (2.8 g, 67.5 mmol) is added to a DMF solution (100 ml) of N'- phenyl-2-pyridin-4-ylacetohydrazide (9.3 g, 40.9 mmol), and the reaction mixture is heated to 170 °C and stirred for 14 h. After cooling to rt, the mixture is filtered, and the filtrate is concentrated in vacuo to give a reddish-brown residue. This residue is dissolved in CH2Cl2, and hexane is added with stirring until a solid precipitates from the solution. The solid is collected by filtration and dried in vacuo to give the title compound as a brown solid. MS: 238.17 (M+H).
64. 1 -Phenyl-4-piperidin-4-yl- 1 ,2-dihvdro-3H-pyrazol-3-one
Figure imgf000079_0003
Platinum oxide (3.4 g) is added to a mixture of l-phenyl-4-pyridin-4-yl-l,2- dihydro-3H-pyrazol-3-one (9.7 g, 40.9 mmol) in 120 ml of glacial AcOH, 1.0 ml of cone. HCl, and 50 ml of water. The resulting mixture is purged with N2 (g) for 5-10 min, and then hydrogenated under 40-50 psi of H2 (g) on a Parr shaker for 36 h at rt. The mixture is filtered through celite, and the filtrate is concentrated in vacuo to give a gooey, yellow solid. Trituration with Et2O affords the title compound as a yellow solid. MS: 244.12 (M+H).
65. 9H-Fluoren-9-ylmethyl 4-r(tert-butoxycarbonyl')hvdrazono1piperidine- 1 - carboxylate
Figure imgf000080_0001
To a solution of l-Fmoc-4-piperidone (25 g, 78 mmol) in 390 ml of EtOH is added tert-butyl carbazate (11.3 g, 86.6 mmol), and the reaction mixture is heated to reflux and stirred forl h. The solution is cooled to rt and concentrated in vacuo to give white foam. Et2O (150 ml) is added, and the solution is stirred at rt for 30 min, at which time a solid precipitates from the solution. The solid is collected by vacuum filtration and dried in vacuo to give the title compound as a white solid
66. 9H-Fluoren-9-ylmethyl 4-r2-(tert-butoxycarbonyl)hvdrazinolpiperidine-l- carboxylate
Figure imgf000080_0002
Platinum oxide (3.3 g) is added to a solution of 9H-fluoren-9-ylmethyl Λ-[(tert- butoxycarbonyl)-hydrazono]piperidine-l-carboxylate (32.7 g, 75 mmol) in acetic acid (200 ml). The resulting mixture is hydrogenated on a Parr shaker under 45 psi of H2(g) for 2 h. The solution is filtered through celite, and the filtrate is concentrated in vacuo to give the title compound.
67. 9H-Fluoren-9- ylmethyl 4-hvdrazinopiperidine- 1 -carboxylate trifluoroacetate
Figure imgf000080_0003
A solution of 9H-fluoren-9-ylmethyl 4-[2-{tert- butoxycarbonyl)hydrazino]piperidine-l-carboxylate (32 g, 73 mmol) in 160 ml of trifluoroacetic acid is stirred at rt for 3 h. The mixture is concentrated in vacuo to give the title compound as reddish-brown viscous oil, which is suitable for further use without any additional purification.
68. Ethyl (phenylcarbonothioylicarbamate
Figure imgf000081_0001
Aluminum chloride (13.3 g, 100 mmol) is added portionwise to an ice-cold solution of ethoxycarbonyl isothiocyanate (5.9 ml, 50 mmol) and benzene (4.4 ml, 50 mmol) in 30 ml OfCH2Cl2. The reaction mixture is stirred at 0 0C for 4 h, and then it is quenched by careful addition of cold IN HCl. Enough CH2Cl2 is added to dissolve any solid organic material, and the resulting mixture is extracted with 10% NaOH (4 x 50 ml). The combined aqueous phases are further basified to pH=l 1-12 to dissolve all the solids in the mixture. This basic aqueous solution is extracted with Et2O and acidified to pH=3-4 with cone. HCl while cooling in an ice bath. The cloudy mixture is extracted with CH2Cl2 (4 x), and the combined organic extracts are dried (Na2SO4), filtered and evaporated in vacuo to give a red oil. Purification by silica gel column chromatography (gradient from Hex to 30% EtO Ac/Hex) affords red oil, which is dissolved in Et2O and cooled in a dry- ice/acetone bath. Hexane is slowly added while scratching the flask with a spatula, and eventually a solid precipitates from solution. This solid is collected by vacuum filtration and dried in vacuo to give the title compound as a yellow solid. MS: 210.08 (M+H). 69. Ethyl r(3-methoxyphenyl)carbonothioyllcarbamate
Figure imgf000081_0002
A solution of ethoxycarbonyl isothiocyanate (5.0 g, 38.1 mmol) in 20 ml of THF is cooled to 0 °C and treated with 3-methoxyphenylmagnesium bromide (IM in THF, 38.1 ml, 38.1 mmol) by dropwise addition. The resulting mixture is stirred for 2 h at 0 °C, quenched with 15-20 ml of IN HCl, and diluted with CH2Cl2 and 20% NaOH (100 ml each). The resulting mixture is filtered through celite, and the filtrate is transferred to a separatory funnel. The layers are separated, and the organic layer is extracted twice more with 20% NaOH (50 ml each). The combined aqueous extracts are washed with Et2O (2 x 75 ml) and acidified to pH=3-4 with cone. HCl while cooling in an ice bath. The cloudy mixture is extracted two times with CH2Cl2, and the combined organic extracts are dried (Na2SO4), filtered and evaporated in vacuo to give a red oil. Purification by silica gel column chromatography (gradient from Hex to 30% EtOAc/Hex) affords the title compound as red oil. MS: 240.10 (M+H).
70. 9H-Fluoren-9-ylmethyl 4-('5-oxo-3-phenyl-4.5-dihvdro-lH-1.2.4-triazol-l- yl)piperidine- 1 -carboxylate
Figure imgf000082_0001
A solution of 9H-fluoren-9-ylmethyl 4-hydrazinopiperidine-l-carboxylate trifluoroacetate (3.0 g, 6.65 mmol), ethyl (phenylcarbonothioytycarbamate (1.5 g, 7.32 mmol), and diisopropylethylamine (1.3 ml, 7.32 mmol) in 30 ml of THF is stirred under reflux for 48 h (at the end of the first day, an additional 3.0 g of 9H-fluoren-9-ylmethyl 4- hydrazinopiperidine-1-carboxylate trifluoroacetate is added). After cooling to rt, the reaction is concentrated to give a reddish-brown oil. This oil is dissolved in acetonitrile with heating, and then cooled back to rt. The resulting precipitate is filtered and dried in vacuo to give the title compound as an off- white solid. MS: 467.14 (M+H).
71. 9H-Fluoren-9-ylmethyl 4-r3-(3-methoxyphenvn-5-oxo-4,5-dihvdro-lH-1.2.4- triazol- 1 - yllpiperidine- 1 -carboxylate
Figure imgf000082_0002
A solution of 9H-fluoren-9-ylmethyl 4-hydrazinopiperidine-l-carboxylate trifluoroacetate (3.0 g, 6.65 mmol), ethyl [(3-methoxyphenyl)carbonothioyl]carbamate (1.75 g, 7.32 mmol), and diisopropylethylamine (1.3 ml, 7.32 mmol) in 30 ml of THF is stirred under reflux for 72 h (at the end of the first day, an additional 3.0 g of 9H-fluoren- 9-ylmethyl 4-hydrazinopiperidine-l-carboxylate trifluoroacetate is added). After cooling to rt, the reaction is concentrated to give a brown oil/solid mix. This crude material is dissolved in acetonitrile with heating and filtered. The collected solid is dried in vacuo to give the title compound as a white solid. MS: 497.16, (M+H).
72. 5-Phenyl-2-piperidin-4-yl-2,4-dihvdro-3H-1.2.4-triazol-3-one
Figure imgf000083_0001
A solution of 9H-fluoren-9-ylmethyl 4-(5-oxo-3-phenyl-4,5-dihydro-lH-l ,2,4- triazol-l-yl)piperidine-l-carboxylate (1.25 g, 2.68 mmol) and diethylamine (10 ml) in THF (10 ml) is stirred for 21 h at rt. The mixture is concentrated, and the residue is purified by silica gel column chromatography (gradient from CH2Cl2 to 10%[5% NH40H/Me0H]/ CH2Cl2) to give the title compound as a white solid. MS: 245.21 (M+H).
73. 5-(3-Methoxyphenyl)-2-piperidm-4-yl-2,4-dihvdro-3H-l,2.4-triazol-3-one
Figure imgf000083_0002
A solution of 9H-fluoren-9-ylmethyl 4-[3-(3-methoxyphenyl)-5-oxo-4,5-dihydro- lH-l,2,4-triazol-l-yl]piperidine-l-carboxylate (1.39 g, 2.80 mmol) and diethylamine (15 ml) in THF (15 ml) is stirred for 1 week at rt. The mixture is concentrated, and the residue is purified by silica gel column chromatography (gradient from CH2Cl2 to 15%[5% NH40H/Me0H]/ CH2Cl2) to give the title compound as a white solid. MS: 275.16 (M+H).
EXAMPLE 2. SYNTHESIS OF REPRESENTATIVE AMIDE-SUBSTITUTED ARYL PIPERIDINES This Example illustrates the synthesis of certain representative amide-substituted aryl piperidines. All amide-substituted aryl piperidines in this Example are CGRP receptor antagonists, and all except Example 2E have an IC50 (determined as described in Example 5) that is 2 micromolar or less (i.e., the concentration of such compounds that is required to provide a 50% decrease in the fluorescence response of cells exposed to one IC50 of CGRP is 2 micromolar or less). A. N,4-Dimethyl-N-[2-methyl-2-(lH-pyrazol-5-yl)propyl]-2-[4-(2-oxo-l,2,4,5- tetrahydro-3H-l,3-benzodiazepm-3-yl)piperidin-l-yl]-l,3-thiazole-5-carboxamide
Figure imgf000084_0001
A mixture of 3-piperidin-4-yl-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one trifluoroacetate (356 mg, 0.99 mmol), tert-buty\ 5-(2-{[(2-chloro-4-methyl-l,3-thiazol-5- yl)carbonyl] (methyl)-amino } - 1 , 1 -dimethylethyl)- 1 H-pyrazole- 1 -carboxylate (410 mg, 0.99 mmol), and potassium carbonate (410 mg, 2.97 mmol) in 4 mL of dioxane is heated to 100 0C in a sealed tube. After 48 h, the reaction is cooled to rt and the dioxane is removed in vacuo. The residue is purified by silica gel column chromatography (gradient from 50% EtOAc/CH2Cl2 to 5% MeOH/EtOAc) to give the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.51 (IH, s), 7.10 (2H, m), 6.90 (IH, t, J 7.2), 6.72 (IH, d, J7.6), 6.61 (IH, bs), 6.19 (IH, s), 4.49 (IH, m), 4.06 (2H, d, J 13.6), 3.77 (2H, s), 3.44 (2H, m), 3.09 (2H, m), 2.97 (2H, m), 2.74 (3H, s), 2.21 (3H, s), 1.78 (4H, m), 1.38 (6H, s). MS: 522.27 (M+H). B. 3-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-methylpyridin-2-l}piperidin-4-yl)-l,3,4,5-tetrahydro-2H-l,3- benzodiazepin-2-one
Figure imgf000084_0002
A mixture of 5-(2-chloro-6-methylisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro- lH-pyrazolo[4,3-c]pyridine (200 mg, 0.66 mmol), 3-piperidin-4-yl-l,3,4,5-tetrahydro-2H- l,3-benzodiazepin-2-one trifluoroacetate (237 mg, 0.66 mmol), and potassium carbonate (274 mg, 1.98 mmol) in 1 ml of NMP is heated to 150 0C and stirred for 24 h in a sealed tube. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50% EtOAc/CH2Cl2 to 2% MeOH/EtOAc) to give the title compound as an off-white solid. 1H NMR (400 MHz, CDCl3) δ 7.38 (IH, s), 7.22 (IH, s), 7.10 (2H, m), 6.90 (IH, t, J7.6), 6.72 (IH, d, J8), 6.60 (IH, s), 6.43 (IH, m), 4.80 (IH, s), 4.50 (4H, m), 3.77 (IH, s), 3.43 (2H, m), 3.37 (IH, s), 2.95 (4H, m), 2.41 (3H, d, J 6.4), 1.8 (4H, m), 1.40 (3H, s), 1.24 (3H, s). MS: 514.35 (M+H). C. l-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-ρyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-methylpyridin-2-l}piperidin-4-yl)-4-phenyl-l,3-dihydro-2H-imidazol-2- one
Figure imgf000085_0001
A mixture of 5-(2-chloro-6-methylisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro- lH-pyrazolo[4,3-c]pyridine (100 mg, 0.33 mmol), 4-phenyl-l-piperidin-4-yl-l,3-dihydro- 2H-imidazol-2-one (80 mg, 0.33 mmol), and potassium carbonate (137 mg, 0.99 mmol) in 1 mL of NMP is heated to 150 0C and stirred for 14 h in a sealed tube. An additional 80 mg of 4-phenyl-l-piperidin-4-yl-l,3-dihydro-2H-imidazol-2-one is added, followed by another 9 h at 150 °C. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50% EtOAc/CH2Cl2 to 6% MeOH/EtOAc) to give the title compound as a pale brown solid. 1H NMR (400 MHz, DMSOd6) δ 10.69 (IH, s), 7.48 (2H, d, J7.2), 7.29 (3H, t, J7.6), 7.15 (2H, m), 6.61 (IH, s), 6.41 (IH, s), 4.60 (IH, bs), 4.50 (2H, m), 4.34 (IH, bs), 4.12 (IH, m), 3.62 (IH, s), 2.91 (2H, m), 2.32 (3H, d, J4.4), 1.78 (5H, m), 1.24 (3H, s), 1.12 (3H, s). MS: 512.31 (M+H).
D. l-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-methylpyridin-2-l}piperidin-4-yl)-l,3-dihydro-2H-benzimidazol-2-one
Figure imgf000085_0002
A mixture of 5-(2-chloro-6-methylisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro- lH-pyrazolo[4,3-c]pyridine (100 mg, 0.33 mmol), l-piperidin-4-yl-l,3-dihydro-2H- benzimidazol-2-one (72 mg, 0.33 mmol), and potassium carbonate (137 mg, 0.99 mmol) in 1 ml of NMP is heated to 150 °C and stirred for 14 h in a sealed tube. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50% EtOAc/CH2Cl2 to 3% MeOH/EtOAc) to give the title compound as an off-white solid. 1H NMR (400 MHz, CDCl3) δ 9.43 (IH, bs), 7.39 (IH, s), 7.24 (IH, s), 7.07 (4H, m), 6.49 (2H, m), 4.81 (IH, s), 4.61 (3H, m), 4.45 (IH, s), 3.78 (IH, s), 3.40 (IH, s), 2.98 (2H, m), 2.44 (5H, m), 1.93 (2H, m), 1.41 (3H, s), 1.27 (3H, s). MS: 486.28 (M+H).
E. 8-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]pyridin-2-yl}-l-phenyl-l,3,8-triazaspiro[4.5]decan-4-one
Figure imgf000086_0001
A mixture of 5-(2-chloroisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro-lH- pyrazolo[4,3-c]pyridine (130 mg, 0.45 mmol), l-phenyl-l,3,8-triazaspiro[4.5]decan-4-one (104 mg, 0.45 mmol), and potassium carbonate (124 mg, 0.90 mmol) in 1 ml of NMP is heated to 150 °C and stirred for 15 h in a sealed tube. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50%
EtOAc/CH2Cl2 to 1% MeOH/EtOAc) to give the title compound as a white solid. 1H NMR (300 MHz, CDCl3) δ 8.27 (IH, d, J 5.1), 7.38 (IH, s), 7.16 (2H, m), 6.81 (2H, m), 6.65 (4H, m), 4.81 (IH, s), 4.76 (2H, s), 4.43 (IH, s), 4.26 (2H, m), 3.78 (3H, m), 3.39 (IH, s), 2.65 (2H, m), 1.80 (2H, m), 1.39 (3H, s), 1.23 (3H, s). MS: 486.32 (M+H). F. 3-(l-{2-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]pyridin-4-yl}piperidin-4-yl)-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one
Figure imgf000086_0002
A mixture of 5-[(4-chloropyridin-2-yl)carbonyl]-7,7-dimethyl-4,5,6,7-tetrahydro- lH-pyrazolo[4,3-c]pyridine (90 mg, 0.31 mmol), 3-piperidin-4-yl-l,3,4,5-tetrahydro-2H- l,3-benzodiazepin-2-one trifluoroacetate (111 mg, 0.31 mmol), and potassium carbonate (129 mg, 0.93 mmol) in 1 mL of NMP is heated to 150 °C and stirred for 3 days in a sealed tube. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from CH2Cl2 to 6% MeOH/ CH2Cl2) to give the title compound as a light brown solid. 1H NMR (300 MHz, CDCl3) δ 8.26 (IH, d, J6), 7.37 (IH, s), 7.22 (IH, s), 7.04 (2H, m), 6.90 (IH, t, J 7.2), 6.74 (IH, m), 6.55 (IH, s), 4.86 (IH, s), 4.66 (IH, s), 4.53 (IH, m), 4.02 (2H, d, J 13.5), 3.80 (IH, s), 3.64 (IH, s), 3.42 (2H, m), 3.0 (4H, m), 1.78 (4H, m), 1.40 (3H, s), 1.22 (3H, s). MS: 500.41 (M+H). G. 3-(l-{5-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-4-methyl-l,3-thiazol-2-yl}piperidin-4-yl)-l,3,4,5-tetrahydro-2H-l,3- benzodiazepin-2-one
Figure imgf000087_0001
A mixture of 5-[(2-chloro-4-methyl-l,3-thiazol-5-yl)carbonyl]-7,7-dimethyl- 4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridine (100 mg, 0.32 mmol), 3-piperidin-4-yl- l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one trifluoroacetate (115 mg, 0.32 mmol), and potassium carbonate (133 mg, 0.96 mmol) in 1 ml of NMP is heated to 150 °C and stirred for 2 h in a sealed tube. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50%EtOAc/CH2Cl2 to 6% MeOH/ EtOAc) to give the title compound as an off-white solid. 1H NMR (400 MHz, CDCl3) δ 7.32 (IH, s), 7.09 (2H, m), 6.91 (IH, m), 6.73 (IH, d, J 8), 6.61 (IH, bs), 4.71 (2H, s), 4.51 (IH, m), 4.09 (2H, m), 3.66 (2H, s), 3.47 (2H, m), 3.14 (2H, m), 2.98 (2H, m), 2.25 (3H, s), 1.80 (4H, m), 1.32 (6H, s). MS: 520.38 (M+H).
H. 3-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-(methoxymethyl)pyridin-2-yl}piperidin-4-yl)-l,3,4,5-tetrahydro-2H-l,3- benzodiazepin-2-one
Figure imgf000087_0002
A mixture of 5-[2-chloro-6-(methoxymethyl)isonicotinoyl]-7,7-dimethyl-4,5,6,7- tetrahydro-lH-pyrazolo[4,3-c]pyridine (2.0 g, 5.97 mmol), 3-piperidin-4-yl-l,3,4,5- tetrahydro-2H-l,3-benzodiazepin-2-one trifluoroacetate (2.1 g, 5.97 mmol), and diisopropylethylamine (3.1 ml, 17.91 mmol) in 10 mL of NMP is heated to 150 °C and stirred for 20 h in a sealed tube. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50%EtOAc/CH2Cl2 to 6% MeOH/ EtOAc) to give the title compound as a light brown solid. 1H NMR (400 MHz, CDCl3) δ 7.38 (IH, s), 7.21 (IH, s), 7.10 (IH, t), 7.04 (IH, d, J7.6), 6.89 (IH, t, J7.6), 6.71 (2H, m), 6.55 (2H, m), 4.81 (IH, s), 4.49 (6H, m), 3.78 (IH, s), 3.44 (6H, m), 2.97 (4H, m), 1.82 (2H, m), 1.71 (2H, m), 1.40 (3H, s), 1.24 (3H, s). MS: 544.35 (M+H).
I. 3-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-(hydroxymethyl)pyridin-2-yl}piperidin-4-yl)-l,3,4,5-tetrahydro-2H-l,3- benzodiazepin-2-one
Figure imgf000088_0001
A solution of 3-(l-{4-[(7,7-dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3- c]pyridin-5-yl)carbonyl]-6-(methoxymethyl)pyridin-2-yl}piperidin-4-yl)-l, 3,4,5- tetrahydro-2H-l,3-benzodiazepin-2-one (200 mg, 0.37 mmol) in 10 ml OfCH2Cl2 is cooled to -78 °C and treated with boron tribromide (IM in CH2Cl2, 1.85 ml, 1.85 mmol) by dropwise addition. After 30 min at -78 0C, the reaction is warmed to rt and stirred for an additional 1 h. Methanol (2.0 ml) is added, and the mixture is concentrated in vacuo. The residue is partitioned between EtOAc and sat. NaHCO3, and the layers are separated. The organic layer is dried (Na2SO4), filtered and evaporated, and the residue is purified by silica gel column chromatography (gradient from CH2Cl2 to 5% MeOH/CH2Cl2 /2% Et3N) to afford the title compound as an off-white solid. 1H NMR (400 MHz, CDCl3) δ 7.39 (IH, s), 7.23 (IH, s), 7.11 (IH, t, J7.6), 7.05 (IH, d, J6.8), 6.90 (IH, t, J7.2), 6.72 (IH, d, J 8), 6.50 (3H, m), 4.81 (IH, s), 4.64 (2H, d, J7.2), 4.48 (4H, m), 3.78 (IH, s), 3.44 (2H, m), 3.37 (IH, s), 2.97 (4H, m), 1.85 (2H, m), 1.70 (2H, m), 1.40 (3H, s), 1.25 (3H, s). MS: 530.25 (M+H).
J. 3-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl] -6-(4-pyridin-4-ylpiperidin- 1 -yl)p yridin-2-yl} piperidin-4-yl)- 1 ,3 ,4,5- tetrahydro-2H-l,3-benzodiazepin-2-one
Figure imgf000089_0001
A mixture of 5-[2-chloro-6-(4-pyridin-4-ylpiperidin-l-yl)isonicotinoyl]-7,7- dimethyl-4,5,6,7-tetrahydro-lH-pyrazolo[4,3-c]pyridme (100 mg, 0.22 mmol), 3- piperidin-4-yl-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one trifluoroacetate (79 mg, 0.22 mmol), and potassium carbonate (91 mg, 0.66 mmol) in 1 ml of NMP is heated to 150 °C and stirred for 5 days in a sealed tube. Additional amounts of 3-piperidin-4-yl- l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one trifluoroacetate (60 mg) and potassium carbonate (91 mg) are added after days 1 and 3. The NMP is removed in vacuo, and the residue is purified by reverse phase HPLC to give the title compound as a brown solid. MS: 660.37 (M+H).
K. 3-(l-(4-(3,3-Dimethyl-4-oxopiperidine-l-carbonyl)pyridin-2-yl)piperidin-4- yl)-4,5-dihydro-lH-benzo[d][l,3]diazepin-2(3H)-one
Figure imgf000089_0002
A mixture of l-(2-chloroisonicotinoyl)-3,3-dimethylpiperidin-4-one (2.22 g, 8.34 mmol), 3-piperidin-4-yl-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one TFA salt (3 g, 8.34 mmol) and potassium carbonate (3.45 g, 25 mmol) in NMP (20 mL) is heated at 150 °C for 24 h. The reaction is cooled to rt, diluted with EtOAc, washed with water and brine, dried over Na2SO4, and concentrated under reduced pressure. Purification by silica gel chromatography (CH2Cl2 : THF = 6 : 1) affords the title compound as a yellow solid. 1H-NMR (400 MHz, CDCl3) δ : 8.23 (d, IH), 7.11 (t, IH), 7.04 (d, IH), 6.89 (t, IH), 6.68- 6.71 (m, 2H), 6.56 (d, IH), 6.47 (s, IH), 4.45-4.57 (m, IH), 4.02 (t, IH), 3.65-3.75 (m, 2H), 3.41-3.45 (m, 3H), 2.95-3.02 (m, 4H), 2.63 (t, IH), 2.48 (t, IH), 1.62-1.87 (m, 6H), 1.22 (s, 3H), 1.05 (s, 3H). MS: 476.31 (M+H). L. 3-(l-(4-(3,3-Dimethyl-4-(methylamino)piperidine-l-carbonyl)pyridin-2- yl)piperidin-4-yl)-4,5-dihydro-lH-benzo[d][l,3]diazepin-2(3H)-one
Figure imgf000090_0001
A mixture of methylamine HCl salt (42.4 mg, 0.63 mmol) and KOH (35.3 mg, 0.63 mmol) in MeOH (1 mL) in a sealed vial is stirred at 30 °C for 30 min. 3-(l-(4-(3,3- Dimethyl-4-oxopiperidine-l-carbonyl)pyridin-2-yl)piperidin-4-yl)-4,5-dihydro-lH- benzo[d][l,3]diazepin-2(3H)-one (30 mg, 0.063 mmol) is then added. The mixture is stirred at 30 °C for 6 h, and NaBH3CN is then added. The mixture is stirred at rt for 14 h. MeOH is removed under reduced pressure. The residue is partitioned between 50% saturated NaHCO3 and EtOAc, and the aqueous layer is extracted with EtOAc. The combined organic layer is washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The residue is purified by PTLC (CH2Cl2 : MeOH = 9 : 1) to afford the title compound as a colorless oil. 1H-NMR (400 MHz, CDCl3) δ: 8.19 (d, IH), 7.03-7.11 (m, 2H), 6.89 (m, IH), 6.70 (d, IH), 6.50-6.63 (m, 3H), 4.43-4.57 (m, 3.6H), 4.19 (m, 0.4H), 3.67 (m, 0.5H), 3.41-3.44 (m, 2H), 3.24 (m, 0.5H), 2.91-3.05 (m, 5H), 2.80 (d,
0.5H), 2.68 (d, 0.5H), 2.44 (s, 1.5H), 2.43 (s, 1.5H), 2.24 (m, IH), 1.65-1.98 (m, 6H), 1.36 (m, IH), 1.06 (s, 1.5H), 0.96 (s, 1.5H), 0.88 (s, 1.5H), 0.80 (s, 1.5H). MS: 491.35 (M+H). M. 3-(l-(4-(4-Amino-3,3-dimethylpiperidine-l-carbonyl)pyridin-2-yl)piperidin- 4-yl)-4,5-dihydro-lH-benzo[d][l,3]diazepin-2(3H)-one
Figure imgf000090_0002
Ammonium acetate (1.17 g, 15.2 mmol) is added to a stirred solution of 3-(l-(4- (3,3-dimethyl-4-oxopiperidine-l-carbonyl)pyridin-2-yl)piperidin-4-yl)-4,5-dihydro-lH- benzo[d][l,3]diazepin-2(3H)-one (0.72 g, 1.52 mmol) in MeOH (60 mL), and stirring is continued at rt overnight. NaBH3CN (114 mg, 1.82 mmol) is added, and the reaction is stirred at rt for 6 h. MeOH is removed under reduced pressure. The residue is partitioned between EtOAc and IN NaOH, and the EtOAc layer is washed with water and brine, dried over Na2SO4, and concentrated under reduced pressure. The residue is purified by silica gel chromatography (CH2Cl2 : MeOH = 95 : 5) to afford the title compound as a yellow foam. 1H-NMR (400 MHz, CDCl3) δ: 8.20(d, IH), 7.03-7.12 (m, 2H), 6.89 (m, IH), 6.69 (d, IH), 6.61 (m, IH), 6.51 (m, IH), 6.42 (s, IH), 4.44-4.56 (m, 3.5H), 4.24 (m, 0.5H), 3.64 (m, 0.5H), 3.41-3.44 (m, 2H), 3.30 (m, 0.5H), 2.62-3.07 (m, 7H), 1.40-1.89 (m, 8H), 1.04 (s, 1.5H), 0.94 (s, 1.5H), 0.86 (s, 1.5H), 0.79 (s, 1.5H). MS: 477.35 (M+H).
N. 3-(l-(4-(4-(2-Methoxyethylamino)-3,3-dimethylpiperidine-l-carbonyl)pyridin- 2-yl)piperidin-4-yl)-4,5-dihydro-lH-benzo[d][l,3]diazepin-2(3H)-one
Figure imgf000091_0001
A mixture of 3 -( 1 -(4-(3 ,3 -dimethyl-4-oxopiperidine- 1 -carbonyl)p yridin-2- yl)piperidin-4-yl)-4,5-dihydro-lH-benzo[d][l,3]diazepin-2(3H)-one (30 mg, 0.063 mmol), 2-methoxyethylamine (4.8 mg, 0.063 mmol), HOAc (5 μL) and NaBH(OAc)3 in DCE (2 mL) is stirred at 50 0C overnight. The reaction is cooled to rt, diluted with DCM, washed with IN NaOH and brine, dried over Na2SO4, and concentrated under reduced pressure. Purification by PTLC (CH2Cl2 : THF = 2 : 1) affords the title compound as a white solid. 1H-NMR (400 MHz, CDCl3) δ: 8.19(d, IH), 7.03-7.12 (m, 2H), 6.89 (m, IH), 6.69 (d,
IH), 6.61 (m, IH), 6.48-6.51 (m, 2H), 4.43-4.56 (m, 3.5H), 4.20 (m, 0.5H), 3.24-3.68 (m, 8H), 2.63-3.06 (m, 8H), 2.34 (m, IH), 1.50-1.89 (m, 7H), 1.06 (s, 1.5H), 0.97 (s, 1.5H), 0.88(s, 1.5H), 0.81(s, 1.5H). MS: 535.30 (M+H).
O. 3-(l-(5-(4-(2-Hydroxypropan-2-yl)piperidine-l-carbonyl)-4-methylthiazol-2- yl)piperidin-4-yl)-4,5-dihydro-lH-benzo[d][l,3]diazepin-2(3H)-one
Figure imgf000091_0002
To a solution of 4-methyl-2-(4-(2-oxo-l,2,4,5-tetrahydrobenzo[d][l,3]diazepin-3- yl)piperidin-l-yl)thiazole-5-carboxylic acid (19.3 mg, 0.05 mmol) in DMA (0.5 ml) is added 2-piperidin-4-yl-propan-2-ol (7.2 mg, 0.05 mmol), TEA (0.125 mL of IM solution in toluene, 0.125 mmol) and BOP (0.375 mL of 0.2 M solution in CH2Cl2, 0.075 mmol). The mixture is heated at 40 °C overnight. The reaction is cooled to rt, diluted with EtOAc, washed with IN NaOH, water and brine, dried over Na2SO4, and concentrated under reduced pressure. The residue is dissolved in 1 mL of EtOAc, and loaded evenly onto two SPE cartridges (0.5 g silica gel). Each cartridge is eluted with EtOAc (2 x 3 mL) to remove nonpolar impurities, then with CH2Cl2-MeOH (9/1, 5 mL). Concentration of the CH2Cl2-MeOH fractions affords the title compound as a yellow oil. 1H-NMR (400 MHz, CDCl3) δ: 7.11 (t, IH), 7.05 (d, IH), 6.90 (t, IH), 6.70 (d, IH), 6.48 (s, IH), 4.51 (m, IH), 4.35-4.40 (m, 2H), 4.05-4.13 (m, 2H), 3.44-3.47 (m, 2H), 3.09-3.17 (m, 2H), 2.96-3.00 (m, 2H), 2.81-2.87 (t, 2H), 2.26 (s, 3H), 1.74-1.89 (m, 6H), 1.54 (m, IH), 1.24-1.35 (m, 3H), 1.20 (s, 6H). MS: 512.22 (M+H).
P. l-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-methylpyridin-2-l}piperidin-4-yl)imidazolidine-2,4-dione
Figure imgf000092_0001
A mixture of 5-(2-chloro-6-methylisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro- lH-pyrazolo[4,3-c]pyridine (100 mg, 0.33 mmol), l-piperidin-4-ylimidazolidine-2,4- dione trifluoroacetate (196 mg, 0.66 mmol), and diisopropylethylamine (0.23 ml, 1.32 mmol) in 1.0 ml of NMP is heated to 150 0C and stirred for 16 h in a sealed tube. The NMP is removed in vacuo, and the residue is purified by reverse phase HPLC to give the title compound as a light brown solid. 1H NMR (400 MHz, DMSO-d6) δ 8.20 (IH, s), 7.40 (IH, s), 7.25 (IH, s), 6.56 (IH, s), 6.38 (IH, d, J 5.2), 4.60 (IH, s), 4.42 (2H, m), 4.30 (IH, s), 3.93 (IH, m), 3.89 (2H, s), 3.61 (IH, s), 3.27 (IH, s), 2.81 (2H, t, J 12.8), 2.30 (3H, d, J3.6), 1.69 (2H, m), 1.53 (2H, m), 1.24 (3H, s), 1.10 (3H, s). MS: 452.22 (M+H). Q. l-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-methylpyridin-2-l}piperidin-4-yl)-l,3-dihydro-2H-imidazo[4,5-b]pyridin- 2-one
Figure imgf000092_0002
A mixture of 5-(2-chloro-6-methylisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro- lH-pyrazolo[4,3-c]pyridine (150 mg, 0.49 mmol), l-piperidin-4-yl-l,3-dihydro-2H- imidazo[4,5-b]pyridin-2-one bis-hydrochloride (143 mg, 0.66 mmol), and diisopropylethylamine (0.34 ml, 1.96 mmol) in 1.0 ml of NMP is heated to 150 °C and stirred for 7 days in a sealed tube. Additional amounts (100 mg each) of l-piperidin-4-yl- l,3-dihydro-2H-imidazo[4,5-b]pyridin-2-one bis-hydrochloride are added after days 2, 4, and 6. The NMP is removed in vacuo, and the residue is purified by silica gel column chromatography (gradient from 50% EtOAc/CH2Cl2 to 5% MeOH/EtOAc) to afford the title compound as a light brown solid. 1H NMR (400 MHz, DMSOd6) δ 11.54 (IH, s), 7.87 (IH, d, J4.8), 7.48 (IH, m), 7.28 (IH, bs), 6.95 (IH, m), 6.62 (IH, s), 6.42 (IH, s), 4.47 (6H, m), 3.62 (IH, s), 2.90 (2H, m), 2.33 (3H, d, J4), 2.22 (2H, m), 1.77 (2H, d, J 10), 1.24 (3H, s), 1.12 (3H, s). MS: 487.23 (M+H).
R. 3-(l-{4-[(7,7-Dimethyl-l,4,5,7-tetrahydro-6H-pyrazolo[3,4-c]pyridin-6- yl)carbonyl]-l,3-thiazol-2-yl}piperidin-4-yl)-3,4-dihydroquinazolin-2(lH)-one
Figure imgf000093_0001
A mixture of 2- [4-(2-oxo- 1 ,4-dihydroquinazolin-3 (2H)-yl)piperidin- 1 -yl]- 1 ,3 - thiazole-4-carboxylic acid (100 mg, 0.279 mmol), 7,7-dimethyl-4,5,6,7-tetrahydro-lH- pyrazolo[4,3-c]pyridine dihydrochloride (52.4 mg, 0.279 mmol), and NEt3 (113 mg, 1.116 mmol) are dissolved in dichloromethane (2 mL). The mixture is cooled to 0 °C, 2-chloro- 1,3-dimethylimidazolinium chloride (71 mg, 0.419 mmol) is added, and the mixture is stirred at RT overnight. The mixture is concentrated in vacuo and purified by silica gel column chromatography to afford the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.35 (1 H, m), 7.21 (1 H, s), 7.17 (2 H, m), 7.05 (1 H, d), 6.95 (1 H, m), 6.70 (1 H, d), 4.88 (1 H, broad s), 4.77 (1 H, broad s), 4.64 (1 H, m), 4.33 (2 H, s), 4.14 (2 H, d), 3.92 (1 H, broad s), 3.77 (1 H, broad s), 3.20 (2 H, m), 1.94 (2 H, m), 1.83 (2 H, m), 1.39 (6 H, s). MS: 492.30 (M+).
S. 3-(l-{5-Chloro-3-[(7,7-dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3- c]pyridin-5-yl)carbonyl]isothiazol-4-yl}piperidin-4-yl)-3,4-dihydroquinazolin-2(lH)-one
Figure imgf000093_0002
5-[(4,5-Dichloroisothiazol-3-yl)carbonyl]-7,7-dimethyl-4,5,6,7-tetrahydro-lH- pyrazolo[4,3-c]pyridine (120 mg, 0.362 mmol), 3-piperidin-4-yl-3,4-dihydroquinazolin- 2(lH)-one trifluoroacetate (104 mg, 0.302 mmol), and K2CO3 (167 mg, 1.208 mmol) are added to NMP (1.5 mL) and the resulting mixture is heated at 150 °C overnight. DCM (30 mL) is added and the mixture is washed with water (5 ml x 3), brine (5 mL x 2), and dried over MgSO4. The solvent is removed in vacuo and silica gel column chromatography gives the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 7.46 (1 H, s), 7.38 (1 H, s), 7.19 (1 H, t, J 8), 7.09 (1 H, m), 6.96 (1 H, t, J 7.4), 6.71 (1 H, d, J 8), 4.85 (1 H, s), 4.93 (1 H, m), 4.45 (1 H, s), 4.38 (2 H, m), 4.02 (2 H, broad m), 3.80 (1 H, s), 3.44 (1 H, s), 3.08 (2 H, t), 2.07 (2 H, m), 1.84 (2 H, m), 1.41 (3 H, s), 1.27 (3 H, s). MS: 526.32 (M+). T. I1- {4-[(7,7-Dimethyl- 1 ,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]pyridin-2-yl}spiro[isoindole-l,4'-piperidin]-3(2H)-one
Figure imgf000094_0001
TFA (23.03 g, 0.202 mol) is added to 2-(3,5-dimethoxybenzyl)-l'-{4-[(7,7- dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)carbonyl]pyridin-2- yl}spiro[isoindole-l,4'-piperidin]-3(2H)-one (160 mg, 0.264 mmol), and the resulting solution is refluxed overnight. The volatiles are removed in vacuo. DCM (10 mL) is added and to the resulting solution is added saturated NaHCO3 solution (10 mL). This mixture is stirred for 10 min, the organic phase is separated and the aqueous phase is washed with DCM (20 mL x 3). The organic phases are combined and dried over MgSO4. The solvent is removed in vacuo and silica gel column chromatography affords the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 9.51 (1 H, broad s), 8.48 (1 H, broad s), 8.26 (1 H, d), 7.87 (1 H, d), 7.62 (1 H, m), 7.51 (1 H, m), 7.44 (2 H, m), 6.69 (1 H, s), 6.60 (1 H, m), 4.85 (1 H, s), 4.53 (2 H, m), 4.41 (1 H, s), 3.78 (1 H, s), 3.32 (1 H, s), 3.26 (2 H, m). MS: 457.31 (M+). U. l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]pyridin-2-yl}spiro[piperidine-4,7'-pyrrolo[3,4-b]pyridin]-5'(6Η)-one
Figure imgf000095_0001
TFA (23.03 g, 0.202 mol) is added to 6'-(3,5-dimethoxybenzyl)-l-{4-[(7,7- dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)carbonyl]pyridin-2- yl}spiro[piperidine-4,7'-pyiτolo[3,4-b]pyridin]-5'(6Η)-one (146 mg, 0.240 mmol), and the resulting solution is reflux ed overnight. The volatiles are removed in vacuo. DCM (100 mL) is added and to the resulting solution is added saturated NaHCO3 solution (10 mL). This mixture is stirred for 10 min, the organic phase is separated and the aqueous phase is washed with DCM (20 mL x 3). The organic phases are combined and dried over MgSO4. The solvent is removed in vacuo and silica gel column chromatography affords the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 9.17 (1 H, broad s), 8.75 (1 H, m), 8.26 (1 H, m), 8.15 (1 H, d, J 10), 7.43 (2 H, m), 7.24 (1 H, s), 6.71 (1 H, s), 6.60 (1 H, d), 4.84 (1 H, m), 4.48 (2 H, m), 3.78 (1 H, s), 3.48 (1 H, s), 3.44 (2 H, m), 3.34 (1 H, s), 2.40 (2 H, m), 1.73 (2 H, m), 1.39 (3 H, s), 1.13 (3 H, s). MS: 458.29 (M+).
V. 3-(l-{4-[(4-Pyridin-4-ylpiperidin-l-yl)carbonyl]pyridin-2-yl}piperidin-4-yl)- 3 ,4-dihydroquinazolin-2( 1 H)-one
Figure imgf000095_0002
2-Chloro-4-[(4-pyridin-4-ylpiperidin-l-yl)carbonyl]pyridine (100 mg, 0.331 mmol), 3-piperidin-4-yl-3,4-dihydroquinazolin-2(lH)-one trifluoroacetate (137.3 mg, 0.398 mmol), and K2CO3 (183 mg, 1.324 mmol) are added to DMA (1.0 mL) and the resulting mixture is heated at 150 °C overnight. DCM (30 mL) is added, and the mixture is washed with water (5 mL x 3) and brine (5 mL x 2), and dried over MgSO4. The solvent is removed in vacuo and silica gel column chromatography affords the title compound as a light yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.54 (2 H, m), 8.22 (1 H, d, J 6.4), 7.13-7.19 (3 H, m), 6.91-7.05 (3 H, m), 6.66-6.69 (2 H, m), 6.56 (1 H, m), 4.88 (1 H, m), 4.67 (1 H, m), 4.49 (2 H, d), 4.30 (2 H, s), 3.86 (1 H, d), 3.15 (1 H, m), 3.01 (2 H, m), 2.76-2.90 (2 H, m), 1.57-2.04 (8 H, m). MS: 497.35 (M+).
W. l'-{4-[(7,7-dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]pyridin-2-yl} spiro [indole-3 ,4'-piperidin] -2( 1 H)-one
Figure imgf000096_0001
To a dried three-necked flask bubbled with N2, ammonia (8 mL) is introduced with a cold finger. A mixture of 1 -benzyl- 1'- {4-[(7,7-dimethyl- 1, 4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5-yl)carbonyl]pyridin-2-yl}spiro[indole-3,4'-piperidin]-2(lH)-one (150 mg, 0.274 mmol) in THF (5 mL) is added and the resulting mixture is cooled to -78 °C. Lithium (11.4 mg, 1.65 mmol) is added and the resulting mixture is stirred at -78 °C for 30 min. EtOH (126 mg, 2.74 mmol) is added to quench the reaction and the reaction mixture is stirred at -78 0C to rt overnight. Water (10 mL) is added and the resulting mixture is extracted with DCM (20 mL x 5). The extract is dried over MgSO4, solvent is removed in vacuo and silica gel column chromatography affords the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.26 (1 H, m), 7.83 (1 H, s), 7.23-7.29 (3 H, m), 7.04 (1 H, m), 6.89 (1 H, m), 6.70 (1 H, s), 6.57 (1 H, m), (1 H, s). MS: 457.28 (M+).
X. 3-(l-{4-[(7,7-Dimethyl-l,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-methoxypyridin-2-yl}piperidin-4-yl)-l,3,4,5-tetrahydro-2H-l,3- benzodiazepin-2-one
Figure imgf000096_0002
5-(2-Chloro-6-methoxyisonicotinoyl)-7,7-dimethyl-4,5,6,7-tetrahydro-lH- pyrazolo[4,3-c]pyridine (71 mg, 0.221 mmol), 3-piperidin-4-yl-l,3,4,5-tetrahydro-2H-l,3- benzodiazepin-2-one trifluoroacetate (96 mg, 0.266 mmol), and K2CO3 (122 mg, 0.884 mmol) are added to DMA (1.0 mL) and the resulting mixture is heated at 150 °C overnight. The mixture is diluted with H2O (10 mL) and the resulting mixture is extracted with DCM (15 mL * 3). The extracts are dried over MgSO4, solvent is removed in vacuo and silica gel column chromatography affords the title compound as a light yellow solid. 1H NMR (400 MHz, CDCl3) δ 7.38 (1 H, s), 7.26 (1 H, s), 7.10 (1 H, m), 7.04 (1 H, m), 6.89 (1 H, m), 6.72 (1 H, d, J 8), 6.60 (1 H, s), 6.17 (1 H, d), 6.01 (1 H, m), 4.79 (1 H, s), 4.42-4.53 (3 H, m), 3.89 (2 H, m), 3.76 (1 H, s), 3.39-3.51 (3 H, m), 2.96 (3 H, m), 1.80- 1.83 (2 H, m), 1.72 (2 H, m), 1.39 (3 H, s), 1.25 (3 H, s). MS: 530.41 (M+).
Y. 9- {2-[4-(2-Oxo- 1 ,2,4,5-tetrahydro-3H- 1 ,3-benzodiazepin-3-yl)piperidin- 1 - yljisonicotinoyl} -9-azadispiro[2.1.2.3]decan-4-one
Figure imgf000097_0001
9-(2-Chloroisonicotinoyl)-9-azadispiro[2.1.2.3]decan-4-one (50 mg, 0.172 mol), 3- piperidin-4-yl-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one trifluoroacetate (74 mg, 0.206 mmol), and K2CO3 (122 mg, 0.884 mmol) are added to DMA (1.0 mL) and the resulting mixture is heated at 150 0C overnight. The reaction mixture is diluted with H2O (10 mL) and the resulting mixture is extracted with DCM (15 mL x 3). The extracts are dried over MgSO4, solvent is removed in vacuo and silica gel column chromatography affords the title compound as a white solid. 1H NMR (400 MHz, CDCl3) δ 8.22 (1 H, d, J 5), 7.26 (1 H, s), 7.10 (2 H, m), 7.04 (1 H, m), 6.89 (1 H, m), 6.70 (2 H, m), 6.57 (1 H, m), 6.50 (1 H, s), 4.55 -4.57 (3 H, m), 3.94 (2 H, s), 3.62 (2 H, s), 3.43 (2 H, m), 2.96-3.01 (4 H, m), 1.84-1.86 (2 H, m), 1.65-1.75 (2 H, m), 1.38 (2 H, m), 1.31 (2 H, m), 1.00 (2 H, m), 1.65 (2 H, m). MS: 500.31 (M+).
Z. 3-{l-[4-(l',4'-Dihydrospiro[cyclopropane-l,7l-pyrazolo[4,3-c]pyridin]-5l(6Η)- ylcarbonyl)pyridin-2-yl]piperidin-4-yl}-l,3,4,5-tetrahydro-2H-l,3-benzodiazepin-2-one
Figure imgf000097_0002
5'-(2-Chloroisonicotinoyl)-r,4',5',6'-tetrahydrospiro[cyclopropane-l,7'- pyrazolo[4,3-c]pyridine] (70 mg, 0.242 mmol), 3-piperidin-4-yl-l,3,4,5-tetrahydro-2H- l,3-benzodiazepin-2-one trifluoroacetate (104.5 mg, 0.291 mmol), and K2CO3 (167 mg, 1.21 mmol) are added to DMA (1.0 mL) and the resulting mixture is heated at 150 °C overnight. The reaction mixture is diluted with H2O (10 mL) and the resulting mixture is extracted with DCM (15 mL * 3). The extracts are dried over MgSO4 and solvent is removed in vacuo. Silica gel column chromatography affords the title compound as a yellow solid. 1H NMR (400 MHz, CDCl3) δ 8.23 (1 H, d, J 5), 7.40 (1 H, s), 7.23 (1 H, s), 7.11 (1 H, m), 7.05 (1 H, m), 6.90 (1 H, s), 6.72 (1 H, m), 6.69 (1 H, s), 6.64 (1 H, s), 6.57 (1 H, m), 4.90 (1 H, s), 4.55 (2 H, s), 4.47 (2 H, m), 3.88 (1 H, s), 3.42-3.49 (3 H, m), 2.96-3.02 (4 H, m), 1.84 (2 H, m), 1.66-1.75 (2 H, m), 1.12-1.28 (4 H, m). MS: 498.35 (M+).
EXAMPLE 3. ADDITIONAL REPRESENTATIVE AMIDE-SUBSTITUTED ARYL PIPERIDINES
Using routine modifications, the starting materials may be varied and additional steps employed to produce other compounds provided herein. Compounds listed in Table I are prepared using such methods. All compounds provided in Table I exhibit an IC5O (determined as described in Example 5) that is 10 micromolar or less; compounds with a "*" in the column headed "IC50" exhibit an IC5O (determined as described in Example 5) that is 2 micromolar or less. Mass spectroscopy data is provided as M+l in the column headed "MS," with the retention times ("Ret Time") given in minutes.
Table I: REPRESENTATIVE AMIDE-SUBSTITUTED ARYL PIPERIDINES
Ret
COMPOUND NAME MS Time IC42
359.22 1.02
436.20 1.02
Figure imgf000098_0001
458.17 1.04
Figure imgf000098_0002
y7 Ret
COMPOUND NAME MS Time IC42
,..,, . ._ 50932 1/l2
432.24 0.97
486.32 1.11 ,4- 1 H)-
462.31 1.11
- ,4- 464.27 1.1
Figure imgf000099_0001
Ret MS Time IC50 ,3-
594.08 1.19 5-
Figure imgf000100_0001
505.26 1.01
500.31 1.12 ,4,5-
518.28 1.24 ,4,5-
Figure imgf000100_0002
500.31 1.11
Figure imgf000100_0003
Ret
COMPOUND NAME MS Time IC5Q
3-(l-{4-[(4-pyridin-4- ylpiperidin-1- yl)carbonyl]pyridin-2- y 1} piperidin-4-y I)- 1 ,3 ,4,5- 511.51 1.05 tetrahydro-2H-l,3-
Figure imgf000101_0001
benzodiazepin-2-one
- ,4,5- 463.48 1.04
-
449.45 1
550.26 1.15 1,3,4,5-
-
536.26 1.14 ,4- 1 H)-
Figure imgf000101_0002
3-(l-{4-[(4-amino-3,3- dimethylpiperidin- 1 - yl)carbonyl]pyridin-2- y 1 } piperidin-4-yl)- 1 ,3 ,4,5- 0.99 tetrahydro-2H-l,3- benzodiazepin-2-one
Figure imgf000101_0003
598.39 1.1
Figure imgf000101_0004
Ret
COMPOUND NAME MS Time ICio
534.25 1.24
1 -
478.29 1.06 H)-
,4- 448.26 1.13 1 H)-
464.25 1.08
500.29 1.23 , 3,4,5-
500.47 1.1
Figure imgf000102_0001
486.48 1.08 1 H)-
Figure imgf000102_0002
Ret
COMPOUND NAME MS Time IC≤o
3-(l-{4-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl] - 1 ,3 -thiazol- 506.24 1.23 2-yl}piperidin-4-yl)- l,3,4,5-tetrahydro-2H-l,3-
Figure imgf000103_0001
benzodiazepin-2-one
492.24 1.22 ,4- 1 H)-
Figure imgf000103_0002
N,5-dimethyl-N-[2- methyl-2-( 1 H-pyrazol-5- yl)propyl]-2-[4-(2-oxo- W. 1 ,2,4,5-tetrahydro-3H- 1 ,3- 1.22 benzodiazepin-3- 522.27
H b yl)piperidin- 1 -yl] -1,3-
Figure imgf000103_0003
thiazole-4-carboxamide
3-(l-{5-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl] -4-methyl- l,3-thiazol-2-yl}piperidin- 520.44 1.2
4-yl)-l,3,4,5-tetrahydro-
2H- 1 ,3-benzodiazepin-2- one
3-(l-{5-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-l,3-thiazol- 505 20 1 22
2-yl} piperidin-4-yl)- l,3,4,5-tetrahydro-2H-l,3-
Figure imgf000103_0004
benzodiazepin-2-one
2-methyl-N-[l-({4- methyl-2-[4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- benzodiazepin-3- y l)piperidin- 1 -yl] - 1 , 3 - 539.23 1.21 thiazol-5-
Figure imgf000103_0005
yl} carbonyl)piperidin-4- yl]propanamide l-(l-{5-[(7,7-dimethyl-
518.23 1.2
Figure imgf000103_0006
Ret
COMPOUND NAME MS Time IC™
3-(l-{5-[(3-hydroxy-3- methylpyrrolidin- 1 - yl)carbonyl]-4-methyl- l,3-thiazol-2-yl}piperidin- 470.26 1 17 4-yl)-l ,3,4,5-tetrahydro-
Figure imgf000104_0001
2H-l,3-benzodiazepin-2- one
3-(l-{5-[(3-hydroxy-3- methylpiperidin- 1 - yl)carbonyl] -4-methyl- l,3-thiazol-2-yl}piperidin- 484 27 1 19
4-yl)-l ,3,4,5-tetrahydτo-
Figure imgf000104_0002
2H- 1 ,3-benzodiazepin-2- one
N-(2-hydroxy-2- methylpropyl)-N,4- dimethyl-2- [4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- 472.24 1.1 1 benzodiazepin-3-
Figure imgf000104_0003
y l)piperidin- 1 -yl] - 1 , 3 - thiazole-5-carboxamide
1 - 442 21 1 23
Figure imgf000104_0004
3-[ 1 -(5- {[3-(l -hydroxy- 1- methylethyl)piperidin- 1 - yljcarbonyl} -4-methyl- l,3-thiazol-2-yl)piperidin- 5-12 29 1 23 4-yl] - 1 ,3 ,4, 5 -tetrahydro-
Figure imgf000104_0005
2H- 1 ,3-benzodiazepin-2- one
4g2 30 -| 32
Figure imgf000104_0006
ethyl 4-( {4-methyl-2-[4-
(2-oxo- 1, 2,4,5-tetrahydro-
3H-l,3-benzodiazepin-3- yl)piperidin-l-yl]-l,3- 527.26 1.23 thiazol-5-
Figure imgf000104_0007
yl} carbonyl)piperazine- 1 - carboxylate Ret
COMPOUND NAME MS Time IC50 ethyl l-({4-methyl-2-[4- (2-oxo- 1, 2,4,5-tetrahydro- 3H-l,3-benzodiazepin-3- yl)piperidin-l-yl]-l,3- 526.26 1.25 thiazol-5- yl} carbonyl)piperidine-4- carboxylate ethyl l-({4-methyl-2-[4-
(2-oxo- 1, 2,4,5-tetrahydro-
3H-l,3-benzodiazepin-3- yl)piperidin-l-yl]-l,3- 526.26 1 .25 thiazol-5-
Figure imgf000105_0001
yl} carbonyl)piperidine-3- carboxylate
3-(l-{5-[(4-
497 26 1.16
Figure imgf000105_0002
N-[l-({4-methyl-2-[4-(2- 0x0- 1 ,2,4,5-tetrahydro- 3H-l,3-benzodiazepin-3- yl)piperidin-l-yl]-l,3- 497.24 1.09 thiazol-5- yl}carbonyl)pyrrolidin-3- yl]acetamide
3-(l-{5-[(4-acetyl-l,4- diazepan- 1 -yl)carbonyl]-
4-methyl- 1 ,3-thiazol-2- yl}piperidin-4-yl)-l,3,4,5- 511.26 1 .15 tetrahydro-2H-l,3- benzodiazepin-2-one
N,N-diethyl-l-({4- methyl-2-[4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- benzodiazepin-3- yl)piperidin- 1 -yl]- 1,3- 553.30 1.23 thiazol-5-
Figure imgf000105_0003
yl} carbonyl)piperidine-3- carboxamide
434 21 \ 23
Figure imgf000105_0004
Ret
COMPOUND NAME MS Time IC5
484.26 1.24
Figure imgf000106_0001
l-
486.26 1.17 1 ,3-
Figure imgf000106_0002
486.24 1.19 ,3 -
Figure imgf000106_0003
N-(2-methoxyethyl)-N,4- dimethyl-2-[4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- benzodiazepin-3 - 458.25 1.19 yl)piperidin- 1 -yl]- 1 ,3 - thiazole-5 -carboxamide
N-(2-methoxyethyl)-4- methyl-2-[4-(2-oxo-
1 ,2,4,5-tetrahydro-3H- 1,3- benzodiazepin-3- 486.25 1.18 yl)piperidin- 1 -yl]-N- propyl- 1 ,3-thiazole-5-
Figure imgf000106_0004
carboxamide l-
472.28 1.21
Figure imgf000106_0005
Figure imgf000106_0006
Ret
COMPOUND NAME MS Time i IjC-se ethyl N-methyl-N-({4- methyl-2-[4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- benzodiazepin-3- y l)piperidin- 1 -yl] - 1 , 3 - thiazol-5-yl}carbonyl)- beta-alaninate methyl N-methyl-N-({4- methyl-2-[4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- benzodiazepin-3- 472.24 1 .19 yl)piperidin- 1 -yl]- 1 ,3- thiazol-5- yl} carbonyl)glycinate methyl l-({4-methyl-2-[4-
(2-oxo- 1 ,2,4,5-tetrahydro-
3H-l,3-benzodiazepin-3- yl)piperidin- 1 -yl]- 1 ,3- thiazol-5- yl} carbonyl)prolinate
Figure imgf000107_0001
3-(l-{5-[(4-
434 27 1.21
Figure imgf000107_0002
3-[l-(5-{[(3S)-3- methoxypyrrolidin- 1 - yl]carbonyl}-4-methyl-
1 ,3-thiazol-2-yl)piperidin- 470.25 1.2
4-yl]-l,3,4,5-tetrahydro-
Figure imgf000107_0003
2H-l,3-benzodiazepin-2- one
3-(l-{5-[(3-
O methoxypiperidin- 1 - yl)carbonyl] -4-methyl-
N-/ N-{ J Y l,3-thiazol-2-yl}piperidin- 434 27 1.21
4-yl)- 1 ,3,4,5-tetrahydro-
2H- 1 ,3-benzodiazepin-2- one
3-[l-(5-{[3-
(methoxymethyl)piperidin - 1 -yl]carbonyl} -4-methyl- 1 ,3-thiazol-2-yl)piperidin- 498.27 1.24 4-yl]-l,3,4,5-tetrahydro-
Figure imgf000107_0004
2H- 1 ,3-benzodiazepin-2- one
1 -
512.28 1.25
Figure imgf000107_0005
Ret
COMPOU]NJD NAME MS Time IC42 one
498.28 1.24
498.27 1.24
Figure imgf000108_0001
Figure imgf000108_0002
one
N,4-dimethyl-2-[4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- benzodiazepin-3- yl)piperidin- 1 -yl] -N- 470.25 1.18 I (tetrahydrofuran-3-yl)-
Figure imgf000108_0003
l,3-thiazole-5- carboxamide
456.24 1.18
Figure imgf000108_0004
N-[l-({4-methyl-2-[4-(2-
OXO- 1 ,2,4,5-tetrahydro-
3H-l,3-benzodiazepin-3- y l)piperidin- 1 -yl] - 1 , 3 - 525.28 1.11 thiazol-5- yl} carbonyl)piperidin-4-
Figure imgf000108_0005
yljpropanamide Ret
COMPOUND NAME MS Time IC52
N-[l-({4-methyl-2-[4-(2- 0X0- 1 ,2,4,5-tetrahydro- 3H- 1 ,3-benzodiazepin-3-
72 yl)piperidin- 1 -yl]- 1 ,3- 511.26 1.09 thiazol-5- yl} carbonyl)piperidin-4-
Figure imgf000109_0001
yl]acetamide
73 560.21 1.29
74 560.22 1.29
75 546 2Q 1 25
76 490.22 1.28
Figure imgf000109_0002
N,4-dimethyl-N-(4- methylbenzyl)-2-[4-(2- 0X0- 1 ,2,4,5-tetrahydro-
77 3H-1 ,3-benzodiazepin-3- 504.22 1.31 yl)piperidin- 1 -yl]- 1 ,3- thiazole-5 -carboxamide
N,4-dimethyl-N-(3- methylbenzyl)-2-[4-(2- 0X0- 1 ,2,4,5-tetrahydro-
78 3H-l,3-benzodiazepin-3- 504.22 1.32 yl)piperidin- 1 -yl] - 1 ,3 -
Figure imgf000109_0003
thiazole-5-carboxamide
N-(4-fluorobenzyl)-N,4- dimethyl-2- [4-(2-oxo- N 1 ,2,4,5-tetrahydro-3H- 1 ,3-
79 ( benzodiazepin-3- 508.19 1.28
Figure imgf000109_0004
yl)piperidin- 1 -yl] - 1 ,3-
H 0 ■ CX1? thiazole-5-carboxamide Ret
COMPOUND NAME MS Time IC50
N-(3 -fluorobenzyl)-N,4- dimethyl-2-[4-(2-oxo- l,2,4,5-tetrahydro-3H-l,3- benzodiazeρin-3- 508.20 1 .28 yl)piperidin-l-yl]-l,3-
Figure imgf000110_0001
thiazole-5 -carboxamide
81 497.15 1 .2 ,5-
82 498.14 1 .33 ,5-
Figure imgf000110_0002
3-[ 1.(4- {[4-(l -hydroxy- 1- methylethyl)piperidin- 1 - yl]carbonyl } pyridin-2-
83 yl)piperidin-4-yl]-l,3,4,5- 492-21 1 -13 tetrahydro-2H-l,3-
Figure imgf000110_0003
benzodiazepin-2-one
84 507.21 1.29 ,3,4,5-
Figure imgf000110_0004
cis-3-[l-(4-{[(2R,6S)-2,6- dimethylmorpholin-4- yl]carbonyl}-6-
85 methylpyridin-2- 478.21 1 .15 yl)piperidin-4-yl]-l,3,4,5- tetrahydro-2H-l,3- benzodiazepin-2-one ciS-l-[l-(4-{[(2R,6S)-2,6- dimethylmorpholin-4- yl]carbonyl}pyridin-2-
86 yl)piperidin-4-yl]-l,3- 437.18 1.04 dihydro-2H-imidazo[4,5- b]pyridin-2-one
Figure imgf000110_0005
Ret
COMPOUND NAME MS Time IC™ cis-3-{ l-[4-{[(2R,6S)-2,6- dimethyltnoφholin-4- yl]carbonyl}-6-
(methylamino)pyridin-2- 493 22 1 16 yl]piperidin-4-yl } - 1 ,3,4,5- tetrahydro-2H-l,3- benzodiazepin-2-one cis-l-[l-(4-{[(2R,6S)-2,6- dimethylmoφholin-4- y ljcarbonyl} pyridin-2- yl)piperidin-4-yl]-4- 462.18 1.15 phenyl- 1 ,3-dihydro-2H- imidazol-2-one
3 -( 1 - {6-(dimethy lamino)- 4-[(7,7-dimethyl- 1,4,6,7- tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]pyridin-2- yl} piperidin-4-yl)- 1,3,4,5- tetrahydro-2H-l,3- benzodiazepin-2-one l-(l-{4-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-
(methoxymethyl)pyridin- 517.17 1.16
2-yl}piperidin-4-yl)- 1 ,3- dihydro-2H-imidazo[4,5-
Figure imgf000111_0001
b]pyridin-2-one
1 1 7
Figure imgf000111_0002
l-{4-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6- 502.17 1.25 methoxypyridin-2-yl}-
1 'H-spiro[piperidine-4,4'-
Figure imgf000111_0003
quinazolin]-2'(3'H)-one ICio
Figure imgf000112_0001
l-(l-{4-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6- methylpyridin-2- 542.19 1.15 y 1 } piperidin-4 -yl)-4 -(3 - methoxyphenyl)- 1 ,3-
Figure imgf000112_0002
dihydro-2H-imidazol-2- one -
528.17 1.14
Figure imgf000112_0003
3-(l-{5-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-4- (methoxymethyl)- 1 ,3- 550.16 1.6 thiazol-2-yl}piperidin-4- yl)-l,3,4,5-tetrahydro-2H-
Figure imgf000112_0004
1 ,3-benzodiazepin-2-one
3-(l-{5-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-4- (hydroxymethyl)-l,3- 536.21 1.26 thiazol-2-yl}piperidin-4- yl)- 1 ,3,4,5-tetrahydro-2H- 1 ,3-benzodiazepin-2-one
3-(l-{4-[(3-isopropyl-5,6- dihydro[l,2,4]triazolo[4,3
-a]pyrazin-7(8H)- yl)carbonyl]pyridin-2- 515.32 1.46 yl}piperidin-4-yl)- 1,3,4,5- tetrahydro-2H-l,3-
Figure imgf000112_0005
benzodiazepin-2-one Ret
COMPOUND NAME MS Time IC42 l-(l-{4-[(3-isopropyl-5,6-
Figure imgf000113_0001
one
2-(l-{4-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-
102 methylpyridin-2- 513.3 1.56 yl}piperidin-4-yl)-5- phenyl-2,4-dihydro-3H- l,2,4-triazol-3-one cis-2-[l-(4-{[(2R,6S)-2,6- dimethylmorpholin-4- yl]carbonyl}pyridin-2-
103 yl)piperidin-4-yl]-5- 463.28 1.58 phenyl-2,4-dihydro-3H- l,2,4-triazol-3-one
Figure imgf000113_0002
-
104 578.3 1.56
Figure imgf000113_0003
l-(l-{4-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5-
105 yl)carbonyl]pyridin-2- 473.23 1.47 yl}piperidin-4-yl)-l,3- dihydro-2H-imidazo[4,5- b]pyridin-2-one
4-(l-{4-[(7,7-dimethyl- l,4,6,7-tetrahydro-5H- pyrazolo[4,3-c]pyridin-5- yl)carbonyl]-6-
106 methylpyridin-2- 512.29 1.61 yl}piperidin-4-yl)-l- phenyl- 1 ,2-dihydro-3H-
Figure imgf000113_0004
pyrazol-3-one
107 - 484.28 2.18
Figure imgf000113_0005
Ret
COMPOUND NAME MS Time IC4O
108 512.3 2.26
109 497.29 2.71
-
1 10 528.18 1.78
Figure imgf000114_0001
3-(l-{4-[(8,8-dimethyl- 7,8-dihydropyrido[4,3- d]pyrimidin-6(5H)- yl)carbonyl]pyridin-2- 512.17 1.67 y 1l}1 pi *peri 'd A*i'~n*.-4 λ- .y Λl\)- Λl,3 *},4 A,5 C- tetrahydro-2H-l,3-
Figure imgf000114_0002
benzodiazepin-2-one
- 540.49 1.57
Figure imgf000114_0003
EXAMPLE 4. PREPARATION OF CGRP RECEPTOR-TRANSFECTED CELLS
This Example illustrates the preparation of CGRP receptor-transfected cells for use in CGRP receptor calcium mobilization assays (Example 5).
Complementary DNAs encoding the human receptor activity-modify protein 1 (hRAMP-1; GenBank #NM_005855) and human calcitonin receptor-like receptor (hCRLR; GenBank #NM_005795) are cloned in the pIRES mammalian expression vector (Clontech, Mountain View, CA). The hCRLR is subcloned into the plasmid 5' of the IRES element and the hRAMP-1 is subcloned into the plasmid 3' of the IRES element. This plasmid is transfected into HEK293 cells that stably express recombinant rat olfactory cyclic nucleotide gated channel subunit 1 (GenBank # X55519) and rat olfactory cyclic nucleotide gated channel subunit 2 (GenBank #U12623), thus generating cells that take up extracellular calcium via the rat olfactory cyclic nucleotide gated channel in response to increased intracellular cyclic AMP. A clonal cell line stably expressing human CGRP receptor is selected by neomycin resistance and by a calcium mobilization assay using calcium sensitive fluorescent dye as detailed below.
EXAMPLE 5. CALCIUM MOBILIZATION ASSAY This Example illustrates representative calcium mobilization assays for use in evaluating test compounds for agonist and antagonist activity at CGRP receptor.
The day prior to the experiment, cells expressing CGRP receptor are plated at a density of 25,000 cells/well in 384 well PDL coated BD Falcon plates. On the day of the experiment, the media is removed and 25 μL of 2.3 μM Fluo-4 AM dye (TEF Labs, Inc., Austin, TX) in KRH buffer (5 mM KCl, 115 raM NaCl, 960 μM MgH2PO4, 1 mM
MgSO4, 2 mM CaCl2, 5 mM glucose, 25 mM HEPES, 1 mM probenecid) is added. Plates are incubated at 37 °C, 5% CO2 for 1 hour. Wells are then washed with 50 μL KRH buffer. Wash buffer is removed and 25 μL fresh KRH is added to each well. Plates are incubated for 1 hour at room temperature, after which 25 μL of KRH and test compound (10OX final concentration) in DMSO or KRH and DMSO is transferred into the appropriate wells. Test compounds are tested in serial dilutions from 10 μM, 4 μM, 1 μM, or 0.1 μM stocks; for positive control cells, no test compound is added. The plates are incubated for an additional 1 hour at room temperature, and then transferred to a FLIPR384 instrument (Molecular Devices, Sunnyvale, CA) for calcium mobilization analysis.
For determination of antagonist activity, activating agonist (25 μL of KRH containing CGRP) is transferred using the FLIPR into the plate, such that the final CGRP concentration is the EC50 for the CGRP-mediated response in these cells (1.6 nM CGRP). Pn negative control cells, 25 μL of KRH without CGRP is added at this stage. Fluorescence is monitored for at least 4 minutes. The data is analyzed as follows. First, the average maximum relative fluorescent unit (RFU) response from the negative control wells (no CGRP) is subtracted from the maximum response detected for each of the other experimental wells. Second, average maximum RFU response is calculated for the agonist response to CGRP in the positive control wells (agonist wells). Then, percent inhibition for each compound tested is calculated using the equation:
0/ . , ., ... r i (agonist RFU response + compound RFU response) , 1 Λ_
% inhibition = [1- ^-5 ; . , „„ . 1T κ L ] x 100
(agonist RFU response)
The % inhibition data is plotted as a function of test compound concentration and test compound IC50 is determined using KALEID AGRAPH software (Synergy Software, Reading, PA) best fit of the data to the equation: y = m1*(l/(l+(m2/m0)m3)) where y is the percent inhibition, m0 is the concentration of the agonist (CGRP), mi is the maximum RFU, m2 corresponds to the test compound IC50 (the concentration required to provide a 50% decrease, relative to the response observed in the presence of CGRP and without antagonist) and m3 is the Hill coefficient. Antagonists of the CGRP receptor decrease the response induced by CGRP alone (in the absence of test compound) by at least about 20%, preferably by at least about 50%, and most preferably by at least 80%, at a concentration of 10 micromolar or less, preferably 4 micromolar or less, and more preferably 1 micromolar or less. The IC50 such antagonists is preferably below 1 micromolar, 100 nanomolar, 10 nanomolar or 1 nanomolar.
Similar assays are performed in the absence of added CGRP for the determination of agonist activity of the test compounds. Within such assays, the ability of a test compound to act as an agonist of CGRP receptor is determined by measuring the fluorescence response elicited by the test compound as a function of compound concentration. CGRP receptor antagonists that exhibit no detectable agonist activity elicit no detectable fluorescence at concentrations of 100 nanomolar, or 1, 4 or 10 micromolar.
Example 6. MDCK CYTOTOXICITY ASSAY This Example illustrates the evaluation of compound toxicity using a Madin Darby canine kidney (MDCK) cell cytotoxicity assay.
1 μL of test compound is added to each well of a clear bottom 96-well plate (PACKARD, Meriden, CT) to give final concentration of compound in the assay of 10 μM, 100 μM or 200 μM. Solvent without test compound is added to control wells. MDCK cells, ATCC no. CCL-34 (American Type Culture Collection, Manassas,
VA), are maintained in sterile conditions following the instructions in the ATCC production information sheet. Confluent MDCK cells are trypsinized, harvested, and diluted to a concentration of 0.1 x 106 cells/mL with warm (37°C) medium (VITACELL Minimum Essential Medium Eagle, ATCC catalog # 30-2003). 100 μL of diluted cells is added to each well, except for five standard curve control wells that contain 100 μL of warm medium without cells. The plate is then incubated at 370C under 95% O2, 5% CO2 for 2 hours with constant shaking. After incubation, 50 μL of mammalian cell lysis solution (from the Packard ATP-LITE-M Luminescent ATP detection kit) is added per well, the wells are covered with PACKARD TOPSEAL stickers, and plates are shaken at approximately 700 rpm on a suitable shaker for 2 min.
Compounds causing toxicity will decrease ATP production, relative to untreated cells. The ATP-LITE-M Luminescent ATP detection kit is generally used according to the manufacturer's instructions to measure ATP production in treated and untreated MDCK cells. PACKARD ATP LITE-M reagents are allowed to equilibrate to room temperature. Once equilibrated, the lyophilized substrate solution is reconstituted in 5.5 mL of substrate buffer solution (from kit). Lyophilized ATP standard solution is reconstituted in deionized water to give a 10 mM stock. For the five control wells, 10 μL of serially diluted PACKARD standard is added to each of the standard curve control wells to yield a final concentration in each subsequent well of 200 nM, 100 nM, 50 nM, 25 nM, and 12.5 nM. PACKARD substrate solution (50 μL) is added to all wells, which are then covered, and the plates are shaken at approximately 700 rpm on a suitable shaker for 2 min. A white PACKARD sticker is attached to the bottom of each plate and samples are dark adapted by wrapping plates in foil and placing in the dark for 10 min. Luminescence is then measured at 22°C using a luminescence counter (e.g., PACKARD TOPCOUNT Microplate Scintillation and Luminescence Counter or TECAN SPECTRAFLUOR PLUS), and ATP levels calculated from the standard curve. ATP levels in cells treated with test compound(s) are compared to the levels determined for untreated cells. Cells treated with 10 μM of a preferred test compound exhibit ATP levels that are at least 80%, preferably at least 90%, of the untreated cells. When a 100 μM concentration of the test compound is used, cells treated with preferred test compounds exhibit ATP levels that are at least 50%, preferably at least 80%, of the ATP levels detected in untreated cells.

Claims

What is claimed is:
1. A compound of formula:
Figure imgf000118_0001
or a pharmaceutically acceptable salt or hydrate thereof, wherein:
is a 5- or 6-membered heterocyclic ring, wherein X is C, CH, or N, such that the heterocyclic ring that:
(a) comprises at least one ring nitrogen atom;
(b) is substituted with oxo;
(c) is substituted with 0-2 Ci-C4alkyl; and
(d) is:
(i) substituted with a phenyl or 5- or 6-membered heteroaryl substituent; or (ii) fused to a phenyl or 5- or 6-membered heterocyclic ring; wherein each of which (i) and (ii) is substituted with from 0 to 4 substituents independently chosen from
(1) halogen, hydroxy, cyano, amino, and oxo; and
(2) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, C,-C6hydroxyalkyl, Ci-C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8cycloalkyl)C0-C4alkyl, mono- or di- (Ci-C6alkyl)aminoC0-C4alkyl, and (4- to 10-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino, aminocarbonyl, aminosulfonyl, Ci-C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-C6alkanoyl, Ci-C6alkylsulfonyl, Ci- C6alkylsulfinyl, Ci-Cόalkoxycarbonyl, mono- or di-(Ci-C6alkyl)aminocarbonyl, and mono- or di-(Ci-C6alkyl)aminosulfonyl;
Ar is a 5- or 6-membered heteroaryl that is substituted with from 0 to 2 substituents independently chosen from R3;
Ri is C]-C8aminoalkyl, Ci-C8hydroxyalkyl, C2-C8alkyl ether, phenylCo-Cόalkyl, or (4- to 12-membered heterocycle)C0-C6alkyl, each of which is substituted with from 0 to 6 substituents independently chosen from:
(i) halogen, hydroxy, cyano, amino, aminocarbonyl, aminosulfonyl and oxo; and (ii) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, Ci- C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8carbocycle)C0-C4alkyl, mono- or di- (Ci- C6alkyl)aminoC0-C4alkyl, and (4- to 12-membered heterocycle)Co-C4alkyl, each of which is substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino and Ci-C4alkyl; and
R2 is hydrogen, Ci-C6alkyl or Ci-C6haloalkyl; or Ri and R2 are taken together to form a 4- to 10-membered heterocycle that is substituted with from 0 to 6 substituents independently chosen from:
(i) halogen, hydroxy, cyano, amino, aminocarbonyl, aminosulfonyl and oxo; and
(ii) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, C2- C6alkyl ether, Ci-C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8cycloalkyl)C0-C4alkyl, mono- or di- (Ci-Cόalkyl)aminoCo-C4alkyl, phenylCo-C4alkyl, (4- to 12-membered heterocycle)C0-C4alkyl, and spiro C3-C8cycloalkyl; each of which is substituted with from 0 to 6 substituents independently chosen from hydroxy, oxo, halogen, amino, aminocarbonyl, aminosulfonyl, Q- Qalkyl, Ci-C6alkoxy, Ci-C6alkylthio, CrC6alkanoyl, Ci-C6alkylsulfonyl, Ci-C6alkylsulfmyl, Ci-C6alkoxycarbonyl, mono- or di-(Ci-C6alkyl)aminocarbonyl, and mono- or di-(Ci- C6alkyl)aminosulfonyl;
R3 represents from 0 to 3 substituents independently chosen from:
(i) halogen, hydroxy, cyano and amino; and
(ii) Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, and (4- to 7-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, Ci-C4alkyl, (C3-C8carbocycle)C0-C4alkyl and (4- to 10-membered heterocycle)Co-C4alkyl; or two substituents represented by R3 are taken together to form a fused 5- to 7-membered carbocycle or heterocycle that is substituted with from 0 to 3 substituents independently chosen from halogen, hydroxy, amino, Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci- C6haloalkyl, Ci-C6alkoxy, Ci-C6alkyl ether, mono- or di-(Ci-Cόalkyl)aminoCo-C2alkyl, and (4- to 7-membered heterocycle)Co-C4alkyl; and
R6 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, amino, cyano, Ci-C6alkyl, C]-C6haloalkyl, Ci-C6alkoxy and C2-C6alkyl ether.
2. The compound or salt or hydrate thereof according to claim 1 wherein: Ar is B-D K 3 wherein A, B, D and E are each CH, C, or N, such that exactly one or two of A, B, D and E are N;
Figure imgf000120_0001
X is N;
Z is C, CH, CH2, N or NH; ^^ represents a single or double bond; n is 0, 1 or 2; R4 Is: (i) oxo;
(ii) a phenyl or 5- or 6-membered heteroaryl substituent; or (iii) a fused phenyl or 5- or 6-membered heterocyclic ring; each of which (ii) and (iii) is substituted with from 0 to 4 substituents independently chosen from:
(a) halogen, hydroxy, cyano, amino and oxo; and
(b) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, C1- C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8cycloalkyl)C0-C4alkyl, mono- or di- (C]- C6alkyl)aminoCo-C4alkyl, and (4- to 10-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino, aminocarbonyl, aminosulfonyl, Ci-C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-C6alkanoyl, Ci-C6alkylsulfonyl, Ci-C6alkylsulfmyl, Ci-C6alkoxycarbonyl, mono- or CU-(C1- C6alkyl)aminocarbonyl, and mono- or di-(C]-C6alkyl)aminosulfonyl; and
R5 represents from 0 to 4 substituents independently chosen from Ci-C6alkyl, C2- C6alkenyl, C2-C6alkynyl, and (C3-C8cycloalkyl)C0-C4alkyl.
3. The compound or salt or hydrate thereof according to claim 2, wherein the compound has the formula:
Figure imgf000121_0001
wherein:
R7 represents from O to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and
(ii) Ci-C6alkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Cj-Cόalkoxycarbonyl, C2- Cόalkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
/S)
^~^^ is a 5- or 6-membered heterocycle that is optionally substituted with Ci-C4alkyl.
4. The compound or salt or hydrate thereof according to claim 3, wherein the compound has the formula:
Figure imgf000121_0002
5. A compound or salt or hydrate thereof according to claim 3 or claim 4, wherein R7 represents two methyl substituents or a spiro C3-C6cycloalkyl.
6. The compound or salt or hydrate thereof according to claim 2 wherein the compound is
Figure imgf000121_0003
wherein: o is 0, 1 or 2; R7 represents from O to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and
(ii) Ci-C6alkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Ci-Cόalkoxycarbonyl, C2- C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
M is NH, O, S, SO, SO2 or CH2.
7. The compound or salt or hydrate thereof according to claim 6, wherein R7 represents from 0 to 2 substituents independently chosen from hydroxy, amino, oxo, d-C6alkyl, Ci- Qhydroxyalkyl, d-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, (Ci-C6alkanoyl)amino, Ci- C6alkoxycarbonyl, C2-C6alkyl ether, phenyl and 4- to 7-membered heterocycles.
8. The compound or salt or hydrate thereof according to any one of claims 2-7, wherein:
Figure imgf000122_0001
9. The compound or salt or hydrate thereof according to claim 8, wherein the compound has the formula:
Figure imgf000122_0002
wherein each R9 is independently hydrogen or Ci-C4alkyl, or both R9 are taken together to form a spiro C3-C8cycloalkyl.
10. The compound or salt or hydrate thereof according to any one of claims 1-9, wherein R3 represents from 0 to 2 substituents independently chosen from halogen, Ci-C4alkyl, Ci- C4hydroxyalkyl, Ci-C4alkoxy, C2-C4alkyl ether and mono- or di-(Ci-C4alkyl)amino.
11. The compound or salt or hydrate thereof according to claim 8, wherein:
Figure imgf000123_0001
is , wherein represents a 5- or 6-membered carbocycle or heterocycle, each of which is substituted with from 0 to 4 substituents independently chosen from halogen, hydroxy, amino, Ci-C6alkyl, (C3-C8carbocycle)Co-C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, and (4- to 7-membered heterocycle)C0-C4alkyl.
12. The compound or salt or hydrate thereof according to any one of claims 2-11, wherein n is O.
13. The compound or salt or hydrate thereof according to claim 12, wherein the compound has the formula:
or
Figure imgf000123_0002
wherein:
Yi, Y2, Y3, Y4 and Y5 are independently N, C, or CH;
R53 is hydrogen, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl or (C3-C8cycloalkyl)C0-C4alkyl; and
R8 represents from O to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
14. The compound or salt or hydrate thereof according to any one of claims 2-11, wherein n is l.
15. The compound or salt or hydrate thereof according to claim 14, wherein the compound has the formula:
Figure imgf000124_0001
wherein:
Yi, Y2, Y3 and Y4 are independently N, C, or CH; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
16. The compound or salt or hydrate thereof according to any one of claims 2-11, wherein n is 2.
17. The compound or salt or hydrate thereof according to claims 16, wherein the compound has the formula:
Figure imgf000124_0002
wherein:
Yi, Y2, Y3 and Y4 are independently N, C, or CH; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
18. The compound or salt or hydrate thereof according to any one of claims 2-17, wherein R5 and R6 each represent 0 substituents.
19. The compound or salt or hydrate thereof according to claim 1 wherein the compound is:
Figure imgf000124_0003
wherein:
Figure imgf000125_0001
and V are independently C, CH, N, NH, S, and O, such that at least one of T, U and V is N; exactly one of T, U and V is CH; and
Figure imgf000125_0002
Figure imgf000125_0003
wherein
X is N;
Z is C, CH, CH2, N or NH; ^^ represents a single or double bond; n is 0, 1 or 2; R4 represents: (i) oxo
(ii) a phenyl or 5- or 6-membered heteroaryl substituent; or (iii) a fused phenyl or 5- or 6-membered heterocyclic ring; each of which (ii) and (iii) is substituted with from 0 to 4 substituents independently chosen from:
(a) halogen, hydroxy, cyano, amino and oxo; and
(b) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, Ci- C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8cycloalkyl)C0-C4alkyl, mono- or di- (Ci- C6alkyl)aminoCo-C4alkyl, and (4- to 10-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino, aminosulfonyl, Ci-C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-C6alkanoyl, Ci- C6alkylsulfonyl, Ci-C6alkylsulfinyl, Ci-C6alkoxycarbonyl, mono- or di-(Ci- C6alkyl)aminocarbonyl, and mono- or di-(C]-C6alkyl)aminosulfonyl; and
R5 represents from 0 to 4 substituents independently chosen from Ci-Cβalkyl, C2- C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl.
20. The compound or salt or hydrate thereof according to claim 19, wherein the compound has the formula:
Figure imgf000126_0001
wherein:
R7 represents from O to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and
(ii) Ci-C6alkyl, C]-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Ci-Cόalkoxycarbonyl, C2- Cόalkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, C]-C4alkoxy and mono- or di-(C]-C4alkyl)amino; and
Figure imgf000126_0002
is a 5- or 6-membered heterocycle that is optionally substituted with Ci-C4alkyl.
21. The compound or salt or hydrate thereof according to claim 20, wherein the compound has the formula:
Figure imgf000126_0003
22. The compound or salt or hydrate thereof according to claim 20 or claim 21, wherein R7 represents two methyl substituents or a spiro C3-C6cycloalkyl.
23. The compound or salt or hydrate thereof according to claim 19, wherein the compound has the formula:
Figure imgf000126_0004
wherein: o is 0, 1 or 2;
R7 represents from 0 to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and (ii) Ci-C6alkyl, d-Cόalkoxy, mono- or di-(Ci-C6alkyl)amino, Ci-C6alkoxycarbonyl, C2- C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
M is NH, O, S, SO, SO2 or CH2.
24. The compound or salt or hydrate thereof according to claim 23, wherein R7 represents from 0 to 2 substituents independently chosen from hydroxy, amino, oxo, C]-C6alkyl, C]- C6hydroxyalkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, (Ci-C6alkanoyl)amino, Ci- C6alkoxycarbonyl, C2-Cόalkyl ether, phenyl and 4- to 7-membered heterocycles.
25. The compound or salt or hydrate thereof according to any one of claims 19-24, wherein:
Figure imgf000127_0001
; wherein G is NH,
O or S.
26. The compound or salt or hydrate thereof according to claim 25, wherein:
Figure imgf000127_0002
wherein R3a is hydrogen, Ci-C4alkyl, Ci-C4hydroxyalkyl or C2-C4alkyl ether.
27. The compound or salt or hydrate thereof according to claim 26, wherein the compound satisfies the formula:
Figure imgf000127_0003
28. The compound or salt or hydrate thereof according to any one of claims 19-25, wherein R3 represents O substituents or one methyl substituent.
29. The compound or salt or hydrate thereof according to any one of claims 19-28, wherein n is O.
30. The compound or salt or hydrate thereof according to claim 29, wherein the compound has the formula:
Figure imgf000128_0001
wherein:
Yi, Y2, Y3, Y4 and Y5 are independently N, C, or CH; R5a is hydrogen, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl or (C3-Cgcycloalkyl)Co-C4alkyl; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ct- Cβalkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
31. The compound or salt or hydrate thereof according to any one of claims 19-28, wherein n is l.
32. The compound or salt or hydrate thereof according to claim 31, wherein the compound has the formula:
Figure imgf000128_0002
wherein:
Yi, Y2, Y3 and Y4 are independently N, C, or CH; and R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Q-Cβalkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)Co-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
33. The compound or salt or hydrate thereof according to any one of claims 19-28, wherein n is 2.
34. The compound or salt or hydrate thereof according to claim 33, wherein the compound has the formula:
Figure imgf000129_0001
wherein:
Yi, Y2, Y3 and Y4 are independently N, C, or CH; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)Co-C4alkyl, Ci- Qalkoxy, and mono- or di-(Ci-C6alkyl)aminoCo-C4alkyl.
35. The compound or salt or hydrate thereof according to any one of claims 19-34, wherein R5 and R6 each represent 0 substituents.
36. The compound or salt or hydrate thereof according to claim 1 wherein the compound is:
Figure imgf000129_0002
or a pharmaceutically acceptable salt or hydrate thereof, wherein:
Ar is B-D R3 wherein A, B, D and E are each CH, C, or N, such that exactly one or two of A, B, D and E are N;
Figure imgf000130_0001
wherein
X is C or CH; Z is C, CH, CH2, N or NH; each ^^ represents a single or double bond; n is 0, 1 or 2; R4 represents: (i) oxo;
(ii) a phenyl or 5- or 6-membered heteroaryl substituent; or (iii) a fused phenyl or 5- or 6-membered heterocyclic ring; each of which (ii) and (iii) is substituted with from 0 to 4 substituents independently chosen from:
(a) halogen, hydroxy, cyano, amino and oxo; and
(b) C,-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Q-Qhaloalkyl, Ci-C6hydroxyalkyl, Cr C6alkoxy, C]-C6alkoxycarbonyl, (C3-C8cycloalkyl)C0-C4alkyl, mono- or di- (Ci- Cόalkyl)aminoC0-C4alkyl, and (4- to 10-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino, aminocarbonyl, aminosulfonyl, C]-C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-Cόalkanoyl, C]-C6alkylsulfonyl, C]-C6alkylsulfinyl, Ci-C6alkoxycarbonyl, mono- or di-(Ci- C6alkyl)aminocarbonyl, and mono- or di-(Ci-C6alkyl)aminosulfonyl; and
R5 represents from 0 to 4 substituents independently chosen from Ci-C6alkyl, C2- C6alkenyl, C2-C6alkynyl, and (C3-C8cycloalkyl)C0-C4alkyl.
37. The compound or salt or hydrate thereof according to claim 36, wherein the compound has the formula:
Figure imgf000130_0002
R7 represents from 0 to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and (ii) Ci-C6alkyl, Ci-C6alkoxy, mono- or di-(C]-C6alkyl)amino, Ci-C6alkoxycarbonyl, C2- C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
Figure imgf000131_0001
is a 5- or 6-membered heterocycle that is optionally substituted with Ci-C4alkyl.
38. The compound or salt or hydrate thereof according to claim 37, wherein the compound has the formula:
Figure imgf000131_0002
39. The compound or salt or hydrate thereof according to claim 37 or claim 38, wherein R7 represents two methyl substituents or a spiro C3-C6cycloalkyl.
40. The compound or salt or hydrate thereof according to claim 37, wherein the compound has the formula:
Figure imgf000131_0003
wherein: o is 0, 1 or 2;
R7 represents from 0 to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and
(ii) Ci-C6alkyl, Ci-C6alkoxy, mono- or di-(C]-C6alkyl)amino, Ci-C6alkoxycarbonyl, C2- C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
M is NH, O, S, SO, SO2 or CH2.
41. The compound or salt or hydrate thereof according to claim 40, wherein R7 represents from 0 to 2 substituents independently chosen from hydroxy, amino, oxo, Ci-C6alkyl, Ci- C6hydroxyalkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, (Ci-C6alkanoyl)amino, Cj- C6alkoxycarbonyl, C2-C6alkyl ether, phenyl and 4- to 7-membered heterocycles.
42. The compound or salt or hydrate thereof according to any one of claims 36-41, wherein:
Figure imgf000132_0001
43. The compound or salt or hydrate thereof according to claim 42, wherein the compound has the formula:
Figure imgf000132_0002
wherein each R9 is independently hydrogen or Ci-C4alkyl, or both R9 are taken together to form a spiro C3-C8cycloalkyl
44. The compound or salt or hydrate thereof according to any one of claims 36-43, wherein R3 represents from 0 to 2 substituents independently chosen from halogen, Ci-C4alkyl, Ci- C4hydroxyalkyl, Ci-C4alkoxy, C2-C4alkyl ether and mono- or di-(Ci-C4alkyl)amino.
45. The compound or salt or hydrate thereof according to claim 42, wherein:
Figure imgf000132_0003
is , wherein represents a 5- or 6-membered carbocycle or heterocycle, each of which is substituted with from 0 to 4 substituents independently chosen from halogen, hydroxy, amino, Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, and (4- to 7-membered heterocycle)Co-C4alkyl.
46. The compound or salt or hydrate thereof according to any one of claims 36-45, wherein n is O.
47. The compound or salt or hydrate thereof according to claim 46, wherein the compound has the formula:
Figure imgf000133_0001
wherein:
Yi, Y2, Y3, Y4 and Y5 are independently N, C, or CH;
R.5a is hydrogen, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl or (C3-C8cycloalkyl)C0-C4alkyl; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Cj-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, C1- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
48. The compound or salt or hydrate thereof according to any one of claims 36-45, wherein n is l.
49. The compound or salt or hydrate thereof according to claim 48, wherein the compound has the formula:
Figure imgf000133_0002
wherein:
Yi, Y2, Y3 and Y4 are independently N, C, or CH; and R8 represents from O to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)Co-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
50. The compound or salt or hydrate thereof according to any one of claims 36-45, wherein n is 2.
51. The compound or salt or hydrate thereof according to claim 50, wherein the compound has the formula:
Figure imgf000134_0001
wherein:
Y1, Y2, Y3 and Y4 are independently N, C, or CH; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, C]-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)Co-C4alkyl, Cj- Qalkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
52. The compound or salt or hydrate thereof according to any one of claims 36-51, wherein R5 and R6 each represent 0 substituents.
53. The compound or salt or hydrate thereof according to claim 1 wherein the compound is:
Figure imgf000134_0002
or a pharmaceutically acceptable salt or hydrate thereof, wherein:
Ar is
Figure imgf000134_0003
wherein T, U and V are independently C, CH, N, NH, S and O, such
that at least one of T, U and V is N; exactly one of T, U and V is CH; and V R 3 is aromatic;
Figure imgf000135_0001
wherein
X is C or CH; Z is C, CH, CH2, N or NH; ^^ represents a single or double bond; n is 0, 1 or 2; R4 represents: (i) oxo
(ii) a phenyl or 5- or 6-membered heteroaryl substituent; or (iii) a fused phenyl or 5- or 6-membered heterocyclic ring; each of which (ii) and (iii) is substituted with from 0 to 4 substituents independently chosen from:
(a) halogen, hydroxy, cyano, amino and oxo; and
(b) Ci-Cealkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, d-C6hydroxyalkyl, Ci- C6alkoxy, C]-C6alkoxycarbonyl, (C3-Cgcycloalkyl)C0-C4alkyl, mono- or di- (C]- C6alkyl)aminoC0-C4alkyl, and (4- to 10-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino, aminosulfonyl, Ci-C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-C6alkanoyl, Ci- Cόalkylsulfonyl, Ci-C6alkylsulfinyl, Ci-C6alkoxycarbonyl, mono- or (Ii-(C1- C6alkyl)aminocarbonyl, and mono- or di-(Ci-C6alkyl)aminosulfonyl; and
R5 represents from 0 to 4 substituents independently chosen from Ci-C6alkyl, C2- C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl.
54. The compound or salt or hydrate thereof according to claim 53, wherein the compound has the formula:
Figure imgf000135_0002
wherein:
R7 represents from 0 to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and (ii) Ci-C6alkyl, C]-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Ci-C6alkoxycarbonyl, C2- C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
Figure imgf000136_0001
— is a 5- or 6-membered heterocycle that is optionally substituted with Ci-C4alkyl.
55. The compound or salt or hydrate thereof according to claim 54, wherein the compound has the formula:
Figure imgf000136_0002
56. The compound or salt or hydrate thereof according to claim 54 or claim 55, wherein R7 represents two methyl substituents or a spiro C3-C6cycloalkyl.
57. A compound or salt or hydrate thereof according to claim 53, wherein the compound has the formula:
Figure imgf000136_0003
wherein:
X is CH; o is 0, 1 or 2;
R7 represents from 0 to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and
(ii) Ci-C6alkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Ci-C6alkoxycarbonyl, C2- C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
M is NH, O, S, SO, SO2 or CH2.
58. The compound or salt or hydrate thereof according to claim 57, wherein R7 represents from 0 to 2 substituents independently chosen from hydroxy, amino, oxo, Ci-C6alkyl, Ci- C6hydroxyalkyl, C]-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, (Ci-C6alkanoyl)amino, Ci- C6alkoxycarbonyl, C2-C6alkyl ether, phenyl and 4- to 7-membered heterocycles.
59. The compound or salt or hydrate thereof according to any one of claims 53-58, wherein:
Figure imgf000137_0001
O or S.
60. The compound or salt or hydrate thereof according to claim 59, wherein:
Figure imgf000137_0002
wherein R3a is hydrogen, Ci-C4alkyl, Ci-C4hydroxyalkyl or C2-C4alkyl ether.
61. The compound or salt or hydrate thereof according to claim 60, wherein the compound satisfies the formula:
Figure imgf000137_0003
62. The compound or salt or hydrate thereof according to any one of claims 53-61, wherein R3 represents 0 substituents or one methyl substituent.
63. The compound or salt or hydrate thereof according to any one of claims 53-62, wherein n is O.
64. The compound or salt or hydrate thereof according to claim 63, wherein the compound has the formula:
Figure imgf000138_0001
wherein:
Yi, Y2, Y3, Y4 and Y5 are independently N, C, or CH;
R.5a is hydrogen, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl or (C3-C8cycloalkyl)C0-C4alkyl; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoCo-C4alkyl.
65. The compound or salt or hydrate thereof according to any one of claims 53-62, wherein n is l.
66. The compound or salt or hydrate thereof according to claim 65, wherein the compound has the formula:
Figure imgf000138_0002
wherein:
Y1, Y2, Y3 and Y4 are independently N, C, or CH; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, C1- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
67. The compound or salt or hydrate thereof according to any one of claims 53-62, wherein n is 2.
68. The compound or salt or hydrate thereof according to claim 67, wherein the compound has the formula:
Figure imgf000139_0001
wherein:
Yi, Y2, Y3 and Y4 are independently N, C, or CH; and
R8 represents from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)Co-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
69. The compound or salt or hydrate thereof according to any one of claims 53-68, wherein R5 and R6 each represent 0 substituents.
70. A compound of the formula:
Figure imgf000139_0002
or a pharmaceutically acceptable salt or hydrate thereof, wherein:
P is a 5- or 6-membered heterocycle that:
(a) comprises at least one ring nitrogen atom;
(b) is substituted with oxo;
(c) is optionally substituted with Ci-C4alkyl; and
(d) is:
(i) substituted with a phenyl or 5- or 6-membered heteroaryl substituent; or
(ii) fused to a phenyl or 5- or 6-membered heterocyclic ring; each of which (i) and (ii) is substituted with from 0 to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3- C8cycloalkyl)Co-C4alkyl, C]-C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl;
Ar is a 5- or 6-membered heteroaryl that is substituted with from 0 to 2 substituents independently chosen from R3; Ri is Ci-C8aminoalkyl, Ci-Cghydroxyalkyl, C2-C8alkyl ether, phenylC0-C6alkyl, or (4- to 12-membered heterocycle)C0-C6alkyl, each of which is substituted with from 0 to 6 substituents independently chosen from:
(i) halogen, hydroxy, cyano, amino, aminocarbonyl, aminosulfonyl and oxo; and
(ii) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, Ci- C6alkoxy, Ci-C6alkoxycarbonyl, (C3-C8carbocycle)C0-C4alkyl, mono- or di- (Ci- C6alkyl)aminoC0-C4alkyl, and (4- to 12-membered heterocycle)Co-C4alkyl, each of which is substituted with from 0 to 4 substituents independently chosen from hydroxy, oxo, halogen, amino and Ci-C4alkyl; and
R2 is hydrogen, Ci-C6alkyl or Ci-C6haloalkyl; or Ri and R2 are taken together to form a 4- to 10-membered heterocycle that is substituted with from 0 to 6 substituents independently chosen from:
(i) halogen, hydroxy, cyano, amino, aminocarbonyl, aminosulfonyl and oxo; and
(ii) Ci-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, Ci-C6haloalkyl, Ci-C6hydroxyalkyl, C2- C6alkyl ether, C]-C6alkoxy, Ci-C6alkoxycarbonyl, (C3-Cgcycloalkyl)Co-C4alkyl, mono- or di- (Ci-C6alkyl)aminoC0-C4alkyl, phenylC0-C4alkyl, (4- to 12-membered heterocycle)C0-C4alkyl, and spiro C3-Cscycloalkyl; each of which is substituted with from 0 to 6 substituents independently chosen from hydroxy, oxo, halogen, amino, aminocarbonyl, aminosulfonyl, Ci- C6alkyl, Ci-C6alkoxy, Ci-C6alkylthio, Ci-C6alkanoyl, Ci-C6alkylsulfonyl, Ci-C6alkylsulfmyl, Ci-C6alkoxycarbonyl, mono- or di-(Ci-Cόalkyl)aminocarbonyl, and mono- or di-(Ci- Cόalkyl) aminosulfonyl ;
Each R3 is independently chosen from:
(i) halogen, hydroxy, cyano and amino; and
(ii) Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-C6haloalkyl, d-C6alkoxy, Cj-C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, or (4- to 7-membered heterocycle)C0-C4alkyl; each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, Ci-C4alkyl, (C3-C8carbocycle)Co-C4alkyl and (4- to 10-membered heterocycle)C0-C4alkyl; or two substituents represented by R3 are taken together to form a fused 5- to 7- membered carbocycle or heterocycle that is substituted with from 0 to 3 substituents independently chosen from halogen, hydroxy, amino, Ci-C6alkyl, (C3-C8carbocycle)C0-C4alkyl, Ci-C6haloalkyl, Ci-C6alkoxy, Ci-C6alkyl ether, mono- or di-(Ci-C6alkyl)aminoC0-C2alkyl, and (4- to 7-membered heterocycle)C0-C4alkyl; and R6 represents from O to 4 substituents independently chosen from halogen, hydroxy, amino, cyano, Ci-C6alkyl, d-C6haloalkyl, Ci-C6alkoxy and Ci-C6alkyl ether.
71. The compound or salt or hydrate thereof according to claim 70, wherein the compound has the formula:
Figure imgf000141_0001
wherein:
A, B, D and E are each CH, C, or N, such that exactly one or two of A, B, D and E are N; T, U and V are independently C, CH, N, NH, S and O, such that at least one of T, U and
V is N; exactly one of T, U and V is CH; and
Figure imgf000141_0002
is aromatic.
72. The compound or salt or hydrate thereof according to claim 70 or claim 71, the compound has the formula:
Figure imgf000141_0003
wherein:
R7 represents from 0 to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and
(ii) Ci-C6alkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Ci-C6alkoxycarbonyl, C2- Qalkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
/S)
— is a 5- or 6-membered heterocycle that is optionally substituted with Ci-C4alkyl.
73. The compound or salt or hydrate thereof according to claim 72, wherein the compound has the formula:
Figure imgf000142_0001
74. The compound or salt or hydrate thereof according to claim 72 or claim 73, wherein R7 represents two methyl substituents or a spiro C3-C6cycloalkyl.
75. The compound or salt or hydrate thereof according to claim 70 or claim 71, wherein the compound has the formula:
Figure imgf000142_0002
wherein: o is 0, 1 or 2;
R7 represents from 0 to 4 substituents independently chosen from:
(i) halogen, hydroxy, amino and oxo; and
(ii) C]-C6alkyl, Ci-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, Ci-Cόalkoxycarbonyl, C2- C6alkyl ether, phenyl, 4- to 7-membered heterocycle and spiro C3-C7cycloalkyl, each of which is substituted with from 0 to 2 substituents independently chosen from halogen, hydroxy, amino, oxo, Ci-C4alkyl, Ci-C4alkoxy and mono- or di-(Ci-C4alkyl)amino; and
M is NH, O, S, SO, SO2 or CH2.
76. The compound or salt or hydrate thereof according to claim 75, wherein R7 represents from 0 to 2 substituents independently chosen from hydroxy, amino, oxo, Ci-C6alkyl, Ci- C6hydroxyalkyl, Cj-C6alkoxy, mono- or di-(Ci-C6alkyl)amino, (Ci-C6alkanoyl)amino, Q- C6alkoxycarbonyl, C2-C6alkyl ether, phenyl and 4- to 7-membered heterocycles.
77. The compound or salt or hydrate thereof according to any one of claims 70-76, wherein:
Figure imgf000143_0001
wherein:
Y1, Y2, Y3, Y4 and Y5 are independently N, C, or CH; and
R8 represents from O to 4 substituents independently chosen from halogen, hydroxy, cyano, amino, C]-C6alkyl, C2-C6alkenyl, C2-C6alkynyl, (C3-C8cycloalkyl)C0-C4alkyl, Ci- C6alkoxy, and mono- or di-(Ci-C6alkyl)aminoC0-C4alkyl.
78. The compound or salt or hydrate thereof according to any one of claims 70-77, wherein R3 represents from 0 to 2 substituents independently chosen from halogen, Ci-C4alkyl, Ci- C4hydroxyalkyl, Ci-C4alkoxy, C2-C4alkyl ether and mono- or di-(Ci-C4alkyl)amino.
79. The compound or salt or hydrate thereof according to any one of claims 70-78, wherein R6 represents 0 substituents.
80. A pharmaceutical composition, comprising at least one compound or salt or hydrate thereof according to any one of claims 1-79, in combination with a physiologically acceptable carrier or excipient.
81. The pharmaceutical composition according to claim 80, wherein the composition is formulated as an injectible fluid, an aerosol, a cream, a gel, a pill, a capsule, a syrup or a transdermal patch.
82. A method for inhibiting ligand-induced CGRP receptor activity in vitro, the method comprising contacting a CGRP receptor with at least one compound or salt or hydrate thereof according to any one of claims 1-79, under conditions and in an amount sufficient to detectably inhibit ligand-induced CGRP receptor activity.
83. A method for inhibiting induction of ligand-induced CGRP receptor activity in a patient, comprising contacting cells expressing a CGRP receptor with at least one compound or salt or hydrate thereof according to any one of claims 1-79, in an amount sufficient to detectably inhibit ligand-induced CGRP receptor activity in cells expressing a cloned CGRP receptor in vitro, and thereby inhibiting ligand-induced CGRP activity in the patient.
84. The method according to claim 83, wherein the patient is a human.
85. A method for treating a condition responsive to CGRP receptor modulation in a patient, comprising administering to the patient a therapeutically effective amount of at least one compound or salt or hydrate thereof according to any one of claims 1-79, and thereby alleviating the condition in the patient.
86. The method according to claim 85, wherein the condition is headache, inflammation, neurogenic vasodilation, thermal injury, shock or hot flashes.
87. A method for treating migraine in a patient, comprising administering to a patient suffering from migraine a therapeutically effective amount of at least one compound or salt or hydrate thereof according to any one of claims 1-79, and thereby alleviating migraine in the patient.
88. A method for treating headache in a patient, comprising administering to a patient suffering from headache a therapeutically effective amount of at least one compound or salt or hydrate thereof according to any one of claims 1-79, and thereby alleviating headache in the patient.
89. The method according to claim 88, wherein the patient is suffering from cluster headache or tension headache.
90. A method according to claim 88 or claim 89, wherein the patient is a human.
91. A compound or salt or hydrate thereof according to any one of claims 1-79, wherein the compound is radiolabeled.
92. A method for determining the presence or absence of CGRP receptor in a sample, comprising the steps of:
(a) contacting a sample with a compound or salt or hydrate thereof according to any one of claims 1-79, under conditions that permit binding of the compound to CGRP receptor; and
(b) detecting a signal indicative of a level of the compound bound to CGRP receptor, and therefrom determining the presence or absence of CGRP receptor in the sample.
93. The method according to claim 92, wherein the compound is radiolabeled, and wherein the step of detection comprises the steps of:
(i) separating unbound compound from bound compound; and
(ii) detecting the presence or absence of bound radiolabel in the sample.
94. A packaged pharmaceutical preparation, comprising:
(a) a pharmaceutical composition according to claim 80 in a container; and
(b) instructions for using the composition to treat a condition responsive to CGRP receptor modulation.
95. The packaged pharmaceutical preparation according to claim 94, wherein the condition responsive to CGRP receptor modulation is migraine or headache.
96. A use of a compound or salt or hydrate thereof according to any one of claims 1-79 for the manufacture of a medicament for the treatment of a condition responsive to CGRP receptor modulation.
97. The use according to claim 96, wherein the condition is migraine or headache.
98. A method for detecting CGRP receptor antagonist activity of a test compound, comprising:
(i) contacting test cells that stably expresses CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2 with:
(a) a test compound;
(b) an indicator of the level of intracellular calcium; and
(c) CGRP receptor agonist; (ii) detecting a response indicative of intracellular calcium level in the contacted test cells;
(iii) contacting matched control cells that stably expresses CGRP receptor and rat olfactory cyclic nucleotide gated channel subunits 1 and 2 with:
(a) an indicator of the level of intracellular calcium; and
(b) CGRP receptor agonist;
(iv) detecting a response indicative of intracellular calcium level in the contacted control cells; and
(iii) comparing the responses detected in (ii) and (iv); and therefrom detecting CGRP receptor antagonist activity of the test compound.
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