The present invention relates to compounds that affect cholinergic receptors, especially muscarinic receptors. The present invention provides compounds that are agonists of cholinergic receptors including muscarinic receptors, especially the m1 and m4 subtype of muscarinic receptors. The invention also provides methods of using the provided compounds for modulating conditions associated with cholinergic receptors, especially for treating or alleviating disease conditions associated with muscarinic receptors, e.g., m1 or m4 subtypes of receptors.
Muscarinic cholinergic receptors mediate the actions of the neurotransmitter acetylcholine in the central and peripheral nervous systems, gastrointestinal system, heart, endocrine glands, lungs, and other tissues. Muscarinic receptors play a central role in the central nervous system for higher cognitive functions, as well as in the peripheral parasympathetic nervous system. Five distinct muscarinic receptor subtypes have been identified, m1-m5. The m1 subtype is the predominant subtype found in the cerebral cortex and is believed to be involved in the control of cognitive functions; m2 is the predominant subtype found in heart and is believed to be involved in the control of heart rate; m3 is believed to be involved in gastrointestinal and urinary tract stimulation as well as sweating and salivation; m4 is present in brain and may be involved in locomotion; and m5, present in brain, may be involved in certain functions of the central nervous system associated with the dopaminergic system.
Conditions associated with cognitive impairment, such as Alzheimer""s disease, are accompanied by loss of acetylcholine in the brain. This is believed to be the result of degeneration of cholinergic neurons in the basal forebrain, which innervate areas of the association cortex, and hippocampus, which is involved in higher processes.
Efforts to increase acetylcholine levels have focused on increasing levels of choline, the precursor for acetylcholine synthesis, and on blocking acetylcholine esterase (AChE), the enzyme that metabolizes acetylcholine. Administration of choline or phosphatidylcholine has not been very successful. AChE inhibitors have shown some therapeutic efficacy, but may cause cholinergic side effects due to peripheral acetylcholine stimulation, including abdominal cramps, nausea, vomiting, diarrhea, anorexia, weight loss, myopathy and depression. Gastrointestinal side effects have been observed in about a third of the patients treated. In addition, some AChE inhibitors, such as tacrine, have also been found to cause significant hepatotoxicity, with elevated liver transaminases observed in about 30% of patients. The adverse effects of AChE inhibitors have limited their clinical utility.
Known m1 muscarinic agonists such as arecoline have also been found to be weak agonists of m2 as well as m3 subtype and are not very effective in treating cognitive impairment, most likely because of dose-limiting side effects.
There is a need for compounds that increase acetylcholine signaling or effect in the brain. Specifically there is a need for muscarinic agonists that are active at various muscarinic receptor subtypes in the central and peripheral nervous system. Furthermore, there is a need for more highly selective muscarinic agonists, such as m1- or m4-selective agents, both as pharmacological tools and as therapeutic agents.
The present invention provides compounds that affect cholinergic, especially muscarinic, receptors that have agonist activity at the m1 or m4 subtype of muscarinic receptors, or both. The compounds of the invention are of the general formula (I): 
wherein:
Z1 is CR1 or N, Z2 is CR2 or N, Z3 is CR3 or N, and Z4 is CR4 or N, where no more than two of Z1, Z2, Z3 and Z4 are N;
W1 is O, S, or NR5, one of W2 and W3 is N or CR6, and the other of W2 and W3 is CG; W1 is NG, W2 is CR5 or N, and W3 is CR6 or N; or W1 and W3 are N, and W2 is NG;
G is of formula (II): 
Y is O, S, CHOH, xe2x80x94NHC(O)xe2x80x94, xe2x80x94C(O)NHxe2x80x94, xe2x80x94C(O)xe2x80x94, xe2x80x94OC(O)xe2x80x94, xe2x80x94(O)COxe2x80x94, xe2x80x94NR7xe2x80x94, xe2x80x94CHxe2x95x90Nxe2x80x94, or absent;
p is 1, 2, 3, 4 or 5;
Z is CR8R9 or absent;
each t is 1, 2, or 3;
each R1, R2, R3, and R4, independently, is H, amino, hydroxyl, halo, or straight- or branched-chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 heteroalkyl, C1-6 haloalkyl, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OR11, xe2x80x94COR11, xe2x80x94NO2, xe2x80x94SR11, xe2x80x94NHC(O)R11, xe2x80x94C(O)NR12R13, xe2x80x94NR12R13, xe2x80x94NR11C(O)NR12R13, xe2x80x94SO2NR12R13, xe2x80x94OC(O)R11, xe2x80x94O(CH2)qNR12R13, or xe2x80x94(CH2)qNR12R13, where q is an integer from 2 to 6, or R1 and R2 together form xe2x80x94NHxe2x80x94Nxe2x95x90Nxe2x80x94 or R3 and R4 together form xe2x80x94NHxe2x80x94Nxe2x95x90Nxe2x80x94;
each R5, R6, and R7, independently, is H, C1-6 alkyl; formyl; C3-6 cycloalkyl; C5-6 aryl, optionally substituted with halo or C1-6 alkyl; or C5-6 heteroaryl, optionally substituted with halo or C1-6 alkyl;
each R8 and R9, independently, is H or straight- or branched-chain C1-8 alkyl;
R10 is straight- or branched-chain C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C1-8 alkylidene, C1-8 alkoxy, C1-8 heteroalkyl, C1-8 aminoalkyl, C1-8 haloalkyl, C1-8 alkoxycarbonyl, C1-8 hydroxyalkoxy, C1-8 hydroxyalkyl, xe2x80x94SH, C1-8 alkylthio, xe2x80x94Oxe2x80x94CH2xe2x80x94C5-6 aryl, xe2x80x94C(O)xe2x80x94C5-6 aryl substituted with C1-3 alkyl or halo, C5-6 aryl, C5-6 cycloalkyl, C5-6 heteroaryl, C5-6 heterocycloalkyl, xe2x80x94NR12R13, xe2x80x94C(O)NR12R13, xe2x80x94NR11C(O)NR12R13, xe2x80x94CR11R12R13,xe2x80x94OC(O)R11, xe2x80x94(O)(CH2)SNR12R13 or xe2x80x94(CH2)SNR12R13, s being an integer from 2 to 8;
R10xe2x80x2 is H, straight- or branched-chain C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C1-8 alkylidene, C1-8 alkoxy, C1-8 heteroalkyl, C1-8 aminoalkyl, C1-8 haloalkyl, C1-8 alkoxycarbonyl, C1-8 hydroxyalkoxy, C1-8 hydroxyalkyl, or C1-8 alkylthio;
each R11, independently, is H, straight- or branched-chain C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C2-8 heteroalkyl, C2-8 aminoalkyl, C2-8 haloalkyl, C1-8 alkoxycarbonyl, C2-8 hydroxyalkyl, xe2x80x94C(O)xe2x80x94C5-6 aryl substituted with C1-3 alkyl or halo, C5-6 aryl, C5-6 heteroaryl, C5-6 cycloalkyl, C5-6 heterocycloalkyl, xe2x80x94C(O)NR12R13, xe2x80x94CR5R12R13, xe2x80x94(CH2)tNR12R13, t is an integer from 2 to 8; and
each R12 and R13, independently, is H, C1-6 alkyl; C3-6 cycloalkyl; C5-6 aryl, optionally substituted with halo or C1-6 alkyl; or C5-6 heteroaryl, optionally substituted with halo or C1-6 alkyl; or R12 and R13 together form a cyclic structure;
or a pharmaceutically acceptable salt, ester or prodrug thereof.
The present invention further provides pharmaceutical compositions including an effective amount of a compound of formula (I) or pharmaceutically acceptable salts, esters, or prodrugs thereof.
Also provided are methods of increasing an activity of a cholinergic receptor comprising contacting the cholinergic receptor or a system containing the cholinergic receptor with an effective amount of a compound of formula (I), as well as kits for performing the same. Preferably, the receptor is a muscarinic receptor of the m1 or m4 subtype. The receptor may be located in the central nervous system, peripheral nervous system, gastrointestinal system, heart, endocrine glands, or lungs; and the receptor may be a truncated, mutated, or modified cholinergic receptor.
Furthermore, the present invention relates to a method of activating a cholinergic receptor comprising contacting the cholinergic receptor or a system containing the cholinergic receptor with an effective amount of at least one compound of formula (I), as well as kits for performing the method. In a preferred embodiment, the compound is selective for the m1 or m4 muscarinic receptor subtype, or both. In another preferred embodiment, the compound has little or substantially no effect on m2 or m3 activity.
Another aspect of the present invention relates to a method of treating a disease condition associated with a cholinergic receptor comprising administering to a subject in need of such treatment an effective amount of at least one of the compounds of the invention. Kits for performing the method are also provided. The disease conditions that are treated include, but are not limited to conditions of cognitive dysfunction, forgetfulness, confusion, memory loss, attention deficits, deficits in visual perception, depression, pain, sleep disorders, and psychosis. The disease conditions also include, but are not limited to diseases of Alzheimer""s disease, Parkinson""s disease, Huntington""s chorea, Friederich""s ataxia, Gilles de la Tourette""s Syndrome, Down Syndrome, Pick disease, dementia pugilistica, clinical depression, age-related cognitive decline, attention-deficit disorder, and sudden infant death syndrome.
Further provided are methods of treating the symptoms of a disease or condition associated with reduced levels of acetylcholine comprising administering an effective amount of at least one compound of the invention.
In yet another embodiment, the present invention provides a method of treating Alzheimer""s disease. The method comprises administering to a subject in need of such treatment an effective amount of at least one compound of the invention.
In still another embodiment, the present invention provides a method of treating glaucoma. The method comprises administering an effective amount of at least one compound of the invention.
Another aspect of the present invention is a method for identifying a genetic polymorphism predisposing a subject to being responsive to a compound of the invention. The method comprises administering to a subject a therapeutically effective amount of the compound; measuring the response of said subject to the compound, thereby identifying a responsive subject having an ameliorated disease condition associated with a cholinergic receptor; and identifying a genetic polymorphism in the responsive subject, wherein the genetic polymorphism predisposes a subject to being responsive to the compound.
The present invention also features a method for identifying a subject suitable for treatment with a compound of the invention, and kits for identifying the same. The method comprises detecting the presence of a polymorphism in a subject wherein the polymorphism predisposes the subject to being responsive to the compound, and wherein the presence of the polymorphism indicates that the subject is suitable for treatment with the compound.
Definitions
For the purpose of the current disclosure, the following definitions shall in their entireties be used to define technical terms and to define the scope of the composition of matter for which protection is sought in the claims.
A xe2x80x9creceptorxe2x80x9d is intended to include any molecule present inside or on the surface of a cell that may affect cellular physiology when it is inhibited or stimulated by a ligand. Typically, a receptor comprises an extracellular domain with ligand-binding properties, a transmembrane domain that anchors the receptor in the cell membrane, and a cytoplasmic domain that generates a cellular signal in response to ligand binding (xe2x80x9csignal transductionxe2x80x9d). A receptor also includes any molecule having the characteristic structure of a receptor, but with no identifiable ligand. In addition, a receptor includes a truncated, modified, mutated receptor, or any molecule comprising partial or all of the sequences of a receptor.
xe2x80x9cLigandxe2x80x9d is intended to include any substance that interacts with a receptor.
xe2x80x9cAgonistxe2x80x9d is defined as a compound that increases the activity of a receptor when it interacts with the receptor.
The xe2x80x9cm1 receptorxe2x80x9d is defined as a receptor having an activity corresponding to the activity of the m1 muscarinic receptor subtype characterized through molecular cloning and pharmacology.
xe2x80x9cSelectivexe2x80x9d or xe2x80x9cselectivityxe2x80x9d is defined as a compound""s ability to generate a desired response from a particular receptor type, subtype, class or subclass while generating less or little response from other receptor types. xe2x80x9cSelectivexe2x80x9d or xe2x80x9cselectivityxe2x80x9d of an m1 or m4 muscarinic agonist compound means a compound""s ability to increase the activity of the m1 or m4 muscarinic receptor, respectively, while causing little or no increase in the activity of other subtypes including m3 and m5 subtypes, and preferably the m2 subtype. Compounds of the presents invention may also show selectivity toward both m1 and m4 receptors, i.e. increase the activity of both the m1 and m4 muscarinic receptors, while causing little or no increase in the activity of other subtypes including the m3 and m5 subtypes, and preferably the m2 subtype.
The term xe2x80x9csubjectxe2x80x9d refers to an animal, preferably a mammal or a human, who is the object of treatment, observation or experiment.
As used herein, xe2x80x9ccoadministrationxe2x80x9d of pharmacologically active compounds refers to the delivery of two or more separate chemical entities, whether in vitro or in vivo. Coadministration means the simultaneous delivery of separate agents; the simultaneous delivery of a mixture of agents; as well as the delivery of one agent followed by delivery of a second agent or additional agents. Agents that are coadministered are typically intended to work in conjunction with each other.
The term xe2x80x9can effective amountxe2x80x9d as used herein means an amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue, system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation or palliation of the symptoms of the disease being treated.
xe2x80x9cAlkylxe2x80x9d means a straight or branched-chain alkane group with 1-6 carbon atoms in the chain, for instance methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, etc. The term xe2x80x9cheteroalkylxe2x80x9d is intended to indicate an alkane group containing 1 or 2 heteroatoms selected from O, S or N.
xe2x80x9cAlkenylxe2x80x9d means a straight or branched-chain alkene group with 2-6 carbon atoms in the chain; the term xe2x80x9calkynylxe2x80x9d is intended to indicate a straight or branched-chain alkyne group with 2-6 carbon atoms in the chain.
The terms xe2x80x9carylxe2x80x9d and xe2x80x9ccycloalkylxe2x80x9d preferably refer to mono- and bicyclic ring structures comprising 5 to 12 carbon atoms, more preferably monocyclic rings comprising 5 to 6 carbon atoms. Where such rings comprise one or more heteroatoms, selected from N, S and O, (i.e., heterocyclic, or heteroaryl rings) such rings comprise a total of 5 to 12 atoms, more preferably 5 to 6 atoms. Heterocyclic rings include, but are not limited to, furyl, pyrrolyl, pyrazolyl, thienyl, imidazolyl, indolyl, benzofuranyl, benzothiophenyl, indazolyl, benzoimidazolyl, benzothiazolyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, pyridyl, piperidinyl, piperazinyl, pyridazinyl, pyrimidinyl, pyrazinyl, morpholinyl, oxadiazolyl, thiadiazolyl, imidazolinyl, imidazolidinyl and the like. The ring may be substituted by one or more of the groups included in the definition of R2 above. It is understood that the substituents C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 alkoxy, C1-6 heteroalkyl, C1-6 aminoalkyl, C1-6 haloalkyl or C1-6 alkoxycarbonyl may, if present, be substituted by one or more of hydroxyl, C1-4 alkoxy, halogen, cyano, amino or nitro.
As used herein, the term xe2x80x9chalogenxe2x80x9d or xe2x80x9chaloxe2x80x9d includes chlorine, fluorine, which are preferred, and iodine and bromine.
The present invention provides compounds that are agonists of cholinergic receptors including muscarinic receptors. Especially, the present invention provides compounds that are selective for the m1 or m4 muscarinic receptor subtype, or both. The compounds provided by the present invention have therapeutic effect and can be used to treat disease conditions associated with cholinergic receptors, e.g. cognitive impairment in Alzheimer""s disease, glaucoma, pain, or schizophrenia.
According to one embodiment, the present invention provides compounds of formula (I) 
wherein:
Z1, is CR1 or N, Z2 is CR2 or N, Z3 is CR3 or N, and Z4 is CR4 or N, no more than two of Z1, Z2, Z3 and Z4 being N;
W1 is O, S, or NR5, one of W2 and W3 is N or CR6 and the other of W2 and W3 is CG; W1 is NG, W2 is CR5 or N, and W3 is CR6 or N; or W1 is N, W2 is NG and W3 is N;
G is of formula (II): 
Y is O, S, CHOH, xe2x80x94NHC(O)xe2x80x94, xe2x80x94C(O)NHxe2x80x94, xe2x80x94C(O)xe2x80x94, xe2x80x94OC(O)xe2x80x94, xe2x80x94(O)COxe2x80x94, xe2x80x94NR7xe2x80x94, xe2x80x94CHxe2x95x90Nxe2x80x94, or absent;
p is 1, 2, 3, 4 or 5;
Z is CR8R9 or absent;
each t is 1, 2, or 3;
each R1, R2, R3, and R4, independently, is H, amino, hydroxyl, halo, or straight- or branched-chain C1-6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1-6 heteroalkyl, C1-6 haloalkyl, xe2x80x94CN, xe2x80x94CF3, xe2x80x94OR11, xe2x80x94COR11, xe2x80x94NO2, xe2x80x94SR11, xe2x80x94NHC(O)R11, xe2x80x94C(O)NR12R13, xe2x80x94NR12R13, xe2x80x94NR11C(O)NR12R13, xe2x80x94SO2NR12R13,xe2x80x94OC(O)R11, xe2x80x94O(CH2)qNR12R13, or xe2x80x94(CH2)qNR12R13, where q is an integer from 2 to 6, or R1 and R2 together form xe2x80x94NHxe2x80x94Nxe2x95x90Nxe2x80x94 or R3 and R4 together form xe2x80x94NHxe2x80x94Nxe2x95x90Nxe2x80x94;
each R5, R6, and R7, independently, is H, C1-6 alkyl; formyl; C3-6 cycloalkyl; C5-6 aryl, optionally substituted with halo or C1-6 alkyl; or C5-6 heteroaryl, optionally substituted with halo or C1-6 alkyl;
each R8 and R9, independently, is H or straight- or branched-chain C1-8 alkyl;
R10 is straight- or branched-chain C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C1-8 alkylidene, C1-8 alkoxy, C1-8 heteroalkyl, C1-8 aminoalkyl, C1-8 haloalkyl, C1-8 alkoxycarbonyl, C1-8 hydroxyalkoxy, C1-8 hydroxyalkyl, xe2x80x94SH, C1-8 alkylthio, xe2x80x94Oxe2x80x94CH2xe2x80x94C5-6 aryl, xe2x80x94C(O)xe2x80x94C5-6 aryl substituted with C1-3 alkyl or halo, C5-6 aryl, C5-6 cycloalkyl, C5-6 heteroaryl, C5-6 heterocycloalkyl, xe2x80x94NR12R13, xe2x80x94C(O)NR12R13, xe2x80x94NR11C(O)NR12R13, xe2x80x94CR11R12R13, xe2x80x94OC(O)R11, xe2x80x94(O)(CH2)SNR12R13 or xe2x80x94(CH2)SNR12R13, s being an inter from 2 to 8;
R10xe2x80x2 is H, straight- or branched-chain C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C1-8 alkylidene, C1-8 alkoxy, C1-8 heteroalkyl, C1-8 aminoalkyl, C1-8 haloalkyl, C1-8 alkoxycarbonyl, C1-8 hydroxyalkoxy, C1-8 hydroxyalkyl, or C1-8 alkylthio;
each R11, independently, is H, straight- or branched-chain C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C2-8 heteroalkyl, C2-8 aminoalkyl, C2-8 haloalkyl, C1-8 alkoxycarbonyl, C2-8 hydroxyalkyl, xe2x80x94C(O)xe2x80x94C5-6 aryl substituted with C1-3 alkyl or halo, C5-6 aryl, C5-6 heteroaryl, C5-6 cycloalkyl, C5-6 heterocycloalkyl, xe2x80x94C(O)NR12R13, xe2x80x94CR5R12R13, xe2x80x94(CH2)tNR12R13, t is an integer from 2 to 8; and
each R12 and R13, independently, is H, C1-6 alkyl; C3-6 cycloalkyl; C5-6 aryl, optionally substituted with halo or C1-6 alkyl; or C5-6 heteroaryl, optionally substituted with halo or C1-6 alkyl; or R12 and R13 together form a cyclic structure;
or a pharmaceutically acceptable salt, ester or prodrug thereof.
According to a preferred series of embodiments, t is 2 and R10xe2x80x2 is H.
According to one preferred series of embodiments Y is xe2x80x94C(O)xe2x80x94, xe2x80x94NHC(O)xe2x80x94, S, O, xe2x80x94OC(O)xe2x80x94 or absent. In another, R10 is alkyl, and where Z1 is CR1 or N, Z2 is CR2, Z3 is CR3 or N, and Z4 is CR4. In one embodiment, p is 2. In another, R5 is H or C1-6 alkyl.
In one embodiment each R1, R2, R3, and R4, independently, is H, halo, xe2x80x94NO2, or straight- or branched-chain C1-6 alkyl, or R1 and R2 together form xe2x80x94NHxe2x80x94Nxe2x95x90Nxe2x80x94 or R3 and R4 together form xe2x80x94NHxe2x80x94Nxe2x95x90Nxe2x80x94.
Particular embodiments of the invention include:
3-[3-(4-methoxypiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-ethoxypiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-propoxypiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-butoxypiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-methoxymethylpiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-ethoxymethylpiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-propoxymethylpiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-methylpiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-ethylpiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-indole;
3-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-indole;
3-[2-(4-methoxypiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-ethoxypiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-propoxypiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-butoxypiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-methoxymethylpiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-ethoxymethylpiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-propoxymethylpiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-methylpiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-ethylpiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-n-propylpiperidine)1-yl-ethyl]-1H-indole;
3-[2-(4-n-butylpiperidine)-1-yl-ethyl]-1H-indole;
3-[2-(4-methoxypiperidine)-1-yl-ethyl]-benzo[d]isoxazole;
3-[2-(4-butoxypiperidine)-1-yl-ethyl]-benzo[d]isoxazole;
3-[3-(4-methoxypiperidine)-1-yl-propyl]-benzo[d]isoxazole;
3-[3-(4-butoxypiperidine)-1-yl-propyl]-benzo[d]isoxazole;
3-[4-(4-methoxypiperidine)-1-yl-butyl]-benzo[d]isoxazole;
3-[4-(4-butoxypiperidine)-1-yl-butyl]-benzo[d]isoxazole;
1-[3-(4-methoxypiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-ethoxypiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-propoxypiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-butoxypiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-methoxymethylpiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-ethoxymethylpiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-propoxymethylpiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-methylpiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-ethylpiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-indole;
1-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-indole;
1-[2-(4-methoxypiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-ethoxypiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-propoxypiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-butoxypiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-methoxymethylpiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-ethoxymethylpiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-propoxymethylpiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-methylpiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-ethylpiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-n-propylpiperidine)-1-yl-ethyl]-1H-indole;
1-[2-(4-n-butylpiperidine)-1-yl-ethyl]-1H-indole;
1-[3-(4-methoxypiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-ethoxypiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-propoxypiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-butoxypiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-methoxymethylpiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-ethoxymethylpiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-propoxymethylpiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-methylpiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-ethylpiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-benzotriazole;
1-[2-(4-methoxypiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-ethoxypiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-propoxypiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-butoxypiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-methoxymethylpiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-ethoxymethylpiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-propoxymethylpiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-methylpiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-ethylpiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-n-propylpiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[2-(4-n-butylpiperidine)-1-yl-ethyl]-1H-benzotriazole;
1-[4-(4-methoxypiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-ethoxypiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-propoxypiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-butoxypiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-methoxymethylpiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-ethoxymethylpiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-propoxymethylpiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-methylpiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-ethylpiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-n-propylpiperidine)-1-yl-butyl]-1H-benzotriazole;
1-[4-(4-n-butylpiperidine)-1-yl-butyl]-1H-benzotriazole;
2-[4-(4-methylpiperidine)-1-yl-butyl]-1H-benzotriazole;
2-[4-(4-ethylpiperidine)-1-yl-butyl]-1H-benzotriazole;
2-[4-(4-n-propylpiperidine)-1-yl-butyl]-1H-benzotriazole;
2-[4-(4-n-butylpiperidine)-1-yl-butyl]-1H-benzotriazole;
2-[3-(4-methylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
2-[3-(4-ethylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
2-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
2-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
2-[2-(4-methylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
2-[2-(4-ethylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
2-[2-(4-n-propylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
2-[2-(4-n-butylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
1-(1H-benzoimidazol-2-yl)-4-(4-methylpiperidine)-butanone;
1-(H-benzoimidazol-2-yl)-4-(4-ethylpiperidine)-butanone;
1-(1H-benzoimidazol-2-yl)-4-(4-n-propylpiperidine)-butanone;
1-(1H-benzoimidazol-2-yl)-4-(4-n-butylpiperidine)-butanone;
1-(1H-benzoimidazol-2-yl)-3-(4-methylpiperidine)-propanone;
1-(1H-benzoimidazol-2-yl)-3-(4-ethylpiperidine)-propanone;
1-(1H-benzoimidazol-2-yl)-3-(4-n-propylpiperidine)-propanone;
1-(1H-benzoimidazol-2-yl)-3-(4-n-butylpiperidine)-propanone;
3-[3-(4-methylpiperidine)-1-yl-propyl]-1H-indazole;
3-[3-(4-ethylpiperidine)-1-yl-propyl]-1H-indazole;
3-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-indazole;
3-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-indazole;
1-(3-benzofuran-3-yl-propyl)-4-methyl-piperidine;
1-(3-benzofuran-3-yl-propyl)-4-ethyl-piperidine;
1-(3-benzofuran-3-yl-propyl)-4-n-propyl-piperidine;
1-(3-benzofuran-3-yl-propyl)-4-n-butyl-piperidine;
3-(3-(4-methylpiperidine)-1-yl-propyl)-benzo[d]isothiazole;
3-(3-(4-ethylpiperidine)-1-yl-propyl)-benzo[d]isothiazole;
3-(3-(4-n-propylpiperidine)-1-yl-propyl)-benzo[d]isothiazole;
3-(3-(4-n-butylpiperidine)-1-yl-propyl)-benzo[d]isothiazole;
1-[3-(4-methylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
1-[3-(4-ethylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
1-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
1-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-benzoimidazole;
1-[2-(4-methylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
1-[2-(4-ethylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
1-[2-(4-n-propylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
1-[2-(4-n-butylpiperidine)-1-yl-ethyl]-1H-benzoimidazole;
1-[2-(4-methylpiperidine)-1-yl-propyl]-1H-indazole;
1-[2-(4-ethylpiperidine)-1-yl-propyl]-1H-indazole;
1-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-indazole;
1-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-indazole;
2-[4-(4-methylpiperidine)-1-yl-butyl]-1H-benzothiazole;
2-[4-(4-ethylpiperidine)-1-yl-butyl]-1H-benzothiazole;
2-[4-(4-n-propylpiperidine)-1-yl-butyl]-1H-benzothiazole;
2-[4-(4-n-butylpiperidine)-1-yl-butyl]-1H-benzothiazole;
2-[3-(4-methylpiperidine)-1-yl-propyl]-1H-benzothiazole;
2-[3-(4-ethylpiperidine)-1-yl-propyl]-1H-benzothiazole;
2-[3-(4-n-propylpiperidine)-1-yl-propyl]-1H-benzothiazole;
2-[3-(4-n-butylpiperidine)-1-yl-propyl]-1H-benzothiazole;
2-[2-(4-methylpiperidine)-1-yl-ethyl]-1H-benzothiazole;
2-[2-(4-ethylpiperidine)-1-yl-ethyl]-1H-benzothiazole;
2-[2-(4-n-propylpiperidine)-1-yl-ethyl]-1H-benzothiazole;
2-[2-(4-n-butylpiperidine)-1-yl-ethyl]-1H-benzothiazole;
2-[3-(4-methylpiperidine)-1-yl-propyl]-benzooxazole;
2-[3-(4-ethylpiperidine)-1-yl-propyl]-benzooxazole;
2-[3-(4-n-propylpiperidine)-1-yl-propyl]-benzooxazole;
2-[3-(4-n-butylpiperidine)-1-yl-propyl]-benzooxazole;
2-[2-(4-methylpiperidine)-1-yl-ethyl]-benzooxazole;
2-[2-(4-ethylpiperidine)-1-yl-ethyl]-benzooxazole;
2-[2-(4-n-propylpiperidine)-1-yl-ethyl]-benzooxazole;
2-[2-(4-n-butylpiperidine)-1-yl-ethyl]-benzooxazole;
2-[4-(4-methylpiperidine)-1-yl-butyl]-benzooxazole;
2-[4-(4-ethylpiperidine)-1-yl-butyl]-benzooxazole;
2-[4-(4-n-propylpiperidine)-1-yl-butyl]-benzooxazole;
2-[4-(4-n-butylpiperidine)-1-yl-butyl]-benzooxazole;
4,5-difluoro-2-(3-(4-n-butylpiperidine-1-yl)-propyl)-1H-benzoimidazole;
6-fluoro-5-nitro-2-(3-(4-n-butylpiperidine-1-yl)-propyl)-1H-benzoimidazole;
5-tert-butyl-2-(3-(4-n-butylpiperidine-1-yl)-propyl)-1H-benzoimidazole;
5-chloro-6-methyl-2-(3-(4-n-butylpiperidine-1-yl)-propyl)-1H-benzoimidazole;
4,6-difluoro-2-(3-(4-n-butylpiperidine-1-yl)-propyl)-1H-benzoimidazole;
2-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-imidazo[4,5-c]pyridine;
8-(3-(4-n-butylpiperidine)-1-yl-propyl)-9H-purine;
7-(3-(4-n-butylpiperidine)-1-yl-propyl)-3,8-dihydro-imidazo[4xe2x80x2,5xe2x80x2:3,4]benzo[1,2-d][1,2,3]triazole;
2-(3-(4-n-butylpiperidine)-1-yl-propyl)-3a,4,5,6,7,7a-hexahydro-1H-benzoimidazole;
3-methyl-1-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-indole;
5-bromo-1-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-indole;
3-formyl-1-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-indole;
7-bromo-1-(3-(4-n-butylpiperidine)-1-yl-propyl)1H-indole;
3-(3-(4-n-butylpiperidine)-1-yl-propyl)-benzo[d]isoxazole;
4-nitro-2-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-benzoimidazole;
5-nitro-2-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-benzoimidazole
4-hydroxy-2-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-benzoimidazole;
4-methyl-2-(3-(4-n-butylpiperidine)-1-yl-propyl)-1H-benzoimidazole;
3-(2-(4-n-Butylpiperidine)-ethoxy)-7-methyl-benzo[d]isoxazole;
1-(3-(4-Methylpiperidine)-1-yl-propyl)-1H-indazole;
1-(3-(4-Pentylpiperidine)-1-yl-propyl)-1H-indazole;
1-(3-(4-Propylpiperidine)-1-yl-propyl)-1Hxe2x80x94;
1-(3-(4-(3-Methyl-butyl)-piperidine)-1-yl-propyl)-1H-indazole
1-(3-(4-Pentylidene-piperidine)-1-yl-propyl)-1H-indazole;
1-(3-(4-Propylidene-piperidine)-1-yl-propyl)-1H-indazole
1-Benzo[b]thiophen-2-yl-4-(4-butylpiperidin-1-yl)-butan-1-one
4-(4-Butylpiperidin-1-yl)-1-(3-methyl-benzofuran-2-yl)-butan-1-one;
4-(4-Butylpiperidin-1-yl)-1-(5-fluoro-3-methyl-benzo[b]thiophen-2-yl)-butan-1-one;
1-Benzofuran-2-yl-4-(4-butylpiperidin-1-yl)-butan-1-one;
1-(3-Bromo-benzo[b]thiophen-2-yl)-4-(4-butylpiperidin-1-yl)-butan-1-one
1-(3-Benzo[b]thiophen-2-yl-propyl)-4-butylpiperidine;
1-(3-Benzofuran-2-yl-propyl)-4-butylpiperidine;
4-Butyl-1-[3-(3-methyl-benzofuran-2-yl)-propyl]-piperidine;
4-Butyl-1-[3-(5-fluoro-3-methyl-benzo[b]thiophen-2-yl)-propyl]-piperidine;
2-(3-Iodo-propyl)-benzo[b]thiophene;
1-(3-Benzo[b]thiophen-2-yl-propyl)-4-methylpiperidine
1-(3-Benzo[b]thiophen-2-yl-propyl)-4-benzylpiperidine;
1-(3-Benzo[b]thiophen-2-yl-propyl)-4-(2-methoxy-phenyl)-piperidine;
2-(3-Bromopropyl)-2H-benzotriazole;
2-[3-(4-Butylpiperidin-1-yl)-propyl]-2H-benzotriazole;
1-(3-Bromopropyl)-1H-benzotriazole;
1-[3-(4-Butylpiperidin-1-yl)-propyl]-1H-benzotriazole;
1-[3-(4-Butylpiperidin-1-yl)-propyl]-1H-indole-3-carbaldehyde;
{1-[3-(4-Butylpiperidin-1-yl)-propyl]-1H-indol-3-yl}-methanol;
1-[3-(4-Butylpiperidin-1-yl)-propyl]-2-phenyl-1H-benzoimidazole;
1-[3-(4-Butylpiperidin-1-yl)-propyl]-3-chloro-1H-indazole;
1-[3-(4-Butylpiperidin-1-yl)-propyl]-6-nitro-1H-indazole;
Benzo[d]isoxazol-3-ol;
3-(2-Chloroethoxy)-benzo[d]isoxazole;
3-[2-(4-Butylpiperidin-1-yl)-ethoxy]-benzo[d]isoxazol;
3-(1H-Indol-3-yl)-propan-1-ol;
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-indole hydrochloride;
4-(4-Butylpiperidine-1-yl)-butyric acid methyl ester;
2-[3-(4-Butylpiperidin-1-yl)-propyl]-1-methyl-1H-benzimidazole;
1H-Indazole-3-carboxylic acid (2-(4-butylpiperidin)-1-yl-ethyl)-amide;
1-[3-(4-Butylpiperidin-1-yl)-propyl]-5-nitro-1H-indazole;
2-[3-(4-butylpiperidin-1-yl)-propyl]-5-nitro-2H-indazole;
1-[3-(4-Butyl-piperidin-1-yl)-propyl]-2-methyl-1H-indole;
1-{1-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-indol-3-yl}-ethanone;
{1-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-indol-3-yl}-acetonitrile;
1-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-indole-3-carbonitrile;
1-[3-(4-Butyl-piperidin-1-yl)-propyl]-5,6-dimethyl-1H-benzoimidazole;
1-[3-(4-Butyl-piperidin-1-yl)-propyl]-5 (6)-dimethyl-1H-benzoimidazole;
1-[3-(4-Butyl-piperidin-1-yl)-propyl]-5-methoxy-1H-benzoimidazole;
{1-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-benzoimidazol-2-yl}-methanol;
1-[3-(4-Butyl-piperidin-1-yl)-propyl]-2-trifuoromethyl-1H-benzoimidazole;
(2-Trimethylstannanyl-phenyl)-carbamic acid tert-butyl ester;
[2-(4-Chloro-butyryl)-phenyl]-carbamic acid tert-butyl ester;
{2-[4-(4-Butyl-piperidine-1-yl)-butyryl]-phenyl}-carbamic acid tert-butyl ester;
3-[3-(4-Butyl-piperidine-1-yl)-propyl]-1H-indazole, HCl;
3-[3-(4-Butyl-piperidine-1-yl)-propyl]-5-nitro-1H-indazole;
3-[3-(4-Butyl-piperidine-1-yl)-propyl]-5,7-dinitro-1H-indazole;
4-(4-Butyl-piperidin-1-yl)-1-(2-metylsulfanyl-phenyl)-butan-1-one;
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-benzo[d]isothiazole;
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-5-methoxy-1H-indazole;
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-4-methoxy-1H-indazole
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-6-methoxy-1H-indazole;
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-indazole-4-ol(53MF51);
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-indazole-6-ol(53MF52); and
3-[3-(4-Butyl-piperidin-1-yl)-propyl]-1H-indazole-5-ol
The present invention further provides pharmaceutical compositions comprising an effective amount of at least one compound of the invention, inclusive of all compounds within the scope of formula (I).
In general, compounds of the present invention are active at cholinergic, specifically muscarinic receptors. Preferred compounds share the common property of acting as agonists at the m1 or m4 muscarinic receptor subtypes, or both. In a preferred embodiment, the compounds of the present invention are selective towards the m1, m4, or both the m1 and m4 subtypes of muscarinic receptors, i.e., the compounds have less or substantially no effect on other subtypes of the muscarinic receptors. Typically, the m1 and/or m4 selective compounds of the invention have no effect on other related receptors, including G-protein coupled receptors, e.g., serotonin, histamine, dopamine or adrenergic receptors. The invention provides compounds that are selective as agonists at either the m1 or the m4 subtype as well as compounds that are agonists at both the m1 and m4 receptor subtypes. In one embodiment, the compounds of the present invention have less or substantially no effect on m2 and m3 subtypes of muscarinic receptors. In another embodiment, the compounds of the present invention have less or substantially no effect on m2, m3, m4, and m5 subtypes of muscarinic receptors.
The compounds of present invention typically have therapeutic effects and can be used to treat or alleviate symptoms of disease conditions associated with cholinergic receptors such as cognitive impairment, forgetfulness, confusion, memory loss, attentional deficits, deficits in visual perception, depression, pain, sleep disorders, psychosis, hallucinations, aggressiveness, paranoia, and increased intraocular pressure. The disease condition may result from dysfunction, decreased activity, modification, mutation, truncation, or loss of cholinergic receptors, especially muscarinic receptors, as well as from reduced levels of acetylcholine.
The compounds of present invention can also be used to treat diseases, e.g., age-related cognitive decline, Alzheimer""s disease, Parkinson""s disease, Huntington""s chorea, Friederich""s ataxia, Gilles de la Tourette""s Syndrome, Down Syndrome, Pick disease, dementia, clinical depression, age-related cognitive decline, attention-deficit disorder, sudden infant death syndrome, and glaucoma.
The compounds of the present invention have the ability to increase cholinergic receptor activity or activate cholinergic receptors. Cholinergic receptor activity includes signaling activity or any other activity that is directly or indirectly related to cholinergic signaling or activation. The cholinergic receptors include muscarinic receptors, especially the m1 or m4 subtype of muscarinic receptors. The muscarinic receptor can be, for example, in the central nervous system, peripheral nervous system, gastrointestinal system, heart, endocrine glands, or lungs. The muscarinic receptor can be a wild-type, truncated, mutated, or modified cholinergic receptor. Kits comprising the compounds of the present invention for increasing cholinergic receptor activity or activating cholinergic receptors are also contemplated by the present invention.
The system containing the cholinergic receptor may, for example, be a subject such as a mammal, non-human primate or a human. The system may also be an in vivo or in vitro experimental model, such as a cell culture model system that expresses a cholinergic receptor, a cell-free extract thereof that contains a cholinergic receptor, or a purified receptor. Non-limiting examples of such systems are tissue culture cells expressing the receptor, or extracts or lysates thereof. Cells that may be used in the present method include any cells capable of mediating signal transduction via cholinergic receptors, expecially the m1 muscarinic receptor, either via endogenous expression of this receptor (certain types of neuronal cells lines, for example, natively express the m1 receptor), or such as following introduction of the an exogenous gene into the cell, for example, by transfection of cells with plasmids containing the receptor gene. Such cells are typically mammalian cells (or other eukaryotic cells, such as insect cells or Xenopus oocytes), because cells of lower life forms generally lack the appropriate signal transduction pathways for the present purpose. Examples of suitable cells include: the mouse fibroblast cell line NIH 3T3 (ATCC CRL 1658), which responds to transfected m1 receptors by increased growth; RAT 1 cells (Pace et al., Proc. Natl. Acad. Sci. USA 88:7031-35 (1991)); and pituitary cells (Vallar et al., Nature 330:556-58 (1987)). Other useful mammalian cells for the present method include but are not limited to HEK 293 cells, CHO cells and COS cells.
The compounds of the present invention also have the ability to reduce intraocular pressure and therefore can be used in the treatment of such diseases as glaucoma. Glaucoma is a disease in which an abnormality is observed in the circulation-control mechanism of the aqueous humor filling up the anterior chamber, i.e., the space formed between the cornea and the lens. This leads to an increase in the volume of the aqueous humor and an increase in intraocular pressure, consequently leading to visual field defects and even to loss of eyesight due to the compulsion and contraction of the papillae of the optic nerve.
The present invention also pertains to the field of predictive medicine in which pharmacogenomics is used for prognostic (predictive) purposes. Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons (see e.g., Eichelbaum, Clin Exp Pharmacol. Physiol., 23:983-985 (1996) and Linder, Clin. Chem. 43:254-66 (1997)). In general, two types of pharmacogenetic conditions can be differentiated: genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur as naturally-occurring polymorphisms.
One pharmacogenomics approach to identifying genes that predict drug response, known as xe2x80x9ca genome-wide associationxe2x80x9d, relies primarily on a high-resolution map of the human genome consisting of known gene-related markers (e.g., a xe2x80x9cbi-allelicxe2x80x9d gene marker map that consists of 60,000-100,000 polymorphic or variable sites on the human genome, each of which has two variants). Such a high-resolution genetic map can be compared to a map of the genome of each of a statistically significant number of patients taking part in a Phase II/III drug trial to identify markers associated with a particular observed drug response or side effect. Alternatively, such a high resolution map can be generated from a combination of some ten-million known single nucleotide polymorphisms (SNPs) in the human genome. As used herein, a xe2x80x9cSNPxe2x80x9d is a common alteration that occurs in a single nucleotide base in a stretch of DNA. For example, a SNP may occur once per every 1,000 bases of DNA. A SNP may be involved in a disease process although the vast majority may not be disease-associated. Given a genetic map based on the occurrence of such SNPs, individuals can be grouped into genetic categories depending on a particular pattern of SNPs in their individual genome. In such a manner, treatment regimens can be tailored to groups of genetically similar individuals, taking into account traits that may be common among such genetically similar individuals.
Alternatively, a method termed the xe2x80x9ccandidate gene approachxe2x80x9d can be utilized to identify genes that predict drug response. According to this method, if a gene that encodes a drug""s target is known (e.g., a protein or a receptor of the present invention), all common variants of that gene can be identified in the population. It can be readily determined by standard techniques a particular version of the gene is associated with a particular drug response.
Alternatively, a method termed xe2x80x9cgene expression profilingxe2x80x9d can be utilized to identify genes that predict drug response. For example, the gene expression of an animal dosed with a drug (e.g., a compound or composition of the present invention) can give an indication whether gene pathways related to toxicity have been turned on.
Information generated from more than one of the above pharmacogenomics approaches can be used to determine appropriate dosage and treatment regimens for prophylactic or therapeutic treatment of an individual. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with a compound or composition of the invention, such as a modulator identified by one of the exemplary screening assays described herein. These approaches can also be used to identify novel candidate receptor or other genes suitable for further pharmacological characterization in vitro and in vivo.
Accordingly, another aspect of the present invention features methods and kits for identifying a genetic polymorphism predisposing a subject to being responsive to a compound described herein. The method comprises administering to a subject an effective amount of a compound; identifying a responsive subject having an ameliorated disease condition associated with a cholinergic receptor; and identifying a genetic polymorphism in the responsive subject, wherein the genetic polymorphism predisposes a subject to being responsive to the compound. Identifying a genetic polymorphism in the responsive subject can be performed by any means known in the art including the methods discussed above. In addition, a kit to be used for identifying a genetic polymorphism predisposing a subject to being responsive to a compound provided in the present invention comprises the compound of the present invention, and preferably reagents and instructions for performing a genetic polymorphism test.
In one embodiment, a subject can be tested for a known polymorphism that predisposes the subject to being responsive to the compound of the present invention. The presence of the polymorphism indicates that the subject is suitable for treatment.
In preferred embodiments, the compounds of the present invention can be represented as shown in formulae (IIIa-e): 
where W1 is O, S, or NR5, W2 is CR5 or N, and W3 is CR5 or N, or
wherein W3 is NR5, S or O,
or a pharmaceutically acceptable salt, ester or prodrug thereof.
Compounds of the present invention may be prepared by methods analogous to the methods disclosed in G.B. Patent No. 1,142,143 and U.S. Pat. No. 3,816,433, each of which are incorporated herein by reference. Ways of modifying those methods to include other reagents etc. will be apparent to those skilled in the art. Thus, for instance, compounds of formula (III, e.g., IIIb where W1 is NR5) may be prepared as shown in the following reaction scheme. 
The starting compound having formula (X) may be prepared by general methods of organic synthesis. For general methods of preparing compounds of formula (X), reference is made to Fuller, et al., J. Med. Chem. 14:322-325 (1971); Foye, et al., J. Pharm. Sci. 68:591-595 (1979); Bossier, et al., Chem. Abstr. 66:46195h and 67:21527a (1967); Aldous, J. Med. Chem. 17:1100-1111 (1974); Fuller, et al., J. Pharm. Pharmacol. 25:828-829 (1973); Fuller, et al., Neuropharmacology 14:739-746 (1975); Conde, et al., J. Med. Chem. 21:978-981 (1978); Lukovits, et al., Int. J. Quantum Chem. 20:429-438 (1981); and Law, Cromatog. 407:1-18 (1987), the disclosures of which are incorporated by reference herein in their entirety. Compounds of formula XI are prepared, for example, as described in Darbre, et al., Helv. Chim. Acta, 67:1040-1052 (1984) or Ihara, et al., Heterocycles, 20:421-424 (1983), also incorporated herein by reference. The radiolabelled derivatives having formula (XX) may be prepared by, for example, using a tritiated reducing agent to form the reductive amination or by utilizing a 14C-labelled starting material.
Compounds of formula (XXII) can be used to prepare the compounds of formula (I). Compounds of formula (XXII) are prepared, for example, as described in Ishii, et al., J. Org. Chem. 61:3088-3092 (1996) or Britton, et al. Bioorg. Med. Chem. Lett. 9:475-480 (1999), also incorporated herein by reference. Where the starting compound includes a carbonyl group, the compound having the formula (XXII) may be reduced with, for example, AlH3, diborane:methyl sulfide or other standard carbonyl reducing reagents to produce the ligand having the formula (XXX). 
The receptor ligands having formula (XXXII) may be prepared by nucleophilic displacement of a suitable nucleophuge (E) by the amino derivative (XXXI). Examples of nucleophuges, which may be used for this purpose, include halides such as I, Cl, Br, or tosylate or mesylate. 
When Y in formula (XXX) is xe2x80x94C(O)xe2x80x94, this compound may be prepared from oxidation of a secondary alcohol with, for example, pyridinium chlorochromate, N-chlorosuccinimide, CrO3xe2x80x94H2SO4, or via the Swern or Dess-Martin proceduresxe2x80x94nickel.
When Y in formula (XXX) is xe2x80x94Oxe2x80x94, this compound may be prepared by arylation of an alcohol with arylhalides under, for example, Cu catalysis.
When Y in formula (XXX) is xe2x80x94Sxe2x80x94, this compound may be prepared by arylation of a thiol with arylhalides under, for example, Cu catalysis.
When Y in formula (XXX) is xe2x80x94CHOHxe2x80x94, this compound may be prepared by reduction of the corresponding ketone by catalytic hydrogenation or by the use of NaBH4 or by the use of LiAlH4.
Suitable pharmaceutically acceptable salts of the compounds of this invention include acid addition salts which may, for example, be formed by mixing a solution of the compound according to the invention with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulphuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Furthermore, where the compounds of the invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, e.g., sodium or potassium salts; alkaline earth metal salts, e.g., calcium or magnesium salts; and salts formed with suitable organic ligands, e.g., quaternary ammonium salts. Examples of pharmaceutically acceptable salts include the acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium, carbonate, chloride, clavulanate, citrate, dihydrochloride, fumarate, gluconate, glutamate, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, nitrate, N-methylglucamine ammonium sal, oleate, oxalate, phosphate/diphosphate, salicylate, stearate, sulfate, succinate, tannate, tartrate, tosylate, triethiodide and valerate salt.
The present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs are derivatives of the compounds of this invention, which are readily convertible in vivo into the required compound. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in Design of Prodrugs, (Bundgaard, ed. Elsevier, 1985). Metabolites of these compounds include active species produced upon introduction of compounds of this invention into the biological milieu.
Where the compounds according to the invention have at least one chiral center, they may exist as a racemate or as enantiomers. It should be noted that all such isomers and mixtures thereof are included in the scope of the present invention. Furthermore, some of the crystalline forms for compounds of the present invention may exist as polymorphs and as such are intended to be included in the present invention. In addition, some of the compounds of the present invention may form solvates with water (i.e., hydrates) or common organic solvents. Such solvates are also included in the scope of this invention.
Where the processes for the preparation of the compounds according to the invention give rise to mixtures of stereoisomers, such isomers may be separated by conventional techniques such as preparative chiral chromatography. The compounds may be prepared in racemic form, or individual enantiomers may be prepared either by stereoselective synthesis or by resolution. The compounds may, for example, be resolved into their component enantiomers by standard techniques, such as the formation of diastereomeric pairs by salt formation with an optically active acid, such as (xe2x88x92)-di-p-toluoyl-d-tartaric acid and/or (+)-di-p-toluoyl-1-tartaric acid followed by fractional crystallization and regeneration of the free base. The compounds may also be resolved by formation of diastereomeric esters or amides, followed by chromatographic separation and removal of the chiral auxiliary.
During any of the processes for preparation of the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry (McOmie ed., Plenum Press, 1973); and Greene and Wuts, Protective Groups in Organic Synthesis (John Wiley and Sons, 1991) The protecting groups may be removed at a convenient subsequent stage using methods known from the art.
Compounds of the present invention may be administered in any of the foregoing compositions and according to dosage regimens established in the art whenever specific pharmacological modification of the activity of muscarinic receptors is required.
The present invention also provides pharmaceutical compositions comprising one or more compounds of the invention together with a pharmaceutically acceptable diluent or excipient. Preferably such compositions are in unit dosage forms such as tablets, pills, capsules (including sustained-release or delayed-release formulations), powders, granules, elixirs, tinctures, syrups and emulsions, sterile parenteral solutions or suspensions, aerosol or liquid sprays, drops, ampoules, auto-injector devices or suppositories; for oral, parenteral (e.g., intravenous, intramuscular or subcutaneous), intranasal, sublingual or rectal administration, or for administration by inhalation or insufflation, and may be formulated in an appropriate manner and in accordance with accepted practices such as those disclosed in Remington ""s Pharmaceutical Sciences (Gennaro, ed., Mack Publishing Co., Easton Pa., 1990). Alternatively, the compositions may be in sustained-release form suitable for once-weekly or once-monthly administration; for example, an insoluble salt of the active compound, such as the decanoate salt, may be adapted to provide a depot preparation for intramuscular injection. The present invention also contemplates providing suitable topical formulations for administration to, e.g., eye, skin or mucosa.
For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, flavoring agents and coloring agents can also be incorporated into the mixture. Suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or beta-lactose, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes and the like. Lubricants used in these dosage forms include, without limitation, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
For preparing solid compositions such as tablets, the active ingredient is mixed with a suitable pharmaceutical excipient, e.g., such as the ones described above, and other pharmaceutical diluents, e.g., water, to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention, or a pharmaceutically acceptable salt thereof. By the term xe2x80x9chomogeneousxe2x80x9d is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective unit dosage forms such as tablets, pills and capsules. The solid preformulation composition may then be subdivided into unit dosage forms of the type described above containing from about 0.01 to about 50 mg of the active ingredient of the present invention. The tablets or pills of the present composition may be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner core containing the active compound and an outer layer as a coating surrounding the core. The outer coating may be an enteric layer, which serves to resist disintegration in the stomach and permits the inner core to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, such materials including a number of polymeric acids and mixtures of polymeric acids with conventional materials such as shellac, cetyl alcohol and cellulose acetate.
The liquid forms in which the present compositions may be incorporated for administration orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oil suspensions, and flavored emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar pharmaceutical carriers. Suitable dispersing or suspending agents for aqueous suspensions include synthetic and natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, gelatin, methylcellulose or polyvinyl-pyrrolidone. Other dispersing agents, which may be employed, include glycerin and the like. For parenteral administration, sterile suspensions and solutions are desired. Isotonic preparations, which generally contain suitable preservatives, are employed when intravenous administration is desired. The compositions can also be formulated as an ophthalmic solution or suspension formation, i.e., eye drops, for ocular administration.
Compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses two, three or four times daily. Furthermore, compounds of the present invention may be administered in intranasal form via topical use of suitable intranasal vehicles or via transdermal routes, using e.g., forms of transdermal skin patches that are well known to persons skilled in the art. To be administered in the form of a transdermal delivery system, the dosage administration will be continuous rather than intermittent throughout the dosage regimen.
The dosage regimen utilizing the compounds of the present invention is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound employed. A physician or veterinarian of ordinary skill can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the disease or disorder, which is being treated.
The daily dosage of the products may be varied over a wide range from 0.01 to 100 mg per adult human per day. For oral administration, the compositions are preferably provided in the form of tablets containing 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0 or 50.0 mg of the active ingredient for the symptomatic adjustment of the dosage to the patient to be treated. A unit dose typically contains from about 0.001 mg to about 50 mg of the active ingredient, preferably from about 1 mg to about 10 mg of active ingredient. An effective amount of the drug is ordinarily supplied at a dosage level of from about 0.0001 mg/kg to about 25 mg/kg of body weight per day. Preferably, the range is from about 0.001 to 10 mg/kg of body weight per day, and especially from about 0.001 mg/kg to 1 mg/kg of body weight per day. The compounds may be administered, for example, on a regimen of 1 to 4 times per day.
Compounds according to the present invention may be used alone at appropriate dosages defined by routine testing in order to obtain optimal pharmacological effect on a muscarinic receptor, in particular the muscarinic m1 or m4 receptor subtype, while minimizing any potential toxic or otherwise unwanted effects. In addition, co-administration or sequential administration of other agents that improve the effect of the compound may, in some cases, be desirable.
The pharmacological properties and the selectivity of the compounds of this invention for specific muscarinic receptor subtypes may be demonstrated by a number of different assay methods using, for example, recombinant receptor subtypes, preferably of the human receptors as available, e.g., conventional second messenger or binding assays. A particularly convenient functional assay system is the receptor selection and amplification assay disclosed in U.S. Pat. No. 5,707,798, which describes a method of screening for bioactive compounds by utilizing the ability of cells transfected with receptor DNA, e.g., coding for the different muscarinic subtypes, to amplify in the presence of a ligand of the receptor. Cell amplification is detected as increased levels of a marker also expressed by the cells.
The invention is disclosed in further detail in the following examples, which are not in any way intended to limit the scope of the invention as claimed.