L-PHOSPHOSERINE SALTS, A PROCESS FOR THE PREPARATION THEREOF AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM

Abstract: L-Phosphoserine derivatives, having general formula I wherein X and Y, which may be the same or different, repre­ sent the cation of a basic amino acid, such as L-Arginine or L-Lysine, show a remarkable activity on the acquirement of conditioned responses in treated animals. Said derivatives are prepared by direct salification of the acid with the desired bases.

Description: L-PHOSPHOSERINE SALTS, A PROCESS FOR THE PREPARATION THE REOF AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM The present invention relates to L-Phosphoserine amino acidic derivatives, of general formula EMI1.1 wherein X and y, which may be the same or different, represent the cation of a basic amino acid selected from Arginine and Lysine. Phosphoserine, due to its well-known activity on Central Nervous System, is used in the treatment of psychic disorders, particularly in senile involution. It has now been found that the salification products between Phosphoserine and Arginine and/or Lysine, remarkably improve the energetic balance of cerebral cell, at the same time detoxifying it from catabolic products, therefore causing, in the clinical application, an improvement in neuro-affective parameters, which may be negatively affected mainly in patients' of evolutional and involutional age. The compounds according to the present invention are obtained by reacting in aqueous solution the starting compounds, then precipitating the resulting salts by means of a non-solvent, such as a lower alcohol or a lower ketone. The following are non-limiting examples of the process for preparing the compounds of the invention. EXAMPLE 1 L-Phosphoserine salt with L-Arginine and L-Lysine (PR 157) In a 500 ml beaker containing 30 ml of water, 29.2 g of a 50% aqueous solution of L-Lysine base and 17.4 g of L-Arginine were added in succession, under stirring. Stirring was continued for about 2 hours, to complete dissolution. To the clear solution, 18.5 g of L-Phosphoserine were added progressively. To the resulting completely clear solution, 200 ml of absolute ethanol were added, under strong stirring. The reaction mixture was left to stand for 2 hours, then the obtained oily solution was separated, which was slowly dropped into other 200 ml of absolute ethanol, under strong stirring. After stirring for 4 hours, the mixture was filtered, the residue was washed with some ethanol and dried under vacuum overnight. 45 Grams were obtained (yield: 89% on theoric). M.p. 112-1150C. EIemental Analysis Calc.% C = 35.64; H = 7.17; 0 = 31.65 Found % C = 35.94; H = 6.84; 0 = 31.32. EXAMPLE 2 L-Phosphoserine salt with L-Arginine 1:2 (PR 156) In a 500 ml beaker containing 50 ml of water, 18.5 g of L-Phosphoserine, then 34.8 g of L-arginine, were added, under stirring. Stirring was continued for about 3 hours, to complete dissolution. The resulting solution was diluted with 200 ml of acetone, to obtain an oily solution, which was decanted and slowly dropped in other 200 ml of acetone, under strong stirring. Stirring was continued for 4 hours, then the mixture was filtered, the residue was washed with some ethyl acetate and dried under vacuum overnight. 52 Grams of title compound were obtained (yield 97.4% on theoric), melting at 80-85"C. Analogously to the procedure described in Example 2, the L-Lysine double salt was prepared. The elementary analysis is consistent with the structure of the compound. Pharmacological properties The compounds of the present invention were pharmacologically tested, in order to investigate any activity on acquirement processes, memory or depression. Male young and elderly Wistar rats, weighing respectively 30 and 580 g, were used. The test compounds, dissolved in physiological saline, were administered orally to young and elderly animals, at doses of 5, 10 and 20 mg/kg. Phosphoserine dissolved in physiological saline was administered at equimolecular doses (1.8, 3.6 and 7.2 mg/kg). The control animals were administered with the sole vehicle. The administrations were carried out once a day during 20 days, the last administration being effected 1 hour before the tests. The following behavioral tests were carried out: 1) Shuttle box test: to evaluate the acquirement of conditioned avoidance responses (CARs). The results reported in Table 1 show that PR 157 caused an increase in the number of conditioned responses as well as in the percentage of learners, both in young and elderly animals. Phosphoserine caused similar effects on the elderly animals only. PR 156, which was tested only on young rats, proved also to be effective. The activity was dose-related, and higher in the elderly animals treated with PR 157 than in those treated with Phosphoserine at equimolecular doses. TABLE 1 EMI4.1 <tb> TEST <SEP> GROUPS <SEP> CARs* <SEP> LEARNERS <SEP> (%) <tb> Young <SEP> rats <tb> Vehicle <SEP> (24) <SEP> 16.0 <SEP> + <SEP> 0.3 <SEP> 50.0 <tb> PR <SEP> 156 <tb> 5 <SEP> mg/kg <SEP> (6) <SEP> 23.4 <SEP> + <SEP> 1.9** <SEP> 100.0 <tb> 10 <SEP> mg/kg <SEP> (4) <SEP> 24.0 <SEP> + <SEP> 1.8** <SEP> 100.0 <tb> PR <SEP> 157 <tb> 5 <SEP> mg/kg <SEP> (12) <SEP> 19.4 <SEP> + <SEP> 0.4* <SEP> 50.0 <tb> 10 <SEP> mg/kg <SEP> (12) <SEP> 22.2 <SEP> + <SEP> 1.4* <SEP> 75.0 <SEP> <tb> 20 <SEP> mg/kg <SEP> (12) <SEP> 24.4 <SEP> + <SEP> 1.4* <SEP> lOO.0** <SEP> <tb> Phosphoser <SEP> ine <SEP> <tb> 1.8 <SEP> mg/kg <SEP> (8) <SEP> 16.1 <SEP> + <SEP> 0.5 <SEP> 50.0 <tb> 3.6 <SEP> mg/kg <SEP> (8) <SEP> 17.6 <SEP> + <SEP> 0.5 <SEP> 50.0 <tb> 7.2 <SEP> mg/kg <SEP> (8) <SEP> 17.2 <SEP> + <SEP> 0.6 <SEP> 75.0 <tb> Elderly <SEP> rats <tb> Vehicle <SEP> (22) <SEP> 3.7 <SEP> + <SEP> 0.2 <SEP> 0.0 <tb> PR <SEP> 157 <tb> 5 <SEP> mg/kg <SEP> (12) <SEP> 7.9 <SEP> + <SEP> 0.3* <SEP> 25.0 <tb> 10 <SEP> mg/kg <SEP> (12) <SEP> 11.5 <SEP> + <SEP> 0.3* <SEP> 50.O** <SEP> <tb> 20 <SEP> mg/kg <SEP> (12) <SEP> 18.6 <SEP> + <SEP> 0.4* <SEP> 50.0 <SEP> <tb> Phosphoser <SEP> ine <SEP> <tb> 1.8 <SEP> mg/kg <SEP> (8) <SEP> 5.9 <SEP> + <SEP> 0.4* <SEP> 25.0 <tb> 3.6 <SEP> mg/kg <SEP> (8) <SEP> 6.2 <SEP> + <SEP> 0.5* <SEP> 37.5" <SEP> <tb> 7.2 <SEP> mg/kg <SEP> (8) <SEP> 9.2 <SEP> + <SEP> 0.6* <SEP> 37.5 <SEP> <tb> * Mean values + ES ** Significant difference in comparison with controls (p < 0.05, Dunnett's test). Number of animals for each group is between brackets. 2. Dark box test: to further evaluate the effect on memory capability of the animals. The results are reported in Table 2. PR 157 caused a dose-related improvement in the acquirement of avoidance responses both in young and elderly animals, whilst Phosphoserine proved to be effective only on the elderly animals. Moreover, PR 157 had an higher activity than that of Phosphoserine. TABLE 2 EMI5.1 <tb> TEST <SEP> GROUPS <SEP> LATENCY <SEP> (seconds) <tb> <SEP> Young <SEP> rats <SEP> Elderly <SEP> rats <tb> Vehicle <SEP> (24) <SEP> 80.0 <SEP> (24) <SEP> 20.0 <tb> PR <SEP> 157 <SEP> 5 <SEP> mg/kg <SEP> (12) <SEP> 112.3* <SEP> (12) <SEP> 46.0* <tb> <SEP> 10 <SEP> mg/kg <SEP> (12) <SEP> 161.5* <SEP> (12) <SEP> 79.4* <SEP> <tb> <SEP> 20 <SEP> mg/kg <SEP> (12) <SEP> 265.1* <SEP> (12) <SEP> 82.0* <tb> Phosphoserine <SEP> 1.8 <SEP> mg/kg <SEP> (12) <SEP> 97.5* <SEP> (12) <SEP> 34.8 <tb> <SEP> 3.6 <SEP> mg/kg <SEP> (12) <SEP> 110.2* <SEP> (12) <SEP> 50.7* <tb> <SEP> 7.2 <SEP> mg/kg <SEP> (12) <SEP> 186.0* <SEP> (12) <SEP> 65.0* <tb> The values are averages. * Significant difference in comparison with controls (P < 0.05, Mann-Whitney test). The above tests prove that the compounds of the invention, particularly PR 157, have a marked dose-related activity on acquirement and memory processes, as well as on neuro-affective parameters, The number of animals for each group is between brackets. 3. Despair test: to estimate the antidepressant activity on the elderly rats. The results are illustrated in Table 3. PR 157 and Phosphoserine showed a dose-related reduction in the stillness of the elderly rats. At equimolecular doses, the activities of PR 157 and Phosphoserine were substantially superimposable. TABLE 3 EMI6.1 <tb> TEST <SEP> GROUPS <SEP> DURATION <SEP> OF <SEP> STILLNESS <tb> Vehicle <SEP> (24) <SEP> 320.5 <SEP> + <SEP> 42.6 <tb> PR <SEP> 157 <SEP> 5 <SEP> mg/kg <SEP> (12) <SEP> 280.2 <SEP> + <SEP> 20.3* <tb> <SEP> 10 <SEP> mg/kg <SEP> (12) <SEP> 263.1 <SEP> + <SEP> 33.4* <tb> <SEP> 20 <SEP> mg/kg <SEP> (12) <SEP> 236.7 <SEP> + <SEP> 28.9* <tb> Phosphoserine <SEP> 1.8 <SEP> mg/kg <SEP> (12) <SEP> 301.0 <SEP> + <SEP> 31.4* <tb> <SEP> 3.6 <SEP> mg/kg <SEP> (12) <SEP> 291.7 <SEP> + <SEP> 32.7* <tb> <SEP> 7.2 <SEP> mg/kg <SEP> (12) <SEP> 265.8 <SEP> + <SEP> 24.7* <tb> Mean values + SEM * Significant difference in comparison with controls (P < 0.05, Dunnett's test). The number of animals for each group is between brackets. The above tests prove that the compounds of the invention, particularly PR 157, have a marked dose-related activity on acquirement and memory processes, as well as on neuro-affective parameters, both in young and elderly animals, said activity being higher than that of Phosphoserine. Further, it should be pointed out that Phosphoserine is active substantially only on elderly animals, showing poor effects on the young ones. The compounds of the invention may be used in human therapy, for the treatment of neuro-affective deficiencies of both involutional and evolutional age, acquirement and memory deficiencies of evolutional age, intellection deficiencies and mental deterioration of senile involution. The present invention also relates to all the indu strial aspects connected with the use of compounds of general formula I as therapeutic agents. Therefore, a primary object of the invention is provided by pharmaceutical compositions containing, as the active ingredient, pre-set therapeutically effective amounts of at least one compound of formula I, optionally in admixture with conventional excipients. Examples of said pharmaceutical compositions comprise tablets, dragees, capsules, syrups, sachets, vials for oral and parenteral administration. The preferred dosage will vary, depending on conditions, weight and age of the patients, from about 150 to about 900 mg/day. Non-limiting examples of pharmaceutical compositions according to the invention comprise: 150 and 300 mg tablets; 75 and 150 mg pediatric oral vials (in 5 ml); 150 and 300 mg adult oral vials (in 10 ml); 150 and 300 mg sachets; 100 and 150 mg i.m. vials.

Claims: CLAIMS for the contracting states: BE, CH, DE, FR, GB, IT, LI, LU, NL, SE 1. L-Phosphoserine amino acidic derivatives, of general formula I EMI8.1 wherein X and Y, which may be the same or different, represent the cation of a basic amino acid, selected from L-Arginine and L-Lysine. 2. A compound as claimed in claim 1, wherein X = L-Arginine and Y = L-Lysine. 3. A compound as claimed in claim 1, wherein X = Y = L-Arginine. 4. A compound as claimed in claim 1, wherein X = Y = L-Lysine. 5. A process for preparing any one of the compounds as claimed in claims 1-4, wherein L-Phosphoserine is reacted with L-Arginine and/or L-Lysine, in an aqueous solution, in appropriate molar ratios, and the resulting salt is precipitated by means of a non-solvent. 6. A process as claimed in claim 5, wherein the nonsolvent is a lower alcohol, particularly ethanol. 7. A process as claimed in claim 5, wherein the nonsolvent is a lower ketone, particularly acetone. 8. Pharmaceutical compositions for the treatment of neuro-affective deficiencies of both involutional and evo lutional age, acquirement and memory deficiencies of evolutional age, intellection deficiencies and mental deterioration of senile involution, containing as the active ingredient at least one compound as claimed in claims 1 to 4. 9. Pharmaceutical compositions as claimed in claim 8, for oral and parenteral administration, in form of tablets, capsules, syrups, sachets, oral vials and intramuscular injection vials. CLAIMS for AT: 1. A process for the preparation of L-Phosphoserine amino acidic derivatives, of general formula I EMI10.1 wherein X and Y, which may be the same or different, represent the cation of a basic amino acid, selected from L-Arginine and L-Lysine, which process consists in reacting L-Phosphoserine with L-Arginine and/or L-Lysine, in an aqueous solution, in appropriate molar ratios, and precipitating the resulting salt by means of a non-solvent. 2. A process as claimed in claim 1, wherein the nonsolvent is a lower alcohol, particularly ethanol. 3. A process as claimed in claim 1, wherein the nonsolvent is a lower ketone, particularly acetone.

Inventors: VOLTERRA, BRUNO
Applicants: LABORATORI PROPHIN S.P.A.
Application year: 1986
Main classification: C07F9
IPC codes: C07F9