Field of the Invention

The present invention is directed toward new sulfone ester analogues of iso-Clozapine [RN 1977-08-81 and their pharmaceutically acceptable salts, to processes for preparing such compounds, pharmaceutical preparations of such compounds and the use of such compounds in manufacture of a pharmaceutical preparation. Pharmaceutical preparations of these compounds are useful for central nervous system disorders, such as for a therapeutic effect on one or several of the following receptor systems: dopamine (DA; D1 - D4), 5-HT2A, 5-HT2C, α1 and muscarinic (M1 -M4) in mammals.

Background of the Invention

In psychotic/schizofrenic patients evidence indicates that the neurotransmission in the central nervous system (CNS) may be disturbed. These disturbances seem to involve the neurotransmitters dopamine (DA, D1 -D4 receptors), noradrenaline |NA, α1 receptors), acetylcholine (ACh, 1-M4 receptors) and 5-hydroxytryptamine (5-HT, 5-HT2A and 5- HT2C receptors). ¹ The drugs most frequently used in the treatment of psychoses are considered to act by altering the neurotransmission of one or several of these receptor systems. The mechanism of action for typical/classical anti-psychotic drugs used to treat psychosis is generally believed to be through blockage of DA D2 receptors. ^{2, 3} - ⁴ The atypical anti-psychotic drug Clozapine [RN 54241-01 -9] is an efficient anti-psychotic agent ⁵ and binds with high affinity to DA D1 and D4 receptors and with moderate affinity to DA D2 and D3 receptors. ⁶ In addition, Clozapine binds with high affinity to α1 receptors, muscarinic 1-M4 receptors and 5-HT2A and 5-HT2C receptors. ⁶ Clozapine has a low propensity to induce extrapyramidal side-effects in humans and catalepsy in rats. In addition, Clozapine does not increase the prolactine levels, something which the classical neuroleptics (e. g. haloperidol and chlorpromazine) potently do. However, the use of Clozapine is hampered by rare incidences of a serious side-effect (blood toxicity, aranulocytosis) . ^B Expensive, continuous blood monitoring makes the use of Clozapine tedious and expensive and the use of Clozapine is therefore limited to psychotic patients not responding to the first and second choice of drug therapy. ⁷

Several Clozapine analogues have been evaluated but, to date, no marketed compound with a Clozapine-like structure and Clozapine-like pharmacological and clinical profile, i. e.

an analogue that is also free from side-effects, has been found. ⁷

A very interesting and, for the present invention, a highly relevant fact is that the 2-CI isomer of Clozapine (iso-Clozapine, [RN 1977-08-81) has been classified as a typical neuroleptic agent. ⁸ We surprisingly found that the iso-Clozapine sulfone analogues of the present invention display atypical neuroleptic properties.

Information Disclosure Statement.

The following documents could be important in the examination of this application:

iso-Clozapine references: 9, 10, 1 1

Loxapine references: 1 2, 13, 14, 15, 16, 17, 18, 19, 20, 21

Clothiapine references: 15, 16, 19, 21 , 22, 23, 24, 25, 26

Miscellaneous references over sulfonates: 27, 28, 29, 30, 31

Sufonate patent application reference: 32

Summary of the Invention

This invention is related to novel iso-Clozapine, Loxapine and Clothiapine analogues of Formula 1 :

or pharmaceutically acceptable acid addition salts thereof, wherein R* is H, (C*-C ₈ ) alkyl or haloalkyl (e. g. CF ₃ (triflates) and CF ₂ CF ₃ ) or hydroxyalkyl, alkenyl, alkynyl.

cyclopropylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyi; R ₂ is H, (C* -C ₈ ) alkyl, alkenyl, alkynyl, cyclopropylalkyl or (C _r C ₈ ) haloalkyl (e. g. CH ₂ CF ₃ CH ₂ CH ₂ CF ₃ , CH ₂ CH ₂ F,

CH ₂ CH ₂ CH ₂ F), hydroxyalkyl, hydroxyalkyloxyalkyl (e. g. or 1-

(alkyl-2-imidazolidinone) (e. g. 1 -(CH ₂ CH ₂ -2-imidazolidinone) and 1 -(CH ₂ CH ₂ CH ₂ -2- imidazolidinonel); X is NH, NR 0, S, SO, S0 ₂ .

The compounds of this invention possess affinity to one or several receptor systems, e. g. DA (D1 -D4), ol , muscarinic (M1 -M4) and 5-HT (5-HT2A, 5-HT2C and 5-HT7). The central nervous system disorders to be treated with the compounds of the present invention include psychoses/schizophrenia, autism, Tourette's syndrome, restless legs, Huntington's chorea, motion sickness, nausea, vomiting and severe anxiety.

In a preferred embodiment, the invention is directed to compounds of Formula 1 wherein R. is haloalkyl R ₂ is H or CH ₃ and X is NH, 0 or S. A more preferred embodiment are compounds of Formula 1 wherein R - is CF ₃ , R ₂ is H or CH ₃ and X is NH, 0 or S. An even more preferred embodiment are compounds of Formula 1 wherein R * is CF ₃ , R ₂ is CH ₃ and X is NH, O or S. The most preferred compound is the compound of Formula I wherein R _* is CF ₃ , R ₂ is CH ₃ and X is NH.

An object of the invention is to provide compounds for therapeutic use, especially compounds having a therapeutic activity in the central nervous system. Another object is to provide compounds having an effect on receptors (DA (D1 -D4), α1 , muscarinic (M1 - M4) and 5-HT (5-HT2A, 5-HT2C and 5-HT7)) in mammals including man.

Processes for preparation of these compounds, their pharmaceutical use and pharmaceutical preparations employing such compounds constitute further aspects of the invention.

Detailed Description of the Invention

In appropriate situations, the proper stereochemistry is represented in the structural schemes.

As used herein the term (C _n -C _m ) is inclusive such that a compound of (C _| -C ₈ ) would include compounds of one to 8 carbons and their isomeric forms. The various carbon moieties are defined as follows: alkyl refers to an aliphatic hydrocarbon radical and

- 4 - includes branched or unbranched forms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neo-pentyl, n-hexyl, isohexγl, n-heptyl, isoheptyl, and n-octyl.

Alkenyl refers to a radical of an aliphatic unsaturated hydrocarbon having a double bond and includes both branched and unbranched forms such as ethenyl, 1 -methyl- 1-ethenyl, 1- propenyl, 2-propenyl, 1 -butenyl, 2-butenyl, 3-butenyl, 2-methyl-1-butenyl, l-pentenyl, 2- pentenyl, 3-pentenyl, 4-pentenyl, 1 -methyl-4-pentenyl, 3-methyl-1-pentenyl, 3-methyl-2- pentenyl, 1 -hexenyl, 2-hexenyl, 3-hexenyl , 4-hexenyl, 1 -methyl-4-hexenyl, 3-methyl-1 - hexenyl, 3-methyl-2-hexenyl, 1 -heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 1 -methyl-4- heptenyl, 3-methyl-1 -heptenyl, 3-methyl-2-heptenyl, 1 -octenyl, 2-octenyl or 3-octenyl. Cγcloalkyl refers to a radical of a saturated cyclic hydrocarbon such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexγl, cycloheptvl, or cyclooctyl. "Halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine.

It will be apparent to those skilled in the art that compounds of this invention may contain chiral centers (e. g. R* and/or R ₂ ). The scope of this invention includes all enantiomeric or diastereomeric forms of Formula 1 compounds, either in pure form or as mixtures of enantiomers or diastereomers. The compounds of Formula 1 can contain one asymmetric carbon atom in the ring system (when X = SO). The therapeutic properties of the compounds may to a greater or lesser degree depend on the stereochemistry of a particular compound. Pure enantiomers, as well as enantiomeric or diastereomeric mixtures, are within the scope of the invention.

Both organic and inorganic acids can be employed to form non-toxic pharmaceutically acceptable acid addition salts of the compounds of this invention. Illustrative acids are sulfuric, nitric, phosphoric, hydrochloric, citric, acetic, lactic, tartaric, pamoic, methanesulfonic, ethanedisulfonic, sulfamic, succinic, cyclohexylsulfamic, fumaric, maieic and benzoic acid. These salts are readily prepared by methods known in the art.

The compounds of this invention may be obtained by one of the following methods described below, as outlined in the appropriate charts.

In clinical practice the compounds of the present invention will normally be administered orally, rectally or by injection, in the form of pharmaceutical preparations comprising the active ingredient either as a free base or as a pharmaceutically acceptable non-toxic, acid addition salt, such as the hydrochloride, lactate, acetate, methanesulfonate, sulfamate

salt, in association with a pharmaceutically acceptable carrier. The use and administration to a patient to be treated in the clinic would be readily apparent to a person of ordinary skill in the art.

s In therapeutical treatment the suitable daily doses of the compounds of the invention are from about 0.1 mg to 2000 mg for oral application, preferably 0.5-500 mg, and 0.05 mg to about 100 mg for parenteral application, preferably 0.05-50 mg daily doses. The daily dose will preferably be administrated in individual dosages 1 -4 times daily and the dosage amounts are based on an individual having a weight of 70 kg. w

The compounds of Formula 1 of this invention, wherein R« is H, (C*-C ₈ ) alkyl or haloalkyl (e. g. CF ₃ (triflates) and CF ₂ CF ₃ ) or hydroxyalkyl, alkenyl, alkynyl, cyclopropylalkyl, aryl, arγlalkγl, heteroaryl, heteroarylalkyl; R ₂ is H, (C ₁ -C ₈ ) alkyl, alkenyl, alkynyl, cyclopropylalkyl or (C _r C ₈ ) haloalkyl (e. g. CH ₂ CF ₃ CH ₂ CH ₂ CF ₃ , CH ₂ CH ₂ F, is CH ₂ CH ₂ CH ₂ F), hydroxyalkyl, hydroxyalkyloxyalkyl (e. g. or 1-

(alkyl-2-imidazolidinone) (e. g. 1 -(CH ₂ CH ₂ -2-imidazolidinone) and 1 -(CH ₂ CH ₂ CH ₂ -2- imidazolidinone)); X is NH, NR., 0, S, SO, S0 ₂ , display binding affinity at the receptor systems: DA (D1-D4), α1 , muscarinic (M1 -M4) and 5-HT (5-HT2A, 5-HT2C and 5-HT7) in mammals including man. These compounds are particularly effective atypical anti- 0 psychotic (anti-schizophrenic) agents. Other uses for these compounds include autism, Tourette's syndrome, restless legs, Huntington's chorea, motion sickness, nausea, vomiting and severe anxiety.

The compounds of this invention also have high oral potency and a long duration of 5 action.

The utility of the compounds of this invention to treat central nervous system disorders can be shown in behavioral and biochemical activity in rats.

0 Effect on DA release in rats as measured bv microdialvsis in freely moving animals.

The biochemical effects of the compounds of this invention can be monitored with the microdialγsis technique in freely moving animals (see under Experimental Section below).

5

Monitoring potential catalepsy in rats,

The propensity of the compounds of this invention to induce catalepsy was measured in rats, who were placed temporarily on a vertical grid (see under Experimental Section below).

Monitoring of potential inhibition of locomotor activity in rats treated with Apomorphine or Amphetamine.

The ability of the componds of this invention to reduce the Apomorphine or Amphetamine induced increase in locomotor activity was measured in rats with motilitγ meters (see under Experimental Section below).

Experimental Section

Syntheses

Starting materials (the corresponding phenols) for the claimed target compounds may be obtained by the methods known in the art.

' H and ¹³ C NMR spectra were recorded at 200 and 50.3 MHz, respectively, on a Varian Gemini 200 spectrometer. CDCI ₃ was employed as the solvent unless otherwise stated. Chemical shifts are given in units (ppm) and relative to TMS or deuterated solvent. IR spectra were obtained on a ATI-Mattson spectrometer. Elemental analyses were performed in the Micro Analytical Department of the University of Groningen or at other approved laboratories. The chemical ionization (CD mass spectra were obtained on a Finnegan 3300 system or a UNICAM Automass 150 GC/MS system, equipped with a UNICAM GC, 610 Series. Melting points were determined on a Electrothermal digital melting point apparatus and are uncorrected. Specific optical rotations were measured in methanol (c = 1.0) at 21 °C on a Perkin Elmer 241 polarimeter.

All methoxylated starting compounds were prepared according to the literature procedures. ^{10, 18, 21 , 33, 34} Chemicals used were commercially available (Aldrich and/or Sigma) and were used without further purification.

Parts of the synthetic methods, not necessarily the preferred route, for forming the sulfone esters from the corresponding phenols are exemplified in Scheme 1 :

Scheme 1

Suitable starting materials, either the phenols or the OMe-analooues. which can be dealkylated to the corresponding phenols with e. g. AICI ₃ in ethylmercaptane at room temperature, for this invention are known:

2-Hydroxy-1 1 -(4-methyl-1 -piperaziπyl)-5H-dibenzolb,e][1 ,4]diazepine; 2-hydroxy-8- dechloroclozapine; RN [156632-07-4]

2-Methoxy- 1 1 -(4-methγl-1 -piperazinyl)-5H-dibenzolb,e][ 1 ,4]diazepine; 2-methoxy-8- dechloroclozapine; RN [95316-97-5]

2-Methoxy-1 1 -(4-methyl-1 -piperazinyl)-dibenzo[b,f][1 ,4]oxazepine; RN [13745-79-4]

2-Methoxy-11-(4-methyl-1-piperazinyl)-dibenzo[b,f][1,4]thiaz epine; RN [13745-79-4]

The following intermediates, useful for the preparation of compounds of the present invention are known:

2-Methoxγ-dibeπz[b,f][1,4]oxazepin-11(10H)-one RN [60287-33-4] ³⁵

2-Hydroxy-dibenz(b,f][1,4]oxazepin-11(10H)-one RN [60287-08-3] ³⁵

2-Methoxγ-dibenzolb,f][1,4]thiazepin-11(10H)-one RN [3158-77-8] ^{36, 37}

Scheme 4

Scheme 5

Synthesis of 2-Trifluoromethanesulfonyloxy-1 1-(4-methyl-1 - piperazinyl)-5H- dibenzo[b,e][1 ,4]diazepine (12).

2-(2-Nitrophenyl)amino-5-methoxybenzoic acid (3). A mixture of 5-methoxγ-2- aminobenzoic acid (1 , 6.68 g, 40 mmol), 2-bromo-nitrobenzene (2, 8.08 g, 40 mmol) and 10 mL of amγl alcohol was heated to 80 °C with stirring. To the homogeneous solution was immediately added copper powder (60 mg) and K ₂ C0 ₃ (5.7 g). The reaction temperature was increased in order to distill off the water generated in the reaction and the amγl alcohol. The reaction mixture was then kept at 210 °C for 3 h. After cooling, the reaction mixture was quenched with 200 mL of water and 100 mL of 4 N aq HCI at room temperature. The precipitate was removed by filtration and the solution was extracted with ethyl acetate. The organic layers were combined and concentrated. The residue was purified by flash chromatography (Si0 ₂ , ethyl acetate as eluent) to afford 6.8 g (59 %) of the title compound.

2-(2-Arninophenyl)amino-5-methoxybenzoic acid (4). Compound 3 [2-(2-nitrophenyl)amino- 5-methoxybenzoic acid] ( 1 .8 g) in THF (50 mL) was hydrogenated with a catalytic amount of palladium on carbon in a Parr-shaker apparatus under 4.5 atm H ₂ (g) at 60 °C for 4 h to afford 1 .4 g (88 %) of the title compound.

2-Methoxy-5,10-dihydro-1 1 -oxo-dibenzo(b,e][1 ,4]diazepine (5). Intermediate 4 [2-(2- aminophenyl)amino-5-methoxybenzoic acid] (3.0 g) in 100 mL of xylene was refluxed for 40 h. After evaporation the residue was purified by flash chromatography (Si0 ₂ , hexane and ethyl acetate, 4: 1 then 1 : 1 , as eluents) to afford 2.2 g (79 %) of the title compound. MS (El) shows M ⁺ at m/e = 240.

2-Methoxy-1 1 -(4-methyl-1 -piperazinyl)-5H-dibenzo[b,el[1.4l-diazepine (7, RN [95316-97- 51). Intermediate 5 (200 mg, 0.83 mmol), phosphorus oxychloride (POCI ₃ ) (3 mL), toluene (5 mL) and N,N-dimβthylaniline (0.5 mL) were combined and heated to reflux for 3 h. The solvents of the mixture were evaporated under vacuum to afford the iminochloride intermediate 6, which was used in the next-step reaction without further purification.

Above iminochloride 6 in 3 mL of toluene and 2 mL of N-methylpiperazine were refluxed for 3 h. After evaporation the residue was taken up with chloroform, washed with 2N aq NaOH and purified by flash chromatography (Si0 ₂ , ethyl acetate then THF as eluents) to afford 214 mg (80 %) of the title compound. MS (El) shows M ⁺ at m/e = 322.

2-Hydroxy-11-(4-methyl-1-piperaziπyl)-5H-dibenzo[b,e][1,4]- diazepine (8, RN [156632-07- 4]). 2-Methoxy-1 1-(4-methγl-1-piperazinyl)-5H-dibeπzo[b,el[1 ,4]diazepine (7) (200 mg, 0.62 mmol) in 3 mL of EtSH was treated with AICI ₃ (1 .5 g) with stirring at room temperature for 4 h. The mixture was quenched with 30 mL of ice-water. The pH value of 5 the solution was adjusted to pH 8 with 2N aq NaOH. The solution was extracted with chloroform (4 x 30 mL). The organic layers were combined, dried over MgS0 ₄ filtered, and the solvents were evaporated. The light yellow solid residue was recrystallized from ethyl acetate to afford 140 mg (73 %) of the title compound as fine crystals: mp 266 °C, with all of the expected spectra including IR, -H NMR, ^{1 3} C NMR. MS (El) shows M at w m/e = 308.

2-Hydroxy-5,10-dihydro-1 1 -oxo-dibenzo[b,e][1 ,4]diazepine (9). 2-Methoxy-5,10-dihγdro- 1 1 -oxo-dibenzo[b,eH1 ,4]diazepine (5) (600 mg, 2.5 mmol) in 4 mL of EtSH was treated with AICI ₃ (1 .6 g) with stirring at room temperature for 4 h. The mixture was quenched 15 with 20 mL of ice-water and then 10 mL of 4N aq HCI. The solution was extracted with chloroform (2 x 20 mL) and ethγlacetate (2 x 20 mL). The combined organic layers were dried over MgS0 ₄ , filtered, and the solvents were evaporated. The solid residue was recrystallized from ethyl acetate to afford 505 mg (88 %) of the title compound as fine crystals: mp 280 °C, with all the expected spectra including IR, ^* H NMR, ^{1 3} C NMR. MS 0 (El) shows M at m/e = 226.

2-Trifluoromethanesulfonyloxy-5,10-dihydro-11-oxo-dibenzo -[b,e][1 ,4]diazepine (10). 2- Hydroxy-5, 10-dihydro-1 1-oxo-dibenzo[b,e][1 ,4]-diazepine (9) (400 mg, 1.77 mmol) in 10 mL of dioxane was treated with N-phenyl-trifluoromethanesulfonimide (800 mg) in the 5 presence of 2 mL of triethylamine with strirring at room temperature overnight. After evaporation of the solvents, the residue was purified by flash chromatography (Si02, hexane and ethyl acetate, 9:1 then 4: 1 , as eluents). Recrystallization from hexane and ethyl acetate (10: 1 ) afforded 380 mg (60 %) of the title compound as light yellow crystals. MS (El) shows M ⁺ at m/e = 358. 0

2-Trifluoromethanesulfonyloxy-11-(4-methyl-1-piperazinyl) -5H-dibenzo[b,e][1 ,4]diazepine (12). Method A. 2-Hydroxy-1 1 -(4-methyl-1 -piperazinyl)-5H-dibenzo[b,e][1 ,4]diazepine (8) (180 mg, 0.58 mmol) in 10 mL of CH ₂ CI ₂ was treated with N-phenyl- trifluoromethanesulfonimide (300 mg, 0.83 mmol) in the presence of 1 mL of 5 triethylamine with strirring at room temperature overnight. After evaporation the residue was purified by flash chromatography (Si0 ₂ ; hexane and ethyl acetate, 1 :1 ; then ethyl

acetate as eluents). Recrγstallization from hexane and ethyl acetate (15:1 ) afforded 200 mg (78 %) of the title compound as fine crystals: mp 160 °C, with the expected spectra data including IR, ¹ H NMR, ^{1 3} C NMR. MS (El) shows M ⁺ at m/e = 440.

Method B. 2-Trifluoromethanesulfonyloxy-5,10-dihydro-1 1-oxo-dibenzo-[b,e][1 ,4]diazepine (10) (260 mg, 0.72 mmol), phosphorus oxychloride (P0CI ₃ ) (4 mL), toluene (10 mL) and N,N-dimethylaniline (0.5 mL) were combined and heated to reflux for 3 h. The mixture was evaporated under vacuum to afford the corresponding iminochloride intermediate 11 , which was used in the next-step reaction without further purification.

The iminochloride 11 in 10 mL of toluene was treated with 3 mL of N-methylpiperazine under refluxing for 3 h. After evaporation of the solvents, the residue was taken up in chloroform, washed with 2N aq NaOH and purified by flash chromatography and recrystallized, as described in Method A above, to afford 230 mg (73 %) of the title compound 12.

2-Methanesulf onyloxy-1 -(4-methyl-1 -piperazinyl)-5H-dibenzo-[b.e][ 1 ,4]diazepine (15).

2-Methanesulfonyl-5,10-dihydro-11-oxo-dibenzo[b,e][1 ,4]diazepine (13). 2-Hydroxy-5,10- dihydro-1 1 -oxo-dibenzo[b,e][1 ,4]diazepine (9) (50 mg, 0.22 mmol) in 6 mL of dichloromethanβ was treated with methanesulfonylchloride (34 mg, 0.30 mmol) in the presence of 0.2 mL of triethylamine at -78 °C. The mixture was stirred for 1 h, in order to allow the reaction temperature to reach room temperature, quenched with water and extracted with chloroform. Workup afforded 50 mg (76 %) of the title compound. MS (El) shows M ⁺ at m/e = 304.

2-Methanesulfonyloxy-11-(4-methyl-1-piperazinyl)-5H-dibβ nzo[b.e][1,4]diazepine (15). 2- Methanesulfonyloxy-5,10-dihydro-1 1-oxo-dibenzo(b,el[1 ,4]diazepine (13) (50 mg, 0.16 mmol), phosphorus oxychloride (POCI ₃ ) (2 mL), toluene (3 mL) and N,N-dimethylaniline (0.1 mL) were combined and heated to reflux for 3 h. The mixture was evaporated under vacuum to afford the iminochloride intermediate 14, which was used in the next reaction step without further purification.

This iminochloride 14 in 3 mL of toluene was treated with 1 mL of N-methylpiperazine under refluxing for 3 h. Similar workup as described for the preparation of 12 afforded 32 mg (50 %) of the title compound as fine crystals: mp 173-177 °C. MS (El) shows M ⁺ at m/e = 386.

Preparations of 2-(4-methylphenylsulfonyloxy)-1 1-(4-methγl-1-piperazinyl|-5H- dibenzo[b,e][1 ,4]diazepine (16), 2-phenγlsulfonyloxy-1 1-<4-methyl-1 -piperazinyl)-5H- dibenzo[b,e][1 ,4]diazepine (17) and 2-(2-thienylsulfonyloxy)-1 1 -(4-methγl-1 -piperazinyl)- 5H-dibenzo[b,e][1 ,4)diazepine (18) were performed by the same procedure as for the preparation of 2-trifluoromethanesulfonyloxγ-1 1 -(4-methyl-1 -piperazinyl)-5H- dibenzo[b,e][1 ,4]diazepine (12) (Method A) from 2-hydroxy-1 1 -(4-methyl-1 -piperazinyl)- 5H-dibenzo[b,e][1 ,4]diazepine (8), however, using the sulfonation procedure with appropriate sulfonylchloride, as described for the preparation of compound 13.

Mass spectrometrγ:

Compound no M ⁺ at m/e

16 462

17 448 18 454

2-Tπfluoromethanesulf onyloxy-11 -(4-methylpiperazino)-5H-dibβnzo[b,f ][ 1 ,4)oxazepine ( 19)

(Scheme 4). 2-Methoxy-5,10-dihydro-1 1 -oxo-dibenzo[b,f][1 ,4]oxazepine (RN [60287-33- 4]) was prepared according to the literature. 35

2-Hydroxy-5,10-dihydro-1 1 -oxo-dibenzo[b,f][1,4loxazepine (RN [60287-08-3] ). 2- Methoxy-5,10-dihγdro-1 1 -oxo-dibenzo[b,e][1 ,4]oxazepine (1.2 g) in 10 mL of EtSH and 10 mL of dichloromethane was treated with AICI ₃ (5.0 g) with stirring at room temperature for 6 h. The solvent of the mixture was evaporated by rotavapor at reduced pressure, quenched with 20 mL of ice-water and then 20 mL of 4N aq HCI. A white precipitate was collected, washed with water and dried under vacuum. The product (1 .0 gram) showed the expected spectra including IR, 'H NMR, and MS (El, m/z 227).

2-Trifluoromethanesulfonγloxy-5,10-dihydro-11 -oxo-dibenzo[b,f][1 ,4]-oxazepine. 2- Hydroxy-5,10-dihydro-1 1 -oxo-dibenzo[b,e][1 ,4]oxazepine (1 .0 g) in 20 mL of dichloromethane and 5 mL of triethylamine was cooled to -78 °C, treated with triflic anhydride (1.54 mL, 7 mmol). The mixture was stirred for 3 h to allow the temperature back to room temperature, quenched with 3 mL of 50% methanol in water and concentrated. The residue was flashed on silical-gel column (hexane and ethyl acetate, 9:1 then 1 :1 , as eluents) to afford 1.3 g of the title compound as white crystal: MS (El) m/z 359.

2-Trifluoromethanesulfonyioxy-1 1 -(4-methylpiperazino)-5H-dibenzo-[b,f][1 ,4]oxazepinβ. 2- Trifluoromethanesulfonyl-5,10-dihydro-1 1-oxo-dibenzo-[b,e][1 ,4]oxazepine (1 .0 g), P0CI ₃ (5 mL), toluene (10 mL) and N,N-dimethyl-aniline (1.0 mL) were combined and heated to reflux for 3 h. The solvent of the mixture was evaporated under vacuum to afford iminochloride intermediate, which was dissolved in 10 mL of toluene and was treated with 5 mL of N-methylpiperazine under refluxing conditions for 3 h. After evaporation the residue was purified by flash chromatography (Si0 ₂ , 9:1 hexane and ethyl acetate then pure ethylacetate as eluents) and recrystallization (n-hexane) to afford 1 .01 g of the title compound: mp 1 17-1 18 °C; MS(EI) m/z 441 ; ¹ H NMR (CDCI ₃ ) and elements (C. H. N) were analyzed.

2-Trifluoromethanesulfonyloxγ-11-(4-methylpiperazino)-5H -dibenzo-[b,f][1,4]thiazepine (20) (Scheme 5). 2-[(4-Methoxyphenyl)thio]benzoic acid (RN 19862-91 -0) was prepared according to the literature.

2-[(4-methoxyphenyl|thio]benzoyl azide. 2-[(4-methoxγphenyl)thiolbenzoic acid (14 g, 53.8 mmol) was treated with 50 mL of S0CI ₂ and 1 mL of DMF at refuxing for 1 h. The mixture was evaporated under vacuum and the solid residue was dissolved into dry acetone (100 mL). The resulting solution was added dropwise to a cooled 30 % aq NaN ₃ (60 mL) during 30 min. The suspension was stirred at 0 °C for 1 h, diluted with water

(400 mL), filtered, washed well with water and dried overnight under vacuum to afford 14 g of the title compound as a fine solid. IR and GC-MS were analyzed. Mass spectra (El) observed m/z at 257 (M ⁺ -N ₂ , isocyanation product under GC conditions: 150-300 °C/10

°C rate program) conditions.

2-Hydroxy-dibenzo[b,f][1,4]thiazβpinβ-11(10H)-one. 2-[(4-methoxγ-phenyl)thio)benzoyl azide (1 .4 g) in 20 mL of dichlorobenzene was added to a suspension of AICI ₃ (2.7 g) in o-dichlorobenzene (50 mL) with stirring. The mixture was heated to reflux for 15 min, cooled to room temperature. CHCI ₃ (150 mL) was added, extracted with 4 N aq HCI (2 x 50 mL). The organic layer was concentrated under vacuum and the residue was purified by flash chromatography (Si0 ₂ , ethyl acetate as eluent) to afford 240 mg of the title compound. Mass spectra observed m/z at 243 (M ⁺ ).

2-Triflatβ-dibenzo[b,f][1,4]thiazepine-11(10H)-one. 2-Hydroxγ-dibenzo-[b,f][1 ,4]thiazepine- 1 1 (10H)-one (100 mg) in CH ₂ CI ₂ was cooled to -78 °C and treated with triflic anhydride (0.13 mL) in the presence of triethylamine (1 .0 mL). After stirring for 2 h the mixture was

mixed with Si0 ₂ (3 mL), evaporated and flashed (Si0 ₂ column, hexane and ethyl acetate as eluents) to afford 130 mg of the title compound. MS (El) at m/z 375 (M ⁺ ).

2-Trif luoromethanesulf onyloxy- 1 1 -(4-methhylρiperazino)-5H-dibenzo-[b ,f ] [ 1 ,4]thiazepine . 2-Trifluoromethanesulfonyloxy-5, 10-dihydro-1 1 -oxo-dibenzo-[b,e][1 ,4)thiazepine (80 mg), POCI ₃ (1 mL), toluene (1 mL) and N,N-dimethyl-aniline (0.1 mL) were combined and heated to reflux for 3 h. The solvent of the mixture was evaporated under vacuum to afford iminochloride intermediate, which was dissolved in 1 mL of toluene and was treated with 1 mL of N-methylpiperazine under refluxing conditions for 3 h. After evaporation the residue was purified by flash chromatography (Si0 ₂ , 9:1 hexane and ethyl acetate then pure ethγlacetate as eluents) and recrγstallization from hexane to afford 33 mg of the title compound: MS(EI) m/z 457; ¹ H NMR (CDCI ₃ ) and IR were analyzed.

Derivatives with the -CH ₂ CH ₂ -2-imidazolidinone and -CH ₂ CH ₂ CH ₂ -2-imidazolidinone substutυent R ₂ (see Formula I).

These derivatives are synthesized from the corresponding N-methyl-piperazine derivatives (e. g. compounds 12, 15-20) by first performing a demethγlation with 1 -chloroethyl chloroformate ³⁹ and then an alkγlation with the appropriate alkγlating agent (e. g. 1 -(2- chloroethγl)-2-imidazoiidinone and 1 -(3-chloroprophyl)-2-imidazolidinone, respectively), refluxing over night in methyl isobutyl ketone (MIBK) in the presence of dry, ground K ₂ C0 ₃ (s) and a catalytic amount of Kl (s). ¹

Pharmacology

Microdialysis experiments: Adult male Wistar rats (Harlan Centraal Proefdier Bedrijf, The Netherlands) were used. Until implantation of the dialysis probe, the rats were housed in groups of 5-10 animals in plastic cages. The rats were kept in a room maintained under constant temperature (21 °C) and humidity (40%) on a 12:12 dark:light cycle (07:00 on and 19:00 off) with food and water available ad libitum. The rats weighed between 250- 350 g at the time of the microdialysis probe implantation. Animal procedures were conducted in accordance with guidelines published in the NIH Guide for Care and Use of Laboratory Animals and all protocols were approved by an Institutional Animal Care and Use Committee.

Drugs were administered s.c. in a volume of 1 mL/kg in distilled water. Sometimes a drop of acetic acid was added to improve the solubility of the test compound.

The microdialysis probes used in the present investigation were of a vertical, concentric design: A piece of fused silica capillary (Polymicro Technologies, Phoenix, AZ, USA) with I.D. 50 μm and O.D. 1 15 μm was threaded through a piece of PE-20 tubing (Clay Adams, Parsippanγ, NJ, USA) 0.5 cm from the end. A dialysis membrane (8 mm) (Hospal, Meyzieu, France) with an I.D. of 250 μm and an O.D. of 300 μπrt was slipped over the fused silica, glued into the PE-20, and closed at the end with epoxy (Devcon, Danvers, MA, USA). The fused silica was then brought into close apposition to the closed end of the dialysis membrane and fixed into position by epoxy at the junction of the silica and PE- 20. The active membrane region was 2 mm in length and the region above the active area was coated with a thin layer of epoxy. After cutting the silica and PE-20 tubing 5 mm from the junction of the silica and PE-20, an inlet (a 25 G stainless steel needle that was broken at the bevel and from the plastic fixture) was glued with epoxy over the silica. The other end of the PE 20 served as the outlet.

The animals were anesthetized with chloral hydrate (400 mg/kg) and a microdialysis probe was implanted into the CPu at the following coordinates: (flat skull) AP 1.0 mm, ML + 2.5 mm relative to bregma, and -6.0 mm below the dura and/or nucleus accumbens at the following coordinates: (flat skull) A. P. 2.5 mm, ML -1.3 relative to bregma, and -7.3 mm below the dura. The probes were lowered using an electrode carrier (Model 1760, David Kopf, Tujunga, CA, USA) at a speed of approximately 400 μm/miπ and secured to the skull with fast-curing dental cement on three skull screws. At the start of the experiment, 16 hours after surgery, artificial cerebrospinal fluid (aCSF) consisting of (mmol/L): NaCI 147, KCI 2.5, CaCI ₂ 1 .3, and MgCI 0.9, pH 7.4 was perfused through the probe with a syringe pump (Harvard Apparatus inc, South Natick, MA, USA). The outlet of the probes were connected to the valve of an HPLC apparatus and samples were collected on-line and injected every 15 min from a 20 μ sample-loop. Dopamine, DOPAC, and 5-HIAA were quantified in the samples according to previously described methods. Before initiating any pharmacological manipulation, baseline samples were taken until there was less than 30% variation between the samples.

Behavioral experiments: Adult male Wistar rats (Harlan Centraal Proefdier Bedrijf, The Netherlands) were used. The rats were housed in groups of 12 animals in plastic cages. The rats were kept in a room maintained under constant temperature (21 °C) and humidity (40%) on a 12:12 dark:light cycle (07:00 on and 19:00 off) with food and water available ad libitum. The rats weighed between 200-250 g at the start of the experiment. Animal procedures were conducted in accordance with guidelines published in the NIH Guide for Care and Use of Laboratory Animals and all protocols were approved by an

Institutional Animal Care and Use Committee.

Catalepsy test: Rats were placed in a plastic bowl which measured 30 cm in diameter and 20 cm in height. After 10 minutes, the rats were placed on a vertical metal grid (grids 1 cm square) which measured 15 x 40 cm (width x heigth). The time until the first paw movement and the time to get off the grid were measured in three consecutive sessions. Thereafter, the rat was injected with either saline of test compound and left in the cage for another 30 minutes before catalepsy was retested.

Locomotor activity: Directly after the second catalepsy test (see above), the animals were transferred to a locomotion chamber (Columbus Instruments, Ohio, USA) and locomotion was measured for 10 minutes (referred to as exploratory behavior due to the new environment). Apomorphine (1 mg/kg, s.c.) was subsequently injected and locomotion was measured for 30 minutes after the injection.

Results

TABLE 1. Summary of the pharmacological characterization of compound 12, 15, 19, 20, clozapine, olanzapine, and haloperidol.

Footnotes Table 1 : Interaction with MK801 : Rats were injected with test compounds 15 min prior to the administration of 0.25 mg/kg MK801 in their home cages. Immediately after the administration of MK801 , the rats were transferred to a locomotor chamber (Columbus Instruments, Ohio, USA) and locomotion was measured for 30 min.

Interaction with AMPHETAMINE: Rats were injected with thest compounds 15 min prior to the administration of 0.5 or 2 mg/kg amphetamine in their home cages. Immediately after the administration of amphetamine, the rats were transferred to a locomotor chamber (Columbus Instruments, Ohio, USA) and locomotion was measured for 60 min.

References

1 . Liegeois, J. F. F.; Rogister, F. A.; Bruhwyler, J.; Damas, J .; Nguyen, T. P.; Inarejos, M. 0.; Chleide, E. M. G.; Mercier, M. G. A.; Delarge, J. E.

Pyridobenzoxazepine and Pyridobenzothiazepine Derivatives as Potential Central Nervous System Agents: Synthesis and Neurochemical Study. J. Med. Chem. 1994, 37, 519-25.

2. Carlsson, A.; Lindqvist, M. Effect of Chlorpromazine or Haloperidol on formation of 3-methoxytyramine and norepinephrine in the mouse brain. Acta Pharmacol.

Scand. 1963, 20, 140-144.

3. Snyder, S. H. DA hypothesis of schizophrenia. Am. J. Psychiatry 1976, 133, 197.

4. Sedvall, G. Monoamines and schizophrenia. Acta Psychiat. Scand. 1990, 82 (Sυppl. 358), 7-13.

5. Povlsen, U. J.; Noring, U.; Fog, R.; Gerlach, J. Tolerability and Therapeutic effect of Clozapine. Acta Psychiatr. Scand. 1985, 71, 176-185.

6. Phillips, S. T.; De Paulis, T.; Baron, B. M.; Siegel, B. W.; Seeman, P.; Van Tol, H. H. M.; Guan, H.-C; Smith, H. E. Binding of 5H-Dibenzo[b,e][1 ,4)diazepine and Chiral 5H-Dibenzo[a,d)cγcloheptene Analogues of Clozapine to Dopamine and

Serotonin Receptors. J. Med. Chem. 1994, 37, 2686-2696.

7. Marder, S. R.; Van Putten, T. Who Should Receive Clozapine? Arch. Gen. Psychiatry 1988, 45, 865-867.

8. Bϋrki, H. R.; Sayers, A. C; Ruch, W.; Asper, H. Effects of Clozapine and other Dibenzo-Epines on Central Dopaminergic and Cholinergic Systems. Structure- Activity Relationships. Arzneim.-Forsch. 1977, 27, 1561 -1565.

9. Howell, C. F.; Hardy, R. A. J.; Quinones, N. Q. Patent: 1 1 -Tertiary-

- 24 - aminodibenz[b,f][ 1 ,4]oxazepines and thiazepines <FR 1 508536 680105 PRAI US 6601 17 (20 pp.)). Fr. Patent 1966.

10. Schmutz, J .; Hunziker, F.; Kuenzle, F. M. Patent: Substituted dibenz[b,f][1 ,4]oxazepines (CH 436297 671 1 15 (3 pp.)). Swiss Patent Coden: Swxxas 1964.

1 1 . Schmutz, J.; Hunziker, F.; Fischer, E. Patent: Diazepine and thiazepine derivatives. S. African patent application, 29 pp., PI ZA 7106668 730405, Al ZA 71-6668 71 1005 1971.

12. Krusteva, I.; Vitev, M.; Denkova, E.; Georgiev, A.; Evstatieva, A.; Zikolova, T. Synthesis of 2-chloro- 1 1 -(4-methyl- 1 -piperazinγl)dibenz[b,f ][ 1 ,4]oxazepine succinate. I. Preparation of 2-chloro-1 1 -(4-methyl-1 - piperazinyl)dibenz[b,fl[ 1 ,4]oxazepine. Farmatsiya (Sofia) 1987, 37, 1 -6.

13. Izquierdo, S. M.; Martin, E. P. U. Patent PI ES 490495 A1 810516, Al ES 80- 490495 80041 1 : Dibenzo[b,f][ 1 ,4]oxazepine and its salts. Span. Patent 1980. 14. Howell, C. F.; Hardy, R. A. J.; Quinones, N. Q. Patent PI JP 51041639 B4

761 1 1 1 Showa, Al JP 67-3069 6701 17: 1 1 -Aminodibenz[b,f][ 1 ,4]oxazepines and -thiazepines. Japan. Patent 1967.

1 5. Schneider, J. Patent PI DE 2316438 73101 1 , PRAI CH 72-4898 720404: Piperazinγl-substituted dibenzoxazepines and similar compounds. Ger. Often. Patent 1972.

16. Sheely, R. M.; Allen, G. R. J. Patent PI GB 1 164360 690917, PRAI US 661 21 6: Analgesic, antidepressant and tranquillizing 1 1 - aminodibenz[b,f][1 ,4]oxazepines and -thiazepines. Brit. Patent 1966.

1 7. Howell, C. F.; Hardy, R. A. J.; Quinones, N. Q. Patent PI US 3444169 690513, Al US 6601 17: Tranquilizing and antidepressant 1 1 - aminodibenz[b,f][1 ,4]oxazepines and thiazepines. U.S. Patent 1966.

18. Howell, C. F.; Hardy, R. A. J.; Quinones, N. Q. Patent PI FR 1508536 680105, PRAI US 6601 17: 1 1 -Tertiary-aminodibeπz(b,f][ 1 ,4]oxazepines and thiazepines. Fr. Patent 1966, 1 9. Coppola, J. A. Patent PI US 3412193 681 1 19, Al US 651213: 1 1 -(4-Methyl-

1 -piperazinyl)dibeπz[b,f][1 ,4]oxazepines or -thiazepines. U.S. Patent 1965.

20. Schmutz, J.; Hunziker, F.; Kuenzle, F. M. Patent PI CH 436297 671 1 15, Al CH 640527: Substituted dibenz[b,f][1 ,4]oxazepines. Swiss Patent 1964.

21 . Schmutz, J.; Hunziker, F.; Stillβ, G.; Lauener, H. (Clothiapine-OMe ref) Chemical structure and pharmacological activity of a new group of tricyclic neuroleptics. XI.

Heterocycles with seven atoms. Chim. Ther 1967, 2, 424-9.

22. Nagarajaπ, K.; Kulkarni, C. L.; Venkateswarlu, A.; Shah, R. K. Condensed heterotricycles. Dibenz[b,f][1 ,4]oxazepin-1 K10H)- thiones, 1 1 -substituted dibenz[b,f][1 ,4]oxazepines, and dibenz[b,f][1 ,4]thiazepine analogs. Indian J. Chem 1974, 12, 258-62. 23. Schmutz, J.; Hunziker, F.; Fischer, E. Patent PI ZA 7106668 730405, Al ZA

71 -6668 71 1005: Diazepine and thiazepine derivatives. S. African Patent 1971. 24. Schmutz, J.; Hunziker, F. Patent PI CH 476753 690930, Al CH 620608:

Analgesic and sedative 1 1 -(substituted-amino)dibenzo[b,f][1 ,4]thiaze pines. Swiss Patent 1962. 25. Howell, C. F.; Hardy, R. A. J.; Quinones, N. Q. Patent: PI FR 1508536 680105, PRAI US 6601 17: 1 1 -Tertiarγ-aminodibenz[b,f][1 ,4]oxazepines and thiazepines. Fr. Patent 1966. 26. Schmutz, J.; Hunziker, F.; Fischer, E. Patent PI CH 422793 670429, Al CH 610720: Dibenzo[b,f][1 ,4]thiazepines. Swiss Patent 1961. 27. Sonesson, C; Lin, C. H.; Hansson, L.; Waters, N.; Svensson, K.; Carlsson, A.;

Smith, M. W.; Wikstrom, H. Substituted (S)-Phenγlpiperidines and Rigid Congeners as Preferential Dopamine Autoreceptor Antagonists: Synthesis and Structure- Activity Relationships. J. Med. Chem. 1994, 37, 2735-53.

28. Haadsma-Svensson, S. R.; Smith, M. W.; Lin, C.-H.; Duncan, J. N.; Sonesson, C; Wikstrom, H.; Waters, N.; Carlsson, A.; Svensson, K. Synthesis and biological activity of C-5 modified derivatives of ( + )-AJ76 and ( + )-UH232: increased dopamine D3 receptor preference and improved pharmacokinetic properties. Bioorg. Med. Chem. Lett 1994, 4, 689-94.

29. Sonesson, C; Boije, M.; Svensson, K.; Ekman, A.; Carlsson, A.; Romero, A. G.; Martin, I. J.; Duncan, J. N.; King, L. J.; Wikstrom, H. Orally active central dopamine and serotonin receptor ligands: 5-, 6-, 7-, and 8- [[(trifluoromethyl)sulfonyl]oxy]-2-(di-n- propylaminoltetralins and the formation of active metabolites in vivo. J. Med. Chem. 1993, 36, 3409-16.

30. Sonesson, C; Waters, N.; Svensson, K.; Carlsson, A.; Smith, M. W.; Piercey, M. F.; Meier, E.; Wikstrom, H. Substituted 3-phenylpiperidines: new centrally acting dopamine autoreceptor antagonists. J. Med. Chem. 1993, 36, 3188-96.

31. Svensson, K. A. I.; Wikstrom, H. V.; Carlsson, P. A. E.; Boije, A. M. P.; Waters, R. N.; Sonesson, C. A.; Stjernlόf, N. P.; Andersson, B. R.; Hansson, L. O. Patent PRAI US 91 -686799 910417: Centrally acting substituted phenylazacycloalkanes. Pet Int. Appl 1991.

32. Wikstrom, H. Patent PRAI SE 91 -3745 91 1218: Preparation of (hetero)aryl triflates as nervous system agents. Pet Int. Appl 1991.

33. Schmutz, J.; Kuenzle, G.; Hunziker, F.; Gauch, R. Heterocycles with 7-membered rings. IX. 1 1 - Amino substituted dibenzo[b,f]-1 ,4-thiazepines and -oxazepines. Helv. Chim. Acta 1967, 50, 245-54.

34. Schmutz, J.; Hunziker, F.; Stille, G.; Lauener, H. Chemical structure and pharmacological activity of a new group of tricyclic neuroleptics. XI.

Heterocycles with seven atoms. Chim. Ther. 1967, 2, 424-9.

35. Brewster, K.; Clarke, R. J.; Harrison, J. M.; Inch, T. D.; Utley, D. Preparation of the eight monohydroxydibenz[b,f][ 1 ,4]oxazepin-1 1 (10H)-ones. J. Chem. Soc, Perkin Trans 1976, 7, 1286-90. 36. Bennett, 0. F.; Johnson, J.; Tramondozzi, J. Synthesis of 2- methoxydibenzo[b,f]( 1 ,4)-thiazepin-1 1 (10H)-one 5,5-dioxide. Org. Prep. Proced. Int 1974, 6, 287-93. 37. Schmutz, J.; Hunziker, F.; Schindler, 0.; Kuenzle, F. M. Patent PI US 3367930 680206, Al US 640924: Preparation of dibenzo[b,f][ 1 ,4]thiazepines and dibenz[b,f][ 1 ,4)oxazepines. U.S. Patent 1964.

38. Pelz, K.; Jirkovsky, I.; Adlerova, E.; Metysova, J.; Protiva, M. Collect. Czech.

Chem. Commun. 1968, 33(6), 1895-1910. 39. Olofson, R. A.; Martz, J. T.; Senet, J.-P.; Piteau, M.; Malfroot, T. A New Reagent for the Selective, High-Yield N-Dealkylation of Tertiary Amines: Improved Syntheses of naltrexone and Nalbuphine. J. Org. Chem. 1964, 2081 -2082.