Indian Patents. 234574:"ISOXAZOLINE COMPOUNDS"

Title of Invention	"ISOXAZOLINE COMPOUNDS"
Abstract	Isoxazoline compounds of general Formula (I) the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the N-oxide form thereof.

Full Text	The present invention relates to isoxazoline compounds. The invention concerns substituted isoxazolines derivatives having anti-depressant activity and/or anxiolytic activity and/or body weight control activity, processes for their preparation, pharmaceutical compositions comprising them and their use as a medicine, in particular for the treatment of depression, anxiety, stress-related disorders associated with depression and/or anxiety and disorders of body weight including anorexia nervosa and bulimia. The invention also relates to novel combination of substituted isoxazolines derivatives having anti-depressant activity and/or anxiolytic activity and/or body weight control activity with, anti depressants, anxiolytics and/or antipsychotics. Tetrahydronaphtalene and indane derivatives showing anti-depressant activity are known from EP-361 577 Bl. These compounds are typical monoamine reuptake blockers with additional (X2-adrenoceptor antagonist activity and they show antidepressant activity without being sedative. The problems associated with the compounds according to the state of the art is that the compounds cause considerable side-effects, such as nausea, excitation, an increased heart rate and a reduced sexual function. Furthermore, it requires a long time, in particular 3-4 weeks, before the response starts. The purpose of the present invention is to provide novel compounds derivatives having anti-depressant and/or anxiolytic and/or body weight control activity, in particular compounds that do not exhibit the aforementioned disadvantages. The present invention relates to novel isoxazoline derivatives according to the general (Formula Removed) the pharmaceuticalJy acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the N-oxide form thereof, wherein : X is CH2, N-R7, S or O ; R7 is selected from the group of hydrogen, alkyl, phenyl, phenylalkyl, alkylcarbonyl, alkyloxycarbonyl and mono- and dialkylaminocarbonyl, the phenyl and alkyl groups being optionally substituted with one or more halo atoms; R1 and R2 are each, independently from each other, selected from the group of hydrogen, hydroxy, cyano, halo, OSO2H, OS02CH3, phenyl, phenylalkyl, alkyloxy, alkyloxyalkyloxy, alkyloxyalkyloxyalkyloxy, tetrahydrofuranyloxy, alkylcarbonyloxy, alkylthio, alkyloxyalkylcarbonyloxy, pyridinylcarbonyloxy, alkylcarbonyloxyalkyloxy, alkyloxycarbonyloxy, alkenyloxy, alkenylcarbonyloxy and mono-and dialkylaminoalkyloxy, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; or R1 and R2 may be taken together to form a bivalent radical -R'-R2- selected from the group of -CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-O-, -CH2-O-CH2- and -0-CH2-CH2-O- ; a and b are asymmetric centers ; (CH2)m is a straight hydrocarbon chain of m carbon atoms, m being an integer ranging from 1 to 4 ; Pir is an optionally substituted radical according to any one of Formula (Ha)wherein : each R8 is independently from each other, selected from the group of hydrogen, hydroxy, amino, nitro, cyano, halo and alkyl; n is an integer ranging from 1 to 5 ; R9 is selected from the group of hydrogen, alkyl and formyl; and R3 represents an optionally substituted aromatic homocyclic or heterocyclic ring system together with an optionally substituted and partially or completely hydrogenated hydrocarbon chain of 1 to 6 atoms long with which said ring system is attached to the Pir radical and of which may contain one or more heteroatoms selected from the group of O, N and S. More in particular, the invention relates to compounds according to Formula (I) wherein R3 is a radical according to any one of Formula (Ilia), (Hlb) and (Hie) (Figure Removed) wherein : d is a single bond while Z is a bivalent radical selected from the group of -CHr, -C(=0)-, -CH(OH)-, -C(=N-OH)-, -CH(alkyl)-, -O-, -S-, -S(=O), -NH- and -SH-; or d is a double bond while Z is a trivalent radical of formula =CH- or =C(alkyl)-; A is a 5- or 6-membered aromatic homocyciic or heterocyclic ring, selected from the group of phenyl, pyranyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl, oxadiazolyl and isoxazolyl; p is an integer ranging from 0 to 4 ; q is an integer ranging from 0 to 7 ; R4 is selected from the group of hydrogen, alkyl, phenyl, biphenyl, naphthyl, halo and cyano, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; R5 is equal to R4 ; or R4 and R5 may be taken together to form a bivalent radical -R4-R5- selected from the group of -CH2-, =CH-, -CH2-CH2-, -CH=CH-, -O-, -NH-, =N-, -S-, -CH2N(-alkyl)-, -CH=N-, -CH2O- and -OCH2-; each R6 is independently from each other, selected from the group of hydrogen, hydroxy, amino, nitro, cyano, halo, carboxyl, alkyl, phenyl, alkyloxy, phenyloxy, alkylcarbonyloxy, alkyloxycarbonyl, alkylthio, mono- and dialkylamino, alkylcarbonylamino, mono- and dialkylaminocarbonyl, mono- and dialkylaminocarbonyloxy, mono- and dialkylaminoalkyloxy, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; or two vicinal radicals R6 may be taken together to form a bivalent radical -R6-R6- selected from the group of-CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-C(=O)-, -0-CH2-O-, -CH2-0-CH2-, -0-CH2-CH2-O-, -CH=CH-CH=CH-, -CH=CH-CH=N-, -CH=CH-N=CH-, -CH=N-CH=CH-, -N=CH-CH=CH-, -CH2-CH2-CH2-, -CH2-CH2-C(=O)- and -CH2-CH2-CH2-CH2-; and R10 is selected from the group of hydrogen, alkyl, phenylalkyl and phenyl. Preferably, the invention relates to those compounds wherein X=O or NH ; R1 and R2 are both alkyloxy ; m = 1; Pir is a radical according to Formula (Ila) wherein R8 is hydrogen and n = 4 ; R3 is a radical according to Formula (mb) wherein Z is =CH-, d is a double bond, A is a phenyl ring, R4 is an alkyl and R1 is hydrogen. More preferably, the invention relates to compounds where X=O, R1 and R2 are both methoxy ; m = 1; Pir is a radical according to Formula (Ha) wherein R8 is hydrogen and n = 4 ; R3 is a radical according to Formula (ITlb) wherein Z is =CH-, d is a double bond, A is a phenyl ring, R4 is methyl and R10 is hydrogen In the framework of this application, alkyl defines straight or branched saturated hydrocarbon radicals having from 1 to 6 carbon atoms, for example methyl, ethyl, propyl, butyl, 1-methylpropyl, 1,1-dimethylethyl, pentyl, hexyl; or alkyl defines cyclic saturated hydrocarbon radicals having from 3 to 6 carbon atoms, for example cyclopropyl, methylcyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Halo is generic to fluoro, chloro, bromo and iodo. Alkyl radicals being optionally substituted with one or more halo atoms are for example polyhaloalkyl radicals, for example difluoromethyl and trifluoromethyl. The pharmaceutically acceptable salts are defined to comprise the therapeutically active non-toxic acid addition salts forms that the compounds according to Formula (I) are able to form. Said salts can be obtained by treating the base form of the compounds according to Formula (I) with appropriate acids, for example inorganic acids, for example hydrohalic acid, in particular hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid and phosphoric acid ; organic acids, for example acetic acid, hydroxyacetic acid, propanoic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicyclic acid, p-aminosalicylic acid and pamoic acid. The compounds according to Formula (I) containing acidic protons may also be converted into their therapeutically active non-toxic metal or amine addition salts forms by treatment with appropriate organic and inorganic bases. Appropriate base salts forms comprise, for example, the ammonium salts, the alkaline and earth alkaline metal salts, in particular lithium, sodium, potassium, magnesium and calcium salts, salts with organic bases, e.g. the benzathine, N-methyl-D-glucamine, hybramine salts, and salts with amino acids, for example arginine and lysine. Conversely, said salts forms can be converted into the free forms by treatment with an appropriate base or acid. The term addition salt as used in the framework of this application also comprises the solvates that the compounds according to Formula (I) as well as the salts thereof, are able to form. Such solvates are, for example, hydrates and alcoholates. The N-oxide forms of the compounds according to Formula (I) are meant to comprise those compounds of Formula (I) wherein one or several nitrogen atoms are oxidized to the so-called N-oxide, particularly those N-oxides wherein one or more nitrogens of the piperazinyl radical are N-oxidized. The term "stereochemically isomeric forms" as used hereinbefore defines all the possible isomeric forms that the compounds of Formula (I) may possess. Unless otherwise mentioned or indicated, the chemical designation of compounds denotes the mixture of all possible stereochemically isomeric forms, said mixtures containing all diastereomers and enantiomers of the basic molecular structure. More in particular, stereogenic centers may have the R- or S-configuration; substituents on bivalent cyclic (partially) saturated radicals may have either the cis- or trans-configuration. Compounds encompassing double bonds can have an E or Z-stereochemistry at said double bond. Stereochemically isomeric forms of the compounds of Formula (I) are obviously intended to be embraced within the scope of this invention. Following CAS nomenclature conventions, when two stereogenic centers of known absolute configuration are present in a molecule, an R or S descriptor is assigned (based on Cahn-Ingold-Prelog sequence rule) to the lowest-numbered chiral center, the reference center. The configuration of the second stereogenic center is indicated using relative descriptors [R,R J or [R,S], where R* is always specified as the reference center and [RS] indicates centers with the same chirality and [R,S] indicates centers of unlike chirality. For example, if the lowest-numbered chiral center in the molecule has an 5 configuration and the second center is R, the stereo descriptor would be specified as S-[R,S]. If "a" and "(3" are used : the position of the highest priority substituent on the asymmetric carbon atom in the ring system having the lowest ring number, is arbitrarily always in the "a" position of the mean plane determined by the ring system. The position of the highest priority substituent on the other asymmetric carbon atom in the ring system (hydrogen atom in compounds of Formula® ) relative to the position of the highest priority substituent on the reference atom is denominated "a", if it is on the same side of the mean plane determined by the ring system, or "B", if it is on the other side of the mean plane determined by the ring system. Compounds of Formula (I) and some of the intermediates have at least two stereogenic centers in their structure, respectively denoted a and b in Formula (I). Due to the synthetic pathway followed for the synthesis of the tricyclic system, the configuration of those two asymmetric centers a and b is predetermined, so that the relative configuration of center a is S* and of center b is R. The invention also comprises derivative compounds (usually called "pro-drugs") of the pharmacologically-active compounds according to the invention, which are degraded in vivo to yield the compounds according to the invention. Pro-drugs are usually (but not always) of lower potency at the target receptor than the compounds to which they are degraded. Pro-drugs are particularly useful when the desired compound has chemical or physical properties that make its administration difficult or inefficient. For example, the desired compound may be only poorly soluble, it may be poorly transported across the mucosal epithelium, or it may have an undesirably short plasma half-life. Further discussion on pro-drugs may be found in Stella, V. J. et al., "Prodrugs", Drug Delivery Systems, 1985, pp. 112-176, and Drugs, 1985,29, pp. 455-473. Pro-drugs forms of the pharmacologically-active compounds according to the invention will generally be compounds according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the N-oxide form thereof, having an acid group which is esterified or amidated. Included in such esterified acid groups are groups of the formula -COOR, where Rx is a Chalky], phenyl, benzyl or one of the following groups : Amidated groups include groups of the formula - CONRyRz, wherein Ry is H, l, phenyl or benzyl and Rz is -OH, H, Chalky!, phenyl or benzyl. Compounds according to the invention having an amino group may be derivatised with a ketone or an aldehyde such as formaldehyde to form a Mannich base. This base will hydrolyze with first order kinetics in aqueous solution. The compounds of Formula (I) as prepared in the processes described below may be synthesized in the form of racemic mixtures of enantiomers that can be separated from one another following art-known resolution procedures. The racemic compounds of Formula (I) may be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently separated, for example, by selective or fractional crystallization and the enantiomers are liberated therefrom by alkali. An alternative manner of separating the enantiomeric forms of the compounds of Formula (I) involves liquid chromatography using a chiral stationary phase. Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically. Preferably if a specific stereoisomer is desired, said compound would be synthesized by stereospecific methods of preparation. These methods will advantageously employ enantiomerically pure starting materials. The compounds according to the invention have surprisingly been shown to have selective serotonine (5-HT) reuptake inhibitor activity in combination with additional cc2-adrenoceptor antagonist activity and show a strong anti-depressant and/or anxiolytic activity and/or a body weight control activity without being sedative. Also, in view of their selective serotonine (5-HT) reuptake inhibitor as well as a2-adrenoceptor antagonist activity, compounds according to the invention are also suitable for treatment and/or prophylaxis in diseases where either one of the activities alone or the combination of said activities may be of therapeutic use. In particular, the compounds according to the invention may be suitable for treatment and/or prophylaxis in the following diseases : • Central nervous system disorders, including : • Mood disorders, including particularly major depressive disorder, depression with or without psychotic features, catatonic features, melancholic features, atypical features of postpartum onset and, in the case of recurrent episodes, with or without seasonal pattern, dysthymic disorder, bipolar I disorder, bipolar n disorder, cyclothymic disorder, recurrent brief depressive disorder, mixed affective disorder, bipolar disorder not otherwise specified, mood disorder due to a general medical condition, substance-induced mood disorder, mood disorder not otherwise specified, seasonal affective disorder and premenstrual dysphoric disorders. • Anxiety disorders, including panic attack, agoraphobia, panic disorder without agoraphobia, agoraphobia without history of panic disorder, specific phobia, social phobia, obsessive-compulsive disorder, posttraumatic stress disorder, acute stress disorder, generalized anxiety disorder, anxiety disorder due to a general medical condition, substance-induced anxiety disorder and anxiety disorder not otherwise specified. Stress-related disorders associated with depression and/or anxiety, including acute stress reaction, adjustment disorders (brief depressive reaction, prolonged depressive reaction, mixed anxiety and depressive reaction, adjustment disorder with predominant disturbance of other emotions, adjustment disorder with predominant disturbance of conduct, adjustment disorder with mixed disturbance of emotions and conduct, adjustment disorders with other specified predominant symptoms) and other reactions to severe stress. Dementia, amnesic disorders and cognitive disorders not otherwise specified, especially dementia caused by degenerative disorders, lesions, trauma, infections, vascular disorders, toxins, anoxia, vitamin deficiency or endocrinic disorders, or amnesic disorders caused by alcohol or other causes of thiamin deficiency, bilateral temporal lobe damage due to Herpes simplex encephalitis and other limbic encephalitis, neuronal loss secondary to anoxia/ hypoglycemia severe convulsions and surgery, degenerative disorders, vascular disorders or pathology around ventricle HI. Cognitive disorders due to cognitive impairment resulting from other medical conditions. Personality disorders, including paranoid personality disorder, schizoid personality disorder, schizotypical personality disorder, antisocial personality disorder, borderline personality disorder, histrionic personality disorder, narcissistic personality disorder, avoidant personality disorder, dependent personality disorder, obsessive-compulsive personality disorder and personality disorder not otherwise specified. Schizoaffective disorders resulting from various causes, including schizoaffective disorders of the manic type, of the depressive type, of mixed type, paranoid, disorganized, catatonic, undifferentiated and residual schizophrenia, schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, shared psychotic disorder, substance-induced psychotic disorder and psychotic disorder not otherwise specified. Akinesia, akinetic-rigid syndromes, dyskinesia and medication-induced parkinsonism, Gilles de la Tourette syndrome and its symptoms, tremor, chorea, myoclonus, tics and dystonia. Attention-deficit / hyperactivity disorder (ADHD). Parkinson's disease, drug-induced Parkinsonism, post-encephalitic Parkinsonism, progressive supranuclear palsy, multiple system atrophy, corticobasal degeneration, parkinsonism-ALS dementia complex and basal ganglia calcification. Dementia of the Alzheimer's type, with early or late onset, with depressed mood. • Behavioral disturbances and conduct disorders in dementia and the mentally retarded, including restlessness and agitation. • Extra-pyramidal movement disorders. • Down's syndrome. • Akathisia. • Eating Disorders, including anorexia nervosa, atypical anorexia nervosa, bulimia nervosa, atypical bulimia nervosa, overeating associated with other psychological disturbances, vomiting associated with other psychological disturbances and non-specified eating disorders. • AIDS-associated dementia. Chronic pain conditions, including neuropathic pain, inflammatory pain, cancer pain and post-operative pain following surgery, including dental surgery. These indications might also include acute pain, skeletal muscle pain, low back pain, upper extremity pain, fibromyalgia and myofascial pain syndromes, orofascial pain, abdominal pain, phantom pain; tic douloureux and atypical face pain, nerve root damage and arachnoiditis, geriatric pain, central pain and inflammatory pain. Neurodegenerative diseases, including Alzheimer's disease, Huntington's chorea, Creutzfeld-Jacob disease, Pick's disease, demyelinating disorders, such as multiple sclerosis and ALS, other neuropathies and neuralgia, multiple sclerosis, amyotropical lateral sclerosis, stroke and head trauma. Addiction disorders, including: • Substance dependence or abuse with or without physiological dependence, particularly where the substance is alcohol, amphetamines, amphetamine-like substances, caffeine, cannabis, cocaine, hallucinogens, inhalants, nicotine, opioids, phencyclidine, phencyclidine-like compounds, sedative-hypnotics, benzodiazepines and/or other substances, particularly useful for treating withdrawal from the above substances and alcohol withdrawal delirium. • Mood disorders induced particularly by alcohol, amphetamines, caffeine, cannabis, cocaine, hallucinogens, inhalants, nicotine, opioids, phencyclidine, sedatives, hypnotics, anxiolitics and other substances. • Anxiety disorders induced particularly by alcohol, amphetamines, caffeine, cannabis, cocaine, hallucinogens, inhalants, nicotine, opioids, phencyclidine, sedatives, hypnotics, anxiolitics and other substances and adjustment disorders with anxiety. • Smoking cessation. • Body weight control, including obesity. • Sleep disorders and disturbances, including : • Dyssomnias and/or parasomnias as primary sleep disorders, sleep disorders related to another mental disorder, sleep disorder due to a general medical condition and substance-induced sleep disorder. • Circadian rhythms disorders. • Improving the quality of sleep. • Sexual dysfunction, including sexual desire disorders, sexual arousal disorders, orgasmic disorders, sexual pain disorders, sexual dysfunction due to a general medical condition, substance-induced sexual dysfunction and sexual dysfunction not otherwise specified. The present invention thus also relates to compounds of Formula (I) as defined hereinabove, the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof, as well as the prodrugs thereof for use as a medicine. Further, the present invention also relates to the use of a compound of Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form thereof as well as the pro-drugs thereof for the manufacture of a medicament for treating depression, anxiety and body weight disorders or more generally any one of the diseases mentioned above. The compounds according to the invention may also be suitable as add-on treatment and/or prophylaxis in the above listed diseases in combination with antidepressants, anxiolytics and/or antipsychotics which are currently available or in development or which will become available in the future, to improve efficacy and/or onset of action. This is evaluated in rodent models in which antidepressants, anxiolytics and/or antipsychotics are shown to be active. For example, compounds are evaluated in combination with antidepressants, anxiolytics and/or antipsychotics for attenuation of stress-induced hyperthermia. The invention therefore also relates to a pharmaceutical composition comprising the compounds according to the invention and one or more other compounds selected from the group of antidepressants, anxiolytics and antipsychotics as well as to the use of such a composition for the manufacture of a medicament to improve efficacy and/or onset of action in the treatment of depression and/or anxiety. In vitro receptor and neurotransmitter transporter binding and signal-transduction studies can be used to evaluate the a2-adrenoceptor antagonism activity and serotonine (5-HT) reuptake inhibitor activity of the present compounds. As indices for central penetration and potency to block the a2-adrenoceptors and serotonin transporters, respectively, ex vivo a2-adrenoceptor and serotonin transporter occupancy can be used. As indices of (X2-adrenoceptor antagonism in vivo, the reversal of the loss of righting reflex, observed in rats after subcutaneous injection or oral dosage of the compound before intravenous medetomidine administration in rats can be used (medetomidinetest). As indices of serotonine (5-HT) reuptake inhibition activity, the inhibition of head-twitches and excitation in rats, observed after subcutaneous injection or oral dosage of the compound before subcutaneous p-chloroamphetamine administration in rats can be used (pCA-test). The invention also relates to a composition comprising a pharmaceutically acceptable carrier and, as active ingredient, a therapeutically effective amount of a compound according to the invention. The compounds of the invention or any subgroup thereof may be formulated into various pharmaceutical forms for administration purposes. As appropriate compositions there may be cited all compositions usually employed for systemically administering drugs. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound, optionally in addition salt form, as the active ingredient is combined in intimate admixture with a pharmaceutically acceptable carrier, which carrier may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are desirable in unitary dosage form suitable, in particular, for administration orally, rectally, percutaneously, by parenteral injection or by inhalation. For example, in preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid preparations such as suspensions, syrups, elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrating agents and the like in the case of powders, pills, capsules and tablets. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit forms in which case solid pharmaceutical carriers are obviously employed. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, though other ingredients, for example, to aid solubility, may be included. Injectable solutions, for example, may be prepared in which the carrier comprises saline solution, glucose solution or a mixture of saline and glucose solution. Injectable suspensions may also be prepared in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations that are intended to be converted, shortly before use, to liquid form preparations. In the compositions suitable for percutaneous administration, the carrier optionally comprises a penetration enhancing agent and/or a suitable wetting agent, optionally combined with suitable additives of any nature in minor proportions, which additives do not introduce a significant deleterious effect on the skin. Said additives may facilitate the administration to the skin and/or may be helpful for preparing the desired compositions. These compositions may be administered in various ways, e.g., as a transdermal patch, as a spot-on, as an ointment. It is especially advantageous to formulate the aforementioned pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form as used herein refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, suppositories, injectable solutions or suspensions and the like, and segregated multiples thereof. The compounds according to the invention can generally be prepared by a succession of steps, each of which is known to the skilled person. In particular, the compounds according to Formula (I) with a Pir-radical according to Formula (Ila) can be prepared by a nucleophilic substitution reaction with a substituted piperazine according to Formula (V) on an intermediate of Formula (IV). These reactions may be carried out in a reaction inert solvent such as dioxane, methylisobutylketone or N,N'-dimethylformamide, in the presence of a suitable base such as potassium carbonate, sodium carbonate or triethylamine, or even without a base, using in this latter case excess of reagent of Formula (V). Convenient reaction temperatures range between 100°C and 150°C. (Figure Removed) In compound (IV), L represents any suitable reactive leaving group, in particular halo, such as chloro, bromo or iodo or sulfonyloxy, such as methylsulphonyloxy or 4-methylbenzenesulfonyloxy. The compounds according to Formula (I) with a Pir-radical according to Formula (Ha) can also be prepared by a 2-step reaction scheme in which an intermediate of Formula (IV) is first reacted with a substituted piperazine according to Formula (VII) after which the R3-radical is introduced into the molecule. Reaction conditions are similar to those described above for compounds of Formula (VI). (Figure Removed) In compound (IV), L represents any suitable reactive leaving group, in particular halo, such as chloro, bromo or iodo or sulfonyloxy, such as methylsulphonyloxy or 4-methylbenzenesulfonyloxy. In intermediate compound (VII), one of the nitrogen function may also be protected, e.g. by a tert-butyloxycarbonyl-group. (step 2) (Figure Removed) In compound (IX), L represents any suitable reactive leaving group, in particular halo, such as chloro, bromo or iodo or sulfonyloxy, such as methylsulphonyloxy or 4-methylbenzenesulfonyloxy. Also R3"CHO may be used as compound (IX). The compounds according to Formula (I) with a Pir-radical according to Formula (Ha) can also be prepared by a 2-step reaction scheme in which an intermediate of Formula (VET) is reacted with an acid according to Formula (X), followed by a subsequent reduction of the carbonyl-function of intermediate (XI). Reactions of step 1 may be carried out in a reaction inert solvent, such as chloroform, dichloromethane, tetrahydrofuran, dimethylformamide or a mixture thereof, using any of methods known to a person skilled in the art using condensation reagents such as l,l'-carbonyldiimidazole, N,N'-dicyclohexylcarbodiimide or by previous transformation of carboxylic acid of Formula (X) into its corresponding acid chloride. Reactions shown in step 2 can be performed using a suitable reducing agent, such as lithium-aluminum hydride or aluminum hydride, in a suitable solvent, for example tetrahydrofuran. Generally, these reactions are run at a temperature ranging between -20°C and room temperature. (Figure Removed) In intermediate compounds (XI) and (XII), the A-group represents an optionally substituted aromatic homocyclic or heterocyclic ring system including a partially or completely hydrogenated hydrocarbon chain of maximum 5 atoms long of which one or more carbon atoms may be replaced by one or more atoms selected from the group of oxygen, nitrogen and sulphur, with which the ring system is attached to the Pir radical that has been defined above. The substituents R1 and R2 may be changed or interconverted into each other by methods well known in the art, such as demethylation, acylation, esterification, amination and amidation. The starting materials and some of the intermediates are compounds that are either commercially available or may be prepared according to conventional reaction procedures generally known in the art. For example, intermediates of Formula (IV) in which X=O may be prepared according to the following reaction scheme (Scheme 1): Scheme 1 In intermediate compound (XIV), L represents any suitable reactive leaving group, in particular halo, such as chloro, bromo or iodo or sulfonyloxy, such as methylsulphonyloxy or4-methylbenzenesulfonyloxy. Furthermore, Alk in intermediate compound (XIV) represents any C1-6alkyl-group, in particular an ethyl-group and m is defined as in Formula (I). Intermediates according to Formula (IV) in which X=NH may also be prepared in an equivalent manner according to above step 1, provided that the intermediate compound (Xin) is replaced by its amine-analog (XVI), preferably with the amine group protected with e.g. a COCFa- group. The alkylation step may be carried out in a reaction inert solvent, for example, tetrahydrofuran or dimethylformamide, in the presence of a strong base, such as sodium or potassium hydride, and the addition of a crown-ether, such as 18-crown-6 or 15-crown-5. Convenient reaction temperatures range between room temperature and 60°C. Intermediates of Formula (XVII) are converted to oximes of Formula (XVIQ) using artknown techniques, such as using hydroxylamine hydrochloride in the presence of NaHCO3 or pyridine in a reaction inert solvent, for example ethanol. Intermediate (XVIII) is oxidized to its nitril oxide and undergoes in situ an intramolecular cycloaddition, yielding compound of Formula (XIX). This oxidation can be carried out using a sodium hypochlorite solution in the presence of triethylamine in an inert solvent such as dichloromethane at room temperature. Oxidation can also be performed using Chloramine-T (N-chloro-4-methyl-benzenesulfonamide, sodium salt), stirring and heating in a solvent such as refluxing ethanol. At this stage the two stereocenters a and b of Formula (I) are formed. (Figure Removed) Preparation of a compound of Formula (XX) can be achieved using procedures known in the art, for instance by reduction of the carbonyl compound of Formula (XIX) in the presence of a suitable reducing agent, for example, sodiumborohydride in a suitable solvent, such as water, an alcohol, tetrahydrofuran or a mixture thereof, generally at room temperature. (Figure Removed) Intermediate of Formula (IV) can be prepared from intermediate of Formula (XX) using standard techniques. Thus, reaction with methanesulfonyl chloride or 4-methyIbenzenesulfonyl chloride in the presence of a base, such as triethylamine, in a reaction inert solvent, for example dichloromethane, at reaction temperatures ranging between 0°C and room temperature, yields the corresponding sulfonyloxy derivative intermediate (IV). The corresponding halo-derivative can also be prepared, e.g. treating intermediate of Formula (XX) with triphenylphosphine, in the presence of tetrachloromethane, in a reaction inert solvent, such as tetrahydrofuran, stirring and refluxing the mixture. (Figure Removed) It is evident that in the foregoing and in the following reactions, the reaction products may be isolated from the reaction medium and, if necessary, further purified according to methodologies generally known in the art such as extraction, crystallization and chromatography. It is further evident that reaction products that exist in more than one enantiomeric form, may be isolated from their mixture by known techniques, in particular preparative chromatography, such as preparative HPLC. Typically, intermediate compounds (IV) and final compounds according to Formula (I) may be separated into their enantiomeric forms. Compounds according to the invention in which X=CHz may be prepared according to the following reaction scheme (Scheme 2) in which an intermediary compound according to Formula (V) is first N-alkylated with a dihaloderivative of Formula (XX) using standard techniques, in the presence or absence of a base and in an inert reaction solvent, such as chloroform, dichloromethane or 1,2-dichloroethane, and at reaction temperatures ranging between room temperature and SOT, yielding an intermediate of Formula (XXI). An aldehyde of Formula (XXEf) was reacted with rerr-butylamine in an aprotic solvent such as toluene, stirring and heating at reflux temperature with removal of water using a standard device, such as a Dean-Stark water separator, yielding an imine of Formula (XXTV). C-alkylation of intermediary compound of Formula (XXIV) with intermediate of Formula (XXI) can be achieved in the presence of an alkyl-lithium derivative, such as n-butyllithium, under an inert atmosphere and in a dry inert solvent, such as tetrahydrofuran, at low temperatures ranging between -78°C and 0°C, yielding an intermediate of Formula (XXV). The intermediate compound of Formula (XXVI) may be prepared by reaction of compound of Formula (XXV) with hydroxylamine, in the presence of a base such as sodium bicarbonate, in a solvent such as a lower alkylalcohol like ethanol, generally at room temperature. Finally, the oxidation of the oxime derivative of Formula (XXVI) to its nitril oxide and subsequent in situ cycloaddition to give compound of Formula (XXVII), may be achieved by similar standard techniques such as those described above for intermediate of Formula (XVIII) to give compounds of Formula (XIX). (Figure Removed) (step 5) It is evident that the reaction steps disclosed above may be adapted to the specific reaction products. The reaction steps disclosed may be performed in any way known to the skilled person, including in solution or as solid phase reactions, the latter during which the reaction products are bound to a resin material and are - in a final cleavage step - released from the resin material. Examples of such embodiments and adaptations have been disclosed by way of the Examples further in this application. The compound 3,3a,4,5-tetrahydronaphto[l,2-c]isoxazole-3-acetic acid (Formula (IV) wherein each of R1 and R2 are H, m=0, X=CH2 and L=COOH) and has been disclosed in Synthetic Communications. 27(16), 2733-2742 (1997) as an intermediate for the syntheses of anti-inflammatory, analgesic and antipyretic compounds and is excluded from patent protection. The following examples illustrate the present invention without being limited thereto. Experimental part The carbon ring numbering system for the compounds according to Formula (I) used in this application is as follows : Pir ]— R3 Of some compounds the absolute stereochemical configuration of the stereogenic carbon atom(s) therein was not experimentally determined. In those cases the stereochemically isomeric form which was first isolated is designated as "A" and the second as "B", without further reference to the actual stereochemical configuration. However, said "A" and "B" isomeric forms can be unambiguously characterized by a person skilled in the art, using art-known methods such as, for example, X-ray diffraction. The stereogenic centers a and b in Formula (I) have respectively the ring numbers 3a and 3. Hereinafter, "DMF" is defined as A-dimethylformarnide, "DIPE" is defined as diisopropyl ether, and "THF" is defined as tetrahydrofurane. A. Preparation of the intermediate compounds Example Al.a Preparation of intermediate 1 A solution of 4-bromo-2-butenoic acid methyl ester (0.1647 mol) in DMF (50 ml) was added dropwise to a mixture of 2-hydroxy-4,5-dimethoxy-benzaldehyde (0.0823 mol) and K2CO3 (0.1647 mol) in DMF (200 ml). The reaction mixture was stirred for 2 hours at room temperature, filtered and the filtrate was evaporated to dryness. The residue was washed in a 10% aqueous NaOH solution, then extracted with QHkCl2. The separated organic layer was dried (Na2SC>4), filtered, and the solvent was evaporated. The residue was washed with diethyl ether, then dried. Yielding: 20 g of intermediate 1 (87%). Example Al.b Preparation of intermediate 2 Hydroxylamine (0.045 mol) was added to a solution of intermediate 1 (0.041 mol) in ethano] (150 ml). Pyridine (57 ml) was added. The reaction mixture was stirred for 2 hours at room temperature, then poured out into water and acidified with concentrated HC1. This mixture was extracted with CH2Ch. The separated organic layer was dried (NaaSCU), filtered, and the solvent was evaporated. Yielding: 11.7 g (96%, crude yield). A sample (2 g) was purified by high-performance liquid chromatography over silica gel (eluent: CHaC2/CHsOH 95/5). The pure fractions were collected and the solvent was evaporated. The residue was washed with diethyl ether, then dried. Yielding: 0.9 g intermediate 2 . Example Al.c (Figure Removed) Preparation of intermediate 3 NaOCl, 5% (130 ml) was added dropwise to a mixture of intermediate 2 (0.037 mol) and EtsN (1 ml) in CH2Cla (220 ml). The reaction mixture was stirred for 4 hours at room temperature, then washed with water, dried (NaaSC^), filtered, and the filtrate was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/2-propanone 100/0 and 95/5). The desired fractions were collected and the solvent was evaporated. Yielding: 5.8 g (54%, used in next traction step without further purification). A sample (2 g) was recrystallised from EtOAc. The precipitate was filtered off and dried. Yielding: 1.7 g of intermediate 3. Example Al.d Preparation of intermediate 4 NaBtLt (0.043 mol) was added portionwise to a solution of intermediate 3 (0.017 mol) in THF (50 ml) and H2O (5 ml), stirred and cooled on an ice-bath. The resulting reaction mixture was stirred for 2 hours at room temperature. 2-Propanone was added while stirring for 30 min. The reaction mixture was washed with water and extracted with CHzCh. The separated organic layer was washed with brine, dried filtered and the solvent evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2C12/CH3OH 95/5) and by high-performance liquid chromatography over silica gel (eluent: CH2C12/CH3OH 98/2). The pure fractions were collected and the solvent was evaporated. A sample (1.8 g) was treated with diethyl ether, then dried. Yielding: 1.2 g of intermediate 4 (59%). Example Al.e Preparation of intermediate 5 Et3N (0.016 mol) was added to a solution of intermediate 4 (prepared according to A3) (0.0109 mol) in CH2C12 (60 ml). The mixture was cooled in an ice-bath. Methanesulfonyl chloride (0.012 mol) was added and the resulting reaction mixture was stirred for 30 min. Then, the mixture was washed with water, dried (Na2SCO4), filtered and the solvent was evaporated. Yielding: 3.5 g of intermediate 5 (82%, used in next reaction step without further purification). Example Al.f Preparation o f intermediate 6 I T T Reaction under N2 atmosphere. BBr3 (0.04368 mol) was added dropwise to a stirred solution of intermediate 5 (prepared according to Al.e) (0.00873 mol) in CH2C12 (100 ml), cooled to -78 °C. The reaction mixture was allowed to warm to -40 °C and stirring was continued for 2 hours at -40 °C. Then, the mixture was poured out into ice-water and extracted with CH2C12. The separated organic layer was dried (MgSO4), filtered and the solvent evaporated. The residue was purified by flash column chromatography over silica gel (eluent: CH2C12/CH3OH 97/3), then by HPLC (eluent: CH2C12/CH3OH 99.5/0.5 to 90/10). Two product fraction groups were collected and their solvent was evaporated. Yield: 0.750 g of intermediate 6 (26%). Example Al.g Preparation of intermediate 7 A mixture of intermediate 5 (prepared according Aid) (0.0422 mol) and piperazine (0.1267 mol) in 1,4-dioxane (15 ml) was stirred for 2 hours at 100 °C. The solvent was evaporated. The residue was washed with water and extracted with CH2Cb- The separated organic layer was dried (NaaSO/t), filtered and the solvent was evaporated. Yielding: 13 g of intermediate 7 (NMR: 85%). Example Al.h Preparation of intermediate 8 Intermediate 5 (prepared according to Al.e) (200 g, 0.58 mol) was separated into its enantiomers by chiral column chromatography over column LCI 10-2 with stationary phase CfflRALPAK-AD (2000 g, packing pressure: 45 bar, detector range: 2.56, wavelength: 240nm, temperature: 30 °C; injection solution: 200 g in 8.4 L CH3CN; then, 19.6 L methanol (+ 2% ethanol) was added, then filtered; injection-volume: 700 ml; eluent: CH3OH/CH3CN 70/30 v/v). Two product fraction groups were collected and their solvent was evaporated. Yield: 95 g of intermediate 8 . Example Al.i Preparation of intermediate 9 A mixture of intermediate 8 (prepared according Al.h) (0.0728 mol) and l-(tertbutyloxycarbonyl) piperazine (0.087 mol) in dioxane (500ml) was stirred and refluxed for 48 hours. The solvent was evaporated and CH2Cla was added. HaO and NaOH (50%) were added also and the mixture was extracted with CH2C12. The separated organic layer was dried (MgSCU) and the solvent was evaporated in vacuum. Yield intermediate 9 Example Al.i Preparation of intermediate 10 A mixture of intermediate 9 (0.00318 mol) and 2,2,2-trifluoroacetic acid (189ml) in CH2Ck (500ml) was stirred for 1 hour at room temperature. The solvent was evaporated and the residue was dissolved in CH2Ck- NaOH (50%)was added and the mixture was extracted. The separated organic layer was dried(MgSO4), filtered and the solvent was evaporated in vacuum. The residue was purified by short column chromatography over silica gel (eluent: CH2CJ2/(MeOH/NH3) 100/0;95/5). The pure fractions were collected and the solvent was evaporated. Yield : 14.32g of intermediate 10 (59%). Example Al.k Preparation of intermediate 11 A mixture of intermediate 10 (0.00599 mol), l-Chloro-2-propanone (0.00599 mol) and K2CO3 (0.01199 mol) in DMF (200 ml) was stirred for 24 hours at room temperature. The solvent was evaporated. The residue was dissolved in CH2C12. The organic solution was washed with water, dried (MgSCU), filtered and the solvent was evaporated in vacuo. Yield: (quantitative yield) of intermediate 11 . Example A 1.1 Preparation of intermediate 12 Reaction under N2 atmosphere. A mixture of 3,4-Dihydro-2-naphthalenecarboxylic acid (0.0043 mol) and l,l'-carbonylbis[lH-imidazole] (0.0047 mol) in CH2C12) dry was stirred for one hour at room temperature. A solution of intermediate 7 (prepared according to Al.g) (0.0043 mol) in CH2C12, dry was added and the resulting reaction solution was stirred for ± 24 hours at room temperature. The solution was washed with water, then extracted with CH2C12. The separated organic layer was dried (Na2SO4), filtered and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2C12/CH3OH 97/3). The pure fractions were collected and the solvent was evaporated. Yield: 0.4 g of intermediate 12 (22%). Example Al.m Preparation of intermediate 13 Ethenyltriphenylphosphonium bromide (0.0025 mol) was added to a solution of intermediate 7 (prepared according Al.g) (0.003 mol) in CH2C12 (20 ml). The reaction mixture was stirred for 4 hours at room temperature. The solvent was evaporated under reduced pressure. Yield: 2.2 g of intermediate 13 , used in next reaction step, without further purification. Example Al.n Preparation of intermediate 14 To a solution of (E)-3-iodo-2-methylpropenoic acid (0.009 mol) in CH2Ch, dry (100ml) at room temperature underNj flow, l.l'-carbonylbisflH-imidazole] (0.0099 mol) was added. The mixture was stirred for 1 hour, then intermediate 7 (prepared according to Al.g) (0.009 mol) was added. The reaction mixture was stirred at room temperature for 16 hours, washed with H2O and brine, dried (Na2SCO4) and the solvent was evaporated under reduced pressure. The residue (white foam) was purified by short open column chromatography over silica gel (eluent: CPfcCh/MeOH 99/1). The pure fractions were collected and the solvent was evaporated. Yield : 3.82g of intermediate 14 (white solid, 81%). Example Al.o Preparation of intermediate 15 A solution of LiAlHU, 1.0 M/THF (0.00848 mol) in THF (100ml) was stirred and reffuxed under N2 flow at -20°C. A1C13 (0.0093 mol) was added in one portion and the resulting mixture was stirred at -20°C for 10 minutes. A solution of intermediate 14 (prepared according to Al.n) (0.0077 mol) in THF (100ml) was added dropwise and the resulting mixture was stirred at -20°C for 1 hour. A saturated NlitCl-solution 20% was added dropwise at -10°C and the reaction mixture was allowed to warm to room temperature. H2O was added to the suspension and was extracted with CH2Ch. The separated organic layer was dried (Na2SCO4) and the solvent was evaporated under reduced pressure. The residue was treated with Et2O and dried. Yield : 3.73g of intermediate 15 (white solid, 94%). Example Al.p Preparation of intermediate 16 A mixture of intermediate 7 (prepared according Al.g) (0.015 mol), l-chloro-2- propanone (0.015 mol) and K2CO3 (0.030 mol) in CH3CN (60 ml) was stirred for 24 hours at room temperature. The solvent was evaporated. The residue was partitioned between water and CHaCh. The separated organic layer was dried (NaaSOt), filtered and the solvent evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CHaCb/CEbOH 95/5). The pure fractions were collected and the solvent was evaporated. Yield: 4.79 g of intermediate 16 (82%). Example Al.q Preparation of intermediate 17 .0128 mol) was added portionwise to a solution of intermediate 16 (prepared according Al.p) (0.0051 mol) and H2O (3.2 ml) in THF (40.5 ml), at 0 °C. The reaction mixture was stirred overnight at room temperature, then treated with a 10% aqueous NHtCl solution. This mixture was extracted with CB2C2. The separated organic layer was dried, filtered and the solvent evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CKbCh/CH/jOH 97/3). The desired fractions were collected and the solvent was evaporated. Yield: 1.6 g of intermediate 17 (80%). Example Al.r Preparation of intermediate 18 Intermediate 7 (prepared according Al.g) (0.03 mol) was dissolved in CH3CN (200 ml) and K2COa (0.27 ml) was added. Oxiranemethanol (0.27 mol) was added and the reaction mixture was stirred over the weekend at 60°C. The solvent was evaporated in vacuo. The residue was partitioned between water and CH2C12. The organic layer was separated, dried (Na2SO4), filtered and the solvent was evaporated under reduced pressure. The residue was purified by preparatory HPLC ((1) eluent: CH2C12/(CH3OH/NH3) 95/5, then (2) eluent: CH2C12/CH3OH 90/10). The product fractions were collected and the solvent was evaporated. Yield: 7.5 g (61%) of pure intermediate 18 and 3.5 g of a mixture of starting material/target compound 1/1. Example Al.s Preparation of intermediate 19 Intermediate 18 (prepared according Al.r) (0.0012 mol) was dissolved in CH2Cl2 (20 ml). A solution of periodic acid sodium salt (0.0024 mol) in NaHCO3/H20 (q.s.) was added and the resulting reaction mixture was stirred vigorously for 2 hours. The mixture was partitioned between water and CH2O2. The separated organic layer was washed with brine, dried (Na2SCO4), filtered and the solvent evaporated in vacuo. Yield: 0.430 g of intermediate 19 (quantitative yield; used in next reaction step, without further purification). Example A2.a Preparation of intermediate 20 Reaction under N2 atmosphere. A solution of 2,2,2-trifluoro-N-(2-formylphenyl)- acetamide, (0.1869 mol) in DMF (375 ml) was added dropwise to NaH (0.2055 mol) in DMF (375 ml). The mixture was stirred for 30 min. at room temperature. A solution of 4-Bromo-3-butenoic acid methyl ester (0.2803 mol) in DMF (200 ml) was added dropwise. Then, 18-crown-6 (catalytic quantity) was added. The resulting reaction mixture was stirred for 2 hours at 60 °C, then overnight at room temperature. The solvent was evaporated. The residue was washed in water and extracted with CH2C12. The separated organic layer was dried (Na2SO4), filtered and the solvent evaporated. The residue was purified by open column chromatography over silica gel (eluent: CH2Cl2/hexane 90/10, 100/0 and with CH2Cl2/2-propanone 96/4, 90/10 and 80/20). The pure fractions were collected and the solvent was evaporated. Yielding: 44.37 g of intermediate 20 (75%, used in next reaction step, without further purification). Example A2.b Preparation of intermediate 21 Hydroxylamine (0.169 mol) and pyridine (0.211 mol) were added to a solution of intermediate 20 (prepared according to A2.a) (0.1407 mol) in ethanol (450 ml) and the resulting reaction mixture was stirred for 3 hours at room temperature. The mixture was washed with a 10% citric acid solution, then extracted with CH2Cl2. The separated organic layer was dried (Na2SCO4), filtered and the solvent was evaporated. Yielding: 45.76 g of intermediate 21 (98%, used in next reaction step, without further purification). Example A2.c Preparation of intermediate 22 or F A mixture of intermediate 21 (prepared according to A2.b) (0.0658 mol) and N-chloro- 4-methyl-benzenesulfonamide, sodium salt (0.0658 mol) in ethanol (500 ml) was stirred and refluxed for 2 hours. The mixture was concentrated in vacuo, filtered over dicalite, and the filtrate was washed with water and brine, then extracted with CH2C12. The separated organic layer was dried (Na2SO), filtered and the solvent evaporated. The residue was purified by open column chromatography over silica gel (eluent: CH2Cl2/2-propanone 100/0, 96/4, 90/10 and 80/20). The desired fractions were collected and the solvent was evaporated. The residue (syrup) was crystallized from hexane, then washed with DIPE, and dried. Yielding: 12.32 g of intermediate 22 (57%). Example A2.d Preparation of intermediate 23 NaBHt (0.0289 mol) was added portionwise to a mixture of intermediate 22 (prepared according to A2.c) (0.0116 mol) in THF (81 ml) and H2O (6.8 ml), stirred and cooled on an ice-bath. The resulting reaction mixture was stirred overnight at room temperature. The mixture was treated with a saturated aqueous NHtCl solution, then extracted with EtOAc. The separated organic layer was dried (Na2SCO4), filtered and the solvent was evaporated. The residue was washed with CH2C12, then recrystallized from EtOAc. The precipitate was filtered off and dried. Yielding: 0.9 g of intermediate 23 (38%). Example A2.e Preparation of intermediate 24 A mixture of intermediate 23 (prepared according A2.d) (0.001468 mol) and triphenylphosphine (0.001909 mol) in tetrachloromethane (30 ml) and THF (20 ml) was stirred and refluxed for 3 hours. The solvent was evaporated till dryness. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/hexane 90/10, then 100/0). The desired fractions were collected and the solvent was evaporated. The residue was crystallized from methanol. The precipitate was filtered off and dried. Yielding: 2.6 g of intermediate 24 (79%). Example A3.a Preparation of intermediate 25 I V 1,1-Dimethylethyl 1-piperazinecarboxylate (0.02 mol) was added portionwise to a solution of l,4-dichloro-2-butene (0.025 mol) in CHCls (60 ml). The reaction mixture was stirred for 24 hours at room temperature, then stirred and refluxed for 24 hours. The reaction was quenched with a saturated aqueous NaHCOs solution, dried (Na2SO4), filtered and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/EtOAc). The pure fractions were collected and the solvent was evaporated. Yielding: 2.2 g of intermediate 25 (40%). Example A3.b Preparation of intermediate 26 Reaction was carried out under Nj flow. A mixture of NaH, 60% (0.0579 mol) and 18- crown-6 (cat.quant.) in THF (25ml) was cooled. A mixture of 2-Amino-4,5- dimethoxybenzaldehyde (0.0579 mol) in THF (50ml) was added portionwise. The reaction was stirred at room temperature for 30 min. A mixture of intermediate 25 (prepared according to A3.a) (0.0386 mol) in THF (50ml) was added portionwise. The mixture was stirred at room temperature for 3 days and then treated with NBUC1 (10%). The mixture was extracted with CH2C12. The organic layer was separated, dried, filtered and the solvent was evaporated till dryness. The residue was purified by short open column chromatography (eluent: CH2C12/CH3OH 99/1 and 98/2). The pure fractions were collected and the solvent was evaporated. Yield: 5.5g of intermediate 26 (23%). Example A3.c Preparation of intermediate 27 The reaction was carried out under N2 flow. A mixture of intermediate 26 (prepared according A2.b) (0.02 mol) in THF (80ml) and 18-crown-6 (cat.quant.) were added portionwise to a mixture of NaH, 60% (0.03 mol) in THF (20ml). The mixture was stirred at room temperature for 20 min. trifluoroacetic acid anhydride (0.022 mol) was added portionwise. The mixture was stirred at room temperature for 3 hours, treated with a solution of NH4Cl (20%) and then extracted with CH2C12 and the solvent was evaporated till dryness. The residue was purified by short column chromatography over silica gel (CH2C12/CH3OH 99/1 and 98/2). The pure fractions were collected and the solvent was evaporated. Yield: 5.3g of intermediate 27 (58%). Example A3.d Preparation of intermediate 28 A mixture of intermediate 27 (prepared according to A3.c) (0.0115 mol), hydroxylamine (0.0126 mol) and NaHCO3 (0.023 mol) in ethanol, abs. (60ml) was stirred at room temperature for 24 hours, filtered off and the solvent was evaporated till dryness. Yield: 5.8g of intermediate 28 (95%). Example A3.e Preparation of intermediate 29 l-chloro-2,5-Pyrrolidinedione (0.0272 mol) was added portionwise to a solution of intermediate 28 (prepared according to A3.d) (0.0109 mol) in CH2C12 (100ml). The mixture was stirred at room temperature for 2 hours. Et3N (0.0272 mol) was added dropwise. The mixture was stirred at room temperature overnight, quenched with a K2CC3 10% solution, then extracted and the solvent was evaporated till dryness. Yield: of intermediate 29 . Example A3.f Preparation of intermediate 30 A mixture of intermediate 29 (prepared according to A3.e) (0.0109 mol) and LiOH (0.0119 mol) in H2O (17.5ml) and 1,4-dioxane (70ml) was stirred at room temperature for 3 hours. The mixture was treated with a NaOH (2N) solution and then extracted with CH2C12. The solvent was evaporated till dryness. The residue was purified by short open column chromatography (eluent: CH2Cl2/CH3OH 98/2). The pure fractions were collected and the solvent was evaporated. Yield: 2.07g of intermediate 30 (45%). Example A3.g N o Preparation of intermediate 31 Trifluoroacetic acid (7.9ml) was added dropwise to a solution of intermediate 30 (0.0047 mol) in CH2C12 (33ml). The mixture was stirred at room temperature for 3 hours, cooled and basified with a 50% NaOH solution. The mixture was extracted and the solvent was evaporated till dryness. Yield: 1.6g of compound 31 (100%). Example A4.a Preparation of intermediate 32 l,4-dichloro-2-butene (0.03 mol) was added to a mixture of l-(2- naphthylmethyl)piperazine (0.025 mol) and NaHCO3 (0.025 mol) in CH2C12 (75 ml). The reaction mixture was stirred for 24 hours at room temperature. The solid was filtered off, washed with more CH2C12 and the organic solution was washed with a 10% Na2CCO3 solution, dried (Na2SO4), filtered and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/EtOAc/2-propanone). The pure fractions were collected and the solvent was evaporated. Yield: 3.4 g of intermediate 32 (43%). Example A4.b Preparation of intermediate 33 A solution of NS (0.0546 mol) and teit-Butylamine (0.0983 mol) in toluene (75 ml) was stirred and refluxed for 24 hours using a Dean-Stark water separator. The solvent was evaporated. The residue was purified biyi.S-tiUatjjSW (bp at 0.5 mm Hg: 75 °C). Yielding: 8.1 g of intermediate 33 (72%)., Example A4.c Preparation of intermediate 34 Reaction under N2 atmosphere. n-BuLi (0.014 mol) was added dropwise to a solution of intermediate 33 (prepared according A4.a) (0.0125 mol) and 2,2,6,6- tetramethylpiperidine (0.0012 mol) in THF, dry (25 ml), stirred at -78 °C. The mixture was stirred for 3 hours at -10 °C. A solution of intermediate 32 (prepared according A4.b) (0.0083 mol) in THF, dry (25 ml) was added portionwise at -10 °C. The reaction mixture was stirred for 24 hours at room temperature, then quenched with NHjCl (10%) and the organic layer was separated. The aqueous layer was extracted with CH2C12. The combined organic layers were dried (Na2SO4), filtered and the solvent was evaporated. Yielding: 5.6 g of intermediate 34 (100%) Example A4.d Preparation of intermediate 35 NaHCO3 (0.015 mol) and hydroxylamine (0.0125 mol) were added to a solution of intermediate 34 (prepared according A4.c) (0.0083 mol) in ethanol, abs. (50 ml). The reaction mixture was stirred for 24 hours at room temperature. CHaC^ was added and the solid was filtered off and washed with CH2Cl2. The solvent was evaporated. The residue was taken up into QHCh and washed with 10% Na2CO3 and with brine. The organic layer was separated, dried (Na2SCO4.), filtered and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/EtOAc/2-propanone). The desired fractions were collected and the solvent was evaporated. Yielding: 0.9 g of intermediate 35 (24%). Example A5.a -N-^OO-b Preparation of compound 36 A mixture of A (0.029 mol) and intermediates (prepared according to Al.e) (0.0058 mol) in 1,4-dioxane (5 ml) was stirred for 6 hours at 100 °C. The solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2C12/CH3OH 98/2 - 97/3). The product fractions were collected and the solvent was evaporated. Yield: 3.3 g of intermediate 36 Example A5.b (Figure Removed) Preparation of compound 37 ll if II Trifluoroacetic acid (11.7 ml) was added dropwise to a solution of intermediate 36 (prepared according to A5.f) (0.0071 mol) in CHC13 (50 ml) and the resulting reaction mixture was stirred for 3 hours at ± 10 °C. The reaction mixture was cooled, then further alkalized with 50% NaOH. This mixture was extracted and the extract's solvent was evaporated. Yield: 2.5 g of intermediate 37 (quantitative yield; used in next reaction step, without further purification). B. Preparation of the final compounds Example Bl. a Preparation of compound 1 A mixture of intermediates (prepared according to Al.e) (0.0291 mol) and l-(3- phenyl-2-propenyl)-piperazine, (0.0582 mol) was heated for 2 hours at 100 °C. The crude reaction mixture was washed with water and extracted with CH2C12. The separated organic layer was dried (Na2SCO4), filtered and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2C12/CH3OH 95/5) and by HPLC over silica gel (eluent: CH2C12/CH3OH 80/20). The pure fractions were collected and the solvent was evaporated. This fraction was separated into its optical enantiomers by chiral column chromatography over Chiralpak AD (eluent: C2H5OH/CH3CN 90/10). The (B)-enantiomeric fractions were collected and the solvent was evaporated. The residue was dissolved in methanol and converted into the hydrochloric acid salt (1:2). The precipitate was filtered off and dried. Yielding: 2.47 g of compound 1 . ExampleBl.b Preparation of compound 2 A mixture of intermediate 5 (prepared according to Al.e) (0.0044 mol) and (3-phenyl- 2-propenyl)-piperazine (0.0087 mol) was stirred for 2 hours at 100 °C. The reaction mixture was purified by short open column chromatography over silica gel (eluent: CH2C12/CH3OH 95/5), then by high-performance liquid chromatography over silica gel (eluent: CH2Cl2/CH3OH 96/4). The pure fractions were collected and the solvent was evaporated. The residue (1.4 g) was treated with diethyl ether, then dried. Yielding: 1.2 g of compound 2 (60%). Example Bl.c Preparation of compound 3 A mixture of intermediate 6 (0.00227 mol), (E) l-(2~methyl-3-phenyl-2- propenyl)piperazine (0.00273 mol) and NaHCO3 (0.00455 mol) in dioxane (30 ml) was stirred and refluxed for 48 hours. The solvent was evaporated. The residue was dissolved in CH2C12. The organic solution was washed with water, dried (MgSCU), filtered and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2C12/(CH3OH/NH3) 99/1), then by HPLC (eluent: CH2C12/CH3OH 99.5/0.5 to 98/2). The desired fractions were collected and the solvent was evaporated. Yield: 0.17 g of compound 3 . Example Bl.d Preparation of compound 4 B-CIS A mixture of intermediate 8 (prepared according to Al.h) (0.0058 mol) and HNx jt (0.0116 mol) in dioxane (10ml) was stirred and refluxed for 8 hours, then stirred overnight at room temperature, then stirred and refluxed for 18 hours. The mixture was treated with H2O and extracted with CH2Cl2- The solvent of the separated organic layer was evaporated. The residue was purified by short open column chromatography (eluent: CHzClj/MeOH 97/3). The desired fractions were collected and the solvent was evaporated. The residue was treated with diethyl ether, then dried. Yield : 0.9g of compound 4 (33%). Example Bl.e Preparation of compound 5 A mixture of intermediate 10 (prepared according to Al.i) (0.0029 mol), (0.0058 mol), AcOH (0.48g) and (AcO)3BHNa (0.4g) in 1,2-dichloroethane (20ml) was stirred and refluxed overnight. The mixture was treated with HaO and extracted. The solvent of the separated organic layer was evaporated. The residue was purified by short column chromatography over silica gel (eluent : CHaC12/MeOH 97/3). The desired fractions were collected and the solvent was evaporated. The residue was treated with diethyl ether, then dried. Yield : l.OTg of compounds (82%). Example Bl.f Preparation of compound 6 A mixture of compound 3 (prepared according ex. Bl) (0.00020 mol), acetylchloride (0.00024 mol) and Et3N (0.00061 mol) in chloroform (10 ml) was stirred at room temperature for 2 hours. Water was added and this mixture was extracted with CH2Cb- The separated organic layer was dried (MgSCU), filtered and the solvent evaporated in vacuo. The residue was purified by CC-TLC on Chromatotron (eluent: CH2C12/CH3OH 97/3; then 99/1). The desired fractions were collected and the solvent was evaporated. Yield: 0.022 g of compound 6 . Example Bl.g Preparation of compound 7 Preparation of compound 8 Compound 2 (prepared according Bl.b) (0.0022 mol) was separated and purified into its optical enantiomers by chiral column chromatography over Chiralpak AD (eluent: C2HsOH/CH3CN 90/10). Two fraction groups were collected and their solvent was evaporated. Yielding: ± 1.5 g of fraction 1 (LCI purity: > 99.5%) and ± 1.5 g of fraction 2 (LCI purity: > 99.5%). Fraction 1 was crystallized by treatment with hexane, stirring overnight. The precipitate was filtered off and dried. Yielding: 1.08 g of compound compound 7 (grease solid). Fraction 2 was crystallized by treatment with EtOAc, stirring overnight. The precipitate was filtered off and dried. Yielding: 0.54 g of 8 (grease solid). Example B2.a Preparation of compound 9 A mixture of intermediate 24 (prepared according A2.e) (0.0022 mol), l-(2- naphthalenylmethyO-piperazine, (0.0044 mol) and KI (catalytic quantity) in 1,4-dioxane (2.5 ml) was stirred and refluxed overnight. The reaction mixture was washed with water and this mixture was extracted with CH2C The residue was purified by short open column chromatography over silica gel (eluent: C2CWCHsOH 98/2), then by HPLC (eluent: CH2Cl2/(CH3OH/NH3) 96/4). The pure fractions were collected and the solvent was evaporated. The residue was treated with DIPE, filtered off and dried. Yield: 0.3 g of compound 9 (30%). Example B2.b Preparation of compound 10 Reaction under N2 atmosphere. A solution of compound 9 (prepared according to B2.a) (0.0012 mol) in THF, dry (3 ml) and 18-crown-6 (catalytic quantity) was slowly added to a solution of NaH, 60% (0.0018 mol) in THF, dry (2 ml). The reaction mixture was stirred for 30 min at room temperature, acetylchloride (0.0013 mol) was added dropwise and the reaction mixture was stirred for 3 hours at room temperature. The reaction mixture was treated with aqueous NH^Cl and extracted with CH2C12. The separated organic layer was dried, filtered and the solvent evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2C12/CH3OH 98/2), then by HPLC (eluent: CH2C12/(CH3OH/NH3) 98/2). The pure fractions were collected and the solvent was evaporated. Yield: 0.26 g of compound 10 (52%). Example B3a Preparation of compound 11 A mixture of intermediate 31 (prepared according A3.g) (0.0045 mol), (E)- (3-chloro-2-methyl-l-propenyl)-benzene (0.0037 mol) and K2CO3 (0.0037 mol) in DMF (15 ml) was stirred at 70°C for 2 hours. The mixture was washed with water and then extracted with CH2Cl2 . The organic layer was separated, dried, filtered and the solvent was evaporated. The residue was purified by short column chromatography over silica gel (eluent: CH2C12/CH3OH 98/2). The desired fractions were collected and the solvent was evaporated. The residue was purified again by HPLC (eluent: CH2Cl2/(CH3OH/NH3) 98/2). The pure fractions were collected and the solvent was evaporated. The residue was treated with DBPE. The precipitate was filtered off and dried. Yield: 0.34 g of compound 11 (20%). Example B3.b Preparation of compound 12 Preparation of compound 13 Compound 11 (prepared according to B3.a) (0.00605 mol) was purified by highperformance liquid chromatography over Chiralcel OJ (eluent: hexane/MeOH/EtOH 20/24/56). The desired fractions were collected and the solvent was evaporated. Yield : fractions A and B. Fraction A was purified by high-performance liquid chromatography over RP BBS C18 (eluent: (0.5%NH4OAc in H2O/CH3CN(90/10))/MeOH 70/30). The pure fractions were collected and the organic solvent was evaporated. The aqueous layer was extracted with CH2C12. The separated organic layer was dried (MgSO4), filtered and the solvent was evaporated. The residue was stirred in hexane and the precipitate was filtered off. Yield : 0.69g of compound 12 . Fraction B was purified by high-performance liquid chromatography over RP BDS CIS (eluent: (0.5%NH4OAc in H2O/CH3CN(90/10))/MeOH 70/30). The pure fractions were collected and the organic solvent was evaporated. The aqueous layer was extracted with CH^Ch. The separated organic layer was dried (MgSCU), filtered and the solvent was evaporated. The residue was stirred in hexane and the precipitate was filtered off. Yield : 0.67g of compound 13 . Example 4 Preparation of compound 14 Reaction done in solid phase using a Quest 210 synthesizer (Argonaut Technologies, San Carlos, USA). N,N-Diisopropylethylamine (0.0036 mol) was added to a suspension of (°-0006 mo1)in acetonitrile (4 ml). l-(2-Chlorophenylmethyl)piperazine (0.0012 mol) was added and the resulting reaction mixture was stirred for 20 hours at 80 °C. Then, each reaction vessel was filtrated and the filtrate was evaporated. The residue was HPLC purified. The pure fractions were collected and the solvent was evaporated. Yield: 0.102 g of compound 14 Example B5.a Preparation of compound 15 A mixture of intermediate 7 (prepared according Al.g) (0.0036 mol), 2-(bromomethyl)naphthalene (0.0055 mol) and K2CO3 (0.0055 mol) in MIK (15 ml) was stirred for ± 24 hours at 100 °C. The crude reaction mixture was washed with water, then extracted with CH2Cl2. The separated organic layer was dried (Na2SCO4), filtered and the solvent was evaporated. The residue was purified by high-performance liquid chromatography over silica gel (2 x) ((I) eluent: CH2C12/CH3OH 95/5; (II) eluent: CH2C12/(CH3OH/NH3) 98/2). The pure fractions were collected and the solvent was evaporated. Yielding: 0.2 g of compound 15 (11%). ExampleBS.b Preparation of compound 16 Preparation of compound 17 Compound 15 (prepared according B5.a) (0.0106 mol) was separated into its enantiomers by column chromatography (eluent: hexaneAI^HsOH gradient 30/70 to 0/100; column: CfflRALPAK AD 1000 A 20 /Am DIACEL). Two pure fractions were collected and their solvents were evaporated. The residue was dissolved in CH3OH and converted into the hydrochloric acid salt (1:2). The precipitate was filtered off and dried. Yielding: 2.08g of compound 16 (36%) and 2.19g of compound 17 (38%). Example B6 Preparation of compound 18 A mixture of intermediate 7 (prepared' according to Al.g) (0.0045 mol), 2-methyl-3-(3-thienyl)-2-propenal (0.00675 mol), NaBH(OAc)3 (0.00675 mol) and HO Ac (2 drops) in 1,2-dichloroethaan (30 ml) was stirred overnight at room temperature. A saturated aqueous NHtCl solution was added and this mixture was extracted with CHaCh. The separated organic layer was dried (MgSCO4), filtered and the solvent evaporated in vacuo. The residue was purified by flash column chromatography over silica gel (eluent: CH2C12/(CH3OH/NH3) 97/3), then by HPLC (eluent: CH2Cl2/(CH3OH/NH3) 99/1 to 98/2). The product fractions were collected and the solvent was evaporated. Yield: 0.965 g of compound 18 (46%; containing also 3% of the (Z) isomer!). Example B7 Preparation of compound 19 A mixture of intermediate 10 (prepared according to Al.j) (0.003 mol), 4-chlorobenzaldehyde (0.0045 mol) and (AcO)3BHNa (0.0045 mol) in 1,2-dichloroethane (30ml) was stirred and refluxed for 2 hours at room temperature. A saturated aqueous NEUCl-solution was added and the mixture was extracted with CH2C12. The separated organic layer was dried (MgSO4), filtered and the solvent was evaporated in vacuum. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/(MeOH/NH3) 97/3). The desired fractions were collected and the solvent was evaporated. The residue was precipitated from DIPE. Yield: 1.180g of compound 19 (57%). Example B8 Preparation of compound 20 A mixture of (0.00036 mol), 2-methylbenzaldehyde (0.00108 mol) and NaOCH3, 30% in CH3OH (0.00108 mol) in CH3OH, dry (8 ml) was stirred for 16 hours at 65 °C (Reaction done in solid phase using a Quest 210 synthesizer (Argonaut Technologies, San Carlos, USA)). The mixtures were filtered and the filtrate was HPLC purified (eluent: CH2Cl2/(CH3OH/NH3) 98/2). The desired fractions were collected and the solvent was evaporated. Yield: 0.032 g of compound 20. Example B9 Preparation of compound 21 Reaction under N2 atmosphere. Solution LiAIKU, 1 M/THF (0.8 ml) was stirred at -20 °C. A1C13 (0.0009 mol) was added in one portion. The resulting solution was stirred for 10 min at -20 °C. A solution of intermediate 12 (prepared according to Al.l) (0.0008 mol) in THF, dry (5 ml) was added dropwise and the resulting reaction mixture was stirred for one hour at -20 °C. Then; a saturated aqueous NH4Cl solution was added carefully. The reaction mixture was washed with water, then extracted with CH2C12. The separated organic layer was dried (Na2SCO4), filtered and the solvent was evaporated. The residue was treated with ether. The residue (0.13 g) was purified by CC-TLC Chromatotron (eluent: CH2C12/CH3OH 97/3). The pure fractions were collected and the solvent was evaporated. Yield: 0.09 g of compound 21 (30%). Example BIO Preparation of compound 22 Reaction under N2 flow. A mixture of intermediate 13 (prepared according to Al.m) (0.001 mol) in CH3OH, dry (20 ml) was stirred at room temperature. NaOCH3, 30% in CH3OH (0.002 mol) was added. 5-Indanecarboxaldehyde (0.002 mol) was added and the resulting reaction mixture was stirred and refluxed for 16 hours. The reaction mixture was allowed to cool to room temperature. 20% NH4Cl was added and this mixture was extracted with CH2Cl2. The separated organic layer was washed with water, with brine, dried (NajSO4), filtered, and the solvent was evaporated under reduced pressure. The residue was purified by short open column chromatography and CC-TLC (eluent: CH2C12/CH3OH 98/2). The product fractions were collected and the solvent was evaporated. Yield: 0.035 g of compound 22 (7.2%, light-brown solid). Example B11 Preparation of compound 23 A mixture of intermediate 15 (prepared according to Al.o) (0.00136 mol), 2- (trimethylstannyl)pyridine (0.0027 mol) and Pd(PPh3)4 (0.00013 mol) in toluene (20ml) was heated to 100°C. The reaction mixture was stirred for 16 hours and was allowed to cool to room temperature. H2O was added and the mixture was extracted with CH2C12. The separated organic layer was collected, washed with H2O and brine, dried (Na2SO4) and the solvent was evaporated under reduced pressure. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/MeOH 98/2). The pure fraction was collected and the solvent was evaporated. The resulting residue was purified by CC-TLC on Chromatotron (eluent: CH2CL2/MeOH 98/2). The pure fraction was collected and the solvent was evaporated. Yield : 0.044g of compound 23 (light brown solid, 7%). ExampleB12 Preparation of compound 24 Reaction under N2 atmosphere. n-BuLi, 2.5M/hexanes (0.0062 mol) was added dropwise to a stirred solution of (p-Flurorobenzyl)triphenylphosphonium chloride (0.0062 mol) in THF (20 ml). The mixture was stirred for 15 min. A solution of intermediate 11 (prepared according Al.k) (0.00514 mol) in THF (20 ml) was added dropwise. The reaction mixture was stirred for 16 hours at 50 °C. Water was added and this mixture was extracted with Et2O. The separated organic layer was dried (MgSCU), filtered and the solvent evaporated in vacuo. The residue was purified by short open column chromatography over silica gel (eluent: CHkCh/CHsOH 97/3), then by HPLC (eluent: CH2Cl2/(CH3OH/NH3) 99/1) to separate the (E)/(Z) isomers. Two product fraction groups were collected and their solvent was evaporated. Yield: 0.651 g of compound 24 (26%, (E)). ExampleB 13 Preparation of compound 25 Resin f (0.0016 mol; 1.5 mmole/g) was suspended in THF. 1.6 M BuLi (0.0015 mol) was added and the mixture was stirred for 15 min. The mixture was filtered and the filter residue (resin) was washed with anhydrous THF (3 x). The resin was suspended in THF (5 ml). Intermediate 19 (prepared according to Al.s) (0.0004 mol) was added and the reaction mixture was stirred overnight at 100 °C. The mixture was cooled, filtered and the filtrate was evaporated in vacuo. The residue was purified by preparatory HPLC (eluent: CH2Cl2/(CH3OH/NH3) 97/3). The product fractions were collected and the solvent was evaporated. Yield: 0.168 g of compound 25 . Example B14 Preparation of compound 26 A mixture of intermediate 16 (prepared according to Al.p) (0.0025 mol), benzenamine (0.0028 mol) and NaBH, (0.0028 mol) in HOAc (50 ml) was stirred for 2 hours at room temperature. An aqueous NHiOH solution was added. This mixture was extracted with CH2Ch. The separated organic layer was dried (MgSO4), filtered and the solvent evaporated in vacuo. The residue was purified by short open column chromatography over silica gel (eluent: CH2Cl2/(CH3OH/NH3) 99/1), then by flash column chromatography over silica gel (eluent: CH2Cl2/(CH3OH/NH3) 98/2). The product fractions were collected and the solvent was evaporated. Yield: 0.181 g of compound 26 (15%). Example B15 Preparation of compound 27 (0.0018 mol) was added dropwise to a solution of intermediate 17 (prepared according to Al.q) (0.0012 mol), 3-fluorophenol (0.0018 mol) and PPh3, pol. (0.0024 mol) in THF, dry (10 ml), under N2 atmosphere. The reaction mixture was stirred overnight at room temperature. The mixture was filtered, washed with CH^Ch and CH3OH, and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2C12/EtOAc 2/1 and 96/4), then by flash column chromatography over silica gel (eluent: 97/3). The product fractions were collected and the solvent was evaporated. Yield: 0.29 g of compound 27 (50%). Example B16 Preparation of compound 28 NaClO (4%) (0.005 mol) was added to a solution of intermediate 35 (prepared according A4.d) (0.002 mol) in CH2C]2 (10 ml). The reaction mixture was stirred for 4 hours at room temperature. Et3N (0.004 mol) was added and the reaction mixture was stirred for 24 hours at room temperature. The organic layer was separated, washed with Na2SCO3 (10%), dried (Na2SCU), filtered and the solvent was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: EtOAc and CH2Cl2/CH3OH 95/5). The desired fractions were collected and the solvent was evaporated. The residue was taken up into diethyl ether, then filtered through dicalite and the filtrate was evaporated. The residue was dissolved in diethyl ether and converted into the hydrochloric acid salt (1:2). The precipitate was filtered off, washed with 2-propanone and diethyl ether, and dried. Yield: 0.15 g of compound 28 (15%). ExampleB 17 Preparation of compound 29 A mixture of intermediate 37 (prepared according to A5.g) (0.006 mol) and 2- (bromomethyl)naphthalene (0.003 mol) in dioxane (40 ml) was stirred at 100 °C for 6 hours, then overnight at room temperature. The reaction mixture was treated with a 10% aqueous K2CO3 solution, then extracted with CHjC^. The separated organic layer was evaporated. The residue was purified by short open column chromatography over silica gel (eluent: CH2CI2/CH3OH 95/5 and 90/10 and CH2Cl2/(CH3OH/NH3) 95/5). The product fractions were collected and the solvent was evaporated. The residue (1.49 g) was treated with diethyl ether, then dried. Yield: 0.8 g of compound 29 (53%). In the following tables (Tables 1-5) a number of compounds are given which have been prepared according to any one of the Examples above. All compounds have also been tested for their pharmacological activity. (Table Removed) C. Pharmacological examples Example Cl : Binding experiment for qadrenergic receptor subtypes and for 5-HT transporter General The interaction of the compounds of Formula (I) with ha2-receptors and H5-HTtransporters was assessed in in vitro radioligand binding experiments. In general, a low concentration of a radioligand with a high binding affinity for a particular receptor or transporter is incubated with a sample of a tissue preparation enriched in a particular receptor or transporter or with a preparation of cells expressing cloned human receptors in a buffered medium. During the incubation, the radioligand binds to the receptor or transporter. When equilibrium of binding is reached, the receptor bound radioactivity is separated from the non-bound radioactivity, and the receptor- or transporter-bound activity is counted. The interaction of the test compounds with the receptor is assessed in competition binding experiments. Various concentrations of the test compound are added to the incubation mixture containing the receptor- or transporter preparation and the radioligand. The test compound in proportion to its binding affinity and its concentration inhibits binding of the radioligand. The radioligand used for hcx2A> hcc2B and ha-2C receptor binding was [3Hj-raulwolscine and for the h5-HT transporter was [3H]paroxetine. Cell culture and membrane preparation. CHO cells, stabile transfected with human adrenergic-cx2A-, -WIB or OL-IC receptor cDNA, were cultured in Dulbecco's Modified Eagle's Medium (DMEM)/Nutrient mixture Ham's F12 (ratio l:l)(Gibco, Gent-Belgium) supplemented with 10 % heat inactivated fetal calf serum (Life Technologies, Merelbeke-Belgium) and antibiotics (100 lU/ml penicillin G, 100 /ig/ml streptomycin sulphate, 110 g/ml pyruvic acid and 100 jig/ml L-glutamine). One day before collection, cells were induced with 5 mM sodiumbutyrate. Upon 80-90 % of confluence, cells were scraped in phosphate buffered saline without Ca2"1" and Mg2+ and collected by centrifugation at 1500 x g for 10 min. The cells were homogenised in Tris-HCl 50 mM using an Ultraturrax homogenizer and centrifuged for 10 min at 23,500 x g. The pellet was washed once by resuspension and rehomogenization and the final pellet was resuspended in Tris-HCl, divided in 1 ml aliquots and stored at -70°C. Binding experiment for cfe-adrenergic receptor subtypes Membranes were thawed and re-homogenized in incubation buffer (glycylglycine 25 mM, pH 8.0). In a total volume of 500 n\, 2-10 /ig protein was incubated with [3H]raulwolscine (NET-722) (New England Nuclear, USA) (1 nM final concentration) with or without competitor for 60 min at 25°C followed by rapid filtration over GF/B filter using a Filtermatel96 harvester (Packard, Meriden, CT). Filters were rinsed extensively with ice-cold rinsing buffer (Tris-HCl 50 mM pH 7.4). Filter-bound radioactivity was determined by scintillation counting in a Topcount (Packard, Meriden, CT) and results were expressed as counts per minute (cpm). Non-specific binding was determined in the presence of 1 M oxymetazoline for h(X2A- and receptors and 1 /iM spiroxatrine for hccac receptors. Binding experiment for 5-HT transporter Human platelet membranes (Oceanix Biosciences Corporation, Hanover, MD, USA) were thawed, diluted in buffer (Tris-HCl 50 mM, 120 mM NaCl and 5 mM KC1) and quickly (max 3 s) homogenised with an Ultraturrax homogenizer. In a total volume of 250 uL, 50-100 ug protein was incubated with [3H]paroxetine (NET-869) (New England Nuclear, USA) (0.5 nM final concentration) with or without competitor for 60 min at 25 °C . Incubation was stopped by rapid filtration of the incubation mixture over GF/B filters, pre-wetted with 0.1 % polyethyleneamine, using a Filtermatel96 harvester (Packard, Meriden, CT). Filters were rinsed extensively with ice-cold buffer and radioactivity on the filters was counted in a Topcount liquid scintillation counter (Packard, Meriden, CT). Data were expressed as cpm. Imipramine (at 1 p.M final concentration) was used to determine the non-specific binding. Data analysis and results Data from assays in the presence of compound were calculated as a percentage of total binding measured in the absence of test compound. Inhibition curves, plotting percent of total binding versus the log value of the concentration of the test compound, were automatically generated, and sigmoidal inhibition curves were fitted using non-linear regression. The pICso values of test compounds were derived from individual curves. All compounds according to formula (I) produced an inhibition at least at the htt2A site (but often also at the htt2B and ha2C sites) and simultaneously at the 5-HT transporter site of more than 50 % (pICso) at a test concentration ranging between 1(H-6 M and 10~9 M in a concentration-dependent manner. For a selected number of compounds, covering most of the various embodiments of Formula (I), the results of the in vitro studies are given in Table 6. Table 6 : Some results of the in vitro experiments (pICso-values). n.d.: not determined. Comp (Table Removed) Example C2 : In vivo experiment for O-adrenoceptor antagonism and for serotonine (5-HT) reuptake inhibition activity. A. Medetomidine-test The onset and end of medetomidine (0.10 mg/kg, i.v.)-induced loss of righting was recorded in overnight-starved Wiga male rats (200-250 g). The intensity of the loss of righting was scored: 0 = normal behaviour, 1 = slight ataxia, 2 = pronounced ataxia, 3 = loss of righting for a period 5 min. Under standard conditions, test compound or solvent was administered (s.c. or p.o.) 1 h before medetomidine. Criterion for drug-induced antagonism: (1) antagonism of loss of righting: duration = 0 min (1.4% false positive controls; n = 74) (2) reversal of ataxia: score righting reflex over a period longer than 120 min (0% false positives). Centrally acting tti-adrenoceptor antagonists or behavioural stimulants antagonise the loss of righting ; sedative compounds may result in prolongation. The following observations were made : onset of loss of righting (min), end of loss of righting (min) and intensity of loss of righting (score 0-4). The observations were performed at 1 h following s.c. (solutions) or p.o. (suspensions) administration, respectively. Starting dose was 10 mg/kg (References : Berger, U.V., Grzanna, R., Molliver, M.E., Exp. Neurol. 103, 111-115 (1989), Fuller, R.W., Perry, K.W., Molloy, B.B.. Eur. J. Pharmacol. 33, 119-124 (1975) and Lassen, B.J., Psvchopharmacol. 57, 151-153(1978)). B. pCA-test Male Wiga rats were used (body weight: 200 ± 20 g). One hour after administration of test compound or solvent, a solution of pCA was injected subcutaneously (5 mg/kg ; 10 ml/kg). Forty-five minutes after the pCA injection, head-twitches (HTW) are counted and the excitation (EXC) were scored over three successive 5 min intervals (starting 45, 50 and 55 minutes after pCA-administration. The scores were given by a trained observator according to the intensity scale: 0 = absent or doubtful, 1 = present, 2 = pronounced, 3 = maximal. For statistical analysis, the head-twitches counted during the 15-min observation time were cumulated. For the other phenomena, the median value of the three 5-min-intervals was used. Standard observations were performed at 1 h following s.c.or p.o. administration. The starting dose was in general 10 mg/kg. Doses were initially given to 2 animals. When both animals show activity for at least one of the observations, the compound was considered active and testing was repeated at a 4 times lower dose. When activity was found in only one out of the two animals, an additional animal was tested. When activity was found in this additional animal, the compound was also considered to be active and testing was repeated at a 4 times lower dose. In all other cases the compound was considered inactive at the particular time-route-dose (Reference : Janssen, P.A.J., Niemegeers, C.J.E., Awouters, F., Schellekens, K.H.L., Megens, A.A.H.P., Meert, T.FJ. Pharmacol. Exp. Thorn. 244. 685-693 (1988)). Results A large number of compounds according to the invention showed a central activity (minimal effective dose) both in the medetomidine test and in the pCA-test of less than or equal to 10 mg/kg. Example C3 : r3SS1GTPyS binding assay Membranes of the hccaA adrenoceptor expressing CHO cell line were thawed and rehomogenised in 20 mM Hepes buffer. The incubation medium consisted of: 20 mM Hepes buffer, pH 7.5,1 jiM GDP, 3 mM MgCl2, 100 mM NaCl, 0.25 nM [35S]GTPyS and 10 u.g protein per well of a 96-well plate. Antagonists and the reference agonist noradrenaline (3 pM) were added 20 min before the[35S]GTPyS. The incubation (20 min, 37°C) was ended by rapid filtration through GF/B filters and binding quantified by liquid scintillation counting. Results All compounds according to the invention evaluated in the GTPyS binding assay did not show significant increases of [35S]GTPyS binding to the ha2A receptor up to 10 uM. All compounds evaluated in the assay were able to inhibit noradrenaline-induced increases of [35S]GTPyS binding, thereby showing their antagonistic nature at this receptor. We claim 1. Isoxazoline compounds of general Formula (I) (Formula Removed) the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the N-oxide form thereof, wherein : X is CH2, N-R7, S or O; R7 is selected from the group of hydrogen, alkyl, phenyl, phenylalkyl, alkylcarbonyl, alkyloxycarbonyl and mono- and dialkylaminocarbonyl, the phenyl and alkyl groups being optionally substituted with one or more halo atoms; R1 and R2 are each, independently from each other, selected from the group of hydrogen, hydroxy, cyano, halo, OSO2H, OSO2CH3, phenyl, phenylalkyl, alkyloxy, alkyloxyalkyloxy, alkyloxyalkyloxyalkyloxy, tetrahydrofuranyloxy, alkylcarbonyl-oxy, alkylthio, alkyloxyalkylcarbonyloxy, pyridinylcarbonyloxy, alkylcarbonyloxyalkyloxy, alkyloxycarbonyloxy, alkenyloxy, alkenylcarbonyloxy and mono-and dialkylaminoalkyloxy, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; or R1 and R2 may be taken together to form a bivalent radical -R1-R2- selected from the group of -CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-O-, -CH2-O-CH2- and -O-CH2-CH2-O- ; a and b are asymmetric centers ; (CH2)m is a straight hydrocarbon chain of m carbon atoms, m being an integer ranging from 1 to 4 ; Pir is a radical according to any one of Formula (IIa), (IIB or (IIc) (Formula Removed) wherein : each R8 is independently from each other, selected from the group of hydrogen, hydroxy, amino, nitro, cyano, halo and alkyl ; n is an integer ranging from 1 to 5 ; R9 is selected from the group of hydrogen, alkyl and formyl; R3 is a radical according to any one of Formula (III) (Formula Removed) wherein : d is a single bond while Z is a bivalent radical selected from the group of -CH2-, -C(=O)-, -CH(OH)-, -C(=N-OH)-, -CH(alkyl)-, -O-, -S-, -S(=O), -NH- and -SH-; or d is a double bond while Z is a trivalent radical of formula =CH- or =C(alkyl); A is a 5- or 6-membered aromatic homocyclic or heterocyclic ring, selected from the group of phenyl, pyranyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl, oxadiazolyl and isoxazolyl; p is an integer ranging from 0 to 4 ; q is an integer ranging from 0 to 7 ; R4 is selected from the group of hydrogen, alkyl, phenyl, biphenyl, naphthyl, halo and cyano, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; R5 is equal to R4 ; or R4 and R5 may be taken together to form a bivalent radical -R4- R5- selected from the group of -CH2-, =CH,-CH2-CH2,-CH=CH-,-O-,-NH-,=N-,-S-,-CH2N (-alkyl, -CH=N -CH2O- and -OCH2- ; each R6 is independently from each other, selected from the group of hydrogen, hydroxy, amino, nitro, cyano, halo, carboxyl, alkyl, phenyl, alkyloxy, phenyloxy, alkylcarbonyloxy, alkyloxycarbonyl, alkylthio, mono- and dialkylamino, alkylcarbonylamino, mono- and dialkylaminocarbonyl, mono- and dialkyl-aminocarbonyloxy, mono- and dialkyl-aminoalkyloxy, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; or two vicinal radicals R6 may be taken together to form a bivalent radical -R6-R6- selected from the group of -CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-C(=O),-O- CH2-O-, -CH2 -O- CH2-,-O-CH2 -CH2 -0-, -CH=CH-CH=CH-,-CH=CH-CH=N-, -CH=CH- N=CH-,-CH= N-CH=CH-, -N=CH-CH= CH-, -CH2-CH2-CH2-, -CH2-CH2-C(=O)- and -CH2-CH2-CH2-CH2- ; and R10 is selected from the group of hydrogen, alkyl, phenylalkyl and phenyl. 2. A compound as claimed in claim 1, wherein X=O or NH ; R1 and R2 are both alkyloxy ; m = 1 ; Pir is a radical according to Formula (IIa) wherein R8 is hydrogen and n = 4 ; R3 is a radical according to Formula (IIIb) wherein Z is =CH-, d is a double bond, A is a phenyl ring, R4 is an alkyl and R10 is hydrogen . 3. A compound as claimed in any one of claims 1-2 for use as a medicine. 4. A prodrug of a compound of Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the iV-oxide form thereof, which is degraded in vivo to yield a compound according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the iV-oxide form thereof. 5. A process for preparing a compound as claimed in any one of claims 1-2, wherein a compound according to Formula (IV) is reacted with a compound according to Formula (V) or Formula (VII). 6. A compound of general Formula (VIII) (Formula Removed) wherein R1, R2, X, m, R8 and n are defined as in Formula (I).

Full Text

The present invention relates to isoxazoline compounds.
The invention concerns substituted isoxazolines derivatives having anti-depressant
activity and/or anxiolytic activity and/or body weight control activity, processes for
their preparation, pharmaceutical compositions comprising them and their use as a
medicine, in particular for the treatment of depression, anxiety, stress-related disorders
associated with depression and/or anxiety and disorders of body weight including
anorexia nervosa and bulimia.
The invention also relates to novel combination of substituted isoxazolines derivatives
having anti-depressant activity and/or anxiolytic activity and/or body weight control
activity with, anti depressants, anxiolytics and/or antipsychotics.
Tetrahydronaphtalene and indane derivatives showing anti-depressant activity are
known from EP-361 577 Bl. These compounds are typical monoamine reuptake
blockers with additional (X2-adrenoceptor antagonist activity and they show antidepressant
activity without being sedative.
The problems associated with the compounds according to the state of the art is that the
compounds cause considerable side-effects, such as nausea, excitation, an increased
heart rate and a reduced sexual function. Furthermore, it requires a long time, in
particular 3-4 weeks, before the response starts.
The purpose of the present invention is to provide novel compounds derivatives having
anti-depressant and/or anxiolytic and/or body weight control activity, in particular
compounds that do not exhibit the aforementioned disadvantages.
The present invention relates to novel isoxazoline derivatives according to the general
(Formula Removed)
the pharmaceuticalJy acceptable acid or base addition salts thereof, the
stereochemically isomeric forms thereof and the N-oxide form thereof, wherein :
X is CH2, N-R7, S or O ;
R7 is selected from the group of hydrogen, alkyl, phenyl, phenylalkyl,
alkylcarbonyl, alkyloxycarbonyl and mono- and dialkylaminocarbonyl,
the phenyl and alkyl groups being optionally substituted with one or more
halo atoms;
R1 and R2 are each, independently from each other, selected from the group of
hydrogen, hydroxy, cyano, halo, OSO2H, OS02CH3, phenyl, phenylalkyl,
alkyloxy, alkyloxyalkyloxy, alkyloxyalkyloxyalkyloxy,
tetrahydrofuranyloxy, alkylcarbonyloxy, alkylthio,
alkyloxyalkylcarbonyloxy, pyridinylcarbonyloxy,
alkylcarbonyloxyalkyloxy, alkyloxycarbonyloxy, alkenyloxy, alkenylcarbonyloxy
and mono-and dialkylaminoalkyloxy, the alkyl and aryl
radicals being optionally substituted with one or more hydroxy or halo
atoms or amino groups ; or
R1 and R2 may be taken together to form a bivalent radical -R'-R2- selected from the
group of -CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-O-, -CH2-O-CH2- and
-0-CH2-CH2-O- ;
a and b are asymmetric centers ;
(CH2)m is a straight hydrocarbon chain of m carbon atoms, m being an integer
ranging from 1 to 4 ;
Pir is an optionally substituted radical according to any one of Formula (Ha)wherein :
each R8 is independently from each other, selected from the group of
hydrogen, hydroxy, amino, nitro, cyano, halo and alkyl;
n is an integer ranging from 1 to 5 ;
R9 is selected from the group of hydrogen, alkyl and formyl; and
R3 represents an optionally substituted aromatic homocyclic or heterocyclic
ring system together with an optionally substituted and partially or
completely hydrogenated hydrocarbon chain of 1 to 6 atoms long with
which said ring system is attached to the Pir radical and of which may
contain one or more heteroatoms selected from the group of O, N and S.
More in particular, the invention relates to compounds according to Formula (I)
wherein R3 is a radical according to any one of Formula (Ilia), (Hlb) and (Hie)
(Figure Removed)
wherein :
d is a single bond while Z is a bivalent radical selected from the group of
-CHr, -C(=0)-, -CH(OH)-, -C(=N-OH)-, -CH(alkyl)-, -O-, -S-, -S(=O),
-NH- and -SH-; or d is a double bond while Z is a trivalent radical of
formula =CH- or =C(alkyl)-;
A is a 5- or 6-membered aromatic homocyciic or heterocyclic ring, selected
from the group of phenyl, pyranyl, pyridinyl, pyrazinyl, pyrimidinyl,
pyridazinyl, thienyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl,
oxadiazolyl and isoxazolyl;
p is an integer ranging from 0 to 4 ;
q is an integer ranging from 0 to 7 ;
R4 is selected from the group of hydrogen, alkyl, phenyl, biphenyl, naphthyl,
halo and cyano, the alkyl and aryl radicals being optionally substituted with
one or more hydroxy or halo atoms or amino groups ;
R5 is equal to R4 ; or
R4 and R5 may be taken together to form a bivalent radical -R4-R5- selected from the
group of -CH2-, =CH-, -CH2-CH2-, -CH=CH-, -O-, -NH-, =N-, -S-,
-CH2N(-alkyl)-, -CH=N-, -CH2O- and -OCH2-;
each R6 is independently from each other, selected from the group of hydrogen,
hydroxy, amino, nitro, cyano, halo, carboxyl, alkyl, phenyl, alkyloxy,
phenyloxy, alkylcarbonyloxy, alkyloxycarbonyl, alkylthio, mono- and
dialkylamino, alkylcarbonylamino, mono- and dialkylaminocarbonyl,
mono- and dialkylaminocarbonyloxy, mono- and dialkylaminoalkyloxy, the
alkyl and aryl radicals being optionally substituted with one or more
hydroxy or halo atoms or amino groups ; or
two vicinal radicals R6 may be taken together to form a bivalent radical -R6-R6-
selected from the group of-CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-C(=O)-,
-0-CH2-O-, -CH2-0-CH2-, -0-CH2-CH2-O-, -CH=CH-CH=CH-,
-CH=CH-CH=N-, -CH=CH-N=CH-, -CH=N-CH=CH-, -N=CH-CH=CH-,
-CH2-CH2-CH2-, -CH2-CH2-C(=O)- and -CH2-CH2-CH2-CH2-; and
R10 is selected from the group of hydrogen, alkyl, phenylalkyl and phenyl.
Preferably, the invention relates to those compounds wherein X=O or NH ; R1 and R2
are both alkyloxy ; m = 1; Pir is a radical according to Formula (Ila) wherein R8 is
hydrogen and n = 4 ; R3 is a radical according to Formula (mb) wherein Z is =CH-, d is
a double bond, A is a phenyl ring, R4 is an alkyl and R1 is hydrogen.
More preferably, the invention relates to compounds where X=O, R1 and R2 are both
methoxy ; m = 1; Pir is a radical according to Formula (Ha) wherein R8 is hydrogen
and n = 4 ; R3 is a radical according to Formula (ITlb) wherein Z is =CH-, d is a double
bond, A is a phenyl ring, R4 is methyl and R10 is hydrogen
In the framework of this application, alkyl defines straight or branched saturated
hydrocarbon radicals having from 1 to 6 carbon atoms, for example methyl, ethyl,
propyl, butyl, 1-methylpropyl, 1,1-dimethylethyl, pentyl, hexyl; or alkyl defines cyclic
saturated hydrocarbon radicals having from 3 to 6 carbon atoms, for example
cyclopropyl, methylcyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Halo is
generic to fluoro, chloro, bromo and iodo. Alkyl radicals being optionally substituted
with one or more halo atoms are for example polyhaloalkyl radicals, for example
difluoromethyl and trifluoromethyl.
The pharmaceutically acceptable salts are defined to comprise the therapeutically active
non-toxic acid addition salts forms that the compounds according to Formula (I) are
able to form. Said salts can be obtained by treating the base form of the compounds
according to Formula (I) with appropriate acids, for example inorganic acids, for
example hydrohalic acid, in particular hydrochloric acid, hydrobromic acid, sulfuric
acid, nitric acid and phosphoric acid ; organic acids, for example acetic acid,
hydroxyacetic acid, propanoic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid,
succinic acid, maleic acid, fumaric acid, malic acid, tartaric acid, citric acid,
methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic
acid, cyclamic acid, salicyclic acid, p-aminosalicylic acid and pamoic acid.
The compounds according to Formula (I) containing acidic protons may also be
converted into their therapeutically active non-toxic metal or amine addition salts forms
by treatment with appropriate organic and inorganic bases. Appropriate base salts
forms comprise, for example, the ammonium salts, the alkaline and earth alkaline metal
salts, in particular lithium, sodium, potassium, magnesium and calcium salts, salts with
organic bases, e.g. the benzathine, N-methyl-D-glucamine, hybramine salts, and salts
with amino acids, for example arginine and lysine.
Conversely, said salts forms can be converted into the free forms by treatment with an
appropriate base or acid.
The term addition salt as used in the framework of this application also comprises the
solvates that the compounds according to Formula (I) as well as the salts thereof, are
able to form. Such solvates are, for example, hydrates and alcoholates.
The N-oxide forms of the compounds according to Formula (I) are meant to comprise
those compounds of Formula (I) wherein one or several nitrogen atoms are oxidized to
the so-called N-oxide, particularly those N-oxides wherein one or more nitrogens of the
piperazinyl radical are N-oxidized.
The term "stereochemically isomeric forms" as used hereinbefore defines all the
possible isomeric forms that the compounds of Formula (I) may possess. Unless
otherwise mentioned or indicated, the chemical designation of compounds denotes the
mixture of all possible stereochemically isomeric forms, said mixtures containing all
diastereomers and enantiomers of the basic molecular structure. More in particular,
stereogenic centers may have the R- or S-configuration; substituents on bivalent cyclic
(partially) saturated radicals may have either the cis- or trans-configuration.
Compounds encompassing double bonds can have an E or Z-stereochemistry at said
double bond. Stereochemically isomeric forms of the compounds of Formula (I) are
obviously intended to be embraced within the scope of this invention.
Following CAS nomenclature conventions, when two stereogenic centers of known
absolute configuration are present in a molecule, an R or S descriptor is assigned (based
on Cahn-Ingold-Prelog sequence rule) to the lowest-numbered chiral center, the
reference center. The configuration of the second stereogenic center is indicated using
relative descriptors [R*,R* J or [R*,S*], where R* is always specified as the reference
center and [R*S*] indicates centers with the same chirality and [R*,S*] indicates
centers of unlike chirality. For example, if the lowest-numbered chiral center in the
molecule has an 5 configuration and the second center is R, the stereo descriptor would
be specified as S-[R*,S*]. If "a" and "(3" are used : the position of the highest priority
substituent on the asymmetric carbon atom in the ring system having the lowest ring
number, is arbitrarily always in the "a" position of the mean plane determined by the
ring system. The position of the highest priority substituent on the other asymmetric
carbon atom in the ring system (hydrogen atom in compounds of Formula® ) relative
to the position of the highest priority substituent on the reference atom is denominated
"a", if it is on the same side of the mean plane determined by the ring system, or "B", if
it is on the other side of the mean plane determined by the ring system.
Compounds of Formula (I) and some of the intermediates have at least two stereogenic
centers in their structure, respectively denoted a and b in Formula (I). Due to the
synthetic pathway followed for the synthesis of the tricyclic system, the configuration
of those two asymmetric centers a and b is predetermined, so that the relative
configuration of center a is S* and of center b is R*.
The invention also comprises derivative compounds (usually called "pro-drugs") of the
pharmacologically-active compounds according to the invention, which are degraded in
vivo to yield the compounds according to the invention. Pro-drugs are usually (but not
always) of lower potency at the target receptor than the compounds to which they are
degraded. Pro-drugs are particularly useful when the desired compound has chemical
or physical properties that make its administration difficult or inefficient. For example,
the desired compound may be only poorly soluble, it may be poorly transported across
the mucosal epithelium, or it may have an undesirably short plasma half-life. Further
discussion on pro-drugs may be found in Stella, V. J. et al., "Prodrugs", Drug Delivery
Systems, 1985, pp. 112-176, and Drugs, 1985,29, pp. 455-473.
Pro-drugs forms of the pharmacologically-active compounds according to the invention
will generally be compounds according to Formula (I), the pharmaceutically acceptable
acid or base addition salts thereof, the stereochemically isomeric forms thereof and the
N-oxide form thereof, having an acid group which is esterified or amidated. Included
in such esterified acid groups are groups of the formula -COOR*, where Rx is a
Chalky], phenyl, benzyl or one of the following groups :
Amidated groups include groups of the formula - CONRyRz, wherein Ry is H,
l, phenyl or benzyl and Rz is -OH, H, Chalky!, phenyl or benzyl.
Compounds according to the invention having an amino group may be derivatised with
a ketone or an aldehyde such as formaldehyde to form a Mannich base. This base will
hydrolyze with first order kinetics in aqueous solution.
The compounds of Formula (I) as prepared in the processes described below may be
synthesized in the form of racemic mixtures of enantiomers that can be separated from
one another following art-known resolution procedures. The racemic compounds of
Formula (I) may be converted into the corresponding diastereomeric salt forms by
reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently
separated, for example, by selective or fractional crystallization and the enantiomers are
liberated therefrom by alkali. An alternative manner of separating the enantiomeric
forms of the compounds of Formula (I) involves liquid chromatography using a chiral
stationary phase. Said pure stereochemically isomeric forms may also be derived from
the corresponding pure stereochemically isomeric forms of the appropriate starting
materials, provided that the reaction occurs stereospecifically. Preferably if a specific
stereoisomer is desired, said compound would be synthesized by stereospecific
methods of preparation. These methods will advantageously employ enantiomerically
pure starting materials.
The compounds according to the invention have surprisingly been shown to have
selective serotonine (5-HT) reuptake inhibitor activity in combination with additional
cc2-adrenoceptor antagonist activity and show a strong anti-depressant and/or anxiolytic
activity and/or a body weight control activity without being sedative. Also, in view of
their selective serotonine (5-HT) reuptake inhibitor as well as a2-adrenoceptor
antagonist activity, compounds according to the invention are also suitable for
treatment and/or prophylaxis in diseases where either one of the activities alone or the
combination of said activities may be of therapeutic use. In particular, the compounds
according to the invention may be suitable for treatment and/or prophylaxis in the
following diseases :
• Central nervous system disorders, including :
• Mood disorders, including particularly major depressive disorder, depression
with or without psychotic features, catatonic features, melancholic features,
atypical features of postpartum onset and, in the case of recurrent episodes, with
or without seasonal pattern, dysthymic disorder, bipolar I disorder, bipolar n
disorder, cyclothymic disorder, recurrent brief depressive disorder, mixed
affective disorder, bipolar disorder not otherwise specified, mood disorder due
to a general medical condition, substance-induced mood disorder, mood
disorder not otherwise specified, seasonal affective disorder and premenstrual
dysphoric disorders.
• Anxiety disorders, including panic attack, agoraphobia, panic disorder without
agoraphobia, agoraphobia without history of panic disorder, specific phobia,
social phobia, obsessive-compulsive disorder, posttraumatic stress disorder,
acute stress disorder, generalized anxiety disorder, anxiety disorder due to a
general medical condition, substance-induced anxiety disorder and anxiety
disorder not otherwise specified.
Stress-related disorders associated with depression and/or anxiety, including
acute stress reaction, adjustment disorders (brief depressive reaction, prolonged
depressive reaction, mixed anxiety and depressive reaction, adjustment disorder
with predominant disturbance of other emotions, adjustment disorder with
predominant disturbance of conduct, adjustment disorder with mixed
disturbance of emotions and conduct, adjustment disorders with other specified
predominant symptoms) and other reactions to severe stress.
Dementia, amnesic disorders and cognitive disorders not otherwise specified,
especially dementia caused by degenerative disorders, lesions, trauma,
infections, vascular disorders, toxins, anoxia, vitamin deficiency or endocrinic
disorders, or amnesic disorders caused by alcohol or other causes of thiamin
deficiency, bilateral temporal lobe damage due to Herpes simplex encephalitis
and other limbic encephalitis, neuronal loss secondary to anoxia/ hypoglycemia
severe convulsions and surgery, degenerative disorders, vascular disorders or
pathology around ventricle HI.
Cognitive disorders due to cognitive impairment resulting from other medical
conditions.
Personality disorders, including paranoid personality disorder, schizoid
personality disorder, schizotypical personality disorder, antisocial personality
disorder, borderline personality disorder, histrionic personality disorder,
narcissistic personality disorder, avoidant personality disorder, dependent
personality disorder, obsessive-compulsive personality disorder and personality
disorder not otherwise specified.
Schizoaffective disorders resulting from various causes, including
schizoaffective disorders of the manic type, of the depressive type, of mixed
type, paranoid, disorganized, catatonic, undifferentiated and residual
schizophrenia, schizophreniform disorder, schizoaffective disorder, delusional
disorder, brief psychotic disorder, shared psychotic disorder, substance-induced
psychotic disorder and psychotic disorder not otherwise specified.
Akinesia, akinetic-rigid syndromes, dyskinesia and medication-induced
parkinsonism, Gilles de la Tourette syndrome and its symptoms, tremor, chorea,
myoclonus, tics and dystonia.
Attention-deficit / hyperactivity disorder (ADHD).
Parkinson's disease, drug-induced Parkinsonism, post-encephalitic
Parkinsonism, progressive supranuclear palsy, multiple system atrophy,
corticobasal degeneration, parkinsonism-ALS dementia complex and basal
ganglia calcification.
Dementia of the Alzheimer's type, with early or late onset, with depressed
mood.
• Behavioral disturbances and conduct disorders in dementia and the mentally
retarded, including restlessness and agitation.
• Extra-pyramidal movement disorders.
• Down's syndrome.
• Akathisia.
• Eating Disorders, including anorexia nervosa, atypical anorexia nervosa,
bulimia nervosa, atypical bulimia nervosa, overeating associated with other
psychological disturbances, vomiting associated with other psychological
disturbances and non-specified eating disorders.
• AIDS-associated dementia.
Chronic pain conditions, including neuropathic pain, inflammatory pain, cancer
pain and post-operative pain following surgery, including dental surgery. These
indications might also include acute pain, skeletal muscle pain, low back pain,
upper extremity pain, fibromyalgia and myofascial pain syndromes, orofascial pain,
abdominal pain, phantom pain; tic douloureux and atypical face pain, nerve root
damage and arachnoiditis, geriatric pain, central pain and inflammatory pain.
Neurodegenerative diseases, including Alzheimer's disease, Huntington's chorea,
Creutzfeld-Jacob disease, Pick's disease, demyelinating disorders, such as multiple
sclerosis and ALS, other neuropathies and neuralgia, multiple sclerosis,
amyotropical lateral sclerosis, stroke and head trauma.
Addiction disorders, including:
• Substance dependence or abuse with or without physiological dependence,
particularly where the substance is alcohol, amphetamines, amphetamine-like
substances, caffeine, cannabis, cocaine, hallucinogens, inhalants, nicotine,
opioids, phencyclidine, phencyclidine-like compounds, sedative-hypnotics,
benzodiazepines and/or other substances, particularly useful for treating
withdrawal from the above substances and alcohol withdrawal delirium.
• Mood disorders induced particularly by alcohol, amphetamines, caffeine,
cannabis, cocaine, hallucinogens, inhalants, nicotine, opioids, phencyclidine,
sedatives, hypnotics, anxiolitics and other substances.
• Anxiety disorders induced particularly by alcohol, amphetamines, caffeine,
cannabis, cocaine, hallucinogens, inhalants, nicotine, opioids, phencyclidine,
sedatives, hypnotics, anxiolitics and other substances and adjustment disorders
with anxiety.
• Smoking cessation.
• Body weight control, including obesity.
• Sleep disorders and disturbances, including :
• Dyssomnias and/or parasomnias as primary sleep disorders, sleep disorders
related to another mental disorder, sleep disorder due to a general medical
condition and substance-induced sleep disorder.
• Circadian rhythms disorders.
• Improving the quality of sleep.
• Sexual dysfunction, including sexual desire disorders, sexual arousal disorders,
orgasmic disorders, sexual pain disorders, sexual dysfunction due to a general
medical condition, substance-induced sexual dysfunction and sexual dysfunction
not otherwise specified.
The present invention thus also relates to compounds of Formula (I) as defined
hereinabove, the pharmaceutically acceptable acid or base addition salts thereof, the
stereochemically isomeric forms thereof, the N-oxide form thereof, as well as the prodrugs
thereof for use as a medicine. Further, the present invention also relates to the
use of a compound of Formula (I), the pharmaceutically acceptable acid or base
addition salts thereof, the stereochemically isomeric forms thereof, the N-oxide form
thereof as well as the pro-drugs thereof for the manufacture of a medicament for
treating depression, anxiety and body weight disorders or more generally any one of the
diseases mentioned above.
The compounds according to the invention may also be suitable as add-on treatment
and/or prophylaxis in the above listed diseases in combination with antidepressants,
anxiolytics and/or antipsychotics which are currently available or in development or
which will become available in the future, to improve efficacy and/or onset of action.
This is evaluated in rodent models in which antidepressants, anxiolytics and/or
antipsychotics are shown to be active. For example, compounds are evaluated in
combination with antidepressants, anxiolytics and/or antipsychotics for attenuation of
stress-induced hyperthermia.
The invention therefore also relates to a pharmaceutical composition comprising the
compounds according to the invention and one or more other compounds selected from
the group of antidepressants, anxiolytics and antipsychotics as well as to the use of such
a composition for the manufacture of a medicament to improve efficacy and/or onset of
action in the treatment of depression and/or anxiety.
In vitro receptor and neurotransmitter transporter binding and signal-transduction
studies can be used to evaluate the a2-adrenoceptor antagonism activity and serotonine
(5-HT) reuptake inhibitor activity of the present compounds. As indices for central
penetration and potency to block the a2-adrenoceptors and serotonin transporters,
respectively, ex vivo a2-adrenoceptor and serotonin transporter occupancy can be used.
As indices of (X2-adrenoceptor antagonism in vivo, the reversal of the loss of righting
reflex, observed in rats after subcutaneous injection or oral dosage of the compound
before intravenous medetomidine administration in rats can be used (medetomidinetest).
As indices of serotonine (5-HT) reuptake inhibition activity, the inhibition of
head-twitches and excitation in rats, observed after subcutaneous injection or oral
dosage of the compound before subcutaneous p-chloroamphetamine administration in
rats can be used (pCA-test).
The invention also relates to a composition comprising a pharmaceutically acceptable
carrier and, as active ingredient, a therapeutically effective amount of a compound
according to the invention. The compounds of the invention or any subgroup thereof
may be formulated into various pharmaceutical forms for administration purposes. As
appropriate compositions there may be cited all compositions usually employed for
systemically administering drugs. To prepare the pharmaceutical compositions of this
invention, an effective amount of the particular compound, optionally in addition salt
form, as the active ingredient is combined in intimate admixture with a
pharmaceutically acceptable carrier, which carrier may take a wide variety of forms
depending on the form of preparation desired for administration. These pharmaceutical
compositions are desirable in unitary dosage form suitable, in particular, for
administration orally, rectally, percutaneously, by parenteral injection or by inhalation.
For example, in preparing the compositions in oral dosage form, any of the usual
pharmaceutical media may be employed such as, for example, water, glycols, oils,
alcohols and the like in the case of oral liquid preparations such as suspensions, syrups,
elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin,
diluents, lubricants, binders, disintegrating agents and the like in the case of powders,
pills, capsules and tablets. Because of their ease in administration, tablets and capsules
represent the most advantageous oral dosage unit forms in which case solid
pharmaceutical carriers are obviously employed. For parenteral compositions, the
carrier will usually comprise sterile water, at least in large part, though other
ingredients, for example, to aid solubility, may be included. Injectable solutions, for
example, may be prepared in which the carrier comprises saline solution, glucose
solution or a mixture of saline and glucose solution. Injectable suspensions may also
be prepared in which case appropriate liquid carriers, suspending agents and the like
may be employed. Also included are solid form preparations that are intended to be
converted, shortly before use, to liquid form preparations. In the compositions suitable
for percutaneous administration, the carrier optionally comprises a penetration
enhancing agent and/or a suitable wetting agent, optionally combined with suitable
additives of any nature in minor proportions, which additives do not introduce a
significant deleterious effect on the skin. Said additives may facilitate the
administration to the skin and/or may be helpful for preparing the desired compositions.
These compositions may be administered in various ways, e.g., as a transdermal patch,
as a spot-on, as an ointment.
It is especially advantageous to formulate the aforementioned pharmaceutical
compositions in unit dosage form for ease of administration and uniformity of dosage.
Unit dosage form as used herein refers to physically discrete units suitable as unitary
dosages, each unit containing a predetermined quantity of active ingredient calculated
to produce the desired therapeutic effect in association with the required
pharmaceutical carrier. Examples of such unit dosage forms are tablets (including
scored or coated tablets), capsules, pills, powder packets, wafers, suppositories,
injectable solutions or suspensions and the like, and segregated multiples thereof.
The compounds according to the invention can generally be prepared by a succession
of steps, each of which is known to the skilled person.
In particular, the compounds according to Formula (I) with a Pir-radical according to
Formula (Ila) can be prepared by a nucleophilic substitution reaction with a substituted
piperazine according to Formula (V) on an intermediate of Formula (IV). These
reactions may be carried out in a reaction inert solvent such as dioxane,
methylisobutylketone or N,N'-dimethylformamide, in the presence of a suitable base
such as potassium carbonate, sodium carbonate or triethylamine, or even without a
base, using in this latter case excess of reagent of Formula (V). Convenient reaction
temperatures range between 100°C and 150°C.
(Figure Removed)
In compound (IV), L represents any suitable reactive leaving group, in particular halo,
such as chloro, bromo or iodo or sulfonyloxy, such as methylsulphonyloxy or
4-methylbenzenesulfonyloxy.
The compounds according to Formula (I) with a Pir-radical according to Formula (Ha)
can also be prepared by a 2-step reaction scheme in which an intermediate of Formula
(IV) is first reacted with a substituted piperazine according to Formula (VII) after
which the R3-radical is introduced into the molecule. Reaction conditions are similar to
those described above for compounds of Formula (VI).
(Figure Removed)
In compound (IV), L represents any suitable reactive leaving group, in particular halo,
such as chloro, bromo or iodo or sulfonyloxy, such as methylsulphonyloxy or
4-methylbenzenesulfonyloxy.
In intermediate compound (VII), one of the nitrogen function may also be protected,
e.g. by a tert-butyloxycarbonyl-group.
(step 2)
(Figure Removed)
In compound (IX), L represents any suitable reactive leaving group, in particular halo,
such as chloro, bromo or iodo or sulfonyloxy, such as methylsulphonyloxy or
4-methylbenzenesulfonyloxy. Also R3"CHO may be used as compound (IX).
The compounds according to Formula (I) with a Pir-radical according to Formula (Ha)
can also be prepared by a 2-step reaction scheme in which an intermediate of Formula
(VET) is reacted with an acid according to Formula (X), followed by a subsequent
reduction of the carbonyl-function of intermediate (XI). Reactions of step 1 may be
carried out in a reaction inert solvent, such as chloroform, dichloromethane,
tetrahydrofuran, dimethylformamide or a mixture thereof, using any of methods known
to a person skilled in the art using condensation reagents such as
l,l'-carbonyldiimidazole, N,N'-dicyclohexylcarbodiimide or by previous
transformation of carboxylic acid of Formula (X) into its corresponding acid chloride.
Reactions shown in step 2 can be performed using a suitable reducing agent, such as
lithium-aluminum hydride or aluminum hydride, in a suitable solvent, for example
tetrahydrofuran. Generally, these reactions are run at a temperature ranging between
-20°C and room temperature.
(Figure Removed)
In intermediate compounds (XI) and (XII), the A-group represents an optionally
substituted aromatic homocyclic or heterocyclic ring system including a partially or
completely hydrogenated hydrocarbon chain of maximum 5 atoms long of which one
or more carbon atoms may be replaced by one or more atoms selected from the group
of oxygen, nitrogen and sulphur, with which the ring system is attached to the Pir
radical that has been defined above.
The substituents R1 and R2 may be changed or interconverted into each other by
methods well known in the art, such as demethylation, acylation, esterification,
amination and amidation.
The starting materials and some of the intermediates are compounds that are either
commercially available or may be prepared according to conventional reaction
procedures generally known in the art. For example, intermediates of Formula (IV) in
which X=O may be prepared according to the following reaction scheme (Scheme 1):
Scheme 1
In intermediate compound (XIV), L represents any suitable reactive leaving group, in
particular halo, such as chloro, bromo or iodo or sulfonyloxy, such as
methylsulphonyloxy or4-methylbenzenesulfonyloxy. Furthermore, Alk in intermediate
compound (XIV) represents any C1-6alkyl-group, in particular an ethyl-group and m is
defined as in Formula (I).
Intermediates according to Formula (IV) in which X=NH may also be prepared in an
equivalent manner according to above step 1, provided that the intermediate compound
(Xin) is replaced by its amine-analog (XVI), preferably with the amine group protected
with e.g. a COCFa- group. The alkylation step may be carried out in a reaction inert
solvent, for example, tetrahydrofuran or dimethylformamide, in the presence of a
strong base, such as sodium or potassium hydride, and the addition of a crown-ether,
such as 18-crown-6 or 15-crown-5. Convenient reaction temperatures range between
room temperature and 60°C.
Intermediates of Formula (XVII) are converted to oximes of Formula (XVIQ) using artknown
techniques, such as using hydroxylamine hydrochloride in the presence of
NaHCO3 or pyridine in a reaction inert solvent, for example ethanol. Intermediate
(XVIII) is oxidized to its nitril oxide and undergoes in situ an intramolecular
cycloaddition, yielding compound of Formula (XIX). This oxidation can be carried out
using a sodium hypochlorite solution in the presence of triethylamine in an inert solvent
such as dichloromethane at room temperature. Oxidation can also be performed using
Chloramine-T (N-chloro-4-methyl-benzenesulfonamide, sodium salt), stirring and
heating in a solvent such as refluxing ethanol. At this stage the two stereocenters a and
b of Formula (I) are formed.
(Figure Removed)
Preparation of a compound of Formula (XX) can be achieved using procedures known
in the art, for instance by reduction of the carbonyl compound of Formula (XIX) in the
presence of a suitable reducing agent, for example, sodiumborohydride in a suitable
solvent, such as water, an alcohol, tetrahydrofuran or a mixture thereof, generally at
room temperature.
(Figure Removed)
Intermediate of Formula (IV) can be prepared from intermediate of Formula (XX)
using standard techniques. Thus, reaction with methanesulfonyl chloride or
4-methyIbenzenesulfonyl chloride in the presence of a base, such as triethylamine, in a
reaction inert solvent, for example dichloromethane, at reaction temperatures ranging
between 0°C and room temperature, yields the corresponding sulfonyloxy derivative
intermediate (IV). The corresponding halo-derivative can also be prepared, e.g. treating
intermediate of Formula (XX) with triphenylphosphine, in the presence of
tetrachloromethane, in a reaction inert solvent, such as tetrahydrofuran, stirring and
refluxing the mixture.
(Figure Removed)
It is evident that in the foregoing and in the following reactions, the reaction products
may be isolated from the reaction medium and, if necessary, further purified according
to methodologies generally known in the art such as extraction, crystallization and
chromatography. It is further evident that reaction products that exist in more than one
enantiomeric form, may be isolated from their mixture by known techniques, in
particular preparative chromatography, such as preparative HPLC. Typically,
intermediate compounds (IV) and final compounds according to Formula (I) may be
separated into their enantiomeric forms.
Compounds according to the invention in which X=CHz may be prepared according to
the following reaction scheme (Scheme 2) in which an intermediary compound
according to Formula (V) is first N-alkylated with a dihaloderivative of Formula (XX)
using standard techniques, in the presence or absence of a base and in an inert reaction
solvent, such as chloroform, dichloromethane or 1,2-dichloroethane, and at reaction
temperatures ranging between room temperature and SOT, yielding an intermediate of
Formula (XXI). An aldehyde of Formula (XXEf) was reacted with rerr-butylamine in an
aprotic solvent such as toluene, stirring and heating at reflux temperature with removal
of water using a standard device, such as a Dean-Stark water separator, yielding an
imine of Formula (XXTV). C-alkylation of intermediary compound of Formula (XXIV)
with intermediate of Formula (XXI) can be achieved in the presence of an alkyl-lithium
derivative, such as n-butyllithium, under an inert atmosphere and in a dry inert solvent,
such as tetrahydrofuran, at low temperatures ranging between -78°C and 0°C, yielding
an intermediate of Formula (XXV). The intermediate compound of Formula (XXVI)
may be prepared by reaction of compound of Formula (XXV) with hydroxylamine, in
the presence of a base such as sodium bicarbonate, in a solvent such as a lower alkylalcohol
like ethanol, generally at room temperature. Finally, the oxidation of the oxime
derivative of Formula (XXVI) to its nitril oxide and subsequent in situ cycloaddition to
give compound of Formula (XXVII), may be achieved by similar standard techniques
such as those described above for intermediate of Formula (XVIII) to give compounds
of Formula (XIX).
(Figure Removed)
(step 5)
It is evident that the reaction steps disclosed above may be adapted to the specific
reaction products. The reaction steps disclosed may be performed in any way known
to the skilled person, including in solution or as solid phase reactions, the latter during
which the reaction products are bound to a resin material and are - in a final cleavage
step - released from the resin material. Examples of such embodiments and
adaptations have been disclosed by way of the Examples further in this application.
The compound 3,3a,4,5-tetrahydronaphto[l,2-c]isoxazole-3-acetic acid (Formula (IV)
wherein each of R1 and R2 are H, m=0, X=CH2 and L=COOH) and has been disclosed
in Synthetic Communications. 27(16), 2733-2742 (1997) as an intermediate for the
syntheses of anti-inflammatory, analgesic and antipyretic compounds and is excluded
from patent protection.
The following examples illustrate the present invention without being limited thereto.
Experimental part
The carbon ring numbering system for the compounds according to Formula (I) used in this
application is as follows :
Pir ]— R3
Of some compounds the absolute stereochemical configuration of the stereogenic
carbon atom(s) therein was not experimentally determined. In those cases the
stereochemically isomeric form which was first isolated is designated as "A" and the
second as "B", without further reference to the actual stereochemical configuration.
However, said "A" and "B" isomeric forms can be unambiguously characterized by a
person skilled in the art, using art-known methods such as, for example, X-ray
diffraction. The stereogenic centers a and b in Formula (I) have respectively the ring
numbers 3a and 3.
Hereinafter, "DMF" is defined as A-dimethylformarnide, "DIPE" is defined as
diisopropyl ether, and "THF" is defined as tetrahydrofurane.
A. Preparation of the intermediate compounds
Example Al.a
Preparation of intermediate 1
A solution of 4-bromo-2-butenoic acid methyl ester (0.1647 mol) in DMF (50 ml) was
added dropwise to a mixture of 2-hydroxy-4,5-dimethoxy-benzaldehyde (0.0823 mol)
and K2CO3 (0.1647 mol) in DMF (200 ml). The reaction mixture was stirred for 2
hours at room temperature, filtered and the filtrate was evaporated to dryness. The
residue was washed in a 10% aqueous NaOH solution, then extracted with QHkCl2. The
separated organic layer was dried (Na2SC>4), filtered, and the solvent was evaporated.
The residue was washed with diethyl ether, then dried. Yielding: 20 g of intermediate 1
(87%).
Example Al.b
Preparation of intermediate 2
Hydroxylamine (0.045 mol) was added to a solution of intermediate 1 (0.041 mol) in
ethano] (150 ml). Pyridine (57 ml) was added. The reaction mixture was stirred for 2
hours at room temperature, then poured out into water and acidified with concentrated
HC1. This mixture was extracted with CH2Ch. The separated organic layer was dried
(NaaSCU), filtered, and the solvent was evaporated. Yielding: 11.7 g (96%, crude
yield). A sample (2 g) was purified by high-performance liquid chromatography over
silica gel (eluent: CHaC2/CHsOH 95/5). The pure fractions were collected and the
solvent was evaporated. The residue was washed with diethyl ether, then dried.
Yielding: 0.9 g intermediate 2 .
Example Al.c
(Figure Removed)
Preparation of intermediate 3
NaOCl, 5% (130 ml) was added dropwise to a mixture of intermediate 2 (0.037 mol)
and EtsN (1 ml) in CH2Cla (220 ml). The reaction mixture was stirred for 4 hours at
room temperature, then washed with water, dried (NaaSC^), filtered, and the filtrate
was evaporated. The residue was purified by short open column chromatography over
silica gel (eluent: CH2Cl2/2-propanone 100/0 and 95/5). The desired fractions were
collected and the solvent was evaporated. Yielding: 5.8 g (54%, used in next
traction step without further purification). A sample (2 g) was recrystallised from
EtOAc. The precipitate was filtered off and dried. Yielding: 1.7 g of intermediate 3.
Example Al.d
Preparation of intermediate 4
NaBtLt (0.043 mol) was added portionwise to a solution of intermediate 3 (0.017 mol)
in THF (50 ml) and H2O (5 ml), stirred and cooled on an ice-bath. The resulting
reaction mixture was stirred for 2 hours at room temperature. 2-Propanone was added
while stirring for 30 min. The reaction mixture was washed with water and extracted
with CHzCh. The separated organic layer was washed with brine, dried
filtered and the solvent evaporated. The residue was purified by short open column
chromatography over silica gel (eluent: CH2C12/CH3OH 95/5) and by high-performance
liquid chromatography over silica gel (eluent: CH2C12/CH3OH 98/2). The pure fractions
were collected and the solvent was evaporated. A sample (1.8 g) was treated with
diethyl ether, then dried. Yielding: 1.2 g of intermediate 4 (59%).
Example Al.e
Preparation of intermediate 5
Et3N (0.016 mol) was added to a solution of intermediate 4 (prepared according to A3)
(0.0109 mol) in CH2C12 (60 ml). The mixture was cooled in an ice-bath.
Methanesulfonyl chloride (0.012 mol) was added and the resulting reaction mixture
was stirred for 30 min. Then, the mixture was washed with water, dried (Na2SCO4),
filtered and the solvent was evaporated. Yielding: 3.5 g of intermediate 5 (82%, used
in next reaction step without further purification).
Example Al.f
Preparation o f intermediate 6 I T T
Reaction under N2 atmosphere. BBr3 (0.04368 mol) was added dropwise to a stirred
solution of intermediate 5 (prepared according to Al.e) (0.00873 mol) in CH2C12 (100
ml), cooled to -78 °C. The reaction mixture was allowed to warm to
-40 °C and stirring was continued for 2 hours at -40 °C. Then, the mixture was poured
out into ice-water and extracted with CH2C12. The separated organic layer was dried
(MgSO4), filtered and the solvent evaporated. The residue was purified by flash column
chromatography over silica gel (eluent: CH2C12/CH3OH 97/3), then by HPLC (eluent:
CH2C12/CH3OH 99.5/0.5 to 90/10). Two product fraction groups were collected and
their solvent was evaporated. Yield: 0.750 g of intermediate 6 (26%).
Example Al.g
Preparation of intermediate 7
A mixture of intermediate 5 (prepared according Aid) (0.0422 mol) and piperazine
(0.1267 mol) in 1,4-dioxane (15 ml) was stirred for 2 hours at 100 °C. The solvent was
evaporated. The residue was washed with water and extracted with CH2Cb- The
separated organic layer was dried (NaaSO/t), filtered and the solvent was evaporated.
Yielding: 13 g of intermediate 7 (NMR: 85%).
Example Al.h
Preparation of intermediate 8
Intermediate 5 (prepared according to Al.e) (200 g, 0.58 mol) was separated into its
enantiomers by chiral column chromatography over column
LCI 10-2 with stationary phase CfflRALPAK-AD (2000 g, packing pressure: 45 bar,
detector range: 2.56, wavelength: 240nm, temperature: 30 °C; injection solution:
200 g in 8.4 L CH3CN; then, 19.6 L methanol (+ 2% ethanol) was added, then filtered;
injection-volume: 700 ml; eluent: CH3OH/CH3CN 70/30 v/v). Two product fraction
groups were collected and their solvent was evaporated. Yield: 95 g of intermediate 8 .
Example Al.i
Preparation of intermediate 9
A mixture of intermediate 8 (prepared according Al.h) (0.0728 mol) and l-(tertbutyloxycarbonyl)
piperazine (0.087 mol) in dioxane (500ml) was stirred and refluxed
for 48 hours. The solvent was evaporated and CH2Cla was added. HaO and NaOH
(50%) were added also and the mixture was extracted with CH2C12. The separated
organic layer was dried (MgSCU) and the solvent was evaporated in vacuum. Yield
intermediate 9
Example Al.i
Preparation of intermediate 10
A mixture of intermediate 9 (0.00318 mol) and 2,2,2-trifluoroacetic acid (189ml) in
CH2Ck (500ml) was stirred for 1 hour at room temperature. The solvent was
evaporated and the residue was dissolved in CH2Ck- NaOH (50%)was added and the
mixture was extracted. The separated organic layer was dried(MgSO4), filtered and the
solvent was evaporated in vacuum. The residue was purified by short column
chromatography over silica gel (eluent: CH2CJ2/(MeOH/NH3) 100/0;95/5). The pure
fractions were collected and the solvent was evaporated. Yield : 14.32g of intermediate
10 (59%).
Example Al.k
Preparation of intermediate 11
A mixture of intermediate 10 (0.00599 mol), l-Chloro-2-propanone (0.00599 mol) and
K2CO3 (0.01199 mol) in DMF (200 ml) was stirred for 24 hours at room temperature.
The solvent was evaporated. The residue was dissolved in CH2C12. The organic
solution was washed with water, dried (MgSCU), filtered and the solvent was
evaporated in vacuo. Yield: (quantitative yield) of intermediate 11 .
Example A 1.1
Preparation of intermediate 12
Reaction under N2 atmosphere. A mixture of 3,4-Dihydro-2-naphthalenecarboxylic
acid (0.0043 mol) and l,l'-carbonylbis[lH-imidazole] (0.0047 mol) in CH2C12) dry
was stirred for one hour at room temperature. A solution of intermediate 7 (prepared
according to Al.g) (0.0043 mol) in CH2C12, dry was added and the resulting reaction
solution was stirred for ± 24 hours at room temperature. The solution was washed with
water, then extracted with CH2C12. The separated organic layer was dried (Na2SO4),
filtered and the solvent was evaporated. The residue was purified by short open column
chromatography over silica gel (eluent: CH2C12/CH3OH 97/3). The pure fractions were
collected and the solvent was evaporated. Yield: 0.4 g of intermediate 12 (22%).
Example Al.m
Preparation of intermediate 13
Ethenyltriphenylphosphonium bromide (0.0025 mol) was added to a solution of
intermediate 7 (prepared according Al.g) (0.003 mol) in CH2C12 (20 ml). The reaction
mixture was stirred for 4 hours at room temperature. The solvent was evaporated under
reduced pressure. Yield: 2.2 g of intermediate 13 , used in next reaction step, without
further purification.
Example Al.n
Preparation of intermediate 14
To a solution of (E)-3-iodo-2-methylpropenoic acid (0.009 mol) in CH2Ch, dry
(100ml) at room temperature underNj flow, l.l'-carbonylbisflH-imidazole]
(0.0099 mol) was added. The mixture was stirred for 1 hour, then intermediate 7
(prepared according to Al.g) (0.009 mol) was added. The reaction mixture was stirred
at room temperature for 16 hours, washed with H2O and brine, dried (Na2SCO4) and the
solvent was evaporated under reduced pressure. The residue (white foam) was purified
by short open column chromatography over silica gel (eluent: CPfcCh/MeOH 99/1).
The pure fractions were collected and the solvent was evaporated. Yield : 3.82g of
intermediate 14 (white solid, 81%).
Example Al.o
Preparation of intermediate 15
A solution of LiAlHU, 1.0 M/THF (0.00848 mol) in THF (100ml) was stirred and
reffuxed under N2 flow at -20°C. A1C13 (0.0093 mol) was added in one portion and the
resulting mixture was stirred at -20°C for 10 minutes. A solution of intermediate 14
(prepared according to Al.n) (0.0077 mol) in THF (100ml) was added dropwise and
the resulting mixture was stirred at -20°C for 1 hour. A saturated NlitCl-solution 20%
was added dropwise at -10°C and the reaction mixture was allowed to warm to room
temperature. H2O was added to the suspension and was extracted with CH2Ch. The
separated organic layer was dried (Na2SCO4) and the solvent was evaporated under
reduced pressure. The residue was treated with Et2O and dried. Yield : 3.73g of
intermediate 15 (white solid, 94%).
Example Al.p
Preparation of intermediate 16
A mixture of intermediate 7 (prepared according Al.g) (0.015 mol), l-chloro-2-
propanone (0.015 mol) and K2CO3 (0.030 mol) in CH3CN (60 ml) was stirred for 24
hours at room temperature. The solvent was evaporated. The residue was partitioned
between water and CHaCh. The separated organic layer was dried (NaaSOt), filtered
and the solvent evaporated. The residue was purified by short open column
chromatography over silica gel (eluent: CHaCb/CEbOH 95/5). The pure fractions were
collected and the solvent was evaporated. Yield: 4.79 g of intermediate 16 (82%).
Example Al.q
Preparation of intermediate 17
.0128 mol) was added portionwise to a solution of intermediate 16 (prepared
according Al.p) (0.0051 mol) and H2O (3.2 ml) in THF (40.5 ml), at 0 °C. The reaction
mixture was stirred overnight at room temperature, then treated with a 10% aqueous
NHtCl solution. This mixture was extracted with CB2C2. The separated organic layer
was dried, filtered and the solvent evaporated. The residue was purified by short open
column chromatography over silica gel (eluent: CKbCh/CH/jOH 97/3). The desired
fractions were collected and the solvent was evaporated. Yield: 1.6 g of intermediate
17 (80%).
Example Al.r
Preparation of intermediate 18
Intermediate 7 (prepared according Al.g) (0.03 mol) was dissolved in CH3CN (200
ml) and K2COa (0.27 ml) was added. Oxiranemethanol (0.27 mol) was added and the
reaction mixture was stirred over the weekend at 60°C. The solvent was evaporated in
vacuo. The residue was partitioned between water and CH2C12. The organic layer was
separated, dried (Na2SO4), filtered and the solvent was evaporated under reduced
pressure. The residue was purified by preparatory HPLC ((1) eluent:
CH2C12/(CH3OH/NH3) 95/5, then (2) eluent: CH2C12/CH3OH 90/10). The product
fractions were collected and the solvent was evaporated. Yield: 7.5 g (61%) of pure
intermediate 18 and 3.5 g of a mixture of starting material/target compound 1/1.
Example Al.s
Preparation of intermediate 19
Intermediate 18 (prepared according Al.r) (0.0012 mol) was dissolved in CH2Cl2 (20
ml). A solution of periodic acid sodium salt (0.0024 mol) in NaHCO3/H20 (q.s.) was
added and the resulting reaction mixture was stirred vigorously for 2 hours. The
mixture was partitioned between water and CH2O2. The separated organic layer was
washed with brine, dried (Na2SCO4), filtered and the solvent evaporated in vacuo. Yield:
0.430 g of intermediate 19 (quantitative yield; used in next reaction step, without
further purification).
Example A2.a
Preparation of intermediate 20
Reaction under N2 atmosphere. A solution of 2,2,2-trifluoro-N-(2-formylphenyl)-
acetamide, (0.1869 mol) in DMF (375 ml) was added dropwise to NaH (0.2055 mol) in
DMF (375 ml). The mixture was stirred for 30 min. at room temperature. A solution of
4-Bromo-3-butenoic acid methyl ester (0.2803 mol) in DMF (200 ml) was added
dropwise. Then, 18-crown-6 (catalytic quantity) was added. The resulting reaction
mixture was stirred for 2 hours at 60 °C, then overnight at room temperature. The
solvent was evaporated. The residue was washed in water and extracted with CH2C12.
The separated organic layer was dried (Na2SO4), filtered and the solvent evaporated.
The residue was purified by open column chromatography over silica gel (eluent:
CH2Cl2/hexane 90/10, 100/0 and with CH2Cl2/2-propanone 96/4, 90/10 and 80/20). The
pure fractions were collected and the solvent was evaporated. Yielding: 44.37 g of
intermediate 20 (75%, used in next reaction step, without further purification).
Example A2.b
Preparation of intermediate 21
Hydroxylamine (0.169 mol) and pyridine (0.211 mol) were added to a solution of
intermediate 20 (prepared according to A2.a) (0.1407 mol) in ethanol (450 ml) and the
resulting reaction mixture was stirred for 3 hours at room temperature. The mixture was
washed with a 10% citric acid solution, then extracted with CH2Cl2. The separated
organic layer was dried (Na2SCO4), filtered and the solvent was evaporated. Yielding:
45.76 g of intermediate 21 (98%, used in next reaction step, without further
purification).
Example A2.c
Preparation of intermediate 22
or
F
A mixture of intermediate 21 (prepared according to A2.b) (0.0658 mol) and N-chloro-
4-methyl-benzenesulfonamide, sodium salt (0.0658 mol) in ethanol (500 ml) was
stirred and refluxed for 2 hours. The mixture was concentrated in vacuo, filtered over
dicalite, and the filtrate was washed with water and brine, then extracted with CH2C12.
The separated organic layer was dried (Na2SO), filtered and the solvent evaporated.
The residue was purified by open column chromatography over silica gel (eluent:
CH2Cl2/2-propanone 100/0, 96/4, 90/10 and 80/20). The desired fractions were
collected and the solvent was evaporated. The residue (syrup) was crystallized from
hexane, then washed with DIPE, and dried. Yielding: 12.32 g of intermediate 22
(57%).
Example A2.d
Preparation of intermediate 23
NaBHt (0.0289 mol) was added portionwise to a mixture of intermediate 22 (prepared
according to A2.c) (0.0116 mol) in THF (81 ml) and H2O (6.8 ml), stirred and cooled
on an ice-bath. The resulting reaction mixture was stirred overnight at room
temperature. The mixture was treated with a saturated aqueous NHtCl solution, then
extracted with EtOAc. The separated organic layer was dried (Na2SCO4), filtered and the
solvent was evaporated. The residue was washed with CH2C12, then recrystallized from
EtOAc. The precipitate was filtered off and dried. Yielding: 0.9 g of intermediate 23
(38%).
Example A2.e
Preparation of intermediate 24
A mixture of intermediate 23 (prepared according A2.d) (0.001468 mol) and
triphenylphosphine (0.001909 mol) in tetrachloromethane (30 ml) and THF (20 ml)
was stirred and refluxed for 3 hours. The solvent was evaporated till dryness. The
residue was purified by short open column chromatography over silica gel (eluent:
CH2Cl2/hexane 90/10, then 100/0). The desired fractions were collected and the solvent
was evaporated. The residue was crystallized from methanol. The precipitate was
filtered off and dried. Yielding: 2.6 g of intermediate 24 (79%).
Example A3.a
Preparation of intermediate 25 I V
1,1-Dimethylethyl 1-piperazinecarboxylate (0.02 mol) was added portionwise to a
solution of l,4-dichloro-2-butene (0.025 mol) in CHCls (60 ml). The reaction mixture
was stirred for 24 hours at room temperature, then stirred and refluxed for 24 hours.
The reaction was quenched with a saturated aqueous NaHCOs solution, dried (Na2SO4),
filtered and the solvent was evaporated. The residue was purified by short open column
chromatography over silica gel (eluent: CH2Cl2/EtOAc). The pure fractions were
collected and the solvent was evaporated. Yielding: 2.2 g of intermediate 25 (40%).
Example A3.b
Preparation of intermediate 26
Reaction was carried out under Nj flow. A mixture of NaH, 60% (0.0579 mol) and 18-
crown-6 (cat.quant.) in THF (25ml) was cooled. A mixture of 2-Amino-4,5-
dimethoxybenzaldehyde (0.0579 mol) in THF (50ml) was added portionwise. The
reaction was stirred at room temperature for 30 min. A mixture of intermediate 25
(prepared according to A3.a) (0.0386 mol) in THF (50ml) was added portionwise. The
mixture was stirred at room temperature for 3 days and then treated with NBUC1 (10%).
The mixture was extracted with CH2C12. The organic layer was separated, dried,
filtered and the solvent was evaporated till dryness. The residue was purified by short
open column chromatography (eluent: CH2C12/CH3OH 99/1 and 98/2). The pure
fractions were collected and the solvent was evaporated. Yield: 5.5g of intermediate 26
(23%).
Example A3.c
Preparation of intermediate 27
The reaction was carried out under N2 flow. A mixture of intermediate 26 (prepared
according A2.b) (0.02 mol) in THF (80ml) and 18-crown-6 (cat.quant.) were added
portionwise to a mixture of NaH, 60% (0.03 mol) in THF (20ml). The mixture was
stirred at room temperature for 20 min. trifluoroacetic acid anhydride (0.022 mol) was
added portionwise. The mixture was stirred at room temperature for 3 hours, treated
with a solution of NH4Cl (20%) and then extracted with CH2C12 and the solvent was
evaporated till dryness. The residue was purified by short column chromatography over
silica gel (CH2C12/CH3OH 99/1 and 98/2). The pure fractions were collected and the
solvent was evaporated. Yield: 5.3g of intermediate 27 (58%).
Example A3.d
Preparation of intermediate 28
A mixture of intermediate 27 (prepared according to A3.c) (0.0115 mol),
hydroxylamine (0.0126 mol) and NaHCO3 (0.023 mol) in ethanol, abs. (60ml) was
stirred at room temperature for 24 hours, filtered off and the solvent was evaporated till
dryness. Yield: 5.8g of intermediate 28 (95%).
Example A3.e
Preparation of intermediate 29
l-chloro-2,5-Pyrrolidinedione (0.0272 mol) was added portionwise to a solution of
intermediate 28 (prepared according to A3.d) (0.0109 mol) in CH2C12 (100ml). The
mixture was stirred at room temperature for 2 hours. Et3N (0.0272 mol) was added
dropwise. The mixture was stirred at room temperature overnight, quenched with a
K2CC3 10% solution, then extracted and the solvent was evaporated till dryness. Yield:
of intermediate 29 .
Example A3.f
Preparation of intermediate 30
A mixture of intermediate 29 (prepared according to A3.e) (0.0109 mol) and LiOH
(0.0119 mol) in H2O (17.5ml) and 1,4-dioxane (70ml) was stirred at room temperature
for 3 hours. The mixture was treated with a NaOH (2N) solution and then extracted
with CH2C12. The solvent was evaporated till dryness. The residue was purified by
short open column chromatography (eluent: CH2Cl2/CH3OH 98/2). The pure fractions
were collected and the solvent was evaporated. Yield: 2.07g of intermediate 30 (45%).
Example A3.g
N o
Preparation of intermediate 31
Trifluoroacetic acid (7.9ml) was added dropwise to a solution of intermediate 30
(0.0047 mol) in CH2C12 (33ml). The mixture was stirred at room temperature for 3
hours, cooled and basified with a 50% NaOH solution. The mixture was extracted and
the solvent was evaporated till dryness. Yield: 1.6g of compound 31 (100%).
Example A4.a
Preparation of intermediate 32
l,4-dichloro-2-butene (0.03 mol) was added to a mixture of l-(2-
naphthylmethyl)piperazine (0.025 mol) and NaHCO3 (0.025 mol) in CH2C12 (75 ml).
The reaction mixture was stirred for 24 hours at room temperature. The solid was
filtered off, washed with more CH2C12 and the organic solution was washed with a 10%
Na2CCO3 solution, dried (Na2SO4), filtered and the solvent was evaporated. The residue
was purified by short open column chromatography over silica gel (eluent:
CH2Cl2/EtOAc/2-propanone). The pure fractions were collected and the solvent was
evaporated. Yield: 3.4 g of intermediate 32 (43%).
Example A4.b
Preparation of intermediate 33
A solution of NS (0.0546 mol) and teit-Butylamine (0.0983 mol) in toluene (75 ml)
was stirred and refluxed for 24 hours using a Dean-Stark water separator. The solvent
was evaporated. The residue was purified biyi.S-tiUatjjSW (bp at 0.5 mm Hg: 75 °C).
Yielding: 8.1 g of intermediate 33 (72%).,
Example A4.c
Preparation of intermediate 34
Reaction under N2 atmosphere. n-BuLi (0.014 mol) was added dropwise to a solution
of intermediate 33 (prepared according A4.a) (0.0125 mol) and 2,2,6,6-
tetramethylpiperidine (0.0012 mol) in THF, dry (25 ml), stirred at -78 °C. The mixture
was stirred for 3 hours at -10 °C. A solution of intermediate 32 (prepared according
A4.b) (0.0083 mol) in THF, dry (25 ml) was added portionwise at -10 °C. The reaction
mixture was stirred for 24 hours at room temperature, then quenched with NHjCl
(10%) and the organic layer was separated. The aqueous layer was extracted with
CH2C12. The combined organic layers were dried (Na2SO4), filtered and the solvent was
evaporated. Yielding: 5.6 g of intermediate 34 (100%)
Example A4.d
Preparation of intermediate 35
NaHCO3 (0.015 mol) and hydroxylamine (0.0125 mol) were added to a solution of
intermediate 34 (prepared according A4.c) (0.0083 mol) in ethanol, abs. (50 ml). The
reaction mixture was stirred for 24 hours at room temperature. CHaC^ was added and
the solid was filtered off and washed with CH2Cl2. The solvent was evaporated. The
residue was taken up into QHCh and washed with 10% Na2CO3 and with brine. The
organic layer was separated, dried (Na2SCO4.), filtered and the solvent was evaporated.
The residue was purified by short open column chromatography over silica gel (eluent:
CH2Cl2/EtOAc/2-propanone). The desired fractions were collected and the solvent was
evaporated. Yielding: 0.9 g of intermediate 35 (24%).
Example A5.a
-N-^OO-b
Preparation of compound 36
A mixture of A (0.029 mol) and intermediates (prepared according to Al.e) (0.0058
mol) in 1,4-dioxane (5 ml) was stirred for 6 hours at 100 °C. The solvent was
evaporated. The residue was purified by short open column chromatography over silica
gel (eluent: CH2C12/CH3OH 98/2 - 97/3). The product fractions were collected and the
solvent was evaporated. Yield: 3.3 g of intermediate 36
Example A5.b
(Figure Removed)
Preparation of compound 37 ll if II
Trifluoroacetic acid (11.7 ml) was added dropwise to a solution of intermediate 36
(prepared according to A5.f) (0.0071 mol) in CHC13 (50 ml) and the resulting reaction
mixture was stirred for 3 hours at ± 10 °C. The reaction mixture was cooled, then
further alkalized with 50% NaOH. This mixture was extracted and the extract's solvent
was evaporated. Yield: 2.5 g of intermediate 37 (quantitative yield; used in next
reaction step, without further purification).
B. Preparation of the final compounds
Example Bl. a
Preparation of compound 1
A mixture of intermediates (prepared according to Al.e) (0.0291 mol) and l-(3-
phenyl-2-propenyl)-piperazine, (0.0582 mol) was heated for 2 hours at 100 °C. The
crude reaction mixture was washed with water and extracted with CH2C12. The
separated organic layer was dried (Na2SCO4), filtered and the solvent was evaporated.
The residue was purified by short open column chromatography over silica gel (eluent:
CH2C12/CH3OH 95/5) and by HPLC over silica gel (eluent: CH2C12/CH3OH 80/20).
The pure fractions were collected and the solvent was evaporated. This fraction was
separated into its optical enantiomers by chiral column chromatography over Chiralpak
AD (eluent: C2H5OH/CH3CN 90/10). The (B)-enantiomeric fractions were collected
and the solvent was evaporated. The residue was dissolved in methanol and converted
into the hydrochloric acid salt (1:2). The precipitate was filtered off and dried.
Yielding: 2.47 g of compound 1 .
ExampleBl.b
Preparation of compound 2
A mixture of intermediate 5 (prepared according to Al.e) (0.0044 mol) and (3-phenyl-
2-propenyl)-piperazine (0.0087 mol) was stirred for 2 hours at 100 °C. The reaction
mixture was purified by short open column chromatography over silica gel (eluent:
CH2C12/CH3OH 95/5), then by high-performance liquid chromatography over silica gel
(eluent: CH2Cl2/CH3OH 96/4). The pure fractions were collected and the solvent was
evaporated. The residue (1.4 g) was treated with diethyl ether, then dried. Yielding: 1.2
g of compound 2 (60%).
Example Bl.c
Preparation of compound 3
A mixture of intermediate 6 (0.00227 mol), (E) l-(2~methyl-3-phenyl-2-
propenyl)piperazine (0.00273 mol) and NaHCO3 (0.00455 mol) in dioxane (30 ml)
was stirred and refluxed for 48 hours. The solvent was evaporated. The residue was
dissolved in CH2C12. The organic solution was washed with water, dried (MgSCU),
filtered and the solvent was evaporated. The residue was purified by short open
column chromatography over silica gel (eluent: CH2C12/(CH3OH/NH3) 99/1), then by
HPLC (eluent: CH2C12/CH3OH 99.5/0.5 to 98/2). The desired fractions were collected
and the solvent was evaporated. Yield: 0.17 g of compound 3 .
Example Bl.d
Preparation of compound 4
B-CIS
A mixture of intermediate 8 (prepared according to Al.h) (0.0058 mol)
and HNx jt (0.0116 mol) in dioxane (10ml) was stirred and refluxed
for 8 hours, then stirred overnight at room temperature, then stirred and refluxed for 18
hours. The mixture was treated with H2O and extracted with CH2Cl2- The solvent of
the separated organic layer was evaporated. The residue was purified by short open
column chromatography (eluent: CHzClj/MeOH 97/3). The desired fractions were
collected and the solvent was evaporated. The residue was treated with diethyl ether,
then dried. Yield : 0.9g of compound 4 (33%).
Example Bl.e
Preparation of compound 5
A mixture of intermediate 10 (prepared according to Al.i) (0.0029 mol),
(0.0058 mol), AcOH (0.48g) and (AcO)3BHNa (0.4g)
in 1,2-dichloroethane (20ml) was stirred and refluxed overnight. The mixture was
treated with HaO and extracted. The solvent of the separated organic layer was
evaporated. The residue was purified by short column chromatography over silica gel
(eluent : CHaC12/MeOH 97/3). The desired fractions were collected and the solvent
was evaporated. The residue was treated with diethyl ether, then dried. Yield : l.OTg of
compounds (82%).
Example Bl.f
Preparation of compound 6
A mixture of compound 3 (prepared according ex. Bl) (0.00020 mol), acetylchloride
(0.00024 mol) and Et3N (0.00061 mol) in chloroform (10 ml) was stirred at room
temperature for 2 hours. Water was added and this mixture was extracted with CH2Cb-
The separated organic layer was dried (MgSCU), filtered and the solvent evaporated in
vacuo. The residue was purified by CC-TLC on Chromatotron (eluent:
CH2C12/CH3OH 97/3; then 99/1). The desired fractions were collected and the solvent
was evaporated. Yield: 0.022 g of compound 6 .
Example Bl.g
Preparation of compound 7
Preparation of compound 8
Compound 2 (prepared according Bl.b) (0.0022 mol) was separated and purified into
its optical enantiomers by chiral column chromatography over Chiralpak AD (eluent:
C2HsOH/CH3CN 90/10). Two fraction groups were collected and their solvent was
evaporated. Yielding: ± 1.5 g of fraction 1 (LCI purity: > 99.5%) and ± 1.5 g of
fraction 2 (LCI purity: > 99.5%). Fraction 1 was crystallized by treatment with hexane,
stirring overnight. The precipitate was filtered off and dried. Yielding: 1.08 g of
compound compound 7 (grease solid). Fraction 2 was crystallized by treatment with
EtOAc, stirring overnight. The precipitate was filtered off and dried. Yielding: 0.54 g
of 8 (grease solid).
Example B2.a
Preparation of compound 9
A mixture of intermediate 24 (prepared according A2.e) (0.0022 mol), l-(2-
naphthalenylmethyO-piperazine, (0.0044 mol) and KI (catalytic quantity) in 1,4-dioxane
(2.5 ml) was stirred and refluxed overnight. The reaction mixture was washed with
water and this mixture was extracted with CH2C The residue was purified by short
open column chromatography over silica gel (eluent: C2CWCHsOH 98/2), then by
HPLC (eluent: CH2Cl2/(CH3OH/NH3) 96/4). The pure fractions were collected and the
solvent was evaporated. The residue was treated with DIPE, filtered off and dried.
Yield: 0.3 g of compound 9 (30%).
Example B2.b
Preparation of compound 10
Reaction under N2 atmosphere. A solution of compound 9 (prepared according to
B2.a) (0.0012 mol) in THF, dry (3 ml) and 18-crown-6 (catalytic quantity) was slowly
added to a solution of NaH, 60% (0.0018 mol) in THF, dry (2 ml). The reaction
mixture was stirred for 30 min at room temperature, acetylchloride (0.0013 mol) was
added dropwise and the reaction mixture was stirred for 3 hours at room temperature.
The reaction mixture was treated with aqueous NH^Cl and extracted with CH2C12. The
separated organic layer was dried, filtered and the solvent evaporated. The residue was
purified by short open column chromatography over silica gel (eluent: CH2C12/CH3OH
98/2), then by HPLC (eluent: CH2C12/(CH3OH/NH3) 98/2). The pure fractions were
collected and the solvent was evaporated. Yield: 0.26 g of compound 10 (52%).
Example B3a
Preparation of compound 11
A mixture of intermediate 31 (prepared according A3.g) (0.0045 mol),
(E)- (3-chloro-2-methyl-l-propenyl)-benzene (0.0037 mol) and K2CO3 (0.0037 mol) in
DMF (15 ml) was stirred at 70°C for 2 hours. The mixture was washed with water and
then extracted with CH2Cl2 . The organic layer was separated, dried, filtered and the
solvent was evaporated. The residue was purified by short column chromatography
over silica gel (eluent: CH2C12/CH3OH 98/2). The desired fractions were collected and
the solvent was evaporated. The residue was purified again by HPLC (eluent:
CH2Cl2/(CH3OH/NH3) 98/2). The pure fractions were collected and the solvent was
evaporated. The residue was treated with DBPE. The precipitate was filtered off and
dried. Yield: 0.34 g of compound 11 (20%).
Example B3.b
Preparation of compound 12
Preparation of compound 13
Compound 11 (prepared according to B3.a) (0.00605 mol) was purified by highperformance
liquid chromatography over Chiralcel OJ (eluent: hexane/MeOH/EtOH
20/24/56). The desired fractions were collected and the solvent was evaporated. Yield
: fractions A and B. Fraction A was purified by high-performance liquid
chromatography over RP BBS C18 (eluent: (0.5%NH4OAc in
H2O/CH3CN(90/10))/MeOH 70/30). The pure fractions were collected and the organic
solvent was evaporated. The aqueous layer was extracted with CH2C12. The separated
organic layer was dried (MgSO4), filtered and the solvent was evaporated. The residue
was stirred in hexane and the precipitate was filtered off. Yield : 0.69g of compound
12 . Fraction B was purified by high-performance liquid chromatography over RP
BDS CIS (eluent: (0.5%NH4OAc in H2O/CH3CN(90/10))/MeOH 70/30). The pure
fractions were collected and the organic solvent was evaporated. The aqueous layer
was extracted with CH^Ch. The separated organic layer was dried (MgSCU), filtered
and the solvent was evaporated. The residue was stirred in hexane and the precipitate
was filtered off. Yield : 0.67g of compound 13 .
Example 4
Preparation of compound 14
Reaction done in solid phase using a Quest 210 synthesizer (Argonaut Technologies,
San Carlos, USA). N,N-Diisopropylethylamine (0.0036 mol) was added to a
suspension of
(°-0006 mo1)in acetonitrile (4 ml).
l-(2-Chlorophenylmethyl)piperazine (0.0012 mol) was added and the resulting reaction
mixture was stirred for 20 hours at 80 °C. Then, each reaction vessel was filtrated and
the filtrate was evaporated. The residue was HPLC purified. The pure fractions were
collected and the solvent was evaporated. Yield: 0.102 g of compound 14
Example B5.a
Preparation of compound 15
A mixture of intermediate 7 (prepared according Al.g) (0.0036 mol),
2-(bromomethyl)naphthalene (0.0055 mol) and K2CO3 (0.0055 mol) in MIK (15 ml)
was stirred for ± 24 hours at 100 °C. The crude reaction mixture was washed with
water, then extracted with CH2Cl2. The separated organic layer was dried (Na2SCO4),
filtered and the solvent was evaporated. The residue was purified by high-performance
liquid chromatography over silica gel (2 x) ((I) eluent: CH2C12/CH3OH 95/5; (II)
eluent: CH2C12/(CH3OH/NH3) 98/2). The pure fractions were collected and the solvent
was evaporated. Yielding: 0.2 g of compound 15 (11%).
ExampleBS.b
Preparation of compound 16
Preparation of compound 17
Compound 15 (prepared according B5.a) (0.0106 mol) was separated into its
enantiomers by column chromatography (eluent: hexaneAI^HsOH gradient 30/70 to
0/100; column: CfflRALPAK AD 1000 A 20 /Am DIACEL). Two pure fractions were
collected and their solvents were evaporated. The residue was dissolved in CH3OH and
converted into the hydrochloric acid salt (1:2). The precipitate was filtered off and
dried. Yielding: 2.08g of compound 16 (36%) and 2.19g of compound 17 (38%).
Example B6
Preparation of compound 18
A mixture of intermediate 7 (prepared' according to Al.g) (0.0045 mol),
2-methyl-3-(3-thienyl)-2-propenal (0.00675 mol), NaBH(OAc)3 (0.00675 mol) and
HO Ac (2 drops) in 1,2-dichloroethaan (30 ml) was stirred overnight at room
temperature. A saturated aqueous NHtCl solution was added and this mixture was
extracted with CHaCh. The separated organic layer was dried (MgSCO4), filtered and the
solvent evaporated in vacuo. The residue was purified by flash column chromatography
over silica gel (eluent: CH2C12/(CH3OH/NH3) 97/3), then by HPLC (eluent:
CH2Cl2/(CH3OH/NH3) 99/1 to 98/2). The product fractions were collected and the
solvent was evaporated. Yield: 0.965 g of compound 18 (46%; containing also 3% of
the (Z) isomer!).
Example B7
Preparation of compound 19
A mixture of intermediate 10 (prepared according to Al.j) (0.003 mol),
4-chlorobenzaldehyde (0.0045 mol) and (AcO)3BHNa (0.0045 mol) in
1,2-dichloroethane (30ml) was stirred and refluxed for 2 hours at room temperature. A
saturated aqueous NEUCl-solution was added and the mixture was extracted with
CH2C12. The separated organic layer was dried (MgSO4), filtered and the solvent was
evaporated in vacuum. The residue was purified by short open column chromatography
over silica gel (eluent: CH2Cl2/(MeOH/NH3) 97/3). The desired fractions were
collected and the solvent was evaporated. The residue was precipitated from DIPE.
Yield: 1.180g of compound 19 (57%).
Example B8
Preparation of compound 20
A mixture of (0.00036 mol),
2-methylbenzaldehyde (0.00108 mol) and NaOCH3, 30% in CH3OH (0.00108 mol) in
CH3OH, dry (8 ml) was stirred for 16 hours at 65 °C (Reaction done in solid phase
using a Quest 210 synthesizer (Argonaut Technologies, San Carlos, USA)). The
mixtures were filtered and the filtrate was HPLC purified (eluent:
CH2Cl2/(CH3OH/NH3) 98/2). The desired fractions were collected and the solvent was
evaporated. Yield: 0.032 g of compound 20.
Example B9
Preparation of compound 21
Reaction under N2 atmosphere. Solution LiAIKU, 1 M/THF (0.8 ml) was stirred at -20
°C. A1C13 (0.0009 mol) was added in one portion. The resulting solution was stirred for
10 min at -20 °C. A solution of intermediate 12 (prepared according to Al.l) (0.0008
mol) in THF, dry (5 ml) was added dropwise and the resulting reaction mixture was
stirred for one hour at -20 °C. Then; a saturated aqueous NH4Cl solution was added
carefully. The reaction mixture was washed with water, then extracted with CH2C12.
The separated organic layer was dried (Na2SCO4), filtered and the solvent was
evaporated. The residue was treated with ether. The residue (0.13 g) was purified by
CC-TLC Chromatotron (eluent: CH2C12/CH3OH 97/3). The pure fractions were
collected and the solvent was evaporated. Yield: 0.09 g of compound 21 (30%).
Example BIO
Preparation of compound 22
Reaction under N2 flow. A mixture of intermediate 13 (prepared according to Al.m)
(0.001 mol) in CH3OH, dry (20 ml) was stirred at room temperature. NaOCH3, 30% in
CH3OH (0.002 mol) was added. 5-Indanecarboxaldehyde (0.002 mol) was added and
the resulting reaction mixture was stirred and refluxed for 16 hours. The reaction
mixture was allowed to cool to room temperature. 20% NH4Cl was added and this
mixture was extracted with CH2Cl2. The separated organic layer was washed with
water, with brine, dried (NajSO4), filtered, and the solvent was evaporated under
reduced pressure. The residue was purified by short open column chromatography and
CC-TLC (eluent: CH2C12/CH3OH 98/2). The product fractions were collected and the
solvent was evaporated. Yield: 0.035 g of compound 22 (7.2%, light-brown solid).
Example B11
Preparation of compound 23
A mixture of intermediate 15 (prepared according to Al.o) (0.00136 mol), 2-
(trimethylstannyl)pyridine (0.0027 mol) and Pd(PPh3)4 (0.00013 mol) in toluene (20ml)
was heated to 100°C. The reaction mixture was stirred for 16 hours and was allowed to
cool to room temperature. H2O was added and the mixture was extracted with CH2C12.
The separated organic layer was collected, washed with H2O and brine, dried (Na2SO4)
and the solvent was evaporated under reduced pressure. The residue was purified by
short open column chromatography over silica gel (eluent: CH2Cl2/MeOH 98/2). The
pure fraction was collected and the solvent was evaporated. The resulting residue was
purified by CC-TLC on Chromatotron (eluent: CH2CL2/MeOH 98/2). The pure
fraction was collected and the solvent was evaporated. Yield : 0.044g of compound 23
(light brown solid, 7%).
ExampleB12
Preparation of compound 24
Reaction under N2 atmosphere. n-BuLi, 2.5M/hexanes (0.0062 mol) was added
dropwise to a stirred solution of (p-Flurorobenzyl)triphenylphosphonium chloride
(0.0062 mol) in THF (20 ml). The mixture was stirred for 15 min. A solution of
intermediate 11 (prepared according Al.k) (0.00514 mol) in THF (20 ml) was added
dropwise. The reaction mixture was stirred for 16 hours at 50 °C. Water was added and
this mixture was extracted with Et2O. The separated organic layer was dried (MgSCU),
filtered and the solvent evaporated in vacuo. The residue was purified by short open
column chromatography over silica gel (eluent: CHkCh/CHsOH 97/3), then by HPLC
(eluent: CH2Cl2/(CH3OH/NH3) 99/1) to separate the (E)/(Z) isomers. Two product
fraction groups were collected and their solvent was evaporated. Yield: 0.651 g of
compound 24 (26%, (E)).
ExampleB 13
Preparation of compound 25
Resin f (0.0016 mol; 1.5 mmole/g) was suspended in THF. 1.6 M BuLi
(0.0015 mol) was added and the mixture was stirred for 15 min. The mixture was
filtered and the filter residue (resin) was washed with anhydrous THF (3 x). The resin
was suspended in THF (5 ml). Intermediate 19 (prepared according to Al.s) (0.0004
mol) was added and the reaction mixture was stirred overnight at 100 °C. The mixture
was cooled, filtered and the filtrate was evaporated in vacuo. The residue was purified
by preparatory HPLC (eluent: CH2Cl2/(CH3OH/NH3) 97/3). The product fractions
were collected and the solvent was evaporated. Yield: 0.168 g of compound 25 .
Example B14
Preparation of compound 26
A mixture of intermediate 16 (prepared according to Al.p) (0.0025 mol), benzenamine
(0.0028 mol) and NaBH, (0.0028 mol) in HOAc (50 ml) was stirred for 2 hours at
room temperature. An aqueous NHiOH solution was added. This mixture was extracted
with CH2Ch. The separated organic layer was dried (MgSO4), filtered and the solvent
evaporated in vacuo. The residue was purified by short open column chromatography
over silica gel (eluent: CH2Cl2/(CH3OH/NH3) 99/1), then by flash column
chromatography over silica gel (eluent: CH2Cl2/(CH3OH/NH3) 98/2). The product
fractions were collected and the solvent was evaporated. Yield: 0.181 g of compound
26 (15%).
Example B15
Preparation of compound 27
(0.0018 mol) was added dropwise to a solution of intermediate
17 (prepared according to Al.q) (0.0012 mol), 3-fluorophenol (0.0018 mol) and PPh3,
pol. (0.0024 mol) in THF, dry (10 ml), under N2 atmosphere. The reaction mixture was
stirred overnight at room temperature. The mixture was filtered, washed with CH^Ch
and CH3OH, and the solvent was evaporated. The residue was purified by short open
column chromatography over silica gel (eluent: CH2C12/EtOAc 2/1 and
96/4), then by flash column chromatography over silica gel (eluent:
97/3). The product fractions were collected and the solvent was evaporated. Yield:
0.29 g of compound 27 (50%).
Example B16
Preparation of compound 28
NaClO (4%) (0.005 mol) was added to a solution of intermediate 35 (prepared
according A4.d) (0.002 mol) in CH2C]2 (10 ml). The reaction mixture was stirred for 4
hours at room temperature. Et3N (0.004 mol) was added and the reaction mixture was
stirred for 24 hours at room temperature. The organic layer was separated, washed with
Na2SCO3 (10%), dried (Na2SCU), filtered and the solvent was evaporated. The residue
was purified by short open column chromatography over silica gel (eluent: EtOAc and
CH2Cl2/CH3OH 95/5). The desired fractions were collected and the solvent was
evaporated. The residue was taken up into diethyl ether, then filtered through dicalite
and the filtrate was evaporated. The residue was dissolved in diethyl ether and
converted into the hydrochloric acid salt (1:2). The precipitate was filtered off, washed
with 2-propanone and diethyl ether, and dried. Yield: 0.15 g of compound 28 (15%).
ExampleB 17
Preparation of compound 29
A mixture of intermediate 37 (prepared according to A5.g) (0.006 mol) and 2-
(bromomethyl)naphthalene (0.003 mol) in dioxane (40 ml) was stirred at 100 °C for 6
hours, then overnight at room temperature. The reaction mixture was treated with a
10% aqueous K2CO3 solution, then extracted with CHjC^. The separated organic layer
was evaporated. The residue was purified by short open column chromatography over
silica gel (eluent: CH2CI2/CH3OH 95/5 and 90/10 and CH2Cl2/(CH3OH/NH3) 95/5).
The product fractions were collected and the solvent was evaporated. The residue (1.49
g) was treated with diethyl ether, then dried. Yield: 0.8 g of compound 29 (53%).
In the following tables (Tables 1-5) a number of compounds are given which have
been prepared according to any one of the Examples above. All compounds have also
been tested for their pharmacological activity.
(Table Removed)
C. Pharmacological examples
Example Cl : Binding experiment for qadrenergic receptor subtypes and for
5-HT transporter
General
The interaction of the compounds of Formula (I) with ha2-receptors and H5-HTtransporters
was assessed in in vitro radioligand binding experiments. In general, a low
concentration of a radioligand with a high binding affinity for a particular receptor or
transporter is incubated with a sample of a tissue preparation enriched in a particular
receptor or transporter or with a preparation of cells expressing cloned human receptors
in a buffered medium. During the incubation, the radioligand binds to the receptor or
transporter. When equilibrium of binding is reached, the receptor bound radioactivity
is separated from the non-bound radioactivity, and the receptor- or transporter-bound
activity is counted. The interaction of the test compounds with the receptor is assessed
in competition binding experiments. Various concentrations of the test compound are
added to the incubation mixture containing the receptor- or transporter preparation and
the radioligand. The test compound in proportion to its binding affinity and its
concentration inhibits binding of the radioligand. The radioligand used for hcx2A>
hcc2B and ha-2C receptor binding was [3Hj-raulwolscine and for the h5-HT transporter
was [3H]paroxetine.
Cell culture and membrane preparation.
CHO cells, stabile transfected with human adrenergic-cx2A-, -WIB or OL-IC receptor cDNA,
were cultured in Dulbecco's Modified Eagle's Medium (DMEM)/Nutrient mixture
Ham's F12 (ratio l:l)(Gibco, Gent-Belgium) supplemented with 10 % heat inactivated
fetal calf serum (Life Technologies, Merelbeke-Belgium) and antibiotics (100 lU/ml
penicillin G, 100 /ig/ml streptomycin sulphate, 110 g/ml pyruvic acid and 100 jig/ml
L-glutamine). One day before collection, cells were induced with 5 mM
sodiumbutyrate. Upon 80-90 % of confluence, cells were scraped in phosphate
buffered saline without Ca2"1" and Mg2+ and collected by centrifugation at 1500 x g for
10 min. The cells were homogenised in Tris-HCl 50 mM using an Ultraturrax
homogenizer and centrifuged for 10 min at 23,500 x g. The pellet was washed once by
resuspension and rehomogenization and the final pellet was resuspended in Tris-HCl,
divided in 1 ml aliquots and stored at -70°C.
Binding experiment for cfe-adrenergic receptor subtypes
Membranes were thawed and re-homogenized in incubation buffer (glycylglycine 25
mM, pH 8.0). In a total volume of 500 n\, 2-10 /ig protein was incubated with
[3H]raulwolscine (NET-722) (New England Nuclear, USA) (1 nM final concentration)
with or without competitor for 60 min at 25°C followed by rapid filtration over GF/B
filter using a Filtermatel96 harvester (Packard, Meriden, CT). Filters were rinsed
extensively with ice-cold rinsing buffer (Tris-HCl 50 mM pH 7.4). Filter-bound
radioactivity was determined by scintillation counting in a Topcount (Packard,
Meriden, CT) and results were expressed as counts per minute (cpm). Non-specific
binding was determined in the presence of 1 M oxymetazoline for h(X2A- and
receptors and 1 /iM spiroxatrine for hccac receptors.
Binding experiment for 5-HT transporter
Human platelet membranes (Oceanix Biosciences Corporation, Hanover, MD, USA)
were thawed, diluted in buffer (Tris-HCl 50 mM, 120 mM NaCl and 5 mM KC1) and
quickly (max 3 s) homogenised with an Ultraturrax homogenizer. In a total volume of
250 uL, 50-100 ug protein was incubated with [3H]paroxetine (NET-869) (New
England Nuclear, USA) (0.5 nM final concentration) with or without competitor for 60
min at 25 °C . Incubation was stopped by rapid filtration of the incubation mixture over
GF/B filters, pre-wetted with 0.1 % polyethyleneamine, using a Filtermatel96 harvester
(Packard, Meriden, CT). Filters were rinsed extensively with ice-cold buffer and
radioactivity on the filters was counted in a Topcount liquid scintillation counter
(Packard, Meriden, CT). Data were expressed as cpm. Imipramine (at 1 p.M final
concentration) was used to determine the non-specific binding.
Data analysis and results
Data from assays in the presence of compound were calculated as a percentage of total
binding measured in the absence of test compound. Inhibition curves, plotting percent
of total binding versus the log value of the concentration of the test compound, were
automatically generated, and sigmoidal inhibition curves were fitted using non-linear
regression. The pICso values of test compounds were derived from individual curves.
All compounds according to formula (I) produced an inhibition at least at the htt2A site
(but often also at the htt2B and ha2C sites) and simultaneously at the 5-HT transporter
site of more than 50 % (pICso) at a test concentration ranging between
1(H-6 M and 10~9 M in a concentration-dependent manner. For a selected number of
compounds, covering most of the various embodiments of Formula (I), the results of
the in vitro studies are given in Table 6.
Table 6 : Some results of the in vitro experiments (pICso-values). n.d.: not determined.
Comp
(Table Removed)
Example C2 : In vivo experiment for O-adrenoceptor antagonism and for serotonine
(5-HT) reuptake inhibition activity.
A. Medetomidine-test
The onset and end of medetomidine (0.10 mg/kg, i.v.)-induced loss of righting was
recorded in overnight-starved Wiga male rats (200-250 g). The intensity of the loss of
righting was scored: 0 = normal behaviour, 1 = slight ataxia, 2 = pronounced ataxia,
3 = loss of righting for a period 5 min.
Under standard conditions, test compound or solvent was administered (s.c. or p.o.) 1 h
before medetomidine. Criterion for drug-induced antagonism: (1) antagonism of loss
of righting: duration = 0 min (1.4% false positive controls; n = 74) (2) reversal of
ataxia: score righting reflex over a period longer than 120 min (0% false positives). Centrally acting
tti-adrenoceptor antagonists or behavioural stimulants antagonise the loss of righting ;
sedative compounds may result in prolongation.
The following observations were made : onset of loss of righting (min), end of loss of
righting (min) and intensity of loss of righting (score 0-4). The observations were
performed at 1 h following s.c. (solutions) or p.o. (suspensions) administration,
respectively. Starting dose was 10 mg/kg (References : Berger, U.V., Grzanna, R.,
Molliver, M.E., Exp. Neurol. 103, 111-115 (1989), Fuller, R.W., Perry, K.W., Molloy,
B.B.. Eur. J. Pharmacol. 33, 119-124 (1975) and Lassen, B.J., Psvchopharmacol. 57,
151-153(1978)).
B. pCA-test
Male Wiga rats were used (body weight: 200 ± 20 g). One hour after administration of
test compound or solvent, a solution of pCA was injected subcutaneously (5 mg/kg ; 10
ml/kg). Forty-five minutes after the pCA injection, head-twitches (HTW) are counted
and the excitation (EXC) were scored over three successive 5 min intervals (starting 45,
50 and 55 minutes after pCA-administration. The scores were given by a trained
observator according to the intensity scale: 0 = absent or doubtful, 1 = present,
2 = pronounced, 3 = maximal. For statistical analysis, the head-twitches counted during
the 15-min observation time were cumulated. For the other phenomena, the median
value of the three 5-min-intervals was used.
Standard observations were performed at 1 h following s.c.or p.o. administration. The
starting dose was in general 10 mg/kg. Doses were initially given to 2 animals. When
both animals show activity for at least one of the observations, the compound was
considered active and testing was repeated at a 4 times lower dose. When activity was
found in only one out of the two animals, an additional animal was tested. When
activity was found in this additional animal, the compound was also considered to be
active and testing was repeated at a 4 times lower dose. In all other cases the
compound was considered inactive at the particular time-route-dose (Reference :
Janssen, P.A.J., Niemegeers, C.J.E., Awouters, F., Schellekens, K.H.L., Megens,
A.A.H.P., Meert, T.FJ. Pharmacol. Exp. Thorn. 244. 685-693 (1988)).
Results
A large number of compounds according to the invention showed a central activity
(minimal effective dose) both in the medetomidine test and in the pCA-test of less than
or equal to 10 mg/kg.
Example C3 : r3SS1GTPyS binding assay
Membranes of the hccaA adrenoceptor expressing CHO cell line were thawed and rehomogenised
in 20 mM Hepes buffer. The incubation medium consisted of: 20 mM
Hepes buffer, pH 7.5,1 jiM GDP, 3 mM MgCl2, 100 mM NaCl, 0.25 nM [35S]GTPyS
and 10 u.g protein per well of a 96-well plate. Antagonists and the reference agonist
noradrenaline (3 pM) were added 20 min before the[35S]GTPyS. The incubation (20
min, 37°C) was ended by rapid filtration through GF/B filters and binding quantified by
liquid scintillation counting.
Results
All compounds according to the invention evaluated in the GTPyS binding assay did
not show significant increases of [35S]GTPyS binding to the ha2A receptor up to 10
uM. All compounds evaluated in the assay were able to inhibit noradrenaline-induced
increases of [35S]GTPyS binding, thereby showing their antagonistic nature at this
receptor.

We claim
1. Isoxazoline compounds of general Formula (I)
(Formula Removed)

the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the N-oxide form thereof, wherein :
X is CH2, N-R7, S or O;
R7 is selected from the group of hydrogen, alkyl, phenyl,
phenylalkyl, alkylcarbonyl, alkyloxycarbonyl and mono- and dialkylaminocarbonyl, the phenyl and alkyl groups being optionally substituted with one or more halo atoms;
R1 and R2 are each, independently from each other, selected from the
group of hydrogen, hydroxy, cyano, halo, OSO2H, OSO2CH3,
phenyl, phenylalkyl, alkyloxy, alkyloxyalkyloxy,
alkyloxyalkyloxyalkyloxy, tetrahydrofuranyloxy, alkylcarbonyl-oxy, alkylthio, alkyloxyalkylcarbonyloxy, pyridinylcarbonyloxy, alkylcarbonyloxyalkyloxy, alkyloxycarbonyloxy, alkenyloxy, alkenylcarbonyloxy and mono-and dialkylaminoalkyloxy, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; or

R1 and R2 may be taken together to form a bivalent radical -R1-R2- selected from the group of -CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-O-, -CH2-O-CH2- and -O-CH2-CH2-O- ;
a and b are asymmetric centers ;
(CH2)m is a straight hydrocarbon chain of m carbon atoms, m being
an integer ranging from 1 to 4 ;
Pir is a radical according to any one of Formula (IIa), (IIB or (IIc)
(Formula Removed)

wherein :
each R8 is independently from each other, selected from
the group of hydrogen, hydroxy, amino, nitro,
cyano, halo and alkyl ;
n is an integer ranging from 1 to 5 ;
R9 is selected from the group of hydrogen, alkyl and
formyl;
R3 is a radical according to any one of Formula (III)
(Formula Removed)

wherein :
d is a single bond while Z is a bivalent radical selected from the group of -CH2-, -C(=O)-, -CH(OH)-, -C(=N-OH)-, -CH(alkyl)-, -O-, -S-, -S(=O), -NH- and -SH-; or d is a double bond while Z is a trivalent radical of formula =CH- or =C(alkyl);
A is a 5- or 6-membered aromatic homocyclic or heterocyclic ring, selected from the group of phenyl, pyranyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl, isothiazolyl, pyrrolyl, imidazolyl, pyrazolyl, furanyl, oxadiazolyl and isoxazolyl;
p is an integer ranging from 0 to 4 ;
q is an integer ranging from 0 to 7 ;
R4 is selected from the group of hydrogen, alkyl, phenyl, biphenyl, naphthyl, halo and cyano, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ;
R5 is equal to R4 ; or
R4 and R5 may be taken together to form a bivalent radical -R4- R5- selected from the group of -CH2-, =CH,-CH2-CH2,-CH=CH-,-O-,-NH-,=N-,-S-,-CH2N (-alkyl, -CH=N -CH2O- and -OCH2- ;
each R6 is independently from each other, selected from the group of hydrogen, hydroxy, amino, nitro, cyano, halo, carboxyl, alkyl, phenyl, alkyloxy,

phenyloxy, alkylcarbonyloxy, alkyloxycarbonyl,
alkylthio, mono- and dialkylamino,
alkylcarbonylamino, mono- and
dialkylaminocarbonyl, mono- and dialkyl-aminocarbonyloxy, mono- and dialkyl-aminoalkyloxy, the alkyl and aryl radicals being optionally substituted with one or more hydroxy or halo atoms or amino groups ; or
two vicinal radicals R6 may be taken together to form a
bivalent radical -R6-R6- selected from the group
of -CH2-CH2-O-, -O-CH2-CH2-, -O-CH2-C(=O),-O-
CH2-O-, -CH2 -O- CH2-,-O-CH2 -CH2
-0-, -CH=CH-CH=CH-,-CH=CH-CH=N-, -CH=CH-
N=CH-,-CH= N-CH=CH-, -N=CH-CH=
CH-, -CH2-CH2-CH2-, -CH2-CH2-C(=O)-
and -CH2-CH2-CH2-CH2- ; and
R10 is selected from the group of hydrogen, alkyl,
phenylalkyl and phenyl.
2. A compound as claimed in claim 1, wherein X=O or NH ; R1 and R2 are both alkyloxy ; m = 1 ; Pir is a radical according to Formula (IIa) wherein R8 is hydrogen and n = 4 ; R3 is a radical according to Formula (IIIb) wherein Z is =CH-, d is a double bond, A is a phenyl ring, R4 is an alkyl and R10 is hydrogen .
3. A compound as claimed in any one of claims 1-2 for use as a medicine.

4. A prodrug of a compound of Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the iV-oxide form thereof, which is degraded in vivo to yield a compound according to Formula (I), the pharmaceutically acceptable acid or base addition salts thereof, the stereochemically isomeric forms thereof and the iV-oxide form thereof.
5. A process for preparing a compound as claimed in any one of claims 1-2, wherein a compound according to Formula (IV) is reacted with a compound according to Formula (V) or Formula (VII).
6. A compound of general Formula (VIII)
(Formula Removed)

wherein R1, R2, X, m, R8 and n are defined as in Formula (I).

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Patent Number

234574

Indian Patent Application Number

00968/DELNP/2003

PG Journal Number

31/2009

Publication Date

31-Jul-2009

Grant Date

09-Jun-2009

Date of Filing

24-Jun-2003

Name of Patentee

JANSSEN PHARMACEUTICA N.V,

Applicant Address

TURNHOUTSEWEG 30,2340 BEERSE,BELGIUM,

Inventors:

#	Inventor's Name	Inventor's Address
1	JOSE MARIA CID-NUNEZ	C/O JANSSEN-CILAG,S.A. EDIFICIO JOHNSON & JOHNSON,PASEO DE LAS DOCE ESTRELLAS,5-7,CAMPO DE LAS NACIONES,28042 MADRID,SPAIN,
2	JOAQUIN PASTOR-FERNANDEZ	C/O JANSSEN-CILAG,S.A. EDIFICIO JOHNSON & JOHNSON,PASEO DE LAS DOCE ESTRELLAS,5-7,CAMPO DE LAS NACIONES,28042 MADRID,SPAIN,
3	JOSE IGNACIO ANDRES-GIL	C/O JANSSEN-CILAG,S.A. EDIFICIO JOHNSON & JOHNSON,PASEO DE LAS DOCE ESTRELLAS,5-7,CAMPO DE LAS NACIONES,28042 MADRID,SPAIN,
4	FRANCISCO JAVIER FERNANDEZ-GADEA	C/O JANSSEN-CILAG,S.A. EDIFICIO JOHNSON & JOHNSON,PASEO DE LAS DOCE ESTRELLAS,5-7,CAMPO DE LAS NACIONES,28042 MADRID,SPAIN,
5	MANUEL JESUS ALCAZAR-VACA	C/O JANSSEN-CILAG,S.A. EDIFICIO JOHNSON & JOHNSON,PASEO DE LAS DOCE ESTRELLAS,5-7,CAMPO DE LAS NACIONES,28042 MADRID,SPAIN,
6	ANTONIUS ADRIANUS HENDRIKUS PETRUS MEGENS	C/O JANSSEN PHARMACEUTICA N.V.TURNHOUTSEWEG 30,2340 BEERSE,BELGIUM,
7	GODELIEVE IRMA CHRISTINE MARIA HEYLEN	C/O JANSSEN PHARMACEUTICA N.V.TURNHOUTSEWEG 30,2340 BEERSE,BELGIUM,
8	XAVIER JEAN MICHEL LANGLOIS	C/O JANSSEN PHARMACEUTICA N.V.TURNHOUTSEWEG 30,2340 BEERSE,BELGIUM,
9	MARGARETHA HENRICA MARIA BAKKER	C/O JANSSEN PHARMACEUTICA N.V.TURNHOUTSEWEG 30,2340 BEERSE,BELGIUM,
10	THOMAS HORST WOLFGANG STECKLER	C/O JANSSEN PHARMACEUTICAL N.V.TURNHOUTSEWEG 30,2340 BEERSE,BELGIUM,

PCT International Classification Number

C07D 498/04

PCT International Application Number

PCT/EP02/01567

PCT International Filing date

2002-02-13

PCT Conventions:

#	PCT Application Number	Date of Convention	Priority Country
1	01200611.0	2001-02-21	EUROPEAN UNION