(2,5-DIOXOPYRROLIDIN-1-YL)(PHENYL)-ACETAMIDE DERIVATIVES AND THEIR USE IN THE TREATMENT OF NEUROLOGICAL DISEASES

Abstract

The first object of the invention is the compound of general formula (I) or pharmaceutically acceptable salts thereof. A second object of the invention is the use of compounds described by general formula (I) as active ingredient in pharmaceutical compositions for the treatment of epileptic seizures or neuropathic pain or migraine.

##STR00001##

Claims

1.-14. (canceled)

15. A compound of the general formula (I) or pharmaceutically acceptable salts thereof, ##STR00005## wherein: X is N or C, k is a number equal to 0 or 1, A is a substituent selected from the group comprising of: phenyl substituent; a phenyl substituent substituted with one or two or three or four side substituents selected from the group comprising of: halogen atoms, —SCF.sub.3, —CF.sub.3, —CHF.sub.2, —CN, —OCF.sub.3, —NO.sub.2, —OCH.sub.3, —OC.sub.2H.sub.5, an alkyl moiety with the number of carbon atoms in the carbon backbone from 1 to 4, wherein the alkyl moiety has a straight or branched chain; a phenyl substituent substituted with at least one aromatic or heteroaromatic substituent; a benzhydryl substituent; a 1-naphthyl or 2-naphthyl substituent; a benzothiophenyl substituent selected from the group comprising of: 2-benzothiophenyl, 3-benzothiophenyl, 4-benzothiophenyl or 5-benzothiophenyl substituents, preferably 5-benzothiophenyl substituent; a benzisoxazole substituent selected from the group comprising of: 3-benzisoxazole, 4-benzisoxazole, 5-benzisoxazole, 6-benzisoxazole, 7-benzisoxazole substituents, preferably 5-benzisoxazole substituent; an alkyl moiety with the number of carbon atoms in the carbon backbone from 1 to 4, wherein the alkyl moiety has a straight or branched or cyclic chain, preferably the alkyl moiety is substituted with at least one halogen atom; B is: phenyl substituent; a phenyl substituent substituted with one or two side substituents selected from the group comprising of: halogen atoms, —SCF.sub.3, —CF.sub.3, —CHF.sub.2, —CN, —OCF.sub.3, —NO.sub.2, —OCH.sub.3, —OC.sub.2H.sub.5, an alkyl moiety with the number of carbon atoms in the carbon backbone from 1 to 4, wherein the alkyl moiety has a straight or branched chain; D is a substituent selected from the group comprising of: H, amino (—NH.sub.2), amino group substituted with one or two aliphatic substituents (including in particular —CH.sub.3 and/or —C.sub.2H.sub.5) or an amino group which is part of a heterocyclic ring.

16. The compound according to claim 15, characterized in that it is a compound of the general formula (II) or a pharmaceutically acceptable salt thereof ##STR00006## wherein: X is N or C, k is a number equal to 0 or 1, A is a substituent selected from the group comprising of: phenyl substituent; a phenyl substituent substituted with one or two or three or four side substituents selected from the group comprising of: halogen atoms, —SCF.sub.3, —CF.sub.3, —CHF.sub.2, —CN, —OCF.sub.3, —NO.sub.2, —OCH.sub.3, —OC.sub.2H.sub.5, an alkyl moiety with the number of carbon atoms in the carbon backbone from 1 to 4, wherein the alkyl moiety has a straight or branched chain; a phenyl substituent substituted with at least one aromatic or heteroaromatic substituent; a benzhydryl substituent; a 1-naphthyl or 2-naphthyl substituent; a benzothiophenyl substituent selected from the group comprising of: 2-benzothiophenyl, 3-benzothiophenyl, 4-benzothiophenyl or 5-benzothiophenyl substituents, preferably 5-benzothiophenyl substituent; a benzisoxazole substituent selected from the group comprising of: 3-benzisoxazole, 4-benzisoxazole, 5-benzisoxazole, 6-benzisoxazole, 7-benzisoxazole substituents, preferably 5-benzisoxazole substituent; an alkyl moiety with the number of carbon atoms in the carbon backbone from 1 to 4, wherein the alkyl moiety has a straight or branched or cyclic chain, preferably the alkyl moiety is substituted with at least one halogen atom; B is: phenyl substituent; a phenyl substituent substituted with one or two side substituents selected from the group comprising of: halogen atoms, —SCF.sub.3, —CF.sub.3, —CHF.sub.2, —CN, —OCF.sub.3, —NO.sub.2, —OCH.sub.3, —OC.sub.2H.sub.5, an alkyl moiety with the number of carbon atoms in the carbon backbone from 1 to 4, wherein the alkyl moiety has a straight or branched chain.

17. The compound according to claim 15, characterized in that the halogen atom is a fluorine or chlorine atom.

18. The compound according to claim 15, characterized in that the alkyl moiety in the carbon backbone contains from 1 to 4 carbon atoms, wherein the alkyl moiety has a straight or branched chain and is selected from the group comprising of: methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl.

19. The compound according to claim 15, characterized in that k=0.

20. The compound according to claim 15, characterized in that X is nitrogen.

21. The compound according to claim 15, characterized in that the substituent A is selected from the group comprising of: 5-benzothiophenyl, 2-naphthyl, 5-benzisoxazolyl substituents.

22. The compound according to claim 15, characterized in that the substituent A is selected from the group comprising of: phenyl, phenyl substituted with at least one chlorine atom or —CF.sub.3, —CHF.sub.2, —OCF.sub.3, —CH.sub.3, —SCF.sub.3 or phenyl.

23. The compound according to claim 15, characterized in that the substituent B is selected from the group comprising of phenyl or phenyl substituted with one or two halogen atoms.

24. The compound according to claim 15, characterized in that it is selected from the group comprising of: 1-(2-Oxo-1-phenyl-2-(4-phenylpiperazin-1-yl)ethyl) pyrrolidine-2,5-dione 1-(2-(4-(3-Chlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione 1-(2-(4-(3,5-Dichlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(m-tolyl)piperazin-1-yl) ethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(4-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-(4-(3,5-Bis(trifluoromethyl) phenyl) piperazin-1-yl)-2-oxo-1-phenylethyl) pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(3-(difluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethoxy) phenyl) piperazin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(4-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl(sulfanyl)phenyl) piperazin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-(4-([1,1′-Biphenyl]-3-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione 1-(1-(4-Fluorophenyl)-2-oxo-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-(4-(Naphth-2-yl) piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione 1-(2-(4-(Benzo[b]thiophen-5-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione 1-(2-(4-(1,2-Benzoxazol-5-il)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione 1-(2-(4-(3-Chlorophenyl)piperidin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperidin-1-yl)ethyl)pyrrolidine-2,5-dione 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethoxy) phenyl) piperidin-1-yl)ethyl)pyrrolidine-2,5-dione.

25. The compound according to claim 15, characterized in that it is a (R) enantiomer, preferably selected from the following compounds: (R)-1-(2-(4-(3-chlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione, (R)-1-(2-(4-(3,5-dichlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione, (R)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione, (R)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione, (R)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl(sulfanyl)phenyl) piperazin-1-yl)ethyl)pyrrolidine-2,5-dione.

26. The compound according to claim 15, characterized in that it is a water-soluble salt, especially a hydrochloride salt, preferably selected from the following compounds: 3-(Methylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl) phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride, 3-(Dimethylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride, 3-(Diethylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifloromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride.

27. A method for treating or preventing epileptic seizures, neuropathic pain or migraine comprising administering the compound in claim 15.

Description

[0086] Embodiments of the invention are illustrated in Figures, where it is shown: FIG. 1—general formula of compound (1) and (II); FIG. 2A—the synthesis of derivatives according to formula (I); FIG. 2B—the synthesis of derivatives according to formula (II); FIG. 3—analgesic activity of compound 6 and valproic acid (VPA) in phase I and 11 pain of the formalin test, where the results are presented as time of paw licking in the first phase of the test (0-5 minutes after formalin injection) and in phase II (15-30 minutes after formalin injection), the values represent mean±SEM for a group of 8-10 animals; statistically significant difference in comparison to the control (vehicle—Tween) group, statistical analysis—one-way ANOVA analysis of variance, Dunnett's post hoc test: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. C—control group; VPA—valproic acid; FIG. 4—analgesic activity of compound 6 and valproic acid (VPA) in the capsaicin assay, where the results are shown as paw licking time 0-5 minutes after capsaicin injection, the values represent mean±SEM; statistically significant difference in comparison to the control (vehicle—Tween) group, statistical analysis—one-way ANOVA analysis of variance, Dunnett's post hoc test: *p<0.05, **p<0.01, ****p<0.0001. C—control group; VPA—valproic acid; FIG. 5—analgesic activity of compound 6 and valproic acid (VPA) in the oxaliplatin-induced peripheral neuropathy, where: A—the effect of compound 6 on mechanical allodynia in the von Frey test. B—the effect of valproic acid (VPA) on mechanical allodynia in the von Frey test. C—the effect of compound 6 on thermal allodynia in the Cold Plate test, the results are presented as the mean value of pressure force causing paw lift (von Frey test) or latency time to the occurrence of the nociceptive cold-plate reaction±SEM for in group of 8-10 animals; statistically significant difference compared to the control group (mice after administration of OXPT and before administration of test compounds, statistical analysis—one-way ANOVA analysis of variance, Dunnett's post hoc test: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. Veh—vehicle (1% Tween 80); FIG. 6—the effect of compound 6 (at a concentration of 100 μM) on fast, potential-dependent sodium currents, where: A—example traces of maximal voltage-gated sodium currents in control, in the presence of compound 6 and after wash-out of compound 6; B—averaged normalized current amplitudes in control, in the presence of the compound 6 (*p<0.001, ANOVA with Tukey's test), and after wash-out of compound 6. I/Imax (on the vertical axis) means that the currents were normalized to control values; FIG. 7—UPLC analysis of the metabolism of compound 6 after incubation with HMLs; FIG. 8—the effect of verapamil, Na.sub.3VO.sub.4 and compound 6 on baseline Pgp activity, statistical significance were calculated by one-way ANOVA variance analysis and the Bonferroni method (**p<0.01, ***p<0.001, compounds tested in triplicate); FIG. 9 A—effect of the reference inhibitor ketoconazole (KE) and compound 6 on CYP3A4 activity; B—the effect of the reference inhibitor quinidine (QD) and 6 on CYP2D6 activity. Statistical significance was calculated by one-way ANOVA variance analysis and the Bonferroni method (***p<0.001); FIG. 10—effect of reference cytostatic doxorubicin (DX), mitochondrial toxin CCCP (carbonyl cyanide m-chlorophenylhydrazone) and compound 6 on cell viability of the HepG2 line after 72 h incubation. Statistical significance was calculated by one-way ANOVA variance analysis and the Bonferroni method (*p<0.05, ***p<0.001, compounds tested in four replications); FIG. 11—ATP level in HepG2 cell line after 3 hours incubation. Doxorubicin (DX), CCCP (carbonyl cyanide m-chlorophenylhydrazone). Statistical significance was calculated by one-way ANOVA variance analysis and the Bonferroni method (***p<0.001, compounds tested in four replications); FIG. 12—general scheme for the synthesis of enantiomers of compounds according to formula (II); FIG. 13—UPLC analysis of (R)-6 enantiomer metabolism after incubation with HMLs; FIG. 14—general scheme for the synthesis of water-soluble salts of compounds according to formula (I).

[0087] Analytical Methods:

[0088] Proton magnetic resonance (.sup.1H NMR) and carbon nuclear magnetic resonance (.sup.13C NMR) spectra were recorded using a Mercury-300 “Varian” spectrometer (Varian Inc., Palo Alto, Calif., USA) at 300 MHz and 75 MHz, respectively, or JEOL-500 spectrometer (JEOL USA, Inc. MA, USA), operating at 500 MHz and 126 MHz, respectively. Chemical shifts are given in δ (ppm) values relative to TMS 6=0 (1H) as an internal standard. J values are expressed in hertz (Hz). Deuterated chloroform (CDCl.sub.3) or deuterated dimethyl sulfoxide (DMSO-D.sub.6) was used as the solvent. The following signal abbreviations have been used in the spectra description: s (singlet), br s (broad singlet), d (doublet), dd (doublet of doublets), ddd (doublet of doublet of doublets), t (triplet), td (triplet of doublets), q (quartet), m (multiplet).

[0089] The UPLC/MS analysis system consisted of a Waters ACQUITY® UPLC® apparatus (Waters Corporation, Milford, Mass., USA) coupled with a Waters TQD mass spectrometer operating in electrospray ionization (ESI) mode. Chromatographic separations were carried out using the Acquity UPLC BEH C18, 1.7 μm (2.1×100 mm) column with the VanGuard Acquity UPLC BEH C18, 1.7 μm (2.1×5 mm) (Waters, Milford, Conn., USA). The column was maintained at 40° C. and eluted with a gradient of 95% to 0% of eluent A over 10 min, with a flow rate of 0.3 mL/min. Eluent A: water/formic acid (0.1%, v/v); eluent B: acetonitrile/formic acid (0.1%, v/v). Chromatograms were recorded using a Waters eλ PDA detector. Spectra were analyzed in the 200-700 nm range with a resolution of 1.2 nm and a sampling rate of 20 points/s. The UPLC retention times (t.sub.R) are given in minutes. Thin layer chromatography (TLC) was performed on aluminum sheets coated with silica gel 60 F.sub.254 (Macherey-Nagel, Düren, Germany), using developing solvent systems with the following composition: DCM:MeOH (9:0.2; v/v), DCM:MeOH (9:0.3; v/v), DCM:MeOH (9:0.5; v/v), DCM:MeOH (9:1, v/v). Spot detection—UV light (λ=254 nm). Melting points (m.p.) were determined using open capillaries in a BQchi 353 apparatus (BQchi Labortechnik, Flawil, Switzerland). Enantiomeric purity was determined using a chiral HPLC technique on a Shimadzu Prominence and LC-2030C SD Plus apparatus (Shimadzu Corporation, Kyoto, Japan) equipped with an Amylose-C (250×4.6 mm) chiral column. The analysis was performed under the following conditions: column temperature: 20° C., mixture of eluents: hexane/i-PrOH=80/20 (v/v), flow: 1 mL/min, detection at the wavelength λ=206 nm. Enantiomeric purity is expressed in %.

[0090] The preparation of compounds of the invention is illustrated in the following examples. The syntheses presented in the examples below were not optimized in terms of yield, amount of reagents used or the final form of obtained compounds.

Abbreviations Used

[0091] AcOEt—ethyl acetate
CDI—carbonyldiimidazole
DCC—N,N′-dicyclohexylcarbodiimide
DCM—dichloromethane
DMF—dimethylformamide
Et.sub.2O—diethyl ether
HCl—hydrochloric acid
HMDS—hexamethyldisilazane
MeOH—methanol
NaCl—sodium chloride
Na.sub.2SO.sub.4— sodium sulfate
ZnCl.sub.2— zinc chloride

Example 1. Synthesis, Physicochemical and Spectral Data of Intermediates (IV and III According to the Scheme in FIG. 2B)

Intermediate IV: 4-((Carboxy(phenyl)methyl)amino)-4-oxobutanoic Acid

[0092] Succinic anhydride (3.0 g, 30 mmol, 1 eq) was dissolved in 15 mL of glacial acetic acid, followed by the addition of an equimolar amount of DL-phenylglycine (4.53 g). The mixture was heated at 70° C. with stirring for 12 hours. After this time, acetic acid was distilled off to dryness. Intermediate IV was obtained as a solid after washing with Et.sub.2O.

[0093] White solid. Yield: 87% (6.55 g); m.p. 199.4-200.6° C.; TLC: R.sub.f=0.25 (DCM:MeOH (9:1; v/v)); C.sub.12H.sub.13NO.sub.5 (251.24), Monoisotopic mass: 251.08. UPLC (100% purity): t.sub.R=2.77 min. (M+H)+252.1.

Intermediate III: 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic Acid

[0094] ZnCl.sub.2 (2.73 g, 20 mmol, 1 eq) was added to a suspension of 4-((carboxy(phenyl)methyl)amino)-4-oxobutanoic acid (5.0 g, 20 mmol, 1 eq) (IV) in dry benzene (100 mL) and heated to 80° C. Then a solution of HMDS (4.84 g, 6.25 mL, 30 mmol, 1.5 eq) in dry benzene (15 mL) was added dropwise over 30 minutes. The reaction was continued with stirring at reflux for about 24 hours and next concentrated under reduced pressure. After renoval off the solvent, the oily residue was dissolved in DCM and extracted with 0.1 M HCl (3×50 mL), water (3×50 mL) and saturated NaCl solution (3×50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and then evaporated to dryness. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid was obtained as a solid substance after washing with Et.sub.2O. Alternatively, 1,4-dioxane can be used instead of benzene in the above procedure.

[0095] White solid. Yield: 90% (4.20 g); m.p. 195.5-198.2° C.; TLC: R.sub.f=0.45 (DCM:MeOH (9:1; v/v)); C.sub.12H.sub.11NO.sub.4 (233.22), Monoisotopic mass: 233.07. UPLC (100% purity): t.sub.R=3.41 min. (M+H)+234.1. .sup.1H NMR (300 MHz, DMSO-D.sub.6) δ 2.73 (s, 4H), 5.76 (s, 1H), 7.26-7.35 (m, 3H), 7.36-7.45 (m, 2H), 13.22 (br s, 1H).

Example 2. 1-(2-Oxo-1-phenyl-2-(4-phenylpiperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0096] Carbonyldiimidazole (1.17 g, 7.2 mmol, 1.2 eq) was dissolved in 5 mL of dry DMF and then added to a solution of 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) dissolved in 10 mL of anhydrous DMF. After stirring for 0.5 hour, a solution of 1-phenylpiperazine (0.97 g, 6 mmol, 1 eq) in 5 mL of anhydrous DMF was added dropwise. The reaction was continued with stirring at room temperature for 24 hours. After this time, DMF was distilled off under reduced pressure. The crude product was purified by column chromatography using mixture of DCM:MeOH (9:0.3; v/v) as solvent system. The compound was obtained as a solid after washing with Et.sub.2O.

[0097] White solid. Yield: 84% (1.90 g); m.p. 156.7-157.4° C.; TLC: R.sub.f=0.35 (DCM:MeOH (9:0.3; v/v)); C.sub.22H.sub.23N.sub.3O.sub.3 (377.44), Monoisotope mass: 377.17. UPLC (100% purity): t.sub.R=5.88 min. (M+H).sup.+378.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.58-2.81 (m, 5H), 2.95-3.15 (m, 2H), 3.17-3.42 (m, 3H), 3.63-3.76 (m, 1H), 3.92-4.05 (m, 1H), 6.12 (s, 1H), 6.80-6.91 (m, 3H), 7.19-7.28 (m, 2H) 7.29-7.47 (m, 5H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.1, 42.4, 45.8, 48.9, 49.2, 56.8, 116.5, 116.6, 120.6, 128.6, 128.6, 128.9, 129.1, 129.2, 129.8, 129.9, 133.0, 150.7, 165.0, 176.3.

Example 3. 1-(2-(4-(3-Chlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0098] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-(3-chlorophenyl) piperazine (1.40 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.2; v/v) eluent system.

[0099] White solid. Yield: 81% (2.00 g); m.p. 128.1-129° C.; TLC: R.sub.f=0.51 (DCM:MeOH (9:0.2; v/v)); C.sub.22H.sub.22ClN.sub.3O.sub.3 (411.89), Monoisotopic mass: 411.13. UPLC (100% purity): t.sub.R=6.69 min, (M+H).sup.+412.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.58-2.73 (m, 4H), 3.00 (br s, 1H), 3.27-3.53 (m, 3H), 3.54-3.86 (m, 2H), 4.17 (br s, 2H), 6.02 (s, 1H), 7.27-7.40 (m, 7H), 7.51-7.63 (m, 2H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.0, 40.0, 43.3, 53.3, 53.7, 56.5, 118.9, 120.8, 128.9, 129.1, 129.3, 129.6, 131.4, 132.1, 135.9, 143.8, 165.5, 176.7.

Example 4. 1-(2-(4-(3,5-Dichlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0100] The compound was prepared according to procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-(3,5-dichlorophenyl)piperazine (1.20 g, 6 mmol, 1 eq). The crude product was purified by column chromatography using DCM:MeOH (9:0.3; v/v) eluent system.

[0101] White solid. Yield: 77% (2.06 g); m.p. 163.8-165.2° C.; TLC: R.sub.f=0.42 (DCM:MeOH (9:0.2; v/v)); C.sub.22H.sub.21Cl.sub.2N.sub.3O.sub.3 (446.33), Monoisotopic mass: 446.10. UPLC (99% purity): t.sub.R=7.59 min, (M+H)+446.1. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.63-2.78 (m, 5H), 2.98-3.13 (m, 2H), 3.20-3.36 (m, 3H), 3.59-3.68 (m, 1H), 3.97-4.00 (m, 1H), 6.09 (s, 1H), 6.64 (d, J=1.7 Hz, 2H), 6.80 (t, J=1.7 Hz, 1H), 7.33-7.38 (m, 3H), 7.42 (d, J=6.7 Hz, 2H). .sup.13C NMR (126 MHz, CDCl.sub.3) δ 28.1, 42.2, 45.4, 48.0, 48.2, 56.9, 114.4, 119.8, 128.8, 129.1, 129.9, 132.9, 135.6, 152.1, 165.2, 176.4.

Example 5. 1-(2-Oxo-1-phenyl-2-(4-(m-tolyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0102] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-(3-methylphenyl)piperazine (1.18 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.3; v/v) eluent system.

[0103] White solid. Yield: 86% (2.02 g); m.p. 188.7-192.1° C.; TLC: R.sub.f=0.45 (DCM:MeOH (9:0.3; v/v)); C.sub.23H.sub.25N.sub.3O.sub.3 (391.47), Monoisotopic mass: 391.19. UPLC (98.9% purity): t.sub.R=6.35 min, (M+H)+392.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.36 (s, 3H), 2.57-2.78 (m, 5H), 2.91-3.54 (m, 3H), 3.63-4.55 (m, 4H), 6.06 (s, 1H), 7.22 (d, 1H, J=7.5 Hz), 7.27-7.62 (m, 8H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 21.4, 28.1, 39.7, 43.0, 54.1, 54.6, 56.5, 117.9, 121.7, 128.9, 129.3, 129.7, 130.2, 130.8, 132.3, 141.0, 141.8, 165.4, 176.3.

Example 6. 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0104] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-[3-(trifluoromethyl)phenyl]piperazine (1.38 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.2; v/v) eluent system.

[0105] White solid. Yield: 82% (2.19 g); m.p. 150.3-151.4° C.; TLC: R.sub.f=0.34 (DCM:MeOH (9:0.2; v/v)); C.sub.23H.sub.22F3N.sub.3O.sub.3 (445.44), Monoisotopic mass: 445.16. UPLC (100% purity): t.sub.R=6.94 min, (M+H)+446.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.60-2.86 (m, 5H), 3.00-3.20 (m, 2H), 3.23-3.44 (m, 3H), 3.62-3.76 (m, 1H), 3.93-4.06 (m, 1H), 6.12 (s, 1H), 6.94-7.04 (m, 2H), 7.09 (d, 1H, J=7.7 Hz), 7.28-7.51 (m, 6H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.0, 42.2, 45.6, 48.4, 48.6, 56.8, 112.7 (q, J=4.6 Hz), 116.7 (q, J=4.6 Hz), 119.2, 123.4 (q, J=271.8 Hz), 128.7, 128.9, 129.7, 129.8, 131.5 (q, J=31.8 Hz), 132.9, 150.8, 165.1, 176.3.

Example 7. 1-(2-Oxo-1-phenyl-2-(4-(4-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0106] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-[4-(trifluoromethyl)phenyl]piperazine (1.38 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.3; v/v) eluent system.

[0107] White solid. Yield: 62% (1.66 g); m.p. 173.2-174.3° C.; TLC: R.sub.f=0.49 (DCM:MeOH (9:0.3; v/v)); C.sub.23H.sub.22F3N.sub.3O.sub.3 (445.44), Monoisotopic mass: 445.16. UPLC (100% purity): t.sub.R=6.89 min, (M+H)+446.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.61-2.85 (m, 5H), 3.04-3.43 (m, 5H), 3.63-3.77 (m, 1H), 3.91-4.05 (m, 1H), 6.12 (s, 1H), 6.83 (d, 2H, J=8.6 Hz), 7.30-7.40 (m, 3H), 7.40-7.50 (m, 4H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.0, 42.1, 45.4, 47.6, 47.9, 56.8, 115.0, 124.5 (q, J=270.6 Hz), 126.5 (q, J=4.6 Hz), 128.7, 128.8, 128.9, 129.8, 132.8, 152.7, 165.1, 176.3.

Example 8. 1-(2-(4-(3,5-Bis(trifluoromethyl)phenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0108] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-[3,5-bis(trifluoromethyl)phenyl]piperazine (1.18 g, 6 mmol, 1 eq) were used as starting materials.

[0109] The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0110] White solid. Yield: 69% (2.12 g); m.p. 228.1-229.4° C.; TLC: R.sub.f=0.47 (DCM:MeOH (9:0.5; v/v)); C.sub.24H.sub.21F6N.sub.3O.sub.3 (513.44), Monoisotopic mass: 513.13. UPLC (100% purity): t.sub.R=6.58 min, (M+H)+514.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.52-2.75 (m, 4H), 2.82-3.07 (m, 4H), 3.12-3.86 (m, 4H), 6.11 (s, 1H), 6.97-7.05 (m, 3H), 7.22-7.61 (m, 5H).

Example 9. 1-(2-Oxo-1-phenyl-2-(4-(3-(difluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0111] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-(3-difluoromethylphenyl)piperazine (1.27 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM: MeOH (9:0.2; v/v) eluent system.

[0112] White solid. Yield: 83% (2.13 g); m.p. 156.4-157.6° C.; TLC: R.sub.f=0.55 (DCM:MeOH (9:0.2; v/v)); C.sub.23H.sub.23F2N.sub.3O.sub.3 (427.45), Monoisotopic mass: 427.17. UPLC (100% purity): t.sub.R=6.36 min, (M+H).sup.+428.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.58-2.78 (m, 5H), 3.02-3.18 (m, 2H), 3.24-3.46 (m, 3H), 3.62-4.08 (m, 2H), 6.12 (s, 1H), 6.44-7.62 (m, 1H), 6.94-7.04 (m, 2H), 7.28-7.51 (m, 7H).

Example 10. 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0113] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-[3-(trifluoromethoxy)phenyl]piperazine (1.48 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.3; v/v) eluent system.

[0114] White solid. Yield: 89% (2.46 g); m.p. 100.3-101.6° C.; TLC: R.sub.f=0.42 (DCM:MeOH (9:0.3; v/v)); C.sub.23H.sub.22F3N.sub.3O.sub.4 (461.44), Monoisotopic mass: 461.16. UPLC (100% purity): t.sub.R=7.15 min, (M+H).sup.+462.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.63-2.79 (m, 5H), 3.00-3.16 (m, 2H), 3.22-3.39 (m, 3H), 3.93-4.05 (m, 1H), 3.63-3.75 (m, 1H), 6.12 (s, 1H), 6.62 (s, 1H), 6.66-6.78 (m, 2H), 7.16-7.28 (m, 1H), 7.32-7.48 (m, 5H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.0, 42.2, 45.5, 48.3, 48.5, 56.8, 108.8, 112.1, 114.2, 120.4 (q, J=256.8 Hz), 128.7, 128.9, 129.8, 130.2, 132.8, 150.2, 151.9, 165.1, 176.3.

Example 11. 1-(2-Oxo-1-phenyl-2-(4-(4-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0115] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-[4-(trifluoromethoxy)phenyl]piperazine (1.48 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.3; v/v) eluent system.

[0116] White solid. Yield: 83% (2.29 g); m.p. 102.3-103.5° C.; TLC: R.sub.f=0.43 (DCM:MeOH (9:0.3; v/v)); C.sub.23H.sub.22F3N.sub.3O.sub.4 (461.44), Monoisotopic mass: 461.16. UPLC (100% purity): t.sub.R=7.17 min, (M+H).sup.+462.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.61-2.73 (m, 5H), 2.98-3.13 (m, 2H), 3.20-3.37 (m, 3H), 3.91-4.08 (m, 1H), 3.63-3.75 (m, 1H), 6.13 (s, 1H), 6.60 (s, 1H), 6.63-6.79 (m, 2H), 7.14-7.28 (m, 1H), 7.29-7.51 (m, 5H).

Example 12. 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl(sulfanyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0117] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-[3-(trifluoromethylthio)phenyl]piperazine (1.57 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0118] White solid. Yield: 64% (1.83 g); m.p. 97.8-99.2° C.; TLC: R.sub.f=0.48 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.22F.sub.3N.sub.3O.sub.3S (477.50), Monoisotopic mass: 478.13. UPLC (99% purity): t.sub.R=7.55 min, (M+H)+478.1. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.64-2.78 (m, 5H), 3.01-3.07 (m, 1H), 3.09-3.15 (m, 1H), 3.24-3.32 (m, 2H), 3.34 (dd, J=7.7, 3.2 Hz, 1H), 3.62-3.75 (m, 1H), 3.99 (ddd, J=13.2, 5.7, 3.4 Hz, 1H), 6.11 (s, 1H), 6.92 (dd, J=8.0, 2.3 Hz, 1H), 7.06 (s, 1H), 7.12 (d, J=7.4 Hz, 1H), 7.24-7.29 (m, 1H), 7.33-7.38 (m, 3H), 7.43 (d, J=6.8 Hz, 2H). .sup.13C NMR (126 MHz, CDCl.sub.3) δ 28.1, 45.6, 48.4, 48.7, 56.9, 118.5, 123.7, 125.3, 127.8, 128.5, 129.4 (d, J=141.2 Hz), 129.6 (d, J=137.0 Hz), 130.9, 132.9, 151.4, 165.2, 176.4.

Example 13. 1-(2-(4-(f1,1′-Biphenyl]-3-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0119] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-(biphen-3-yl)piperazine (1.43 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0120] White solid. Yield: 82% (2.23 g); m.p. 114.1-115.4° C.; TLC: R.sub.f=0.4 (DCM:MeOH (9:0.5; v/v)); C.sub.28H.sub.27N.sub.3O.sub.3 (453.54) Monoisotopic mass: 453.20. UPLC (100% purity): t.sub.R=7.43 min, (M+H).sup.+454.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.56-2.81 (m, 5H), 3.00-3.21 (m, 2H), 3.23-3.56 (m, 3H), 3.65-3.79 (m, 1H), 3.94-4.11 (m, 1H), 6.14 (s, 1H), 6.83 (dd, 1H, J=8.1, 2.0 Hz), 7.00-7.17 (m, 2H), 7.27-7.62 (m, 11H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.1, 42.5, 45.8, 49.0, 49.2, 56.8, 115.4, 115.6, 119.7, 127.2, 127.4, 128.7, 128.8, 129.6, 129.9, 132.9, 141.4, 142.5, 151.1, 165.0, 176.4.

Example 14. 1-(1-(4-Fluorophenyl)-2-oxo-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione

[0121] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-(4-fluorophenyl)acetic acid (1.51 g, 6 mmol, 1 eq) and 1-[3-(trifluoromethoxy)phenyl]piperazine (1.38 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0122] White solid. Yield: 73% (2.03 g); m.p. 88.8-90.7° C.; TLC: R.sub.f=0.63 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.21F.sub.4N.sub.3O.sub.3 (463.43), Monoisotopic mass: 463.15. UPLC (100% purity): t.sub.R=7.05 min, (M+H).sup.+464.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.61-2.89 (m, 5H), 3.02-3.46 (m, 5H), 3.67-3.80 (m, 1H), 3.88-4.04 (m, 1H), 6.09 (s, 1H), 6.94-7.27 (m, 6H), 7.29-7.40 (m, 2H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.0, 42.3, 45.6, 48.5, 48.6, 56.0, 112.7 (q, J=3.4 Hz), 115.9, 116.2, 116.7 (q, J=3.4 Hz), 117.0, 119.3, 124.1 (q, J=272.9 Hz), 125.5 (d, J=3.4 Hz), 129.7, 130.2, 130.3, 131.5 (q, J=31.1 Hz), 135.1 (d, J=6.9 Hz), 150.7, 160.9, 164.2, 164.5, 176.23.

Example 15. 1-(2-(4-(Naphth-2-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0123] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-(naphth-2-yl)piperazine (1.27 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0124] White solid. Yield: 79% (2.02 g); m.p. 197.1-198.5° C.; TLC: R.sub.f=0.71 ((DCM:MeOH (9:0.5; v/v)); C.sub.26H.sub.25N.sub.3O.sub.3 (427.50), Monoisotopic mass: 427.19. UPLC (100% purity): t.sub.R=6.97 min (M+H).sup.+428.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.58-2.87 (m, 5H), 3.03-3.25 (m, 2H), 3.29-3.60 (m, 3H), 3.69-3.89 (m, 1H), 3.96-4.17 (m, 1H), 6.12-6.18 (m, 1H), 7.00-7.24 (m, 2H), 7.28-7.54 (m, 7H), 7.61-7.80 (m, 3H).

Example 16. 1-(2-(4-(Benzo[b]thiophen-5-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0125] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-(benzo[b]thiophen-5-yl)piperazine (1.30 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0126] White solid. Yield: 79% (2.05 g); m.p. 164.1-165.3° C.; TLC: R.sub.f=0.66 ((DCM:MeOH (9:0.5; v/v)); C.sub.24H.sub.23N.sub.3O.sub.3S (433.53), Monoisotopic mass: 433.15 UPLC (100% purity): t.sub.R=6.62 min, (M+H).sup.+434.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.56-2.85 (m, 5H), 2.96-3.17 (m, 2H), 3.20-3.54 (m, 3H), 3.66-3.87 (m, 1H), 3.96-4.12 (m, 1H), 6.14 (s, 1H), 6.99 (dd, J=8.7, 1.9 Hz, 1H), 7.15-7.25 (m, 1H), 7.30-7.55 (m, 7H), 7.72 (d, J=8.8 Hz, 1H).

Example 17. 1-(2-(4-(1,2-Benzoxazol-5-il)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0127] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 5-(piperazin-1-yl)benzo[d]isoxazole (1.22 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0128] White solid. Yield: 57% (1.43 g); m.p. 186.4-187.8° C.; TLC: R.sub.f=0.58 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.22N.sub.4O.sub.4 (418.45), Monoisotopic mass: 418.16. UPLC (98% purity): t.sub.R=7.25 min, (M+H).sup.+419.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.57-2.86 (m, 5H), 2.95-3.19 (m, 3H), 3.22-3.53 (m, 2H), 3.62-3.84 (m, 2H), 3.94-4.11 (m, 1H), 6.14 (s, 1H), 7.05-7.32 (m, 1H), 7.29-7.54 (m, 6H), 7.98 (d, J=8.8 Hz, 1H).

Example 18. 1-(2-(4-(3-Chlorophenyl)piperidin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione

[0129] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 4-(3-chlorophenyl)piperidine (1.17 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0130] White solid. Yield: 74% (1.83 g); m.p. 111.8-113.4° C.; TLC: R.sub.f=0.43 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.23ClN.sub.2O.sub.3 (410.90), Monoisotopic mass: 410.14. UPLC (100% purity): t.sub.R=7.05 min, (M+H).sup.+411.1, .sup.1H NMR (300 MHz, CDCl.sub.3) δ 1.52-2.05 (m, 4H), 2.33-2.84 (m, 8H), 2.96-3.34 (m, 1H), 6.15 (s, 1H), 7.05-7.28 (m, 6H), 7.32-7.66 (m, 3H).

Example 19. 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperidin-1-yl)ethyl)pyrrolidine-2,5-dione

[0131] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 1-[3-(trifluoromethyl)phenyl]piperidine (1.37 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0132] White solid. Yield: 85% (2.26 g); m.p. 100.1-101.5° C.; TLC: R.sub.f=0.45 (DCM:MeOH (9:0.5; v/v)); C.sub.24H.sub.23F.sub.3N.sub.2O.sub.3 (444.45), Monoisotopic mass: 444.17. UPLC (100% purity): t.sub.R=7.26 min, (M+H).sup.+445.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 1.49-2.00 (m, 3H), 2.54-2.83 (m, 8H), 2.94-3.77 (m, 2H), 6.14 (s, 1H), 7.09-7.60 (m, 9H).

Example 20. 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperidin-1-yl)ethyl)pyrrolidine-2,5-dione

[0133] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 eq) and 4-[3-(trifluoromethoxy)phenyl]piperidine (1.45 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0134] White solid. Yield: 79% (2.18 g); m.p. 112.1-113.2° C.; TLC: R.sub.f=0.47 (DCM:MeOH (9:0.5; v/v)); C.sub.24H.sub.23F.sub.3N.sub.2O.sub.4 (460.45), Monoisotopic mass: 460.16. UPLC (100% purity): t.sub.R=7.12 min, (M+H).sup.+461.1, .sup.1H NMR (300 MHz, CDCl.sub.3) δ 1.38-2.15 (m, 3H), 2.49-2.92 (m, 8H), 2.99-3.85 (m, 2H), 6.15 (s, 1H), 7.11-7.64 (m, 9H).

Example 21. 1-(1-Oxo-3-phenyl-1-(4-phenylpiperazin-1-yl)prop-2-yl)pyrrolidine-2,5-dione

[0135] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 eq) and 1-phenylpiperazine (0, 97 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0136] White solid. Yield: 87% (1.13 g); m.p. 121.7-123.2° C.; TLC: R.sub.f=0.62 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.25N.sub.3O.sub.3 (391.47), Monoisotopic mass: 392.19. UPLC (100% purity): t.sub.R=6.22 min_(M+H).sup.+392.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.49-2.61 (m, 4H), 3.08 (d, J=16.4 Hz, 4H), 3.31-3.89 (m, 6H), 5.19 (dd, J=10.3, 6.1 Hz, 1H), 6.83-6.96 (m, 3H), 7.13-7.33 (m, 7H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 27.8, 34.2, 42.5, 45.4, 49.3, 49.6, 52.9, 116.6, 120.7, 127.1, 128.6, 129.1, 129.3, 136.7, 150.7, 166.4, 176.5.

Example 22. 1-(1-(4-(3-Chlorophenyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl)pyrrolidine-2,5-dione

[0137] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 eq) and 1-(3-chlorophenyl)piperazine (1.40 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0138] White solid. Yield: 87% (1.23 g); m.p. 114.3-116.2° C.; TLC: R.sub.f=0.80 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.24ClN.sub.3O.sub.3 (425.91), Monoisotopic mass: 426.15. UPLC (100% purity): t.sub.R=6.97 min, (M+H).sup.+426.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.49-2.64 (m, 4H), 3.07 (d, J=15.3 Hz, 4H), 3.30-3.86 (m, 6H), 5.17 (dd, J=10.1, 6.2 Hz, 1H), 6.73 (ddd, J=8.3, 2.2, 0.9 Hz, 1H), 6.79-6.88 (m, 2H), 7.06-7.35 (m, 6H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 27.8, 34.2, 42.2, 45.2, 48.7, 49.0, 52, 9, 114.4, 116.3, 120.2, 127.1, 128.6, 129.1, 130.2, 135.0, 136.6, 151.7, 166.4, 176.5.

Example 23. 1-(1-Oxo-3-phenyl-1-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)propan-2-yl)pyrrolidine-2,5-dione

[0139] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 eq) and 1-[3-(trifluoromethyl)phenyl]piperazine (1.38 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0140] White solid. Yield: 84% (1.59 g); m.p. 126.1-127.2° C.; TLC: R.sub.f=0.72 (DCM:MeOH (9:0.5; v/v)); C.sub.24H.sub.24F.sub.3N.sub.3O.sub.3 (459.47), Monoisotopic mass: 460.18. UPLC (100% purity): t.sub.R=7.22 min, (M+H).sup.+460.1. 1H NMR (300 MHz, CDCl3) δ 2.50-2.64 (m, 4H) 3.12 (d, J=14.8 Hz, 4H), 3.32-3.89 (m, 6H) 5.18 (dd, J=9.9, 6.2 Hz, 1H), 6.94-7.42 (m, 9H).

Example 24. 1-(1-(4-([1,1′-Biphenyl]-3-yl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl)pyrrolidine-2,5-dione

[0141] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 eq) and 1-(biphenyl-3)piperazine (1.43 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0142] White solid. Yield: 88% (1.46 g); m.p. 119.1-120.0° C.; TLC: R.sub.f=0.77 (DCM:MeOH (9:0.5; v/v)); C.sub.29H.sub.29N.sub.3O.sub.3 (467.57), Monoisotopic mass: 467.22. UPLC (100% purity): t.sub.R=7.63 min, (M+H).sup.+468.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.50-2.63 (m, 4H) 3.16 (d, J=18.9 Hz, 4H), 3.32-3.91 (m, 6H), 5.20 (dd, J=10.2, 6.0 Hz, 1H) 6.88 (dd, J=8.1, 1.8 Hz, 1H), 7.06-7.38 (m, 9H), 7.39-7.48 (m, 2H), 7.51-7.62 (m, 2H). .sup.13C NMR (75 MHz, CDCl.sub.3) δ 27.8, 34.2, 42.5, 45.4, 49.4, 49.7, 53.0, 115.5, 115.7, 119.7, 127.1, 127.2, 127.4, 128.6, 128.7, 129.1, 129.6, 136.7, 141.4, 142.5, 151.1, 166.4, 176.5.

Example 25. 1-(1-Oxo-3-phenyl-1-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)propan-2-yl)pyrrolidine-2,5-dione

[0143] The compound was prepared according to procedure described in Example 2. 2-(2,5-Dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 eq) and 1-[3-(trifluoromethoxy)phenyl]piperazine (1.48 g, 6 mmol, 1 eq) were used as starting materials. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0144] White solid. Yield: 83% (1.32 g); m.p. 104.4-105.5° C.; TLC: R.sub.f=0.71 (DCM:MeOH (9:0.5; v/v)); C.sub.24H.sub.24F.sub.3N.sub.3O.sub.4 (475.47), Monoisotopic mass: 475.17. UPLC (100% purity): t.sub.R=7.40 min, (M+H).sup.+476.1. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.50-2.63 (m, 4H, 3.10 (d, J=16.4 Hz, 4H), 3.31-3.86 (m, 6H), 5.18 (dd, J=9.9, 6.2 Hz, 1H), 6.62-6.85 (m, 3H), 7.11-7.36 (m, 6H).

Example 26. Determination of In Vivo Anticonvulsant Activity in Mice

[0145] The male Swiss albino mice (CD-1) weighing 18-26 g were used. All procedures were carried out in accordance with applicable Polish and international guidelines on the ethics of animal testing, after obtaining appropriate institutional approval. The substances were administered intraperitoneally (i.p.), in 1% aqueous solution of Tween, as single injections with a volume of 10 ml/kg, 30 minutes before the given test. Screening was performed on groups consisted of 4 mice. The average effective dose (ED.sub.50) in given test and toxic dose in the rotarod test (TD.sub.50) was estimated based on the results obtained in 3-4 groups of animals consisting of 6 mice. All tests were carried out based on the procedures described in the specialist literature.

Example 27. Maximal Electroshock Seizure Test (MES)

[0146] In the MES test, the seizures were induced by an electrical stimulus lasting of 0.2 s duration, 500 V voltage and 25 mA intensity. The electrical stimuli was generated using an electric shock generator (Rodent shocker, Type 221, Hugo Sachs Elektronik, Germany) and delivered to the animal using electrodes placed on the auricles. The study was conducted 30 minutes after intraperitoneal administration of the compounds at various doses. During the experiment, the number of animals that experienced a seizure episode in the form of tonic extension of the hind limbs was counted (Kaminski, K.; Rapacz, A.; fuszczki, J. J.; Latacz, G.; Obniska, J.; Kieć-Kononowicz, K.; Filipek, B. Bioorg. Med. Chem. 2015, 23, 2548-2561; Castel-Branco, M. M.; Alves, G. L.; Figueiredo, I. V.; Falcão, A. C.; Caramona, M. M. Methods Find. Exp. Clin. Pharmacol. 2009, 31, 101-106).

Example 28. Psychomotor Seizure Test (6 Hz Test)

[0147] In the 6 Hz test, seizures were induced by an electric stimulus of 32 mA and/or 44 mA and a frequency of 6 pulses per second. An electrical pulse was generated using an electric shock generator (ECT Unit 57800; Ugo Basile, Gemonio, Italy) and delivered to the animal using ocular electrodes. Before starting the test, the eye surface was gently moistened with a solution of local anesthetic (1% lidocaine solution). The study was conducted 30 minutes after intraperitoneal administration of the compounds at various doses. An electrical pulse was delivered continuously for a period of 3 seconds, followed by observation of the animal for a period of 10 seconds. During this time, immobility or stun associated with rearing, forelimb clonus, twitching of the vibrissae and Straub's tail were observed. These symptoms persist throughout the observation period, indicating the occurrence of psychomotor seizures in mice. Mice resuming normal behavior within 10 s after stimulation were considered as protected (Barton, M. E.; Klein, B. D.; Wolf, H. H.; White, H. S. Epilepsy Res. 2001, 47, 217-227; Wojda, E.; Wlaf, A.; Patsalos, P. N.; fuszczki, J. J. Epilepsy Res. 2009, 86, 163-174).

Example 29. Subcutaneous Pentylenetetrazole Seizure Test (scPTZ)

[0148] In the scPTZ test, seizures were induced by subcutaneous administration of pentylenetetrazole (PTZ) at a dose of 100 mg/kg. This caused clonic seizures with accompanying loss of the righting reflex. Test compounds were administered 30 minutes before the experiment. After PTZ administration, the animals were placed individually in transparent containers and observed for a period of 30 minutes for the occurrence of clonic seizures. During this time, the latency of the first onset of clonic seizures, defined as clonus of the whole body lasting at least 3 seconds with loss of the righting reflex and the number of seizure episodes during the test were noted and compared with control group. The absence of clonic convulsions within the observed time period was interpreted as the compound's ability to protect against PTZ-induced seizures (Ferreri, G.; Chimirri, A.; Russo, E.; Gitto, R.; Gareri, P.; De Sarro, A.; De Sarro, G. Pharmacol. Biochem. Behav. 2004, 77, 85-94; czkowski, K.; Saat, K.; Misiura, K.; Podkowa, A.; Malikowska, N. J. Enzyme Inhib. Med. Chem. 2016, 31, 1576-82).

Example 30. Influence on Mouse Motor Coordination in the Rotarod Test

[0149] The influence of tested compounds on motor coordination was assessed in the rotarod test (the apparatus used—May Commat, RR 0711 Rota Rod, Turkey). Mice were trained the day before the actual experiment. They were placed individually on a 2 cm diameter rod rotating at 10 revolutions per minute (rpm). During each training session, the animals remained on the rod for 3 minutes. The experiment was carried out 30 minutes after administration of the compounds. Motor coordination was tested at the speed of the rotating bar: 10 rpm during 60 seconds. Motor impairments were defined as the inability to remain on the rotating rod for 1 min. The mean time spent on the rod was counted in each experimental group (Dunham, N. W.; Miya, T. A.; Edwards, L. D. J. Am. Pharm. Assoc. 1957, 46, 64-66, Łczkowski, K.; Sałat, K.; Misiura, K.; Podkowa, A.; Malikowska, N. J. Enzyme Inhib. Med. Chem. 2016, 31, 1576-82).

Example 31. Statistical Analysis

[0150] The ED.sub.50 (effective dose) and TD.sub.50 (toxic dose) values along with the corresponding 95% confidence intervals were calculated based on the Litchfield and Wilcoxon method (Litchfield, J. T., Wilcoxon, F., 1949. A simplified method of evaluating dose-effect experiments. J. Pharmacol. Exp. Ther. 96, 99-113). To perform a statistical evaluation of the results, one-way ANOVA variance analysis and Dunnett's post hoc test (multiple comparison test) were used. The value at the significance level p<0.05 was considered statistically significant.

Example 32. Results of Anticonvulsant Activity Studies

[0151] The compounds of the invention showed broad anticonvulsant activity by acting effectively in the MES test, 6 Hz (32 mA and/or 44 mA) and scPTZ at a dose of 100 mg/kg. At time point of 30 min. they protected from 50-100% of the animals tested. The most potent protection revealed compounds containing electron withdrawing substituents at position 3 of the aromatic ring connected to piperazine moiety, preferably Cl, CF.sub.3, OCF.sub.3, SCF.sub.3, CHF.sub.2 or phenyl substituent, for which k is preferably 0. Table 1 shows pharmacological screening data for selected substances.

TABLE-US-00001 TABLE 1 Data from screening studies at a dose of 100 mg/kg for selected compounds according to the general formula (II). Test* Compound MES 6 Hz (32 mA) 6 Hz (44 mA) scPTZ 2 4/4 3/4 2/4 3/4 3 4/4 4/4 3/4 3/4 4 3/4 4/4 — 3/4 5 3/4 4/4 2/4 3/4 6 4/4 4/4 4/4 4/4 7 3/4 3/4 — 3/4 8 4/4 3/4 — 4/4 19 4/4 4/4 2/4 4/4 10 4/4 4/4 3/4 2/4 11 3/4 3/4 — 2/4 12 4/4 4/4 3/4 4/4 13 4/4 4/4 4/4 2/4 14 4/4 3/4 — 2/4 15 2/4 3/4 — 3/4 16 3/4 3/4 — 3/4 17 3/4 3/4 — 3/4 18 3/4 3/4 — 3/4 19 3/4 4/4 2/4 3/4 20 4/4 3/4 — 2/4 *Tests carried out in mice after intraperitoneal administration at a time point of 0.5 h, data indicate the number of mice protected in a given seizure model/number of mice tested; MES—the maximal electroshock test, the 6 Hz (32 mA) and 6 Hz (44 mA) test—the psychomotor seizures induced by low-frequency current (6 Hz) and intensity of 32 mA or 44 mA, respectively; scPTZ—the subcutaneous seizure test; “—”—substance not tested.

[0152] The above tests were carried out for racemic mixtures of compounds according to the invention.

[0153] Table 2 presents quantitative pharmacological data for selected compounds according to general formula (II), in particular for the selected active compound-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione (6), which protected 100% of the mice in the MES test, 6 Hz (32 mA and 44 mA) test and scPTZ test during the screening studies (time point of 0.5 h).

TABLE-US-00002 TABLE 2 The ED.sub.50 and TD.sub.50 values for selected compounds according to the general formula (II) and the model AED—valproic acid (VPA) after intraperitoneal administration in mice. ED.sub.50 [mg/kg] 6 Hz 6 Hz TD.sub.50 PI Compound MES (32 mA) (44 mA) scPTZ (mg/kg) (TD.sub.50/ED.sub.50) 2 91.1 83.5 — 100.0 >300 >3.3 (MES) >3.6 (6 Hz, 32 mA) 3 36.9 39.5 — 52.6 143.8 >3.0 (scPTZ) 3.9 (MES) 3.6 (6 Hz) 4 68.5 17.7 — >100 >300 2.7 (scPTZ) >4.4 (MES) 5 37.2 35.5 — 57.6 171.0 >17.0 (6 Hz, 32 mA) 4.6 (MES) 4.8 (6 Hz, 32 mA) 6 23.7 22.4 73.2 59.4 195.7 3.0 (scPTZ) 8.2 (MES) 8.7 (6 Hz, 32 mA) 9 97.7 63.0 195.7 94.3 274.2 2.7 (6 Hz, 44 mA) 3.3 (scPTZ) 2.8 (MES) 4.4 (6 Hz, 32 mA) 1.4 (6 Hz, 44 mA) 2.9 (scPTZ) 1.8 (MES) 10 41.7 38.3 — <60 74.0 1.9 (6 Hz, 32 mA) >1.2 (scPTZ) 4.1 (MES) 9.5 (6 Hz, 32 mA) 12 36.2 15.8 57.9 >100 150.0 2.6 (6 Hz, 44 mA) <1.5 (scPTZ) 1.7 (MES) 13 43.9 26.2 — <100 73.6 2.8 (6 Hz, 32 mA) >0.7 (scPTZ) 14 56.4 48.3 — <100 >300 5.3 (MES) 6.2 (6 Hz, 32 mA) 3.1 (MES) 19 81.8 41.0 — <100 254.3 6.2 (6 Hz, 32 mA) >2.5 (scPTZ) 1.7 (MES) 3.3 (6 Hz, 32 mA) VPA 252.7 130.6 183.1 239.4 430.7 2.3 (6 Hz, 44 mA) 1.8 (scPTZ) The substances were tested 0.5 h after intraperitoneal administration; MES—the maximal electroshock test, the 6 Hz (32 mA) and 6 Hz (44 mA) test—the psychomotor seizures induced by low-frequency current (6 Hz) and intensity of 32 mA or 44 mA, respectively; scPTZ—the subcutaneous seizure test; TD.sub.50 values were obtained in the rotarod test; PI—protective index (TD.sub.50/ED.sub.50); “—”—substance not tested.

[0154] The obtained results confirmed that the compounds of the invention, especially compound 6, have a potent protective effect and distinctly more favorable protective indexes in comparison to the model AED—valproic acid. Notably, valproic acid is known to possess wide spectrum of therapeutic indications.

Example 33. Determination of Antinociceptive Activity in the In Vivo Studies in Mice

[0155] The tests were carried out using male white Swiss mice (CD-1) weighing 18-25 g. All procedures were carried out in accordance with Polish and international guidelines on ethics of animal testing, after obtaining appropriate institutional approval. The study group consisted of 8-10 animals. The tested and reference substances were administered intraperitoneally 30 minutes before given test as suspension in 1% aqueous solution of Tween. All tests/models were carried out based on the procedures described in the specialist literature: the formalin test (Beirith, A.; Santos, A. R.; Calixto, J. B.; Rodrigues, A. L.; Creczynski-Pasa, T. B. Eur. J. Pharmacol. 1998, 345, 233-245), the model of capsaicin-induced pain (Mogilski, S.; Kubacka, M.; Redzicka, A.; Kazek, G.; Dudek, M.; Malinka, W.; Filipek, B. Pharmacol. Biochem. Behav. 2015, 133, 99-110), the model of oxaliplatin-induced neuropathic pain—von Frey test (Sałat, K.; Cios, A.; Wyska, E.; Sałat, R.; Mogilski, S.; Filipek, B.; Wiȩckowski, K.; Malawska, B. Pharmacol. Biochem. Behav. 2014, 122, 173-181).

Example 34. Determination of Analgesic Activity in the Formalin Test

[0156] The pain was induced by the subplantar injection of 20 μL of 2.5% formalin solution into the mice right hind paw. The animals were placed in separate, transparent observation chambers for a period of 30 minutes. The measured value was the total licking and biting time of the paw to which the formalin solution was injected. The nociceptive reaction time was calculated for the first 5 minutes after formalin injection (first phase of the test—acute pain) and in the 15-20, 20-25 and 25-30 minutes time intervals after its administration (second phase of the test—inflammatory pain). The observed inhibition of nociceptive reaction—reduction of paw licking and biting time, was interpreted as the analgesic effect of the compound tested. Based on the results obtained, the ED.sub.50 dose was calculated (dose that reduces the nociceptive reaction time by 50%). The reference compound in this test was valproic acid, which was administered intraperitoneally at doses of 100 mg/kg, 150 mg/kg and 200 mg/kg. Compound 6 was administered at doses of 10 mg/kg, 20 mg/kg and 30 mg/kg.

[0157] Compound 6 showed distinct analgesic activity in both phases of the test. The mean nociceptive response time in the control group was 90.0±4.97 seconds and 212.70±10.16 seconds in the first and second phase of the test, respectively. Compound 6, at all doses tested, reduced the nociceptive reaction time in the first phase of the formalin test, corresponding to acute pain, with a statistically significant effect observed at the two highest doses. The ED.sub.50 value for compound 6 in the first phase of the test was 28.50 mg/kg. In the second phase of the test corresponding to tonic inflammatory pain, compound 6, at all doses used, statistically significantly shortened the time of nociceptive reaction. The ED.sub.50 value in the second phase of the test for this compound was 12.40 mg/kg (FIG. 3).

[0158] Valproic acid (VPA) showed no analgesic activity in the first phase of the test at any of the doses tested. In the second phase of the VPA test, the nociceptive response time was reduced at all doses used, and the ED.sub.50 value in this phase of the test was 132.90 mg/kg (FIG. 3).

Example 35. Determination of Analgesic Activity in the Capsaicin Pain Model

[0159] This test assesses the time of licking and/or biting the hind paw into which mice were injected subplantarly with capsaicin in an amount of 1.6 μg, dissolved in 20 μl of a mixture containing 0.9% saline and ethanol (5% of the final volume). Observation was carried out for minutes after capsaicin administration. Test compounds were administered intraperitoneally 30 minutes before administration of capsaicin. Inhibition of nociceptive reaction—shortening the time of licking and biting the paw was the measure of antinociceptive activity of the compound tested.

[0160] Valproic acid (VPA) was the reference compound in this test. VPA was administered intraperitoneally at doses of 100 mg/kg, 150 mg/kg and 200 mg/kg. Compound 6 was administered at 20 mg/kg, 30 mg/kg and 40 mg/kg. Test compounds were administered as a suspension in 1.0% Tween 80 solution. The control group consisted of mice treated with vehicle alone (1% Tween 80 solution). The nociceptive reaction time in this group was 43.29±3.21 seconds.

[0161] Compound 6 in statistically significant manner reduced the nociceptive response time at 20 mg/kg and 30 mg/kg, and the ED.sub.50 was 17.9 mg/kg (FIG. 4) The reference compound (valproic acid) statistically significantly reduced the nociceptive reaction time to 25.00±4.57 seconds (corresponding to an analgesic activity of 42.25%), only after the 200 mg/kg dose (FIG. 4).

Example 36. Determination of Analgesic Activity in the Model of Oxaliplatin-Induced Neuropathic Pain—Von Frey Test

[0162] Oxaliplatin (OXPT) was dissolved in a 5% glucose solution, followed by intraperitoneal administration to mice. A single dose of 10 mg/kg was used. Tactile and thermal (sensation of low temperature) allodynia associated with oxaliplatin-induced neuropathy are characterized by two phases. The early phase is acute and develops soon after administration of the OXPT, while the symptoms of the later (chronic) phase (associated with neurons damage), develops after a few days. Behavioral tests in mice with OXPT-induced neuropathy were carried out 7 days after its administration, i.e. in the late phase of neuropathy.

[0163] The influence of compounds tested on tactile allodynia was determined in the von Frey test. The animals were placed individually in cages with a reticulated bottom, 60 minutes before the beginning of the experiment, in order to adapt to the new environment. An electronic Von Frey apparatus (Electronic Von Frey, Bioseb, France) was used to assess the pain threshold for mechanical stimuli. The von Frey's fiber was applied to the underside of the right mouse paw with increasing pressure. The crossing of pain threshold resulted in the paw withdrawal and subsequent recording of the mechanical pressure that evoked the nocifensive response. The measurement was performed 3 times for each mouse, with at least 30 seconds between the measurements, then the results obtained were averaged. The entire study was performed 3 times: before OXPT administration to determine the baseline pain threshold; 7 days after administration of OXPT, and before administration of test compounds, to assess developing neuropathy by setting a new pain threshold; 30 minutes after administration of the compounds to determine their influence developed neuropathy.

[0164] The effect of the test compound on thermal allodynia was evaluated in a Cold Plate test using specialized equipment—Cold/Hot Plate; Bioseb, France. The animals were placed individually on a metal plate cooled to 2° C. using the thermostated device. The observed nociceptive reactions of animals included licking and/or characteristic hind paw lift or bounce. The observation time was set at 60 s to eliminate the potential risk of tissue damage and minimize animal discomfort. Similar to the Von Frey test, the measurement was performed 3 times.

[0165] Compound 6 and valproic acid as a reference AED were administered intraperitoneally as a suspension in a 1% solution of Tween 80. Compound 6 was administered at doses of 10, 20 and 30 mg/kg. The reference compound (valproic acid) was given at doses of 50, 100 and 150 mg/kg.

[0166] Injection of OXPT in mice caused development of neuropathy resulting in a prominent and statistically significant reduction in the pain threshold as measured by the von Frey method. The pain sensitivity threshold decreased from 3.18±0.06-3.36±0.10 g in healthy mice to a level in the range 1.89±0.04-1.94±0.14 g in OXPT-administered mice. The obtained results indicate a statistically significant analgesic effect of the tested compound 6. The average pain sensitivity threshold in the control group was 3.36±0.10 g, whereas after the administration of OXPT it decreased to 1.89±0.04 g (56.25% of the initial value). Administration of compound 6 at a dose of 10 mg/kg increased the pain threshold to 2.87±0.12 g (85.41% of the initial value), which indicates the inhibitory effect on the development of mechanical allodynia already at low doses. The dose of 20 mg/kg of compound 6 caused an increase in pain sensitivity threshold to 3.83±0.13 g, which is 113.98% of the initial value. The 30 mg/kg dose resulted in an increase in pain threshold to 4.17±0.17 g, which is 124.10% of the initial value. The results obtained indicate that compound 6 is highly effective in suppressing of mechanical allodynia development which is the result of neurons damage caused by the chemotherapeutic agent—OXPT (FIG. 5A).

[0167] The average pain sensitivity threshold in the control group for the reference compound (valproic acid, VPA) was 2.62±0.06 g, and after the administration of OXPT it decreased to 1.78±0.04 g. Administration of VPA at a dose of 150 mg/kg caused an increase in the pain threshold up to 3.97±0.30 g, while doses of 100 mg/kg and 50 mg/kg body weight allowed to achieve an increase in the average pain threshold to 3.18±0.14 g and 2.75±0.06 g, respectively (FIG. 5B).

[0168] Compound 6 also significantly increased thermal allodynia sensitivity in the cold plate test (FIG. 5C).

Example 37. In Vitro Affinity and Functional Studies

[0169] The affinity and functional tests performed in vitro for the most active substance 6, representing compounds according to formula (II), showed that their mechanism of action is associated with the effect on neuronal conductivity through interaction with voltage-dependent sodium channels (site 2) and calcium channels (dihydropyridine, diltiazem and verapamil binding sites). A unique feature of the compound 6 representing compounds of formula (II) according to the invention is the inhibition of calcium currents by blocking the transient receptor potential vanilloid type 1 (TRPV1). This effect has not been disclosed for known AEDs. TRPV1 receptor antagonism may determine the antinociceptive effect of the compounds disclosed herein. The role of the TRPV1 receptor in conduction of pain stimuli has been well documented in specialist literature (Szallasi, A.; Cortright, D. N.; Blum, C. A.; Eid, S. R. Nat. Rev. Drug. Discov. 2007, 6, 357-372). The compounds according to the invention are characterized by a complex mechanism of action, which is not described for known anticonvulsants. It should be emphasized, however, that further in vitro studies may reveal further molecular targets responsible for the pharmacological action of substances being the subject of current patent claim. The results of binding studies (sodium channel, calcium channel) and functional tests (TRPV1 receptor) for compound 6 are shown in Table 3.

TABLE-US-00003 TABLE 3 In vitro affinity/functional tests results for compound 6, representing substances according to formula (II) of the invention* Inhibition of specific control binding Affinity studies Material (concentration tested [μM]).sub.g Na.sub.+ channel (miejsce 2).sub.a Rat cerebral cortex 82.5 (100) 33.4 (10) Ca.sub.2+ channel type L Rat cerebral cortex 82.3 (100) (dihydropyridine binding site).sub.b 30.8 (10) Ca.sub.2+ channel type L Rat cerebral cortex 69.6 (100) (diltiazem binding site).sub.c Ca.sub.2+ channel type L Rat cerebral cortex 58.2 (100) (verapamil binding site).sub.d Potassium channel (hERG).sub.e Human recombinant 25.8 (100) cells (HEK-293) % Inhibition of agonist control responses Functional studies (concentration tested [μM]).sub.a TRPV1 receptor (VR1) (h) Human recombinant 71.7 (100) (antagonist effects).sub.f cells (CHO) *The tests were carried out in CEREP laboratories (France) according to the procedures described in the literature: .sub.aBrown, G. B. J. Neurosci. 1986, 6, 2064-2070; .sub.bGould, R. J.; Murphy, K. M.; Snyder, S. H. Proc. Natl. Acad. Sci. USA. 1982, 79, 3656-3660; .sub.cSchoemaker, H.; Langer, SZ. Eur. J. Pharmacol. 1985, 111, 273-277; .sub.dReynolds, I. J.; Snowman, A. M.; Snyder, S. H. J. Pharmacol. Exp. Ther. 1986, 237, 731-738; .sub.eHuang, X. P.; Mangano, T.; Hufeisen, S.; Setola, V.; Roth, B. L. Assay Drug Dev. Technol. 2010, 8, 727-742; .sub.fPhelps, P. T.; Anthes, J. C.; Correll, C. C. Eur. J. Pharmacol. 2005, 513, 57-66; .sub.g% inhibition >50% is considered as significant effect exerted by the compound.

Example 38. In Vitro Electrophysiological Studies

[0170] The experiments were carried out in accordance with institutional and international guidelines regarding the ethics of animal research. Rats (3 weeks old) were anesthetized with ethyl chloride and decapitated. The brains were then removed and placed in ice-cold extracellular fluid. The methodology of slice preparation and pre-incubation has been described earlier (Szulczyk, B.; Nurowska, E. Biochem. Biophys. Res. Commun. 2017, 491, 291-295). The sections containing the prefrontal cortex were enzymatically and mechanically dispersed. Single prefrontal cortex pyramidal neurons were visualized using an inverted microscope (Nikon). Sodium currents were induced by rectangular depolarizing stimuli. The potential between depolarizing stimuli was maintained at −65 mV.

[0171] The intracellular fluid in the pipette contained (in mM): CsF (110), NaCl (7), EGTA (3), HEPES-CI (10), MgCl.sub.2 (2), Na.sub.2ATP (4) (pH 7.4 and osmolarity 290 mOsm).

[0172] The extracellular fluid washing the neurons contained (in mM): NaCl (30), choline chloride (90), TEA-Cl (30), CaCl.sub.2 (2), MgCl.sub.2 (2), glucose (15), HEPES (10), LaCl.sub.3 (0.001) and CdCl.sub.2 (0.4) (pH 7.4). Currents were recorded using an Axopatch 1D amplifier and analyzed using pClamp software (Axon Instruments and Molecular Devices, USA). Pipette resistance was between 4 and 5 MΩ. After gigaseal formation, pipette capacitance was compensated by means of an amplifier.

[0173] The patch membrane was ruptured by suction or by an electrical stimulus, and then membrane capacitance was compensated. Access resistance was between 5 and 7 MΩ. A series resistance compensation of 80% was used. The leakage current was subtracted from the recorded currents. Recordings were carried out at room temperature. Voltage-dependent potassium currents were blocked by TEA-Cl in the extracellular fluid. Voltage-dependent calcium currents were blocked by cadmium and lanthanum ions in the extracellular fluid. The neuron's membrane potential was maintained at −65 mV. Substance 6 was administered from the extracellular side (to the whole bath).

[0174] The obtained results confirmed the inhibitory effect of compound 6 on rapidly activating and rapidly inactivating voltage-dependent sodium channels in the prefrontal cortex pyramidal neurons (tests were performed at a concentration of 100 μM). Maximum currents were induced by rectangular depolarizing stimuli lasting 20 msec. The potential between depolarizing stimuli was maintained at −65 mV. Control recordings were carried out for 2 minutes, the test substance was administered for 3 minutes and the currents after washing out were recorded for 5 minutes. Recorded currents were normalized to the value of control currents. Substance 6 blocked the maximum amplitude of sodium currents up to 0.59±0.08 compared to the control (1.0, p<0.001). After washing out, the current amplitude partially returned to control values (0.79±0.07, n=5). Examples of sodium current recordings and averaged results are shown in FIG. 6.

Example 39. Evaluation of ADMETox Parameters in In Vitro Studies

[0175] The ADME-Tox parameters of compound 6 were estimated by in vitro methods using recombinant enzymes, human and mouse liver microsomes, and eukaryotic cell lines.

[0176] Metabolic stability. The metabolic stability of compound 6 was evaluated using human liver microsomes (HLMs). Internal clearance values CL.sub.int were calculated by monitoring changes in compound concentration in the presence of microsomes per unit of time, according to the procedure proposed by Obach R. S. (Obach, R. S. Drug Metab. Dispos. 1999, 27, 1350-1359). Based on the data obtained, an extremely low clearance value of compound 6 after incubation with HLMs was found, amounting to CL.sub.int=5.8 ml/min/kg, indicating its predicted high stability in the human body. UPLC analysis of the metabolism of compound 6 after incubation with HMLs revealed that it is metabolized to three metabolites M1-M3 (FIG. 7). Based on the UPLC/MS data, it was found that the metabolite M1 is formed by dehydrogenation of the piperazine ring, M2 is formed by hydroxylation of the phenyl substituent linked to piperazine, while M3 is probably formed as a result of hydroxylation of the side phenyl group with simultaneous reduction of the ketone to hydroxyl group in the imide fragment (FIG. 7).

[0177] Metabolic stability tests—methodology. Metabolic stability studies for compound 6 were performed using HLMs (Promega, Madison, Wis., USA). For this purpose, 10 μL of compound 6 at a concentration of 1000 μM was diluted with 132 μL with tris-HCl buffer (100 mM, pH 7.4), followed by the addition of 8 μL of appropriate microsomes. The reaction mixture was preincubated at 37° C. for 5 minutes, followed by the addition of 50 μL of NADPH Regeneration System, supplied by Promega (Madison, Wis., USA). After mixing, the whole mixture was incubated at 37° C. for 120 minutes. To complete the reaction, 200 μL of cold methanol was added to the tubes and centrifuged. The supernatant was subjected to UPLC/MS analysis, including fragmentation analysis. Four mixtures of 6 with HLMs were prepared to determine the internal clearance CL.sub.int. Each of these reactions was completed at a different time point, after 5, 15, 30 and 45 min, by the addition of cold methanol containing 50 μM internal standard. Then, according to literature guidelines (Obach, R. S. Drug Metab. Dispos. 1999, 27, 1350-1359), based on the plot of the relationship between the height of the peak from 6 and the height of the internal standard, the regression equation was determined and the reaction rate constant k was calculated. Then the constant k was substituted to equation (1).

[00001]$\begin{array}{cc}{t}_{1/2}=\frac{\ln 2}{-k}& \left(1\right)\end{array}$

[0178] The calculated t.sub.1/2 value was then substituted into equation (2):

[00002]$\begin{array}{cc}{\mathrm{CL}}_{\mathrm{int}\left(\mathrm{human}\right)}=\frac{0.693}{\mathrm{in}\mathrm{vitro}{t}_{1/2}}\times \frac{\begin{array}{c}\mathrm{mL}\mathrm{of}\\ \mathrm{mixture}\end{array}}{\begin{array}{c}\mathrm{mg}\mathrm{of}\\ \mathrm{microsomes}\end{array}}\times \frac{\begin{array}{c}45\mathrm{mg}\mathrm{of}\\ \mathrm{microsomes}\end{array}}{g\mathrm{of}\mathrm{liver}}\times \frac{\begin{array}{c}20g\mathrm{of}\\ \mathrm{liver}\end{array}}{\begin{array}{c}\mathrm{kg}\mathrm{of}\mathrm{body}\\ \mathrm{weight}\end{array}}& \left(2\right)\end{array}$

[0179] Impact on Pgp activity. P-glycoprotein (Pgp) is an integral plasma membrane protein that, as an ATP-dependent burst pump, actively removes xenobiotics and can cause drug interactions. Pgp plays an important role in the absorption of drugs in the gastrointestinal tract and also through the blood-brain barrier. A commercial bioluminescent Pgp-Glo™ Assay System test (Promega, Madison, Wis., USA) was used to study the effect of compound 6 on Pgp activity. The operation of the test is based on measuring changes in the level of ATP consumed by membranes containing the recombinant Pgp protein in the presence of test compounds. The results were presented as % of baseline activity and compared to reference compounds: selective Pgp inhibitor Na.sub.3VO.sub.4 and verapamil stimulator. Compound 6 showed a statistically significant (p<0.01) inhibitory effect on Pgp up to 38% of baseline activity at 100 μM, while no effect on Pgp activity at 50 μM was noted (FIG. 8)

[0180] Impact on Pgp activity—methodology. The tests were performed according to the protocol of the bioluminescent Pgp-Glo™ Assay System test provided by the Promega company (Madison, Wis., USA). Enzyme reactions were performed in Nunc™ MicroWell™ 96-well white plates from Thermo Scientific (Waltham, Mass., USA). Bioluminescence was measured with a PerkinElmer multispecific EnSpire plate reader (Waltham, Mass. USA). After using the Na.sub.3VO.sub.4 Pgp inhibitor (induces 100% inhibition), there was an increase in signal relative to the control sample, indicating inhibition of ATP consumption by Pgp, the so-called base activity. The calculated difference between the luminescence values of the inhibitor-treated sample and the control sample was taken as 100% of the Pgp base activity and treated as a negative control in the test. The reference compounds Na.sub.3VO.sub.4 and verapamil were used at 100 μM and 200 μM, respectively, according to the manufacturer's instructions. Compound 6 was tested at 50 and 100 μM concentrations obtained after dilution of concentrated stock solution (10 mM) in DMSO in reaction buffer. Incubation of the compounds with Pgp-containing membranes was carried out for 40 minutes at 37° C., followed by bioluminescence measurement to determine the degree of ATP consumption by Pgp. Statistical significance was calculated by one-way ANOVA variance analysis and the Bonferroni method using GraphPad Prism 5. The compounds were tested in triplicate.

[0181] Effect of compound 6 on cytochrome P-450 3A4 and 2D6 activity. The research was conducted using commercial luminescence tests CYP3A4 P450-Glo™ and CYP2D6 P450-Glo™ from Promega (Madison, Wis., USA) based on the methodology described in the literature (Socata, K.; Mogilski, S.; Pieróg, M.; Nieoczym, D.; Abram, M.; Szulczyk, B.; Lubelska, A.; Latacz, G.; Doboszewska, U.; Wlaź, P.; Kaminski, K. ACS Chem. Neurosci. 2018, doi: 10.1021/acschemneuro.8b00476; Latacz, G.; Lubelska, A.; Jastrzȩska-Wisek, M.; Partyka, A.; Sobato, A.; Olejarz, A.; Kucwaj-Brysz, K.; Satała, G.; Bojarski, A. J.; Wesołowska, A.; Kieć-Kononowicz, K.; Handzlik, J. Chem. Biol. Drug. Des. 2017, 90, 1295-1306). The CYP isoforms selected for study are responsible for the metabolism of approximately 40-50% of the drugs available on the market, and their stimulation or inhibition determines the majority of metabolic drug interactions. The results obtained indicate no effect of compound 6 on CYP3A4 activity (FIG. 9A) and a very weak stimulating effect on CYP2D6 (FIG. 9B) at a concentration of 10 μM. In summary, the results obtained indicate a low probability of potential metabolic interactions caused by 6.

[0182] In vitro hepatotoxicity assessment. The studies were conducted using the hepatoma HepG2 liver cancer cell line, which is used to assess the hepatotoxicity of the substance in vitro. A classic MTS colorimetric assay from Promega (Madison, Wis., USA) was used to investigate the effect of 6 on HepG2 cell viability and proliferation. The compound was tested at four concentrations in the range (0.1-100 μM). Doxorubicin at a concentration of 1 μM was used as a reference cytostatic. In addition, the reference mitochondrial toxin carbonyl cyanide m-chlorophenylhydrazone (CCCP) at a concentration of 10 μM was also used (FIG. 10). Hepatotoxicity studies after 72 h incubation of the HepG2 line with compound 6 showed a statistically significant (p<0.05) reduction in cell viability only for the highest concentration of 100 μM used in the study (FIG. 10). In addition, cell viability was reduced to just 84% of the control, indicating a trace toxic effect of this compound on HepG2 cell lines. Due to the particular exposure of liver cells to the potential toxic effects of xenobiotics, an additional test was carried out with the HepG2 line in the form of luminescent measurement of ATP levels in cells, after a short 3-hour exposure to compound 6, in concentrations in the range of 1-100 μM. For this purpose, a commercial CellTiter-Glo Luminescent Cell Viability Assay from Promega (Madison, Wis., USA) was used. The aim of the study was to check the effect of the compound on mitochondrial respiration of hepatoma cells. The reference point was the CCCP reference mitochondrial toxin at a concentration of 10 μM. There was no statistically significant effect of Compound 6 on ATP levels in HepG2 cells, even at the highest concentration of 100 μM used. This indicates a very low risk of hepatotoxic effect of compound 6 (FIG. 11).

[0183] In vitro hepatotoxicity assessment—methodology. HepG2 hepatoma cell line (ATCC HB-8065) was used for the studies. The HepG2 line was incubated in “Modified Eagle's Medium” (MEM) culture medium with the addition of 2 mM glutamine and 10% FBS from Gibco (Carlsbad, Calif., USA). Cells were incubated at 37° C. in an atmosphere containing 5% CO.sub.2. CellTiter 96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) supplied by Promega (Madison, Wis., USA) was used to test cell viability. Prior to testing, cells were placed on Thermo Scientific Nunc™ 96-well transparent culture plates (Waltham, Mass., USA) at a concentration of 1.5×10.sup.4 cells per well and incubated for 24 h. Then a 10 mM stock solution of compound 6 was diluted. in the appropriate culture medium and added to the cells at final concentrations in the range of 0.1-100 μM (DMSO concentration in all wells was 1%). Reference compounds CCCP and DX were applied at final concentrations of 10 μM and 1 μM, respectively. After 72 h incubation at 37° C. in an atmosphere containing 5% CO.sub.2, the medium with the compound was removed, followed by the addition of fresh medium with diluted MTS reagent. Plates were again incubated for 2-3 hours, followed by absorbance measurement at 490 nm with an EnSpire PerkinElmer (Waltham, Mass., USA) reader. Statistical significance was calculated by one-way ANOVA variance analysis and the Bonferroni method. The compounds were tested in four replications.

[0184] CellTiter-Glo Luminescent Cell Viability Assay from Promega (Madison, Wis., USA) was used to study ATP levels in HepG2 cells. Prior to testing, the cells were plated into white, 96-well transparent bottom culture plates from Corning (Tewksbury, Mass., USA), adapted for luminescence measurement at a concentration of 1.5×10.sup.4 cells per well. Then, the cells were incubated at 37° C. in an atmosphere containing 5% CO.sub.2. Compound 6 was applied to the plate at three final concentrations of 1, 10 and 100 μM, CCCP at 10 μM and DX at 1 μM, and the amount of 100 μL. The plate was incubated for 3 h at 37° C. and 5% CO.sub.2. Luminescence measurement was carried out with an EnSpire PerkinElmer (Waltham, Mass., USA) reader after adding CellTiter-Glo Luminescent Cell Viability Assay in an amount of 100 μl to the culture. Statistical significance was calculated by one-way ANOVA and Bonferroni analysis using GraphPad Prism 5. All substances were tested in four replications.

Example 40. Preparation of Selected Enantiomers of Compounds According to the Invention

[0185] Enantiomers of compounds according to formula (II) of the invention can be obtained applying four-stage procedure using commercially available tert-butoxycarbonyl (Boc) D- or L-amino acid derivatives (R or S absolute configuration, respectively) as starting materials. Enantiomers were obtained for selected compounds described by formula (II) for which k=0, A and B have the meaning as in the case of racemic mixtures of formula (II).

[0186] A general scheme for the synthesis of enantiomers of compounds according to formula (II) is shown in FIG. 12.

[0187] In the first stage, the condensation reaction of given piperazine derivative with the corresponding Boc-D- or Boc-L-amino acid derivative yields an intermediate product of formula (VII), which subsequently forms the amine derivative (VI) in the deprotection reaction. In the next step, compound (VI) is condensed with succinic anhydride to obtain the intermediate with the amide-acid structure (V), which next undergoes cyclization reaction to form compound R-(II) or S-(II). Asymmetric synthesis proceeds with retention of absolute configuration that was confirmed applying crystallographic analysis.

[0188] Examples of synthesis as well as physicochemical and spectral data for selected intermediates (VII, VI, and V according to FIG. 12) are described below.

Example 41. Tert-butyl-(R)-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)carbamate (VII)

[0189] Boc-D-phenylglycine (1.25 g, 5 mmol, 1 eq) was dissolved in 20 mL of DCM, followed by the addition of DCC (1.55 g, 7.5 mmol 1.5 eq). Next after 30 minutes 1-(3-(trifluoromethyl)phenyl)piperazine (1.15 g, 5 mmol, 1 eq) dissolved in 5 mL of DCM was added. The reaction was continued with stirring at room temperature for 4 hours. After this time, DCM was distilled off to dryness. Intermediate VII was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0190] Light oil. Yield: 78% (1.81 g); TLC: R.sub.f=0.62 (DCM:MeOH (9:0.5; v/v)); C.sub.24H.sub.28F.sub.3N.sub.3O.sub.3 (463.50), Monoisotopic mass: 463.21. UPLC (100% purity): t.sub.R=8.40 min. (M+H).sup.+464.2.

Example 42. (R)-2-Amino-2-phenyl-1-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethan-1-one (VI)

[0191] mL of TFA was added to the tert-butyl-(R)-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)carbamate solution (VII, 1.39 g, 3 mmol, 1 eq) in DCM (50 mL) and stirred for 2 hours. The reaction mixture was then neutralized with a 25% NH.sub.4OH solution, followed by extraction with DCM (3×50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and then evaporated to dryness. (R)-2-Amino-2-phenyl-1-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethan-1-one was obtained as a yellow oil.

[0192] Yellow oil. Yield: 95% (1.03 g); C.sub.19H.sub.20F.sub.3N.sub.3O (363.38), Monoisotopic mass: 363.16. UPLC (purity >99.9%): t.sub.R=4.96 min. (M+H).sup.+364.1.

Example 43. (R)-4-Oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl) ethyl)amino)butanoic Acid (V)

[0193] Succinic anhydride (0.28 g 2.8 mmol, 1 eq) was added to a solution of (R)-2-amino-2-phenyl-1-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethan-1-one (VI, 1.02 g 2.8 mmol, 1 eq) in AcOEt (50 mL) and the mixture was stirred for 30 minutes. After this time, the solvent was distilled off to dryness. The compound was obtained in solid form after washing with Et.sub.2O.

[0194] White solid. Yield: 87% (1.13 g); C.sub.23H.sub.24F.sub.3N.sub.3O.sub.4 (463.46), Monoisotopic mass: 463.17. UPLC (purity >99.9%): t.sub.R=6.40 min. (M+H).sup.+464.2.

Example 44. (R)-1-(2-(4-(3-chlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione ((R)-3)

[0195] ZnCl.sub.2 (0.27 g, 2.0 mmol, 1 eq) was added to the suspension of (R)-4-((2-(4-(3-chlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)amino)-4-oxobutanoic acid (V, 0.86 g, 2.0 mmol, 1 eq) in dry benzene (50 mL) and the whole mixture was heated to 80° C., then HMDS solution (0.48 g, 0.62 ml, 3.0 mmol, 1.5 eq) in dry benzene (5 ml) was added dropwise over 30 minutes. The reaction was continued with stirring in reflux for about 24 hours and then concentrated under reduced pressure. After distilling off the solvent, the oily residue was dissolved in DCM and extracted with 0.1 M HCl (3×50 mL), water (3×50 mL) and saturated NaCl solution (3×50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and then evaporated to dryness. The crude product was purified by column chromatography using DCM:MeOH (9:0.3; v/v) eluent system. The compound was obtained as a solid after washing with Et.sub.2O.

[0196] White solid. Yield: 82% (0.67 g); m.p. 167.3-168.1° C.; TLC: R.sub.f=0.41 (DCM:MeOH (9:0.3; v/v)); C.sub.22H.sub.22ClN.sub.3O.sub.3 (411.89), Monoisotopic mass: 411.13. UPLC (purity: >99.9%): t.sub.R=6.70 min, (M+H).sup.+412.4. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.64-2.75 (m, 5H), 2.96-3.12 (m, 2H), 3.21-3.37 (m, 3H), 3.60-3.72 (m, 1H), 3.92-4.03 (m, 1H), 6.10 (s, 1H), 6.68 (dd, J=8.0, 2.3 Hz, 1H), 6.77 (t, J=2.0 Hz, 1H), 6.81 (d, J=7.6 Hz, 1H), 7.13 (t, J=7.9 Hz, 1H), 7.32-7.37 (m, 3H), 7.42 (d, J=6.8 Hz, 2H). .sup.13C NMR (126 MHz, CDCl.sub.3) δ 28.1, 42.3, 45.6, 48.5, 48.8, 56.9, 114.5, 116.4, 120.3, 128.8, 129.0, 129.9, 130.2, 133.0, 135.1, 151.8, 165.1, 176.4. Enantiomeric purity >99% (t.sub.R=40.25 min).

Example 45. (R)-1-(2-(4-(3,5-dichlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione ((R)-4)

[0197] The compound was prepared according to procedure described in Example 44. (R)-4-((2-(4-(3,5-dichlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)amino)-4-oxobutanoic acid (0.93 g, 2 mmol, 1 eq) was used as the starting material for the cyclization reaction. The crude product was purified by column chromatography using DCM:MeOH (9:0.2; v/v) eluent system.

[0198] White solid. Yield: 79% (0.70 g); m.p. 174.3-175.5° C.; TLC: R.sub.f=0.43 (DCM:MeOH (9:0.2; v/v)); C.sub.22H.sub.21Cl.sub.2N.sub.3O.sub.3 (446.33), Monoisotopic mass: 445.10. UPLC (purity: >99.9%): t.sub.R=7.59 min, (M+H).sup.+446.1. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.64-2.74 (m, 5H,), 2.99-3.03 (m, 1H), 3.06-3.11 (m, 1H), 3.23-3.31 (m, 2H), 3.43-3.47 (m, 1H), 3.60-3.64 (m, 1H), 3.95-3.99 (m, 1H), 6.08 (s, 1H), 6.63 (d, J=1.7 Hz, 2H), 6.79 (t, J=1.4 Hz, 1H), 7.32-7.37 (m, 3H), 7.41 (d, J=6.7 Hz, 2H). .sup.13C NMR (126 MHz, CDCl.sub.3) δ 28.1, 42.1, 45.4, 47.9, 48.2, 56.8, 114.3, 119.8, 128.8, 129.1, 129.9, 132.9, 135.6, 152.1, 165.2, 176.4. Enantiomeric purity >99% (t.sub.R=43.23 min).

Example 46. (R)-1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl) pyrrolidine-2,5-dione ((R)-6)

[0199] The compound was prepared according to procedure described in Example 44. (R)-4-Oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)amino)-butanoic acid (0.93 g, 2.0 mmol, 1 eq) was used as the starting material for the cyclization reaction. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0200] White solid. Yield: 80% (0.71 g); m.p. 189.1-190.5° C.; TLC: R.sub.f=0.35 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.22F.sub.3N.sub.3O.sub.3 (445.44), Monoisotopic mass: 445.16. UPLC (purity: >99.9%): t.sub.R=6.93 min, (M+H).sup.+446.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.52-2.85 (m, 5H), 2.99-3.19 (m, 2H), 3.22-3.45 (m, 3H), 3, 62-3.76 (m, 1H), 3.93-4.07 (m, 1H), 6.12 (s, 1H), 6.90-7.15 (m, 3H), 7.11 (d, 1H, J=7.7 Hz), 7.28-7.55 (m, 6H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.0, 42.3, 45.5, 48.4, 48.6, 56.8, 112.7 (d, J=3.4 Hz), 116.7 (d, J=3.4 Hz), 119.2, 124.1 (q, J=272.9 Hz), 128.7, 128.9, 129.7, 129.8, 130.9, 131.5 (q, J=32.2 Hz), 132.8, 150.8, 165.1, 176.4. Enantiomeric purity >99% (t.sub.R=39.97 min).

Example 47. (S)-1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl) pyrrolidine-2,5-dione ((S)-6)

[0201] The compound was prepared according to procedure described in Example 44. (S)-4-oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)amino)-butanoic acid (0.93 g, 2.0 mmol, 1 eq) was used as the substrate for the cyclization reaction. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0202] White solid. Yield: 78% (0.69 g); m.p. 188.9-190.5° C.; TLC: R.sub.f=0.36 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.22F.sub.3N.sub.3O.sub.3 (445.44), Monoisotopic mass: 445.16. UPLC (purity: >99.9%): t.sub.R=6.94 min, (M+H).sup.+446.2. .sup.1H NMR (300 MHz, CDCl.sub.3) δ 2.56-2.83 (m, 5H), 3.00-3.20 (m, 2H), 3.23-3.43 (m, 3H), 3.62-3.76 (m, 1H), 3.94-4.08 (m, 1H), 6.12 (s, 1H), 6.89-6.99 (m, 2H), 7.10 (d, 1H, J=7.7 Hz), 7.28-7.53 (m, 6H); .sup.13C NMR (75 MHz, CDCl.sub.3) δ 28.0, 42.2, 45.5, 48.4, 48.6, 56.8, 112.7 (d, J=4.6 Hz), 116.7 (d, J=4.6 Hz), 124.1 (q, J=272.9 Hz), 128.7, 129.0, 129.7, 129.8, 131.6 (q, J=32.2 Hz), 132.8, 150.8, 165.1, 176.3. Enantiomeric purity >99% (t.sub.R=26.21 min).

Example 48. (R)-1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione ((R)-10)

[0203] The compound was prepared according to procedure described in Example 44. (R)-4-oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)amino)butanoic acid (0.96 g, 2.0 mmol, 1 eq) was used as starting material for the cyclization reaction. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0204] White solid. Yield: 77% (0.70 g); m.p. 168.2-169.1° C.; TLC: R.sub.f=0.46 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.22F.sub.3N.sub.3O.sub.4 (461.44), Monoisotopic mass: 461.16. UPLC (purity: >99.9%): t.sub.R=7.18 min, (M+H).sup.+462.1. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.60-2.78 (m, 5H), 2.98-3.16 (m, 2H), 3.23-3.38 (m, 3H), 3.63-3.72 (m, 1H), 3.98 (ddd, J=12.89, 6.01, 2.86 Hz, 1H), 6.11 (s, 1H), 6.61 (s, 1H), 6.69-6.73 (m, 2H), 7.21 (t, J=8.0 Hz, 1H), 7.32-7.38 (m, 3H), 7.42-7.44 (m, 2H). .sup.13C NMR (126 MHz, CDCl.sub.3) δ 28.1, 42.3, 45.6, 48.4, 48.6, 56.9, 108.9, 112.2, 114.3, 120.5 (q, J=256.7 Hz), 129.4 (d, J=143.7 Hz), 129.6 (d, J=151.5 Hz), 132.9, 150.3, 152.0, 165.2, 176.4. Enantiomeric purity >99% (t.sub.R=35.08 min).

Example 49. (R)-1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl(sulfanyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione ((R)-12)

[0205] The compound was prepared according to procedure described in Example 44. (R)-4-oxo-4-((2-oxo-1-phenyl-2-(4-(3-((trifluoromethyl)thio)phenyl)piperazin-1-yl)ethyl)amino)butanoic acid (0.99 g, 2.0 mmol, 1 eq) was used as the starting material for the cyclization reaction. The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system.

[0206] White solid. Yield: 86% (0.82 g); m.p. 155.1-155.8° C.; TLC: R.sub.f=0.48 (DCM:MeOH (9:0.5; v/v)); C.sub.23H.sub.22F.sub.3N.sub.3O.sub.3S (477.50), Monoisotopic mass: 477.13. UPLC (purity: >99.9%): t.sub.R=7.54 min, (M+H).sup.+478.1. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.68-2.75 (m, 5H), 2.96-3.19 (m, 2H), 3.22-3.43 (m, 3H), 3.62-3.76 (m, 1H), 3.99 (ddd, J=13.17, 5.73, 2.8 Hz, 1H), 6.11 (s, 1H), 6.91 (dd, J=8.3, 2.6 Hz, 1H), 7.06 (s, 1H), 7.12 (d, J=8.0 Hz, 1H), 7.24-7.28 (m, 1H), 7.33-7.38 (m, 3H), 7.42-7.44 (m, 2H). .sup.13C NMR (126 MHz, CDCl.sub.3) δ 28.1, 42.3, 45.6, 48.4, 48.7, 56.9, 123.7, 125.3, 129.6 (q, J=307, 8 Hz), 127.8, 129.4 (d, J=142.4 Hz), 129.1, 130.1, 132.9, 151.4, 165.2, 176.4. Enantiomeric purity >99% (t.sub.R=34.82 min).

Example 50. Special Properties of Enantiomers

[0207] The effect of the stereochemistry of the compounds according to invention on their anticonvulsant activity was investigated. The anticonvulsant properties were assessed in line with methods described above and results are summarized in Table 3 and Table 4.

TABLE-US-00004 TABLE 3 Data from screening studies at a dose of 100 mg/kg for selected enantiomers of compounds according to the general formula (II). Test* Compound MES 6 Hz (32 mA) 6 Hz (44 mA) scPTZ (R)-3 4/4 4/4 — 3/4 (R)-4 4/4 4/4 — 2/4 (R)-6 4/4 4/4 3/4 3/4 (S)-6 3/4 2/4 1/4 3/4 (R)-10 4/4 4/4 — 2/4 (R)-12 4/4 4/4 — 1/4 *Tests carried out in mice after intraperitoneal administration at a time point of 0.5 h, data indicate the number of mice protected in a given seizure model/number of mice tested; MES—the maximal electroshock test, the 6 Hz (32 mA) and 6 Hz (44 mA) test—the psychomotor seizures induced by low-frequency current (6 Hz) and intensity of 32 mA or 44 mA, respectively; scPTZ—the subcutaneous seizure test; “—”—substance not tested.

TABLE-US-00005 TABLE 4 The ED.sub.50 and TD.sub.50 values for selected enantiomers of compounds according to the general formula (II) and the model AED—valproic acid (VPA) after intraperitoneal administration to mice. ED.sub.50 [mg/kg] 6 Hz 6 Hz TD.sub.50 PI Compound MES (32 mA) (44 mA) scPTZ (mg/kg) (TD.sub.50/ED.sub.50) (R)-3 57.7 49.3 — 78.3 223.0 3.7 (MES) 4.5 (6 Hz, 32 mA) (R)-6 36.0 39.1 115.0 54.8 468.5 2.8 (scPTZ) 13.1 (MES) 12.0 (6 Hz, 32 mA) (S)-6 68.5 >130 — 75.4 >300 4.1 (6 Hz, 44 mA) 8.5 (scPTZ) >4.4 (MES) (R)-10 22.6 12.8 — <60.0 >150 >4.0 (scPTZ) >6.6 (MES) 11.7 (6 Hz, 32 mA) VPA.sup.g 252.7 130.6 183.1 239.4 430.7 <2.5 (scPTZ) 1.7 (MES) 3.3 (6 Hz, 32 mA) 2.3 (6 Hz, 44 mA) 1.8 (scPTZ) The substances were tested 0.5 h after intraperitoneal administration; MES—the maximal electroshock test, the 6 Hz (32 mA) and 6 Hz (44 mA) test—the psychomotor seizures induced by low-frequency current (6 Hz) and intensity of 32 mA or 44 mA, respectively; scPTZ—the subcutaneous seizure test; TD.sub.50 values were obtained in the rotarod test; PI—protective index (TD.sub.50/ED.sub.50); “—”—substance not tested.

[0208] On the basis of results obtained, it was unexpectedly found that the R-enantiomers exhibit higher biological activity with the desired profile compared to 5-enantiomers.

[0209] In particular, in the case of R enantiomers, it was found: [0210] weaker acute neurotoxicity in the rotarod test (see TD.sub.50 values in Table 2 and Table 4, respectively) in relation to the racemate, [0211] it was also unexpectedly found that the anticonvulsant effect was stereospecific. Enantiomers with the R configuration are characterized by stronger biological activity.

[0212] Metabolic stability. The metabolic stability of (R)-6 was assessed according to the methodology described above. Based on the data obtained, an extremely low value of the internal clearance of the compound (R)-6 after incubation with HLMs was found, amounting to CL.sub.int=2.4 mL/min/kg, indicating its predicted high stability in the human body. In addition, surprisingly and preferably the value of the determined clearance was lower than the value determined for the racemate, compound 6 (CL.sub.int=5.6), which indicates a lower susceptibility of the enantiomer to metabolic changes. In addition, the results of UPLC analysis revealed that the (R)-6 enantiomer is preferably metabolized to two metabolites-M1 metabolite formed by the dehydrogenation of the piperazine ring, and the M2 metabolite formed by hydroxylation of the phenyl substituent linked to piperazine (FIG. 13). In the case of the racemate, an additional M3 metabolite was observed, most likely obtained by hydroxylation of the side phenyl moiety and reduction of the keto group to hydroxyl in the imide ring (FIG. 7).

Example 51. Preparation of Water-Soluble Salts of Compounds According to the Invention

[0213] The water-soluble salts of compound according to formula (I) of the invention can be obtained applying the six-step procedure using commercially available tert-butoxycarbonyl (Boc) amino acid derivatives as starting materials. Water-soluble salts were obtained for selected compounds described by formula (I) for which k=0, D is a substituent selected from the group comprising of: H, amino group (—NH.sub.2), amino group substituted with one or two aliphatic substituents (in particular —CH.sub.3 and/or —C.sub.2H.sub.5) or an amino group that is part of a heterocyclic ring, where A and B have the meaning as in the case of compound described by formula (II).

[0214] A general scheme for the synthesis of water-soluble salts of the compounds described by formula (1) according to invention is shown in FIG. 14. In case the preparation of compounds of formula (I), where D is hydrogen, the procedure described for compounds of formula (II) according to FIG. 2B is used, after which the obtained compound is converted into a water-soluble salt (preferably a hydrochloride) using methods described in the literature.

[0215] Steps i and ii are analogous to the procedure described for the synthesis of enantiomers. The amine derivative (VI) undergoes condensation reaction with maleic anhydride, to give the compound with the unsaturated amido-acid structure (VIII). Next, compound VIII is cyclized to compound IX. In the next step, compound of formula IX is subjected to the addition reaction with the appropriate primary or secondary amine. Then the desired compound according to formula (I) is converted into a water-soluble salt (preferably a hydrochloride) using methods described in the literature.

[0216] Examples of synthesis as well as physicochemical and spectral data for selected intermediates (VIII, IX) and final products according to FIG. 14 are described below.

Example 52. 4-Oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl) amino)but-2-enoic Acid (VIII)

[0217] Maleic anhydride (0.98 g 10.0 mmol, 1 eq) was added to a solution of 2-amino-2-phenyl-1-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethan-1-one (4.61 g 10.0 mmol, 1 eq) in AcOEt (50 mL) and stirred for 30 minutes. After this time, the solvent was distilled off to dryness.

[0218] The compound was obtained as solid after washing with Et.sub.2O.

[0219] White solid. Yield: 85% (3.76 g); C.sub.23H.sub.22F.sub.3N.sub.3O.sub.4 (461.44), Monoisotopic mass: 461.16. UPLC (purity=96%): t.sub.R=6.94 min. (M+H).sup.+462.2.

Example 53. 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)-1H-pyrrole-2,5-dione (IX)

[0220] ZnCl.sub.2 (1.36 g, 10.0 mmol, 1 eq) was added to the suspension of 4-oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)amino)but-2-enoic acid (4.40 g, 10.0 mmol, 1 eq) in dry benzene (100 mL), and the mixture was heated to 80° C. Then solution of HMDS (2.42 g, 3.14 mL, 15.0 mmol, 1.5 eq) in dry benzene (10 mL) was added dropwise over 30 minutes. The reaction was continued with stirring in reflux for about 24 hours, then cooled and concentrated under reduced pressure. After distilling off the solvent, the oily residue was dissolved in DCM and extracted with 0.1 M HCl (3×50 mL), water (3×50 mL) and saturated NaCl solution (3×50 mL). The organic layer was dried over anhydrous Na.sub.2SO.sub.4 and then evaporated to dryness. The crude product was purified by column chromatography with DCM:MeOH (9:0.3; v/v) mixture as eluent system. The compound was obtained as solid after washing with Et.sub.2O.

[0221] White solid. Yield: 79% (3.34 g); C.sub.23H.sub.22F.sub.3N.sub.3O.sub.4 (443.43), Monoisotopic mass: 443.15. UPLC (purity=99%): t.sub.R=7.45 min. (M+H).sup.+444.1.

Example 54. 3-(Methylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride

[0222] 2M methylamine solution in THF (0.07 g, 2.2 mmol, 1 eq) was added to a solution of 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)-1H-pyrrole-2,5-dione (0.98 g, 2.2 mmol, 1 eq) in dry benzene (50 mL). The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system. The compound was then converted into the hydrochloride salt by treating the compound with a 2M methanolic hydrochloric acid solution.

[0223] White solid. Yield: 87% (0.91 g); m.p. 161.2-163.4° C.; C.sub.24H.sub.25F.sub.3N.sub.4O.sub.3 (474.48), Monoisotopic mass: 474.19. UPLC (purity: >99.9%): t.sub.R=5.53 min, (M+H).sup.+475.3. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.76 (br s, 3H), 2.90 (br s, 1H), 3.22 (br s, 2H), 3.38-3.54 (m, 4H), 3.55-3.66 (m, 1H), 3.70 (br s, 1H), 3.84-4.23 (m, 2H), 4.53 (br s, 1H), 6.20 (br s, 1H), 7.18-7.24 (m, 3H), 7.29-7.51 (m, 5H), 7.71 (br s, 1H), 9.98 (br s, 1H).

Example 55. 3-(Dimethylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)-piperazin-1-yl)ethyl)pyrrolidine-2,5-dione Hydrochloride

[0224] The compound was prepared according to procedure described in Example 54. 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)-1H-pyrrolo-2,5-dione (0.98 g, 2.2 mmol, 1 eq) and dimethylamine (0.10 g, 2.2 mmol, 1 eq) were used as starting materials.

[0225] The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system. The compound was converted into the hydrochloride salt by treating the compound with a 2M methanolic hydrochloric acid solution.

[0226] White solid. Yield: 83% (0.90 g); m.p. 157.8-159.2° C.; C.sub.25H.sub.27F.sub.3N.sub.4O.sub.3 (488.51), Monoisotopic mass: 488.20. UPLC (purity: >99.9%): t.sub.R=5.53 min, (M+H).sup.+489.3. .sup.1H NMR (500 MHz, CDCl.sub.3) δ 2.76 (d, J=8.6 Hz, 1H), 2.93 (br s, 2H), 3.06-3.18 (m, 5H), 3.25-3.33 (m, 3H), 3.36-3.41 (m, 2H), 3.41-3.45 (m, 2H), 3.71 (br s, 1H), 3.94-3.98 (m, 1H), 6.14 (s, 1H), 7.01 (d, J=7.4 Hz, 1H), 7.04 (br s, 1H), 7.12 (d, J=7.4 Hz, 1H), 7.34 (t, J=7.7 Hz, 1H), 7.39 (s, 5H), 13.02 (br s, 1H). .sup.13C NMR (126 MHz, CDCl.sub.3) δ 31.4, 42.5 45.7, 48.7, 48.9, 57.7, 60.1, 65.9, 113.1, 117.5, 119, 3, 119.9, 119.7, 124.1 (d, J=272.2 Hz) 129.1, 129.8, 129.9, 131.1, 131.7 (d, J=32.0 Hz) 150.5, 164.4, 169.8, 171.7.

Example 56. 3-(Diethylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione Hydrochloride

[0227] The compound was prepared according to procedure described in Example 54. 1-(2-Oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)-1H-pyrrolo-2,5-dione (0.98 g, 2.2 mmol, 1 eq) and diethylamine (0.16 g, 2.2 mmol, 1 eq) were used as starting materials.

[0228] The crude product was purified by column chromatography using DCM:MeOH (9:0.5; v/v) eluent system. The compound was converted into the hydrochloride salt by treating the compound with a 2M methanolic hydrochloric acid solution.

[0229] White solid. Yield: 88% (1.00 g); m.p. 142.2-143.1° C.; TLC: R.sub.f=0.52 (DCM:MeOH (9:0.5; v/v)); C.sub.27H.sub.31F.sub.3N.sub.4O.sub.3 (516.57), Monoisotopic mass: 516.23. UPLC (purity: >99.9%): t.sub.R=5.79 min, (M+H).sup.+517.2. .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 1.17-1.27 (m, 6H), 2.79-2.89 (m, 1H), 3.05-3.36 (m, 9H), 3.54-3.78 (m, 3H), 4.79 (dd, J=9.2, 5.7 Hz, 1H), 4.92 (dd, J=9.2, 5.7 Hz, 1H), 6.20 (s, 1H), 7.04 (d, J=7.4 Hz, 1H), 7.10 (s, 2H), 7.14 (d, J=8.0 Hz, 1H), 7.31-7.37 (m, 5H), 12.88 (br s, 1H).

Example 57. Special Properties of the Water-Soluble Salts of Compounds According to the Invention

[0230] The effect of improved water solubility (i.e. salts) of compounds according to the invention on their anticonvulsant activity was investigated. The anticonvulsant properties were assessed in line with methods described above and results are summarized in Table 3 and Table 4.

TABLE-US-00006 TABLE 5 Data from screening studies at a dose of 100 mg/kg for selected water-soluble salts according to the general formula (I). Test* Compound MES 6 Hz (32 mA) 6 Hz (44 mA) scPTZ 48 4/4 4/4 — 3/4 49 4/4 4/4 — 3/4 50 4/4 4/4 — 2/4 *Tests carried out in mice after intraperitoneal administration at a time point of 0.5 h, data indicate the number of mice protected in a given seizure model/number of mice tested; MES—the maximal electroshock test, the 6 Hz (32 mA) and 6 Hz (44 mA) test—the psychomotor seizures induced by low-frequency current (6 Hz) and intensity of 32 mA or 44 mA, respectively; scPTZ—the subcutaneous seizure test; “—”—substance not tested.

TABLE-US-00007 TABLE 6 The ED.sub.50 and TD.sub.50 values for selected water-soluble salt of compounds according to the general formula (I) and the model AED—valproic acid (VPA) after intraperitoneal administration to mice. ED.sub.50 [mg/kg] 6 Hz 6 Hz TD.sub.50 PI Compound MES (32 mA) (44 mA) scPTZ (mg/kg)b (TD.sub.50/ED.sub.50) 23 77.5 80.4 — <100 246.6 3.2 (MES) 3.0 (6 Hz, 32 mA) >2.5 (scPTZ) VPA 252.7 130.6 183.1 239.4 430.7 1.7 (MES) 3.3 (6 Hz, 32 mA) 2.3 (6 Hz, 44 mA) 1.8 (scPTZ) The substances were tested 0.5 h after intraperitoneal administration; MES—the maximal electroshock test, the 6 Hz (32 mA) and 6 Hz (44 mA) test—the psychomotor seizures induced by low-frequency current (6 Hz) and intensity of 32 mA or 44 mA, respectively; scPTZ—the subcutaneous seizure test; TD.sub.50 values were obtained in the rotarod test; PI—protective index (TD.sub.50/ED.sub.50); “—”—substance not tested.

[0231] Based on the results obtained, it was found that the salts of the compounds according to the invention show distinctly improved water solubility. This positively affects their pharmacokinetics or/and pharmaceutical properties, and is particularly advantageous in case of intravenous administration of compounds according to the invention.