KR20010042248A - Compounds with activity on muscarinic receptors - Google Patents

Abstract

The present invention relates to compounds having activity against muscarinic receptors,

Modifying mucarrin m1 receptor activity by the above compounds and methods has a beneficial effect and is capable of alleviating or treating a disease or disorder in which a therapeutically effective amount of a selective muscarinic m1 functional compound is capable of modulating Which is administered to a patient.

Description

[0002] COMPOUNDS WITH ACTIVITY ON MUSCARINIC RECEPTORS [0003]

Muscarinic acetylcholine receptors play a central role in the central nervous system for high cognitive function as well as in the peripheral parasympathetic nervous system. Cloning yields five distinct muscarinic receptor subtypes (i. E., M1-m5) (TI Bonner et al., Science 237, 1987, pp. 527-532; TI Bonner et al., Neuron 1, 1988, pp. 403-410). m1 is a subtype of dominance in the cerebral cortex and is believed to be involved in controlling cognitive function, m2 is dominant in the heart and is believed to be involved in controlling heart rate, while m3 is associated with sweating and salivation It is also believed to be involved in the gastrointestinal and urinary tract stimulation, and that m4 is present in the brain and m5 is present in the brain and is involved in the central function of the central nervous system associated with the dopaminergic system.

Animal studies of several muscarinic ligands (S. Iversen, Life Sciences 60 (Nos. 13/14), 1997, pp. 1145-1152) have shown that muscarinic compounds have a profound effect on cognitive functions such as learning and memory . This suggests the potential use of muscarinic agonists to improve cognitive function in age-related (such as Alzheimer's or other dementia) and age-independent (eg, attention deficit hyperactivity disorder) cognitive impairment.

Based on the presence of the muscarinic receptor subtype in various tissues, it has been found that the m1 receptor subtype is a more abundant subtype in the cerebral cortex, cerebral basal ganglia and hippocampus, accounting for 35-60% of all muscarinic receptor binding sites (A. Levey, Proc Natl Acad Sci USA 93, 1996, pp. 13541-13546). This suggests that the m1 (and possibly m4) subtype plays a major role as a synaptic postmusclein receptor (located in the neocortex and acetylcholine receptive neurons in hippocampus) in a number of cognitive and motor functions, m1 response.

It has been found that diseases associated with cognitive impairment such as Alzheimer's disease are established by the selective loss of acetylcholine in the brain. It is believed that the neocortical domain of the cortex and hippocampus denatures cholinergic neurons in the basal ganglia involved in advanced functions (see S. Iversen, supra). The discovery suggests that the diseases are treated or at least restored by a drug that enhances cholinergic function in the infected area of the brain.

(AChE) inhibitor, such as 9-amino-1,2,3,4-tetrahydroacridine (tacrine), to inhibit the brain's acetylcholine, which indirectly stimulates muscarinic receptors, . Tacrine therapy improves cognition adequately and temporarily in Alzheimer's patients (Kasa et al., Supra). On the other hand, tacrine has been found to have cholinergic side effects due to peripheral acetylcholine stimulation. These include abdominal cramps, nausea, vomiting, diarrhea, poor appetite, weight loss, muscle warts, and heartburn. Gastrointestinal adverse events were found in about one-third of patients treated. Tacrine has also been found to cause severe hepatotoxicity, increased hepatic aminotransferase, found in about 30% of patients (P. Taylor, " Anticholinergic Agents ", Chapter 8 in Goodman and Gilman: The Pharmacological Basis of Therapeutics , 9th Edition, 1996, pp. 161-176). Side effects of tacrine severely limit his clinical application. (R, S) -1-benzyl-4- [5,6-dimethoxy-1-indanone-2-yl] methylpiperidine. HCl (Donepezil) Has recently been shown to alleviate Alzheimer's disease (see P. Kasa et al., Supra). The compound has no gastrointestinal effects similar to the effect of tacrine, which stimulates the m3 receptor caused by elevated parasympathetic nerve activity levels, although no liver damage has been found.

Since the muscarinic m1 receptor in the prefrontal cortex and the hippocampus appears to be an intact disease, it has already been shown that administering a drug acting as an agonist to the muscarinic receptor can treat or at least alleviate acetylcholine loss in Alzheimer's patients (JH Brown and P. Taylor, " Muscarinic Receptor Agonists and Antagonists ", Chapter 7 in Goodman and Gilman: The Pharmacological Basis of Therapeutics, 9th Edition, 1996, p.147).

Muscarinic agonists (believed to be m1 selective) presented here for treating Alzheimer's disease such as arecolin did not appear to be more efficacious in clinical trials than AChE inhibitors (see S. V. P. Jones et al., Supra). In one study (T. Sunderland et al., Brain Res. Rev. 13, 1988, pp. 371-389), arecoline significantly increased motor activity, significantly elevated the atmosphere, significantly reduced anergia As a result of the change in behavior that is often found in Alzheimer's disease patients. However, the putative m1 agonist was later found to be a weak partial agonist selective for m2 and / or m3 receptor subtypes (H. Brauner-Osborne et al., J. Med. Chem. 38, 1995, pp.2188- 2195). As noted above, m2 subtype selectivity is presumed to be responsible for the cardiovascular effects found for the agonists, such as tachycardia and bradycardia, and m3 action is believed to be responsible for gastrointestinal side effects of the agonist.

Therefore, the m2 and / or m3 action can be used to treat severe Alzheimer ' s disease by severely limiting the dose of drug that can be administered to patients who have received suboptimal dosages so far, This is a disadvantage. And the lack of subtype selectivity and low latency of currently tested cholinergic compounds show negative peripheral side effects and have limited cognitive effects due to weak and / or counteracting actions in the brain. Thus, it is very advantageous to develop compounds with improved selectivity for the m1 subtype, with little or no effect on the m2 and m3 subtypes.

SUMMARY OF THE INVENTION

The present invention provides a compound having muscarinic agonist activity of the formula (I): EMI5.1 or a pharmaceutically acceptable salt, ester or prodrug thereof.

(I)

X ₁ , X ₂ , X ₃ , X ₄ and X ₅ are selected from C, N and O;

k is 0 or 1;

t is 0, 1 or 2;

R ₁ is a linear or branched substituted with C _1-3 alkyl or halo-chain C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _1-8 alkylidene, C _1-8 alkoxy , C _1-8 heteroalkyl, C _1-8 aminoalkyl, C _1-8 haloalkyl, C _1-8 alkoxycarbonyl, C _1-8 hydroxy alkoxy, C _1-8 hydroxyalkyl, -SH, C _1-8 alkylthio, -O-CH ₂ -C _5-6 aryl, -C (O) -C _5-6 aryl; C _5-6 aryl or C _5-6 cycloalkyl optionally comprising one or more heteroatoms selected from N, S and O; _{-C (O) NR 3 R 4} , -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -CR 3 R 4, -OC (O) R 3, - (O) (CH 2) _s NR ₃ R ₄ or - (CH ₂ ) _s NR ₃ R ₄ ; Wherein R ₃ , R ₄ and R ₅ are the same or different and each is selected from H, C _1-6 alkyl; C _5-6 aryl is optionally composed of one or more heteroatoms selected from N, O and S, and is optionally substituted by halo or C _1-6 alkyl; C _3-6 cycloalkyl; Or R ₃ and R ₄ , when present together with the N atom, form a cyclic ring structure consisting of 5-6 atoms selected from C, N, S and O;

A is C _5-12 aryl or a C _5-7 cycloalkyl, N, S, and O, and each selectively configured as one or more selected from a heteroatom;

R ₂ is H, amino, hydroxyl, halo, or a straight or branched-chain C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 heteroalkyl , C _1-6 aminoalkyl, C _1-6 haloalkyl, C _1-6 alkylthio, C _1-6 alkoxycarbonyl, -CN, -CF ₃ , -OR ₃ , NO ₂ , -NHR ₃ , -NHC (O) R ₃ -, -C (O) NR ₃ R ₄ , -NR ₃ R ₄ , -NR ₃ C (O) NR ₄ R ₅ , -OC (O) R ₃ . _{-C (O) R 3 R 4} , -O (CH 2) q NR 3, -CNR 3 R 4 or _{- (CH 2) q NR 3} R 4 , and; Wherein q is an integer from 1 to 6;

n is 0, 1, 2, 3 or 4; when n> 1, the groups R ₂ are the same or different;

p is 0 or an integer from 1 to 5;

O, S, CHOH, -NHC Y is (O) -, -C (O ) NH-, -C (O) -, -OC (O) -, NR 7 or a -CH = N-, R ₇ is H or C _1-4 alkyl; Or is absent; And

Z is CR ₈ R ₉ , wherein R ₈ and R ₉ are each selected from H and linear or branched chain C _1-8 alkyl,

The present invention further provides a pharmaceutical composition comprising an effective amount of a compound of formula (I).

There is additionally provided a method of treating a symptom of a disease or disorder associated with a reduced level of acetylcholine, comprising administering a therapeutically effective amount of a composition comprising a compound of formula (I).

In a further embodiment, the invention further provides a method of treating a symptom of a disease or disorder associated with increased intraocular pressure, such as glaucoma, comprising administering a therapeutically effective amount of a compound of formula (I) Lt; / RTI >

The present invention relates to a method for activating muscarinic receptors and a method for treating or ameliorating diseases beneficially by modifying muscarinic receptor activity, as well as compounds selective for muscarinic acetylcholine receptor subtypes.

Figure 1 is a graph showing the raw data for screening 35,000 small organic molecules from one 96-well microtitre plate in the assay described in Example XVI,

Figure 2 is a graph showing data comparing the profiles of reference antagonist atropine with m1 muscarinic receptor infected cells stimulated by either carbachol (open triangle) or Compound A (Example I) (closed triangles).

The present invention contemplates the use of the m1 receptor subtype as compared to other muscarinic subtypes having beneficial effects in treating cognitive impairment such as Alzheimer ' s disease, avoiding the side effects of the drugs presented herein for this purpose, ≪ RTI ID = 0.0 > and / or < / RTI > Compounds exhibiting this property are isolated by screening for the m1-m5 receptor subtype.

According to one embodiment, the present invention provides a compound of formula (I), a pharmaceutically acceptable salt, ester or prodrug thereof:

In the above formula (I)

X ₁ , X ₂ , X ₃ , X ₄ and X ₅ are C; Or one of X ₁ , X ₂ , X ₃ , X ₄ or X ₅ is O or N and the other is C;

k is 0 or 1;

t is 1;

R ₁ is straight or branched C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _1-8 alkylidene, C _1-8 alkoxy, C _1-8 aminoalkyl, C _{1- 8} haloalkyl, C _{1-8 alkoxycarbonyl, -C (O) NR 3 R} 4, -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -OC (O) R 3 or - ( CH ₂ ) _s NR ₃ R ₄ wherein R ₃ , R ₄ and R ₅ are the same or different and are each selected from H and C _1-6 alkyl; s is an integer from 1 to 8;

n is 1, 2 or 3; And

A is phenyl or naphthyl;

R ₂ is straight or branched C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _{1- 6 alkoxycarbonyl, -CN, -CF 3, -OH,} -COR 3, -NHR 3, -NHC (O) R 3, -C (O) NR 3 R 4, -NR 3 R 4, -NR 3 C (O) NR ₄ R ₅ , -OC (O) R ₃ or - (CH ₂ ) _q NR ₃ R ₄ wherein q is an integer from 1 to 6; or

A is aryl containing at least one heteroatom selected from N, S and O;

R ₂ is H, halo or a straight-chain or branched C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 heteroalkyl, C _1-6 aminoalkyl , C _1-6 haloalkyl, C _{1-6 alkoxycarbonyl, -CN, -CF 3, -OH,} -COR 3, -NHR 3, -NHC (O) R 3, -C (O) NR 3 R ₄ , -NR ₃ R ₄ , -NR ₃ C (O) NR ₄ R ₅ , -OC (O) R ₃ or - (CH ₂ ) _q NR ₃ R ₄

In one preferred embodiment, the compound is a compound of the formula (II)

Preferred embodiments of the compounds of formula (II) include compounds of formula (IIa) and (IIb)

According to one preferred series of embodiments of compounds of formulas I, II, IIa and IIb, t is 1 and Y is -C (O) -, -NHC (O) -, S, O or -C to be. In another, X ₃ is C. And R ₁ is preferably alkyl, in which R ₂ is preferably alkyl, amino, alkyl, alkoxy or hydroxyl jitsu. In one embodiment, p is 3. In another, R ₁ is C _2-8 alkyl and R ₂ is methyl, hydroxyl, or alkoxy.

In one embodiment, n is 1 or 2; Y is -C (O) - or O, and t is 1. R ₂ is preferably halo. According to another embodiment, t is O; Or R < ₁ > is alkoxy, benzyl or phenyl.

X ₃ is also N, wherein according to one embodiment R ₁ is alkyl or alkoxy; Or R < ₁ > is benzyl or phenyl; Wherein R < ₂ > is alkyl or alkoxy.

According to another embodiment, X < ₃ > is O, wherein t may be, for example, O. Preferably, R < ₂ > is alkyl or alkoxy; Or R < ₂ > is halo.

Particular embodiments of the invention include:

4-methoxy-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-ethoxy-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-propoxy-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-butoxy-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-methoxymethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-ethoxymethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-propoxymethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4- (2-methoxyethyl) -1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4- (2-ethoxyethyl) -1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-methoxy-4-methyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-methoxy-4-ethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-methoxy-4-propyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-methoxy-4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-Ethoxy-4-methyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-Ethoxy-4-ethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-Ethoxy-4-propyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-ethoxy-4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-propoxy-4-methyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-propoxy-4-ethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-propoxy-4-propyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-propoxy-4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-Butoxy-4-methyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-Butoxy-4-ethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-Butoxy-4-propyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-Butoxy-4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

2- [3- (4-n-butylpiperidine) propoxy] toluene;

2- [3- (4-n-butylpiperidine) propanesulfanyl] toluene;

2- [3- (4-n-butylpiperidine) propanesulfinyl] toluene;

3- (4-n-butylpiperidine) -o-tolyl-butane-l-thione;

3- (4-n-butylpiperidinopropyl) -o-tolyl-amine;

N- (4- (4-n-butylpiperidine) -l-o-tolyl-butyl) -hydroxylamine;

4-n-butyl-1- [4- (2-chlorophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-bromophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-fluorophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-mercaptophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-sulfanylmethylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-sulfanylethylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-aminophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-methylaminophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-ethylaminophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-dimethylaminophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-diethylaminophenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (1-H-imidazol-2-yl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (1-imidazol-1-yl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (1 -thiazol-2-yl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4 - ([1,2,3] triazol-1-yl) -4-oxo-1-butyl] piperidine;

2- [4-n-Butyl-piperidin-l-ethyl] -8-methyl-3,4-dihydro-2H-naphthalen-l-one;

2- [4-n-Butyl-piperidin-l-ethyl] -7-methyl-indan-l-one;

3- [4-n-Butyl-piperidin-l-ethyl] -chroman-4-one;

2- [4-n-Butyl-piperidin-l-ethyl] -lH-benzoimidazole;

4-n-butyl-1- [4- (4-fluoro-2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-hydroxyphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-methoxyphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (1-thiophen-2-yl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-ethylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-ethoxyphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2,4-dimethylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2,3-dimethylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (3-methoxyphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-benzyloxyphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (4-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-N-phenyl-butyramide;

4-methyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (naphthalen-1-yl) -4-oxo-1-butyl] piperidine;

4-benzyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

1- [4- (2-methylphenyl) -4-oxo-1-butyl] pyrrolidine;

4-benzyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine;

2-propyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

2-ethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-n-propyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine;

3,5-dimethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-methyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine;

4-n-hexyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine;

4-hydroxyethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine;

4-ethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine;

4-benzyl-1- [4- (4-fluorophenyl) -4-oxo-1-butyl] piperidine;

4-benzyl-1- [4- (4-bromophenyl) -4-oxo-1-butyl] piperidine;

4-phenyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine;

3-hydroxymethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-methyl-1- [4- (4-bromophenyl) -4-oxo-1-butyl] piperidine;

1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

2-hydroxymethyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

4-benzyl-1- [4- (2-methylphenyl) -4-oxo-1-pentyl] piperazine;

4-n-hexyl-1- [4- (2-methylphenyl) -4-oxo-1-pentyl] piperazine;

4- (Piperidin-1-yl) -1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine;

1- [4- (2-methylphenyl) -4-oxo-1-butyl] -2,3-dihydro-1H-indole;

4-benzyl-1- [5- (2-methylphenyl) -5-oxo-1-pentyl] piperidine;

4-n-butyl-1- [5- (2-methylphenyl) -5-oxo-1-pentyl] piperidine;

4-n-butyl-1- [4- (2,6-dimethylphenyl) -4-oxo-1-butyl] piperidine;

4-n-butyl-1- [4- (2-methoxymethylphenyl) -4-oxo-1-butyl] piperidine;

1- (2-methylphenyl) -2- (4-benzylpiperazin-1-yl) -ethanone;

3,5-dimethyl-1- [5- (2-methylphenyl) -5-oxo-1-pentyl] piperidine;

3,5-dimethyl-1- [4- (4-fluorophenyl) -4-oxo-1-butyl] piperidine;

1- [4- (4-fluorophenyl) -4-oxo-1-butyl] pyrrolidine;

4-benzyl-1- [6- (2-methylphenyl) -6-oxo-1-hexyl] piperazine;

3,5-dimethyl-1- [6- (2-methylphenyl) -6-oxo-1-butyl] piperidine;

4-benzyl-1- [5- (2-methoxyphenyl) -5-oxo-1-pentyl] piperazine;

4-benzyl-l- [3-phenyl-3-oxo-l-propyl] piperazine;

4-n-butyl-1- [5- (2-methoxyphenyl) -5-oxo-1-pentyl] piperidine;

3,5-dimethyl-1- [4- (4-fluoro-2-methylphenyl) -4-oxo-1-butyl] piperidine;

3-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] azetidine;

4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-2-methyl-1-butyl] piperidine;

4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-2,2-dimethyl-1-butyl] piperidine;

4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-2-ethyl-1-butyl] piperidine;

4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-2-propyl-butyl] piperidine; And

Butyl-1- [4- (2-methylphenyl) -4-oxo-2,2-diethyl-1-butyl] piperidine

.

The compounds originally excluded in the range of formula (I) are 4-n-butyl-1- [4-phenyl-4-oxo-1-butyl] piperidine; 4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine; 2- [3- (3-n-butylpiperidine) propanesulfanyl] toluene; (I.e., - (CH ₂ ) _p -Y- is - (CH ₂ ) ₃ -) - 4-propyloxy- C (O) - or - (CH ₂ ) ₃ -S-, X ₁ to X ₅ are C, -A- (R ₂ ) _n and R ₁ are not together: o- -Butyl, respectively, phenyl and n-butyl, or p-fluoro-phenyl and -O- (CH ₂ ) ₂ CH ₃ , respectively.

The invention also relates to the use of a compound of formula (I) (i.e., 4-n-butyl-1- [4-phenyl-4-oxo-1-butyl] piperidine 2- [3- (3-n-butylpiperidine) propanesulfanyl] toluene; 4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine; And an agonizing method of muscarinic receptors comprising reacting a receptor with a 4-propyloxy-1- [4- (4-fluorophenyl) -4-oxo-1-butyl] piperidine .

The present invention also relates to a pharmaceutical composition comprising an effective amount of a compound of formula (I) (i.e., -n-butyl-1- [4-phenyl-4-oxo-1-butyl] piperidine; 2- [3- (3-n-butylpiperidine) propanesulfanyl] toluene, and 4- [4- Propoxy-1- [4- (4-fluorophenyl) -4-oxo-1-butyl] piperidine).

The present invention also provides a method of treating a symptom of a disease or disorder associated with a reduced level of acetylcholine, a method comprising administering a therapeutically effective amount of a composition disclosed herein. Examples of diseases or disorders include neurogenic human disease, cognitive impairment, age-related cognitive decline, or dementia.

The compounds of the present invention can also be used to treat diseases such as glaucoma since their ability to reduce intraocular pressure has been demonstrated. Glaucoma is a disease in which abnormality is found in the circulatory-control mechanism of aqueous body fluid filling the anterior chamber, which is the space formed between the cornea and the lens. This increases the volume of the aqueous fluid, increases the intraocular pressure, eventually narrowing the field of vision, and causing visual loss due to forcing and reduction of the optic nerve head.

Preferably, the compounds of the invention show selective agonist activity on the m1 receptor. The agent is defined as a compound that increases the activity of the m1 muscarinic receptor when contacted with the receptor. Selectivity is defined as the property of the muscarinic m1 agonist that the amount of agonist effective in increasing the activity of the m1 receptor does not substantially increase or increase the activity of the m3 and m5 subtypes and preferably the m2 and m4 subtypes.

As used herein, the term "alkyl" refers to straight or branched chain alkane groups having 1-6 carbon atoms in the chain, such as methyl, ethyl, propyl, isopropyl, n-butyl, sec- . The term " heteroalkyl " is considered to refer to an alkane group containing one or two heteroatoms selected from O, S or N.

As used above, the term " alkenyl " means a straight chain or branched chain alkene group having 2-6 carbon atoms in the chain; The term " alkynyl " is considered to refer to a straight or branched-chain alkyne group having 2-6 carbon atoms in the chain.

The term " aryl " and " cycloalkyl ", as used hereinbefore, preferably denotes a single ring and bicyclic ring structure of 5 to 12 carbon atoms, more preferably a monocyclic ring of 5 to 6 carbon atoms . When the ring is composed of one or more heteroatoms selected from N, S and O (i.e., a heterocyclic ring), the ring consists of a total of 5 to 12 atoms, more preferably 5 to 6 atoms. The heterocyclic ring may be optionally substituted with one or more substituents selected from the group consisting of furyl, pyrroyl, pyrazolyl, thienyl, imidazolyl, isoxazolyl, oxazolyl, thiazolyl, isothiazolyl, pyridyl, piperidinyl, Thiadiazolyl, imidazolinyl, imidazolidinyl, and the like, but are not limited thereto. The ring may be substituted by one or more groups included in the definition of R < ₂ >. Substituted C _1-6 alkyl, C _1-6 alkenyl, C _1-6 alkynyl, C _1-6 alkoxy, C _1-6 heteroalkyl, C _1-6 aminoalkyl, C _1-6 haloalkyl, or C _{1 -6} alkoxycarbonyl is understood to be substituted by one or more hydroxyl, C _1-4 alkoxy, halogen, cyano, amino or nitro if present.

As used herein, the terms "halogen" or "halo" include chlorine, fluorine, iodine, and bromine.

It is understood that all of the rings represented by the structures below can be saturated and unsaturated:

The compounds of the present invention may be prepared by methods analogous to those described in British Patent GB 1,142,143 and US Patent 3,816,433. Methods of modifying the above methods, including other reactive agents and the like, will be apparent to those skilled in the art. Thus, for example, compounds of formula (I) may be prepared as shown in Scheme 1 below.

The starting compounds of formula (X) can be prepared by conventional organic synthesis methods. General methods for preparing compounds of formula (X) are described in Fuller, RW et al., J. Med. Chem. 14: 322-325 (1971); Foye, WO et al., J. Pharm. Sci. 68: 591-595 (1979); Bossier, JR et al., Chem. Abstr. 66: 46195h and 67: 21527a (1967); Aldous, FAB, J. Med. Chem. Abstr. 17: 1100-1111 (1974); Fuller, RW et al., J. Pharm. Pharmacol. 25: 828-829 (1973); Fuller, RW et al., Neuropharmacology 14: 739-746 (1975); Conde, S et al., J. Med. Chem. 21: 978-981 (1978); Lukovits, I. et al., Int. J. Quantum Chem. 20: 429-438 (1981); And Law, B., J. Cromatog. 407: 1-18 (1987), the text of which is incorporated herein by reference. The radiolabeled derivative of formula (XX) may be prepared using a trichloro reductant to achieve reductive amination or using a ¹⁴ C-labeled starting material.

Alternatively, the starting compound comprises a carbonyl group, and the compound of formula (XXII) is reduced, for example, by AlH ₃ , diborane: methyl sulfide or other standard carbonyl reducing agent to form a ligand of formula (XXX).

Receptor ligands of the formula (XXXII) can be prepared by nucleophilic substitution of the electrophiles (E) with an amino derivative (XXXI). Examples of electrophiles which may be used for this purpose include halides such as I, Cl, Br, tosylate or mesylate.

The Y in the general formula (ⅩⅩⅩII) -C (O) - . For, the compounds sec flutes alcohol pyridinium chloro chromate or N- chloro succinimide or a CrO ₃ -H ₂ SO ₄ or nickel peroxide or metal (Al , ≪ / RTI > K) or DCC-DMSO.

When Y is -O- in formula (XXXII), the compound may be prepared by alkylation of an alcohol with an aryl halide, for example under Cu catalysis.

When Y is -S- in formula (XXXII), the compound may be prepared, for example, by alkylation of the thiol with an aryl halide under Cu catalysis.

If the formula Y is -CHOH- in (ⅩⅩⅩII), the compounds can be prepared by the use of reduced or NaBH ₄ or LiAlH ₄ using the corresponding ketone by catalytic hydrogenation.

Suitable pharmaceutically acceptable salts of the compounds of the present invention include the pharmaceutically acceptable acid addition salts such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, oxalic acid, citric acid, tartaric acid, ≪ / RTI > And, where the compound of the present invention carries an acid component, its suitable pharmaceutically acceptable salts include alkali metal salts such as sodium EH, potassium salt; Alkaline earth metal salts such as calcium or magnesium salts; And salts formed with suitable organic ligands such as quaternary ammonium salts. Examples of pharmaceutically acceptable salts include, but are not limited to, 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, methyl bromide, methyl nitrate , Methyl sulfate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate, phosphate / diphosphate, salicylate, stearate, sulfate, succinate, tannate, tartrate, tosylate, And valerate salts.

The present invention includes within its scope prodrugs of the compounds of the present invention. Usually, the prodrugs are inactive derivatives of the compounds of the present invention which are readily convertible to the required compound in vivo. Conventional methods for the selection and preparation of suitable prodrug derivatives are described, for example, in " Design of Prodrugs " ed. H. Bundgaard, Elsevier, 1985. Metabolites of such compounds include those active substances that are produced when the compounds of the present invention are introduced into a biological environment.

Where the compounds according to the invention have at least one chiral center, they may be present as racemates or as optical isomers. It is to be understood that all such isomers and mixtures thereof are included within the scope of the present invention. And, some of the crystal forms for the compound of the present invention may exist as a homogeneous isomer, and are considered to be included in the present invention as described above. And some of the compounds of the present invention form a solvent or a conventional organic solvent having water (i.e., hydrate). Such solvent compounds are also included within the scope of the present invention.

Where the method for preparing the compounds according to the invention forms a mixture of stereoisomers, the isomers may be separated by conventional methods such as preparative chiral chromatography. The compounds may be prepared in racemic form, or each optical isomer may be prepared by stereoselective synthesis or dissolution. For example, the compounds are useful for the formation of salts by selectively active acids such as (-) - di-p-toluoyl-d-tartaric acid and / or (+) - di-p-toluoyl- Such as fraction crystallization and regeneration of the free base after formation of the diastereoisomeric pair. The compound may also be dissolved by chromatographic separation and removal of the chiral auxiliary after formation of the diastereoisomeric ester or amide.

During certain of the methods for preparing the compounds of the present invention it is necessary and / or desirable to protect sensitive or reactive groups on a particular molecule of interest. This is described in Protective Groups in Organic Chemistry, ed. J. F.W. McOmie, Plenum Press, 1973; And T.W. Greene & P.G.M. Can be obtained by conventional protecting groups as described in Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, The protecting group may be removed by conventional sequential steps using methods known in the art.

The compounds of the present invention may be administered to any of the above compositions according to the administration schemes established in the art, wherever certain pharmacological modifications of the activity of muscarinic receptors are required.

The present invention also provides a pharmaceutical composition comprising one or more compounds of the present invention together with a pharmaceutically acceptable diluent or excipient. Preferably, the composition is in unit dosage form for administration by oral gavage (for example, intravenously, intramuscularly or subcutaneously), intrathecal, sublingual or rectal administration, or by inhalation or gas infusion, (Including sustained- or delayed-release formulations), powders, particles, elixirs, tinctures, syrups and emulsions, sterile intestinal fluids or suspensions, aerosol or liquid sprays, drops, ampoules, Injection device or suppository and may be formulated according to the embodiments described in Remington ' s Pharmaceutical Sciences, Gennaro, Ed., Mack Publishing Co., Easton PA, 1990, and in an appropriate manner. Alternatively, the composition is in a sustained-release form suitable for once-weekly or once-monthly dosing; For example, insoluble salts of active compounds such as decanoate salts are suitable for providing diprop preparations for intramuscular injection. The present invention also provides topical formulations suitable for administration, such as in the eye or the skin or mucous membranes.

For example, for oral administration in the form of tablets or capsules, the active drug component may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Suitable binders, lubricants, dispersants, flavoring agents and coloring agents may also be introduced into the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, natural and synthetic gums such as acacia, tragacanth or sodium alginate, carboxymethylcellulose, polyethylene glycol, Do not. Lubricants used in the dosage forms include, but are not limited to, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum and the like.

To prepare a solid composition, such as a tablet, the active ingredient may be admixed with a suitable pharmaceutical excipient as such and other pharmaceutical diluents, such as water, to provide a solid preformulation composition containing a homogeneous mixture of a compound of the invention, To form an acceptable mixture. The term " homogeneous " means that the active ingredient is evenly dispersed throughout the composition such that the composition can be easily degraded into the same effect unit dosage form such as tablets, pills and capsules. The solid preformulation composition may then be reconstituted into unit dosage forms of this kind containing from 0.1 to about 50 mg of the active ingredient of the present invention. The tablets or pills of the present invention are coated or otherwise mixed to provide a dosage form that provides the advantage of prolonged activity. For example, tablets or pills may consist of an inner layer containing the outer layer and the active compound as a coating around the center. The outer coating provides a barrier to inhibit gastrointestinal degradation and is a membrane that allows the inner core to pass through the duodenum while maintaining its original condition or to delay its release. Various materials include mixtures of conventional materials and polymeric acids, such as cellulose, cetyl alcohol and cellulose acetate, and a plurality of polymeric acids, and may be used for coatings or coatings.

The liquid forms in which the composition is incorporated for administration by oral or by injection include not only elixirs but also aqueous solutions, suitably seasoned syrups, aqueous or oil suspensions, and seasoned suspensions with edible oils, such as cottonseed oil, sesame oil, coconut oil, Peanut oil 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 dispersants that may be used include glycerin and the like. For parenteral administration, sterile suspensions and solutions are preferred. Usually isotonic products containing suitable preservatives are used when intravenous administration is desired. The composition may also be formulated as an ophthalmic solution or suspension formulation, such as eye drops, for ocular administration.

As a consequence, the present invention also provides a method for reducing muscarinic receptor activity, particularly m1 receptor activity, which has beneficial effects by administering to a subject in need of such treatment a therapeutically effective amount of a compound of the invention, Or treating a disease or condition. The disease or disorder is due to improper stimulation or activation of muscarinic receptors. By using compounds selective for certain muscarinic receptor subtypes, particularly m1, it is expected that problems with side effects or gastrointestinal effects of known muscarinic drugs such as tachycardia or bradycardia will be substantially avoided.

As used herein, the term " subject " means an animal, preferably a mammal, most preferably a human being, subject of treatment, observation or experiment.

The term " therapeutically effective amount " as used herein refers to a biological or medical response in a tissue, system, animal, or human being sought by a researcher, veterinarian, physician or other clinician, including alleviating the symptoms of the disease being treated ≪ / RTI > of the active compound or pharmaceutical agent.

Preferably, the compounds of formula I exhibit subtype selectivity for the muscarine m1 receptor subtype. Likewise, the compounds exhibit selectivity for the muscarinic m1 receptor subtype compared to other G-caspase-coupled receptors including serotonin, histamine, dopamine or adrenergic receptors. One important point of the selectivity is that the compound is effective in treating or ameliorating many diseases and disorders of the central nervous system without undesirable side effects already found by non-selective compounds.

The ability of the compounds of the present invention to demonstrate muscarinic m1 receptor subtype selectivity is indicative of the ability of the compounds of the invention to be useful in the treatment of attention deficit diseases or neurodegenerative diseases such as cognitive impairment such as Alzheimer's disease or other forms of age- - potentially very useful in treating various diseases and disorders characterized by related symptoms, such as reduced motor function, mood swings, anger, lethargy, insecurity and panic behavior. It is now believed that muscarinic m1 agonists are also used to treat or alleviate eye diseases such as glaucoma, since muscarinic m1 receptors are also involved in controlling intraocular pressure.

The compounds of the present invention may be administered once a day, or the total daily dose administered is divided into two, three or four divided doses per day. The compounds of the present invention may also be administered in the form of a hyperpigmented form by topical application of suitable thickening agents using forms of subcutaneous patches known to those skilled in the art, or via subcutaneous routes. In order to be administered in the form of a subcutaneous delivery system, the dose administration will be more continuous than the short course through the administration schedule.

Dosage regimens employing the compounds of the present invention may be administered by a variety of factors including the type of patient, species, age, weight, sex, and medical condition; The severity of the disease being treated; The route of administration; Renal and hepatic function of the patient; And the particular compound used. A physician or veterinarian of ordinary skill can readily measure and prescribe the effective amount of a drug required to insulate, measure, or stop the progression of the disease or disorder being treated.

The daily dose of the product varies in a wide range from 0.01 to 100 mg per adult per day. For oral administration, the compositions are 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 . Unit doses usually contain from about 0.001 mg to about 50 mg of active ingredient, preferably from about 1 mg to about 10 mg of active ingredient. An effective amount of the drug is orally supplied at a dosage level of about 0.0001 mg / kg to about 25 mg / kg of body weight per day. Preferably, the range is from about 0.001 to about 10 mg / kg of body weight per day and in particular from about 0.001 mg / kg to 1 mg / kg of body weight per day. The compounds are administered in a schedule of 1 to 4 times per day.

The compounds according to the invention are used at moderate doses as defined by the pathway test to obtain optimal pharmaceutical effects on muscarinic receptors, particularly muscarinic m1 receptor subtypes, while minimizing potential toxicity or other undesirable effects. And co-administration or sequential administration of other agents that improve the effectiveness of the compounds is preferred in some cases.

The pharmaceutical properties and selectivity of the compounds of the present invention for a particular muscarinic receptor subtype can be assessed using a variety of different assays using recombinant receptor subtypes, preferably with conventional secondary messenger or binding assays, ≪ / RTI > Certain prior art functional assay systems are described in U.S. Pat. No. 5,101,505, which describes a method for screening for bioactive compounds using the ability of cells infected by receptor DNA encoding other muscarinic subtypes, which can be amplified in the presence of a ligand of the receptor Is a receptor selection and amplification method as disclosed in US 5,707,798. Cell amplification is also detected by the cell as an increased level of the expressed marker.

The present invention is described in further detail in the following examples which are not to be construed as limiting the scope of the invention as claimed, in any way.

Example 1

4-n-butyl-1- [4- (2-methylphenyl) -4-oxo-1-butylpiperidine (5)

1-Benzyl-4-n-butylidenepiperidine (2). Sodium hydride (1.61 g, 67 mmol) and DMSO (40 mL) are charged to a 500 mL three-necked flask equipped with a stirrer. The resulting suspension is heated to < RTI ID = 0.0 > 90 C < / RTI > for 30 minutes until evolution of hydrogen ceases. The suspension is cooled in an ice-bath for 20 min and a slurry of butyltriphenylphosphonium bromide (26.6 g, 67 mmol) in DMSO (70 ml) is added. The red mixture is stirred at room temperature for 15 minutes. 1-Benzyl-4-piperidone (1) (14.0 g, 74 mmol) is slowly added over 30 minutes and the mixture is stirred at room temperature overnight. H ₂ O (200 mL) is added to the reaction mixture and extracted with heptane (4 × 100 mL) and ethyl acetate (2 × 100 mL). The combined organic phases are dried and evaporated to dryness, yielding 38.1 g of a yellow oil. The oil was distilled to give 14.9 g (88%) of (2), bp 101-105 ° C (0.1 mmHg). _{^{1 H NMR (CDCl 3) δ0.90-0.95}} (t, 3H), 1.25-1.41 (m, 2H), 1.90-2.20 (m, 2H), 2.18-2.30 (m, 4H), 2.40-2.45 (m , 4H), 2.50 (s, 2H), 5.17 (t, 1H), 7.20-7.42 (m, 5H).

4-n-butylpiperidine (3). To a 500 ml flask equipped with a stirrer was added a slurry of 10% palladium and (2) (13.2 g, 58 mmol) in charcoal (1.2 g) in ethanol (70 ml) and concentrated hydrochloric acid (1.5 ml) . The reaction flask is removed and hydrogen is added through the reaction flask. A total of 2.5 dm 3 of hydrogen is consumed. The reaction mixture was filtered, evaporated and the residue was dissolved in H ₂ O (40 mL) and NaOH (20 mL, 2M) and extracted with ethyl acetate (3 × 100 mL). The combined organic phases are washed with brine (30 ml) and evaporated to dryness to yield 7.1 g of crude product (3). The crude product is treated CC (eluent: heptane: EtOAc (4:. 1) ], pure (3) (2.7g, to yield ^{33%) 1 H NMR (CDCl} 3) δ0.85 (t, 3H) 2H), 1.01- 1.38 (m, 9H), 1.65 (dd, 2H), 2.38 (s, 1H), 2.55 (dt, 2H), 3.04 (dt, 2H).

4- (4-n-butylpiperidin-1-yl) butanenitrile (4). (2.3 g, 16.4 mmol), 4-bromobutyronitrile (2.4 g, 16.4 mmol) and potassium carbonate powder (2.5 g, 18 mmol) in acetonitrile (20 ml) were added to a 100 ml flask equipped with a magnetic stirrer, . The reaction mixture is stirred at room temperature for 5 hours and H ₂ O (15 ml) is added. The mixture is extracted with ethyl acetate (3 x 30 ml) and the combined organic phases are evaporated to dryness to give 3.9 g of crude product (4). The crude product is treated CC (eluent: heptane: EtOAc (1:. 1) ], pure (4) (2.3g, to give a ^{87%) 1 H NMR (CDCl} 3) δ0.82 (t, 3H ), 1.19-1.37 (m, 9H), 1.64-1.75 (d, 2H), 1.84-2.01 (m, 4H), 2.39-2.54 (m, 4H), 2.89-2.97 (d, 2H).

4-n-Butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine (5). A 25 ml oven-dried flask is filled with Mg turning (125 mg, 5.2 mmol) activated by using a heat-gun. Under an inert atmosphere, a suspension of 2-iodoanisole (1.13 g, 5.2 mmol) in Et ₂ O (4 ml) is added and the reaction mixture is allowed to stand at room temperature for 1 h. Compound (4) (720 mg, 3.4 mmol) dissolved in Et ₂ O (4 ml) is added and the mixture is refluxed overnight. THF (15 mL) and sulfuric acid (4 mL, 2M) are added and the reaction mixture is stirred for 4 hours and NaOH (6 mL, 2M) is added. The reaction mixture is extracted with ethyl acetate (3 x 50 ml) and the combined organic phases are evaporated to dryness, yielding 1.2 g of crude product (5). The crude product is CC-treated (eluent: CH ₂ Cl ₂ : CH ₃ OH (99: 1)] to give pure (5) (0.42 g, 26%) ¹ H NMR (CDCl ₃ ) δ0. 4H), 2.53 (t, 2H), 3.02-3.17 (m, 4H), 1.63-1.30 (m, 3.89 (s, 3H), 6.97-7.01 (m, 2H), 7.44 (t, 1H), 7.65 (d, 1H).

Example 2

3-hydroxymethyl- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine (7)

4- (3-Hydroxymethyl-piperidin-l-yl) -butyronitrile (6). To a 25 mL oven-dried flask was added piperidine-3-yl-methanol (1.12 g, 10 mmol) in acetonitrile (10 mL) and potassium carbonate (1.38 g, 10 mmol) and 4- bromobutyronitrile (0.90 ml, 9 mmol). The reaction mixture is stirred at room temperature for 12 hours. The mixture is filtered and evaporated to dryness. H ₂ O (20 ml) was added and extracted with ethyl acetate (3 x 20 ml) and the combined organic phases were dried (MgSO ₄ ) and evaporated to dryness to afford further purification in the synthesis of compound (7) 1.50 g of crude product 6, which can be used without further purification, is obtained.

3-Hydroxymethyl- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine (7). To an oven-dried 50 ml flask was added Mg turning (780 mg, 32 mmol) activated using a heat-gun under vacuum and anhydrous THF (7 ml) was added. Under an inert atmosphere, a suspension of 2-iodotoluene (5.3 g, 24 mmol) in THF (10 ml) is added and the reaction mixture is refluxed for 4 hours. A suspension of compound 6 (1.50 g, 8 mmol) in THF (5 ml) was added via an injector and CuBr (23 mg, 0.16 mmol, 2 mol%) was added and the reaction mixture was stirred overnight at room temperature do. The reaction mixture is quenched by the addition of H ₂ SO ₄ (20 mL, 2M) and stirred at room temperature for 2 h before NaOH (8 mL, 2M) is added. And sulfuric acid (4 mL, 2M) are added and the reaction mixture is stirred for 4 hours and NaOH (6 mL, 2M) is added. THF (15 ml) is added, extracted with CH ₂ Cl ₂ (3 × 20 ml), the organic phase is dried (MgSO ₄ ) and evaporated to dryness, yielding 0.41 g of crude product 7. The crude product was preliminarily HPLC CC treated [eluent: Buffer A: 0.1% TFA; Buffer _{B: 80% CH 3 CN +} 0.1% TFA], a pure sample is generated analytically of compound (7). LC-MS [M + H] < ⁺ > 275 (cald.

Example 3

2-propyl- [4- (2-methylphenyl) -4-oxo-1-butylpiperidine (9)

4- (2-Propyl-piperidin-l-yl) -butyronitrile (8). (550 mg, 4.3 mmol), 4-bromobutyronitrile (430 mg, 3.0 mmol) and potassium carbonate (550 mg, 4.0 mmol) in acetonitrile (5 ml) Stir at temperature and add saturated brine (25 mL). The reaction mixture is extracted with ethyl acetate (3 x 25 mL), the combined organic phases are dried (MgSO ₄ ) and evaporated to dryness to yield the crude product 8. The crude product is CC-treated to give pure (8) (0.48 g, 83%) [eluent: CH ₂ Cl ₂ : MeOH (99: 1)]; LC-MS [M + H] < ⁺ > 194 (Cald. 194.2).

2-Propyl- [4- (2-methylphenyl) -4-oxo-1-butyl] piperidine (9). To an oven-dried 10 ml flask add Mg turning (97 mg, 4.1 mmol) activated under vacuum using a heat-gun. Under an inert atmosphere, a suspension of 2-iodotoluene (380 L, 2.8 mmol) in Et ₂ O (3 mL) is added and the reaction mixture is refluxed for 1 hour. A mixture of compound (8) (0.43 g, 2.2 mmol) in CH ₂ Cl ₂ (3 mL) was added via an injector and the reaction mixture was stirred at room temperature overnight. The reaction mixture is quenched by the addition of H ₂ SO ₄ (10 mL, 2M), stirred at room temperature for 12 h, and then NaOH (10 mL, 2M) is added. The combined organic phases were washed with brine (10 mL) and NaOH (10 mL, 2M), dried (MgSO ₄ ), dried (10 mL) Evaporation on the road yields 0.43 g of crude product 9. The crude product was subjected to preparative HPLC [eluent: Buffer A: 0.1% TFA; Buffer _{B: 80% CH 3 CN +} 0.1% TFA], a pure sample of compound 9 is generated analytically. LC-MS [M + H] < ⁺ > 287 (Cald 287.2).

Example 4

1- [4- (2-methylphenyl) -4-oxo-1-butylpiperidine (11)

To an oven-dried 10 ml flask add Mg turning (97 mg, 4.1 mmol) activated under vacuum using a heat-gun. Under an inert atmosphere, a suspension of 2-iodo-toluene (380 L, 3.0 mmol) in Et ₂ O (3 mL) is added and the reaction mixture is refluxed for 1 hour. (Dahlbom et al., Acta. Chem. Scand. 1951, 5, 690-697) (0.305mg, 2.0mmol) in CH ₂ Cl ₂ (3ml) was added to a solution of 4-piperidin- Is added via an injector, and the reaction mixture is stirred at room temperature overnight. The reaction mixture is cooled by adding H ₂ SO ₄ (10 mL, 2M), stirred at room temperature for 12 h, and then NaOH (12 mL, 2M) is added. The combined organic phases were washed with brine (10 mL) and NaOH (10 mL, 2M), dried (MgSO ₄ ), dried (10 mL) Evaporation on the road gives 0.21 g of crude product 11. The crude product was subjected to preparative HPLC [eluent: Buffer A: 0.1% TFA; Buffer _{B: 80% CH 3 CN +} 0.1% TFA], a pure sample of compound 11 is generated analytically. LC-MS [M + H] < ⁺ > 245 (cald.

Example 5

Methyl-1- [4- (4-bromophenyl) -4-oxo-1-butylpiperidine (12)

4-methylpiperidine (719,, 6 mmol) and dioxane (5 ml) were added to a 10 ml dried flask and potassium carbonate (0.30 g, 2.18 mmol), potassium iodide Mo-4-chlorobutyrophenone (785 mg, 2.76 mmol) is added. The reaction mixture is allowed to stand at 110 ℃ for 12 hours, then diluted with H ₂ O (10㎖). The reaction mixture is extracted with Et ₂ O (3 × 15 ml) and the combined organic phases are dried (MgSO ₄ ) and evaporated to dryness, yielding 0.50 g of crude product 12. The crude product was subjected to preparative HPLC [eluent: Buffer A: 0.1% TFA; Buffer _{B: 80% CH 3 CN +} 0.1% TFA], a pure sample is generated analytically of compound (12); LC-MS [M + H] < ⁺ > 322 (Cald. 323.1).

Example 6

1- [4- (2-methylphenyl) -4-oxo-1-butylpyrrolidine (13)

The oven-dried 10 mL flask was charged with Mg turning (30 mg, 1.2 mmol) activated using a heat-gun under vacuum. Under an inert atmosphere, a solution of 2-iodotoluene (0.22 g, 1.0 mmol) in Et ₂ O (2 mL) is added and the reaction mixture is refluxed for 1 hour. A mixture of 4-pyrrolidin-1-yl-butyronitrile (Burkhalter et al., J. Org. Chem. 1961, 26, 4070-4076) (0.14 g, 1.0 mmol) in CH ₂ Cl ₂ Is added via an injector, and the reaction mixture is stirred at room temperature overnight. The reaction mixture is cooled by the addition of H ₂ SO ₄ (10 mL, 2M), stirred at room temperature for 2 h, and then NaOH (10 mL, 2M) is added. THF (15 mL) is added, extracted with ethyl acetate (3 x 20 mL), the organic phase is dried (MgSO ₄ ) and evaporated to dryness, yielding 0.12 g of crude product 13. The crude product was subjected to preparative HPLC [eluent: Buffer A: 0.1% TFA; Buffer _{B: 80% CH 3 CN +} 0.1% TFA], a pure sample of compound 13 is generated analytically. LC-MS [M + H] < ⁺ > 231 (cald.

Example 7

Methyl-1- [4- (2-methylphenyl) -4-oxo-1-butylpiperazine (15)

4- (4-Methyl-piperazin-1-yl) -butyronitrile (14). To a 25 mL flask was added suspended 1-methyl-piperazine (0.52 g, 5.1 mmol), 4-bromobutyronitrile (0.78 g, 5.3 mmol) and potassium carbonate (0.71 g, 5.3 mmol) in acetonitrile mmol). The reaction mixture is stirred at room temperature for 4 h, H ₂ O (20 ml) is added and extracted with ethyl acetate (3 x 25 ml). The combined organic phases are washed with brine (25㎖), dried (MgSO _4), and generates a crude product (14) 0.72g, which can be used without further purification in the synthesis of the road evaporated to dryness, Compound (15) .

4-methyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine (15). To a 10 ml oven-dried flask is added Mg turning (116 mg, 4.0 mmol) which is activated using a heat-gun under vacuum. Under an inert atmosphere, a mixture of 2-iodotoluene (0.65 g, 3.0 mmol) in Et ₂ O (3 ml) is added and the reaction mixture is refluxed for 1 h. A solution of compound (14) (0.33 g, 2.0 mmol) in CH ₂ Cl ₂ (3 mL) is added via an injector and the reaction mixture is stirred at room temperature overnight. The reaction mixture is quenched by the addition of H ₂ SO ₄ (60 mL, 2M), stirred at room temperature for 2 h, and then NaOH (8 mL, 2M) is added. Add THF (15 mL) and extract with CH ₂ Cl ₂ (3 x 20 mL). The organic phase is dried (MgSO ₄ ) and evaporated to dryness, yielding 0.26 g of crude product 15. The crude product was subjected to preparative HPLC [eluent: Buffer A: 0.1% TFA; Buffer _{B: 80% CH 3 CN +} 0.1% TFA], a pure sample is generated analytically of compound (15). LC-MS [M + H] < ⁺ > 260 (cald. 260.4).

Example 8

4-n-Butyl-1- [4- (2-methylphenyl) -4-oxo-

4- (4-Butyl-piperazin-1-yl) -butyronitrile (16). To a 25 mL flask was added 1-butyl-piperazine (712 mg, 5.0 mmol), 4-bromobutyronitrile (779 mg, 5.3 mmol) suspended in acetonitrile (5 mL) and potassium carbonate mmol). The reaction mixture is stirred at room temperature for 12 hours, H ₂ O (20 ml) is added and extracted with ethyl acetate (3 × 25 ml). The combined organic phases are washed with brine (25 mL), dried (MgSO ₄ ) and evaporated to dryness to yield 0.89 g of crude product 16, which can be used without further purification in the synthesis of compound 17.

4-n-Butyl-1- [4- (2-methylphenyl) -4-oxo-1-butyl] piperazine (17). Into a 10 ml oven-dried flask is added Mg turning (100 mg, 4.0 mmol) activated using a heat-gun under vacuum. Under an inert atmosphere, a suspension of 2-iodotoluene (0.66 g, 3.0 mmol) in Et ₂ O (3 mL) is added and the reaction mixture is refluxed for 1 h. A suspension of compound (16) (0.43 g, 2.0 mmol) in CH ₂ Cl ₂ (3 mL) was added via an injector and the reaction mixture was stirred at room temperature overnight. The reaction mixture is quenched by the addition of H ₂ SO ₄ (6 mL, 2M) and stirred at room temperature for 2 h before NaOH (8 mL, 2M) is added. THF (15 ml) is added, extracted with CH ₂ Cl ₂ (3 × 20 ml), the organic phase is dried (MgSO ₄ ) and evaporated to dryness to give 0.50 g of crude product 17. The crude product was subjected to preparative HPLC [eluent: Buffer A: 0.1% TFA; Buffer _{B: 80% CH 3 CN +} 0.1% TFA], a pure sample is generated analytically of compound (17). LC-MS [M + H] < ⁺ > 302 (Cald.

Example 9

4-n-Butyl-1- [4- (2-ethoxyphenyl) -4-oxo-1-butylpiperidine (18)

To an oven-dried 10 ml flask was added Mg turning (94 mg, 3.8 mmol) which was activated using a heat-gun under vacuum. Under an inert atmosphere, a suspension of 1-ethoxy-2-iodobenzene (0.71 g, 2.9 mmol) in Et ₂ O (3 mL) is added and the reaction mixture is refluxed for 3 hours. Compound (4) (0.40 g, 1.9 mmol) dissolved in CH ₂ Cl ₂ (3 mL) is added and the mixture is stirred at 40 ° C. for a further 3 hours. The reaction mixture is quenched by the addition of H ₂ SO ₄ (10 mL, 2M), stirred at room temperature overnight, and NaOH (20 mL, 2M) is added until basic conditions occur. The combined organic phases were washed with brine (10 mL) and NaOH (10 mL, 2M) and the combined organic phases were dried (MgSO ₄ ) Evaporation yielded 0.60 g of crude product 18. The crude product was CC-treated to give [Eluent: Tol: EtOAc (1: 1)], pure compound 18 (0.32 g, 34%); LC-MS [M + H] < ⁺ > 331 (Cald 331.5).

Example 10

Butyl-1- [4- (2,3-dimethylphenyl) -4-oxo-1-butyl] piperidine (19)

To an oven-dried 10 ml flask add Mg turning (94 mg, 3.8 mmol) which is activated by using a heat-gun under vacuum. Under an inert atmosphere, also in 1-iodo Et ₂ O (5㎖) a suspension of 2,3-dimethyl-benzene (0.69g, 3.0mmol) is added under automatic reflux, the reaction mixture is refluxed for 4 hours. A suspension of compound 4 (0.41 g, 2.0 mmol) in CH ₂ Cl ₂ (2 mL) is added to the reaction mixture and left at room temperature overnight. The reaction mixture is quenched by the addition of H ₂ SO ₄ (7 mL, 2M), stirred at room temperature for 3 hours, and then NaOH (20 mL, 2M) is added until basic conditions occur. The combined organic phases were washed with brine (10 mL) and NaOH (10 mL, 2M) and the organic phase was dried (MgSO ₄ ) and evaporated to dryness , 0.69 g of the crude product 19 is produced. The crude product is CC-treated to give pure [EA: CH ₂ Cl ₂ : MeOH (99: 1)], pure 19 (0.40 g, 64%); LC-MS [M + H] < ⁺ > 315 (Cald 315.5).

Example 11

4-n-butyl-1- [4- (2,4-dimethylphenyl) -4-oxo-1-butyl] piperidine (20)

The oven-dried 10 mL flask is filled with Mg turning (95 mg, 3.9 mmol) activated using a heat-gun under vacuum. Under an inert atmosphere, a suspension of 1-iodo-2,4-dimethylbenzene (0.69 g, 2.9 mmol) in Et ₂ O (4.5 ml) is added under spontaneous reflux and the reaction mixture is refluxed for 3 hours. Compound (4) (0.41 g, 2.0 mmol) dissolved in CH ₂ Cl ₂ (2 mL) is added to the reaction mixture under an inert atmosphere and stirred at room temperature overnight. The reaction mixture is quenched by the addition of H ₂ SO ₄ (8 mL, 2M), stirred at room temperature for 4 hours and then basified by addition of NaOH (20 mL, 2M). The combined organic phases were washed with brine (10 mL) and NaOH (10 mL, 2M) and the organic phase was dried (MgSO ₄ ) And evaporated to dryness, yielding 0.61 g of crude product 20. The crude product is CC-treated to give [eluent: CH ₂ Cl ₂ : MeOH (99: 1)], pure compound 20 (0.21 g, 35%); LC-MS [M + H] < ⁺ > 315 (Cald 315.5).

Example 12

Butyl-1- [4- (2-methoxyphenyl) -4-oxo-1-butyl] piperidine (21)

The oven-dried 10 mL flask is charged with Mg turning (0.12 g, 4.9 mmol) activated using a heat-gun under vacuum. Under an inert atmosphere, a suspension of 1-bromo-2-ethylbenzene (0.66 g, 3.6 mmol) in Et ₂ O (2 mL) is added and the reaction mixture is refluxed for 2 hours. A suspension of compound (4) (0.50 g, 2.4 mmol) in CH ₂ Cl ₂ (2 mL) was added via an injector and the reaction mixture was stirred overnight at room temperature. The reaction mixture is quenched by the addition of H ₂ SO ₄ (14 mL, 2M), stirred at room temperature for 2 hours, and then NaOH (20 mL, 2M) is added. The combined organic phases are washed with brine (10 mL) and NaOH (10 mL, 2M), the organic phase is dried (MgSO ₄ ) And evaporated to dryness, yielding 0.75 g of crude product 21. The crude product was CC-treated to give pure [Eta] l (CH ₂ Cl ₂ : MeOH (99: 1)], compound 21 (0.68 g, 90%); LC-MS [M + H] < ⁺ > 315 (Cald 315.5).

Example 13

Butyl-1- [4- (2,4-dimethylphenyl) -4-oxo-1-butyl] piperidine (22)

The oven-dried 10 mL flask was charged with Mg turning (88 mg, 3.6 mmol) activated using a heat-gun under vacuum. Under an inert atmosphere, a suspension of 1-iodo-2-methoxymethylbenzene in Et ₂ O (4 mL) (0.67 g, 2.7 mmol) is added and the reaction mixture is refluxed for 1 h. A suspension of compound (8) (0.38 g, 1.8 mmol) in CH ₂ Cl ₂ (4 mL) was added via an injector and the reaction mixture was stirred at room temperature overnight. The reaction mixture is quenched by the addition of H ₂ SO ₄ (10 mL, 2M), stirred at room temperature for 2 hours, and then NaOH (10 mL, 2M) is added. The combined organic phases are washed with brine (10 mL) and NaOH (10 mL, 2M), the organic phase is dried (MgSO ₄ ) And evaporated to dryness, yielding 0.51 g of crude product 22. The crude product is CC-treated to give pure [22] (0.14 g, 23%) [eluent: CH ₂ Cl ₂ : MeOH (99: 1)]; LC-MS [M + H] < ⁺ > 331 (Cald 331.5).

Example 14

Butyl-1- [4- (2-pyridinyl) -4-oxo-1-butyl] piperidine (24)

4- (4-Butyl-piperidin-l-yl) -butyric acid methyl ester (23). To the reaction flask 25㎖ CH ₃ CN (10㎖) within the suspension of 4-bromo-butyric acid methyl ester (2.04g, 11.2mmol), compound (3) (1.51g, 10.8mmol) and potassium carbonate (1.63g, 11.8 mmol). The reaction mixture is stirred overnight at room temperature, filtered and evaporated to dryness. Add H ₂ O (50 mL) and extract with ethyl acetate (3 x 100 mL). The combined organic phases were dried (MgSO _4), evaporated and dried road, a 2.84g of crude product (23) is generated. The crude product is CC-treated (eluent: CH ₂ Cl ₂ : MeOH (99: 1)) to give pure compound 23 (1.93 g, 75%). LC-MS [M + H] < ⁺ > 241 (Cald. 241.2).

4-n-Butyl-1- [4- (2-pyridinyl) -4-oxo-1-butyl] piperidine (24). To a 25 mL dry reaction flask is added dissolved 2-bromopyridine (200 mg, 1.3 mmol) in CH ₂ Cl ₂ (3 mL) and the temperature is adjusted to -78 ° C. After stirring for 20 minutes, n-BuLi (0.84 mL, 1.4 mmol) is added under an inert atmosphere. After an additional 30 minutes, a solution of compound (23) dissolved in CH ₂ Cl ₂ (2 mL) is added. The reaction mixture is warmed to room temperature overnight before cooling with H ₂ SO ₄ (5 mL, 1M). The reaction mixture is extracted with ethyl acetate (6 × 25㎖), the combined organic phases dried (MgSO _4), evaporated to dryness road, a 0.31g of crude product (24) is generated. The crude product was CC-treated to give pure [242] (75 mg, 12%) [eluent: CH ₂ Cl ₂ : MeOH (10: 1)]; LC-MS [M + H] < ⁺ > 288 (Cald 288.2).

Example 15

Butyl-1- [4- (2-hydroxyphenyl) -4-oxo-1-butyl] piperidine (27)

1-Benzyloxy-2-iodo-benzene (25). 2-Iodophenol (1.03 g, 4.7 mmol) and potassium carbonate (0.71 g, 5.2 mmol) are dissolved in dry acetone (10 ml) in a 25 ml oven-dried flask. The mixture is stirred for 15 minutes, benzyl bromide (0.61 mL, 5.2 mmol) is added and left at room temperature overnight. H ₂ O (50 ml) was added, extracted with ethyl acetate (3 × 50 ml) and the combined organic phases were dried (MgSO ₄ ) and evaporated to dryness to yield 1.7 g of crude product 25 . The crude product is CC-treated (eluent: heptane: EtOAc (9: 1)) to give pure compound 25 (1.2 g, 81%). LC-MS [M + H] < ⁺ > 310 (cald. 310.0).

4-n-Butyl-1- [4- (2-benzyloxyphenyl) -4-oxo-1-butyl] piperidine (26). Activated Mg turning (123 mg, 5.1 mmol) is added to a 25 ml oven-dried flask using a heat-gun under vacuum. A solution of 1-benzyloxy-2-iodo-benzene (25) (1.18 g, 3.8 mmol) in Et ₂ O (10 mL) is added under an inert atmosphere and the reaction mixture is refluxed for 3.5 hours. A solution of the dissolved 4- (4-n-butylpiperidin-l-yl) -butanenitrile 4 (0.53 g, 2.5 mmol) in CH ₂ Cl ₂ (3 ml) Lt; 0 > C overnight. The reaction mixture is cooled by the addition of H ₂ SO ₄ (10 mL, 2M), stirred for 1 hour, and NaOH (20 mL, 2M) is added until basic conditions are reached. The combined organic phases were washed with brine (10 mL) and NaOH (10 mL, 2M) and the combined organic phases were dried (MgSO ₄ ), dried Evaporation on the road yields 1.28 g of crude product 26. The crude product was CC-treated to give [Eluent: Tol: EtOAc (1: 1)], pure compound 26 (0.51 g, 51%); LC-MS [M + H] < ⁺ > 393 (Cald 393.7).

4-n-butyl-1- [4- (2-hydroxyphenyl) -4-oxo-1-butyl] piperidine (27). To a 25 ml reaction flask was added 4-n-butyl-1- [4- (2-benzyloxyphenyl) -4-oxo-1-butyl] piperazine dissolved in dry EtOH (10 ml) A solution of feridine 26 (49 mg, 1.2 mmol) is added and palladium (40 mg) is added onto charcoal. The reaction flask is then filled with H ₂ using a mechanical type technique and stirred at room temperature overnight under H ₂ atmosphere. The reaction mixture is basified by the addition of NaOH (2 mL, 2.0 M) and filtered through celite. The aqueous phase was extracted with ethyl acetate (3 x 50 mL) and the combined organic phases were washed with brine (10 mL) and NaOH (10 mL, 2M), dried (MgSO ₄ ) and evaporated to dryness, 0.42 g of the product 27 is produced. The crude product was CC-treated to give [Eluent: CH ₂ Cl ₂ : MeOH (99: 1)], pure compound 27 (0.21 g, 58%); LC-MS [M + H] < ⁺ > 303 (cald.

Example 16

Method of screening test compounds in assays using muscarinic receptor subtypes m1, m2, m3, m4 and m5

Gene infection of cells with muscarinic receptor DNAs (common method)

NIH 3T3 cells (available as ATCC CRL 1658 from American Type Culture Collections) were added at a concentration of 4.5 g / l glucose, 4 mM glutamine, 50 units / ml penicillin, 50 units / ml streptomycin (Advanced Biotechnologies, Inc., Gadsburg, MD). (5% CO ₂ ) incubator in a humidified atmosphere (5% CO ₂ ) incubator in a controlled Dulbecco's Eagle medium supplemented with 10% fetal calf serum (Sigma, St. Louis, . The cells are treated with trypsin-EDTA, spun and 2 x 10 ⁶ per 15 cm dish in 20 ml DMEM containing 10% calf serum.

The m1-m5 muscarinic receptor subtype is cloned in the same manner as described in Bonner et al., Science 237, 1987, page 527, and Bonner et al., Neuron 1, 1988, m2 and m4 receptors, the cells are co-transfected together with DNA encoding a chimera between the Gq protein and the five carboxy-terminal amino acids of the Gi protein (Gq-i5 constructs are described in Conklin et al., Nature 363, 1993, Lt; / RTI >

The cells are genetically infected using a Superfect gene infection reagent (Valencia Qiagen, Calif.) According to the manufacturer's instructions. Chimeric Gq-i5 DNA for receptor DNA, β-gal DNA (pSI-β-galactosidase from Promega, Madison, Wis.), M2 and m4 receptor subtypes, and salmon sperm DNA Sigma, Lewis) is added to a total of 20 ug of DNA per plate. Before adding to the plate, 60 μl of the supernatant is added to the DNA, and the pipette is shaken up and down several times to thoroughly mix. The mixture is incubated at room temperature for 10-15 minutes. The medium is aspirated and 12 ml of fresh DMEM containing 10% calf serum and 50 units / ml penicillin / streptomycin are added to the plate. The DNA-Perfect solution is mixed once more with a pipette, and the vial is added to the plate so that the DNA mixture is uniformly distributed on the surface. The cells are incubated overnight at 37 ° C and 5% CO ₂ .

After incubation, the medium is aspirated and the plate is washed once with 15 ml of Hank's buffered saline solution. The plate is agitated to thoroughly wash. 20 ml of fresh DMEM supplemented with 10% calf serum and 50 units / ml penicillin / streptomycin are added to the plate. The cells are cultured for 24-48 hours until the plates are 100% fused.

Analysis of NIH 3T3 cells gene-infected with muscarinic receptor subtypes (general method)

DMEM containing 2% Cyto-SF3 is heated at 37 占 폚 in sterile water. A sterile working stock solution of the test compound to be analyzed is prepared to dilute the compound in DMEM to a test 8x final concentration. The compound (carbachol) included in the assay as a positive control is diluted to a final concentration of 8X in DMEM. 50 [mu] l of DMEM containing 2% Cyto-SF3 is added to a 96-well microtiter plate under sterile conditions. Then, 16 μl of the compound solution is added to the upper well of the plate, and the solution is diluted by taking 16 μl of the compound solution in the upper well and pipetting into the wells of the next row. Although the method is repeated in the heat of each successive well, 50 mu l of the medium itself is used as the baseline control well (containing the medium and cells, but not the test compound) and plate control wells (wells containing medium, But not test compounds and cells). The plate is placed in an incubator at 37 ° C to equilibrate the temperature and pH.

When the cell culture is 100% fused, the medium is aspirated and each plate is washed with 15 ml of Hank's buffered saline (HBS). The cells are left in the incubator for about 10-15 minutes until the HBS turns slightly yellow. The HBS is withdrawn, 1 ml of trypsin is added to each plate, and the plate is whipped completely over. The edges of the plate are tapped several times to break down the cells. After the cells are removed from the surface, 8 ml of DMEM containing 10% calf serum and 50 units / ml penicillin and 50 units / ml streptomycin are added to inhibit trypsin. The plate is washed with a medium, and the cells are pipetted into a tube. The cells are centrifuged at 1000 rpm for 5 to 10 minutes on an IEC Centra CL2 centrifuge (Sorvall). The medium is then carefully aspirated to remove cells. The cell pellet is suspended in 1600 [mu] l DMEM containing 10% calf serum and 50 units / ml penicillin and 50 units / ml streptomycin and 20 ml DMEM supplemented with 2% Cyto-SF3 are added. An aliquot of 50 [mu] l of the cell suspension is added to the wells of the 96-well microtiter plate prepared above (except for plate control wells). And the plate is incubated at 37 ℃ and 5% CO ₂ for 4 days.

After incubation, the medium is removed by converting the microtiter plate and gently shaking it, which is then blotted on a sorbent paper. 100 μl of chromogen material (3.5 mM o-nitrophenyl-β-D-galactopyranoside in phosphate buffered saline, 0.5% nonidet NP-40) was added to each well and the plate was incubated at 405 nm with an appropriate absorbance Lt; RTI ID = 0.0 > 30 C. < / RTI > Absorbance of the baseline and plate control wells is subtracted from all values.

result

Using the general method described above, NIH 3T3 cells are genetically infected with DNAs encoding the m1, m3 and m5 receptor subtypes. A compound library containing approximately 35,000 small organic compounds (one per well) is screened against the receptor by the method described above. Figure 1 shows data from one 96-well plate from a screen. In this plate, two compounds are activated at one or more gene-infected receptors. On the whole screen, four related compounds were found to exhibit activity. To determine that the receptor is activated on the screen, the compound is tested as described above for the gene-infection of each receptor with a separate cell culture. Only Compound A activates the m1 receptor subtype and is a partial potential agent that induces a lower maximum response than the control compound, carbachol.

In a further experiment, four compounds were found to selectively activate the m1 receptor which does not exhibit significant activity at m2, m3, m4 or m5 muscarinic receptors. Compound A, which is the most active compound, is not an antagonist of the carbachol-induced response of the five muscarinic receptor subtypes.

Compound A is additionally tested for agonist activity against several other receptors in the alpha -adrenaline receptor subtypes 1D, 1B, 1A, 2A, 2B and 2C, histamine H1 and serotonin 5-HT1A and 5-HT2A subtypes. The compounds do not show significant activity in the assay. In antagonist testing, Compound A did not inhibit the response of alpha -adrenaline receptor subtype 2A, 2B or 2C, or the serotonin receptor subtype 5-HT1A or 5-HT2A. As illustrated in FIG. 2, the reaction induced by Compound A is blocked by the muscarinic antagonist atropine, as in the reaction induced by muscarinic antagonistic carbachol.

Example 17

R-SAT Analysis

R-SAT analysis (incorporated herein by reference, see U.S. Patent No. 5,707,798) is carried out by exposing cells transfected with the m1, m3 or m5 receptor to seven compounds at a concentration of 1.5 μM. The cellular response is expressed as a ratio to the maximal response of the cells (defined as the response to 10 μM carbachol). The results are shown in the following table.

compound Receptor and Concentration m1 1.5 [mu] M m3 1.5 [mu] M m5 1.5 μM A (Example 1) 107 +/- 9 7 +/- 8 3 +/- 8 B (Example 4) 76 +/- 11 7 +/- 9 -6 +/- 10 C (Example 15) 91 +/- 9 4 +/- 9 0 +/- 12 D (Example 10) 72 +/- 9 13 +/- 7 2 +/- 15 E (Example 11) 42 +/- 13 9 +/- 3 -3 +/- 2 F (Example 12) 65 +/- 9 9 +/- 7 5 +/- 11 G ^* 66 +/- 19 16 +/- 12 7 +/- 11

* 4-n-Butyl-1- [4-phenyl-4-oxo-1-butyl] piperidine

As described above, the compound is an selective agonist of the m1 receptor.

The invention described hereinabove and claimed herein is not to be limited by the specific embodiments described above, as such embodiments are illustrative of some aspects of the invention. Certain embodiments are intended to be included within the scope of the present invention. Indeed, various modifications of the invention, as well as those described and disclosed herein, will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.

A variety of literature is cited here, and his specializations are incorporated by reference.

Claims (99)

Or a pharmaceutically acceptable salt, ester or prodrug thereof. (I) Wherein X ₁ , X ₂ , X ₃ , X ₄ and X ₅ are selected from C, N and O; k is 0 or 1; t is 0, 1 or 2; R ₁ is straight or branched C _1-8 alkyl, C _2-8 alkenyl, C _1-8 alkynyl, C _1-8 alkylidene, C _1-8 alkoxy, C _1-8 heteroalkyl, C _{1- 8} aminoalkyl, C _1-8 haloalkyl, C _1-8 alkoxycarbonyl, C _1-8 hydroxy alkoxy, C _1-8 hydroxyalkyl, -SH, C _1-8 alkylthio, -O-CH ₂ -C (O) -C _5-6 aryl substituted with C _5-6 aryl, C _1-3 alkyl or halo; C _5-6 aryl or C _5-6 cycloalkyl optionally containing one or more heteroatoms selected from N, S and O; _{-C (O) NR 3 R 4} , -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -CR 3 R 4, -OC (O) R 3, - (O) (CH 2) _s NR ₃ R ₄ or _{- (CH 2) s NR 3} R 4 is (wherein, R _3, R ₄ and R ₅ are the same or different and each H, C _1-6 alkyl, from N, O or S selected and, optionally, halo, or C _5-6 aryl optionally substituted with one or more heteroatom containing the character C _1-6 alkyl is selected from C _3-6 cycloalkyl; or R ₃ and R ₄ taken together with the N atom If present, forms a cyclic ring structure comprising 5-6 atoms selected from C, N, S and O; s is an integer from 0 to 8; A is N, C, including the at least one heteroatom selected from S and O optionally _5-12 aryl or C _5-7 cycloalkyl; R ₂ is H, amino, hydroxyl, halo, or straight or branched C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 heteroalkyl, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _1-6 alkoxycarbonyl, C _{1-6 alkylthio, -CN, -CF 3, -OR 3} , -COR 3, NO 2, -NHR 3, _{-NHC (O) R 3, -C} (O) NR 3 R 4, -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -OC (O) R 3, -C (O) R ₃ R ₄ , -O (CH ₂ ) _q NR ₃ , -CNR ₃ R ₄ , or - (CH ₂ ) _q NR ₃ R ₄ , wherein q is an integer from 1 to 6; n is 0, 1, 2, 3 or 4, and when n is > 1, R ₂ is the same or different; p is 0 or an integer from 1 to 5; Y is O, S, CHOH, -NHC ( O), -C (O) NH-, -C (O) -, -OC (O) -, NR 7 or -CN = N- and, R ₇ is H Or C _1-4 alkyl; Or lack thereof; Z is CR ₈ R ₉ , wherein R ₈ and R ₉ are each selected from H and straight or branched C _1-8 alkyl; Provided that - (CH ₂ ) _p -Y- is - (CH ₂ ) ₃ -C (O) - or - (CH ₂ ) ₃ -S-; X ₁ to X ₅ are C; -A- (R ₂ ) _n and R ₁ are each o-methyl-phenyl and n-butyl; Phenyl and n-butyl; Or each p- fluoro-phenyl, and is not a _{-O- (CH 2) 2 CH 3} ). The method according to claim 1, X ₁ , X ₂ , X ₃ , X ₄ and X ₅ are C; Or one of X ₁ , X ₂ , X ₃ , X ₄ or X ₅ is O or N and the others are all C; k is 0 or 1; t is 1; R ₁ is straight or branched C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _1-8 alkylidene, C _1-8 alkoxy, C _1-8 aminoalkyl, C _{1- 8} haloalkyl, C _{1-8 alkoxycarbonyl, -C (O) NR 3 R} 4, -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -OC (O) R 3 or - ( CH ₂ ) _s NR ₃ R ₄ wherein R ₃ , R ₄ and R ₅ are the same or different and are each selected from H and C _1-6 alkyl; s is an integer from 1 to 8; n is 1, 2 or 3; And A is phenyl or naphthyl; R ₂ is straight or branched C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _{1- 6 alkoxycarbonyl, -CN, -CF 3, -OH,} -COR 3, -NHR 3, -NHC (O) R 3, -C (O) NR 3 R 4, -NR 3 R 4, -NR 3 C (O) NR ₄ R ₅ , -OC (O) R ₃ or - (CH ₂ ) _q NR ₃ R ₄ wherein q is an integer from 1 to 6; or A is aryl containing at least one heteroatom selected from N, S and O; R ₂ is H, halo or a straight-chain or branched C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 heteroalkyl, C _1-6 aminoalkyl , C _1-6 haloalkyl, C _{1-6 alkoxycarbonyl, -CN, -CF 3, -OH,} -COR 3, -NHR 3, -NHC (O) R 3, -C (O) NR 3 R ₄ , -NR ₃ R ₄ , -NR ₃ C (O) NR ₄ R ₅ , -OC (O) R ₃ or - (CH ₂ ) _q NR ₃ R ₄ or a pharmaceutically acceptable salt thereof. Possible salts, esters or prodrugs. 3. The method according to claim 1 or 2, Wherein p is 3, or a pharmaceutically acceptable salt, ester or prodrug thereof. 3. The method according to claim 1 or 2, Wherein k is 0, or a pharmaceutically acceptable salt, ester or prodrug thereof. The method according to claim 1, Wherein said compound is a compound of formula < RTI ID = 0.0 > (II) < / RTI > or a pharmaceutically acceptable salt, ester or prodrug thereof. (II) 6. The method of claim 5, Wherein said compound is a compound of formula < RTI ID = 0.0 > (IIa) < / RTI > or a pharmaceutically acceptable salt, ester or prodrug thereof. (Formula IIa) The method according to claim 6, Wherein t is 1 and Y is -C (O) -, -NHC (O) -, S, O or -OC (O) -, or a pharmaceutically acceptable salt, ester or prodrug thereof. Drug. 8. The method of claim 7, Wherein X < ₃ > is C, or a pharmaceutically acceptable salt, ester or prodrug thereof. 9. The method of claim 8, Wherein R < ₁ > is alkyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 10. The method of claim 9, Wherein R < ₂ > is alkyl, aminoalkyl, alkoxy or hydroxyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 11. The method of claim 10, Wherein p is 3, or a pharmaceutically acceptable salt, ester or prodrug thereof. 12. The method of claim 11, _Wherein R ₁ is C _2-8 alkyl and R ₂ is methyl, hydroxyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 13. The method of claim 12, Wherein Y is -C (O) - or O, or a pharmaceutically acceptable salt, ester or prodrug thereof. 10. The method of claim 9, Lt; ₂ > is halo, or a pharmaceutically acceptable salt, ester or prodrug thereof. The method according to claim 6, Wherein t is 0, or a pharmaceutically acceptable salt, ester or prodrug thereof. 9. The method of claim 8, Wherein R < ₁ > is alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 9. The method of claim 8, Wherein R < _{1 >} is benzyl or phenyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 8. The method of claim 7, Wherein X < ₃ > is N, or a pharmaceutically acceptable salt, ester or prodrug thereof. 19. The method of claim 18, Wherein R < ₁ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 19. The method of claim 18, Wherein R < _{1 >} is benzyl or phenyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 20. The method of claim 19, Wherein R < ₂ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 21. The method of claim 20, Wherein R < ₂ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 8. The method of claim 7, Wherein X < ₃ > is O, or a pharmaceutically acceptable salt, ester or prodrug thereof. 24. The method of claim 23, Wherein R < ₁ > is alkyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 25. The method of claim 24, Wherein R < ₂ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 25. The method of claim 24, Lt; ₂ > is halo, or a pharmaceutically acceptable salt, ester or prodrug thereof. 6. The method of claim 5, Wherein said compound is a compound of formula < RTI ID = 0.0 > (IIb) < / RTI > or a pharmaceutically acceptable salt, ester or prodrug thereof. (IIb) 28. The method of claim 27, Wherein Y is -C (O) -, -NHC (O) -, S, O or -OC (O) -; or a pharmaceutically acceptable salt, ester or prodrug thereof. 29. The method of claim 28, Wherein X < ₃ > is C, or a pharmaceutically acceptable salt, ester or prodrug thereof. 30. The method of claim 29, Wherein R < ₁ > is alkyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 31. The method of claim 30, Wherein R < ₂ > is alkyl, aminoalkyl, alkoxy or hydroxyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 32. The method of claim 31, Wherein p is 3, or a pharmaceutically acceptable salt, ester or prodrug thereof. 33. The method of claim 32, _Wherein R ₁ is C _2-8 alkyl and R ₂ is methyl, hydroxyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 34. The method of claim 33, Wherein Y is -C (O) - or O, or a pharmaceutically acceptable salt, ester or prodrug thereof. 31. The method of claim 30, Lt; ₂ > is halo, or a pharmaceutically acceptable salt, ester or prodrug thereof. 28. The method of claim 27, Wherein t is 0, or a pharmaceutically acceptable salt, ester or prodrug thereof. 30. The method of claim 29, Wherein R < ₁ > is alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 30. The method of claim 29, Wherein R < _{1 >} is benzyl or phenyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 29. The method of claim 28, Wherein X < ₃ > is N, or a pharmaceutically acceptable salt, ester or prodrug thereof. 40. The method of claim 39, Wherein R < ₁ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 40. The method of claim 39, Wherein R < _{1 >} is benzyl or phenyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 41. The method of claim 40, Wherein R < ₂ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 42. The method of claim 41, Wherein R < ₂ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 29. The method of claim 28, Wherein X < ₃ > is O, or a pharmaceutically acceptable salt, ester or prodrug thereof. 45. The method of claim 44, Wherein R < ₁ > is alkyl, or a pharmaceutically acceptable salt, ester or prodrug thereof. 46. The method of claim 45, Wherein R < ₂ > is alkyl or alkoxy, or a pharmaceutically acceptable salt, ester or prodrug thereof. 46. The method of claim 45, Lt; ₂ > is halo, or a pharmaceutically acceptable salt, ester or prodrug thereof. Characterized in that the muscarinic receptor is contacted with an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, ester or prodrug thereof. (I) Wherein X ₁ , X ₂ , X ₃ , X ₄ and X ₅ are selected from C, N and O; k is 0 or 1; t is 0 or 1; R ₁ is straight or branched C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _1-8 alkylidene, C _1-8 alkoxy, C _1-8 heteroalkyl, C _{1- 8} aminoalkyl, C _1-8 haloalkyl, C _1-8 alkoxycarbonyl, C _1-8 hydroxy alkoxy, C _1-8 hydroxyalkyl, -SH, C _1-8 thioalkyl, -O-CH ₂ -C (O) -C _5-6 aryl substituted with C _5-6 aryl, C _1-3 alkyl or halo; C _5-6 aryl or C _5-6 cycloalkyl optionally containing one or more heteroatoms selected from N, S and O (89620); _{-C (O) NR 3 R 4} , -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -CR 3 R 4, -OC (O) R 3, -C (O) (CH 2 ) _s NR ₃ R ₄ or - (CH ₂ ) _s NR ₃ R ₄ wherein R ₃ , R ₄ and R ₅ are the same or different and each is H, C _1-6 alkyl, N, O and S selected and, optionally, halo, or C _5-6 aryl optionally including one or more characters of heteroatom substituted with C _1-6 alkyl in; and the N atom, or R ₃ and R _4; selected from C _3-6 cycloalkyl, and If present, forms a cyclic ring structure comprising 5-6 atoms selected from C, N, S and O; s is an integer from 0 to 8; A is N, C, including the at least one heteroatom selected from S and O optionally _5-12 aryl or C _5-7 cycloalkyl; R ₂ is H, amino, hydroxyl, halo, or straight or branched C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 heteroalkyl, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _{1-6 alkoxycarbonyl, -CN, -CF 3, -OR 3} , -COR 3, NO 2, -NHR 3, -NHC (O) R 3 _{, -C (O) NR 3 R} 4, -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -OC (O) R 3, -C (O) R 3 R 4, -O ( CH ₂ ) _q NR ₃ , -CNR ₃ R ₄ , or - (CH ₂ ) _q NR ₃ R ₄ (wherein q is an integer from 1 to 6); n is 0, 1, 2, 3 or 4, and when n is > 1, R ₂ is the same or different; p is 0 or an integer from 1 to 5; Y is O, S, CHOH, -NHC ( O), -C (O) NH-, -C (O) -, -OC (O) -, NR 7 or -CN = N- and, R ₇ is H Or C _1-4 alkyl; Or lack thereof; Z is CR ₈ R ₉ , wherein R ₈ and R ₉ are each selected from H and straight or branched C _1-8 alkyl. 49. The method of claim 48, Wherein said compound is a compound of formula < RTI ID = 0.0 > (II) < / RTI > (II) 50. The method of claim 49, Wherein t is 1 and Y is -C (O) -, -NHC (O) -, S, O or -OCO-. 51. The method of claim 50, Wherein X ₃ is a method of reacting a muscarinic receptor, characterized in that C. 52. The method of claim 51, RTI ID = 0.0 > R1 < / RTI _> is alkyl. 53. The method of claim 52, Wherein said R < ₂ > is alkyl, aminoalkyl, alkoxy or hydroxyl. 54. The method of claim 53, Lt; RTI ID = 0.0 > p is 3. < / RTI > 55. The method of claim 54, _Wherein R ₁ is C _2-8 alkyl and R ₂ is methyl, hydroxyl or alkoxy. 56. The method of claim 55, Wherein Y is -C (O) - or < RTI ID = 0.0 > O. < / RTI > 53. The method of claim 52, RTI ID = 0.0 > R2 < / RTI _> is halo. 50. The method of claim 49, Lt; RTI ID = 0.0 > 0 < / RTI > 52. The method of claim 51, RTI ID = 0.0 > R1 < / RTI _> is alkoxy. 52. The method of claim 51, Wherein R < _{1 >} is benzyl or phenyl. 53. The method of claim 52, Wherein X ₃ is a method of reacting a muscarinic receptor, characterized in that N. 62. The method of claim 61, RTI ID = 0.0 > R1 < / RTI _> is alkyl or alkoxy. 62. The method of claim 61, Wherein R < _{1 >} is benzyl or phenyl. 63. The method of claim 62, Wherein R ₂ is a method of reacting a muscarinic receptor, characterized in that alkyl or alkoxy. 64. The method of claim 63, Wherein R ₂ is a method of reacting a muscarinic receptor, characterized in that alkyl or alkoxy. 51. The method of claim 50, The method for X ₃ are acting muscarinic receptor, characterized in that O. 67. The method of claim 66, RTI ID = 0.0 > R1 < / RTI _> is alkyl. 68. The method of claim 67, Wherein R ₂ is a method of reacting a muscarinic receptor, characterized in that alkyl or alkoxy. 68. The method of claim 67, RTI ID = 0.0 > R2 < / RTI _> is halo. Claims 1. A pharmaceutical composition comprising an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt, ester or prodrug thereof, and a pharmaceutically acceptable carrier. (I) Wherein X ₁ , X ₂ , X ₃ , X ₄ and X ₅ are selected from C, N and O; k is 0 or 1; t is 0 or 1; R ₁ is straight or branched C _1-8 alkyl, C _2-8 alkenyl, C _2-8 alkynyl, C _1-8 alkylidene, C _1-8 alkoxy, C _1-8 heteroalkyl, C _{1- 8} aminoalkyl, C _1-8 haloalkyl, C _1-8 alkoxycarbonyl, C _1-8 hydroxy alkoxy, C _1-8 hydroxyalkyl, -SH, C _1-8 thioalkyl, -O-CH ₂ -C (O) -C _5-6 aryl substituted with C _5-6 aryl, C _1-3 alkyl or halo; C _5-6 aryl or C _5-6 cycloalkyl optionally containing one or more heteroatoms selected from N, S and O; _{-C (O) NR 3 R 4} , -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -CR 3 R 4, -OC (O) R 3, -C (O) (CH 2 ) _s NR ₃ R ₄ or - (CH ₂ ) _s NR ₃ R ₄ wherein R ₃ , R ₄ and R ₅ are the same or different and each is H, C _1-6 alkyl, N, O and S selected and, optionally, halo, or C _5-6 aryl optionally including one or more characters of heteroatom substituted with C _1-6 alkyl in; and the N atom, or R ₃ and R _4; selected from C _3-6 cycloalkyl, and If present, forms a cyclic ring structure comprising 5-6 atoms selected from C, N, S and O; s is an integer from 0 to 8; A is N, C, including the at least one heteroatom selected from S and O optionally _5-12 aryl or C _5-7 cycloalkyl; R ₂ is H, amino, hydroxyl, halo, or straight or branched C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _1-6 alkoxy, C _1-6 heteroalkyl, C _1-6 aminoalkyl, C _1-6 haloalkyl, C _{1-6 alkoxycarbonyl, -CN, -CF 3, -OR 3} , -COR 3, NO 2, -NHR 3, -NHC (O) R 3 _{, -C (O) NR 3 R} 4, -NR 3 R 4, -NR 3 C (O) NR 4 R 5, -OC (O) R 3, -C (O) R 3 R 4, -O ( CH ₂ ) _q NR ₃ , -CNR ₃ R ₄ , or - (CH ₂ ) _q NR ₃ R ₄ (wherein q is an integer from 1 to 6); n is 0, 1, 2, 3 or 4, and when n is > 1, R ₂ is the same or different; p is 0 or an integer from 1 to 5; Y is O, S, CHOH, -NHC ( O), -C (O) NH-, -C (O) -, -OC (O) -, NR 7 or -CN = N- and, R ₇ is H Or C _1-4 alkyl; Or lack thereof; Z is CR ₈ R ₉ , wherein R ₈ and R ₉ are each selected from H and straight or branched C _1-8 alkyl. 71. The method of claim 70, Wherein said compound is a compound of formula < RTI ID = 0.0 > (II) < / RTI > (II) 72. The method of claim 71, Wherein t is 1 and Y is -C (O) -, -NHC (O) -, S, O or -OC (O) -. 73. The method of claim 72, Wherein X < ₃ > is C; 77. The method of claim 73, Wherein R < ₁ > is alkyl. 75. The method of claim 74, Wherein said R < ₂ > is alkyl, aminoalkyl, alkoxy or hydroxyl. 78. The method of claim 75, Wherein p is 3. < RTI ID = 0.0 > 8. < / RTI > 78. The method of claim 75, _Wherein R ₁ is C _2-8 alkyl and R ₂ is methyl, hydroxyl or alkoxy. 78. The method of claim 77, Wherein Y is -C (O) - or O. < RTI ID = 0.0 > 75. The method of claim 74, Wherein said R < ₂ > is halo. 72. The method of claim 71, Wherein t is 0. 77. The method of claim 73, Wherein R < ₁ > is alkoxy. 77. The method of claim 73, Wherein R < _{1 >} is benzyl or phenyl. 75. The method of claim 74, Wherein X < ₃ > is N. 83. The method of claim 82, Wherein R < ₁ > is alkyl or alkoxy. 83. The method of claim 82, Wherein R < _{1 >} is benzyl or phenyl. 85. The method of claim 83, Wherein R < ₂ > is alkyl or alkoxy. 85. The method of claim 84, Wherein R < ₂ > is alkyl or alkoxy. 73. The method of claim 72, Wherein X < ₃ > is O; 90. The method of claim 88, Wherein R < ₁ > is alkyl. 90. The method of claim 89, Wherein R < ₂ > is alkyl or alkoxy. 88. The method of claim 87, Wherein said R < ₂ > is halo. 11. A method of treating a symptom of a disease or disorder by the reduction of acetylcholine, which comprises administering a therapeutically effective amount of at least one compound of claim 1. 93. The method of claim 92, Wherein said disease or disorder is a neuronal disease, cognitive impairment, cognitive decline due to age or dementia. A method of treating a symptom of a disease or disorder by increasing the intraocular pressure, characterized by administering a therapeutically effective amount of at least one compound of claim 1. 95. The method of claim 94, Wherein the disease is glaucoma. ≪ RTI ID = 0.0 > 11. < / RTI > A method of treating a symptom of a disease or disorder caused by a reduction in acetylcholine, the method comprising administering a therapeutically effective amount of the composition of claim 70. 96. The method of claim 96, Wherein said disease or malignant neurogenic disease, cognitive impairment, cognitive decline by age or dementia is characterized by the reduction of acetylcholine. A method of treating a symptom of a disease or disorder by increasing the intraocular pressure, characterized by administering a therapeutically effective amount of the composition of claim 70. 98. The method of claim 98, Wherein the disease is glaucoma. ≪ RTI ID = 0.0 > 11. < / RTI >