NEW CENTRALLY ACTING 5-, 6-, 7-, AND 8-SUBSTITUTED SULPHONE ESTERS OF N-MONOSUBSTITUTED 2- AMINOTETRALINS AND RELATED STRUCTURES

Field of the Invention

The present invention is directed toward new 5-, 6-, 7-, and 8- sulphone ester N-monosubstituted 2-aminotetralins 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 dopamine (DA) and/or 5-HT1 A and/or 5-HT1 D receptors in mammals.

Background of the Invention.

In depressed patients evidence indicates that the neurotransmission in the central nervous system (CNS) may be disturbed. These disturbances involve the neurotransmitters noradrenaline (NA) and 5-hydroxytryptamine (5-HT). The drugs most frequently used in the treatment of depression are considered to act by improving the neurotransmission of either or both of these physiological agents. The mechanism of action for conventional drugs used to treat mental depression is generally believed to be indirect. It is thought the drugs block the reuptake of the neurotransmitters released from nerve terminals in the CNS,

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NA and/or 5-HT, which increases the concentration of these transmitters in the synaptic cleft and restores an adequate neurotransmission. For example, the clinically documented antidepressive drug, zimelidine (dimethy-amino-1 -(4-bromo- phenyl)-1 -(3-pyridyl)propene) acts as such a reuptake inhibitor with high selectivity for 5-HT neurons.

Available data suggests that the enhancement of 5-HT neurotransmission will primarily improve depressed mood and anxiety, whereas the enhancement of NA neurotransmission will improve retardation symptoms occurring in depressed patients. In recent years many efforts have been made to develop new drugs with high selectivity for the improvement of the 5-HT neurotransmission in the CNS.

A fundamentally different way to improve the neurotransmission in the central 5-HT neurons would be to use a 5-HT receptor agonist acting directly upon the 5-HT receptor, and particularly the 5-HT1 A receptor or, alternatively, to use a 5-HT1 D receptor antagonist. In order to minimize side effects, a high selectivity for either of these receptor subtypes would be necessary.

Clinically, 5-HT1 A agonists have also demonstrated anxiolytic properties. The drug, Buspirone hydrochloride is the only currently available marketed 5-HT1 A agonist displaying anxiolytic activity.

This compound antagonizes DA receptors at the same dose it stimulates 5-HT1 A receptors. A similar drug, Gepirone hydrochloride also has DA antagonist properties. These DA

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antagonist properties reduce the clinical utility of these compounds, because long term treatment with DA antagonists can produce tardive dyskinesias.

The search for new CNS active compounds is focused on finding compounds with selective 5-HT1 A receptor agonist effects without detrimentally influencing central DA receptors.

Hellstrand et al. have shown that serotonin and 5-HT1 A receptor agonists stimulate "killer cell" activity in vitro and in research animals with induced tumors. (Hellstrand, K.; Hermodsson, S. J. Immunol. 1987, 139, 869; Hellstrand, K.; Hermodsson, S. Scand. J. Immunol. 1 990, 32, 1 83; Hellstrand, K.; Hermodsson, S. Cell. Immunol. 1990, 127, 199.)

In recent years a large body of pharmacological, biochemical and electrophysiological evidence has provided considerable support in favor of the existence of a specific population of central autoregulatory DA receptors located in the dopaminergic neuron itself and belonging to the D2 and/or the D3 receptor subclasses of DA receptors. These receptors are part of a homeostatic mechanism that modulates nerve impulse flow and transmitter synthesis and regulates the amount of DA released from the nerve endings.

Drugs acting on central DA transmission are clinically effective in treating a variety of central nervous system disorders such as parkinsonism, schizophrenia, and mano-depressive illness. In

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parkinsonism, for example, the nigro-neostriatal hypofunction can be restored by an increase in postsynaptic DA receptor stimulation. In schizophrenia, the condition can be normalized by achieving a decrease in postsynaptic DA receptor stimulation. Classical antipsychotic agents directly block the postsynaptic DA receptor. The same effect can be achieved by inhibition of intraneuronal presynaptic events essential for the maintenance of adequate neurotransmission, transport mechanism and transmitter synthesis.

Direct DA receptor agonists, like apomorphine, are able to activate the DA autoreceptors (presynaptic) as well as the postsynaptic DA receptors. The effects of autoreceptor stimulation appear to predominate when apomorphine is administered at low doses, whereas at higher doses the attenuation of DA transmission is outweighed by the enhancement of postsynaptic receptor stimulation. The antipsychotic and antidyskinetic effects in man of low doses of apomorphine are likely due to the auto- and/or D3- receptor stimulator properties of this DA receptor agonist. This body of knowledge indicates DA receptor stimulants with a high selectivity for central nervous DA autoreceptors would be valuable in treating psychiatric disorders and extrapyramidal side-effects in patients treated with nuroleptic agents (e.g. with Haloperidol).

Information Disclosure Statement.

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

Arvidsson, L-E, et al., J. Med. Chem., 24, 921 (1 981 ), describes hydroxy-2-aminotetralins where the amine is substituted with one n-propyl, one benzyl or two n-propyl substitutents. The 5-, 6-, and 7-hydroxy compounds are described as active central DA-receptor agonists and the 8-hydroxy compound is described as a central 5- HT receptor agonist devoid of DA receptor stimulating activity.

Arvidsson, L-E, et al., J. Med. Chem., 27, 45 (1984), describes 2- aminotetralins where the amine is substituted with one or two methyl, ethyl, n-propyl, i-propyl, n-butyl, or benzyl substituents. The 2-piperidinyltetralin is also described. Several of these compounds were found to be potent 5-HT agonists devoid of DA- mimetic effects.

Arvidsson, L-E, et al., J. Med. Chem., 30, 2105 (1 987), describes 8- hyd roxy- 1 -met hyl-2-( di-n-propylami no) tetral ins. These compounds were 5-HT receptor agonists.

McDermed, et al., (J. Med. Chem., 18, 362 (1 975)) deecribes 5,6- dihydroxy-2-aminotetralins. In addition, the 5,8- and 7,8 di- substituted compounds are also disclosed. The amine can be a mono- or di-substituted with simple alkyl groups, benzyl groups, alkylalkoxy groups or the amine can be a 5- or 6-membered hydrocarbon or heterocyclic amine. These compounds are indicated

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to have dopaminergic properties although certain compounds are reported to be inactive.

McDermed, et al., (J. Med. Chem., 1 9, 547 (1 976)) describes 5-, 6-, or 7-hydroxy-2-dipropylaminotetralins. These compounds are described as dopaminergic compounds.

Rusterholz, et al., (J. Med. Chem., 1 9, 99 (1 976)) describes 5,8- disubstituted-2-aminotetralins with the amine being substituted with hydrogen, methyl, or cyanopropyl groups. Some of these compounds are potent prolactin inhibitors and believed to be DA agonists.

Ames, et al., (J. Chem. Soc. 2636 (1 965)) describes the preparation of a large number of compounds, where the aromatic ring is substituted by methoxy, ethoxy, n- or iso-propoxy, or n-, sec- or tert-butoxy group in the 5 or 8 position and the amine is substituted by hydrogen or alkyl groups having 1 -4 carbon atoms. The compounds are indicated to be prepared for pharmacological testing. However, no utility or pharmacological activity is yet known for the compounds just mentioned.

The German patent Number DE-AI-2 803 582 describes 2- aminotetralins where the aromatic ring is substituted on the 5,6,7 or 8 position with the group R1 , where R1 is hydrogen, alkanoyl having 1 to 20 carbon atoms or a group -CO-(CH2 )n~R7 , n is a number 0 to 5, R7 is a phenyl group with substituents as defined further, R2 is hydrogen, hydroxy, halogen or alkylsulfonylamino, R3

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is hydrogen, R4 is hydrogen, CH2OH, CH2O-CO-R8 or CH2-0-CO- (CH2)n-R7 with further definition and R5 and R6 are hydrogen, alkyl or aryl or aralkyl groups further defined or R5 and R6 are together an alkylene with 4 to 6 carbon atoms. The compounds are disclosed as having pharmacodynamic activity in particular a stimulating effect on alpha- and beta-adrenoceptors and DA receptors. Among the compounds described are compounds having the group R10 in 8 position and having R2 or R4 other than hydrogen.

The UK Patent Number 1 ,377,356 describes 2-aminotetralins where the aromatic ring is substituted on the 5, 6, 7 or 8 position by R1 , where Ri is hydrogen or methyl, the aliphatic ring is substituted by R2, where R2 is alkyl having 1-6 carbon atoms, and the amine is substituted by R3 , where R3 is hydrogen or alkyl having 1 -6 carbon atoms are described. Such compounds are stated to possess analgesic activity. 1 , 1 -Dimethyl-2-(N,N- dimethylamino)-7-hydroxytetralin is mentioned as one example of a compound covered by the patent. This compound is also described in Chem. Ab., 79: 146294b as having analgesic and intestinal movement accelerating actions.

J. Pharm. Sci., 67, 880-82 (1 978) describes the compound 1 - methyl-2-(cyclopropylamino)-5-methoxytetralin and indicates the compound possess local anesthetic activity.

Liu, Y. et al., "C8-Substituted derivatives of 2-

(dipropylamino)tetrlain: Palladium-catalyzed synthesis and

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interaction with 5-HT1 A receptors" have published the triflate derivative of 8-OH-DPAT in Bioorganic & Medicinal Chemistry Letters, 1991 , 1 , 257-262:

This compound (the racemate has the RN [1 34394-05-1 ]) was shown to bind to 5-HT1 A receptors with 14 nM affinity. No further pharmacology on this compound has been published as yet. This N,N-di-propylated triflate derivative falls outside the present claim (see below). The same compound appears also in a patent from Anden et al. (EP 399982 Al , November 28, 1990, with priority day May 26, 1989). In addition, the same compound and its enantiomers appear in Liu et al., J. Med. Chem. 1993, 36, 4221-29.

In a patent application from Lin et al. (PCT Int. Pat. Appl., WO 9206967 Al , April 30, 1 992, with priority day October 12, 1990) the enantiomers of the 8-triflated DPAT are used as intermediates to produce the enantiomers of 8-CN-DPAT. These 8-TfO-DPAT enantiomers have the following RN numbers: R-(+)-8-TfO-DPAT

[1 36376-13-1 ] and S-(-)-8-TfO-DPAT [136376-09-5]. Some of the mono-alkylated compounds of this patent application are within the general claim of the Lin et al., WO 9206967 A1 application, but no example of a N-mono-substituted analogue is given in their patent application. In addition, their claim is limited to triflates,

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CF3 -SO2-O-, and does not include other alkyl or aryl sulphone esters, like the R2 -Sθ 2 -0-derivatives of this application. Surprisingly, we found that the N-mono-alkylated sulphone ester analogues of this patent application are more potent and have better biological stability, as measured by their high oral bioavailability, than their corresponding N,N-dialkylated analogues.

The following documents are dealing with biologically active 3- aminochromanes:

Kidd, E. J.; Haj-Dahmane, S.; Jolas, T.; Lanfumay, L.; Fattaccini, C- M.; Guardiola-Lemaitre, B.; Gozlan, H.; Hamon, M., "New Methoxy- Chroman Derivatives, 4 [ N-(5-Methoxy-Chroman-3-yl)-N- propylamino]Butyl-Azaspiro-(4,5)-Decane-7,9-Dione[(±)-S2024 4] and Its Enantiomers, (+)-S20499 and (-)-S20500, with Potent Agonist Properties at Central 5-Hydroxytryptamineι A Receptors.", J. Pharmacol. Exp. Ther., 264(2), 863-72, 1993.

Guillaumet, G. ; Guardiola, B., "Preparation of 3- [(imidoalkyl)amino]chromans and analogs as 5-HT1 A receptor agonists.", EP 452204 A1 91 1016, PRAI 900409.

Cossery, J. M.; Perdicakis, C; Coudert, G.; Guillaumet, G.; Pichat, L., "Oxygen isosteres of hydroxy(dipropylamino)tetralins. Syntheses of racemic monomethoxy- and monohydroxy-3-[di[ 3 H- propyl]amino]chromans, new radioligands for 5-HT1 A and D2- receptor-site labeling.", J. Labelled Compd. Radiopharm., 25(8), 833-54, 1 988. SUBSTITUTE SHEET

Hall, H. 0.; Johansson, L. G.; Thorberg, S.-0., "5-Hydroxy3- aminochroman compounds, processes for their preparation, pharmaceutical compositions containing them and methods of treatment therewith.", WO 8804654 Al 880630, PRAI 86121 9.

Thorberg, S.-O.; Hall, H.; Akesson, C; Svensson, K.; Nilsson, J. L. G., "Aminochromans: potent agonists at central dopamine and serotonin receptors.", Acta Pharm. Suec, 24(4), 169-82, 1987.

During the time from submission of the original application (9301732-5, May 18, 1993) to the Swedish Patent Office (Patent och Registreringsverket), the following documents, relevant to this PCT application, have appeared:

Svensson, Wikstrom, Carlsson, Boije, Waters, Sonesson, Stjemlδf, Andersson, Hansson, Patent: WO 9218475, PRAI: US 686799, Filing date: 26 Mars, 1992.

Sonesson et al., J. Med. Chem., 1993, 36(21 ), 3188-96.

Sonesson et al., J. Med. Chem., 1993, 36(22), 3409-16.

Haadsma-Svensson et al. Bioorg. & Med. Chem. Lett., 1 994, 4(5), 689-94.

These documents confirm the statement in this application that aromatic sulfonates, in particular triflates, have beneficial

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pharmacokinetic properties. In addition, they confirm that aromatic sulfonates have interesting pharmacodynamic properties.

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Summary of the Invention

This invention is related to novel 2-aminotetralin compounds of Formula 1 :

Formula 1

or pharmaceutically acceptable acid addition salts thereof, wherein Ri is H, (Ci -Cs) alkyl, alkenyl, alkynyl, cyclopropylalkyl or (C-| -C8) haloalkyl; X is CH2, 0 or S; R2-SO2-O is in position 5, 6, 7 or 8 and R2 is CF3, CF2CF3, C1 -C8 alkyl, substituted aryl (e.g. 4- toluyl); R3 is H, CH3 or CH2CH3. When R3 is CH3 or CH2CH3, it is always in a cis-relationship with respect to the 2-amine substituent.

The compounds of this invention possess selective 5-HT1 A or 5- HT1 D pharmacological properties and are useful in treating hypertension, coughs (antitussive) and central nervous system disorders including depression symptoms, anxiety symptoms, panic attacks, obsessive-compulsive disturbances, senile dementia, emotional disturbances related to dementia disorders, and disturbances of sexual functions. The compounds of this invention

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are also useful to alleviate aggressive behavior, confusional delirious states and impotence. In addition, the dopaminergic compounds of this invention are useful for treating disturbances in the CNS such as parkinsonism, schizophrenia, MBD and mano- depressive illness.

In a preferred embodiment, the invention is directed to compounds of Formula 1 ; wherein Ri is C1 -C3, X is CH2 and R2 is CH3 or CF3 and R3 is H, CH3 or CH2CH3. A more preferred embodiment are compounds of Formula 1 ; wherein Ri is n-Pr, X is CH2 and R2 i s CH3 or CF3 and R3 is H, CH3 or CH2CH3. An even more preferred embodiment are compounds of Formula 1 ; wherein Ri is n-Pr, X is CH2 and R2 is CF3 and R3 is H. The most preferred positions for the aromatic substituent are 5 and 7 for DA effects and 8 for 5-HT effects.

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 the 5-HT1 A or 5-HT1 D receptors in mammals including man. A further object of this invention is to provide compounds having an effect on the subclasses of DA receptors known as the D2 and D3 receptors.

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

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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 (C1 -C8) 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 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, isohexyl, 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, I- 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. CycloalkyI refers to a radical of a saturated cyclic hydrocarbon such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl. "Halogen" means fluorine, chlorine, bromine or iodide, preferably fluorine. __

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It will be apparent to those skilled in the art that compounds of this invention may contain chiral centers. 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 two asymmetric carbon atoms in the aliphatic ring moiety, including the ring carbon atoms adjacent to the nitrogen atom. 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, maleic 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

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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.

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, preferentially 0.5-500 mg, and 0.05 mg to about 100 mg for parenteral application, preferentially 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.

The compounds of Formula 1 of this invention, where Ri is Ci -C3 alkyl, 2-SO2-O is in the 5, 6 or the 7 position, where R2 is CH3, CF3 or CF3CF2 and R3 is H, are DA D2 and/or D3 receptor agonists. The compounds of Formula 1 of this invention, where Ri is C1 -C3 alkyl, R2-SO2-O is in the 5 or the 7 position, where R2 is CH3, CF3 or CF3CF2 and R3 is cis-CH3, are DA D2 and/or D3 receptor antagonists.

The compounds of Formula 1 of this invention, where Ri is C1 -C3 alkyl, R -SO2-O is in the 8 position, where R2 is CH3, CF3 or

CF3CF2 and R3 is H or cis-CH3, are very selective 5-HT1 A and or

5-HT1 D receptor ligands (agonist and/or antagonists), having little or no dopaminergic activity. These compounds are particularly

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effective hypotensive, anxiolytic and antidepressant agents. Other uses for these compounds include panic attacks, obsessive- compulsive disturbances, and senile dementia, particularly the emotional disturbances seen in dementia disorders. In addition, central 5-HT receptor activation is believed to be involved in mediating sexual behavior. These compounds would be useful to stimulate sexual activity and to alleviate impotence.

The compounds of this invention with effects of DA receptors are particularly effective in treating psychoses, mano-depressive illness and parkinsonism. The DA autoreceptor antagonists of this invention are stimulants and can be useful in treating parkinsonism, MBD and depression, especially in elderly people.

The compounds of this invention also have high oral potency and a long duration of action, which is different from the 2- aminotetralins known to date (see the references given above). Both these features are beneficial to effective clinical treatment.

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

Antagonism of the reserpine-induced "neuroleptic syndrome" in the rat (gross behavior'..

Depletion of the monoamine stores with reserpine brings about a

"neuroleptic syndrome" characterized by hypomotiiity, catalepsy,

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muscle rigidity, hunch-backed posture, as well as a number of other central and peripheral signs of monoamine depletion. The whole or parts of this syndrome can be reversed by the administration of drugs that stimulate DA or 5-HT receptors directly or indirectly.

Stimulation of the DA receptors, with apomorphine for example, gives initially rise to yawning (a D3 effect) and later to both locomotion and stereotyped behavior such as sniffing, gnawing and jumping. On the other hand, stimulation of the 5-HT receptors, with 5-hydroxytryptophan (5-HTP) combined with MAO-inhibitors for example, gives rise to a very different behavior, "the 5-HT syndrome". The animals lie flat on the cage floor exhibiting forward movements with extended forepaws padding, "piano- playing," and abducted hindlegs, occasionally with some tremor in the forebody and with Straub tail (stiff tail erection).

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In vivo determination of rat brain tyrosine and tryptophan hvdroxvlation after reserpine ^p retreatment (biochemically monitored DA and 5-HT receptor activity).

5 The compounds under evaluation were tested biochemically for central DA and 5-HT receptor (pre- and/or postsynaptic) stimulating activity. The concept of this biochemical screening method is that a DA or 5-HT-receptor agonist will stimulate the receptor and through regulatory feedback systems effect a decline

10 in tyrosine or tryptophan hydroxylating activity, respectively, and a subsequent reduction in the synthesis rate for DA and 5-HT in the presynaptic neuron. DOPAC and 5-HTP formation, as determined after in vivo inhibition of the aromatic L-amino acid decarboxylase with NSD 101 5 (3-hydroxybenzylhydrazine hydrochloride) are taken

1 5 as indirect measures of DA and 5-HT synthesis rates, respectively, as described by Wikstrδm et al., J. Med. Chem. 27, 1030, 1984.

Analogous conditions probably exist also for central NA-neurons. Effects on the DA formation in the NA-predominated hemispheral 20 parts (mainly cortex) may thus be considered to reflect NA- receptor mediated changes.

Anti-anxietv models in vivo: 1 ) Elevated Plus Maze and 2 Pefeηsive B ryiηg,

25

1 ) The rats were placed in the middle of an elevated (1 m above the floor) plus maze having two open arms and two closed arms. The total number of entries in either type of arm was scored in drug

treated and saline treated (control) rats. More entries in the open arms is considered to be an expression of diminished anxiety.

2) The rats were placed in a cage equipped with en electrified rod. After the first contact with the rod the rat starts to bury the rod in order to avoid electric chock. The more anxious, the more time the rat devotes to burying and vice versa.

Experimental procedures

Syntheses

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

L. E. Arvidsson, U. Hacksell, J. L. G. Nilsson, S. Hjorth, A. Carlsson, P. Lindberg, D. Sanchez and H. Wikstrόm, 8-Hydroxy-2- (dipropylamino)tetralin, a new centrally acting 5- hydroxytryptamine receptor agonist, J. Med. Chem., 1981 , 24, 921 - 3, Abstract: Four monophenolic tetralin derivs. I were synthesized and tested in rats for dopamine (DA) and 5-hydroxytryptamine (5- HT) receptor-stimulating activity. I (8-OH-DPAT) [761 35-31 -4] was a potent and selective 5-HT receptor agonist devoid of DA receptor stimulating activity. (+)-l (8-OH-DPAT) [78095-1 9-9] was a more active 5-HT receptor agonist than (-)-l (8-OH-DPAT)

[78095-20-2]. I (5-OH-DPAT) [71 787-83-2], I (6-OH-DPAT)

[761 35-29-0], and I (7-OH-DPAT) [761 35-30-3] were potent DA

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receptor agonists and lacked 5-HT receptor stimulating activity. Structure-activity relations are discussed.).

L. E. Arvidsson, U. Hacksell, A. Johansson, J. L. G. Nilsson, P. Lindberg, D. Sanchez, H. Wikstrόm, K. Svensson, S. Hjorth and A. Carlsson, 8-Hydroxy-2-(alkylamino)tetralins and related compounds as central 5-hydroxytryptamine receptor agonists, J. Med. Chem., 1984, 27, 45-51, Abstract: The title compds. I (RI = H, OH, or OMe; R2 = OH or OMe; R3 = H or C1-4 alkyl; R4 = C1-4 alkyl, etc.; R3R4 = (CH2)5; HBr or HCl salts) were prepd. and tested as dopamine and 5-hydroxytryptamine-receptor agonists in reserpinized rats. (R)-(+)-8-Hydroxy-2-(dipropylamino)tetralin- HCI[(R)-(+)-l; RI = H, R2 = OH, R3 = R4 = Pr.HCI] showed the highest activity. Structure-activity relations are discussed.

L. E. Arvidsson, A. M. Johansson, U. Hacksell, J. L. G. Nilsson, K. Svensson, S. Hjorth, T. Magnusson, A. Carlsson, B. Andersson and H. Wikstrom, ( + )-cis-8-Hydroxy-1 -methyl-2-(di-n- propylamino)tetralin: a potent and highly stereoselective 5- hydroxytryptamine receptor agonist, J. Med. Chem., 1987, 30, 2105-9, Abstract: C(1)-methylated derivs. of the potent 5- hydroxytryptamine (5-HT) receptor agonist 8-hydroxy-2- (dipropylamino)tetralin (I) were synthesized and tested for central 5-HT and dopamine receptor activity by use of a biochem. test method in rats. cis-8-Hydroxy-1-methyl-2-(dipropylamino)tetralin

(II) was found to be a 5-HT receptor agonist. The (+)-enantiomer of II had a potency equal to that of I, whereas (-)-ll and the trans isomer (.+-.)-lll were inactive.

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H. Wikstrόm, B. Andersson, D. Sanchez, P. Lindberg, L. E. Arvidsson, A. M. Johansson, J. L. G. Nilsson, K. Svensson, S. Hjorth and A. Carlsson, Resolved monophenolic 2-aminotetralins and 1 ,2,3,4,4a,5,6,10b-octahydrobenzo[f]quinolines: structural and stereochemical considerations for centrally acting pre- and postsynaptic dopamine-receptor agonists, J. Med. Chem., 1985, 28, 21 5-25, The resolved title compds. 2-(dipropylamino)-5-hydroxy- and -7-hydroxytetralins I (R = 5- or 7-OH) and cis- and trans- propylbenzo[f]quinolinols (II) prepd. by demethylation of the appropriate methoxy compd. were evaluated for a detailed structure-activity relationship of their pre- and postsynaptic dopamine receptor-agonist activity. Male rats were used in the biochem. and motor activity expts. (S)-2-(Dipropylamino)-5- hydroxytetralin (I; R = 5-OH-DPAT) [68643-08-3] and (R)-2- (dipropylamino-7-hydroxytetralin (I; R = 7-OH-DPAT) [82730-72- 1 ] were the most active compds.

Other relevant references (journals and patent applications) are:

Al, N. M.; Reynaud, D.; Podona, T.; Ou, L.; Perdicakis, C; Coudert, G.; Guillaumet, G.; Pichat, L.; Gharib, A.; Sarda, N. Methoxy and hydroxy derivatives of 3 ,4-dihydro-3-(di-n-propylamino)- 2H- 1 - benzopyrans: new synthesis and dopaminergic activityD. Eur. J. Med. Chem. 1991 , 26,

Chen, J.; Hutchison, A. J. EP 267878 A1 88C51 8: N9-Cyclopentyl- substituted adenine derivatives, procedure for their preparation,

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pharmaceutical compositions containing them, and their use as adenosine receptor agonistsD. Ep 1988,

Chen, J.; Hutchison, A. J. US 4954504 A 900904: Preparation of n9-cyclopentyl-substituted adenine derivatives having adenosine- 2 receptor stimulating activityD. Us 1990,

Ciba, G. A.; G JP 62059273 A2 870314 Showa: Preparation of 3- aminodihydro[ 1 ]benzopyran and benzothiopyran derivatives as serotoninergic agonistsD. Jp 1987,

Cossery, J. M.; Gozlan, H.; Spampinato, U.; Perdicakis, C; Guillaumet, G.; Pichat, L.; Hamon, M. The selective labelling of central 5-HT1 A receptor binding sites by [3H]5-methoxy-3-(di-n- propylamino)chromanD. Eur. J. Pharmacol. 1987, 140, 143-55.

Cossery, J. M.; Perdicakis, C; Coudert, G.; Guillaumet, G.; Pichat, L. Oxygen isosteres of hydroxy(dipropylamino)tetralins. Syntheses of racemic monomethoxy- and monohydroxy-3 -[di [ 3 H- propyl]amino]chromans: new radioligands for 5-HT1 A- and D2- receptor-site labelingD. J. Labelled Compd. Radiopharm. 1 988, 25, 833-54.

Guillaumet, G.; Guardiola, B. EP 452204 A1 91 1016: Preparation of 3-[(imidoalkyl)amino]chromans and analogs as 5-HT1 A receptor agonistsD. Ep 1991 ,

SUBSTITUTE SHEET

Guillaumet, G.; Coudert, G.; Podona, T.; Guardiola, L. B.; Renard, P.; Adam, G.; Caignard, D. H. EP 571 243 Al 931 124: Preparation of (aminoalkoxy)thiochromans as psychotropicsD. Eur. Pat. Appl. 1993,

Hall, H. 0.; Johansson, L. G.; Thorberg, S. 0. WO 8804654 Al 880630: 5-hydroxy-3-aminochroman compounds, processes for their preparation, pharmaceutical compositions containing them and methods of treatment therewithD. Pet Int. Appl. 1988,

Hammarberg, E. M.; Johansson, L. G.; Ross, S. B.; Thorberg, S. 0. EP 538222 Al 930421 : 3-(N-isopropyl-N-propylamino)-5-(N- isopropyl)carbamoylchroman as a selective 5-hydroxytryptamine receptor stimulantQ. Eur. Pat. Appl. 1993,

Hutchison, A. J. EP 277917 A2 880810: Preparation of adenosine- 5'-carboxamide derivatives as adenosine-2 receptor agonists, antipsychotics, and antihypertensives and pharmaceutical compositions containing themD. Ep 1988,

Hutchison, A. J. EP 280269 Al 880831 : Preparation of 3- aminodihydro-2H-[ 1 ]benzopyrans and -thiopyrans as neurotransmitter agonistsD. Ep 1988,

Hutchison, A. J. US 4968697 A 901 106: Preparation of 2- substituted adenosine-5'-carboxamides as antihypertensive agentsD. Us 1990,

SUBSTITUTE SHEET

Larsson, L. G.; Noreen, R.; Renyi, L. A.; Ross, S. B.; Sohn, D. D.; Svensson, B. E.;

Thorberg, S. 0. WO 91 09853 Al 91 071 1 : Preparation of 3- (alkylamino)chroman and -thiochroman derivatives and pharmacological activity thereofD. Pet Int. Appl. 1991 ,

Thorberg, S. 0.; Hall, H.; Aakesson, C; Svensson, K.; Nilsson, J. L. G. Aminochromans: potent agonists at central dopamine and serotonin receptorsD. Ada Pharm. Suec. 1987, 24, 169-82.

SUBSTITUTE SHEET

Experimental Section

1 H and 1 3C NMR spectra were recorded at 200 and 50.3 MHz, respectively, on a Varian Gemini 200 spectrometer. CDCI3 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. The chemical ionization (CI) mass spectra were obtained on a Finnegan 3300 system or a UNICAM Automass 1 50 GC/MS system, equipped with a UNICAM GC, 61 0 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 monomethoxylated aminotetralins were prepared according to the literature procedures. Chemicals used were commercially available (Aldrich and/or Sigma) and were used without further purification. R- and S-2-Chlocyphos were obtained from Syncom B.V, The Netherlands.

Parts of the synthetic methods for forming the sulphone esters from the corresponding phenols are exemplified in Scheme 1 :

SUBSTITUTE SHEET

Scheme 1

Suitable starting materials (either the phenols or the OMe- analooues. which can be dealkvlated to the corresponding phenols in refluxing (2 h) 48% HBr or in CH^CI^ with BBr for this invention are known:

(±)-8-OH-2-(N-n-propylamino)tetralin; RN [105578-37-8]

SUBSTITUTE SHEET

(±)-8-OMe-2-(N-n-propylamino)tetralin; RN [87395-00-4] or

[3902-22-5]

R-(+)-8-OMe-2-(N-n-propylamino)tetralin; RN [81 185-23-1 ]

S-(-)-8-OMe-2-(N-n-propylamino)tetralin; RN [1 33267-79-5] (±)-7-OMe-2-(N-n-propylamino)tetralin; RN [1 36559-47-2] and

[148258-42-8]

(+)-7-OMe-2-(N-n-propylamino)tetralin; see Wikstrόm et al. J. Med.

Chem., 28, 215-25, 1985.

(-)-7-OMe-2-(N-n-propylamino)tetralin; ; see Wikstrδm et al. J. Med. Chem., 28, 215-25, 1985.

(±)-6-OMe-2-(N-n-propylamino)tetralin; RN [82763-23-3]

(±)-5-OMe-2-(N-n-propylamino)tetralin; RN [3899-07-8] and

[78598-91 -1 ]

R-(+)-5-OMe-2-(N-n-propylamino)tetralin; RN [101403-25-2] S-(-)-5-OMe-2-(N-n-propylamino)tetralin; RN [101403-24-1 ]

(±)-5-OH-2-(N-n-propylamino)tetralin; RN [78598-89-7] or

[78950-82-0]

R-(+)-5-OH-2-(N-n-propylamino)tetralin; RN [101470-24-0]

S-(-)-5-OH-2-(N-n-propylamino)tetralin; RN [101470-23-9] (±)-2H-Benzopyran-3-amine, 3,4-dihydro-5-methoxy-N-propyl-, or

5-Methoxy-3-(N-propylamino)chromane; RN [1 1 9755-71 -4] and RN

[1 12904-79-7]

(±)-2H-Benzopyran-3-amine, 3,4-dihydro-5-methoxy-, or 5-

Methoxy-3-aminochromane; RN [1 19755-62-3] and [1 10927-03-2] R-2H-Benzopyran-3-amine, 3,4-dihydro-5-methoxy-, or R-5-

Methoxy-3-aminochromane; RN [1 17444-30-1 ]

S-2H-Benzopyran-3-amine, 3,4-dihydro-5-methoxy-, or S-5-

Methoxy-3-aminochromane; RN [1 17422-50-1 ]

SUBSTITUTE SHEET

(-)-2H-Benzopyran-3-amine, 3,4-dihydro-5-methoxy-, or (-)-5- Methoxy-3-aminochromane; RN [1 17422-48-7]

2H-1 -Benzothiopyran-5-ol, 3,4-dihydro-3-(propylamino)-, or 5- hydroxy-3-(N-propylamino)-dihydro-2H-[ 1 ]benzothiopyran; RN [1 1 2904-80-0]

2H-1 -Benzothiopyran-5-methoxy, 3,4-dihydro-3-amino-, or 5- methoxy-3-dihydro-2H-[1 ]benzothiopyran; RN [109140-19-4]

Resolution 8-methoxy-2-(N-n-propylamino)tetralin

Resolution oj 8-QMe-2-(N-n-prQpylamirtQ)tetralin (first desc ptipn),

8-OMe-2-(N-n-propylamino)tetralin (1 1 .1 1 g; 50.7 mmol) was dissolved in EtOH (1 00 mL) and (+)-R-2-Chlosyphos (14.00 g; 50.7 mmol) was dissolved in EtOH (100 mL) and the two solutions were merged. The solvent was evaporated under reduced pressure. The residue was refluxed in acetone (300 mL) and white crystals were filtered after cooling. This costitutes a purification process and not an actual resolution crystallization. The white crystals (5.0 g) were recrystallized two times from refluxing i-PrOH and gave crystals (3.7 g) with an [α]D ^{2 1} = +60.1 ° (c=1.0 in MeOH). These crystals (3.7 g) were basified with 1 0% KOH and extracted with

CH2CI2 , yielding a colorless oil (1 .6 g; 1 4%) with an [α]D ²¹ = +76.4° (c=1 .0 in MeOH), the R-enantiomer according to the literature.

SUBSTITUTE SHEET

The residueal filtrate from the acetone crystallization (above) (1 1 .24 g) was basified with 10% KOH and extracted with CH2CI2, yielding a colorless oil (4.8 g), which was dissolved in EtOH (100 mL) and mixed with (-)-S-2-Chlosyphos (6.0 g; 21 .7 mmol) dissolved in EtOH (100 mL). The solvent was evaporated under reduced pressure. The residue was refluxed in acetone (200 mL) and white crystals were filtered after cooling. This costitutes a purification process and not an actual resolution crystallization. The white crystals were recrystallized two times from refluxing i-PrOH and gave crystals (4.0 g) with an [α]D ²¹ = -61.2° (c=1.0 in MeOH). These crystals (3.9 g) were basified with 10% KOH and extracted with CH2CI2, yielding a colorless oil (1.7 g; 1 5%) with an [α]D ²¹ = -77.6° (c=1.0 in MeOH), the S-enantiomer according to the literature.

R-(+V8-OH-2-(N-n-propvlamino)tetralinxHBr

R-(+)-8-OMe-2-(N-n-propylamino)tetralin (1 .7 g; 6.8 mmol) was refluxed for 2 h in 48% HBr (50 mL) under N2(g). After cooling to room temprature the excess HBr was evaporated under reduced pressure yielding a white solid, which was dried in a desiccator, yielding 1.88 g (97%; [α]D ²¹ = +63.5° (c=1.0 in MeOH)), which was used in the triflation step without further purification.

S-(-)-8-OH-2-(N-n-propylamino)tetralinxHBr

S-(-)-8-OMe-2-(N-n-propylamino)tetralin (1 .9 g; 7.5 mmol) was refluxed for 2 h in 48% HBr (75 mL) under N2(g). After cooling to

SUBSTITUTE SHEET

room temprature the excess HBr was evaporated under reduced pressure yielding a white solid, which was dried in a desiccator, yielding 2.1 7 g (100%; [α]D ²¹ = -64.5° (c=1.0 in MeOH)), which was used in the triflation step without further purification.

R-( ₊ V8-OSθ£CF^-2-(N-n-propvlamino)tetralinxHCI

R-(+)-8-OH-2-(N-n-propylamino)tetralinxHBr (0.92 g; 3.2 mmol) was dissolved in CH2CI2 (50 mL) and 10% NaOH (50 mL) was added. Thereafter was added QHS04 (Q=NBu4 ⁺ ; 109 mg, 10 mol %) and PhN(S02CF3)2 (1 .26 g; 3.5 mmol) (see Scheme 1 above) and the reaction mixture was stirred vigurously to give an optimal phase contact. After 3 h the organic phase was separated and the water phase was extracted once with another portion of CH2CI2 (50 mL). The organic phases were pooled and the solvent was evaporated and the residue was redissolved in ether (50 mL). The ether phase was washed with water and extracted with 5 % HCl. The ether phase was separated and the acidic water phase was basified with Na2C03(s) and the product was extracted over in ether (3x100 mL). The ether phases were pooled, dried (Na2S04) and the solvent was evaporated under reduced pressure to give a colorless oil (0.43 g; 40%). This oil (0.33 g) was dissolved in ether and converted to its HCl salt with ethereal HCl, yielding white crystals (347 mg; 38%) melting at 235-238 °C and having [a ]D ²¹ = +61.5° (c=1.0 in MeOH).

S-(-V8-OSO?CF;3-?-(N-n-nropylaminoHetralinxHCI

SUBSTITUTE SHEET

S-(-)-8-OH-2-(N-n-propylamino)tetralinxHBr (0.88 g; 3.1 mmol) was dissolved in CH2CI2 (100 mL) and 10% NaOH (50 mL) was added. Thereafter was added HSO4 (107 mg, 1 0 mol %) and PhN(S02CF3)2 (1.25 g; 3.5 mmol) (see Scheme 1 above) and the reaction mixture was stirred vigurously to give an optimal phase contact. After 2 dr 's the organic phase was separated and the water phase was extracted once with another portion of CH2CI2 (50 mL). The organic phases were pooled and the solvent was evaporated and the residue was redissolved in ether (50 mL). The ether phase was washed with water and extracted with 5 % HCl. The ether phase was separated and the acidic water phase was basified with Na2Cθ3(s) and the product was extracted over in ether (3x100 mL). The ether phases were pooled, dried (Na2Sθ4) and the solvent was evaporated under reduced pressure to give a colorless oil (1.0 g; 97%). This oil (0.91 g) was dissolved in ether and converted to its HCl salt with ethereal HCl, yielding white crystals (760 mg; 73%) melting at 235-238 °C and having [α]D ²¹ = -61 .6° (c=1.0 in MeOH).

SUBSTITUTE SHEET

The synthetic sequence described above has been rewritten and more experimental details and physical data have been added according to the following:

Resolution of 8-Methoxy-2-(n-propylamino ^' )tetralin (second description).

A mixture of racemic amine (1 1 .1 g, 50.7 mmol) and R-(+)-2- chlocyphos (14.0 g, 50.7 mmol) in abs. ethanol (200 mL) was refluxed until all material was dissolved after which the solvent was removed in vacuo giving an off-white solid. The salt (24.1 g, 48.7 mmol) was recrystallized from 2-propanol yielding 5.1 6 g (10.42 mmol, 21 %) of white crystals with [α]D ²¹ = +53.1. A second recrystallization gave 3.74 g (7.56 mmol, 16%) salt with [α]D ²¹ = +60.1 (c = 1.0 in MeOH). This salt (3.65 g, 7.37 mmol) was converted to the free base by stirring in 1 0% KOH (50 mL), extraction with ether and drying over Na2S04. Evaporation of the solvent in vacuo yielded R-( + )-8-methoxy-2-( n- propylamino)tetralin (1.58 g, 1 5%) as a colorless oil with [α]D ¹ = +76.4 (c * 1.0 in MeOH) (litl [ ]D ²² = +78.3 (c=1.05)). The residual salt (1 1 .2 g, 22.7 mmol) was converted to the free base described as above using 10% KOH (100 mL). Repeating the above procedure with the enriched (-)-enantiomer of 8-methoxy-2-(n- propylamino)tetralin (4.57 g, 21 ,7 mmol) with S-(-)-chlocyphos ( 5.99 g, 21 .7 mmol) gave S-(-)-8-methoxy-2-(n- propylamino)tetralin (1.65 g, 15%) as a colorless oil with [α]D ²¹ = -77.6 (c = 1.0 in MeOH) (litl [ ]D ²² = -77.0 (c=1.03)).

SUBSTITUTE SHEET

R-(+)-8-Hvdroxy-2-(n-propylamino)tetralin hvdrobromide.

R-(+)-8-Methoxy-2-(n-propylamino)tetralinxHCI salt (1 .74 g, 6.82 mmol) was refluxed in 48% HBr (50 mL, freshly distilled) for 2 h under N2-atmosphere. The reaction mixture was allowed to cool to room temperature and evaporated to dryness giving 1 .88 g (97%) of the product as a pale-brown solid, of which 445 mg was recrystallized from ethanol/ether for purification (374 mg, 78%): mp 283-286 °C; IR (KBr) 3275 cm-1 ; I H NMR (CD30D) 1.07 (t, J = 7.69, 3H), 1.70-1.91 (m, 3H), 2.33 (m, I H), 2.60 (dd, Jl = 10.25, J2 = 1 6.23, I H), 2.91 (m, 2H), 3.08 (m, 2H), 3.26-3.37 (m, 1 H), 3.43- 3.58 (m, 1 H), 6.61 (d, J = 7.7, 1 H) 6.62 (d, J = 8.12, 1 H) 6.97 (dd, Jl = 7.69, J2 = 8.12, 1 H); 1 3C NMR (CD30D) 1 1.0, 20.7, 26.6, 27.2, 28.3, 47.5, 55.8, 1 12.6, 120.0, 120.3, 127.8, 136.9, 1 56.0; MS (CI with NH3) m/e 206 (M+1 ); Anal. Calcd for C13H19NOxHBr: C, H, N; [α]D ²¹ = +63.5 (HBr) (c = 1.0 in MeOH).

S-(-V8-Hvdroxv-2-(n-propvlamino')tetralin hvdrobromide.

Demethylation of S-(-)-8-methoxy-2-(n-propylamino)tetralin (1.92 g, 7.53 mmol) was performed according to procedure as described for R-(+)-8-methoxy-2-(n-propylamino)tetralin as above giving S-(-)-8-hydroxy-2-(n-propylamino)tetralinxHBr in a quantitative yield. Part of the salt (1 .06 g) was recrystallized from ethanol/ether yielding 0.80 g (76%) of off-white crystals: mp

273-277 °C; IR (KBr) 3275 cm-1 ; I H NMR (CD30D) 1.07 (t, J = 7.69,

3H), 1.70-1.91 (m, 3H), 2.33 (m, 1 H), 2.60 (dd, J1 = 10.25, J2 =

16.23, I H), 2.91 (m, 2H), 3.08 (m, 2H), 3.26-3.37 (m, I H), 3.43-3.58

SUBSTITUTE SHEET

(m, I H), 6.61 (d, J = 7.7, I H) 6.62 (d, J = 8.12, 1 H) 6.97 (dd, Jl = 7.69, J2 = 8.12, 1 H); 13C NMR (CD30D) 1 1 .1 , 20.7, 26.6, 27.2, 28.3,

47.5, 55.8, 1 12.6, 120.0, 120.3, 127.8, 136.9, 1 56.0; MS (CI with NH3) m/e 206 (M+1 ); Anal. Caicd for C13H19NO.HBr: C, H, N; [α]D ²¹ = -64.5 (HBr) (c = 1.0 in MeOH).

R-(+)-8-rr(Trifluoromethvhsulfonyl1oxy1-2-(n- propvlamino)tetralin.

A mixture of R-(+)-8-hydroxy-2-(n-propylamino)tetralinxHBr (200 mg, 0.70 mmol), N-phenyltrifluoromethanesulfonimide (376 mg, 1 .05 mmol) and tetrabutylammoniumhydrogensulfate (24 mg, 10 mol%) in dichloromethane (8 mL) and 10% NaOH (3 mL) was stirred at room temperature for 24 h. The reaction mixture was quenched with 5% HCl solution (v/v) until pHl , diluted with H20 (25mL) and washed with ether (50 mL). The ether layer was extracted with H20 and 5% HCl solution (20 mL). The combined aquous layers were basified with solid Na2C03 until pH 9, extracted with ether (3X30 mL) after which the organic phase was washed with brine and dried over Na2S04. Evaporation in vacuo yielded the product as a colorless oil, which was converted to the HCl salt and recrystallized from methanol/ether (177 mg, 68%): mp 238-240 °C (HCl); IR (KBr) 1217 cm-1 ; I H NMR 0.96 (t, J = 7.5, 3H), 1.35 (br s, NH), 1.55 (m, 2H), 1.63 (m, 1 H), 2.05 (m, 1 H), 2.53 (dd, Jl = 8.55, J2 = 16.24, I H), 2.69 (t, J = 7.5, 2H), 2.83-3.04 (m, 3H), 3.12 (dd, Jl = 4.71 , J2 = 16.24, 1 H), 7.05-7.18 (m, 3H); 13C NMR 1 1.8, 23.4, 27.8,

28.6, 30.7, 49.0, 52.5, 1 18.3, 1 18.6 (q, J = 321 , CF3), 126.7, 128.6,

128.7, 139.9, 148.4; MS (CI with NH3) m/e 338 (M+l ); Anal. Calcd

SUBSTITUTE SHEET

for C14Hl8N03SF3xHCI: C, H, N, F, S; [ ]D ²¹ = +61.5 (HCl) (c = 1.0 in MeOH).

S-(-V8-r[(Trifluoromethvnsulfonvlloxvl-2-(n- propylamino)tetralin.

Triflation of S-(-)-8-hydroxy-2-(n-propylamino)tetralinxHBr (880 mg, 3.08 mmol) was performed according to the procedure given for the synthesis of R-(+)-8-[[(trifluoromethyl)sulfonyl]oxy]-2-(n- propylamino)tetralin above giving an oil (1.0 g, 97%) after extractive workup. Conversion of this oil (0.91 g) to the HCl salt and subsequent recrystallization from methanol/ether gave 760 mg (73%) of white crystals: mp 235-238 °C (HCl); IR (KBr) 1217 cm-1; 1H NMR 0.95 (t, J = 7.7, 3H), 1.32 (br s, NH), 1.54 (m, 2H), 1.64 (m, 1H), 2.06 (m, IH), 2.53 (dd, J1 = 8.55, J2 = 16.24, IH), 2.69 (t, J = 7.69, 2H), 2.82-3.02 (m, 3H), 3.12 (dd, J1 = 4.7, J2 = 16.24, 1H), 7.04-7.20 (m, 3H); 13C NMR 11.7, 23.4, 27.7, 28.6, 30.7, 49.0, 52.5, 118.3, 118.6 (q, J = 321, CF3), 126.7, 128.6, 128.7, 139.9, 148.4; MS (CI with NH3) m/e 338 (M+l); Anal. Calcd for C14H18N03SF3.HCI: C, H, N, F, S; [α]D ²¹ = -61.6 (HCl) (c = 1.0 in MeOH).

Resolution 5- and 7-methoxy-2-( N-n- propylamino)tetralins. The amines have been resolved with the two enantiomers of 4-(2-chlorophenyl)-5,5-dimethyl-2-hydroxy- 1 ,3,2-dioxaphosphorane-2-oxide (Chlocyphos).

Resolution of 5-methoxy-2-(N-n-propvlamino) tetralin

SUBSTITUTE SHEET

A mixture of 6.23 g (28.4 mmol) racemic amine and 7.8 g (28.4 mmol) (+)-chlocyphos was dissolved in of ethanol (30 mL) and of H2θ (10 mL). The solution was allowed to cool to room temperature with stirring. After 6 h crystals were collected and washed, first with ethanol/ether and then with ether. The yield was 43% salt with [α]D ^{2 1} = -1.5 (c =1.0 in MeOH). The salt was stirred for 1 h with sodium hydroxide (1 g) in H2θ (50 mL) and then chloroform was added. The mixture was stirred for another 30 min. The layers were separated and the aqueous layer was extracted with chloroform. The chloroform layers were washed with water, dried over Na2S04 and evaporated to give an oil. Treatment with dry HCI- ether and recrystalliazation from ethanol/ether gave 3.1 g (43%) of a white salt melting at 279-283 °C and having an [α]D ²¹ = -65.2 (c=1 .0 in MeOH; lit -63.0). The filtrate was, after evaporation of the solvent, also treated with NaOH and gave impure amine. Treatment of this amine with (-)-chlocyphos afforded 2.5 g (35 %) optically pure (+)-amine [α]D ²¹ = +68.2 (c=1.0 in MeOH; lit +69.7). The e.e. was determined by 31 p-NMR ( 0.33 ppm) using the adduct of the acid chloride of chlocyphos.

Resolution of 7-methoxv-2-(N-n-propvlamino') tetralin

A mixture of 6.36 g (29 mmol) racemic 7-methoxy-2-(N-n- propylamino)tetralin (RN [1 36559-47-2]) and 8.0 g (29 mmol) of (-

)-chlocyphos was dissolved in of 2-propanol (450 mL). The solid was allowed to cool to room temperature with stirring. During the first hour optically pure seeding crystals were added. After 4

SUBSTITUTE SHEET

hours a white solid was filtered off. It displayed a rotation of [α]D ^{2 1} = +0.5 (c=1 .0 in MeOH). A second recrystallization from propanol-2 gave 6.0 g ( 42%) salt with [α]D ²¹ = +14.6 (c=1.0 in MeOH). This salt was stirred with sodium hydroxide in water (50 mL) and the aqueous layer was extracted with ethyl acetate. The organic layer was dried over MgSθ4. After filtration and evaporation of ethylacetate, a HCl salt was prepared. Recrystallization from ethanol/ether gave 2.5 g (33.7 %) [α]D ²¹ = +75.2 (c=1.0 in MeOH; lit. +70.2)

The filtrates from the two crystallizations were combined, concentrated, made alkaline and extracted with CH2CI2, dried over Na2Sθ4 and concentrated to give 4.1 g (> 50%) of the remaining 7- methoxy-2-(N-n-propylamino)tetralin after the first resolution above. This amine, (+)-chlocyphos (5 g, 1 8 mmol) and 2-propanol (200 mL) were heated to solution. The heating mantle was removed and the solution was allowed to cool to room temperature with stirring. After 6 h the salt was filtered off and washed with ether to give 6.4 g (12.9 mmol) with [α]D ²¹ = - 14.3 (c=1.0 in MeOH). This salt was converted into the HCl salt, and recrystallization from ethanol yielded 2.8 g (37.7%) S-(-)-7-methoxy-2-(N-n- propylamino)tetralin with an [α]D ² ^ = -75.1 (c=1.0 in MeOH; lit. -72.6).

(-)-5-Hvdroxy-2-(N-n-propylamino)tetralinxHBr

The hydrochloride salt of 5-methoxy-2-(N-n-propylamino)tetralin

(0.5 g, 1.96 mmol) was refluxed with 48% HBr (20 mL) under

SUBSTITUTE SHEET

nitrogen atmosphere for 3 h. Addition of H2θ gave a crystalline material, which on recrystallization from ethanol/ether yielded 350 mg (62.4%) product melting at 249-251 °C and with an [α]D ²¹ = -58.4 (c=0.76 in MeOH).

The procedure described above was also followed for the other enantiomer and the 7-methoxy isomers. The data for those reactions were:

(R)-(+)-5-OH-PATxHBr (72%)

(R)-(+)-5-OH-PATxHBr (72%)

(S)-(-)-7-OH-PATxHBr (61 %)

R-(+ 7-Hvdroxv-2-(n-propvlamino)tetralin hvdrobromide

Demethylation of R-(+)-7-methoxy-2-(n-propylamino)tetralin (see Wikstrom et al. J. Med. Chem., 28, 21 5-25, 1 985; 537 mg, 2.1 1 mmol) was performed according to procedure as described for R- (+)-8-methoxy-2-(n-propylamino)tetralin above, giving R-(+)-7- hydroxy-2-(n-propylamino)tetralin as a pale-brown solid. The title compound was purified by recrystallization from ethanol/ether (478 mg, 79%): mp 21 5-224 °C; IR (KBr) 331 5 cm-1 ; 1 H NMR (CD30D) 1.07 (t, J = 7.41 , 3H), 1.70-2.00 (m, 3H), 2.36 (m, 1 H),

2.91 -3.16 (m, 5H), 3.30-3.42 (m, 1 H), 3.51 -3.65 (m, I H), 7.13-7.33

(m, 3H); 13C NMR (CD30D) 1 1.0, 20.6, 26.4, 27.6, 32.5, 47.6, 54.8,

120.3, 122.5, 131.5, 136.0, 136.9, 148.9; MS (CI with NH3) m/e 206

SUBSTITUTE SHEET

(M+l ); Anal. Calcd for CI 3H1 9NO.HBr: C, H, N; [α]D ²¹ = +57.8 (HBr) (c = 1.0 in MeOH).

R-(+)-7-rr(Trifluoromethvnsulfonvl1oxv1-2-(n- propvlaminoketralin.

Triflation of R-(+)-7-hydroxy-2-(n-propylamino)tetralin (302 mg, 1.06 mmol) was performed according to the procedure given for the synthesis of R-(+)-8-[[(trifluoromethyl)sulfonyl]oxy]-2-(n- propylamino)tetralin above giving a colorless oil after extractive workup. Conversion to the HCl salt and subsequent recrystallization from methanol/ether gave 107 mg (34%) as a white crystalline material: mp 206-208 °C (HCl); IR (KBr) 121 1 cm-1 ; I H NMR (CD30D) 1.00 (t, J = 7.31 , 3H), 1.54-1.77 (m, 3H), 2.13-2.27 (m, I H), 2.64-3.23 (m, 7H), 7.06-7.26 (m, 3H); 13C NMR (CD30D) 1 1.6, 22.7, 28.1 , 28.5, 35.2, 48.9, 54.2, 1 19.6, 1 19.8 (q, J = 31 9, CF3), 122.3, 131.3, 137.7, 138.1 , 148.7; MS (CI with NH3) m/e 338 (M+1 ); Anal. Calcd for C14H18N03SF3.HCI: C, H, N; [α]D ²¹ = +50.6 (HCl) (c = 1.0 in MeOH).

S-(-V5-Hvdroxv-2-(n-propvlamino ^, .tetralin hvdrobromide

Demethylation of S-(-)-5-methoxy-2-(n-propylamino)tetralinxHCI

(61 7 mg, 2.42 mmol) was performed according to procedure as described for R-(+)-8-mβthoxy-2-(n-propylamino)tetralinxHCI above, giving the title compound as a pale-brown solid.

Recrystallization from ethanol/ether gave white crystals (312 mg,

45%): mp 249-251 °C; IR (KBr) 3327 cm-1 ; 1 H NMR (CD3OD) 1.07

SUBSTITUTE SHEET

(t, J = 7.45, 3H), 1.69-1 .92 (m, 3H), 2.30-2.43 (m, I H), 2.56-2.74 (m, I H), 2.82-3.35 (m, 5H), 3.41 -3.55 (m, 1 H), 6.63 (d, J = 7.69, 2H) 6.97 (t, J = 7.69, 1 H); 13C NMR (CD30D) 1 1.0, 20.7, 22.4, 26.5, 32.9, 47.4, 55.5, 1 13.2, 120.8, 122.7, 127.6, 1 34.1 , 1 55.9; MS (CI with NH3) m/e 206 (M+1 ); Anal. Calcd for CI 3H19NOxHBr: C, H, N; [α]D ²¹ = -58.4 (c=0.76 in MeOH).

S-(- 5-rr(Trifluoromethvnsulfonvlloxv1-2-(n- propvlamino etralin

Triflation of S-(-)-5-Hydroxy-2-(n-propylamino)tetralinxHBr (246 mg, 0.86 mmol) was performed according to the procedure given for the synthesis of R-(+)-8-[[(Trifluoromethyl)sulfonyl]oxy]-2-(n- propylamino)tetralin above, giving of an oil (248 mg) after extractive workup which was purified on a silica column, eluting with CH2Cl2/MeOH (1 :1 ). Conversion to the HCl salt and subsequent recrystallization from methanol/ether gave white crystals (1 90 mg, 59%): mp 232-233 °C (HCl); IR (KBr) 121 3 cm-1 ; I H NMR (CD30D) 1.00 (t, J = 7.5, 3H), 1.54-1.73 (m, 3H), 2.1 9-2.32 (m, I H), 2.65-2.86 (m, 4H), 2.97-32.26 (m, 3H), 7.1 2-7.28 (m, 3H); 1 3C NMR (CD30D) 1 1.0, 20.6, 23.1 , 25.7, 32.4, 47.8, 54.6, 1 19.7 (q, J = 319, CF3), 1 20.2, 128.7, 129.2, 130.4, 137.1 , 148.9; MS (CI with NH3) m/e 338 (M+1 ); Anal. Calcd for C14H1 8N03SF3xHCI: C, H, N; [α]D ²¹ = -62.4 (HCl) (c = 1.0 in MeOH).

5-Methoxy-3-(N-propylamino chromane: RN Ti l 9755-71 -41 and RN

Ti l 2904-79-71

SUBSTITUTE SHEET

5-Methoxy-3-aminochromanexHCI (RN [1 19755-62-3] and [1 10927- 03-2]; described in Thorberg et al., Acta Pharm. Suec, 24 (4), 1 69- 82, 1 987) (26 mg, 0.1 2 mmol) was propionylated with propionylchloride (1 6 μL, 1 .5 equiv.) in CH2 CI2 (2 mL) in the presence of Et3N (34 μL, 1.5 equiv.). The reaction was quenched after 2 h with MeOH (0.5 mL) and the mixture was worked up by adding CH2CI2 (20 mL) and washing the organic layer with water (1 0 mL). The organic layer was separated, dried (MgSθ4) and filtered and the solvent was evaporated under reduced pressure yielding an oil (28 mg, 98%).

This amide (32 mg, 0.14 mmol) was dissolved in dry THF (2 mL) and reduced with LiAIH4 (27 mg, 5 equiv.) under N2 (g) at room temperature. After 3 h the reaction was stopped by the addition of 30 μL H2O, 30 μL 10% NaOH and 90 μL H2O. The solid material was filtered and the solvent of the filtrate was evaporated under reduced pressure, yielding a colourless oil (28 mg, 93%). Conversion to the HCl salt gave white crystals (32 mg, 89%). GC/MS shows: M ⁺ (also base peak) at m/e = 221 . Other prominent peaks were found at: m/e = 1 92 (M-29; CH2CH3) and m/e = 163 (M- NHCH2CH2CH3).

5-Methoxy-3-(N-butylamino chromane

5-Methoxy-3-aminochromanexHCI (RN [1 19755-62-3] and [1 10927-

03-2]; described in Thorberg et al., Acta Pharm. Suec, 24 (4), 1 69-

82, 1 987) (100 mg, 0.47 mmol) was amidated with buturylchloride

(75 μL, 1 .5 equiv.) in CH2CI2 (6 mL) in the presence of Et3N (1 30

SUBSTITUTE SHEET

μL, 2.0 equiv.). The reaction was quenched after 3 h with MeOH (0.5 mL) and the mixture was worked up by adding CH2CI2 (20 mL) and washing the organic layer with water (1 0 mL). The organic layer was separated, dried (MgSθ4) and filtered and the solvent was evaporated under reduced pressure yielding a brownish solid (1 1 3 mg, 97%). GC/MS shows: M+ (also base peak) at m/e = 249. Another prominent peak (base peak) was found at: m/e = 1 62 (M-H- NHCH2CH2CH2CH3, i.e. elimination of the amide functionality).

This amide (1 04 mg, 0.42 mmol) was dissolved in dry THF (5 mL) and reduced with L.AIH4 (75 mg, 1.97 mmol) under N2(g) at room temperature. After 3 h the reaction was stopped by the addition of 75 μL H20, 75 μL 10% NaOH and 1 50 μL H2O. The solid material was filtered and the solvent of the filtrate was evaporated under reduced pressure, yielding a yellow oil (93 mg, 94%). GC/MS shows: M+ (also base peak) at m/e = 235. Other prominent peaks were found at: m/e = 1 92 (M-CH2CH2CH3) and m/e = 1 63 (M- NHCH2CH2CH2CH3).

5-Hvdroxy-3-(N-butvlamino ^, )chromane

5-Methoxy-3-(N-butylamino)chromanexHCI (105 mg, 0.39 mmol) was dissolved in HOAc (2 mL) and 48% HBr (1 mL) was added. The rection mixture was refluxed under N2(g) for 2 h and was then allowed to cool to room temperature. The reaction mixture was poured into saturated Na2C03 (50 mL) and the product was extracted with CH2CI2 (1 x50 mL and 2x25 mL). The combined organic layers were dried (Na2S04), filtered and the solvent was

SUBSTITUTE SHEET

evaporated under reduced pressure to give a brown oil (77 mg, 84%), which was chromatographed on Siθ2, using CH2θ2/MeOH (7 .1 ) as eluent. The fraction containing pure product were poole and the solvent was evaporated under reduced pressure to give a colourless oil (48 mg, 56%). GC/MS shows: M+ at m/e = 221. Other prominent peaks were found at: m/e = 178 (M-CH2CH2CH3) and m/e = 149 (M-NHCH2CH2CH2CH3).

5-Hvdroxv-3-(N-propvlamino chromane

5-Methoxy-3-(N-propylamino)chromanexHCI was hydrolyzed in HOAc and 48% HBr , as described for the preparation of 5-hydroxy- 3-(N-butylamino)chromane above. GC/MS shows: M ⁺ at m/e = 221. Other prominent peaks were found at: m/e = 192 (M-CH2CH3) and m/e = 163 (M-NHCH2CH2CH3).

SUBSTITUTE SHEET

The following compounds (sulphonates), all racemic, have been synthesized from the corresponding phenols under phase transfer catalyst conditions (QHSO4 , CH2CI2 and 1 0% NaOH in room temperature), using the appropriate sulphonylchlorides (purchased from Aldrich).

structure direct inlet (CD or GC/MS (El- m

TETRALINS

OS0 ₂ CH ₃

M+ at m/e=283, base peak at m/e=254 (M-CH2CH3) and other peaks at m/e=225 (M-NHCH2CH2CH3) and m/e=204 (M-CH3S02).

CI: M+1 at m/e=346

SUBSTITUTE SHEET

46

CI: M+1 at m/e=362

CI: M+1 at m/e=354

CI: M+1 at m/e=298.

SUBSTITUTE SHEET

CI: M+1 at m/e=352.

CI: M+1 at m/e=360.

CI: M+1 at m/e=374.

SUBSTITUTE SHEET

CI: M+1 at m/e=298.

OS0 ₂ CH _Ξ

CI: M+1 atm/e=284.

CI: M+1 atm/e=346.

SUBSTITUTE SHEET

I: M+1 at m/e=352.

M+ at m/e=273, base peak at m/e=176 (M-CH2C4H3S) and another peak at m/e=147 (M-NHCH2CH2C4H3S).

SUBSTITUTE SHEET

M+ at m/e=351 , M-1 at 350, base peak at m/e=254 (M-CH2C4H3S) and another peak at m/e=225 (M-NHCH2CH2C4H3S).

M+ at m/e=405, M-l at 404, base peak at m/e=308 (M-CH2C4H3S) and anotner peak at m/e=279 (M-NHCH2CH2C4H3S).

No M+ peak, base peak at m/e=316 (M-CH2C4H3S) and another peak at m/e=287 (M-NHCH2CH2C4H3S); CI gave M+1 =414. SUBSTITUTE SHEET

I: M+1 at m/e=420.

SUBSTITUTE SHEET

CHROMANES

OS0 ₂ CH ₃

CI: M+1 at m/e=286.

OSC\CF ₃

I: M+1 at m/e=340.

I: M+1 at m/e=348

SUBSTITUTE SHEET

CI: M+1 at m/e=362.

CI: M+1 at m/e=354.

OS0 ₂ CH ₃

M+ at m/e=299, base peak at m/e=256 (M-CH2CH2CH3) and another peak at m/e=220 (M-S02CH3).

SUBSTITUTE SHEET

OS0 ₂ CF ₃

M ⁺ at m/e=353, base peak at m/e=220 (M-S02CF3) and other peaks at m/e=310 (M-CH2CH2CH3) and m/e=281 (M-NHCH2CH2CH2CH3).

M+ at m/e=361 , base peak at m/e=220 (M-S02C6H6) and other peaks at m/e=31 8 (M-CH2CH2CH3) and m/e=289 (M- NHCH2CH2CH2CH3).

SUBSTITUTE SHEET

M+ at m/e=375, base peak at m/e=220 (M-S02C6H4CH3) and other peaks at m/e=332 (M-CH2CH2CH3 ) and m/e=303 (M- NHCH2CH2CH2CH3).

M+ at m/e=367, base peak at m/e=220 (M-S02C4H3S) and other peaks at m/e=324 (M-CH2CH2CH3) and m/e=295 (M- NHCH2CH2CH2CH3).

CI: M+1 at m/e=236.

CI: M+1 at m/e=208.

SUBSTITUTE SHEET

Pharmacology

Blochemsitry. Rats (1 50-300 g) pretreated with reserpine (5 mg/kg, 1 8 hours before) are given the test compounds. Gross behavioral observations (changes in motility, hindleg abduction, etc.) are made. Subsequent administration of NSD 101 5, decapitation, brain dissection (corpora striata, the limbic forebrain and the remaining hemispheral portions (mainly cortex) or rat brain), homogenization, centrifugation, ion-exchange chromatography and spectrofluorimetric measurements (all as described in detail by Wikstrόm, et al., J. Med. Chem., 21 , 864-867, 1 978 and references cited therein), or by HPLC/EC, gave the actual DOPAC and 5-HTP levels. Several doses (n=4-6) are tested for each compound and brain area. The dose of a compound giving 50% of the maximal reduction of the 5-HTP level in the rat brain part is then estimated.

The compounds of this invention are both behaviorally and biochemically active, producing the above mentioned effects, indicating either central DA or 5-HT receptor stimulation. The absence of significant decreases in the DOPAC levels in the hemispheral brain parts suggests that the compounds do not possess central NA receptor stimulating effects at the dosage under consideration.

SUBSTITUTE SHEET

Biochemical and behavioral effects of 5-, 6-, 7- and 8-OTf-PAT have been published in Sonesson et al. in J. Med. Chem., 1 993, 36, 3409-1 6. Biochemical and behavioral effects of cis-(+)-1 -Me-5- OTf-PAT has been published in Haadsma-Svensson et al. in Bioorg. & Med. Chem. Let., 1993, 36, 3409-16.

Behavioral anti-anxiety tests.

Behavioral approaches to studying the role of 5HT in anxiety involves animal models that are based on simple adaptions to aversive or threatening stimuli. One example is the 'elevated plus- maze test' (Pellow et al., 1 985). Normally, rats restrict their activity to the enclosed area of the plus-maze, avoiding the two open arms. This behavior can be suppressed by anxiolytic drugs and enhanced by anxiogenics. The plus-maze is relatively insensitive to drugs other than anxiolytics and anxiogenics and provides a simple test of anxiety in rats (Griebel et al., 1993). Prior exposure to an emotional stressor produces higher emotionality in the animals which is reflected by reduced exploration of the open maze arms in favor of enclosed maze arms (Heinrichs et al.,1992).

A second simple animal model of anxiety is the 'conditioned defensive burying test' (Treit et al., 1981 ). In this test, rats are exposed to an electrified shock-probe, and the duration of burying behavior is the major index of anxiety. Standard anti-anxiety agents suppress this burying response in a dose-related manner.

SUBSTITUTE SHEET

The combination of these tests is particularly important (Treit et al., 1 993). In the plus-maze test, passive avoidance of novel, elevated, open platforms is the major index of 'anxiety', while in the conditioned defensive burying test, the major index of 'anxiety' is the active burying of the shock-probe.

Animals

Male Wistar rats (n=5-9) weighing 300-500 g at the beginning of the experiments were used. They were housed individually in clear Plexiglass cages (25 x 25 x 30 cm) on a 12 h light-dark regime (light on between 08.00 and 20.00). All animals had free access to standard rat chow (Hope Farms) and tapwater. The experiments were carried out between 10.00 and 14.00.

R-(+)-8-[[(Trifluoromethyl)sulfonyl]oxy]-2-(n-propyl- amino)tetralin (Sonesson et al., 1993) was dissolved in saline which, alone, served for control injections. The compound was given intraperitoneally (i.p.) 30 min before the test session in a dose range of 1.0-3.0 mg/kg . Cited doses refer to the HCl salt, and neither of these doses did produce the 5HT-syndrome in the rats.

Conditioned defensive burying

The Shock-probe defensive burying test was performed in the home cage of the animals. The floor was covered with wood shavings

(height 2 cm). A removable teflon probe (6.5 cm long, 1 cm in diameter) was positioned 2 cm above the bedding. The probe was

SUBSTITUTE SHEET.

inserted through a small hole in the center of the longest wall of the cage. Two exposed wires (0.5 mm in diameter) were each wrapped (25 times) independently around the probe. Whenever the animal touched both wires simultaneously, with any part of its body, an electric current of 1 .5 mA was delivered to the animal. During the entire period the shock circuit was left on, i.e. "repeated shock probe procedure" was used (Treit and Fundytus, 1 988). Shock intensity was adjusted with a variable resistor in series with a 1000 8V shock source. On day 2 vehicle or drug was injected 30 min before introduction of the non-electrified probe in the home cages of the rats. Thus the procedure investigated the conditioned emotional consequence of former punishment rather than the direct effect of shock. All animals were observed for 10 min. Animals burying less than 25% of total time on day 1 were not tested for day 2.

Fear of footshock

The rats were trained in a passive avoidance apparatus in which the electric shock would be applied (Korte et al., 1990). Rats were exposed to a dark compartment where they were allowed to stay for 5 min. During the trial an inescapable scrambled footshock (0.6 mA, AC for 3 s) was given after the minutes 1 and 4. On the next day the animals are reexposed to the dark compartment in which no further footshock was given.

Elevated Plus-Maze

SUBSTITUTE SHEET

Directly after the 5 min period exposure to the shock compartment (above), the animals were placed in the elevated Plus-maze. The elevated Plus-maze (wood, painted black) consisted of two open, 50x1 0 cm, and two enclosed arms, 50x10x40 cm, with an open roof, arranged such that the two arms of each type were opposite to each other (Pellow et al., 1985), connected by an open central area (10 1 0 cm) . The maze was elevated to a height of 50 cm above the floor. Rats were individually placed in the centre of the maze facing an enclosed arm. The maze was cleaned after each rat.

Behavioral Measurements

During the exposure to the former shock compartment time spent immobile, i.e. when the animal is completely motionless, was measured. Time spent in the open arms relative to the time spent in the open plus the closed arms, is used as an index for the anxiolytic or anxiogenic effects (Pellow et al., 1985; Wahlestedt et al., 1 993). Furthermore, the activity score, i.e. number of arm entries, the closed arm entries and the open arm entries were measured. The observations were recorded by trained observers who were unaware the the treatment schedule.

The behaviour in the defensive burying paradigm was classified in five categories: (a) defensive burying - moving toward the probe and spraying or pushing bedding material toward the probe with rapid movements of the snout or forepaws as described earieir by

Pinel and Treit (1983).; (b) eating, chewing chow or faeces; (c) rearing, standing or sitting on hindlegs, mostly making sniffinq

SUBSTITUTE SHEET

movements, pointing the nose up into the air; (d) resting, the hindlimbs, the forelimbs, and the belly of the rat touched the floor and supported its body weight; (e) exploring, investigation of any part of the home cage.

Statistics

The data were analyzed with oneway ANOVA's. The ANOVA's were followed by the Dunnett's t-test in order the compare the vehicle group to each of the other groups.

SUBSTITUTE SHEET

Results

Conditioned defensive burying

The burying behavior after 3 mg/kg, but not after 1 mg/kg administration of R-(+)-8-[[(trifluoromethyl)sulfonyl]oxy]-2-(n- propyl-amino)tetralin (Sonesson et al., 1 993), is completedly abolished (F(2,1 8)=3.46, p=0.057) whereas the % rearing is also dose-dependently decreased. Interestingly, both the 1 mg/kg and the 3 mg/kg dose dramatically increased the food intake.

Elevated plus-maze

One-factor ANOVA revealed a significant effect of R-(+)-8- [[(trifluoromethyl)sulfonyl]oxy]-2-(n-propyl-amino)tetralin

(Sonesson et al., 1993) on the number of open arm entries at 3 mg/kg (F(2,21 )=1 5.27; p<0.01 ) and on the total number of entries (F(2,21 )=3.97; p<0.05). Furthermore, the number of open arm entries between the non-stressed and stressed animals is also significantly different (p<0.05, T-test). The % time L/L+D shows the same trend as the open arm and total entries whereas the number of enclosed arm entries is not significantly changed at the applied dose-range.

Without further elaboration, it is believed that one skilled in the art can, using the preceding description, practice the present invention to its fullest extent. The above mentioned examples describe how to prepare the various compounds and/or perform the

SUBSTITUTE SHEET

various processes of the invention and are to be construed as merely illustrative, and not limitations of the preceding disclosure in any way whatsoever. Those skilled in the art will promptly recognize appropriate variations from the procedures both as to reactants and as to reaction conditions and techniques.

References for the Pharmacology section

Beer, M., Kennett, G.A. and Curzon, G., 1990, A single dose of 8-OH- DPAT reduces raphe binding of [3 H]8-OH-DPAT and increase the effect of raphe stimulation of 5HT metabolism, European J. Pharmacol. 178, 179.

Blier, P. and de Montigny, C, 1987, Modifications of 5HT neuron properties by sustained administration of the 5HT-| A agonist gepirone: Electrophysiological studies in the rat brain, Synapse 1 , 470.

Dourish, C.T., Hutson, P.H. and Curzon, G., 1 986, Para- chlorophenylalanine prevents feeding induced by the serotonin agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT), Psychopharmacology 89, 467.

Griebel, 1 993

Heinrich, 1992

SUBSTITUTE SHEET

Higgins, G.A., Bradbury, A.J., Jones, B.J. and Oakley, N.R., 1 988, Behavioral and biochemical consequences following activation of 5HT-| -like and GABA receptors in the dorsal raphe nucleus of the rat, Neuropharmacology 96, 829.

Higgins, G.A., Jones, B.J. and Oakley, N6.R., 1992, Effect of 5HTI A receptor agonists in two models of anxiety after dorsal raphe injection, Psychopharmacology 106, 261.

Kennett, G.A., Marcou, M., Dourish, C.T. and Curzon, G., 1987, Single administration of 5HTI A agonists decreases 5HTI A presynaptic, but not postsynaptic receptor-mediated responses: relationship to antidepressant-like action, European J. Pharmacol. 138, 53.

Pellow, S., Chopin, P., File, S.E. and Briley, M., 1985, Validation of open:closed arm entries in elevated plus-maze as a measure of the anxiety in the rat, J. Neurosci. Method. 144, 149.

Pinel and Treit, 1983

Sonesson, C, Boije, M., Svensson, K., Ekman, A., Carlsson, A., Romero, A.G., Martin, I.J., Duncan, J.N., King, L.J. and Wikstrόm, H., 1 993, Orally active central dopamine and serotonin receptor ligands: 5-, 6-, 7-, and 8-[[(Trifluoromethyl)sulfonyl]oxy]-2-(di-n- propylamino)tetralins and the formation of active metabolites in vivo, J. Med. Chem. 36, 3409.

SUBSTITUTE SHEET

Sprouse, J.S. and Aghajanian, G.K., 1 987, Electrophysiological response of serotonergic dorsal raphe neurones to 5HTι A ^ana " 5HT-| B agonists, Synapse 1 , 9.

Treit, D., Pinel, J.P.J. and Fibiger, H.C., 1981 , Conditioned Defensive burying: a new paradigm for the study of anxiolytic agents, Pharmacol. Biochem. Behav. 1 5, 619.

Treit, D. and Fundytus, M., 1 988, A comparison of buspirone and chlordiazepoxide in the shock-probe/burying test for anxiolytics, Pharmacol. Biochem. Behav. 31 , 1071.

Treit, D., Robinson, A., Rotzinger, S. and Pesold C, 1993, Anxiolytic effects of serotonergic interventions in the shock-probe burying test and the elevated plus-maze test, Behav. Brain Res., 54, 23.

Vandermaelen, C.P., Matheson, G.K., Wilderman, R.C. and Patterson, L.A., 1 986, Inhibition of serotonergic dorsal raphe neurons by systemic and iontophoretic administration of buspirone, a non- benzodiazepine anxiolytic drug, European J. Pharmacol. 129, 123.

Wahlestedt, 1 993

SUBSTITUTE SHEET