WO2001002386A1 - SELECTIVE M2 MUSCARINIC RECEPTOR ANTAGONISTS HAVING 5H-DIBENZ[b,f]AZEPINE STRUCTURE - Google Patents

Abstract

Novel 5H-dibenzo[b,f]azepine derivatives of general formula (I) wherein X-Y represents CH2-CH2, CH=CH, CH=CR3; R1 represents hydrogen or linear or branched C1-4 alkyl; R2 represents linear or branched C1-4 alkyl, phenyl, benzyl or phenethyl; R3 represents hydroxyl, linear or branched C1-4 alkoxy, phenoxyl; m and n are independently an integer 1 to 10, are disclosed. These compounds are capable of selectively blocking muscarine M2 receptors and can be used in the treatment of cardiovascular disorders, particularly bradycardias and bradyarrhythmias and in the treatment of cognitive disorders such as Alzheimer's disease.

Description

"SELECTIVE M₂ MUSCARINIC RECEPTOR ANTAGONISTS HAVING 5H-DIBENZMAZEPINE STRUCTURE"

FIELD OF THE INVENTION

The present invention relates to novel pharmacologically active 5H-dibenz[b_j ]azepine derivatives, the pharmaceutical compositions containing them and the use of such compounds/compositions for the treatment of cardiovascular disorders, more specifically bradycardia and brady arrhythmia and for treatment of cognitive disorders such as Alzheimer's disease.

TECHNOLOGICAL BACKGROUND

Muscarine cholinergic receptors are present in a number of tissues. The activation of these receptors through their neurotransmitter, acetylcholine, can elicit different effects such as the increase in gastrointestinal motility, miosis, increase in the salivary and intestinal secretions, eminction stimulus, bradycardia. More particularly, M2 receptors are present in the cardiac tissue at post-synaptic level, and in hippocampus and cerebral cortex at pre-synaptic level (inhibitor autorececeptors).

Post-synaptic M2 receptors play a paramount role in the regulation of vagus nerve-mediated heart rate. Hyperstimulation of M2 receptors causes an increase in the parasympathetic activity and therefore is apparently an important risk factor in the sick sinus syndrome and in atrioventricular block. IV receptor antagonists may therefore be used in the treatment of atrial sinus functional disorders. Compounds with this type of activity are known, such as AF-DX 1 16

(US 4550107), AQ-RA 741 (EP 312895), YM 47244 (WO 9613488) but the only commercially available product is atropine, a non-selective muscarine antagonist.

The use of this product is however restricted by its short-lasting action and by the onset of side effects, due to its poor selectivity, such as fauces dryness, mydriasis, decreased gastrointestinal motility. Therefore, there is need for compounds selective towards M2 receptors having long-lasting action and devoid of side effects.

Pre-synaptic M2 receptors modify the acetylcholine release through a negative feedback mechanism. The release of endogenous acetylcholine can be promoted by blocking these receptors with selective antagonists. The increase in acetylcholine can improve cognitive performance under hypofunctionality conditions of the cholinergic system as is the case with Alzheimer's disease. In this field, acetylcholinesterase inhibitors such as tacrine (New Engl. J. Med.

315,1241-5 (1986)), donepezil (EP 296560), rivastigmine (DE 3805744), heptastigmine (EP 154864) have been widely studied and used but not altogether successfully in that they induce even severe side effects. Tacrine induces high levels of transaminases (JAMA 280, 1777-1782 (1998)) whereas heptastigmine can cause serious hematologic toxic effects (Neurology 52, (4), 700-708 (1999)).

The present invention aimed at finding muscarine M2 receptors antagonists capable of promoting the cholinergic central transmission without inducing serious side effects.

SUMMARY OF THE INVENTION

The present invention relates to a novel class of 5H-dibenzo[b,fJazepine derivatives having high activity and selectivity for muscarine M2 receptors, long half-life, and without side effects. The novel compounds are potentially useful in the treatment of cardiovascular disorders, in particular in the treatment of sinus bradycardia, sick sinus syndromes, disorders of atrio-ventricular conduction and bradyarrhythmias, or in the treatment of cognitive and neurodegenerative disorders such as Alzheimer's disease. These novel derivatives have the general formula (I)

( I )

wherein X-Y represents CH₂-CH₂, CH-CH, CH=CR3;

R\ represents hydrogen or linear or branched Cι_4 alkyl;

R2 represents linear or branched Cι_4 alkyl, phenyl, benzyl or phenethyl; R3 represents hydroxyl, linear or branched C 1.4 alkoxy, phenoxyl; m and n are independently an integer 1 to 10.

In a preferred group of compounds, X-Y is CH2-CH2 and the alkylamino group is substituted at the piperidine at the 2- position.

In another preferred group of compounds, X-Y is CH2-CH2 and the alkylamino group is substituted at the piperidine at the 3- position.

In another preferred group of compounds, X-Y is CH2-CH2 and the alkylamino group is substituted at the piperidine at the 4- position.

In another preferred group of compounds, X-Y is CH=CH and the alkylamino group is substituted at the piperidine at the 2- position. In another preferred group of compounds, X-Y is CH=CH and the alkylamino group is substituted at the piperidine at the 3- position.

In another preferred group of compounds, X-Y is CH=CH and the alkylamino group is substituted at the piperidine at the 4- position. In another preferred group of compounds, X-Y is CH=CR3 and the alkylamino group is substituted at the piperidine at the 2- position.

In another preferred group of compounds, X-Y is CH=CR3 and the alkylamino group is substituted at the piperidine at the 3- position.

In another preferred group of compounds, X-Y is CH=CR3 and the alkylamino group is substituted at the piperidine at the 4- position.

The compounds of formula (I) can be salified with pharmacologically acceptable inorganic or organic acids selected, for example, from hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, citric acid, oxalic acid, maleic acid, fumaric acid, tartaric acid, malonic acid, gluconic acid, salicylic acid, succinic acid, lactic acid and the like. DETAILED DESCRIPTION OF THE INVENTION

The characteristics and the advantages of the 5H-dibenz[b_j/]azepine derivatives suitable as selective M2 muscarinic receptor antagonists according to the present invention, and also the process for their preparation, will be mainly pointed out in the course of the following detailed description.

The compounds of the present invention have the following general formula:

wherein

X-Y represents CH2-CH2, CH=CH, CH=CR3; R\ represents hydrogen or a linear or branched Cι _4 alkyl; R2 represents a linear or branched Cj_4 alkyl, phenyl, benzyl or phenylethyl; R3 represents hydroxyl, linear or branched Cj_4 alkoxyl, phenoxyl; m and n, are independently an integer 1 to 10.

The novel 5H-dibenz[b_j ]azepine derivatives having formula (I) can be prepared by reacting of haloacyl derivatives (II), wherein Hal can be chlorine, bromine or iodine, with an amine of formula (III), according to the following process:

The amination reaction is carried out in an inert solvent at a temperature between 0°C and the boiling temperature of the solvent. The amine (II) is typically present in a two fold excess, or in one or two fold excess and in the presence of an auxiliary base. The reaction may be carried out in chlorinated solvents such as dichloromethane, chloroform, dichloroethane; aromatic solvents such as benzene, toluene, chlorobenzene, pyridine; ketones such as acetone, acetonitrile, dimethylformarnide; alcohols such as methanol, ethanol, isopropanol. The auxiliary base may be an organic base such as triethylamine, N,N-dimethylaniline, pyridine, 4-(dimethylamino)pyridine, or a organic base such as carbonate or hydrogen carbonate of alkali and alkaline earth metals. The reaction may be accelerated by adding catalytic amounts of alkali metal iodides. The reaction is typically carried out for a period of 30 minutes to 10 hours, depending on the compound (I) and amine (III) used.

Compounds of formula (II), in which X-Y is CH2-CH2, m=l and Hal is chlorine are described in J Med Chem 1987; 30: 1378-1382 and in GB Patent No. 849,032

(Geigy), respectively.

The other compounds of formula (II) may be prepared by reacting a

5H-dibenz[b_j ]azepine derivative (IV) with a compound of formula (V), in which

Hal and Hal', which are the same or different, can be chlorine, bromine or iodine.

( II)

The acylation is carried out in an inert solvent, at a temperature between room temperature and the boiling temperature of the solvent, preferably in the presence of an auxiliary base. The reaction may be carried out in chlorinated solvents such as dichloromethane, chloroform, dichloroethane; aromatic solvents such as benzene, toluene, chlorobenzene; cyclic or acyclic ethers such as diisopropyl ether, tetrahydrofuran, dioxane. The auxiliary base may be an organic base such as tnethylamine, N,N-dimethylaniline, pyridine, 4-(dimethylammo)pyridine, or an organic base such as carbonate or hydrogen carbonate of alkali and alkaline earth metals.

Compounds of formula (IV) in which X-Y is CH=COMe or CH=COEt are described in Synthetic Comm 1994; 24: 683-687. The compound of formula (IV) in which X-Y is CH=COnBu, is described in GB Patent No. 943,277 (Geigy). Compounds of formula (IV) in which X-Y is CH=COPh, is prepared from 5-acetyl-10-bromo-5H-dibenz[b_j ]azepine (VI), disclosed in GB Patent No. 943,277 (Geigy), according the following procedure:

(VIII)

The compound having formula (VI) is reacted with phenol (VII), at a temperature of 100°C, in dimethylsulfoxide as solvent. The hetaryne intermediate formed by dehydrohalogenation of (VT) reacts with phenol (VTI) to give 10-phenoxy- 5H-dibenz[b, ]azepine (VIII).

A number of amines having formula (III) are reported in literature. For example, the amines (III) in which n=l and R] and R2 are alkyl groups are disclosed in US Patent No. 4,550,107 (Karl Thomae); the amine (III) in which n=2 and Rι=R2=Et is reported in J Am Chem Soc 1957; 79: 2836-2838; the amines (III) in which n=3-5 and Rj and R2 are alkyl groups are described in Liebigs Ann Chem 1993: 809-810. The 4-[4-(diethylamino)butyl]piperidine (XV), described in literature in Liebigs Ann Chem 1993: 809-810, was synthesized with a novel method reported hereinbelow: COOEt

(XII ) (xiii) (XIV) (XV)

The N-benzyl-piperidone is treated with triethyl phosphonocrotonate (Horner-Emmons reaction) and sodium ethoxide in ethanol to afford compound (IX). This compound is hydrogenated, with 10% palladium on activated carbon, under atmospheric pressure and room temperature in ethanol to give (X). The compound (X) is benzylated with benzyl chloride to give (XI), then reduced with lithium aluminum hydride in tetrahydrofuran at room temperature to afford alcohol (XII). Conversion of alcohol (XII) to the corresponding methanesulfonyl derivative (XIII), with methanesulfonyl chloride at 0°C in tetrahydrofuran, followed by reaction with two equivalents of diethylamine, in acetonitrile at 60°C, gives (XIV). The N-debenzylation of (XIV) with 10% palladium on activated carbon and ammonium formate in methanol under reflux gives the 4-[4-(diethylamino)butyl]piperidine (XV).

The 4-[4-(N-benzyl-N-methylamino)butyl]piperidine (XIX) is prepared from methanesulfonyl derivative (XIII) with the following procedure.

(XIII) (XVI) (XVII)

(XVIII) (XIX)

The methanesulfonyl derivative (XIII) is treated with methylamine 40% in water in acetonitrile at room temperature to give amino derivative (XVI). This compound is benzoylated with benzoyl chloride and sodium hydroxide in acetone at room temperature to give amide (XVII). The N-debenzylation of (XVII) with 10% palladium on activated carbon and ammonium formate in methanol at reflux gives (XVIII). The reduction of (XVIII) with lithium aluminum hydride in tetrahydrofuran at reflux gives 4-[4-(N-benzyl-N-methylamino)butyl]piperidine (XIX).

The 4-[4-(diethylamino)heptyl]piperidine (XXIII) is prepared from 4-picoline and 6-bromohexanoyl chloride with the following procedure.

Br(CH₂)₅CONEt₂

(XXI) (XXII) (XXIII) (xx>

Br(CH₂)₅COCI

The amide (XX) is obtained by reacting 6-bromohexanoyl chloride and diethyl amine in acetonitrile at room temperature. Compound (XX) is treated with 4-picolyl sodium, obtained from 4-picoline and sodium amide 50% in toluene, to give (XXI). The hydrochloride of (XXI), obtained by treatment with a solution of hydrogen chloride in methanol, is hydrogenated with 5% platinum oxide under atmospheric pressure and room temperature, in acetic acid to give (XXII). The reduction of (XXII) with lithium aluminum hydride in tetrahydrofuran under reflux gives 4-[4-(diethylamino)heptyl]piperidine (XXHl). The present invention provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, as active ingredient, in association with a pharmaceutical acceptable carrier, excipient or other additive, if necessary. The pharmaceutical compositions containing a compound of formula (I) can be administered, for example, through the oral, parenteral, rectal, transdermal routes, in the form of capsules, tablets, solutions, suppositories, ointments and the like. The daily dosage will depend on a number of factors, such as the type of disease and the conditions of the patient (weight, sex and age) and it will be determined according to standard procedures on the basis of the pharmacokinetic and toxicological characteristics of each compound. An average daily dosage will typically comprise 0.1 to 10 mg/kg. In order to illustrate the preparation of the compounds according to the invention the following Examples are reported. EXAMPLE 1 a) Ethyl i'r «s-4-(N-Benzyl-4'-piperidinyliden)-2-butenoate (IX) l-Benzyl-4-piperidone (17.1 mL, 92.2 mmol) and triethyl

4-phosphonocrotonate (25.63 g, 92.2 mmol) were dissolved in 60 mL of absolute ethanol and cooled to 0 - 5°C, under a nitrogen atmosphere. Metal sodium (2.76 g, 119.9 mmol) dissolved in 150 mL of absolute ethanol was added dropwise to the cooled solution at below 5°C during 40 min. The solution was stirred at 5°C for 45 min and 1 h at room temperature. The reaction mixture was diluted with 600 mL of brine and extracted with diethyl ether (5 x 100 mL). The combined extract was washed with water, dried over Na2SC>4, the solvent was evaporated off in vacuo and the residue purified by flash column chromatography eluting with petroleum ether-Et2θ (1: 1, v/v) to afford 16.6 g of (IX) as a yellow oil. Yield 63%. IR (neat) cm-1 ; 2920, 2800, 1710 (CO), 1635, 1610. iH-NMR (CDCI3) δ (ppm): 1.29 (3H, t, J=7.0 Hz), 2.25 - 2.60 (8H, m), 3.51 (2H, s), 4.19 (2H, q, J=7.0 Hz), 5.80 (IH, d, J=15.2 Hz, CH-COOEt), 5.96 (IH, d, J=10.9 Hz, CH-CHOH-COOEt), 7.20 - 7.35 (5H, m), 7.56 (IH, dd, J=15.2, 10.9 Hz, CHOH-COOEt). b) Ethyl 4-(4'-piperidinyl)butanoate (X)

A suspension of (IX) (16.48 g, 57.7 mmol) and 1.6 g of 10% palladium on activated carbon in 150 mL of ethanol was hydrogenated at atmospheric pressure and room temperature for 8 h. The reaction mixture was filtered through Celite® and the solvent was evaporated off under reduced pressure. The residue was dissolved in 100 mL of diethyl ether and the solvent was washed with brine, dried over Na2Sθ4, filtered and evaporated in vacuo to give 11.2 g of (X) as a yellow oil. Yield 97%.

IR (neat) cm"¹: 3280 (NH), 2910, 2840, 1725 (CO).

!H-NMR (CDCI3) δ (ppm): 1.22 (3H, t, J=7.0 Hz), 1.55 - 1.78 (9H, m), 2.25 (2H, t, J=6.8 Hz), 2.53 (2H, td, J-11.6, 2.6 Hz, NCHax), 3.01 (2H, d, J=1 1.6 Hz, NCHgq), 4.09 (2H, q, J=6.8 Hz). c) Ethyl 4-(N-Benzyl-4'-piperidinyl)butanoate (XI)

Sodium carbonate (5.9 g, 55.4 mmol), benzyl chloride (6.4 mL, 55.4 mmol) and a catalytic amount of sodium iodide were added to (X) (11.04 g, 55.4 mmol) dissolved in 60 mL of dimethylformamide. The reaction mixture was stirred at room temperature for 6 h, diluted in 400 mL of 0.1 N HCl and extracted with diethyl ether (3 x 80 mL). The aqueous layer was basified with 40% aqueous NaOH and extracted with diethyl ether (4 x 100 mL). The combined organic phases were washed with water, dried over Na2S04, filtered and evaporated to give 13.85 g of (XI) as yellow oil. Yield 86%. IR (neat) cm^~l : 2927, 2840, 2800, 1736 (CO), 1454. iH-NMR (CDCI3) δ (ppm): 1.15 - 1.35 (8H, m), 1.50 - 1.70 (4H, m), 1.91 (2H, t br J=10.4 Hz, NCH_ax), 2.27 (2H, t, J=7.1 Hz), 2.86 (2H, d, J=11.2 Hz, NCH_eq), 3.46 (2H, s, CH₂Ph), 4.11 (2H, q, J=6.8 Hz), 7.20 - 7.35 (5H, m). d) 4-(N-Benzyl-4'-piperidinyl)-l-butanol (XII) A solution of (XI) (13.8 g, 47.8 mmol) in 60 mL of anhydrous tetrahydrofuran was added dropwise, under a nitrogen atmosphere, to a suspension of lithium aluminum hydride (2.7 g, 71.7 mmol) in 40 mL of anhydrous tetrahydrofuran while maintaining the reaction temperature below 10°C. The mixture was stirred for 1 h at room temperature, cooled to 0°C and then hydrolyzed by addition of 1 N NaOH. The resulting suspension was filtered and the filtrate evaporated. The residue was dissolved in 100 mL of diethyl ether and washed with water and the organic layer was dried over Na2S04, filtered and evaporated in vacuo to give 11.3 g of (XII) as yellow oil. Yield 96%. IR (neat) cm"¹: 3339 (O-H), 2928, 1454.

1H-NMR (CDCI3) δ (ppm): 1.10 - 1.70 (11H, m), 1.91 (2H, t br J=l l.l Hz, NCH_ax), 2.86 (2H, d, J=10.9 Hz, NCH_eq), 3.47 (2H, s, CH₂Ph), 3.61 (2H, t, J=5.7 Hz), 7.20 - 7.35 (5H, m). e) 4-[4-(Diethylamino)butyI]-N-benzylpiperidine (XIV)

Triethylamine (6.4 ml, 45.8 mmol) and (XII) (1 1.23 g, 45.4 mmol) were dissolved in 60 mL of tetrahydrofuran and cooled to 0°C. Methanesulfonyl chloride (3.6 mL, 45.8 mmol) dissolved in 20 mL of tetrahydrofuran was added dropwise, while maintaining the reaction temperature below 5°C. The mixture was stirred for 45 min at this temperature and then warmed to room temperature. The resulting suspension was diluted with 60 mL of diethyl ether and the insoluble precipitate was filtered off. The solvent was evaporated in vacuo to give 15.20 g of (XIII) as a yellow oil. The compound was characterized only by IR and reacted immediately. IR (neat) cm^{" 1} : 2928, 1454,1355 (v_as S0₂), 1176 (v_s SO2). Methanesulfonate derivative (XIII) (15.03 g, 46.2 mmol) and diethylamine (19.1 mL, 184.8 mmol) dissolved in 130 mL of acetonitrile were refluxed for 6 h. After cooling to room temperature the mixture was diluted with 250 mL of brine and extracted with diethyl ether (4 x 80 mL). The combined organic phases were washed with water, dried over Na2S04, filtered and evaporated in vacuo to give 12.94 g of (XIV) as brownish oil. Yield 94%. IR (neat) cm-l; 2928, 2840, 2790, 1454.

1H-NMR (CDCI3) δ (ppm): 0.99 (6H, t, J=7.0 Hz), 1.10 - 1.70 (11H, m), 1.91 (2H, m, NCH_ax), 2.38 (2H, t, J=8.4 Hz), 2.50 (4H, q, J=7.0 Hz), 2.85 (2H, d br, J=10.0 Hz, NCH_eq), 3.46 (2H, s, CH₂Ph), 7.20 - 7.35 (5H, m). f) 4-[4-(Diethylammo)butyl]piperidine (XV)

A mixture of (XIV) (12.9 g, 42.7 mmol), ammonium formate (13.5 g, 213.5 mmol) and 10% palladium on activated carbon (13 g) in 160 mL of anhydrous methanol was refluxed for 30 min under nitrogen atmosphere. After cooling to room temperature the reaction mixture was filtered through Celite® and the solvent evaporated off to afford 8.13 g of (XV) as yellow oil. Yield 90%. IR (neat) cm-¹ : 3278 (N-H), 2929, 1466, 1446. iH-NMR (CDCI3) δ (ppm): 0.99 (6H, t, J=7.1 Hz), 1.05 - 1.75 (12H, m), 2.38 (2H, t, J=8.1 Hz), 2.50 (4H, q, J=7.1 Hz), 2.55 (2H, m, NCH_ax), 3.02 (2H, dt, J=12.2, 2.8 Hz, CH_eq). g) 5-{ [4-[4-(Diethylamino)butyl]-l-piperidinyl}acetyl-5H- dibenz[b,f]azepine Sodium carbonate (1.36 g, 12.8 mmol), 5-(chloroacetyl)-5H-dibenz[b, ]azepine (3.46 g, 12.8 mmol) and a catalytic amount of sodium iodide were added to (XV) (3 g, 14.1 mmol) in 70 mL of acetonitrile. The mixture was refluxed for 1.5 h, diluted in 150 mL of 2 N HCl and extracted with diethyl ether (2 x 80 mL). The aqueous layer was basified with 40% aqueous NaOH and extracted with diethyl ether (3 x 80 mL). The combined organic phases were washed with water, dried over Na2Sθ4, filtered and concentrated in vacuo. The residue was purified by flash column chromatography eluting with petroleum ether-acetone-triethyl amine (16:4:0.5, v/v/v). After evaporating the solvent, the residue was dissolved in diethyl ether and washed with water (3 x 100 mL). The organic layer was dried over Na2S04, filtered and evaporated in vacuo to give 5.04 g of 5-{[4-[4- (diethylamino)butyl]-l-piperidinyl}acetyl-5H-dibenz[b_j ]azepine as a light yellow oil. Yield 88%.

Elemental Analysis C H N

(theor. %) 78.16 8.82 9.43

(found %) 78.06 8.90 9.37 IR (neat) cm-l; 3024, 2929, 1676 (CO), 1490, 1462, 1442. iH-NMR (CDCI3) δ (ppm): 0.99 (6H, t, J=6.9 Hz, NCH₂CH ), 1.05 - 2.00 (13H, m, CH₂, CH and NCH_ax), 2.37 (2H, m, CH₂NEt₂), 2.50 (4H, q, J=6.5 Hz, NCH₂CH₃), 2.68 (2H, d, J=9.8, NCH_eq). 2.76 (IH, d, J=14.3 Hz, COCHH'), 3.07 (IH, d, J=14.3 Hz, COCHH'), 6.91 (2H, dd, J=13.7, 11.7 Hz, CHOH), 7.30 -

7.50 (8H, m).

EXAMPLE 2 a) 4-[4-(Methylamino)butyI]-N-beιιzylpiperidine (XVI) To a stirred solution of methanesulfonate derivative (XIII) (1.65 g, 5.1 mmol) in acetonitrile (7 mL) was added methylamine 40% in water (102 mmol) and the mixture was stirred for 15 h at room temperature. The reaction was poured into brine (80 mL), extracted with diethyl ether (3 x 50 mL), dried over Na₂Sθ4 and the solvent was removed in vacuo to afford 1.35 g of (XVI) as a yellow oil. Yield 98%.

IR (neat) cm"¹: 3250 (NH), 2900, 2830, 2780, 1440.

1H-NMR (CDCI3) δ (ppm): 1.14 - 1.70 (11H, m), 1.90 (2H, t br, J-11.5 Hz, NCHax), 2.41 (3H, s), 2.54 (2H, J=7.1 Hz, t), 2.85 (2H, d br, J-11.4 Hz, NCH_eq), 3.46 (2H, s, CH₂Ph), 7.20 - 7.35 (5H, m). b) 4-[4-(N-Benzoyl-N-methylamino)butyl]-l-benzylpiperidine (XVII)

To a stirred solution of (XVI) (1.30 g, 5.0 mmol) in acetone (7 mL) was added 1 N NaOH (5.5 mmol) then a solution of benzoyl chloride (0.61 mL, 5.25 mmol) in acetone (3 mL) was added dropwise. The reaction mixture was stirred at room temperature for 2 h and then poured in brine (20 mL), alkalinized with 1 N NaOH and extracted with diethyl ether (3 x 20 mL). The combined organic phases were dried over Na₂Sθ4 and concentrated in vacuo. The residue was purified by flash column chromatography eluting with CHCl3-MeOH (98:2, v/v) to afford 1.50 g of (XVII) as a light yellow oil. Yield.82%. IR (neat) cm-l ; 2900, 2840, 1620 (CO). iH-NMR (CDCI3) δ (ppm): 1.10 - 2.00 (13H, m), 2.85 - 3.10 (5H, m), 3.20 (IH, t br, J=7.0 Hz), 3.48 (2H, s, CH₂Ph), 3.51 (IH, m), 7.20 - 7.35 (5H, m), 7.37 (5H, s). c) 4-[4-(N-Benzoyl-N-methylamino)butyl]piperidine (XVIII)

0.74 g of 4-[4-(N-benzoyl-N-methylamino)butyl]piperidine (XVIII) was prepared from 1.08 g of 4-[4-(N-benzoyl-N-methylamino)butyl]-l- benzylpiperidine (XVII) as described in Example 1 point f). Yield 91%. IR (neat) cm"¹ : 3260 (NH), 2890, 2830, 1615 (CO). iH-NMR (CDCI3) δ: 0.90 - 1.75 (11H, m), 2.07 (IH, s br, NH), 2.56 (2H, t br, J=10.0 Hz, NCH_ax), 2.86 - 3.10 (5H, m, NCH_eq and NCH3), 3.20 (IH, t br, J=5.7 Hz), 3.50 (IH, t br, J=5.7 Hz), 7.35 (5H, s). d) 4-[4-(N-Benzyl-N-methylamino)butyl]piperidine (XIX) 0.57 g of 4-[4-(N-benzyl-N-methylamino)butyl]piperidine (XIX) was prepared from 0.65 g of 4-[4-(N-benzoyl-N-methylamino)butyl]piperidine (XVIII) as described in Example 1 point d), heating at reflux for 1 h. Yield 92%.

IR (neat) cm"¹: 2910, 2840, 2780, 1450. iH-NMR (CDCI3) δ (ppm): 0.90 - 1.80 (12H, m), 2.17 (3H, s), 2.34 (2H, t, J=6.4 Hz), 2.56 (2H, td, .7=11.1, 2.8 Hz, NCH_ax), 3.03 (2H, dt, .7=11.1, 2.8 Hz, NCH_eq),

3.45 (2H, s, CH₂Ph), 7.20 - 7.35 (5H, m). e) 5-{[4-[4-(N-Benzyl-N-methylamino)butyl]-l-piperidinyl}acetyl-5H- dibenz [b,fj azepine

0.92 g of 5-{[4-[4-(N-benzyl-N-methylamino)butyl]-l-piperidinyl}acetyl-5H- dibenz[b,f]azepine was prepared from 0.53 g of 5-(chloroacetyl)-5H- dibenzfb_j jazepine and 4-[4-(N-benzyl-N-methylamino)butyl]piperidine (XLX) as described in Example 1 point g). Yield 95%.

Elemental Analysis C H N

(theor. %) 80.28 7.96 8.51 (found %) 80.07 8.00 8.49

IR (neat) cm"¹: 3020, 2920, 1670.

*H-NMR (CDCI3) δ (ppm): 1.00 - 2.00 (13H, m, CH₂, CH and NCH_ax), 2.15

(3H, s, NCH3), 2.32 (2H, t, J=6.8 Hz, CH^NMeBn), 2.66 (2H, d, J=9.1 Hz, NCH_eq), 2.76 (IH, d, .7=14.3 Hz, COCHH'), 3.07 (IH, d, J=14.3 Hz, COCHH'), 3.44 (2H, s, NCH₂Ph), 6.92 (2H, dd, .7=15.7, 11.8 Hz, CHOH), 7.20 - 7.45 (8H, m) EXAMPLE 3 5-{[4-[4-(Diethylamino)ethyl]-l-piperidinyI}acetyl-5H-dibenz[A_j ]azepine

0.30 g of 5-{[4-[4-(N-benzyl-N-methylamino)butyl]-l-piperidinyl}acetyl-5H- dibenz[b,fjazepine was prepared from 0.25 g of 5-(chloroacetyl)-5H- dibenz[b_j ]azepine and 4-[4-(diethylamino)ethyl]piperidine as described in Example 1 point g). Yield 77%. Elemental Analysis C H N

(theor. %) 77.66 8.45 10.06

(found %) 77.50 8.50 10.00

^ΪH-NMR (CDCI3) δ (ppm): 1.00 (6H, t, 7=6.8 Hz, N(CH₂CH )₂), 1.05 - 2.05

(9H, m, CH₂, CH and NCH_ax), 2.39 (2H, m, CH₂NEt₂), 2.49 (4H, q, J=6.8 Hz,NCH₂CH₃), 2.65 (2H, d, J=9.0 Hz, NCH_eq), 2.78 (IH, d, .7=14.7 Hz, COCHH'), 3.07 (IH, d, .7=14.7 Hz, COCHH), 6.92 (2H, dd, .7=13.5, 11.1 Hz, CHOH), 7.30 - 7.45 (8H, m) EXAMPLE 4 a) N,N-DiethyI-6-bromohexanamide (XX) A solution of 6-bromohexanoyl chloride (2.0 g, 9.37 mmol) in acetone (4 mL) was added dropwise to a solution of diethylamine (4.9 mL, 46.85 mmol) in water (8 mL) at 0°C. The mixture was stirred for 2h at 0°C, then poured in brine (40 mL) and extracted with diethyl ether (3 x 30 mL). The combined extracts were washed with water, dried over Na2Sθ4 and concentrated in vacuo to give 2.2 g of (XX) as a yellow oil. Yield 94%.

IR (neat) cm"¹ : 2920, 1610 (CO), 1420. iH-NMR (CDCI3) δ (ppm): 1.09 (3H, t, J=7.0 Hz, CONCH₂CH₃) 1.14 (3H, t,

.7=7.0 Hz, CONCH'₂CH'₃), 1.35 - 1.75 (4H, m), 1.87 (2H, m), 2.29 (2H, t, J=7.8 Hz), 3.20 - 3.46 (6H, m). b) N,N-Diethyl-7-(4-pyridyl)heptanamide (XXI)

A 50% suspension sodium amide in toluene (0.75 g, 9.60 mmol) was added to a solution of 4-picoline (0.93 mL, 9.60 mmol) in anhydrous toluene (10 mL) under nitrogen atmosphere. The mixture was stirred for 30 min at 80°C, cooled to room temperature then amide (XX) (1.6 g, 6.40 mmol) in toluene (6 mL), was added. The resulting solution was stirred at 80°C for 6 h, then poured in brine (30 mL) and the organic layer was separated. The water layer was extracted with diethyl ether (3 x 20 mL). The combined extracts were washed with water, dried over Na₂Sθ4 ^and concentrated. The residue was purified by flash chromatography column with petroleum ether-acetone (65:35, v/v) to afford 0.51 g of (XXI) as a yellow oil. Yield 30%. iH-NMR (CDC1₃) δ (ppm): 1.10 (3H, t, J=7.0 Hz), 1.15 (3H, t, J=7.0 Hz), 1.27 - 1.80 (8H, m), 2.26 (2H, t, J=7.0 Hz, CH₂CONEt₂), 2.58 (2H, t, J=7.0 Hz, CH₂C5H₄N), 3.32 (4H, m,), 7.08 (2H, dd, .7=4.1, 1.3 Hz), 8.45 (2H, dd, .7=4.1, 1.6 Hz). c) N,N-Diethyl-7-(4-piperidinyl)heptanamide (XXII)

Pyridyl derivative (XXI) (0.47g) was dissolved in 1.9 M in methanol HCl (20 mL) and stirred at room temperature for 40 min. The solvent was evaporated, the residue was dissolved in acetic acid and Pt0₂ (25 mg) was added. The mixture was maintained under a hydrogen atmosphere at room temperature and atmospheric pressure for 24 h. After that, the catalyst was filtered off and the filtrate was basified with 32% NaOH then extracted with diethyl ether. The combined extracts were washed with water, dried over Na₂Sθ4 and concentrated to afford 0.41 g of (XXII). Yield 85%. iH-NMR (CDCI3) δ (ppm): 1.09 (3H, t, J=7.0 Hz), 1.15 (3H, t, J=7.0 Hz), 1.20 - 1.80 (16H, m), 2.27 (2H, t, J=7.8 Hz, CH₂CONEt₂), 2.55 (2H, td, .7=11.4, 2.2 Hz, NCH_ax), 3.02 (2H, dt, .7=11.4, 2.1 Hz, NCH_eq), 3.32 (4H, m, NCH₂CH₃) d) 4-[4-(Diethylamino)heptyl]piperidine (XXIII)

0.32 g of 4-[4-(diethylamino)heptyl]piperidine (XXIII) was prepared from

0.35 g of N,N-diethyl-7-(4-piperidinyl)heptanamide (XXII) as described in

Example 1 point d), heating under reflux for 2 h. Yield 96%. iH-NMR (CDCI3) δ (ppm): 1.01 (6H, t, J=6.9 Hz), 1.10 - 1.80 (18H, m), 2.39

(2H, t, 7=7.7 Hz, CH₂NEt₂), 2.51 (4H, q, 7=6.9 Hz, NCH₂CH3), 2.55 (2H, m,

NCHax), 3.03 (2H, dt, 7=11.8, 2.8 Hz, NCH_eq). e) 5-{ [4-[4-(Diethylamino)heptyl]-l-piperidinyl}acetyl-5H-dibenz[Λ^]azepine 0.32 g of 5-{[4-[4-(diethylamino)heptyl]-l-piperidinyl}acetyl-5H- dibenz[b,fjazepine was prepared from 0.29 g of 5-(chloroacetyl)-5H- dibenz[bj/]azepine and 4-[4-(diethylamino)heptyl]piperidine (XXIII) as described in Example 1 point g), heating to reflux for 2 h. Yield 61%.

Elemental Analysis C H N

(theor. %) 78.80- 9.30 8.62 (found %) 78.75 9.31 8.59

IR(neat) cm-l; 3015,2900, 1665.

!H-NMR (CDCI3) δ (ppm): 1.00 (6H, t, 7=7.1 Hz, N(CH CH.3)₂), 1.05 - 2.05

(19H, m, CH₂, CH and NCH_ax), 2.37 (2H, m, , CH₂NEt ), 2.49 (4H, q, 7=7.1 Hz,

NCH₂CH₃), 2.65 (2H, d, 7=9.7 Hz, NCH_eq), 2.78 (IH, d, 7=15.1 Hz, COCHH), 3.07 (IH, d, 7=15.1 Hz, COCHH), 6.92 (2H, dd, 7=12.5, 11.0 Hz, CHOH), 7.30

- 7.45 (8H, m)

EXAMPLE 5

5-{[2-[(Diethylamino)methyl]-l-piperidinyl}acetyl-5H-dibenz[A^lazepine 0.26 g of 5-{[2-[(diethylamino)methyl]-l-piperidinyl}acetyl-5H- dibenz[b_j ]azepine was prepared from 0.20 g of 5-(chloroacetyl)-5H- dibenz[b,/]azepine and 2-[(diethylamino)methyl]piperidine as described in

Example 1 point g). Yield 87%.

IR (neat) cm-l : 3020, 29200, 1670. iH-NMR (CDCI3) δ (ppm): 0.95 (6H, t, 7=7.0 Hz, N(CH₂CH₃)2), 1.10 - 2.80 (15H, m), 2.98 (0.5H, d, COCH₂N), 3.27 (0.5H, d, COCH₂N), 3.46 (0.5H, d, COCH₂N), 3.72 (0.5H, d, COCH₂N), 6.95 (2H, s, CHOH), 7.25 - 7.45 (8H, m). EXAMPLE 6 a) 5-(Chloroacetyl)-10-methoxy-5H-dibenz[ , ]azepine

To a solution of 10-methoxy-5H-dibenz[b,fJazepine (2.61 g, 11.7 mmol) in tetrahydrofuran (40 mL) were added N,N-dimethylaniline (1.55 mL, 12.3 mmol) and chloroacetyl chloride (0.98 mL, 12.3 mmol) and the solution was refluxed for 2 h. The reaction was quenched with 200 mL of brine and extracted with diethyl ether (3 x 80 mL). The combined organic phases were washed with 0.1 N HCl (2 x 100 mL) to remove N,N-dimethylaniline, with 0.1 N NaOH (2 x 100 mL) to remove chloroacetylacetic acid then with water. The oOrganic layer was dried over Na₂S04, filtered and evaporated in vacuo to give 3.50 g of 5-(chloroacetyl)- 10-methoxy-5H-dibenz[b_j 3azepine as yellowish powder. Quantitative yield. IR (nujol) cm- 1 : 3040, 1670 (CO). iH-NMR (CDCI3) δ (ppm): 3.73 (0.5H, d, 7=5.1 Hz, COCHH'Cl), 3.80 (0.5H, d, 7=5.1 Hz, COCHH'Cl), 3.88 (3H, d, 7=4.5 Hz, OCH3), 3.97 (0.5H, d, 7=5.1 Hz, COCHH'Cl), 4.03 (0.5H, d, 7=5.1 Hz, COCHH'Cl), 6.15 (IH, d, 7=13.8 Hz, CH -OCH3), 7.26 - 7.55 (7H, m), 7.77 (IH, t br, 7=7.7 Hz). b) 5-{ [4-[4-(Diethylamino)butyl]-l-piperidinyI}acetyl-10-methoxy-5H- dibenz[A,/]azepine

4.8 g of 5-{ [4-[4-(diethylamino)butyl]-l-piperidinyl}acetyl-10-methoxy-5H- dibenz[b,fjazepine was prepared from 3.3 g of 5-(chloroacetyl)-10-methoxy-5H- dibenz[b_j ]azepine and 4-[4-(diethylamino)butyl]piperidine (XV) as described in Example 1 point g). Yield 92%.

Elemental Analysis C H N

(theor. %) 75.75 8.69 8.83

(found %) 75.61 8.70 8.78 IR (nujol) cm"¹ : 3065, 2929, 1674 (CO). iH-NMR (CDC1₃) δ (ppm): 1.00 (6H, t, 7=7.0 Hz, N(CH₂CH₃) ), 1.08 - 2.03 (13H, m, CH , CH and NCH_ax), 2.39 (2H, t, 7=7.1 Hz, CH₂NEt₂), 2.52 (4H, q, 7=7.0 Hz, NCH₂CH₃), 2.60 - 3.15 (4H, m, COCH₂N and NCHeq), 3.86 (3H, d, 7=3.8 Hz, OCH3), 6.11 (IH, d, 7=7.3 Hz, CHOOCH3), 7.20 - 7.48 (7H, m), 7.72 (IH, m) EXAMPLE 7 a) 5-(Chloroacetyl)-10-ethoxy-5H-dibenz[Λ, ]azepine

3.3 g of 5-(chloroacetyl)-10-ethoxy-5H-dibenz[b_j/]azepine was prepared from 2.55 g of 10-ethoxy-5H-dibenz[b_j ]azepine as described in Example 6 point a). Yield 98%.

^-NMR (CDCI3) δ (ppm): 1.48 (3H, m, OCH₂CH₃), 3.73 (0.5H, d, 7=6.1 Hz, COCHH'Cl), 3.80 (0.5H, d, 7=6.1 Hz, COCHH'Cl), 3.94 - 4.17 (3H, m, CH₂CH and COCHH'Cl), 6.14 (IH, d, 7=11.5 Hz, CHO-OEt), 7.26 - 7.55 (7H, m), 7.79 (lH, t br, 7=7.3 Hz). b) 5-{[4-[4-(Diethylamino)butyl]-l-piperidinyl}acetyl-10-ethoxy-5H- dibenz[Z>,/]azepine

0.21 g of 5- {[4-[4-(diethylamino)butyι]-l-piperidinyl} acetyl- 10-ethoxy-5H- dibenz[b,/]azepine was prepared from 3.3 g of 5-(chloroacetyl)-10-ethoxy-5H- dibenz[b_j ]azepine and 4-[4-(diethylamino)butyl]piperidine (XV) as described in Example 1 point g). Yield 92%.

Elemental Analysis C H N

(theor. %) 76.03 8.85 8.58

(found %) 75.88 8.92 8.60 IR (nujol) cm-¹ : 3065, 2929, 1674 (CO). iH-NMR (CDCI3) δ (ppm): 1.00 (6H, t, 7=6.9 Hz, N(CH₂CH₃)2), 1.08 - 2.05 (16H, m, CH₂, CH, OCH₂CH₃ and NCH_ax), 2.37 (2H, t, 7=7.1 Hz, CH NEt₂), 2.49 (4H, q, 7=6.9 Hz, N(CH CH₃)₂), 2.60 - 3.15 (4H, m, COCH₂N and NCHgq), 4.05 (2H, q br, OCH₂CH₃), 6.11 (IH, d, 7=5.1 Hz, CHOOEt), 7.20 - 7.46 (7H, m), 7.75 (lH, m). EXAMPLE 8 c) 5-(Chloroacetyl)-10-butoxy-5H-dibenz[A^]azepine 0.38 g of 5-(chloroacetyl)-10-butoxy-5H-dibenz[b_j ]azepine was prepared from

0.31 g of 10-butoxy-5H-dibenz[b_j/]azepme as described in Example 6 point a). Yield 94%. iH-NMR (CDC1 ) δ (ppm): 1.02 (3H, t, 7=6.8 Hz, 0(CH₂)₃CH₃), 1.48 - 1.67 (2H, m, 0(CH₂)₂CH₂CH₃), 1.74 - 1.95 (2H, m, OCH₂CH₂CH₂CH₃), 3.73 (0.5H, d br, 7=6.11 Hz, COCHH'Cl), 3.89 (0.5H, d br, 7=6.11 Hz, COCHH'Cl), 3.90 - 4.09 (3H, m, OCH₂(CH₂)2CH₃ and COCHH'Cl), 7.26 - 7.55 (7H, m), 7.77 (IH, m). d) 5- { [4- [4-(Diethylamino)butyl] -1 -piperidinyl } acety 1-10-butoxy-5H- dibenz[Z>,/| azepine 0.41 g of 5- {[4-[4-(diethylamino)butyl]-l -piperidinyl} acetyl- 10-butoxy-5H- dibenz[b, Jazepine was prepared from 0.38 g of 5-(chloroacetyl)-10-butoxy-5H- dibenz[b ]azepine and 4-[4-(diethylamino)butyl]piperidine (XV) as described in Example 1 point g). Yield 71%.

Elemental Analysis C H N

(theor. %) 76.55 9.15 8.12

(found %) 76.35 9.20 8.10

IR (nujol) cm-l : 3065, 2929, 1674 (CO). iH-NMR (CDCI3) δ (ppm): 1.00 (9H, m, N(CH₂CH₃)₂ and 0(CH₂)₃CH₃), 1.05 - 2.05 (17H, m, CH₂, CH, OCH₂(CH₂) CH₃ and NCH_ax), 2.37 (2H, t, 7=6.9 Hz, CH₂NEt₂), 2.49 (4H, q, 7=6.6 Hz, N(CH₂CH₃)₂), 2.60 - 3.15 (4H, m, COCH₂N and NCH_eq), 4.00 (2H, s br, OCH₂(CH₂)₂CH₃), 6.11 (IH, m, CHOOBu), 7.15 - 7.48 (7H, m), 7.72 (lH, m) EXAMPLE 9 a) 10-Phenoxy-5H-dibenz[£, ]azepine (VI)

A solution of phenol (0.43 g, 4.5 mmol) in 8 mL of anhydrous dimethyl sulfoxide was added with potassium t-butoxide (1.02 g, 4.5 mmol). The solution was stirred at room temperature for 30 min, then 5-acetyl-10-bromo-5H-dibenz[b,/]azepine (VII) (0.95 g, 3.0 mmol) was added and the mixture was stirred at 90°C for 20h. After cooling to room temperature, the reaction mixture was poured into water (100 mL) and extracted with diethyl ether (4 x 25 mL). The combined organic phases were washed with 2 N NaOH, dried over Na₂Sθ4 and concentrated to give 0.17 g of 10-phenoxy-5H-dibenz[b_j ]azepine (VI) as a yellow oil. Yield 20%. H-NMR (CDCI3) δ (ppm): 5.17 (IH, s br, NH), 6.23 (IH, s br, CHO-Ph), 6.55 -

7.42 (13H, m). b) 5-(Chloroacetyl)-10-phenoxy-5H-dibenz[^₅/]azepine

0.11 g of 5-(chloroacetyl)-10-phenoxy 5H-dibenz[b/]azepine was prepared from 0.13 g of 10-phenoxy-5H-dibenz[b jazepine (VI) as described in Example 6 point a). Yield 65%. iH-NMR (CDCI3) δ (ppm): 3.82 (IH, d, 7=13.3 Hz, COCHH'Cl), 4.02 (IH, d, 7=13.3 Hz, COCHH'Cl), 6.58 (IH, s, CHO-OPh), 6.95 - 7.55 (12H, m), 7.77 (lH, dd, 7=22.2, 7.5 Hz). c) 5-{[4-[4-(Diethylamino)butyl]-l-piperidinyl}acetyl-10-phenoxy-5H- dibenz [bj] azepine

0.11 g of 5-{[4-[4-(diethylamino)butyl]-l-piperidinyl}acetyl-10-phenoxy-5H- dibenz[b_j ] azepine was prepared from 0.11 g of 5-(chloroacetyl)-10-phenoxy-5H- dibenz[b_j ]azepine and 4-[4-(diethylamino)butyl]piperidine (XV) as described in Example 1 point g). Yield 67%.

Elemental Analysis C H N

(theor. %) 78.18 8.06 7.81

(found %) 77.99 8.10 7.75 iH-NMR (CDCI3) δ (ppm): 1.00 (6H, t, 7=6.8 Hz, N(CH CH₃)₂), 1.05 - 2.10 (13H, m, CH₂, CH and NCH_ax), 2.38 (2H, t, 7=6.9 Hz, CH₂NEt₂), 2.51 (4H, q, 7=6.8 Hz, N(CH₂CH₃)₂), 2.60 - 2.96 (3H, m, COCHH' and NCH_eq), 3.17 (IH, m, COCHH'), 6.54 (IH, d, 7=11.6 Hz, CHO-OPh), 6.95 — 7.49 (12H, m), 7.72 (IH, dd, 7=23.1, 7.4 Hz). EXAMPLE 10 a) 5-(6-Bromohexanoyl)-5H-dibenz[Λ₃7]azepine

0.30 g of 5-(6-bromohexanoyl)-5H-dibenz[b_j ]azepine was prepared from 0.16 g of 5H-dibenz[b_j ]azepine as described in Example 6 point a). Yield 96%. iH-NMR (CDCI3) δ (ppm): 1.20 - 1.81 (6H, m, -(CH₂)₃-), 1.92 (IH, m, COCHH'), 2.23 (IH, m, COCHH'), 3.32 (2H, t, 7=6.9, -CH₂-Br), 6.95 (2H, dd, 7=15.2, 12.2 Hz, CHOH), 7.25 - 7.50 (8H, m). b) 5-{ [4-[4-(Diethylamino)butyl]-l-piperidinyI}hexanoyl-5H- dibenz [bj] azepine 0.29 g of 5-{[4-[4-(diethylamino)butyl]-l-piperidinyl}hexanoyl-5H- dibenz[b Jazepine was prepared from 0.30 g of 5-(6-bromohexanoyl)-5H- dibenz[b/]azepine and 4-[4-(diethylamino)butyl]piperidine (XV) as described in Example 1 point g). Yield 71%.

Elemental Analysis C H N

(theor. %) 78.99 9.44 8.37

(found %) 78.80 9.55 8.25

IR (neat) cm"¹: 3024, 2929, 1679 (CO). iH-NMR (CDCI3) δ (ppm): 1.00 (6H, t, 7=6.8 Hz, N(CH₂CH₃)₂), 1.08 - 1.98 (20H, m, CH₂, CH, COCHH and NCH_ax), 2.20 (3H, m, COCHH' and CH₂N(CH₂CH₂)₂CH), 2.38 (2H, t, 7=6.9 Hz, CH₂NEt₂), 2.50 (4H, q, 7=6.8 Hz, N(CH₂CH3)₂), 2.82 (2H, d br, 7=10.2 Hz, NCH_eq), 6.92 (2H, dd, 7=15.8, 11.3 Hz, CHOH), 7.27 - 7.45 (8H, m). EXAMPLE 11 a) 5-(6-Bromodecanoyl)-5H-dibenz [bj] azepine

0.80 g of 5-(6-bromodecanoyl)-5H-dibenz[b_j/]azepine was prepared from 0.39 g of 5H-dibenz[b, ]azepine as described in Example 6 point a). Yield 94%. H-NMR (CDCI3) δ (ppm): 1.07 - 2.03 (15H,m, -(CH₂)₇- and COCHH), 2.20

(IH, m, COCHH^*), 3.38 (2H, t, 7=6.8 Hz, -CH₂-Br), 6.95 (2H, dd, 7=16.3, 13.0 Hz), 7.30 - 7.55 (m, 8H) b) 5-{ [4-[4-(Diethylamino)butyl]-l-piperidinyI}decanoyI-5H- dibenz [bj azepine 0.36 g of 5- {[4-[4-(diethylamino)butyl]-l -piperidinyl} decanoyl-5H- dibenz[b_j ]azepine was prepared from 0.80 g of 5-(6-bromodecanoyl)-5H- dibenz[b )azepine and 4-[4-(diethylamino)butyl]piperidine (XV) as described in Example 1 point g). Yield 34%. Elemental Analysis C H N (theor. %) 79.66 9.94 7.53

(found %) 79.45 9.90 7.50 iH-NMR (CDCI3) δ (ppm): 1.00 (6H, t, 7=7.0 Hz, N(CH₂CH₃) ), 1.07 - 2.00 (28H, m, CH₂, CH, COCHH' and NCH_ax), 2.20 (3H, m, COCHH^* and CH₂N(CH₂CH₂)₂CH), 2.40 (2H, t, 7=7.0 Hz, CH₂NEt₂), 2.51 (4H, q, 7=7.0 Hz, N(CH₂CH₃)₂), 2.88 (2H, d br, 7=10.6 Hz, NCH_eq), 6.92 (2H, dd, 7=16.5, 11.3 Hz, CHOH), 7.27 - 7.45 (8H, m) EXAMPLE 12

5- { [2- [(Diethylamino)methyl] -1 -piperidinyl } acetyl-10,11 -dihydro-5H- dibenz[Z>_j/] azepine

0.31 g of 5 - { [2- [(diethylamino)methyl] - 1 -piperidinyl } acetyl- 10,11 -dihydro- 5H-dibenz[b_j/]azepine was prepared from 0.22 g of 5-(chloroacetyl)-10,l l- dihydro-5H-dibenz[b |azepine and 2-[(diethylamino)methyl]piperidine as described in Example 1 point g). Yield 94%. Elemental Analysis C H N

(theor. %) 77.00 8.70 10.36

(found %) 75.60 8.62 10.00

IR (neat) cm"l : 3020, 2920, 1670 (CO). iH-NMR (CDC1₃) δ (ppm): 0.92 (6H, m, N(CH₂CH₃)₂), 1.10-3.95 (21H, m),

7.05-7.45 (8H, m).

EXAMPLE 13

5-{ [4-[4-(Diethylamino)butyl]-l-piperidinyl}acetyl-10,ll-dihydro-5H- dibenz[b | azepine 4.0 g of 5-{[4-[4-(diethylamino)butyl]-l-piperidinyl}acetyl-10,l l-dihydro-

5H-dibenz[b, ] azepine was prepared from 2.9 g of 5-(chloroacetyl)- 10,11 -dihydro-

5H-dibenz[b_j ]azepine and 4-[4-(diethylamino)butyl]piperidine (XV) as described in Example 1 point g). Yield 84%. Elemental Analysis C H N

(theor. %) 77.81 9.23 9.39

(found %) 77.58 9.17 9.42

IR (neat) cm"¹ : 3028, 2928, 1673 (CO). iH-NMR (CDCI3) δ (ppm): 1.00 (6H, m, N(CH₂CH.3)₂), 1.05 - 2.15 (13H, m, CH₂, CH and NCH_ax), 2.37 (2H, m, CH₂NEt₂), 2.50 (4H, q, 7=7.1 Hz,

N(CH₂CH₃)₂), 2.60 - 3.55 (8H, m, COCH₂N, CH₂-CH₂ and NCH_eq), 7.10 -

7.45 (8H, m).

PHARMACOLOGICAL EXPERIMENTATION

RECEPTOR BINDING ASSAY The binding affinity and the selectivity for muscarinic receptors of the compounds of the examples have been evaluated by using cloned human receptors.

Membranes of Hamster ovarian cells (Receptor Biology, Beltsville, MD, USA) expressing human Mi , M₂, M3 or M4 receptors were suspended in a 10 mM Tris- HCl solution at pH 7.2 containing 2 mM EDTA (plus 10% sucrose in the case of Mi , M3 and M4 receptors). Aliquots of the membrane fractions (0.02 mL) were incubated in duplicate with 10 increasing concentrations (from 10" 10 to 10"5 M) of the reference (4-DAMP) or test compounds. The radioligand [3H]N-methylscopolamine ([3H]NMS) was added to the membrane preparations at the final concentration of 0.5 nM for Mi , M₂, M4 receptors and 0.1 nM for M3 receptors. The membrane preparations were incubated for 60 min at 25°C or 27°C. Atropine (2.5 μM for M\ receptors, 1 μM for M2 receptors, 0.5 μM for M3 and M4 receptors) was used to determine the non-specific binding. The assay was terminated by rapid filtration under suction through Whatman GF/B glass fiber filters using a Tomtec 96 well harvester. After 4-5 washes with ice-cold buffer, the filters were put in plastic bags with 25 mL of Betaplate Scint® (Wallac) scintillant cocktail and the plastic bags were sealed. The 3H-radioactivity was counted using a Wallac 1205 Betaplate® liquid scintillation counter. Inhibition curves were analyzed with the software EBDA (McPherson, Elsevier Biosoft, Cambridge, U.K.) to determine the affinity constants (Kj) of the compounds for muscarinic receptors subtypes.

The results, expressed as Kj values and selectivity ratios for M2 muscarinic receptors over M\, M3 and M4 muscarinic receptors, are presented in Table 1. AF-DX 116, was used as reference compound.

Table 1

EXAMPLE Binding ; Affinity, Ki (nM) Selectivity Ratio 1 hMi hM₂ hM₃ hM₄ Mj/M₂ M₃/M₂ M4/M2

1 310 2.6 290 100 120 110 38

1020 55 1100 550 18 20 10

3 nd 620 nd nd - - -

4 nd 90 nd nd - - -

5 nd 110 nd nd - - -

6 360 20 420 100 18 21 5

7 1600 34 560 400 47 16 12

8 nd 120 nd nd - - -

9 nd 120 nd nd - - -

10 13 9.4 52 8.8 1.4 5.5 - 0.9

11 18 14 280 49 1.3 20 3.5

12 nd 150 nd nd - - -

13 810 12 110 370 67 92 31

AF-DX 116 2900 71 7700 1200 41 108 17 nd: not determined The compounds of the Examples 1, 6, 7, 10 and 13 showed a higher affinity than

AF-DX 116 for M₂ receptors. Compared to AF-DX 116 the compounds of the

Examples 1 and 13 exhibit a better selectivity profile.

IN VITRO FUNCTIONAL ACTIVITY

Guinea-Pig Atrium. Isolated left atria were prepared from male or female Duncan Hartley derived guinea pigs weighing 325 ± 25 g. Animals were sacrificed by C0₂ overexposure. Each atria was placed under 1 g tension in a 10 ml bath containing McEwen's solution at pH 7.4, bubbled with 95% 0₂/5% C0₂ at 32 °C and subjected to field stimulation by 70% maximum voltage, 2.5 Hz at 0.5 msec pulse width. The left atria was connected to an isometric transducer and two pen recorders and allowed to equilibrate for 60 minutes before initiating the field stimulation. Each tissue was accepted for experimental use only if 1 g or more of tension was obtained. Cumulative relaxation-response curve to methacholine was then generated with consecutive applications of 9 concentrations in 3 -fold increments ranging from 1 nM to 10 μM at 2 minutes intervals for a total of 18 minutes to establish the maximal response. In 4 separate tissues, similar methacholine concentration-response were carried out in the presence of test compound concentration (low, middle and high), following a 15 minutes incubation period. pA₂ values for test compounds were calculated by linear regression of the corresponding Schild plots. Table 2 reports the pA₂ values for the compounds of the Example 1, 6, 13.

Guinea-Pig Ileum. Segments (2-3 cm) of proximal ileum were suspended in 30 mL Tyrode solution at 37 ± 1 °C. The preparation was connected to an isometric strain gauge and was kept at a resting tension of 1 g. The changes in tension were registered through a polygraphic recorder. The ileum was stimulated with a sub-maximal concentration of acetylcholine (0.03 μM) every 4-5 minutes. The drugs were prepared in physiological saline solution. The activity of test compounds under investigation was expressed as a percent inhibition of the contraction induced by acetylcholine. The IC50 was calculated by non-linear regression. Table 2 reports the IC50 values for the compounds of the Example 1, 6, 13.

Table 2

EXAMPLE pA₂ IC50 (μM)

1 7.08 0.54

6 6.20 6.79

13 5.61 1.19

These functional in vitro studies confirm that compounds of the Example 1 and 6 have a better affinity for M₂ muscarinic receptors than for M3 muscarinic receptors.

ANTIBRADYCARDIC ACTIVITY IN RATS.

Antibradycardic activity of the Example 1 was evaluated in vivo in comparison to AF-DX 116. The effects of these compounds on acetylcholine- induced bradycardia in rats were evaluated after intravenous and intraduodenal administration.

Intravenous studies. Male Sprague-Dawley rats weighing 270 ± 5 g were housed for a minimum of 5 days before the experiments under a 12-hour light- dark cycle at a room temperature (20 ± 2 °C) and 55% minimum humidity. Rats had free access to commercial chow and tap water. Animals were anaesthetized with urethane (1.5 g/kg, i.p.) and their body temperature was maintained at 37°C with a heating pad. A 2-cm incision was made in the middle of the neck and a tracheotomy tube was inserted. The carotid aorta was isolated and connected with a catheter to a pressure transducer. Heart rate and blood pressure were recorded and analyzed with the software HEM (Notocord, Croissy sur Seine, France). Another catheter was introduced in the left jugular vein for acetylcholine perfusion. A third catheter was introduced in the right jugular vein for vehicle, test or reference compound administration. After a 10-min stabilization period, acetylcholine (50 μg/min/kg) was continuously infused intravenously until stabilization of the blood pressure. Vehicle (sterile saline, 1 mL/kg) was administered as an intravenous bolus. Test compounds were then injected as boluses at increasing concentration (10, 50, 250 μg/kg). Finally, atropine (10 μg/kg) was injected intravenously. At the end of experiments, rats were killed by urethane overdose. Four animals were used for each dose of the test compounds. ED50 values were estimated by non-linear regression and expressed as mean ± SEM. Table 3 reports the ED50 values of the compounds of the Example 1 and AF-DX 116 for systolic blood pressure and heart rate.

Table 3

COMPOUND ED₅₀ (μg/Kg)

Systolic Blood Heart Rate

Pressure

EXAMPLE 1 80 ± 1 55 ± 4

AF-DX 116 56 ± 1 13 ± 2

This study demonstrated that the Example 1 , when administered intravenously in the rat, is able to antagonize an acetylcholine action mediated by M2 muscarinic receptor subtypes. The effect of this compound was not significantly different

Intraduodenal studies. Male Sprague-Dawley rats, weighing 315 ± 3 g were housed, two per cage, under a 12-hour light-dark cycle at a room temperature (20 ± 2 °C) and 55% minimum humidity, for a minimum of 5 days before the experiments. Animals had free access to commercial rat chow and tap water. Rats were anaesthetized with urethane (1.5 g/kg, i.p.) and body temperature was maintained at 37°C with a heating pad. A two-cm incision was made at the middle of the neck and a tracheotomy tube was inserted. A catheter, filled with sterile heparin solution (5 U/mL), was inserted in the carotid aorta and connected to a pressure transducer. Heart rate and blood pressure were recorded and analyzed with the software HEM (Notocord, Croissy sur Seine, France). After a 10-min stabilization period, a bolus of acetylcholine (30 μg/kg) was administered intravenously. Then, vehicle (sterile saline, 1 mL/kg) or test compounds (1, 5 or 15 mg/kg) were administered through a catheter inserted in the duodenum. Forty- five minutes later, a second bolus of acetylcholine was administered intravenously. At the end of experiments, rats were killed by urethane overdose. Ten animals were used for each dose of test compounds. Five rats were employed for the vehicle administration. Statistical comparisons were carried out using one- way analysis of variance for completed randomized block. The test compounds were administered as maleate salts.

Following acetylcholine administration, heart rate decreases of 225 ± 13 beats/min corresponding to a reduction of 60% of the control value. Figure 1 represents the effects of the Example 1 and AF-DX 116 on the acetylcholine-induced bradycardia in the rat.

The compound of the Example 1 was able to counteract the acetylcholine-induced bradycardia in a dose — dependent fashion. At the dose of 15 mg/kg, this compound appeared more effective then AF-DX 116. HEMODYNAMIC EFFECTS IN CONSCIOUS DOGS.

The effects of oral administration of the Example 1 on nocturnal bradycardia were investigated in conscious freely moving dogs. The effects of this compound on cardiac conduction times as well as on systolic and diastolic blood pressure were also evaluated. Experiments were performed on 8 beagle dogs, 4 males and 4 females, weighing between 11 kg and 13 kg.

Hemodynamic variables and electrocardiograms were recorded using a surgically implanted telemetric transmitter (model TL2M10-D70, Data Science Inc., St. Paul, Minnesota) in freely moving animals. Implantation was performed at least 10 days before the drug administrations under general anaesthesia (20 mg/kg i.v. thiopental followed by 1-1.5% i.v. halothane). The telemetric transmitter was implanted in the left flank and the sensor catheter was introduced into the femoral artery of the animals. The ECG leads of transmitters were placed in lead II, with one electrode on the right forelimb and one electrode on the left hindlimb. Measurements were done in the animal room. Data were transmitted to an on-line-data acquisition system by means of a RLA2000 radio receivers (Data Science Inc., St. Paul, Minnesota). A sampling rate of 500 Hz and an ART 1.0 telemetry data acquisition system (Data Science Inc., St. Paul, Minnesota) were used. Heart rate, systolic and diastolic blood pressure were measured for 15 seconds every 5 minutes and averaged over 1-hour periods. Electrocardiogram (lead II) was recorded for 15 seconds every 5 minutes and values of conduction times were determined at each time point. Measurements began at least one hour before drug administration and lasted for 12 (for QT and PR and QRS intervals) or 24 (for heart rate, systolic and diastolic blood pressure) hours after dosing. Any gross behavioural or autonomic changes, observed during the experiment, were recorded.

Placebo and the Example 1 (2, 4 and 8 mg/kg) were orally administered according to a randomised four-way, crossover design with a washout period of at least one week. The drug was administered as a maleate salt and formulated in gelatine capsules. Administrations were done approximately at 10:00 p.m. by oral gavage.

Figure 2 shows the time course of heart rate for placebo and 3 doses of the Example 1 during the 24 hours post-treatment.

After placebo administration, a consistent reduction over time in heart rate was observed during the night-time period (-15%). The Example 1 antagonised the nocturnal bradycardia and shortened QT interval. The effect of the drug reached statistically significance, compared to placebo, with the highest dose of 8 mg/kg (+19%» on heart rate and -4% on QT interval). The effect on heart rate lasted for the entire 24-hour observation period. Nocturnal systolic and diastolic blood pressure were not significantly affected by the Example 1. No other signs of peripheral or central cholinergic block were observed at any dose. The results of this study demonstrated that oral administration of the Example 1 produces long- lasting hemodynamic effects in the conscious dogs. IN VrVO FUNCTIONAL SELECTIVITY.

Potential effects caused by antagonism of the other muscarinic receptor subtypes (mainly M3) were investigated. Gastric Emptying in Rats. Gastric emptying was evaluated by measuring the rate of disappearance on an aqueous suspension of phenol red from the stomach. Example 1 and AF-DX 116 were examined at the dose of 15, 50 and 150 mg/kg by the oral route and administered to groups of 8 animals. Atropine (20 mg/kg) was used as positive control drug. Wistar rats (200 - 300 g) were used in this study. Animals were housed in cages of standard dimension with sawdust. The animal house was kept at a temperature of 19 - 23°C and a relative humidity of 45 - 65% with non-recycled filtered air changed approximately 10 times per hour. A 12-hour day-night cycle was adopted with light on at 7:30 a.m. Rats received commercial food. Tap water was available ad libitum in polycarbonate feeder bottles with a stainless steel nipple. The day prior the study, animals were kept on a water only fast in cages with grid floors in order to minimize coprophagia. On the study day, animals were dosed as defined by the randomization plan. Sixty minutes after test compounds administration, animals were given orally 1.5 mL suspension of 0.05% phenol red in 2.5% carboxymethylcellulose (kept at 37°C ± 0.5). Ten minutes after the administration of phenol red, animals were sacrificed by pentobarbital overdosing (i.p.). Following midline laparotomy and ligature of the pylorus, the stomach was cut longitudinally and placed in 30 mL of a 0.9% NaCl solution. Twenty-four hours after, the stomach was removed and the volume adjusted to 36 ml with 0.9% NaCl and 4 mL of 1 M NaOH was added. After homogenization, the mixture was centrifuged at 3,000 rpm for 10 minutes. Phenol red concentrations were measured by spectrophotometry at 550 nM.

Intestinal Transit Time in Rats. Intestinal transit time was evaluated by measuring the distance covered in the intestines by a suspension of vegetal charcoal orally administered. Compound of the Example 1 and AF-DX 116 were examined at the dose of 15, 50 and 150 mg/kg by the oral route and administered to groups of 8 animals. Atropine (20 mg/kg p.o.) was used as positive control drug. Wistar rats (200 -300 g) were used in this study. Animals were housed in cages of standard dimension with sawdust. The animal house was kept at a temperature of 19 - 23°C and a relative humidity of 45 - 65% with non-recycled filtered air changed approximately 10 times per hour. A 12-hour day-night cycle was adopted (7.30 a.m. - 7.30 p.m.). Rats received commercial food. Tap water was available ad libitum in polycarbonate feeder bottles with a stainless steel nipple. The day prior the study, animals were kept on a water only fast in cages with grid floors in order to minimize coprophagia. On the study day, animals were dosed as defined by the randomization plan. Sixty minutes after test compounds administration, animals were given orally 2 mL suspension of 10% charcoal in 2.5% carboxymethylcellulose (kept at 37°C ± 0.5). Fifteen minutes after the carchoal administration, animals were sacrificed by cervical dislocation. Following midline laparotomy, the intestines were rapidly removed from the pylorus to the extremity of the cecum and spread out on a glass plate. The total length of intestine and the distance covered by the carchoal were then measured immediately afterwards.

Salivary Secretion in Rats. Salivary secretion was evaluated by measuring the weight gained by sublingual foam cubes after subcutaneous administration of oxotremorine. Compound of the Example 1 and AF-DX 116 were examined at the dose of 15, 50 and 150 mg/kg by the oral route and administered to groups of 8 animals. Atropine (10 mg/kg p.o.) was used as positive control drug. Male Sprague-Dawley rats (200 - 300 g) were randomly allocated to test treatments. Fifty minutes after test compounds or vehicle administration, rats were injected subcutaneously with oxotremorine (0.5 mg/kg). Ten minutes after oxotremorine administration, the mouth of the animals was opened with a gentle pressure on a snare allowing the placement in oral cavity of a pre-weighed, absorbent foam cube. Foam cubes were hold for 10 seconds and immediately after re-weighed. The difference between the first and second weight represents the saliva secreted. Pupil Diameter in Rats. Pupil diameter was evaluated under constant bright light with a dissecting microscope. Compound of the Example 1 and AF- DX 116 were examined at the dose of 15, 50 and 150 mg/kg by the oral route and administered to groups of 8 animals. Atropine (0.3 mg/kg p.o.) was used as positive control drug. Sprague-Dawley rats (200 - 250 g) were randomly allocated to test treatments. Pupil diameter was measured just before and 60 minutes after test compounds of vehicle administration.

In all in vivo functional selectivity studies, statistical comparisons were carried out using one-way analysis of variance for completed randomized block. Mean percent changes, compared to controls, of rate of gastric emptying, intestinal transit time, salivary secretion and pupil diameter after the different treatments are listed in Table 4.

Table 4

Gastric Intestinal Salivary Pupil

Emptying Transit Secretion Diameter

Atropine -84 ±10** -40 ±6** -54 ±7** 855 ±17**

AF-DX 116

15 mg/kg 0 + 8 6±5 -22 ±8 12 ±5

50 mg/kg -74+11** -25 ± 8* -18±9 176 ±67*

150 mg/kg -87 ±13** -36 ±5** -12 ±9 631 ±90**

Example 1

15 mg/kg 7±9 -2 ±5 2 ±10 49 ±33

50 mg/kg -48 ± 19** -19 ±8 -13 ±13 4±2

150 mg/kg -107 ±14** -31 ±9** -5 ±12 7±8

*p < 0.05 vs controls; **p < 0.01 vs controls

These studies show that in rats the antibradycardic effective doses of the Example 1 have virtual no effects on physiolgical functions mainly mediated by M3 and Mi cholinergic receptors. The compound appears to be more selective than AF- DX 116 especially as far as the pupil diameter is concerned where a favorable M1/M2 selectivity may play an important role. MULTIDIMENSIONAL SCREENING

A battery of 14 tests (_^multidimensional screening") was carried out for compound of the Example 1 at Huntingdon Life Sciences (Huntingdon, U.K.) to explore a wide range of potential activities of the compound in a series of standardized in vivo and vitro models.

The results obtained in each test of the multidimensional screening" are summarized in Table 5.

Table 5 Test Dose/Concentration Outcome

Irwin 30, 100, 300 mg/kg p.o. inactive

Anti-aggressive 100 mg/kg p.o. inactive

Anti-convulsivant 100 mg/kg p.o. inactive

Anti-depressant 30 mg/kg p.o. inactive

Analgesic (Writhing) 100 mg/kg p.o. inactive

Anti-Parkinson 300 mg/kg p.o. inactive

Anoretic 30 mg/kg p.o. inactive

Antihypertensive 100 mg/kg p.o. inactive

Anti-arrhytmic 100 mg/kg i.p. inactive

Pulmonary thromboembolism 100 mg/kg p.o. inactive

Diuretic 30 mg/kg p.o. inactive

Anti-inflammatory 100 mg/kg p.o. inactive

Platelet aggregation 100 μg/mL inactive

Immunostimulation/immunosuppression 20 mg/kg/day p.o. inactive

Bronchodilator 300 μg/mL inactive

The compound of the Example 1 was ineffective in all the activities explored, thus confirming a very selective pharmacological profile. In the Irwin test no abnormal signs were observed and no deaths occurred during the 7-day post-dose observation period at any of the doses tested. EFFECTS ON CORTICAL RELEASE OF ACETYLCHOLINE IN RATS An in vivo microdialysis study was carried out to examine the effects of the Example 1 on cortical release of acetylcholine in rats. A modified version of the microdialysis technique described by Cuadra et al. (7 Pharmacol Exper Ther 1994; 270: 277-284) was adopted.

Male Sprague-Dawley rats (250-330 g) were used. Rats were anesthetized with sodium pentobarbital (50 mg/kg i.p.) and mounted in a Kopf stereotaxic frame, with the incisor bar at 3.3 mm below intramural zero. Animals were implanted with a dialysis fiber inserted transversally in the cortex ( coordinates vs. bregma: A + 1.0 and V — 2.0). The probe was secured to the skull with screws and dental cement and the skin was sutured. Rats were placed into individual acrylic bowls and left to recover for at last 16 hours without perfusion. The experiment was commenced the following day. The microdialysis probe was connected to a 2-channel swivel and perfused with Ringer's solution (147 mM NaCl; 4.0 mM KC1; 4.0 mM CaCl₂) at the constant flow rate of 3 μL/min. Donepezil 3 μM was added to the Ringer's solution to prevent the hydrolysis of acetylcholine. The compound of the Example 1 was administered at time 0 after collecting baseline samples 90, 60 and 30 minutes prior to the administration. The compound was administered as maleate salt at the final concentrations of 3, 15 and 50 μM. It was dissolved in Ringer's solution and administered intracortically via the dialysis probe. Samples of the dialysate were collected into vials every 30 minutes until 330 minutes. Acetylcholine concentrations in the dialysate samples (40 μL) were measured by HPLC with electrochemical detection (ECD). The HPLC-ECD system consisted of a Coulochem II (ESA, Bedford, MA, USA) coupled with an ESA-580 pump. The mobile phase (100 mM Na₂HP04; 0.5 mM tetramethyammonium chloride; 0.005%) Reagent MB microbicide; 2.0 mM 1-octanesulfonic acid salt and 150 μM EDTA; pH 8.0) was filtered before passing trough a Prodigy ODS3 analytical column (C18 reverse phase, 5 μm, 150 x 3.2 mm, Phenomenex) at a flow rate of 0.600 mL/min. The column was coupled to an immobilized enzyme reactor containing acetylcholinesterase to convert acetylcholine to choline and acetic acid. The enzyme reactor also contained choline oxidase, which converts choline to betaine and hydrogen peroxide. Electrochemical detection of the latter was performed using an ECD with a platinum electrode set a +300 mV. The columns were maintained at the constant temperature of 35° C. The peaks of ACh in brain dialysates were displayed, integrated and stored by means of a Kontron Instrument 450. Quantification was made by comparing peak heights of the samples to a standard curve.

Figure 3 reports mean cortical acetylcholine concentrations after intracortical administration of the Example 1. The results of this study demonstrated that the compound of the Example 1, intracortical administered, was able to increase the acetylcholine concentrations in the rat cortex in a dose-response fashion. ANALGESIC ACTIVITY IN MICE.

The modified hot-plate procedure described by O'Callaghan and Holtzman was adopted {J Pharmacol Exper Ther 1975; 192: 497-505). Male Swiss albino mice (20 - 30 g) were used. Animals were kept at 22 ± 1 C° with a 12-hour light- dark cycle, with food and water ad libitum. Mice were placed on a stainless steel thermostat plate set at 50 ± 0.1 C°. A plastic cylinder was used to confine the mice to the heated surface of the hot-plate. The licking latency was defined as the time elapsing from thermal exposure and the licking of the fore or hind paws. A cut-off time of 75 seconds was adopted for licking latency in order to avoid unethical suffering and injury of animals. Mean baseline licking latency was determined with three measurements before test compounds administration. Test compounds were administered by subcutaneous route to groups of 12 - 17 animals. Compound of the Examples 1, 6, 13 and AF-DX 116 were evaluated at the doses of 5 mg/kg. Oxotremorine (0.2 mg/kg s.c.) was used as positive control drug. After test compounds administration, licking latencies were measured at 15-min intervals for 150 min. Statistical comparisons were carried out using repeated measures analysis of variance.

Figure 4 reported the licking latencies (mean ± SEM) after subcutaneous administration of oxotremorine (0.2 mg/kg), AF-DX 116 (5 mg/kg) and compounds of the Example 1, 6 and 13 (5 mg/kg) to groups of 12-17 mice. Compounds of the Example 1, 6, 13 and AF-DX 116 produced weak and transient effects on licking latencies suggesting a poor penetration of the blood brain barrier.

Claims

1. A compound of formula:

wherein X-Y represents CH₂-CH₂, CHOH, CH R3;

R\ represents hydrogen or linear or branched C1.4 alkyl;

R₂ represents linear or branched Cχ_4 alkyl, phenyl, benzyl or phenethyl;

R3 represents hydroxyl, linear or branched Cι _4 alkoxy, phenoxyl; m and n are independently an integer 1 to 10; and the salts thereof, obtained by addition of pharmacologically acceptable inorganic or organic acids.

2. Pharmaceutical composition containing an effective dose of a compound of formula (I) as claimed in claim 1 or of a salt thereof, in combination with pharmacologically acceptable diluents and excipients.

3. The use of the compounds of claim 1 for the preparation of medicaments for the treatment of cardiovascular, cognitive and neurodegenerative disorders.