(2,5-dioxopyrrolidin-1-yl)(phenyl)-acetamide derivatives and their use in treatment of neurological diseases

Abstract

To provide compounds for use in the treatment or prevention of epileptic seizures or neuropathic pain or migraine.SOLUTION: The invention provides the compound of general formula (I) or pharmaceutically acceptable salts thereof. The invention also provides the use of compounds described by general formula (I) as active ingredients in pharmaceutical compositions for the treatment of epileptic seizures or neuropathic pain or migraine.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to (2,5-dioxopyrrolidin-1-yl)(phenyl)-acetamide derivatives and their pharmaceutically acceptable salts, suitable for the treatment of neurological disorders. The disclosed compounds exhibit a wide range of protective activity in animal models of epileptic seizures and pain models, and therefore can find application in the treatment of neurological disorders, particularly epilepsy and neuropathic pain. Due to the wide range of therapeutic applications of antiepileptic drugs, these compounds can also be useful in the treatment of, for example, migraine, withdrawal syndrome, schizophrenia, schizoaffective disorder, personality and nutritional disorders, and anxiety and post-traumatic stress. [Background technology]

[0002] Epilepsy is one of the most common neurological disorders associated with impaired excitability and neurotransmission. It affects 1-2% of the human population and significantly reduces patients' quality of life and ability to perform daily activities (Nadkarni, S.; LaJoie, J.; Devinsky, O., Neurology 2005, 64, S2-S11). Due to its complex pathophysiology, epilepsy is a heterogeneous disorder characterized by the development of various seizure types (e.g., tonic-clonic, absence, partial, etc.) and significant drug resistance, which reaches 30-40% of diagnosed cases (Kwan, P.; Schachter, SC; Brodie, MJ, N. Engl. J. Med. 2011, 365, 919-926). Neuropathic pain is another severe neurological disorder that presents therapeutic challenges. Current data indicate that only 50% of patients achieve a 30-50% reduction in neuropathic pain sensation, while others fail to achieve improvement with any of the drugs used (Butera, JA, J. Med. Chem. 2007, 11, 2543-2546). Therefore, there is a great need for novel AEDs that can control various types of epileptic seizures, preferably those that are effective against neuropathic pain. Many currently used AEDs have narrow therapeutic indications, meaning they are only applicable to specific types of epileptic seizures. These drugs include newer AEDs such as levetiracetam and lacosamide, among others. Recent research has shown that multitarget compounds, also known as multifunctional compounds, i.e., compounds with complex molecular mechanisms of action, may be particularly beneficial for the treatment of diseases with complex pathological mechanisms (so-called multifactorial diseases). By combining different synergistic mechanisms, multi-targeting substances are believed to ensure higher therapeutic efficacy compared to substances acting on a single biological target, as they enable a comprehensive therapeutic process (Bansal, Y.; Silakari, O., Eur. J. Med. Chem. 2014, 76, 31-42).Another advantage of multifunctional drugs may be a reduction in the number of drugs taken, which may result in fewer and less severe side effects, a lower risk of drug-drug interactions, and better cooperation (compliance) between doctors and patients. It is also hypothesized that multitarget compounds may be useful in treating diseases characterized by high drug resistance (e.g., epilepsy) (Talevi, A., Front. Pharmacol. 2015, 6, 205). Multitarget substances are usually designed as hybrid or chimeric molecules that combine structural fragments of a common chemical framework responsible for specific pharmacological effects (Morphy, R.; Rankovic, Z., J. Med. Chem. 2005, 48, 6523-6543). In particular, intensive research into the development of multitarget compounds as candidates for new drugs is being conducted in the fields of cancer, neurodegenerative, and inflammatory diseases.In particular, the concept of molecular hybridization has recently been proposed by the present inventors as a method to enable the design and development of novel AEDs with a wide range of therapeutic applications (Abram, M.; Zagaja, M.; Mogilski, S.; Andres-Mach, M.; Latacz, G.; Bas, S.; Luszczki, JJ; Kiec-Kononowicz, K.; Kaminski, K., J. Med. Chem. 2017, 60, 8565-8579; Kaminski, K.; Zagaja, M.; Rapacz, A.; Luszczki, JJ; Andres-Mach, M.; Abram, M.; Obniska, J., Bioorg. Med. Chem. 2016, 24, 606-61). 8 pages; Kaminski, K.; Rapacz, A.; Filipek, B.; Obniska, J., Bioorg. Med. Chem. 2016, 24, 2938~2946 Page; Kaminski, K.; Zagaja, M.; Luszczki, JJ; Rapacz, A.; Andres-Mach, M.; Latacz, G.; Kiec-Konon owicz, K., J. Med. Chem. 2015, 58, pp. 5274-5286; Kaminski, K.; Rapacz, A.; Luszczki, JJ; Latacz, G.; Obniska, J.; Kiec-Kononowicz, K.; Filipek, B., Bioorg. Med. Chem. 2015, 23, pp. 2548-2561). Summary of the Invention [Problem to be solved by the invention]

[0003] The anticonvulsant and / or analgesic activity of novel compounds is routinely evaluated in animal models (primarily mice and rats). From a clinical perspective, particularly promising candidates for novel broad-spectrum AEDs effective against various types of human epileptic seizures are those active in the maximal electroshock test (MES), the subcutaneous pentylenetetrazol seizure test (scPTZ), and the psychomotor 6-Hz seizure model (at current intensities of 32 mA or / and 44 mA) utilizing a 6-Hz low-frequency current. Compounds with such profiles in preclinical in vivo studies may be effective against human tonic-clonic seizures with or without secondary generalization, generalized absence seizures, myoclonic seizures, partial seizures, and drug-resistant epilepsy. An important added value of these compounds should be their activity in key animal tests / models assessing antinociceptive activity, namely, the formalin test, the capsaicin-induced pain model, and the oxaliplatin-induced neuropathic pain model.

[0004] The technical problem before the present invention was to provide such chemical compounds, or pharmaceutically acceptable salts thereof, which are easily obtainable, do not show hepatotoxic effects, and which can be used as active substances in pharmaceutical compositions to control various types of seizures (secondarily generalized, generalized absence seizures, myoclonic seizures, partial seizures, and tonic-clonic seizures with or without drug-resistant epilepsy), and which should also have analgesic activity in pain caused by neuropathic disorders or migraine. [Means for solving the problem]

[0005] A first subject of the present invention is a compound of general formula (I)

[0006] [ka] (In the formula, X is N or C; k is a number equal to 0 or 1, A is, - phenyl substituents; - phenyl substituents substituted with one or two or three or four lateral substituents selected from the group consisting of halogen atoms, -SCF3, -CF3, -CHF2, -CN, -OCF3, -NO2, -OCH3, -OC2H5, alkyl moieties having 1 to 4 carbon atoms in the carbon backbone, the alkyl moiety having a linear or branched chain; - a phenyl substituent substituted with at least one aromatic or heteroaromatic substituent; - benzhydryl substituents; - 1-naphthyl or 2-naphthyl substituents; - a benzothiophenyl substituent selected from the group consisting of a 2-benzothiophenyl, a 3-benzothiophenyl, a 4-benzothiophenyl, or a 5-benzothiophenyl substituent, preferably a 5-benzothiophenyl substituent; - a benzisoxazole substituent selected from the group consisting of 3-benzisoxazole, 4-benzisoxazole, 5-benzisoxazole, 6-benzisoxazole, 7-benzisoxazole substituents, preferably a 5-benzisoxazole substituent; - an alkyl moiety having 1 to 4 carbon atoms in the carbon backbone, which has a linear, branched or cyclic chain, and which is preferably substituted with at least one halogen atom; is a substituent selected from the group consisting of: B is - phenyl substituents; - phenyl substituents substituted with one or two lateral substituents selected from the group consisting of halogen atoms, -SCF3, -CF3, -CHF2, -CN, -OCF3, -NO2, -OCH3, -OC2H5, alkyl moieties having 1 to 4 carbon atoms in the carbon backbone, the alkyl moieties having a linear or branched chain; Defined as; D is a substituent selected from the group consisting of H, amino (-NH), an amino group substituted with one or two aliphatic substituents (including, in particular, -CH and / or -C H), or an amino group that is part of a heterocycle. or a pharmaceutically acceptable salt thereof.

[0007] The term "halogen", when used to describe compounds according to general formula (I), includes fluorine, chlorine, bromine, and iodine. In another preferred embodiment of the present invention, the halogen atom is fluorine or chlorine.

[0008] The compound having the general formula (I) has multiple chiral centers, and therefore can exist in the form of optical isomers and their mixtures.Various ratios of the aforementioned optical isomers and their mixtures, including racemic mixtures, are included within the scope of the present invention.Individual isomers can be obtained by using the appropriate isomeric form of the starting material (amino acid derivative), or can be separated after preparing the final compound according to known separation methods.

[0009] Preferably, the compounds of the present invention have the general formula (II)

[0010] [ka] (In the formula, X is N or C; k is a number equal to 0 or 1, A is, - phenyl substituents; - phenyl substituents substituted with one or two or three or four lateral substituents selected from the group consisting of halogen atoms, -SCF3, -CF3, -CHF2, -CN, -OCF3, -NO2, -OCH3, -OC2H5, alkyl moieties having 1 to 4 carbon atoms in the carbon backbone, the alkyl moiety having a linear or branched chain; - a phenyl substituent substituted with at least one aromatic or heteroaromatic substituent; - benzhydryl substituents; - 1-naphthyl or 2-naphthyl substituents; - a benzothiophenyl substituent selected from the group consisting of a 2-benzothiophenyl, a 3-benzothiophenyl, a 4-benzothiophenyl, or a 5-benzothiophenyl substituent, preferably a 5-benzothiophenyl substituent; - a benzisoxazole substituent selected from the group consisting of 3-benzisoxazole, 4-benzisoxazole, 5-benzisoxazole, 6-benzisoxazole, 7-benzisoxazole substituents, preferably a 5-benzisoxazole substituent; - an alkyl moiety having 1 to 4 carbon atoms in the carbon backbone, which has a linear, branched or cyclic chain, and which is preferably substituted with at least one halogen atom; is a substituent selected from the group consisting of: B is - phenyl substituents; - phenyl substituents substituted with one or two lateral substituents selected from the group consisting of halogen atoms, -SCF3, -CF3, -CHF2, -CN, -OCF3, -NO2, -OCH3, -OC2H5, alkyl moieties having 1 to 4 carbon atoms in the carbon backbone, the alkyl moieties having a linear or branched chain; is) or a pharmaceutically acceptable salt thereof. Preferably, the halogen is a fluorine or chlorine atom.

[0011] Preferably, the alkyl moieties in the carbon backbone contain 1 to 4 carbon atoms, the alkyl moieties having straight or branched chains and selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, and tert-butyl substituents.

[0012] The compound having general formula (II) has one chiral center, and therefore can exist in the form of optical isomers and their mixtures.Various ratios of the aforementioned optical isomers and their mixtures, including racemic mixtures, are included within the scope of the present invention.Individual isomers can be obtained by using the appropriate isomeric form of the starting material (amino acid derivative), or can be separated after preparing the final compound according to known separation methods.

[0013] Preferably, k=0.

[0014] Preferably, the X atom is a nitrogen atom.

[0015] Preferably, the substituent A is selected from the group consisting of 5-benzothiophenyl, 2-naphthyl, and 5-benzisoxazolyl substituents.

[0016] Preferably, A is selected from the group consisting of phenyl; phenyl substituted with at least one chlorine or -CF3, -CHF2, -OCF3, -CH3, -SCF3 or phenyl.

[0017] Preferably, the substituent B is selected from the group consisting of phenyl or phenyl substituted with one or two halogen atoms.

[0018] Preferably, the compound of the present invention is selected from the group consisting 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-yl)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.

[0019] Preferably, the compounds of the present invention are the (R) enantiomers and are 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.

[0020] Preferably, the compounds of the present invention are water-soluble salts, especially hydrochlorides, 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-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride.

[0021] A second subject of the present invention is a compound according to the invention as defined above, for use in the treatment or prevention of epileptic seizures, neuropathic pain or migraines. In a preferred embodiment, the compound according to the invention is used as the only active substance or one of many contained in a pharmaceutical composition for the treatment or prevention of at least one of the above medical indications.

[0022] The compounds according to the invention have anticonvulsant and analgesic activity in a wide panel of animal models and may find application as active substances in various dosage forms for the treatment of epilepsy and neuropathic pain.

[0023] The compounds of formula (I) according to the present invention can be obtained using a multi-step synthetic procedure, which is illustrated in Figure 2A, where X, A, B, D, and k are as defined above. For the preparation of compounds of formula (I) where D is a halogen, the procedure described for compounds of formula (II) according to Figure 2B is used. In the first step, an intermediate product with an amide structure is obtained as a result of a condensation reaction (i) between an appropriate piperazine derivative and the corresponding tert-butoxycarbonyl (Boc) amino acid derivative, which is then subjected to a deprotection reaction (ii) to form an amine derivative. In the next step, the aforementioned amine derivative is subjected to a condensation reaction (iii) with maleic anhydride to obtain an unsaturated amide-acid derivative. This derivative is then subjected to a cyclization reaction (iv) to form the corresponding maleimide. In the next step (v), the maleimide derivative is subjected to an addition reaction with a suitable primary or secondary amine to obtain a compound having the general formula (I) according to the present invention.

[0024] The compound of formula (II) according to the present invention has the formula (III):

[0025] [ka] wherein B and k are as defined for formula (II). The compound of formula (III) can be obtained by a two-step procedure using commercially available succinic anhydride and the corresponding amino acid derivative as a substrate. In the first step, the intermediate product of the amide-acid structure (IV) is obtained as a result of the condensation reaction of succinic anhydride with the appropriate amino acid derivative, which then undergoes a cyclization reaction to form the desired compound having formula (III). Alternatively, the compound described by formula (III) can be prepared by using a one-step thermal cyclocondensation reaction between succinic anhydride or succinic acid and the corresponding amino acid.

[0026] The desired compound of general formula (II) according to the present invention can be obtained by amidation reaction between the compound described by formula (III) and a suitable commercially available secondary aliphatic amine. This reaction can be carried out in the presence of known coupling agents, including CDI, EDCI, DCC, etc. Alternatively, the compound of formula (II) can be obtained by reaction between the acid chloride obtained by conversion of the carboxylic acid described by formula (II) and the corresponding commercially available secondary aliphatic amine. The compound of formula (II) according to the present invention can also be prepared by reaction between a carboxylic acid and the corresponding aliphatic amine in the presence of an organic base, particularly triethylamine (TEA), N-methylmorpholine (NMM), or N,N-diisopropylethylamine (DIEA), using an activating agent selected from BOP, HBTU, and HATU.

[0027] The synthetic procedure and reaction conditions are illustrated in Figure 2B, where X, A, B, and k are as defined above.

[0028] The solution according to the present invention has several advantages. The disclosed compounds of formula (I), preferably compounds of formula (II), are characterized by strong and broad anticonvulsant activity in various animal models of epilepsy, i.e., the maximal electroshock seizure test (MES), the subcutaneous pentylenetrazole seizure test (scPTZ), and the 6 Hz seizure model (32 mA or / and 44 mA). Compounds with the above profile in preclinical in vivo studies may be effective in various types of human epilepsy, including secondarily generalized, generalized (absence), myoclonic, partial seizures, and tonic-clonic seizures with or without significant drug-resistant seizures. Another advantage of the compounds of general formula (I), especially those described by formula (II), is their strong analgesic activity in animal models assessing antinociceptive activity, i.e., the formalin test, the capsaicin-induced pain model, and the oxaliplatin-induced neuropathic pain model. Therefore, the compound of formula (I), preferably the compound of formula (II), can be useful for treating both neuropathic and inflammatory pain, which is a unique feature among the AEDs available for pharmacological therapy.The compound of formula (I), preferably the compound of formula (II), has a complex molecular mechanism of action, namely, they interact with voltage-dependent sodium channel, calcium channel and TRPV1 receptor.The beneficial antagonistic effect observed in the case of TRPV1 receptor has not yet been demonstrated for known and therapeutically relevant AEDs. Importantly, literature data suggest a possible involvement of TRPV1 in seizure induction (Naziroglu, M., Curr. Neuropharmacol. 2015, 13, 239-247; Naziroglu, M.; Ovey, IS, Neuroscience 2015, 293, 55-66), while its role as a molecular target for substances with antinociceptive activity is well documented (Szallasi, A.; Cortright, DN; Blum, CA; Eid, SR, Nat. Rev. Drug. Discov. 2007, 6, 357-372).The compound of formula (II) can also be potentially useful in the treatment of, among others, withdrawal syndrome, schizophrenia, schizoaffective disorder, personality and nutritional disorders, and anxiety and post-traumatic stress.Therefore, the present invention provides compounds for use as drugs.Furthermore, the present invention discloses the possibility of using TRPV1 receptor antagonists to treat various types of epileptic seizures.

[0029] The compounds of the present invention can be administered by various routes, including enteral, topical, or parenteral administration, containing at least one active compound according to Formula (I), preferably Formula (II), in a pharmaceutically acceptable and effective amount, along with pharmaceutically acceptable diluents, carriers, and / or excipients known in the art, to form pharmaceutical preparations suitable for a given route of administration. Methodologies for preparing such pharmaceutical formulations are known in the art. Therapeutic doses vary depending on the substance, species, sex, age, disease entity being treated, route, and method of administration, and must be determined by those skilled in the art. The suggested dose of the compounds of the present invention is 0.1 to about 1000 mg per day, in single or divided doses. The compounds of the present invention can be administered to a patient either by themselves or in combination with one or more active ingredients, each present in its own composition, or some or all of the active ingredients combined in a single composition, and / or suitable pharmaceutical excipients. Suitable pharmaceutical excipients include conventional supporting substances necessary for the proper preparation of a given formulation, such as fillers, binders, disintegrants, lubricants, solvents, gel-forming agents, emulsifiers, stabilizers, dyes, and / or preservatives. The compounds of the present invention are formulated into dosage forms using commonly known pharmaceutical preparation methods. Dosage forms can be, for example, tablets, capsules, granules, suppositories, emulsions, suspensions, or solutions. Depending on the method of administration and galenic form, the amount of active substance in the formulation can typically range from 0.01% (by weight) to 100% (by weight).

[0030] Embodiments of the present invention are illustrated in the drawings shown below. [Brief explanation of the drawings]

[0031] [Figure 1] FIG. 1 shows the general formula of compounds (I) and (II). [Figure 2A] FIG. 1 shows the synthesis of derivatives according to formula (I). [Figure 2B] FIG. 1 illustrates the synthesis of derivatives according to formula (II). [Figure 3] 1 shows the analgesic activity of Compound 6 and valproic acid (VPA) in Phase I and Phase II pain of the formalin test. Results are presented as paw licking times during Phase I (0-5 minutes after formalin injection) and Phase II (15-30 minutes after formalin injection) of the test. Values represent the mean ± SEM for groups of 8-10 animals; statistically significant differences compared 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. [Figure 4] 1 is a graph showing the analgesic activity of Compound 6 and valproic acid (VPA) in the capsaicin assay. Results are shown as the paw licking time 0-5 minutes after capsaicin injection. Values represent mean ± SEM; statistically significant differences compared 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. [Figure 5]Graphs showing the analgesic activity of Compound 6 and valproic acid (VPA) in oxaliplatin-induced peripheral neuropathy. A: Effect of Compound 6 on mechanical allodynia in the von Frey test. B: Effect of valproic acid (VPA) on mechanical allodynia in the von Frey test. C: Effect of Compound 6 on thermal allodynia in the cold plate test. Results are presented as mean ± SEM for groups of 8-10 animals, either paw pressure (von Frey test) or latency to onset of the nociceptive cold plate response; statistically significant differences compared to the control group (mice administered OXPT and before test compound). 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). [Figure 6] Graphs showing the effect of Compound 6 (at a concentration of 100 μM) on fast voltage-dependent sodium currents. A: Example traces of maximum voltage-dependent sodium currents in the control, in the presence of Compound 6, and after washout of Compound 6. B: Normalized mean current amplitudes in the control, in the presence of Compound 6 (*p<0.001, ANOVA with Tukey's test), and after washout of Compound 6. I / Imax (on the vertical axis) means that the current is normalized to the control value. [Figure 7] 1 is a graph showing UPLC analysis of the metabolism of compound 6 after incubation with HML. [Figure 8] 1 is a graph showing the effect of verapamil, Na3VO4, and Compound 6 on baseline Pgp activity. Statistical significance was calculated by one-way ANOVA analysis of variance and Bonferroni method (**p<0.01, ***p<0.001, compounds tested in triplicate). [Figure 9]A: Graph showing the effect of the reference inhibitor ketoconazole (KE) and compound 6 on CYP3A4 activity. B: Graph showing the effect of the reference inhibitor quinidine (QD) and 6 on CYP2D6 activity. Statistical significance was calculated by one-way ANOVA analysis of variance and Bonferroni method (***p<0.001). [Figure 10] Graph showing the effect of the reference cytostatic agent doxorubicin (DX), the mitochondrial toxin CCCP (carbonyl cyanide m-chlorophenylhydrazone), and compound 6 on cell viability of the HepG2 line after 72 hours of incubation. Statistical significance was calculated by one-way ANOVA analysis of variance and the Bonferroni method (*p<0.05, ***p<0.001, compounds tested in quadruplicate). [Figure 11] Graph showing ATP levels in HepG2 cell line after 3 hours of incubation. Doxorubicin (DX), CCCP (carbonyl cyanide m-chlorophenylhydrazone). Statistical significance was calculated by one-way ANOVA analysis of variance and Bonferroni method (***p<0.001, compounds tested in 4 replicates). [Figure 12] FIG. 1 shows a general scheme for the synthesis of enantiomers of compounds according to formula (II). [Figure 13] 1 is a graph showing UPLC analysis of the metabolism of the (R)-6 enantiomer after incubation with HML. [Figure 14] FIG. 1 shows a general scheme for the synthesis of water-soluble salts of compounds according to formula (I). DETAILED DESCRIPTION OF THE INVENTION

[0032] Analysis method: Proton magnetic resonance ( 1 H NMR) and carbon nuclear magnetic resonance ( 13C NMR spectra were recorded using a Mercury-300 "Varian" spectrometer (Varian Inc., Palo Alto, CA, USA) operating at 300 MHz and 75 MHz, respectively, or a JEOL-500 spectrometer (JEOL USA, Inc., MA, USA) operating at 500 MHz and 126 MHz, respectively. Chemical shifts are given in δ (ppm) relative to TMS δ = 0 (1H) as the internal standard. J values are expressed in hertz (Hz). Deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-D6) was used as the solvent. The following signal abbreviations are used in the spectral descriptions: s (singlet), br s (broad singlet), d (doublet), dd (doublet of doublet), ddd (doublet of doublet of doublet), t (triplet), td (triplet of doublet), q (quartet), and m (multiplet). The UPLC / MS analytical system consisted of a Waters ACQUITY® UPLC® instrument (Waters Corporation, Milford, MA, USA) equipped with a Waters TQD mass spectrometer operated in electrospray ionization (ESI) mode. Chromatographic separation was performed using an Acquity UPLC BEH C18, 1.7 μm (2.1 × 100 mm) column with a VanGuard Acquity UPLC BEH C18, 1.7 μm (2.1 × 5 mm) column (Waters, Milford, CT, USA). The column was maintained at 40 °C and eluted with a gradient of 95% to 0% eluent A over 10 min at 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 range of 200–700 nm at a resolution of 1.2 nm, with a sampling rate of 20 points / s. UPLC retention time (t R Thin layer chromatography (TLC) was performed on silica gel 60 F using the following developing solvent system: 254The separations were carried out on aluminum sheets (Macherey-Nagel, Düren, Germany) coated with the following solvents: DCM:MeOH (9:0.2; v / v), DCM:MeOH (9:0.3; v / v), DCM:MeOH (9:0.5; v / v), and DCM:MeOH (9:1; v / v). Spot detection: UV light (λ = 254 nm). Melting points (mp) were determined using open capillary tubes on a Büchi 353 instrument (Büchi Labortechnik, Flawil, Switzerland). Enantiomeric purity was determined using chiral HPLC techniques on a Shimadzu Prominence and LC-2030C SD Plus instrument (Shimadzu Corporation, Kyoto, Japan) equipped with an Amylose-C (250 × 4.6 mm) chiral column. The analysis was carried out under the following conditions: column temperature: 20°C, eluent mixture: hexane / i-PrOH = 80 / 20 (v / v), flow rate: 1 mL / min, detection at wavelength λ = 206 nm. The enantiomeric purity is expressed in %.

[0033] The preparation of compounds of the invention is illustrated in the following examples. The syntheses presented in the examples below have not been optimized in terms of yield, amount of reagents used, or the final form of the compounds obtained.

[0034] Abbreviations used: AcOEt: ethyl acetate CDI: carbonyldiimidazole DCC: N,N'-dicyclohexylcarbodiimide DCM: dichloromethane DMF: dimethylformamide Et2O: Diethyl ether HCl: Hydrochloric acid HMDS: hexamethyldisilazane MeOH: Methanol NaCl: Sodium chloride Na2SO4: Sodium sulfate ZnCl2: Zinc chloride [Example]

[0035] Synthesis, physicochemical, and spectral data of intermediates (IV and III according to the scheme in Figure 2B ). Intermediate IV: 4-((carboxy(phenyl)methyl)amino)-4-oxobutanoic acid 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, the acetic acid was evaporated to dryness. Intermediate IV was obtained as a solid after washing with EtO.

[0036] White solid. Yield: 87% (6.55 g); mp 199.4-200.6 °C; TLC: R f = 0.25 (DCM:MeOH (9:1; v / v)); C 12 H 13 NO5 (251.24), monoisotopic mass: 251.08. UPLC (100% purity): t R =2.77 min. (M+H) + 252.1.

[0037] Intermediate III: 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid ZnCl (2.73 g, 20 mmol, 1 equiv.) was added to a suspension of 4-((carboxy(phenyl)methyl)amino)-4-oxobutanoic acid (5.0 g, 20 mmol, 1 equiv.) (IV) in dry benzene (100 mL) and heated to 80 °C. A solution of HMDS (4.84 g, 6.25 mL, 30 mmol, 1.5 equiv.) in dry benzene (15 mL) was then added dropwise over 30 min. The reaction was continued to stir under reflux for approximately 24 h and then concentrated under reduced pressure. After removal of 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 NaSO and then evaporated to dryness. 2-(2,5-Dioxopyrrolidin-1-yl)-2-phenylacetic acid was obtained as a solid material after washing with Et 2 O. Alternatively, 1,4-dioxane can be used instead of benzene in the above procedure.

[0038] White solid. Yield: 90% (4.20 g); mp 195.5-198.2 °C; TLC: R f = 0.45 (DCM:MeOH (9:1; v / v)); C 12 H 11 NO4 (233.22), monoisotopic mass: 233.07. UPLC (100% purity): R =3.41 min. (M+H) + 234.1. 1 H NMR (300 MHz, DMSO-D6) 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]

[0039] 1-(2-oxo-1-phenyl-2-(4-phenylpiperazin-1-yl)ethyl)pyrrolidine-2,5-dione Carbonyldiimidazole (1.17 g, 7.2 mmol, 1.2 equiv.) 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 equiv.) dissolved in 10 mL of anhydrous DMF. After stirring for 0.5 h, a solution of 1-phenylpiperazine (0.97 g, 6 mmol, 1 equiv.) in 5 mL of anhydrous DMF was added dropwise. The reaction was allowed to stir at room temperature for 24 h. After this time, the DMF was evaporated under reduced pressure. The crude product was purified by column chromatography using a mixture of DCM:MeOH (9:0.3; v / v) as the solvent system. The compound was obtained as a solid after washing with EtO.

[0040] White solid. Yield: 84% (1.90 g); mp 156.7-157.4 °C; TLC: R f = 0.35 (DCM:MeOH (9:0.3; v / v)); C 22 H 23 N3O3 (377.44), monoisotopic mass: 377.17. UPLC (100% purity): t R=5.88 minutes. (M+H) + 378.1. 1 H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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]

[0041] 1-(2-(4-(3-chlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-(3-chlorophenyl)piperazine (1.40 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.2; v / v) elution system.

[0042] White solid. Yield: 81% (2.00 g); mp 128.1-129°C; TLC: R f = 0.51 (DCM:MeOH (9:0.2; v / v)); C 22 H 22 ClN3O3 (411.89), monoisotopic mass: 411.13. UPLC (100% purity): R = 6.69 min, (M+H) + 412.1. 1H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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]

[0043] 1-(2-(4-(3,5-dichlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-(3,5-dichlorophenyl)piperazine (1.20 g, 6 mmol, 1 equivalent). The crude product was purified by column chromatography using a DCM:MeOH (9:0.3; v / v) eluent.

[0044] White solid. Yield: 77% (2.06 g); mp 163.8-165.2 °C; TLC: R f = 0.42 (DCM:MeOH (9:0.2; v / v)); C 22 H 21 Cl2N3O3 (446.33), monoisotopic mass: 446.10. UPLC (99% purity): t R = 7.59 min, (M+H) + 446.1. 1H NMR (500 MHz, CDCl3) 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). 13 C NMR (126 MHz, CDCl3) 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]

[0045] 1-(2-oxo-1-phenyl-2-(4-(m-tolyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-(3-methylphenyl)piperazine (1.18 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.3; v / v) elution system.

[0046] White solid. Yield: 86% (2.02 g); mp 188.7-192.1 °C; TLC: R f = 0.45 (DCM:MeOH (9:0.3; v / v)); C 23 H 25 N3O3 (391.47), monoisotopic mass: 391.19. UPLC (98.9% purity): R = 6.35 min, (M+H) + 392.2. 1H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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]

[0047] 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-[3-(trifluoromethyl)phenyl]piperazine (1.38 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.2; v / v) elution system.

[0048] White solid. Yield: 82% (2.19 g); mp 150.3-151.4 °C; TLC: R f = 0.34 (DCM:MeOH (9:0.2; v / v)); C 23 H 22 F3N3O3 (445.44), monoisotopic mass: 445.16. UPLC (100% purity): t R = 6.94 min, (M+H) + 446.2. 1H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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]

[0049] 1-(2-oxo-1-phenyl-2-(4-(4-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-[4-(trifluoromethyl)phenyl]piperazine (1.38 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.3; v / v) elution system.

[0050] White solid. Yield: 62% (1.66 g); mp 173.2-174.3 °C; TLC: R f = 0.49 (DCM:MeOH (9:0.3; v / v)); C 23 H 22 F3N3O3 (445.44), monoisotopic mass: 445.16. UPLC (100% purity): t R = 6.89 min, (M+H) +446.2. 1 H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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]

[0051] 1-(2-(4-(3,5-bis(trifluoromethyl)phenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-[3,5-bis(trifluoromethyl)phenyl]piperazine (1.18 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0052] White solid. Yield: 69% (2.12 g); mp 228.1-229.4 °C; TLC: R f = 0.47 (DCM:MeOH (9:0.5; v / v)); C 24 H 21 F6N3O3 (513.44), monoisotopic mass: 513.13. UPLC (100% purity): t R = 6.58 min, (M+H) + 514.1. 1H NMR (300 MHz, CDCl3) 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]

[0053] 1-(2-oxo-1-phenyl-2-(4-(3-(difluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-(3-difluoromethylphenyl)piperazine (1.27 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.2; v / v) elution system.

[0054] White solid. Yield: 83% (2.13 g); mp 156.4-157.6 °C; TLC: R f = 0.55 (DCM:MeOH (9:0.2; v / v)); C 23 H 23 F2N3O3 (427.45), monoisotopic mass: 427.17. UPLC (100% purity): R = 6.36 minutes, (M+H) + 428.2. 1 H NMR (300 MHz, CDCl3) 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]

[0055] 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-[3-(trifluoromethoxy)phenyl]piperazine (1.48 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.3; v / v) elution system.

[0056] White solid. Yield: 89% (2.46 g); mp 100.3-101.6 °C; TLC: R f = 0.42 (DCM:MeOH (9:0.3; v / v)); C 23 H 22 F3N3O4 (461.44), monoisotopic mass: 461.16. UPLC (100% purity): t R = 7.15 min, (M+H) + 462.2. 1 H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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]

[0057] 1-(2-oxo-1-phenyl-2-(4-(4-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-[4-(trifluoromethoxy)phenyl]piperazine (1.48 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.3; v / v) elution system.

[0058] White solid. Yield: 83% (2.29 g); mp 102.3-103.5°C; TLC: R f = 0.43 (DCM:MeOH (9:0.3; v / v)); C 23 H 22 F3N3O4 (461.44), monoisotopic mass: 461.16. UPLC (100% purity): t R = 7.17 min, (M+H) + 462.2. 1 H NMR (300 MHz, CDCl3) 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]

[0059] 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl(sulfanyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-[3-(trifluoromethylthio)phenyl]piperazine (1.57 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0060] White solid. Yield: 64% (1.83 g); mp 97.8-99.2 °C; TLC: R f = 0.48 (DCM:MeOH (9:0.5; v / v)); C 23 H 22 F3N3O3S (477.50), monoisotopic mass: 478.13. UPLC (99% purity): t R = 7.55 min, (M+H) + 478.1. 1 H NMR (500 MHz, CDCl3) 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). 13 C NMR (126 MHz, CDCl3) 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]

[0061] 1-(2-(4-([1,1'-biphenyl]-3-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-(biphen-3-yl)piperazine (1.43 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0062] White solid. Yield: 82% (2.23 g); mp 114.1-115.4 °C; TLC: R f = 0.4 (DCM:MeOH (9:0.5; v / v)); C 28 H 27 N3O3 (453.54), monoisotopic mass: 453.20. UPLC (100% purity): t R = 7.43 min, (M+H) + 454.2. 1 H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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]

[0063] 1-(1-(4-fluorophenyl)-2-oxo-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-(4-fluorophenyl)acetic acid (1.51 g, 6 mmol, 1 equivalent) and 1-[3-(trifluoromethoxy)phenyl]piperazine (1.38 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0064] White solid. Yield: 73% (2.03 g); mp 88.8-90.7 °C; TLC: R f = 0.63 (DCM:MeOH (9:0.5; v / v)); C 23 H 21 F4N3O3 (463.43), monoisotopic mass: 463.15. UPLC (100% purity): t R = 7.05 min, (M+H) + 464.2. 1 H NMR (300 MHz, CDCl3) 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). 13 C NMR (75 MHz, CDCl3) 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]

[0065] 1-(2-(4-(naphth-2-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-(naphth-2-yl)piperazine (1.27 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0066] White solid. Yield: 79% (2.02 g); mp 197.1-198.5 °C; TLC: R f = 0.71 ((DCM:MeOH (9:0.5; v / v)); C 26 H 25 N3O3 (427.50), monoisotopic mass: 427.19. UPLC (100% purity): t R =6.97 minutes (M+H) + 428.2. 1 H NMR (300 MHz, CDCl3) 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]

[0067] 1-(2-(4-(benzo[b]thiophen-5-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-(benzo[b]thiophen-5-yl)piperazine (1.30 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0068] White solid. Yield: 79% (2.05 g); mp 164.1-165.3 °C; TLC: R f = 0.66 ((DCM:MeOH (9:0.5; v / v)); C 24 H 23 N3O3S (433.53), monoisotopic mass: 433.15. UPLC (100% purity): t R = 6.62 minutes, (M+H) + 434.1. 1 H NMR (300 MHz, CDCl3) 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]

[0069] 1-(2-(4-(1,2-benzoxazol-5-yl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 5-(piperazin-1-yl)benzo[d]isoxazole (1.22 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0070] White solid. Yield: 57% (1.43 g); mp 186.4-187.8 °C; TLC: R f = 0.58 (DCM:MeOH (9:0.5; v / v)); C 23 H 22 N4O4 (418.45), monoisotopic mass: 418.16. UPLC (98% purity): t R = 7.25 min, (M+H) + 419.1. 1 H NMR (300 MHz, CDCl3) 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]

[0071] 1-(2-(4-(3-chlorophenyl)piperidin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 4-(3-chlorophenyl)piperidine (1.17 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0072] White solid. Yield: 74% (1.83 g); mp 111.8-113.4 °C; TLC: R f = 0.43 (DCM:MeOH (9:0.5; v / v)); C 23 H 23 ClN2O3 (410.90), monoisotopic mass: 410.14. UPLC (100% purity): t R = 7.05 min, (M+H) + 411.1. 1 H NMR (300 MHz, CDCl3) 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]

[0073] 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperidin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 1-[3-(trifluoromethyl)phenyl]piperidine (1.37 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0074] White solid. Yield: 85% (2.26 g); mp 100.1-101.5°C; TLC: R f = 0.45 (DCM:MeOH (9:0.5; v / v)); C 24 H 23 F3N2O3 (444.45), monoisotopic mass: 444.17. UPLC (100% purity): t R = 7.26 minutes, (M+H) + 445.1. 1H NMR (300 MHz, CDCl3) 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]

[0075] 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperidin-1-yl)ethyl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-2-phenylacetic acid (1.40 g, 6 mmol, 1 equivalent) and 4-[3-(trifluoromethoxy)phenyl]piperidine (1.45 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0076] White solid. Yield: 79% (2.18 g); mp 112.1-113.2 °C; TLC: R f = 0.47 (DCM:MeOH (9:0.5; v / v)); C 24 H 23 F3N2O4 (460.45), monoisotopic mass: 460.16. UPLC (100% purity): t R = 7.12 min, (M+H) + 461.1. 1 H NMR (300 MHz, CDCl3) 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]

[0077] 1-(1-oxo-3-phenyl-1-(4-phenylpiperazin-1-yl)prop-2-yl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 equivalent) and 1-phenylpiperazine (0.97 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0078] White solid. Yield: 87% (1.13 g); mp 121.7-123.2 °C; TLC: R f = 0.62 (DCM:MeOH (9:0.5; v / v)); C 23 H 25 N3O3 (391.47), monoisotopic mass: 392.19. UPLC (100% purity): t R = 6.22 minutes, (M+H) + 392.1. 1 H NMR (300 MHz, CDCl3) 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). 13 C NMR (75 MHz, CDCl3) 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]

[0079] 1-(1-(4-(3-chlorophenyl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 equivalent) and 1-(3-chlorophenyl)piperazine (1.40 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0080] White solid. Yield: 87% (1.23 g); mp 114.3-116.2 °C; TLC: R f = 0.80 (DCM:MeOH (9:0.5; v / v)); C 23 H 24 ClN3O3 (425.91), monoisotopic mass: 426.15. UPLC (100% purity): R = 6.97 min, (M+H) + 426.1. 1 H NMR (300 MHz, CDCl3) 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). 13 C NMR (75 MHz, CDCl3) 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]

[0081] 1-(1-oxo-3-phenyl-1-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)propan-2-yl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 equivalent) and 1-[3-(trifluoromethyl)phenyl]piperazine (1.38 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0082] White solid. Yield: 84% (1.59 g); mp 126.1-127.2 °C; TLC: R f = 0.72 (DCM:MeOH (9:0.5; v / v)); C 24 H 24 F3N3O3 (459.47), monoisotopic mass: 460.18. UPLC (100% purity): R = 7.22 min, (M+H) + 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]

[0083] 1-(1-(4-([1,1'-biphenyl]-3-yl)piperazin-1-yl)-1-oxo-3-phenylpropan-2-yl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 equivalent) and 1-(biphenyl-3)piperazine (1.43 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0084] White solid. Yield: 88% (1.46 g); mp 119.1-120.0 °C; TLC: R f= 0.77 (DCM:MeOH (9:0.5; v / v)); C 29 H 29 N3O3 (467.57), monoisotopic mass: 467.22. UPLC (100% purity): R = 7.63 min, (M+H) + 468.2. 1 H NMR (300 MHz, CDCl3) 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). 13 C NMR (75 MHz, CDCl3) 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]

[0085] 1-(1-oxo-3-phenyl-1-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)propan-2-yl)pyrrolidine-2,5-dione The compound was prepared according to the procedure described in Example 2. 2-(2,5-dioxopyrrolidin-1-yl)-3-phenylpropanoic acid (1.48 g, 6 mmol, 1 equivalent) and 1-[3-(trifluoromethoxy)phenyl]piperazine (1.48 g, 6 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0086] White solid. Yield: 83% (1.32 g); mp 104.4-105.5 °C; TLC: R f= 0.71 (DCM:MeOH (9:0.5; v / v)); C 24 H 24 F3N3O4 (475.47), monoisotopic mass: 475.17. UPLC (100% purity): t R = 7.40 min, (M+H) + 476.1. 1 H NMR (300 MHz, CDCl3) 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]

[0087] Determination of anticonvulsant activity in vivo in mice Male Swiss albino mice (CD-1) weighing 18-26 g were used. All procedures were performed in accordance with applicable Polish and international guidelines on the ethics of animal testing after receiving appropriate institutional approval. Substances were administered intraperitoneally (ip) in a 1% aqueous solution of Tween as a single injection in a volume of 10 ml / kg 30 min before a given test. Screening was performed in groups of 4 mice. The mean effective dose (ED) for a given test was 1.0 mg / kg. 50 ), and the toxic dose in the rotarod test (TD 50 ) were estimated based on results obtained in 3–4 groups of 6 mice. All studies were performed according to procedures described in the expert literature. [Example]

[0088] Maximal electroshock seizure test (MES) In the MES test, seizures are induced by the continuous electric stimulation of 0.2 seconds duration, 500V voltage, and 25mA intensity.The electric stimulation is generated by using an electric shock generator (Rodent shocker, Type221, Hugo Sachs Elektronik, Germany) and delivered to animals using electrodes placed on the ear pinna.The test is carried out 30 minutes after intraperitoneal administration of various doses of compound. During the experiment, the number of animals that experienced seizure episodes in the form of tonic extension of the hind limbs was counted (Kaminski, K.; Rapacz, A.; Luszczki, JJ; Latacz, G.; Obniska, J.; Kiec-Kononowicz, K.; Filipek, B., Bioorg. Med. Chem. 2015, 23, 2548-2561; Castel-Branco, MM; Alves, GL; Figueiredo, IV; Falcao, AC; Caramona, MM, Methods Find. Exp. Clin. Pharmacol. 2009, 31, 101-106). [Example]

[0089] Psychomotor seizure test (6 Hz test) In the 6 Hz test, seizures were induced by electrical stimulation at 32 mA and / or 44 mA and a frequency of 6 pulses per second. Electrical pulses were generated using an electric shock generator (ECT Unit 57800; Ugo Basile, Gemonio, Italy) and delivered to the animals using ocular electrodes. Before the test began, the surface of the eye was gently moistened with a local anesthetic solution (1% lidocaine solution). The test was performed 30 minutes after intraperitoneal administration of various doses of compounds. Electrical pulses were delivered consecutively for 3 seconds, and the animals were observed for 10 seconds. During this period, observations were made for rearing-related immobility or stunnedness, forelimb clonus, vibrissae twitching, and Straub's tail. These symptoms persisted throughout the observation period, indicating the occurrence of psychomotor seizures in the mice. Mice that resumed normal behavior within 10 seconds of stimulation were considered protected (Barton, ME; Klein, BD; Wolf, HH; White, HS, Epilepsy Res. 2001, 47, 217-227; Wojda, E.; Wlaz, A.; Patsalos, PN; Luszczki, JJ, Epilepsy Res. 2009, 86, 163-174). [Example]

[0090] Subcutaneous pentylenetetrazol seizure test (scPTZ) In the scPTZ test, seizures were induced by subcutaneous administration of pentylenetetrazole (PTZ) at a dose of 100 mg / kg. This caused clonic seizures accompanied by loss of the righting reflex. The test compound was administered 30 minutes before the experiment. After PTZ administration, animals were individually placed in transparent containers and observed for 30 minutes for the onset of clonic seizures. During this period, the latency to the first onset of clonic seizures, defined as a generalized clonus lasting at least 3 seconds accompanied by loss of the righting reflex, and the number of seizure episodes during the test period were observed and compared with the control group. The absence of clonic convulsions within the observed period was interpreted as the ability of the compound 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; Laczkowski, K.; Salat, K.; Misiura, K.; Podkowa, A.; Malikowska, N., J. Enzyme Inhib. Med. Chem. 2016, 31, 1576-82). [Example]

[0091] Effects on motor coordination in mice in the rotarod test The influence of the tested compounds on motor coordination was evaluated by rotarod test (apparatus used: May Commat, RR 0711 Rota Rod, Turkey). Mice were trained the day before the actual experiment. They were individually placed on a 2 cm diameter rod that rotated at 10 revolutions per minute (rpm). During each training session, animals remained on the rod for 3 minutes. The experiment was carried out 30 minutes after administration of the compound. Motor coordination was tested for 60 seconds, with a rotating rod speed of 10 rpm. Motor impairment was defined as being unable to remain on the rotating rod for 1 minute. The average time spent on the rod was calculated for each experimental group (Dunham, NW; Miya, TA; Edwards, LD, J. Am. Pharm. Assoc. 1957, 46, 64-66; Laczkowski, K.; Salat, K.; Misiura, K.; Podkowa, A.; Malikowska, N., J. Enzyme Inhib. Med. Chem. 2016, 31, 1576-82). [Example]

[0092] statistical analysis ED along with the corresponding 95% confidence intervals 50 (effective dose) and TD 50 The (toxic dose) values were calculated based on the Litchfield and Wilcoxon method (Litchfield, JT, Wilcoxon, F., 1949, A simplified method of evaluating dose-effect experiments. J. Pharmacol. Exp. Ther. 96, 99-113). One-way ANOVA analysis of variance and Dunnett's post hoc test (multiple comparison test) were used to perform statistical evaluation of the results. A significant level of p<0.05 was considered statistically significant. [Example]

[0093] Anticonvulsant activity test results The compounds of the present invention demonstrated 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 the 30 min time point, they protected 50-100% of the animals tested. The most potent protection was observed for compounds containing an electron-withdrawing substituent, preferably Cl, CF3, OCF3, SCF3, CHF2, or a phenyl substituent, at position 3 of the aromatic ring connected to the piperazine moiety, where k is preferably 0. Table 1 shows the pharmacological screening data for selected substances.

[0094] [Table 1]

[0095] The above tests were carried out on racemic mixtures of compounds according to the invention.

[0096] 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 mice in the MES test, the 6 Hz (32 mA and 44 mA) test, and the scPTZ test during the screening test (0.5 hour time point).

[0097] [Table 2]

[0098] The results obtained confirmed that the compounds of the present invention, especially compound 6, have a strong protective effect and a significantly better protection index compared to the model AED-valproic acid. In particular, valproic acid is known to have a wide spectrum of therapeutic applications. [Example]

[0099] Determination of antinociceptive activity in vivo in mice Tests were performed using male albino Swiss mice (CD-1) weighing 18-25 g. All procedures were performed in accordance with Polish and international guidelines on the ethics of animal testing after receiving appropriate institutional approval. Test groups consisted of 8-10 animals. Test and reference substances were administered intraperitoneally as suspensions in a 1% aqueous solution of Tween 30 min before a given test. All tests / models were reviewed in expert literature: formalin test (Beirith, A.; Santos, AR; Calixto, JB; Rodrigues, AL; Creczynski-Pasa, TB, Eur. J. Pharmacol. 1998, 345, 233-245), 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), model of oxaliplatin-induced neuropathic pain - von The Frey test was performed according to the procedure described in (Salat, K.; Cios, A.; Wyska, E.; Salat, R.; Mogilski, S.; Filipek, B.; Wieckowski, K.; Malawska, B., Pharmacol. Biochem. Behav. 2014, 122, 173-181). [Example]

[0100] Determination of analgesic activity in the formalin test Pain was induced by subplantar injection of 20 μL of 2.5% formalin solution into the right hind paw of the mice. The animals were placed in separate transparent observation chambers for 30 minutes. The measurement was the sum of the time spent licking and biting the paw injected with formalin solution. The nociceptive response time was calculated during the first 5 minutes after formalin injection (first phase of the test: acute pain) and at time intervals of 15-20 minutes, 20-25 minutes, and 25-30 minutes after administration (second phase of the test: inflammatory pain). The observed inhibition of the nociceptive response - a reduction in the time spent licking and biting the paw - was interpreted as an analgesic effect of the tested compound. Based on the results obtained, the ED 50 The dose (the dose that reduces nociceptive reaction time by 50%) was calculated.The reference compound in this test is valproic acid, which is intraperitoneally administered at the dose of 100mg / kg, 150mg / kg and 200mg / kg.Compound 6 is administered at the dose of 10mg / kg, 20mg / kg and 30mg / kg.

[0101] Compound 6 showed differential 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 phases of the test, respectively. Compound 6 reduced nociceptive response time in the first phase of the formalin test, which corresponds to acute pain, at all doses tested, with statistically significant effects observed at the two highest doses. The ED of Compound 6 in the first phase of the test was 50 The ED value was 28.50 mg / kg. In the second phase of the study, which corresponds to persistent inflammatory pain, Compound 6 statistically significantly reduced the duration of the nociceptive response at all doses used. The ED value in the second phase of the study for this compound was 50 The value was 12.40 mg / kg (Figure 3).

[0102] Valproic acid (VPA) did not exhibit analgesic activity in the first phase of the study at any of the doses tested. In the second phase of the VPA study, nociceptive response times were reduced at all doses used, and the ED 50 The value was 132.90 mg / kg (Figure 3). [Example]

[0103] Determination of analgesic activity in the capsaicin pain model This test evaluates the time that mice lick and / or bite their hind paws after subplantar injection of 1.6 μg of capsaicin dissolved in 20 μl of a mixture containing 0.9% saline and ethanol (5% of the final volume). Observations were carried out for 5 minutes after capsaicin administration. Test compounds were administered intraperitoneally 30 minutes before capsaicin administration. Inhibition of nociceptive responses—reduction in paw licking or biting time was a measure of the antinociceptive activity of the tested compounds.

[0104] Valproic acid (VPA) was the reference compound in this study. 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. The 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.

[0105] Compound 6 statistically significantly reduced nociceptive response time at 20 mg / kg and 30 mg / kg, and ED 50 was 17.9 mg / kg (Figure 4).

[0106] 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 administration of 200 mg / kg (FIG. 4). [Example]

[0107] Model of oxaliplatin-induced neuropathic pain - Determination of analgesic activity in the von Frey assay Oxaliplatin (OXPT) was dissolved in a 5% glucose solution and then administered intraperitoneally to mice. A single dose of 10 mg / kg was used. Tactile and thermal (cold sensation) allodynia associated with oxaliplatin-induced neuropathy is characterized by two phases: the early phase is acute and develops shortly after OXPT administration, while the late (chronic) phase (associated with neuronal damage) symptoms develop several days later. Behavioral testing of mice with OXPT-induced neuropathy was performed 7 days after administration, i.e., during the late phase of neuropathy.

[0108] The effects of the tested compounds on tactile allodynia were determined using the von Frey test. Animals were individually placed in mesh-bottom cages 60 minutes before the start of the experiment to allow adaptation to the novel environment. The pain threshold to mechanical stimulation was assessed using an electronic von Frey apparatus (Electronic Von Frey, Bioseb, France). A von Frey fiber was applied to the underside of the right paw of the mouse with increasing pressure. Upon crossing the pain threshold, the paw was withdrawn, and the mechanical pressure that elicited a nocifensive response was subsequently recorded. Measurements were performed three times for each mouse, with at least 30 seconds between measurements, and the results were averaged. The entire test was performed three times: before OXPT administration to determine the baseline pain threshold; before administration of the test compound 7 days after OXPT administration to evaluate the onset of neuropathy by establishing a new pain threshold; and 30 minutes after compound administration to determine the effect on the onset of neuropathy.

[0109] The effect of test compounds on thermal allodynia was assessed in the cold plate test using a specialized apparatus: Cold / Hot Plate, Bioseb, France. Animals were individually placed on a metal plate cooled to 2°C using a thermostatic device. The observed nociceptive responses of the animals included licking and / or characteristic hind paw lifting or jumping. The observation time was set to 60 seconds to eliminate the potential risk of tissue damage and minimize animal discomfort. As with the Von Frey test, measurements were performed in triplicate.

[0110] Compound 6 and valproic acid as a reference AED were administered intraperitoneally as suspensions 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.

[0111] Injection of OXPT in mice caused the development of neuropathy, resulting in a significant and statistically significant reduction in pain threshold, as measured by the von Frey method. Pain sensitivity thresholds decreased in OXPT-treated mice from 3.18 ± 0.06 to 3.36 ± 0.10 g in healthy mice to levels ranging from 1.89 ± 0.04 to 1.94 ± 0.14 g. The results demonstrate a statistically significant analgesic effect of the tested compound 6. While the mean pain sensitivity threshold in the control group was 3.36 ± 0.10 g, after OXPT administration 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 elevated the pain threshold to 2.87 ± 0.12 g (85.41% of the initial value), indicating an inhibitory effect on the development of mechanical allodynia, even at low doses. A dose of 20 mg / kg of compound 6 increased the pain sensitivity threshold to 3.83 ± 0.13 g, which is 113.98% of the initial value. A dose of 30 mg / kg increased the pain threshold to 4.17 ± 0.17 g, which is 124.10% of the initial value. The results show that compound 6 is highly effective in suppressing the onset of mechanical allodynia, which is a consequence of neuronal damage caused by the chemotherapeutic agent OXPT (Figure 5A).

[0112] The mean pain sensitivity threshold in the control group for a reference compound (valproic acid, VPA) was 2.62 ± 0.06 g, which decreased to 1.78 ± 0.04 g after administration of OXPT. Administration of VPA at a dose of 150 mg / kg caused a maximum increase in the pain threshold of 3.97 ± 0.30 g, while doses of 100 mg / kg and 50 mg / kg body weight allowed achieving an increase in the mean pain threshold of 3.18 ± 0.14 g and 2.75 ± 0.06 g, respectively (Figure 5B).

[0113] Compound 6 also significantly enhanced thermal allodynia sensitivity in the cold plate test (FIG. 5C). [Example]

[0114] In vitro affinity and functionality testing In vitro affinity and functionality tests performed on the most active substance 6, representing a compound according to formula (II), showed that its mechanism of action is related to its effect on nerve conduction through interaction with voltage-gated sodium channels (site 2) and calcium channels (dihydropyridine, diltiazem, and verapamil binding sites). A unique feature of compound 6, representing a compound according to formula (II) of the present invention, is its inhibition of calcium currents by blocking transient receptor potential vanilloid type 1 (TRPV1). This effect has not been described for known AEDs. TRPV1 receptor antagonism can determine the antinociceptive effect of the compounds disclosed herein. The role of TRPV1 receptors in the transmission of pain stimuli is well documented in the expert literature (Szallasi, A.; Cortright, DN; Blum, CA; Eid, SR, Nat. Rev. Drug. Discov. 2007, 6, 357-372). The compounds according to the present invention are characterized by a complex mechanism of action, which has not been described for known anticonvulsants. However, it should be emphasized that further in vitro studies may reveal additional molecular targets involved in the pharmacological action of the presently claimed substances. The results of binding studies (sodium channel, calcium channel) and functional studies (TRPV1 receptor) for compound 6 are shown in Table 3.

[0115] [Table 3A] [Example]

[0116] In vitro electrophysiology studies Experiments were performed in accordance with institutional and international guidelines for 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 solution. The sectioning and preincubation methodology has been previously described (Szulczyk, B.; Nurowska, E., Biochem. Biophys. Res. Commun. 2017, 491, 291-295). Slices containing the prefrontal cortex were enzymatically and mechanically dispersed. Single prefrontal cortical pyramidal neurons were visualized using an inverted microscope (Nikon). Sodium currents were evoked by rectangular depolarizing stimuli. The potential between depolarizing stimuli was maintained at -65 mV.

[0117] The intracellular solution in the pipette contained (in mM): CsF (110), NaCl (7), EGTA (3), HEPES-Cl (10), MgCl2 (2), Na2ATP (4) (pH 7.4 and osmolarity 290 mOsm).

[0118] The extracellular solution used to wash neurons contained (in mM): NaCl (30), choline chloride (90), TEA-Cl (30), CaCl2 (2), MgCl2 (2), glucose (15), HEPES (10), LaCl3 (0.001), and CdCl2 (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, the pipette capacitance was compensated by the amplifier.

[0119] After the patch membrane was disrupted by suction or electrical stimulation, the membrane capacitance was compensated. The access resistance was between 5 and 7 MΩ. 80% series resistance compensation was used. Leakage currents were subtracted from the recorded currents. Recordings were performed at room temperature. Voltage-gated potassium currents were blocked by TEA-Cl in the extracellular solution. Voltage-gated calcium currents were blocked by cadmium and lanthanum ions in the extracellular solution. The membrane potential of the neuron was maintained at -65 mV. Substance 6 was administered extracellularly (to the whole bath).

[0120] The results confirmed the inhibitory effect of Compound 6 by rapidly activating and rapidly inactivating voltage-gated sodium channels in prefrontal cortical pyramidal neurons (tests were performed at a concentration of 100 μM). Maximal currents were elicited by rectangular depolarizing stimuli lasting 20 ms. The potential between depolarizing stimuli was maintained at -65 mV. Control recordings were performed for 2 min, test substances were administered for 3 min, and washout currents were recorded for 5 min. Recorded currents were normalized to the control current values. Substance 6 blocked the maximum amplitude of sodium currents by a maximum of 0.59 ± 0.08 compared to the control (1.0, p < 0.001). After washout, the current amplitude partially recovered to the control value (0.79 ± 0.07, n = 5). An example of sodium current recordings and averaged results is shown in Figure 6. [Example]

[0121] Evaluation of ADMETox parameters in in vitro studies The ADME-Tox parameters of compound 6 were evaluated by in vitro methods using recombinant enzymes, human and mouse liver microsomes, and eukaryotic cell lines.

[0122] Metabolic stability. The metabolic stability of compound 6 was evaluated using human liver microsomes (HLM). The internal clearance value CL intwas calculated by monitoring the change in compound concentration in the presence of microsomes per time unit according to the procedure proposed by Obach RS (Obach, RS, Drug Metab. Dispos. 1999, 27, 1350-1359). Based on the data obtained, a very low clearance value of compound 6 after incubation with HLM was found, and CL int The HCl concentration reached 5.8 ml / min / kg, demonstrating the predicted high stability in the human body. UPLC analysis of the metabolism of compound 6 after incubation with HML revealed that it was metabolized to three metabolites, M1–M3 (Figure 7). Based on the UPLC / MS data, metabolite M1 was formed by dehydrogenation of the piperazine ring, M2 by hydroxylation of the phenyl substituent linked to the piperazine, while M3 was formed with high certainty as a result of hydroxylation of the lateral phenyl group concomitant with reduction of the ketone group in the imide fragment to a hydroxyl group (Figure 7).

[0123] Metabolic Stability Test—Methodology. Metabolic stability testing for compound 6 was performed using HLM (Promega, Madison, WI, USA). For this purpose, 10 μL of compound 6 at a concentration of 1000 μM was diluted with 132 μL of Tris-HCl buffer (100 mM, pH 7.4), followed by the addition of 8 μL of the appropriate microsomes. The reaction mixture was preincubated at 37°C for 5 min, followed by the addition of 50 μL of NADPH Regeneration System provided by Promega (Madison, WI, USA). After mixing, the entire mixture was incubated at 37°C for 120 min. To complete the reaction, 200 μL of cold methanol was added to the tube and centrifuged. The supernatant was subjected to UPLC / MS analysis, including fragmentation analysis. Four mixtures of 6 and HLM were prepared to measure the internal clearance CL. intwas determined. Each of these reactions was completed at different time points, 5, 15, 30, and 45 minutes, by adding cold methanol containing 50 μM of internal standard. Then, following literature guidelines (Obach, RS, Drug Metab. Dispos. 1999, 27, 1350-1359), a regression equation was determined based on a plot of the relationship between the peak height from 6 and the height of the internal standard, and the reaction rate constant k was calculated. The constant k was then substituted into equation (1).

[0124]

number

[0125] Then, the calculated t 1 / 2 The values were substituted into equation (2).

[0126]

number

[0127] Effect on Pgp Activity. P-glycoprotein (Pgp) is an integral plasma membrane protein that acts as an ATP-dependent burst pump to actively remove xenobiotics and can cause drug-drug interactions. Pgp plays an important role in drug absorption in the gastrointestinal tract and across the blood-brain barrier. The effect of compound 6 on Pgp activity was tested using a commercial bioluminescent Pgp-Glo™ Assay System (Promega, Madison, WI, USA). The test procedure is based on measuring changes in the level of ATP consumed by membranes containing recombinant Pgp protein in the presence of the test compound. Results are presented as % of baseline activity and compared with reference compounds: the selective Pgp inhibitor Na3VO4 and the stimulant verapamil. Compound 6 exhibited 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 was observed at 50 μM (Figure 8).

[0128] Effect on Pgp Activity—Methodology. The test was performed according to the bioluminescent Pgp-Glo™ Assay System test protocol provided by Promega (Madison, WI, USA). The enzymatic reaction was performed in a Nunc™ MicroWell™ 96-well white plate from Thermo Scientific (Waltham, MA, USA). Bioluminescence was measured using a PerkinElmer multispecific EnSpire plate reader (Waltham, MA, USA). After using the Na3VO4 Pgp inhibitor (inducing 100% inhibition), the signal increased relative to the control sample, indicating inhibition of ATP consumption by Pgp, the so-called base activity. The calculated difference in luminescence values between the inhibitor-treated sample and the control sample was taken as 100% Pgp base activity and served as the negative control in the test. The reference compounds Na3VO4 and verapamil were used at 100 μM and 200 μM, respectively, according to the manufacturer's instructions. Compound 6 was tested at concentrations of 50 and 100 μM, obtained after dilution of a concentrated stock solution (10 mM) in DMSO in reaction buffer. Incubation of the compound with Pgp-containing membranes was carried out at 37°C for 40 minutes, followed by bioluminescence measurements to determine the extent of ATP consumption by Pgp. Statistical significance was calculated by one-way ANOVA analysis of variance and the Bonferroni method using GraphPad Prism 5. Compounds were tested in triplicate.

[0129] Effect of compound 6 on cytochrome P-450 3A4 and 2D6 activity. The investigation was carried out as described in the literature (Socala, K.; Mogilski, S.; Pierog, M.; Nieoczym, D.; Abram, M.; Szulczyk, B.; Lubelska, A.; Latacz, G.; Doboszewska, U.; Wlaz, P.; Kaminski, K., ACS Chem. Neurosci. 2018, doi:10.1021 / acschemneuro.8b00476; Latacz, G.; Lubelska, A.; Jastrzebska-Wiesek, M.; Partyka, A.; Sobalo, A.; Olejarz, A.; Kucwaj-Brysz, K.; Satala, G.; Bojarski, AJ; Wesolowska, A.; Kiec-Kononowicz, K.; Handzlik, J., Chem. Biol. Drug. Des. 2017, 90, 1295-1306), using the commercially available luminescent assays CYP3A4 P450-Glo™ and CYP2D6 P450-Glo™ from Promega (Madison, WI, USA). The CYP isoforms selected for testing are involved in the metabolism of approximately 40-50% of marketed drugs, and their stimulation or inhibition determines the majority of metabolic drug-drug interactions. The results show that at a concentration of 10 μM, compound 6 has no effect on CYP3A4 activity (Figure 9A) and a very weak stimulatory effect on CYP2D6 (Figure 9B). Overall, the results indicate a low likelihood of potential metabolic interactions caused by 6.

[0130] In vitro hepatotoxicity evaluation. Tests were performed using the hepatocellular carcinoma HepG2 liver cancer cell line, which was used to evaluate the hepatotoxicity of the compounds in vitro. The classic MTS colorimetric assay from Promega (Madison, WI, USA) was used to investigate the effect of 6 on HepG2 cell viability and proliferation. The compound was tested at four concentrations ranging from 0.1 to 100 μM. Doxorubicin at a concentration of 1 μM was used as the reference cytostatic agent. In addition, the reference mitochondrial toxin carbonyl cyanide m-chlorophenylhydrazone (CCCP) at a concentration of 10 μM was also used (Figure 10). Hepatotoxicity testing after 72 h of incubation of HepG2 cells with compound 6 showed a statistically significant (p<0.05) reduction in cell viability only at the maximum concentration used in the test, 100 μM (Figure 10). In addition, cell viability decreased to just 84% of the control, indicating a slight toxic effect of this compound on the HepG2 cell line. To specifically expose hepatocytes to the potential toxic effects of xenobiotics, an additional test was performed on the HepG2 cell line in the form of a luminescent measurement of cellular ATP levels after a brief 3-hour exposure to compound 6 at concentrations ranging from 1 to 100 μM. For this purpose, a commercial CellTiter-Glo Luminescent Cell Viability Assay from Promega (Madison, WI, USA) was used. The purpose of the test was to examine the effect of the compound on mitochondrial respiration in hepatocellular carcinoma cells. The reference point was CCCP, the 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 maximum concentration used, 100 μM. This indicates a very low risk of hepatotoxic effects of compound 6 (Figure 11).

[0131] In vitro hepatotoxicity assessment - methodology. The HepG2 hepatocellular carcinoma cell line (ATCC HB-8065) was used in the study. The HepG2 line was incubated in "Modified Eagle's Medium" (MEM) culture medium supplemented with 2 mM glutamine and 10% FBS from Gibco (Carlsbad, CA, USA). The cells were incubated at 37°C in an atmosphere containing 5% CO2. Cell viability was tested using the CellTiter96® AQueous Non-Radioactive Cell Proliferation Assay (MTS) supplied by Promega (Madison, WI, USA). Prior to the study, cells were plated at 1.5 x 10 per well. 4 Cells were placed in Thermo Scientific Nunc™ 96-well clear culture plates (Waltham, MA, USA) at a concentration of 1 / 3 of a cell and incubated for 24 hours. A 10 mM stock solution of compound 6 was then diluted in the appropriate culture medium and added to the cells at final concentrations ranging from 0.1 to 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 hours of incubation at 37°C in an atmosphere containing 5% CO2, the compound-containing medium was removed, followed by the addition of fresh medium containing the diluted MTS reagent. The plates were further incubated for 2–3 hours, after which absorbance measurements were taken at 490 nm using an EnSpire PerkinElmer (Waltham, MA, USA) reader. Statistical significance was calculated using one-way ANOVA analysis of variance and the Bonferroni method. Compounds were tested in quadruplicate.

[0132] ATP levels in HepG2 cells were tested using the CellTiter-Glo Luminescent Cell Viability Assay from Promega (Madison, WI, USA). Prior to testing, cells were plated in white 96-well clear-bottom culture plates from Corning (Tewksbury, MA, USA) at 1.5 × 10 cells per well. 4The cells were then incubated at 37°C in an atmosphere containing 5% CO2. Compound 6 was added to the plate at three final concentrations of 1, 10, and 100 μM, CCCP at 10 μM, and DX at 1 μM in a volume of 100 μL. The plate was incubated at 37°C in 5% CO2 for 3 hours. After adding 100 μl of CellTiter-Glo Luminescent Cell Viability Assay to the culture, luminescence measurements were performed using an EnSpire PerkinElmer (Waltham, MA, USA) reader. Statistical significance was calculated by one-way ANOVA and Bonferroni analysis using GraphPad Prism5. All substances were tested in quadruplicate. [Example]

[0133] Preparation of Selected Enantiomers of Compounds According to the Invention The enantiomers of the compound according to formula (II) of the present invention can be obtained by applying a four-step procedure using commercially available tert-butoxycarbonyl (Boc) D- or L-amino acid derivatives (R or S absolute configuration, respectively) as starting materials. Enantiomers are obtained for selected compounds as described by formula (II), where k=0 and A and B have the same meaning as in the case of the racemic mixture of formula (II).

[0134] A general scheme for the synthesis of enantiomers of compounds according to formula (II) is shown in FIG.

[0135] In the first step, a given piperazine derivative is condensed with the corresponding Boc-D- or Boc-L-amino acid derivative to give an intermediate of formula (VII), followed by a deprotection reaction to form the amine derivative (VI). In the next step, compound (VI) is condensed with succinic anhydride to give an intermediate with an amide-acid structure (V), which is then cyclized to form compound R-(II) or S-(II). The asymmetric synthesis proceeds while retaining the absolute configuration confirmed by crystallography.

[0136] Examples of syntheses and physicochemical and spectral data for selected intermediates (VII, VI, and V according to Figure 12) are described below. [Example]

[0137] tert-Butyl-(R)-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)carbamate (VII) Boc-D-phenylglycine (1.25 g, 5 mmol, 1 equiv.) was dissolved in 20 mL of DCM, followed by the addition of DCC (1.55 g, 7.5 mmol, 1.5 equiv.). After 30 min, 1-(3-(trifluoromethyl)phenyl)piperazine (1.15 g, 5 mmol, 1 equiv.) dissolved in 5 mL of DCM was added. The reaction was allowed to stir at room temperature for 4 h. After this time, the DCM was evaporated to dryness. Intermediate VII was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0138] Light oil. Yield: 78% (1.81g); TLC:R f = 0.62 (DCM:MeOH (9:0.5; v / v)); C 24 H 28 F3N3O3 (463.50), monoisotopic mass: 463.21. UPLC (100% purity): t R =8.40 min. (M+H) + 464.2. [Example]

[0139] (R)-2-amino-2-phenyl-1-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethan-1-one (VI) 5 mL of TFA was added to a solution of tert-butyl-(R)-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)carbamate (VII, 1.39 g, 3 mmol, 1 equiv.) in DCM (50 mL) and stirred for 2 h. The reaction mixture was then neutralized with 25% NH4OH solution and extracted with DCM (3 x 50 mL). The organic layer was dried over anhydrous Na2SO4 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.

[0140] Yellow oil. Yield: 95% (1.03g);C 19 H 20 F3N3O (363.38), monoisotopic mass: 363.16. UPLC (purity >99.9%): R =4.96 minutes. (M+H) + 364.1. [Example]

[0141] (R)-4-oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)amino)butanoic acid (V) Succinic anhydride (0.28 g, 2.8 mmol, 1 equiv.) 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 equiv.) in AcOEt (50 mL) and the mixture was stirred for 30 min. After this time, the solvent was evaporated to dryness. The compound was obtained in solid form after washing with EtO.

[0142] White solid. Yield: 87% (1.13 g); 23 H 24 F3N3O4 (463.46), monoisotopic mass: 463.17. UPLC (purity >99.9%): R =6.40 min. (M+H) + 464.2. [Example]

[0143] (R)-1-(2-(4-(3-chlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione ((R)-3) ZnCl (0.27 g, 2.0 mmol, 1 equiv.) was added to a 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 equiv.) in dry benzene (50 mL). The entire mixture was heated to 80 °C, and then a solution of HMDS (0.48 g, 0.62 mL, 3.0 mmol, 1.5 equiv.) in dry benzene (5 mL) was added dropwise over 30 min. The reaction was continued to stir under reflux for about 24 h and then concentrated under reduced pressure. After evaporation of 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 NaSO and then evaporated to dryness. The crude product was purified by column chromatography using a DCM:MeOH (9:0.3; v / v) eluent. The compound was obtained as a solid after washing with EtO.

[0144] White solid. Yield: 82%(0.67g);mp167.3~168.1℃;TLC:Rf=0.41(DCM:MeOH(9:0.3;v / v));C 22 H 22 ClN3O3 (411.89), monoisotopic mass: 411.13. UPLC (purity >99.9%): R = 6.70 min, (M+H) + 412.4. 1H NMR (500 MHz, CDCl3) 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). 13 C NMR (126 MHz, CDCl3) 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 R =40.25 minutes). [Example]

[0145] (R)-1-(2-(4-(3,5-dichlorophenyl)piperazin-1-yl)-2-oxo-1-phenylethyl)pyrrolidine-2,5-dione ((R)-4) The compound was prepared according to the 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 equivalent) was used as the starting material for the cyclization reaction. The crude product was purified by column chromatography using a DCM:MeOH (9:0.2; v / v) elution system.

[0146] White solid. Yield: 79% (0.70 g); mp 174.3-175.5 °C; TLC: R f = 0.43 (DCM:MeOH (9:0.2; v / v)); C 22 H 21Cl2N3O3 (446.33), monoisotopic mass: 445.10. UPLC (purity >99.9%): R = 7.59 min, (M+H) + 446.1. 1 H NMR (500 MHz, CDCl3) 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). 13 C NMR (126 MHz, CDCl3) 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 R =43.23 minutes). [Example]

[0147] (R)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione ((R)-6) The compound was prepared according to the 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 equivalent) was used as the starting material for the cyclization reaction. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0148] White solid. Yield: 80% (0.71 g); mp 189.1-190.5 °C; TLC: Rf = 0.35 (DCM:MeOH (9:0.5; v / v)); C 23 H 22 F3N3O3 (445.44), monoisotopic mass: 445.16. UPLC (purity >99.9%): R = 6.93 min, (M+H) + 446.2. 1 H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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 R =39.97 minutes). [Example]

[0149] (S)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione ((S)-6) The compound was prepared according to the 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 equivalent) was used as the substrate for the cyclization reaction. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0150] White solid. Yield: 78% (0.69 g); mp 188.9-190.5 °C; TLC: R f = 0.36 (DCM:MeOH (9:0.5; v / v)); C 23 H 22 F3N3O3 (445.44), monoisotopic mass: 445.16. UPLC (purity >99.9%): R = 6.94 min, (M+H) + 446.2. 1 H NMR (300 MHz, CDCl3) 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); 13 C NMR (75 MHz, CDCl3) 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 R =26.21 minutes). [Example]

[0151] (R)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethoxy)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione ((R)-10) The compound was prepared according to the 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 equivalent) was used as the starting material for the cyclization reaction. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0152] White solid. Yield: 77% (0.70 g); mp 168.2-169.1 °C; TLC: R f = 0.46 (DCM:MeOH (9:0.5; v / v)); C 23 H 22 F3N3O4 (461.44), monoisotopic mass: 461.16. UPLC (purity >99.9%): R = 7.18 min, (M+H) + 462.1. 1 H NMR (500 MHz, CDCl3) 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). 13 C NMR (126 MHz, CDCl3) 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 R =35.08 minutes). [Example]

[0153] (R)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl(sulfanyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione ((R)-12) The compound was prepared according to the 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 equivalent) was used as the starting material for the cyclization reaction. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system.

[0154] White solid. Yield: 86% (0.82 g); mp 155.1-155.8 °C; TLC: R f = 0.48 (DCM:MeOH (9:0.5; v / v)); C 23 H 22 F3N3O3S (477.50), monoisotopic mass: 477.13. UPLC (purity >99.9%): R = 7.54 min, (M+H) + 478.1. 1 H NMR (500 MHz, CDCl3) 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). 13C NMR (126 MHz, CDCl3) 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 R =34.82 minutes). [Example]

[0155] Special properties of enantiomers. The effect of the stereochemistry of the compounds according to the invention on their anticonvulsant activity was investigated. The anticonvulsant properties were evaluated according to the methods described above and the results are summarized in Tables 3 and 4.

[0156] [Table 3B]

[0157] [Table 4]

[0158] Based on the results obtained, it was unexpectedly found that the R enantiomer exhibits increased biological activity with a desirable profile compared to the S enantiomer.

[0159] In particular, for the R enantiomer, the following was found: - Less acute neurotoxicity in the rotarod test with respect to the racemate (TD in Tables 2 and 4, respectively) 50 (see - It was also unexpectedly found that the anticonvulsant effect was stereospecific. The enantiomer with the R configuration is characterized by stronger biological activity.

[0160] Metabolic Stability. The metabolic stability of (R)-6 was evaluated according to the methodology described above. Based on the data obtained, very low values of internal clearance of compound (R)-6 after incubation with HLM were found, and CL int = 2.4 mL / min / kg, indicating the predicted high stability in the human body. In addition, surprisingly, the determined clearance value was lower than that determined for the racemate, Compound 6 (CL int = 5.6), indicating a lower susceptibility of the enantiomers to metabolic changes. In addition, the results of UPLC analysis revealed that the (R)-6 enantiomer was preferably metabolized to two metabolites: the M1 metabolite formed by dehydrogenation of the piperazine ring and the M2 metabolite formed by hydroxylation of the phenyl substituent linked to the piperazine (Figure 13). In the case of the racemate, an additional M3 metabolite was observed, most likely obtained by hydroxylation of the lateral phenyl moiety and reduction of the keto group in the imide ring to a hydroxyl (Figure 7). [Example]

[0161] Preparation of Water-Soluble Salts of Compounds According to the Invention. The water-soluble salts of the compounds according to formula (I) of the present invention can be obtained by applying a six-step procedure using commercially available tert-butoxycarbonyl (Boc) amino acid derivatives as starting materials. The water-soluble salts are obtained for selected compounds described by formula (I), where k=0, D is a substituent selected from the group consisting of H, an amino group (-NH), an amino group substituted with one or two aliphatic substituents (especially -CH and / or -C H ), or an amino group that is part of a heterocycle, and A and B have the same meanings as in the compounds described by formula (II).

[0162] A general scheme for the synthesis of water-soluble salts of compounds according to formula (I) according to the present invention is shown in Figure 14. For the preparation of compounds of formula (I) where D is halogen, the procedure described for compounds of formula (II) according to Figure 2B is used, and the resulting compound is then converted into a water-soluble salt (preferably the hydrochloride salt) using methods described in the literature.

[0163] Steps i and ii are similar to the procedures described for the synthesis of the enantiomers. The amine derivative (VI) undergoes a condensation reaction with maleic anhydride to give a compound with an unsaturated amide-acid structure (VIII). Compound VIII is then cyclized to compound IX. In the next step, the compound of formula IX is subjected to an addition reaction with an appropriate primary or secondary amine. The desired compound according to formula (I) is then converted to a water-soluble salt (preferably the hydrochloride salt) using methods described in the literature.

[0164] Examples of syntheses and physicochemical and spectral data for selected intermediates (VIII, IX) and final products according to Figure 14 are described below. [Example]

[0165] 4-oxo-4-((2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)amino)but-2-enoic acid (VIII) Maleic anhydride (0.98 g, 10.0 mmol, 1 equiv.) 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 equiv.) in AcOEt (50 mL) and stirred for 30 min. After this time, the solvent was evaporated to dryness. The compound was obtained as a solid after washing with EtO.

[0166] White solid. Yield: 85% (3.76 g); 23 H 22 F3N3O4 (461.44), monoisotopic mass: 461.16. UPLC (purity = 96%): R=6.94 minutes. (M+H) + 462.2. [Example]

[0167] 1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)-1H-pyrrole-2,5-dione (IX) ZnCl (1.36 g, 10.0 mmol, 1 equiv.) was added to a 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 equiv.) in dry benzene (100 mL) and the mixture was heated to 80° C. Then, a solution of HMDS (2.42 g, 3.14 mL, 15.0 mmol, 1.5 equiv.) in dry benzene (10 mL) was added dropwise over 30 min. The reaction was continued to stir under reflux for approximately 24 h, then cooled and concentrated under reduced pressure. After evaporation of 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 NaSO and then evaporated to dryness. The crude product was purified by column chromatography using a DCM:MeOH (9:0.3; v / v) mixture as the eluent. The compound was obtained as a solid after washing with EtO.

[0168] White solid. Yield: 79% (3.34 g); 23 H 22 F3N3O4 (443.43), monoisotopic mass: 443.15. UPLC (purity = 99%): R =7.45 min. (M+H) + 444.1. [Example]

[0169] 3-(Methylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride A 2 M solution of methylamine in THF (0.07 g, 2.2 mmol, 1 equiv.) 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 equiv.) in dry benzene (50 mL). The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system. The compound was then converted to the hydrochloride salt by treating it with a 2 M methanolic hydrochloric acid solution.

[0170] White solid. Yield: 87% (0.91 g); mp 161.2-163.4°C; 24 H 25 F3N4O3 (474.48), monoisotopic mass: 474.19. UPLC (purity >99.9%): R = 5.53 min, (M+H) + 475.3. 1 H NMR (500 MHz, CDCl3) 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]

[0171] 3-(Dimethylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)-piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride The compound was prepared according to the 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 equivalent) and dimethylamine (0.10 g, 2.2 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system. The compound was converted to the hydrochloride salt by treating the compound with 2 M methanolic hydrochloric acid solution.

[0172] White solid. Yield: 83% (0.90 g); mp 157.8-159.2°C; 25 H 27 F3N4O3 (488.51), monoisotopic mass: 488.20. UPLC (purity >99.9%): R = 5.53 min, (M+H) + 489.3. 1 H NMR (500 MHz, CDCl3) 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). 13C NMR (126 MHz, CDCl3) 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]

[0173] 3-(Diethylamino)-1-(2-oxo-1-phenyl-2-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride The compound was prepared according to the 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 equivalent) and diethylamine (0.16 g, 2.2 mmol, 1 equivalent) were used as starting materials. The crude product was purified by column chromatography using a DCM:MeOH (9:0.5; v / v) elution system. The compound was converted to the hydrochloride salt by treating the compound with 2 M methanolic hydrochloric acid solution.

[0174] White solid. Yield: 88% (1.00 g); mp 142.2-143.1 °C; TLC: R f = 0.52 (DCM:MeOH (9:0.5; v / v)); C 27 H 31 F3N4O3 (516.57), monoisotopic mass: 516.23. UPLC (purity >99.9%): R = 5.79 min, (M+H) + 517.2. 1H NMR (500 MHz, DMSO-d6) 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]

[0175] Special properties of the water-soluble salts of the compounds according to the invention. 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 evaluated according to the methods described above, and the results are summarized in Tables 3 and 4.

[0176] [Table 5]

[0177] [Table 6]

[0178] Based on the results obtained, it was found that the salts of the compounds according to the invention clearly exhibit improved water solubility, which has a positive effect on their pharmacokinetic and / or pharmaceutical properties and is particularly advantageous in the case of intravenous administration of the compounds according to the invention.

Claims

1. General formula (I) 【Chemistry 1】 (In the formula, X is N or C; k is a number equal to 0 or 1; A is, - phenyl substituents; - halogen atom, -SCF ３ , -CF ３ , -CHF ２ , -CN, -OCF ３ , -NO ２ , -OCH ３ , -OC ２ H ５ a phenyl substituent substituted with one or two or three or four side substituents selected from the group consisting of straight or branched chain alkyl moieties having from 1 to 4 carbon atoms in the carbon backbone; a phenyl substituent, which is substituted with at least one aromatic or heteroaromatic substituent; - benzhydryl substituents; - 1-naphthyl or 2-naphthyl substituents; a benzothiophenyl substituent selected from the group consisting of a 2-benzothiophenyl, a 3-benzothiophenyl, a 4-benzothiophenyl, or a 5-benzothiophenyl substituent, preferably a 5-benzothiophenyl substituent; - benzisoxazole substituents selected from the group consisting of 3-benzisoxazole, 4-benzisoxazole, 5-benzisoxazole, 6-benzisoxazole, 7-benzisoxazole substituents, preferably the 5-benzisoxazole substituent; - a linear, branched or cyclic alkyl moiety having from 1 to 4 carbon atoms in the carbon backbone, preferably substituted with at least one halogen atom; is a substituent selected from the group consisting of: B is, - phenyl substituents; - halogen atom, -SCF ３ , -CF ３ , -CHF ２ , -CN, -OCF ３ , -NO ２ , -OCH ３ , -OC ２ H ５ , a straight or branched chain alkyl moiety having from 1 to 4 carbon atoms in the carbon backbone, and D is H, amino (-NH ２ ), one or two aliphatic substituents (especially -CH ３ and / or -C ２ H ５ or an amino group that is part of a heterocycle, or a pharma- ceutically acceptable salt thereof.

2. General formula (II) 【Chemistry 2】 (In the formula, X is N or C; k is a number equal to 0 or 1; A is, - phenyl substituents; - halogen atom, -SCF ３ , -CF ３ , -CHF ２ , -CN, -OCF ３ , -NO ２ , -OCH ３ , -OC ２ H ５ a phenyl substituent substituted with one or two or three or four side substituents selected from the group consisting of straight or branched chain alkyl moieties having from 1 to 4 carbon atoms in the carbon backbone; a phenyl substituent, which is substituted with at least one aromatic or heteroaromatic substituent; - benzhydryl substituents; - 1-naphthyl or 2-naphthyl substituents; a benzothiophenyl substituent selected from the group consisting of a 2-benzothiophenyl, a 3-benzothiophenyl, a 4-benzothiophenyl, or a 5-benzothiophenyl substituent, preferably a 5-benzothiophenyl substituent; - benzisoxazole substituents selected from the group consisting of 3-benzisoxazole, 4-benzisoxazole, 5-benzisoxazole, 6-benzisoxazole, 7-benzisoxazole substituents, preferably the 5-benzisoxazole substituent; - a linear or branched alkyl moiety having from 1 to 4 carbon atoms in the carbon backbone, preferably substituted with at least one halogen atom; is a substituent selected from the group consisting of: B is, - phenyl substituents; - halogen atom, -SCF ３ , -CF ３ , -CHF ２ , -CN, -OCF ３ , -NO ２ , -OCH ３ , -OC ２ H ５ , a straight or branched chain alkyl moiety having from 1 to 4 carbon atoms in the carbon backbone, (It is) 2. The compound according to claim 1, characterized in that it is a compound of the formula:

3. 3. The compound according to claim 1, wherein the halogen atom is a fluorine or chlorine atom.

4. 4. The compound according to any one of claims 1 to 3, characterized in that the straight or branched alkyl moiety in the carbon backbone contains from 1 to 4 carbon atoms, said alkyl moiety being selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl.

5. 5. A compound according to claim 1, characterized in that k=0.

6. 6. A compound according to claim 1, characterized in that X is nitrogen.

7. A compound according to any one of claims 1 to 6, characterized in that the substituent A is selected from the group consisting of 5-benzothiophenyl, 2-naphthyl, 5-benzisoxazolyl substituents.

8. The substituent A is phenyl; at least one chlorine atom or -CF ３ , -CHF ２ , -OCF ３ , -CH ３ , -SCF ３ 8. A compound according to claim 1, characterized in that it is selected from the group consisting of: or phenyl substituted with phenyl.

9. 9. A compound according to any one of claims 1 to 8, characterized in that the substituent B is selected from the group consisting of phenyl or phenyl substituted with one or two halogen atoms.

10. 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-yl)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, 10. A compound according to any one of claims 1 to 9, characterized in that it is selected from the group consisting of:

12. (R) enantiomer, preferably the following compound: (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 10. The compound according to claim 1, characterized in that it is selected from:

13. Water-soluble salts, especially hydrochlorides, preferably 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-(trifluoromethyl)phenyl)piperazin-1-yl)ethyl)pyrrolidine-2,5-dione hydrochloride 10. The compound according to claim 1, characterized in that it is selected from:

14. 14. A compound according to any one of claims 1 to 13 for use in the treatment or prevention of epileptic seizures, neuropathic pain, or migraine.

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