ISOXAZOLES WITH NOOTROPIC ACTIVITY

Abstract: Compounds with nootropic activity of formula (I) (wherein R, R1 and n have the meanings reported in the specification), pharmaceutically acceptable salts thereof, processes for their preparation and their therapeutic use are described.

Description: Isoxazoles with nootropic activity. The present invention relates to compounds with nootropic activity and, more particularly, it relates to derivatives of N-(5-isox- azolecarbonyl)-4-aπ_inobutyric acid, the processes for their prep¬ aration, their therapeutic use and the pharmaceutical compositions containing them as active ingredient. Piracetam, 2-pyrrolidinoneacetamide (Merck Index, X edition, page lθ8θ, no. 7363) is a compound described in Belgian patent no. 667906 (Union Chimique Beige) as central stimulant. For its action at cerebral level on glucose metabolism as well as, especially, in increasing acεtylcholine release, Piracetam is considered the parent compound of nootropic drugs and it is used in therapy in the treatment of cerebral efficiency disorders, particularly in elderly patients. Piracetam, even if it is used in therapy, shows a relative eff¬ ectiveness. In order to improve the therapeutic activity of the parent com¬ pound several different compounds have been prepared. Among these we can mention, for example, the derivatives of 1- benzoyl-2-pyrrolidinone described in European patent no. 5143 (Hoffmann-La Roche) and, particularly, the derivative known as Aniracetam, l-(4-nιethoxy-benzoyl.-2-pyrrolidinone (USAN and the USP Dictionary of Drug Names 1987, page 30). It is worth noting, however, that, as far as we know, the com¬ pounds with nootropic activity developed up to now do not contain an isoxazole ring and furthermore most of them have a remarkable structural analogy with the parent compound Piracetam. Besides, as can be seen also in Aniracetam, the pyrrolidinone ring remains present, as peculiar characteristic. We have surprisingly found and they are the object of the present invention, compounds of formula wherein n represents a number selected from 1 and 2j R represents a hydrogen atom, a halogen atom, hydroxy, a C -C, alkyl or alkoxy; R rep alkoxy or 1 an R en atom, a linear or branched C -C, alkyl, a group of formula (CH ) R_ l o 2 m 4 wherein m represents an integer selected from 1, 2 and 3 . represents a carboxy group, an alkoxycarbonyl group having from 1 to 6 carbon atoms in the alkoxy moiety or a group of I formula R ^ R , R_ and R,, t e same or different, represent a hydrogen atom or 5 o a linear or branched C -C, alkyl ; or one of R and R represents a group of formula R -CH-R wherein R has the above reported meanings and R represents a linear or branched C -C alkyl optionally substituted by phenyl, hydroxy or by a mercapto group; and the other repre¬ sents a hydrogen atomj or, in addition, R and R - together with the nitrogen atom to which they are bonded, form a 5, 6 or 7 membered heterocycle which may further contain 1 or 2 heteroatoms selected among nitrogen, oxygen and sulphur. Specific examples are 1-pyrrolidinyl, 1-piperazinyl, 4-πιethyl- 1-piperazinyl, 1-piperidyl and l-pyrrolidinyl-2-one. A further object of the present invention are the salts of the compounds of formula I, which have an acidic function, with pharmaceutically acceptable organic or inorganic bases and the salts of the compounds of formula I, wherein R contains a basic function, with organic or inorganic acids suitable for pharmaceu¬ tical use. Examples of organic or inorganic bases useful as salifying agents are sodium or potassium hydroxides, ammonium hydroxide, lysine, arginine, cysteine and 2-amino-2-hydroxymethyl-l,3-propanediol. Examples of organic or inorganic acids useful as salifying agents are hydrochloric acid, hydrobromic acid, sulfuric acid, tartaric acid, citric acid and glutamic acid. The compounds of formula I, object of the present invention, have nootropic activity. Examples of compounds comprised in formula I are the following: X-C-NH-CH -CH -CH -C00H i l 2 2 2 X-C-NH-CH -CH -CH -C00CH ,, i| 2 2 2 3 X-C-NH-CH -CH -CH -COOC H „ II 2 2 2 2 5 0 - 4 - X-C-NH-CH -CH -CH -C-N N-CH, II 2 2 2 I. \ / X-C-NH-CH -CH -CH -COOC H I! 2 2 2 49 X-C-NH-CH -CH -CH -C-NH-CH -CH -CH -C-MT |j 2 2 2 |j 2 2 2 | | 2 0 0 0 X-C-NH-CH -CH -CH -C-NH II 2 2 2 j| 2 0 0 X-C-NH-CH -CH -CH -C-NH-CH -COOC H _ If 2 2 2 ,| 2 2 5 X-C-_ffi-CH -CH -CH -C-NH-CH -CH -CH -COOC H || 2 2 2 || 2 2 2 2 5 X-C-NH-CH -CH -CH -C-NH-CH -COOC,H II 2 2 2 it 2 6 13 / 49 X-C-_«-CH -CH -CH -C-rø-CH -CH -CH -C-N ii 2 2 2 π 2 2 2 II \ 0 C H 49 X-C-NH-CH -CH -CH -C-NH -CH -CH -CH -C-N NH-HC1 li 2 2 2 II 2 2 2 || \ / X-C-NH-CH_-CH_-CH_-C-NH-CH„-CH o -CH o -C00-CH o -(CH„) -(CH=CH-CH_) -CH || 2 2 2 || 2 2 2 2 2 7 2 3 3 0 0 X-C-rø-CH -CH -CH -C00-CH -(CH ) . , -CH. || 2 2 2 2 2 14 3 o X-C-NH-CH -CH -CH -C-_ffl-CH -CH -CH -COOH II 2 2 2 ii 2 2 2 X-C-NH-CH -CH -CH -C-NH-CH -COOH li 2 2 2 II 2 wherein X represents the group of formula R N wherein R has the above reported meanings but preferably it represents a chlorine or bromine atom, hydroxy, methoxy or ethoxy, methyl or ethyl. The preparation of the compounds of formula I is another object of the present invention and comprises a condensation reaction between a chloride of 5-isoxazolecarboxylic acid of formula wherein R has the above reported meanings, and 4-aminobutyric acid or a derivative thereof of formula H-(NH-CH -CH -CH -CO) -R, (II-A) 2 2 2 n 1 wherein n and R have the above reported meanings, in an inert solvent in the presence of an acceptor of halogenhy- dric acids. Examples of suitable acceptors of halogenhydric acids are inorgan- ic or organic bases such as sodium hydroxide or carbonate, trieth- ylamine, N-methyl-morpholine and pyridine. The reaction of compounds II with 4-aminobutyric acid (II-A n=l R -sOH) gives the compounds of formula I wherein n=l and R =0H. Such compounds are transformed into their reactive derivatives - such as mixed anhydrides and, without any isolation, they are condensed with 4-aminobutyric acid giving the compounds of formula I wherein n=2 and R =0H. For greater simplicity the compounds of formula I wherein R =QH will be indicated hereinafter as compound I-a) The compounds of formula I wherein R is alkoxr are prepared from compounds la by esterification. The esterification reaction is carried out with saturated or unsaturated aliphatic alcohols such as me hanol, ethanol, propa- nol, butanol, cetylic alcohol, linolenic alcohol or stearic alcohol, in the presence of catalytic amounts of an inorganic by reacting suitable reactive derivatives of compounds la with a suitable amino derivative according to the following reaction wherein , R, R , R and R have the above reported meanings and Y repre- 1 — o sents a leaving group. Examples of reactive derivatives (III) of the compounds of formula la are mixed anhydrides obtainable by reaction with alkylchloro- formates such as isobutylchloroforraate. Compounds III are, then, reacted, without any isolation, with an amino derivative of formula IV in a suitable solvent, which may be compound IV itself. Examples of suitable amino derivatives of formula IV are ammonia, mono- or di-substituted amines such as methylamine, dimethylamine, diethylamine and di-n.propylamine; alkylendiamines such as N,N- diethyl-ethylendiamine and N,N-diisopropyl-ethylendiamine; cyclic derivatives such as pyrrolidine, 2-pyrrolidinone, piperazine and N-methyl-piperazine or natural aminoacids such as glycine, ala- nine, valine, serine, cysteine, phenylalanine and derivatives thereof. The comnounds of formula I wherein n=l and the ^N group is a 2 3 l-pyrrolidinyl-2-one may also be prepared from compounds la (n=2) by a cyclization reaction. The cyclization reaction is carried out in one step through the formation of the corresponding acyl halide as intermediate. This intermediate, without any isolation, cyclizes in the same I reaction environment giving the compound I wherein n=l and ^ 2 3 is a l-pyrrolidinyl-2-one. The intermediate acyl halides are prepared by reaction with a suitable halogenating agents such as thionyl chloride, phosphorus trichloride, phosphorus pentachloride, phosphorus tribromide, phosphorus pentachloride, oxalyl bromide and oxalyl chloride. For economical and practical reasons only, thionyl chloride is preferably used. The cyclization reaction is carried out optionally in the presence of inert organic solvents such as toluene and benzene. It is clear to the man skilled in the art that the compounds of formula I, object of the present invention, can be prepared following alternative routes too. For example the esterification of the carboxyl group can be carried out directly on 4-aminobutyric acid or a derivative thereof, that is before condensation with compound II. In a similar way the compounds of formula I wherein R = n^ _- __ can 1 °_ ^ R. be prepared also according to the following- reaction 2 3 4- H-(NH-CH (ii) wherein n, R, R , R and R have the above reoorted meanings, ~ 1 2 Λ According to this method, at first, the functionalization of the carboxyl group of 4-aminobutyric acid or of 4-(4-a-πιinobutyroyl)- aminobutyric acid is carried out in order to obtain the amides of 5 formula V. These amides are, then, condensed with the acyl chloride of 5- isoxazolecarboxylic acid (II), in an inert organic solvent and in , to give the 0 The compounds of formula I are active on central nervous system and they can be used in pharmaceutical field as nootropics. By the electroshock (ECS) amnesia test, described in example 8, in rat the pharmacological activity of the compounds object of the present invention was evaluated in comparison with the activity of 5 Piracetam and Aniracetam. The test results showed that the ED values (umol/kg) of the compounds of formula I are generally remarkably lower than that of Piracetam and Aniracetam. Particularly, the ED values are up to 20 times lower in case of 0 "per os" administration and up to 10 times lower by perfusion than that of Piracetam. The compounds of formula I have no toxicological problem. They exhibit a complete tolerability even when they are adminis¬ tered by perfusion at dose of 2 g/kg. EC The therapeutic uses of the compounds object of the present invention are in the treatment of impairments of cerebral func¬ tionality due to ageing as well as to pathologic or traumatic reasons such as for example amnesia and decrease in cognitive capacity. The therapeutic dose cf the compounds cf formula I and of the salts thereof depends on several factors such as the way of administration, the specific pharmaceutical composition, the treatment needed and the individual response to the therapy. In general it will be comprised between 5 and 2000 mg/day in one or more administrations. A further object of the present invention are the pharmaceutical compositions containing the compounds of formula I or pharmaceuti- cally acceptable salts thereof as. -active ingredient optionally together with one or more, solid or liquid, organic or inorganic pharmaceutical excipients such as diluents, preserving agents, moistening agents, colouring agents, flavouring agents and so on. The pharmaceutical compositions object of the present invention can be administered in solid pharmaceutical preparations, such as tablets, coated tablets, capsules, granulates and suppositories or in liquid pharmaceutical preparations such as syrups, suspensions, emulsions and solutions suitable for oral or parenteral adminis¬ tration. The compounds object of the present invention can be prepared in slow and protracted release pharmaceutical formulations too. The pharmaceutical compositions object of the present invention may also contain the compounds of formula I or pharmaceutically acceptable salts thereof in association with other active ingredi¬ ents selected, for example, among aminoacids, N-acetyl-cysteine, co-enzymes, mineral salts and vitamins. The preparation of the pharmaceutical compositions object of the present invention is carried out by usual techniques. In order to better illustrate the present invention, the following examples are now given. Example 1 Preparation of N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyric acid (Compound no. 1) To a solution of 4-aminobutyric acid (11.34 g . 0.11 mol) and sodium hydroxide (4-4 g > 0.11 mol) in water (150 ml), cooled at 10°C, a solution of 3-b ~ omo-5-isoxazolecarbonylchloride (21 g; 0.10 mol) in toluene (150 ml) and a solution of sodium hydroxide (4«0 ; 0.10 mol) in water (150 ml) were added simultaneously. The reaction mixture was kept under stirring at room temperature for 2 hours. The aqueous layer was separated, washed with toluene (150 ml) and acidified with concentrated hydrochloric acid up to pH 2. The precipitate was extracted three times with ethyl acetate (200 ml). The organic extracts were collected, washed with water and dried on sodium sulphate. After evaporation of the solvent, the solid crude was cristallized from acetonitrile (150 ml) giving N-(3-bromo-5-isoxazolecarbonyl)- -4-aminobutyric acid (23 g; 83 yield) with m.p. 140-141°C. ^-NMR (DMSO-d : delta (ppm): 1.6-2.1 (2H, m, CH -CH -CH ) 3 2.1- 0 2 — 2 2.5 (2H, m, CH CO); 3.33 (2H, q, CH N); 7-39 (1H, s, -CH=). In a similar way the following compounds were prepared. N-(3-chloro-5-isoxazolecarbonyl)-4-aminobutyric acid (Compound no. 2) m.p. 140-141-C (acetonitrile); 8l._ yield ^- MR (DMS0-d.): delta (ppm): 1-5-2.1 (2H, m, CH -CH -CH ); 2.1- O 2 — 2 2 2.6 (2H, m, CH CO), 3-36 (2H, q, CH_N); 7-39 (1H, s, -CH--) . N-(3-meτhoxy-5-isoxazolecarbonyl)-_l-aminobuτyric acid (Compound no. 3) m.p. 134-135 0 C (acetonitrile); 787- yield 1 H-NMR (DMS0-d : delta (ppm): 1.5-2.1 (2H, m, CH_-CH_-CH 9 ), ■ 2.1- 2.5 (2H, m, CH_C0) ; 3-29 (2H, q, CH_N) ; 3-97 (3H, s, CH^) ; 6.82 (1H, s, -CH=) . N-( -πιethyl-5— isoxazolecarbonyl)-4-aminobutyric acid (Compound no . 5 *> m.p. 136-137°C (acetonitrile) 75% yield l-NMR (DMSO-d,): delta (ppm): 1.5-2.1 (2H, m, CH -CH -CH ); 2.1- 2.5 (2H, , CH 2 C0); 2.32 (3H, s, CH ); 3-29 (2H, q, O^N); 7-00 (IH, s, -CH=). Q N-(3-n.butyl-5-isoxazolecarbonyl)-4-aminobutyric acid (Compound no. 5) m.p. 130-131-C (acetonitrile); 75 yield 1 H-NMR (DMS0-d 6 ): delta (ppm): 0.90 (3H, t, CH ); 1.32 (2H, , CH -CO); 5 2.26 (2H, dd, CH CO); 2.66 (2H, dd, CH -C=); 3-25 (2H, m, X-CH ); 6.96 (IH, s, -CH=); 8.92 (IH, s, NH); 12.12 (IH, s, OH). N-(5—isoxazolecarbonyl)-4-aminobutyric acid (Compound no. 6) m.p. 149-151°C (acetonitrile); 64?. yield ∑ H-NMR (DMS0-d 6 ): delta (ppm): 1.75 (2H, m, CH 9 -CH -CH ; 2.27 Q (2H, t, CH CO); 3-27 (2H, dt, N-CH ); 7-06 (IH, d, -CH=C); 8.74 (IH, d, -CH=N); 8.99 (IH, s, NH); 12.12 (IH, s, OH). Example 2 Preparation of ethyl N—(3-bromo-5-isoxazolecarbonyl)—4-aminobutyr- ate (Compound no. 7) 5 A suspension of N-(3—bromo—5—is ~ xazαlecarbonyl)-4-aminobutyric acid (10 g; O.O3O mol), prepared according to the method described in example 1, in absolute ethanol (300 ml), containing a catalytic amount of concentrated H SO , was kept under stirring at room 2 4 temperature for 24 hours. 0 The obtained solution was evaporated to dryness and the solid residue was dissolved in water (100 ml) and extracted 3 times with ethyl acetate (100 ml). The organic extracts were collected, washed with water, dried on sodium sulphate and evaporated. The solid residue was crystallized from diisopropyl ether (100 ml) obtaining ethyl N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyrate (9-1 g . 835S yield) with m.p. 72-3°C. ^-NMR (DMSO-d : delta (ppm): 1.18 (3H, t, CH ); 1.5-2.2 (2H, m, CH 2 -CH 2 -CH 2 ); 2.2-2.6 (2H, m, CH_C0); 3-32 (2H, q, CH.N); 4-08 (2H, q, CH 2 -CH ); 7-40 (IH, s, -CH=). Example 3 N-/4-oxo-4-(l-pyrrolidinyl)butyl7-3-bromo-5-isoxazolecarboxamide (Compound no. 8) To a solution of N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyric acid (11.8 g; 0.04 mol), prepared according to the method de¬ scribed in example 1, and N-methyl-morpholine (4-41 nil; 0.04 mol) in anhydrous tetrahydrofuran (200 ml), cooled at -15°C, isobutyl- chloroformate (5.22 ml; 0.04 mol) was slowly added. After standing under stirring at -10°C for 15 minutes, a solution of pyrrolidine (3«30 ml; 0.04 mol) in anhydrous tetrahydrofuran (10 ml) was added dropwise to the reaction mixture kept at the same temperature. At the end of the addition it was kept under stirring for 12 hours at room temperature. After evaporation of the solvent, the residue was dissolved in chloroform (150 ml). The solution was washed with HC1 0.1N ( 50 ml), with water, with an aαueous solution of K C0„ and finally 2 j several times with water up to neutral pH. The organic layer was dried on sodium sulphate and, after evapo- ration of the solvent, the solid residue was crystallized from ethyl acetate (100 ml) obtaining N- 4-oxo-4-(l-pyrrolidinyl)bu~ tyl7-3-bromo-5-isoxazolecarboxamide (10.9 gj 82 yield) with m.p. 120-121-C. l-N R (DMSO-d,): delta (ppm): 1.5-2.1 /6H, m, CH -CH -CH , CH - (CH 2 ) 2 -CH 2 _7; 2.1-2.5 (2H, m, CH^O); 3-1-3-7 (6H, m, C^ , CH 2 -N-CH 2 >; 7-46 (IH, s, -CH=). In a similar way the following compounds were prepared: N-/4-(4-methvl-l-oi D erazinvl)-4-oxobutvl7-3-bromo-5-isoxazole- carboxamide (Compound no. 9) m.p. 124-125°C (ethyl acetate); 85 yield ^-NMR (DMSO-d,): delta (ppm): 1.5-2.0 (2H, m, CH -CH -CH ); 2.0- CH„ CO I 3 I 2.5 (9H, , CH 2 C0, CH 2 - -CH 2 ); 3-l-3.7(6H, m, C^N, CH^N-CH^ 7.36 (IH, s, -CH=). N-/4-/2-(diethylamino)ethylaminq7-4-oxobutyl7-3-bromo-5-isoxazole- carboxamide (Compound no. 10) m.p. 8θ-82°C (ethyl acetate); 65% yield ^-NMR (DMSO-d,): delta (ppm): 0.93 (6H, t, 2CH ); 1.5-2.8 .0H, m, (CH 2 ) 2 C0, CH 2 - -(CH 2 -CH 3 ) 2 J; 2.9-3-5 (4H, m, 2CH £ N); 7-37 (IH, s, -CH_=). N-/ -/2-(diisopropylamino)ethylaminq7-4-oxobutyl7-3-bromo-5- isoxazolecarboxamide (Compound no. 11) m.p. 8l-83°C (diisopropyl ether - ethyl acetate); 78$ yield H-NMR (DMSO-d,): delta (ppm): O.96 (12H, d, 4CH ; 1.5-3-6 (12H, 0 j m, 5CH , 2CH); 7.41 (IH, s, -CH=) . Ethyl N- N-(3—bromo-5-isoxazolecarbonyl)-tL-aminobutyroyl7£licinate (Compound no. 12) m.p. 72-73°C (diisopropyl ether); 88$ yield 1 H-NMR (DMSO-d,): delta (ppm): 1.19 (3H, t, CH„); 1.5-2.1 (2H, m, o 3 CR 2~- H 2~ H 2 ) ' ' 2 - 1-2*4 (2H > m > CH 2 CO) 5 3-33 < 2H > > CH^CH^N); 3.87 (2H, d, N-CH -CO); 4.I6 (2H, q, CH -CH ) ; 7-46 (IH, s, -CH=) . ~* — -< ό - Ethyl N-/N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyroyl7-4- a inobutyrate (Compound No. 13) m.p. 121-122-C (ethyl acetate); 89$ yield 1 H-NMR (DMSO-d,): delta (ppm): 1.18 (3H, t, CH ); 1.5-2.6 (8H, m, 2CH 2 -CH 2 C0); 2.8-3-6 (4H, m, 2CH N); 4.09 (2H, q, CH^H ) 5 7-39 (IH, s, -CH=). 4- N-(3-bromo-5-isoxazolecarbonyl)-aminq7-butyramide (Compound no. 14) m.p. 178-179-C (ethanol); 6l$ yield H-NMR (DMSO-d,): delta (ppm): 1.72 (2H, m, CH -CH -CH ); 2.09 0 2 — 2 2 (2H, dd, CH CO); 3-24 (2H, m, N-CH ) 5 6.78 (IH, s, H-N-H); 7-29 (IH, s, H-N-H); 7-34 (IH, s, -CH=); 9-06 (IH, t, NH-CH ) . ___ 4-/_ / N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyroyl^7-aminq7- butyramide (Compound no. 15) m.p. 202-204°C (dimeth lformamide); 7 $ yield H-NMR (DMSO-d ): delta (ppm): 1-58-1-73 (4H, m, 2CH -CH -CH ) 5 2.03 (2H, dd, CH CONH ); 2.10 (2H, dd, CH -CONH); 3-01-3-23 (4H, m, 2NHCH ); 6-73 (lH, s, H-N-H); 7-25 (lH, s, H-N-H); 7-34 (lH,s, -CH=); 7-82-9-06 (2H, s, 2C0NH). n.hexyl N-(3-bromo-5-isoxasolecarbonyl)-4-aminobutyrate (Compound no. 16) m.p. 52-54-C (hexane); 44$ yield H-NMR (DMSO-d,): delta (ppm): 0.86 (3H, t, CH ); 1.2-1.3 / " OH, m, (CH 2 ) CH 7; 1.5-1-6 (2H, m, 0CH -CH 2 ) 5 1-77 (2H, , CH 2 -CH 9 -C0); 2.3166 ((22HH,, tt,, C CHH CC00)) ; ; 33--2288 ((22HH,, m, N-CH 2 ) 5 3-99 (2H, t, OCH 2 ) 5 7.35 (1H,S, -CH=), 9.07 (IH, t, NH) 1-/N-/N—(3-bromo—5-isoxazolecarbonyl)-4-aminobutyroyl7-4-aminobu- tyroyl/pyrrolidine (Compound no. 17) m.p. 157-159°C (dimethylformamide); 53% yield " ' ' H-NMR (DMSO-d,); delta (ppm): 1.5-1-9 (8H, m, 4CH ); 2.10 (2H, dd, CH CO); 2.21 (2H, dd, CH CO); 3-04 (2H, m, NHCH ); 3-2-3-4 (6H, m, 3CH N); 7-35 (IH, s, -CH=); 7-83 (IH, t, CONH); 9-06 (lH, t, CONH). Example Preparation of N-^-(di-n.propylamino)-4-oxobutyl7-3-bromo-5- isoxazolecarbσxamide (Compound no. 18) A solution of 3-t>romo-5-isoxazolecarbonyl chloride (2.95 gj 0.014 mol) in methylene chloride (5 ml) was added dropwise to a solu¬ tion, cooled at 5°C, of 4-aminobutyric acid di-n.propylamide (2.60 g; 0.014 mol) and triethylamine (1.95 ml; 0.014 mol) in methylene chloride (52 ml). At the end of the addition the reaction mixture was kept under stirring for 6 hours at room temperature and then it was extracted with HC1 0.1N (20 ml), with water, with a saturated aqueous solution of NaHCO (20 ml) and finally with water up to neutral pH. The organic layer was dried on sodium sulphate and, after evapo¬ ration of the solvent, the solid residue was crystallized from diisopropylether . N- 4-(di-n_propylamino )-4-oxobutyl7-3-bromo-5-isoxazolecarboxamide ( 3- 6 g 3 71$ yield) was obtained with m.p. 78-79°C. X H-NMR (DMS0-d, ) : delta (ppm) : 0.82 ( 6H, t, 2CH„) . 1.1-2.1 ( OH, m, 2CH CH„, CH -CH -CH ) ; 2.1-2.5 (2H, m, CH C0) ; 3 - 0-3 - 5 ( 6H, , — j — 2 2 2 CH 2 NH , 2CH 2 N) ; 7- 33 ( IH, s , -CH=) . Example 5 Preparation of N-/N-(3-bromo-5-isoxazolecarbonyl)-4-aminobu- tyroyl7-4-aminobutyric acid (Compound no. 19) Method A To a solution of ethyl N- N-(3-bromo-5-isoxazolecarbonyl)-4- aminobutyroyl7-4-aminobutyrate (23-6 g; O.Oόl mol), prepared according to the method described in example 3 . in methanol (236 ml), cooled at 10°C, a solution of sodium hydroxide (2.64 g; O.O65 mol) in water (6θ ml) was added dropwise. At the end of the addition, it was kept under stirring at room temperature for 18 hours. After evaporation of most solvent, the solution was diluted with water (200 ml) and extracted twice with chloroform 50 ml). The aqueous layer was acidified to pH 2 with HC1 at 10$. The precipi- tate was filtered, washed with water up to neutral pH and it was crystallized from isopropanol (200 ml). N-^-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyroyl7-4-a_ι_inobutyric acid was obtained (20.8 5 94$ yield) with m.p. 157-158°C. 1 H-NMR (DMSO-d ): delta (ppm): 1.4-2-5 (8H, m, 2CH -CH -C0) } 2.8- 3-6 (4H, m, 2CH N); 7-37 (IH, s, -CH=) . In a similar way the following compound was prepared N-/N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyroyl7glycine (Com¬ pound no. 20) m.p. I87-I89-C (isopropanol); 91$ yield 1 H-NMR (DMSO-d ): delta (ppm): 1.5-2.0 (2H, m, CH -CH -CH ) , 2.0- O __. -_. _ 2.5 (2H, m, CH -CH -CO); 3-0-3-6 (2H, m, CH -CH N) } 3-80 (2H, d, CH C00), 7-43 (IH, s, -CH--). Method B To a solution of N-(3-bromo-5-isoxazolecarbonvl)-4-aminobutyric acid (69.27 g; 0.25 mol), prepared according to the method de- scribed in example 1, and triethylamine (34.80 ml; 0.25 mol) in anhydrous tetrahydrofuran (700 ml), cooled at -15°C, isobutylchlo- roformate (32.67 ml, 0.25 mol) was added slowly. After stirring at -10°C for 15 minutes, a solution of 4-aminobu¬ tyric acid (30.94 g; 0.30 mol) and triethylamine (41-76 ml; 0.30 mol) in water (190 ml) and dimethylformamide (60 ml) was added dropwise keeping the reaction mixture at the same temperature. After stirring at room temperature for 3 hours, the solvent was evaporated and the residue was dissolved in water (350 ml). The obtained aqueous solution was acidified with HC1 at 10$ and the precipitate was filtered, washed with water up to neutral pH and it was crystallized from isopropanol (600 ml). N- N-(3-bromo-5-isoxazolecarbonyl)-4-aminobut ~~ roy^-4-aminobutyric acid was obtained (55- 5 g; 6l$ yield) with m.p . 157-158°C . Example 6 Preparation of l-/N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyr- oyl7-2-pyrrolidinone (Compound no. 21) A solution of l- N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyr- oy^-4-aminobutyric acid (9-0 g; 0.025 mol), prepared according to the method described in example 5 . and thionyl chloride (2.72 ml; 0.0375 mol) in toluene (250 ml) was heated under reflux for 15 minutes. After cooling and evaporation of the solvent, the solid residue was cristallized from absolute ethanol (60 ml) giving l-/N-(3- bromo-5-isoxazolecarbonyl)-4-aminobutyrαrl7-2-pyrrolidinone (7-6 g; 88$ yield) with m.p. 141-142°C. L H-NMR ( DMS0-d, ) : delta (ppm) : 1.6-2.3 (4H, m, 2CH -CH -CH ) , O 2 — 2 2 - 3-4-0 (8H, m, 2CH N, 2CH 9 C0) ; 7-41 ( IH, s , -CH=) . Example 7 N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyric acid 2-amino-2- hydroxymethyl-1,3-propanediol salt (Compound no. 22) To a solution of N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyric acid (11.08 g; 0.04 mol), prepared according to the method de¬ scribed in example 1, in aqueous ethanol at 95$ (80 ml), 2-a_rd.no- 2-hydroxymethyl-l,3-propanediol (4-85 g. 0.04 mol) was added and the solution was stirred for 0 minutes at room temperature. After evaporation of the solvent, the solid residue was crystal- lized from isopropanol (l6θ ml) giving N-(3-bromo-5-isoxazolecar- bonyl)-4-aminobutyric acid 2-amino-2-hydroxymethyl-l,3-propa_ ~ .ediol salt (14.8 g; 93$ yield) with m.p. 133-135°C. 1 H-NMR (DMS0-d,): delta (ppm): 1.72 (2H, m, CH -CH -CH ); 2.14 (2H, dd, CH 2 C0); 3-25 (2H, m, 5-73 (6H, broad s, exchangeable protons); 7-35 (lH, s, -CH=); 9.39 (lH, t, CONH). In a similar way the following compound was prepared N-(3-bromo-5-isoxazolecarbonyl)-4-aminobutyric acid sodium salt (Compound no. 23) m.p. > 200°C (ethanol); 87$ yield X H-NMR (D O): delta (ppm): I.78 (2H, m, CH -CH -CH ); 2.18 (2H, dd, CH CO); 3-31 (2H, dd, N-CH ); 7-03 (lH, s, -CH=). Example 8 Pharmacodynamic evaluation: electroshock (ECS) amnesia test in rat Male rats weighing 130-150 g and fasting for at least 12 hours before the beginning of the test were used. The test consisted in two experiments with a resting interval of 24 hours between them. First experiment: training session The animals were trained to associate the passage from a light partition to a dark one, in a suitable box, with a noxious stimu¬ lation that is an electric stimulation through an electrified grid (foot-shock: 1 mA for 10 seconds). Second experiment: retention session The ability of the animals to remember was evaluated as the time utilized to repeat the passage through the partitions (retention time). — Evaluation of the retention time in control groups of animals The test was carried out first on two groups of animals as con¬ trol. A first group of rats (hereinafter indicated as control), which remembered the association between the passage and the noxious stimulation, showed a conditioned response that led to avoid the repetition of the passage. A second group of animals (hereinafter indicated as ECS control), which underwent a treatment for inducing amnesia (ECS: 90 mA, 100 Hz for 1 second) immediately after the training session, showed a decreased conditioned response and a large number of rats repeated the passage. Fixing 60 seconds as time—limit for performing the second experi¬ ment, the average time (retention time) utilized for the passage was 54 seconds for the control and 32 seconds for the ECS control. — Evaluation of the retention time in groups of treated animals Groups of 20 rats underwent the training session, the treatment for inducing amnesia and the retention session after administra¬ tion of the compounds object of the present invention and of the reference compounds, Piracetam and Aniracetam. The administration was carried out 3 minutes before the training session in case of intraperitoneal administration (i.p.) and 60 minutes before in case of administration by oral route. At least three doses each compound and each administration route were tested. The difference between the retention time of the control and the retention time of the ECS control was considered as reference (Δt) and it was fixed, as percentage, equal to 100. In the groups of animals which were treated and underwent ECS the activity against amnesia of the compounds object of the present invention was expressed as E that is as the dose of compound able to give an improvement of the retention time equal to 50% of the reference interval (Δt) in comparison with the ECS control. As example the ED values of some representatives of the com¬ pounds of formula I are reported in table 1. Table 1 Compound i.p.(umol/kg) os(μmol/kg) ∞SO E no. 1 102 290 2 160 260 3 276 868 4 202 383 8 25 80 12 171 1000 19 26l 21 254 Piracetam 329 1550 Aniracetam 353 1110

Claims: Claims 1) A compound of formula wherein n represents a number selected from 1 and 2; R represents a hydrogen atom, a halogen atom, hydroxy, a C -C, alkyl or alkoxy, R represents hydroxy, an optionally unsaturated C -C 0 alkoxy a " linear or branched C -C, alkyl, a group of formula 1 o ((CCHHJ RR α _ wwhheerreeiinn 22 mm 44 represents an integer selected from 1, 2 and 3} R represents a carboxy group, an alkoxycarbonyl group having from 1 to 6 carbon atoms in the alkoxy moiety or a group of R and R,, the same or different, represent a hydrogen atom or a " linear or branched C -C, alkyl; 1 6 i or one of R and R represents a group of formula R -CH-R wherein R has the above reported meanings and R represents a linear or branched C -C alkyl optionally substituted by phenyl, hydroxy or by a mercapto group, and the other represents a hydrogen atom; or in addition R and R ^ , together with the nitrogen atom to which they are 2 __ bonded, form a 5, 6 or 7 membered heterocycle which may further contain 1 or 2 heteroatoms selected among nitrogen, ox en and sulphur; and pharmaceutically acceptable salts thereof. 2) A compound, according to claim 1, wherein R represents a chlorine atom, a bromine atom, hydroxy, methoxy, ethoxy, methyl or ethyl. 3) A coπpound, according to one or both of the claims 1-2, wherein FL represents hydroxy. 4) A process for the preparation of the coπpounds according to one or more of the claims 1-3 comprising the condensation of a com¬ pound of formula wherein R has the meanings reported in claim 1 with 4-amino- butyric acid or a derivative thereof of formula H-(NH-CH -CH -CH -CO) -R, (II-A) 2 2 2 n 1 wherein n and R have the meanings reported in claim 1, in an inert solvent in the presence of an acceptor of halogenhydric acids. 5) A pharmaceutical composition with nootropic activity contain¬ ing a therapeutical " 1y effective amount of a compound according to one or more of the claims 1-3 optionally together with one or more excipients suitable for pharmaceutical use. 6) A method for the treatment of inpairments of cerebral function¬ ality due to ageing as well as to pathologic or traumatic rea¬ sons consisting in administering a therapeutically effective amount of a coπpound according to one or more of the claims 1-3.

Inventors: CARENZI ANGELO, CHIARINO DARIO, DELLA BELLA DAVIDE, NAPOLETANO MAURO, SALA ALBERTO, CARENZI, ANGELO, CHIARINO, DARIO, DELLA BELLA, DAVIDE, NAPOLETANO, MAURO, SALA, ALBERTO
Applicants: ZAMBON SPA, ZAMBON GROUP S.P.A.
Application year: 1988
Main classification: C07D261
IPC codes: A61K31, A61P25, C07D261, C07D413