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    • 专利摘要:
    New compounds with antioxidant capacity that combine the inhibition of the monoaminoxidases and cholinesterase enzymes and the interaction with the histamine 3 receptor, its obtaining procedure and pharmaceutical compositions containing them. The present invention relates to new compounds with antioxidant capacity that combine the inhibition of the monoaminoxidases and cholinesterase enzymes and the interaction with the histamine H3 receptor, the process for its preparation, the pharmaceutical compositions containing the new derivatives and their use in the treatment of neurodegenerative and central nervous system diseases. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2701954A1
    申请号:ES201731044
    申请日:2017-08-23
    公开日:2019-02-26
    发明作者:Muñoz Francisco López；Contelles José Luis Marco；Holger Stark；Stefanie Hagenow；Rona R Ramsay
    申请人:Agencia Estatal Consejo Superior De Investig Cientificas M P；Centro De Ensenanza Univ Sek S A；
    IPC主号:
    专利说明:

    [0001]
    [0002] New compounds with antioxidant capacity that combine the inhibition of monoaminoxidases and cholinesterase enzymes and the interaction with the histamine 3 receptor, its obtaining procedure and pharmaceutical compositions containing them
    [0003]
    [0004] The present invention relates to new compounds with antioxidant capacity that combine the inhibition of the monoaminoxidases and cholinesterase enzymes and the interaction with the histamine H3 receptor, the process for obtaining them, the pharmaceutical compositions containing the new derivatives and their use in the treatment of neurodegenerative and central nervous system diseases.
    [0005]
    [0006] More specifically, the invention describes the preparation and study as multidiana inhibitors of the enzymes monoaminooxidasas A and B, acetylcholinesterase and butyrylcholinesterase, and antagonists of the histamine H3 receptor, with antioxidant capacity, of a new derivative of W-methyl-W - {[ (1-methyl-5-alkoxy) -1H-indol-2-yl] methyl} prop-2-ino-1-amine, as well as its application within the field of the pharmaceutical industry, for the production of drugs for the purpose of curing, stopping or alleviating diseases of the central nervous system and neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, stroke, amyotrophic lateral sclerosis, narcolepsy, and disorders and sleep disturbances.
    [0007]
    [0008] Thus, in a first aspect, the invention provides new compounds with antioxidant capacity that combine the inhibition of the monoaminoxidases and cholinesterase enzymes and the interaction with the histamine H3 receptor. Likewise, in a second aspect, the invention provides a process for the preparation of said compounds. In a third aspect, the invention relates to the use of these derivatives for the preparation of medicaments for the purpose of curing, stopping or mitigating diseases of the central nervous system and neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, stroke, sclerosis. lateral amyotrophic, narcolepsy and disorders and sleep disturbances.
    [0009] Among the neurodegenerative diseases, Alzheimer's disease (AD) is the most common form and cause of memory loss and other cognitive defects, such as disorientation, depression, inability to perform the most common and daily tasks of daily life, which They are diagnosed among the elderly and members of the so-called third age (Querfurth, HW, LaFerla, FMN Engl. J. Med. 2010, 362, 329-344).
    [0010]
    [0011] The EA is characterized by a series of highly interconnected pathological processes that are translated, from the biological and biochemical point of view, in the accumulation and aggregation of an abnormal amount of extracellular deposits of beta-amyloid peptide (Ap) that cause senile plaques , the formation of intracellular neurofibrillary tangles, composed of tauhiperphosphorylated protein, and which, finally, trigger neuronal death processes (Bartus, RT, Dean, RL, Beer, B., Lippa, AS Science 1982, 217, 408-414).
    [0012] In the search to know and understand the mechanisms that lead to neurodegeneration, and, therefore, to design and find new and more efficient therapies for its treatment, factors such as mitochondrial dysfunction, neuroinflammation, and oxidative stress (von Bernhardi, R., Eugenín, J. Antioxid, Redox Signal, 2011, 16, 974-1031) have been identified as the main and most determining factors in the progress and development of AD. Indeed, it has been shown that the presence and formation of senile plaques leads to an increase in the number and toxic activity of reactive oxygen species (ROS) that cause damage to the nucleus, at the mitochondrial level and acting on cytoplasmic proteins. neuronal That is why the antioxidant strategy proposed in the development of new drugs for age-related diseases, such as AD, is considered of the utmost importance and interest (Rosini, M .; Simoni, E.; Milelli, A.; Minarini , A., Melchiorre, CJ Med. Chem. 2014, 57, 2821-2831).
    [0013]
    [0014] The therapy that is currently administered to patients with AD is basically symptomatic and palliative, which slows down its development, but does not cure it. Thus, in recent decades, the cholinergic approach has placed four drugs on the market for the treatment of the disease, characterized as being inhibitors of the enzyme acetylcholinesterase (AChE), increasing neurotransmission in the cholinergic synapses of the brain, and palliating the cognitive deficit, by preventing hydrolysis, and consequently, restoring the level of the neurotransmitter acetylcholine (Villarroya, M. et al., Expert Opin, Investig Drugs 2007, 16, 1987-1998). In addition to its function in cholinergic transmission, AChE has other functions related to neuronal differentiation, cell adhesion and Ap aggregation. Different studies have shown that AChE favors the formation of Ap aggregates, giving rise to complex AChE-Ap which are more toxic than the isolated Ap itself. Since the point of adhesion between the enzyme and the peptide is located in the peripheral site of the enzyme (PAS). Therefore, the dual AChE inhibitors (Fernández-Bachiller, MI et al., ChemMedChem 2009, 4, 828-841; Muñoz-Torrero, D. Curr. Med. Chem. 2008, 15, 2433-2455), able to interact simultaneously in the central catalytic site and the PAS, they would be of great interest in the therapy of AD, since they can alleviate the cognitive deficiencies and stop the neurotoxicity related to Ap (de Ferrari, GV et al., Biochemistry 2001, 40, 10447 -10457).
    [0015]
    [0016] The inhibition of monoamine oxidase (MAO) enzymes has been proposed as an interesting pharmacological target for the design of new drugs and approach of the possible treatment of AD and other neurodegenerative diseases, such as Parkinson's disease, since during the deamination reaction of the neurotransmitter amines, such as adrenaline, dopamine and serotonin, catalyzed by the MAOs, generates hydrogen peroxide (H 2 O 2 ), which is a source of ROS, agents responsible for oxidative stress and which very negatively affect neurons in the beginning and course of these pathologies (Schneider, LS et al., Am. J. Psychiatry 1993, 18, 321-323; Marin, DB et al., Psychiatry Res. 1995, 58, 181 189; Alper, G. et al. Eur. Nueropsychopharmacol. 1999, 9, 247-252).
    [0017]
    [0018] Another interesting strategy known and proposed for the design of new drugs for the treatment of AD is the search and identification of histamine H3 receptor antagonists (H3R). H3R is predominantly expressed in the central nervous system (CNS), where it functions as an auto-receptor, modulating the release of histamine, and as a hetero-receptor, regulating the release of multiple neurotransmitters. That is why the H3R antagonists are potential therapeutic agents for the treatment of cognitive deficits associated with various pathologies, among which is the EA (tazewska, D., Kiec-Kononowicz, K. Expert Opin , Ther.Patents 2014, 24, 89 111; Saad, A., Sadeq, A., Jalal, F., Stark, H. Behav, Brain Res. 2016, 312, 415-430).
    [0019]
    [0020] Since H3R is involved in the central regulation of histamine levels (Walter, M., Stark, H. Front, Biosci, Schol, Ed. 2012, 4, 461-488), and other neurotransmitters (serotonin, dopamine and norepinephrine) ) (Ellenbroek, BA, Ghiabi, B. Trends in Neurosciences 2014, 37, 191-199), is considered as a pharmacological tool of first order for the development of drugs for CNS diseases, as is the case of the antagonist H3R pitolisant (WAKIX®), approved in 2016 by the European Medicines Agency for the treatment of narcolepsy, and in development and research for the treatment of sleep and other cognitive disorders (Schlicker, E., Kathmann, N. In The Histamine H3 Receptor Leurs, R., Timmerman, H., Eds., A target for new drugs, Elsevier Science BV, 1998, pp 13-26).
    [0021]
    [0022] In this context, propargylamine ASS234 (Bolea, I. et al., J. Med. Chem. 2011, 54, 8251-8270) has been described in the literature (Figure 1), as a molecule capable of inhibiting MAOs, preventing catalytic oxidation of biogenic amines, deficient in the brains of patients with AD, and to avoid the formation of H 2 O 2 , the starting point for the genesis of ROS via Fenton and Haber-Weiss reaction in the presence of biometals ( Zheng, H. et al., J. Neurochem , 2005, 95, 68 78). On the other hand, the ASS234 molecule is a reversible inhibitor of the mixed, potent and selective type of AChE.
    [0023]
    [0024] Although compounds are known to occur as antagonists of H3R and inhibitors of AChE (Petroianu G, Arafat K, Sasse BC, Stark H. Pharmazie 2006, 61, 179-182; Bembenek, SD et al., Bioorg, Med. Chem. 2008 , 16, 2968-2973; Morini, G. et al., Bioorg, Med. Chem. 2008, 16, 9911-9924; Darras, FH et al., ACS Chem. Neurosci., 2014, 5, 225-242; Bajda, M; et al., Arch. Pharm. 2012, 345, 591-597), or H3R antagonists and inhibitors of Ap aggregation, chelators of metals and "scavengers" of ROS (Sheng, R. et al., ACS Chem. Neurosci 2016, 7, 69-81), which is absolutely novel and original and until now not described in the literature, and will be the object of the present patent, are the multidiana molecules (León, R., Garcia, AG, Marco-Contelles, J. Med. Res. Rev. 2013, 33, 139-189) of the tripotent type capable of modulating the H3R receptor, showing ability to inhibit MAO and ChEs, with antioxidant capacity, which is an original therapeutic approach for the development of molecules for the potential treatment of CNS diseases, and neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, stroke, amyotrophic lateral sclerosis , narcolepsy and alterations and sleep disorders.
    [0025]
    [0026] Figure 1 shows an illustrative example of the design that has been used for some of the possible new products of the invention of type A, from the ASS234 molecule, after simple structural modification and functional, judicious and rational, has been incorporated the antagonist pharmacophore of H3R plus the ChE motif, composed of a tertiary amine in the western part of the molecule, connected with a spacer to the eastern part of the molecule, where the MA-motif of propargylamine is located. In fact, cycloalkylamines, such as piperidine, connected via a (propyloxy) phenyl chain, to an arbitrary and flexible region in the eastern part of the molecule, function as well-known and exploited H3R antagonistic motifs, as in pitolisant, irdabisant or JNJ-5207852 (Wingen, K .; Stark, H. Drug Disc., Today Techn., 2013, 10, e483-e489; Celanire, S. et al., Drug Discov Today , 2005, 10, 1613-27).
    [0027]
    [0028] In a first aspect, the invention provides novel derivatives N-methyl-N - {[(1-methyl-5-alkoxy) -1H-indol-2-yl] methyl} prop-2-yne-1-amine with antioxidant capacity which combine the inhibition of the monoaminoxidases and cholinesterase enzymes and the interaction with the histamine H3 receptor, and which, therefore, are multipotent inhibitors of the monoamine oxidase enzymes A and B, acetylcholinesterase and butyrylcholinesterase and histamine H3 receptor antagonists,
    [0029]
    [0030] Thus, according to this first aspect of the invention, the invention relates to a compound of formula (I)
    [0031]
    [0032]
    [0033] where,
    [0034]
    [0035] the symbol () m represents a linear alkyl chain composed of m -CH 2 groups, m may vary between 0 and 6;
    [0036]
    [0037] the symbol () n represents a group -CH 2 , being able to vary n between 0 and 6, of form
    [0038]
    [0039]
    [0040]
    [0041] that the cyclic part v 1 of the compound of formula (I) can be constituted by a cycle of 4 ring members (n = 0), of 5 ring members (n = 1), of 6 ring members (n = 2) ) and so on up to 10 ring members (n = 6);
    [0042] R1 and R2 are selected, independently from each other, from H or straight or branched, substituted or unsubstituted (C 1 -C 10 ) alkyl;
    [0043]
    [0044] R3 is selected from H, -OR4, N, -CN, -C (O) R4, -C (O) OR4, -C (O) NR4R5,
    [0045] -C = NR 4, -OC (O) R 4, -NR 4 R 5, -NR 4C (O) R 5, -NO 2 , -N = CR 4 R 5, halogen or (C 1 -C 10 ) alkyl, with R 4 and R 5 being selected from H, alkyl linear or branched, substituted or unsubstituted, linear or branched alkenyl, substituted or unsubstituted, substituted or unsubstituted cycloalkyl and substituted or unsubstituted aryl (heteroaryl);
    [0046]
    [0047] X is CH 2 , O, S, Se, NR 6, R 6 being selected from H, linear or branched alkyl, substituted or unsubstituted, linear or branched alkenyl, substituted or unsubstituted, substituted or unsubstituted cycloalkyl and substituted or unsubstituted aryl (heteroaryl);
    [0048]
    [0049] optionally in the form of a simple stereoisomer or a mixture of stereoisomers, in the form of free compound and / or physiologically acceptable salt and / or a physiologically acceptable solvate thereof.
    [0050]
    [0051] For the purposes of the present invention, the term "simple stereoisomer" preferably means a single enantiomer or diastereomer, and in the sense of this invention, the term "mixture of stereoisomers" means a racemate and mixtures of enantiomers and / or diastereomers, in any mixing ratio.
    [0052]
    [0053] In the sense of this invention, the term "physiologically acceptable salt" comprises a salt of at least one compound according to the present invention and at least one physiologically acceptable acid or base.
    [0054]
    [0055] In the sense of this invention, a physiologically acceptable salt of at least one compound according to the present invention and at least one physiologically acceptable acid preferably refers to a salt of at least one compound according to the present invention with at least one Inorganic or organic acid that is physiologically acceptable, in particular if it is used in humans and / or other mammals. Examples of physiologically acceptable acids are hydrochloric, hydrobromic, sulfuric, methanesulfonic, p-toluenesulfonic, carbonic, formic, acetic, oxalic, succinic, tartaric, mandelic, fumaric, maleic, lactic, citric, glutamic, saccharic, monomethyl-sebacic, 5- oxoproline, hexane-1-sulphonic acid, nicotinic acid, 2, 3 or 4-aminobenzoic acid, 2,4,6-trimethylbenzoic acid, α-lipoic acid, acetylglycine, hippuric acid, phosphoric acid, aspartic acid, with preference being given to oxalic and hydrochloric acids, as well as the corresponding mono- or bis-oxalates or hydrochlorides.
    [0056]
    [0057] In the sense of this invention, a physiologically acceptable salt of at least one compound according to the present invention and at least one physiologically acceptable base preferably refers to a salt of at least one compound according to the present invention, as an anion, with at least one preferably inorganic cation which is physiologically acceptable, in particular if it is used in humans and / or other mammals.
    [0058] For the purposes of this invention, the term "physiologically acceptable solvate" preferably comprises an adduct of a compound according to the present invention and / or a physiologically acceptable salt of at least one compound according to the present invention with different molecular equivalents of one or more solvents. Examples of physiologically acceptable solvents are: water, alkanols, esters, ethers or ketones. Hydrates are particularly preferred solvates.
    [0059]
    [0060] In the present description, the term "alkyl" refers to straight or branched hydrocarbon chain residues, having from 1 to 10 carbon atoms, preferably from 1 to 4, and which are attached to the rest of the molecule via a bond simple, for example methyl, ethyl, n-propyl, / -propyl, n-butyl, fer-butyl, sec-butyl, n-pentyl, n-hexyl, etc. The alkyl groups may be optionally substituted with one or more substituents selected, independently from each other, from halogen, hydroxyl, alkoxy, carboxyl, carbonyl, cyano, acyl, alkoxycarbonyl, amino, nitro, mercapto and alkylthio, wherein the alkyl groups of these substituents are selected from linear or branched alkyl, substituted or unsubstituted, linear or branched alkenyl, substituted or unsubstituted, substituted or unsubstituted cycloalkyl and substituted or unsubstituted aryl or heteroaryl;
    [0061]
    [0062] In the present description, the term "halogen" refers to any halogen F, Cl, Br and I.
    [0063]
    [0064] In the present description, the term "cycloalkyl" refers to each of the radicals derived from cycloalkanes, of general formula CfH 2 M, where t is from 1 to 10, preferably from 1 to 4.
    [0065]
    [0066] In the present description, the term "alkenyl" is a group derived from an alkene of 2 to 10 carbon atoms, preferably of 2 to 4 carbon atoms, which includes a double bond C = C by removal of one of its carbon atoms. hydrogen.
    [0067]
    [0068] In the present description, the term "aryl" refers to an aromatic hydrocarbon derivative, such as, without being limited to, phenyl, naphthyl, indenyl, phenanthryl or anthracyl, and the term "heteroaryl" refers to a cyclic aromatic group of 5 or 6 members containing at least 1 and if appropriate also 2, 3, 4 or 5 heteroatoms, the heteroatoms being selected, in each case independently of each other, from the group consisting of S, N and O.
    [0069]
    [0070] In a preferred embodiment of the compound of formula (I) R3 is H.
    [0071]
    [0072] In another preferred embodiment, R1 and R2 are, independently of each other, a linear or branched, substituted or unsubstituted (C 1 -C 4 ) alkyl group.
    [0073]
    [0074] In another preferred embodiment of the compounds of the invention, they are represented by the following formula (II), wherein R3 is H, X is CH 2 and R1 and R2 are both a methyl group and m and n are an integer from 0 to 6 such as defined above for the compounds of formula (I)
    [0075]
    [0076]
    [0077]
    [0078]
    [0079] Particularly preferred are compounds of formula (II) selected from among
    [0080]
    [0081] • W-Methyl-N - ((1-methyl-5- (2- (pyrrolidin-1-yl) ethoxy) -1 H -indol-2-yl) methyl) prop-2-yn-1 -amine
    [0082] • W-Methyl-W - ((1-methyl-5- (3- (pyrrolidin-1-yl) propoxy) -1 H -indol-2-yl) methyl) prop-2-yn-1-amine
    [0083] • W-Methyl-W - ((1-methyl-5- (2- (piperidin-1-yl) ethoxy) -1 H -indol-2-yl) methyl) prop-2-in-1 -amine
    [0084] • W-Methyl-W - ((1-methyl-5- (3- (piperidin-1-yl) propoxy) -1 H -indol-2-yl) methyl) prop-2-yn-1-amine
    [0085] • W-Methyl-W - ((1-methyl-5 - ((5- (piperidin-1 -yl) pentyl) oxy) -1 H -indol-2-yl) methyl) prop-2-in-1- amine.
    [0086] Particularly preferred is the compound of formula (II) which is N-Methyl-N - ((1-methyl-5- (3- (piperidin-1-yl) propoxy) -1H-indol-2-yl) methyl) prop -2-in-1 -amine.
    [0087] The compounds of the present invention represented by the formula (I) may include isomers, depending on the presence of multiple bonds (eg, Z, E), including optical isomers or enantiomers, depending on the presence of chiral centers. Individual isomers, enantiomers or diastereomers and mixtures thereof fall within the scope of the present invention, i.e., the term isomer also refers to any mixture of isomers, such as diastereomers, racemates, etc., even to their optically isomers active or mixtures in different proportions thereof. The individual enantiomers or diastereomers, as well as their mixtures, can be separated by conventional techniques.
    [0088]
    [0089] The compounds according to the first aspect of the present invention and the corresponding stereoisomers and the respective salts and solvates are toxicologically safe and, consequently, are suitable as pharmacologically active ingredients in pharmaceutical compositions.
    [0090]
    [0091] Thus, in another aspect of the present invention, the invention also relates to a pharmaceutical composition comprising at least one compound of formula (I) as defined above and optionally at least one pharmaceutically acceptable adjuvant, excipient and / or carrier and / or, if appropriate, one or more additional pharmacologically active compounds.
    [0092]
    [0093] The pharmaceutical composition according to the invention is suitable to be administered to adults and children, including toddlers and infants.
    [0094]
    [0095] The pharmaceutical composition according to the invention can be prepared as a liquid, semisolid or solid pharmaceutical form, for example in the form of solutions for injection, drops, juices, syrups, sprays, suspensions, tablets, patches, capsules, dressings, suppositories, ointments, creams, lotions, gels, emulsions, aerosols or in multiparticulate form, for example in the form of pills or granules, if appropriate compressed into tablets, decanted into capsules or suspended in a liquid, and administered as such.
    [0096] In addition to at least one compound according to the invention, if appropriate in the form of one of its pure stereoisomers, in particular enantiomers or diastereoisomers, in the form of its racemate or in the form of mixtures of the stereoisomers, in particular of enantiomers or diastereomers , in any desired mixing ratio, or if appropriate in the form of a corresponding salt, or respectively in the form of a corresponding solvate, the pharmaceutical composition according to the invention can conventionally contain other physiologically acceptable pharmaceutical adjuvants.
    [0097] The pharmaceutically acceptable adjuvants and vehicles which may be used in said compositions are the adjuvants and vehicles known to those skilled in the art and commonly used in the preparation of therapeutic compositions, which may be selected, for example, from the group consisting of excipients, fillers, solvents, diluents, surfactants, dyes, preservatives, disintegrants, glidants, lubricants, flavorings and binders.
    [0098]
    [0099] The selection of physiologically compatible adjuvants and the amount to be used thereof depend on the form of administration of the pharmaceutical composition, ie orally, subcutaneously, parenterally, intravenously, intraperitoneally, intradermally, intramuscularly, intranasally, buccally or rectally. . The preparations in the form of tablets, dragees, capsules, granules, pills, drops, juices and syrups are preferably suitable for oral administration; the solutions, suspensions, dry preparations which are easily reconstituted and also sprays are preferably suitable for parenteral, topical and inhalation administration. The compounds according to the invention used in the pharmaceutical composition according to the invention in a tank, in a dissolved form or in a dressing, and if appropriate having added other agents that favor penetration into the skin, are suitable preparations for percutaneous administration. The orally or percutaneously administrable preparation forms can also release the respective compound according to the invention in a delayed manner.
    [0100] The pharmaceutical compositions according to the invention are prepared with the aid of conventional means, devices, methods and processes known in the art, such as are described, for example, in "Remington's Pharmaceutical Sciences", AR Gennaro (Editor), Edition 17, Mack Publishing Company, Easton, Pa., 1985, in particular in volume 8, chapters 76 to 93.
    [0101]
    [0102] In a particular embodiment, said pharmaceutical composition is prepared in the form of a solid form or aqueous suspension, in a pharmaceutically acceptable diluent, preferably for oral, topical, rectal or parenteral administration (including subcutaneous, intraperitoneal, intradermal, intramuscular, intravenous, etc.).
    [0103]
    [0104] For their application in therapy, the compounds of formula (I), salts or isomers thereof will be found, preferably, in a pharmaceutically acceptable or substantially pure form, ie, with a pharmaceutically acceptable level of purity excluding normal pharmaceutical additives such as diluents and carriers, and not including material considered toxic at normal dosage levels. The purity levels for the active ingredient are preferably greater than 50%, more preferably greater than 70%, and still more preferably greater than 90%. In a preferred embodiment, they are greater than 95% of the compound of formula (I).
    [0105]
    [0106] The amount of the respective compounds according to the invention of the general formula (I) shown above to be administered to patients is variable and depends for example on the weight or age of the patient and also on the type of administration, the indication and the severity of the illness. Normally they are administered between 0.001 and 100 mg / kg of body weight of the patient,
    [0107]
    [0108] In a particularly preferred embodiment, the compounds of the invention are administered in the form of mono- or bis-oxalate salts or hydrochloride of the corresponding derivatives, in doses of 6.0 mg / kg to 10.00 mg / kg of body weight of the patient, in particular in doses of 6.7 mg / Kg to 9.3 mg / Kg.
    [0109] In another aspect, the present invention relates to the use of a compound of formula (I) as described above for the manufacture of a medicament.
    [0110]
    [0111] In another aspect, the present invention relates to the use of a compound of formula (I) as described above for the manufacture of a medicament for the treatment of a neurodegenerative or central nervous system disease.
    [0112] In a preferred embodiment, the neurodegenerative or central nervous system disease is selected from senile dementia, cerebrovascular dementia, mild cognitive deficit, attention deficit disorders, neurodegenerative diseases associated with aberrant protein aggregations such as Parkinson's disease or Alzheimer's disease. , amyotrophic lateral sclerosis, prion diseases such as Creutzfeldt-Jakob disease or Gerstmann-Straussler-Scheinker disease, stroke, amyotrophic lateral sclerosis, narcolepsy and disorders and sleep disturbances. In a particularly preferred embodiment, the neurodegenerative disease is Parkinson's disease or Alzheimer's disease.
    [0113]
    [0114] In another aspect, the present invention relates to a process for obtaining the compounds of formula (I), which comprises the reaction of a compound of formula (III)
    [0115]
    [0116]
    [0117]
    [0118] where R1, R2 are defined as above
    [0119]
    [0120] with a compound of formula (IV):
    [0121]
    [0122]
    [0123] where R3, X, n and m are defined as above,
    [0124]
    [0125] the reaction being carried out in a polar solvent such as anhydrous dimethylformamide, under an argon atmosphere, with the addition of sodium hydride.
    [0126]
    [0127] General synthesis of the compounds of formula ( I)
    [0128]
    [0129] To a solution of the corresponding compound of formula (III) (1 mmol) and of the corresponding cycloalkylamine hydrochloride of formula (IV) (1 eq.) In dry dimethylformamide (DMF) (0.08 M), under argon, NaH was added. (3 eq., 60% dispersion in mineral oil) slowly. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure and an aqueous saturated solution of NH 4 Cl was added, and it was extracted with EtOAc several times. The combined organic phase was washed with a saturated solution of sodium chloride, dried over Na 2 SO 4 , filtered, and the solvent was evaporated to dryness, under reduced pressure. The resulting crude was purified by column chromatography (hexane / EtOAc, 10-50%) to obtain the desired compounds of formula (I).
    [0130]
    [0131] Examples
    [0132]
    [0133] The invention is illustrated below by means of examples and tests that show the specificity and effectiveness of the compounds of formula (I) of the invention.
    [0134]
    [0135] All the anhydrous solvents were distilled using a Pure solv solvent purification system model PS-400-3-MD. The melting points (are not corrected), and have been measured in a Kofler type microscope (Reicher Jung Thermovar). The 1 H NMR and 13 C NMR spectra were performed on a Varian Inova-300 (300 MHz), Mercury-400 (400 MHz), Varian Inova-400 (400 MHz) and Unity-500 (500 MHz). The chemical shifts (in ppm) have been referenced to the residual signal of the solvent used [CDCl3: 7.27 (D), 77.2 (C) ppm; CD3OD: 4.84 (D), 49.05 (C) ppm]. The multiplicity of signals: s, singlet; d, doublet; t, triplet; c: quadruplet, q, quintuplet; m, multiplet), br means width, the number of protons (deduced by integration), the value of the coupling constants J (in hertz) and the structural assignment, inferred from the study of two-dimensional experiments (1H, 1H-COZY, 1H , 13C-HSQC, 1H, 13C-HMBC). The mass spectra were recorded on an HP-1100MSD LC / MS spectrometer with APCI and API-ES ionization sources. Also, in the indicated cases, the spectra were recorded by electronic impact in an HP-5873MSD spectrometer for sample injection by direct probe. The infrared spectra were acquired in a Perkin-Elmer Spectrum One apparatus in KBr pellet. The most significant bands are indicated in cm-1. The elemental analyzes have been carried out with a Heraus CHN-O Rapid analyzer and are expressed in percent. The chromatographic separations were carried out by column chromatography using Merck 60 (0.063-0200 nm) silica gel under pressure (flash) and gradient, using as eluent the detailed mixtures in each case, or by chromatotron (accelerated centrifugal radial chromatography) model 7924 with Merck silica gel plates 60 F 254 - 366 . For the thin layer chromatography PL chromatofoliums have been used. of Merck F244 silica gel.
    [0136]
    [0137] Example 1: Synthesis of W-Methyl-W - ((1-methyl-5- (2- (pyrrolidm-1-yl) ethoxy) -1H-mdol-2-yl) methyl) -prop-2-in-1 -amine
    [0138]
    [0139] Following the general synthesis of the compounds of formula (I), to a solution of 1-methyl-2 - ((methyl (prop-2-in-1-yl) amino) methyl) -1H-indol-5-ol ( crosses, MA; Elorriaga, C .; Fernandez-Alvarez, E. Acetylenic and allenic derivatives of 2- (5-benzyloxyindolyl) and 2- (5-hydroxyindolyl) methylamines: synthesis and in vitro evaluation as monoamine oxidase inhibitors Eur J.. Med Chem. 1991 , 26, 33-41) (120 mg, 0.52 mmol) and 1- (2-chloroethyl) pyrrolidine hydrochloride (89.1 mg, 0.52 mmol) in dry DMF (7 ml. ), under argon, NaH (38 mg, 1.56 mmol, 60% dispersion in mineral oil) was added slowly. The reaction mixture was stirred at room temperature overnight. The solvent was evaporated under reduced pressure and added a saturated aqueous solution of NH 4 Cl (50 ml), and extracted with EtOAc several times (3x200 ml). The combined organic phase was washed with a saturated solution of sodium chloride, dried over Na 2 SO 4 , filtered, and the solvent was evaporated to dryness, under reduced pressure. The resulting crude was purified by column chromatography (hexane / EtOAc, 10-50%) to obtain the title compound (121 mg, 71%) isolated as a yellow oil:
    [0140]
    [0141] Rf = 0.39 (hexane / AcOEt, 70%);
    [0142]
    [0143] IR (KBr) v 3433, 2955, 2620, 2126, 1724, 1625, 1487, 1405, 1279, 1209 cm-1;
    [0144] 1 H-NMR (500 MHz, CDCb) 57.16 (d, J = 8.8 Hz, 1H), 7.03 (d, J = 2.4 Hz, 1H), 6.81 (dd, J = 8 , 8, 2.4 Hz, 1H), 6.32 (s, 1H), 4.49-4.47 (t, J = 4.9 Hz, 2H), 3.72 (s, 3H), 3 , 65 (s, 2H), 3.46 (J = 4.9 Hz, 2H), 3.49-3.44 (m, 4H), 3.28 (d, J = 2.4 Hz, 2H) , 2.32 (s, 3H), 2.28 (t, J = 2.4 Hz, 1H), 2.27-2.11 (m, 4H);
    [0145]
    [0146] 13 C-NMR (126 MHz, CDCb) 5 151.5, 137.6, 133.9, 127.5, 111.3, 109.8, 104.0, 102.24, 78.30, 73.5, 64.3, 54.1 (2 C), 53.9, 51.7, 44.7, 41.5, 29.9, 23.2 (2 C);
    [0147]
    [0148] MS (ESI) m / z: 326.3 (M + 1) +.
    [0149]
    [0150] The obtained compound was transformed into its bis-oxalate: melting point 183-5 ° C;
    [0151] 1 H-NMR (300 MHz, D 2 O) 5 7.33 (d, J = 9.0 Hz, 1H), 7.12 (d, J = 2.4 Hz, 1H), 6.94 (dd, J = 9.0, 2.2 Hz, 1H), 6.67 (s, 1H), 4.55 (br s, 2H), 4.24 (t, J = 4.9 Hz, 2H), 3 , 93 (d, J = 1.9 Hz, 2H), 3.65 (s, 3H), 3.62-3.56 (m, 2H), 3.51 (t, J = 4.9 Hz, 2H), 3.09-3.03 (m, 3H), 2.84 (s, 3H), 2.04-2.02 (m, 2H), 1.91-1.87 (m, 2H) ,
    [0152]
    [0153] Analysis C 20 H 27 N 3 O 2 XHCO 2 CO 2 HH 2 O
    [0154] Calculated: C, 55.06; H, 6.35; N, 8.03
    [0155] Found: C, 54.97; H, 6.15; N, 7.50
    [0156]
    [0157] Example 2: Synthesis of W-Methyl-W - ((1-methyl-5- (3- (pyrrolidm-1-yl) propoxy) -1H-indol-2-yl) methyl) prop-2-in-1 - amine
    [0158]
    [0159] Following the general synthesis of the compounds of formula (I), from 1-methyl-2 - ((methyl (prop-2-yn-1-yl) amino) methyl) -1H-indol-5-ol (Crosses , MA; Elorriaga, C .; Fernández-Álvarez, E. Acetylenic and allenic derivatives of 2- (5-benzyloxyindolyl) and 2- (5-hydroxyindolyl) methylamines: synthesis and in vitro evaluation as monoamine oxidase inhibitors. Eur. J. Med. Chem. 1991 , 26, 33-41) (150 mg, 0.65 mmol) and 1- (3-chloropropyl) pyrrolidine hydrochloride (121 mg, 0.65 mmol) in dry DMF (10 mg). ml), by reaction with NaH (47 mg, 1.95 mmol, 60% dispersion in mineral oil), after column chromatography (hexane / EtOAc, 10-50%), the title compound was obtained (144 mg, 65%). as a yellow oil:
    [0160]
    [0161] Rf = 0.37 (hexane / AcOEt 70%);
    [0162]
    [0163] 1 H-NMR (300 MHz, CDC b) 57.19 (d, J = 8.8 Hz, 1H), 7.08 (d, J = 2.5 Hz, 1H), 6.90 (dd, J = 8 , 8, 2.5 Hz, 1H), 6.36 (s, 1H), 4.09 (t, J = 6.4 Hz, 2H), 3.75 (s, 3H), 3.69 (s) , 2H), 3.33 (d, J = 2.2 Hz, 2H), 2.74-2.71 (m, 2H), 2.69-2.59 (m, 4H), 2.37 ( s, 3H), 2.32 (t, J = 2.2 Hz, 1H), 2.11-2.08 (m, 2H), 1.85-1.80 (m, 4H);
    [0164]
    [0165] MS (ESI) m / z 340.3 (M + 1) +.
    [0166]
    [0167] The obtained compound was transformed into its bis-oxalate: melting point 156-9 ° C;
    [0168] IR (KBr) v 3432, 3263, 2958, 2618, 2127, 1723, 1624, 1487, 1406, 1279, 1207, 1104, 1058 cm-1;
    [0169]
    [0170] 1 H-NMR (500 MHz, D 2 O) 57.31 (d, J = 9.3 Hz, 1H), 7.10 (d, J = 2.4 Hz, 1H), 6.90 (dd, J = 9.3, 2.5 Hz, 1H), 6.65 (s, 1H), 4.52 (br s, 2H), 4.03 (t, J = 5.8 Hz, 2H), 3, 91 (d, J = 2.4 Hz, 2H), 3.63 (s, 3H), 3.57-3.52 (m, 2H), 3.28-3.25 (m, 2H), 3 , 03 (t, J = 2.4 Hz, 1H), 2.95-2.93 (m, 2H), 2.82 (s, 3H), 2.08-2.05 (m, 2H), 2.01-1.94 (m, 2H), 1.90-1.84 (m, 2H);
    [0171]
    [0172] 13 C-NMR (126 MHz, D 2 O) 5 167.8, 154.8, 136.4, 130.8, 129.2, 116.5, 114.0, 106.6, 82.8, 74, 0, 68.5, 56.7 (2 C), 55.0, 52.1.46.8, 41.8, 32.3, 27.8, 25.0 (2 C),
    [0173] Analysis C 21 H 29 N 3 O 2 XHCO 2 CO 2 HH 2 O
    [0174] Calculated: C, 55.86; H, 6.56; N, 7.82
    [0175] Found: C, 56.15; H, 6.44; N, 7.64
    [0176] Example 3: Synthesis of W-Methyl-W - ((1-methyl-5- (2- (piperidm-1-N) ethoxy) -1H-mdol-2-yl) methyl) -prop-2-in-1 -amine
    [0177]
    [0178] Following the general synthesis of the compounds of formula (I), from 1-methyl-2 - ((methyl (prop-2-yn-1-yl) amino) methyl) -1H-indol-5-ol (Crosses , MA, Elorriaga, C .; Fernández-Álvarez, E. Acetylenic and allenic derivatives of 2- (5-benzyloxyindolyl) and 2- (5-hydroxyindolyl) methylamines: synthesis and in vitro evaluation as monoamine oxidase inhibitors, Eur. J. Med. Chem. 1991 , 26, 33-41) (200 mg, 0.87 mmol) and 1- (2-doroethyl) piperidine hydrochloride (160 mg, 0.87 mmol) in dry DMF (10 mL), by reaction with NaH (63 mg, 2.61 mmol, 60% dispersion in mineral oil), after column chromatography (hexane / EtOAc, 10-50%), the title compound (170 mg, 60%) was obtained as a yellow oil
    [0179]
    [0180] Rf = 0.35 (hexane / EtOAc, 70%);
    [0181]
    [0182] 1 H-NMR (300 MHz, CDC b) 57.17 (d, J = 9.0 Hz, 1H), 7.05 (d, J = 2.2 Hz, 1H), 6.87 (dd, J = 9 , 0, 2.2 Hz, 1H), 6.34 (s, 1H), 4.17 (t, J = 6.1 Hz, 2H), 3.73 (s, 3H), 3.67 (s) , H), 3.31 (d, J = 2.4 Hz, 2H), 2.83 (t, J = 6.1 Hz, 2H), 2.57 (t, J = 5.0 Hz, 4H ), 2.34 (s, 3H), 2.30 (t, J = 2.4 Hz, 1H), 1.67-1.61 (m, 4H), 1.48-1.46 (m, 2H);
    [0183]
    [0184] MS (ESI) m / z 340.4 (M + 1) +,
    [0185]
    [0186] The obtained compound was transformed into its bis-oxalate: melting point 153-5 ° C;
    [0187] IR (KBr) v 3426, 3276, 2953, 2129, 1728, 1624, 1486, 1407, 1207 cm -1;
    [0188]
    [0189] H-NMR (500 MHz, D 2 O) 5 7.33 (d, J = 8.8 Hz, 1H), 7.11 (d, J = 2.0 Hz, 1H), 6.93 (dd, J = 8.8, 2.0 Hz, 1H), 6.67 (s, 1H), 4.55 (br s, 2H), 4.26 (t, J = 4.8 Hz, 2H), 3 , 93 (br s, 2H), 3.65 (s, 3H), 3.48 (d, J = 12.2 Hz, 2H), 3.42 (t, J = 4.8 Hz, 2H), 3.05 (m, 1H), 2.91 (t, J = 12.2 Hz, 2H), 2.84 (s, 3H), 1.83-1.80 (m, 2H), 1.69. -1.62 (m, 3H), 1.59-1.35 (m, 1H);
    [0190]
    [0191] 13 C-NMR (126 MHz, D 2 O) 5 164.5, 151.9, 133.8, 128.2, 126.5, 113.7, 111.4, 103.6, 80.2, 71, 3, 61.9, 55.4, 53.2 (2 C), 49.4, 44.2, 39.1.29.7, 22.4 (2 C), 20.8 (1 C).
    [0192] Analysis C 21 H 29 N 3 O 2 XHCO 2 CO 2 H 3/2 H 2 O:
    [0193] Calculated C, 54.94; H, 6.64; N, 7.69
    [0194] Found: C, 55.14; H, 6.39; N, 7.27
    [0195]
    [0196] Example 4: Synthesis of W-Methyl-W - ((1-methyl-5- (3- (piperidm-1-N) propoxy) -1H-indol-2-yl) methyl) prop-2-in-1 - amine
    [0197]
    [0198] Following the general synthesis of the compounds of formula (I), from 1-methyl-2 - ((methyl (prop-2-yn-1-yl) amino) methyl) -1H-indol-5-ol (Crosses , MA, Elorriaga, C., Fernández-Alvarez, E. Acetylenic and allenic derivatives of 2- (5-benzyloxyindolyl) and 2- (5-hydroxyindolyl) methylamines: synthesis and in vitro evaluation as monoamine oxidase inhibitors, Eur. J. Med. Chem. 1991 , 26, 33-41) (390 mg, 1.71 mmol) and 1- (3-chloropropyl) piperidine hydrochloride (338 mg, 1.71 mmol) in dry DMF (4 mL), reaction with NaH (205 mg, 5.13 mmol, 60% dispersion in mineral oil), after column chromatography (DCM / MeOH 1-5%), the title compound (400 mg, 76%) was obtained as a solid. : melting point 155-8 ° C
    [0199] Rf: 0.32 (DCM / 10% MeOH);
    [0200]
    [0201] IR (KBr) v 3435, 3269, 2942, 2505, 1620, 1489, 1207 cm -1;
    [0202]
    [0203] 1 H-NMR (500 MHz, CDC b) 57.17 (d, J = 8.8 Hz, 1H), 6.98 (d, J = 2.5 Hz, 1H), 6.78 (dd, J = 8 , 8, 2.5 Hz, 1H), 6.35 (s, 1H), 4.08 (t, J = 5.3 Hz, 2H), 3.74 (s, 3H), 3.71 (s) , 2H), 3.56 (d, J = 11.2 Hz, 2H), 3.33 (s, 2H), 3.18 (m, 2H), 2.66-2.62 (m, 2H) , 2.47, 2.45 (m, 2H), 2.32 (s, 3H), 2.41-2.27 (m, 2H), 1.92-1.83 (m, 3H), 1, 42-1.39 (m, 1H);
    [0204] 13 C-NMR (126 MHz, CCCb) 5 165.0, 152.4, 133.6, 127.5, 111.6, 109.8, 103.5, 102.4, 65.7, 55.5, 53.4 (2C), 51.5, 44.6, 41.4, 31.5, 30.0, 24.0, 22.5, 22.1 (2 C), 14.1;
    [0205]
    [0206] MS (ESI) m / z: 354.2 (M + 1) +.
    [0207]
    [0208] HMRS calculated for C 22 H 31 N 3 O: 354.2540; found 354.2536.
    [0209]
    [0210] The obtained compound was transformed into its bis-hydrochloride: melting point 220-1 ° C;
    [0211] 1 H-NMR (500 MHz, D 2 O) 57.33 (d, J = 9.3 Hz, 1H), 7.12 (d, J = 2.4 Hz, 1H), 6.92 (dd, J = 8.8, 2.4 Hz, 1H), 6.66 (s, 1H), 4.52 (s, 2H), 4.06 (t, J = 5.6 Hz, 2H), 3.92 (d, J = 1.9 Hz, 2H), 3.64 (s, 3H), 3.43 (d, J = 12.3 Hz, 2H), 3.16 (t, J = 7.0 Hz , 2H), 3.06 (t, J = 1.9 Hz, 1H), 2.83 (s, 3H), 2.83-2.77 (m, 2H), 2.10-2.07 ( m, 2H), 1.83-1.80 (m, 2H), 1.77-1.55 (m, 1H), 1.60-1.57 (m, 2H), 1.39-1, 36 (m, 1H);
    [0212]
    [0213] 13 C-NMR (126 MHz, CCCI 3 ) 5 152.3, 133.9, 128.4, 126.8, 113.9, 111.5, 105.9, 104.1, 80.3, 71.5 , 66.1, 54.5, 53.2 (2 C), 49.6, 44.3, 39.2, 29.8, 23.4, 22.7 (2 C), 21.0,
    [0214]
    [0215] Analysis. C 22 H 31 N 3 O 2 HCI:
    [0216] Calculated: C, 74.75; H, 8.84; N, 11.89
    [0217] Found: C, 74.81; H, 8.62; N, 11.61
    [0218]
    [0219] Example 5: Synthesis of W-Methyl-W - ((1-metN-5 - ((5- (piperidm-1-yl) pentyl) oxy) -1H-indol-2-yl) methyl) prop-2-in -1-amine
    [0220]
    [0221] Following the general synthesis of the compounds of formula (I), from 1-methyl-2 - ((methyl (prop-2-yn-1-yl) amino) methyl) -1H-indol-5-ol (Crosses , MA, Elorriaga, C., Fernández-Alvarez, E. Acetylenic and allenic derivatives of 2- (5-benzyloxyindolyl) and 2- (5-hydroxyindolyl) methylamines: synthesis and in vitro evaluation as monoamine oxidase inhibitors, Eur. J. Med. Chem. 1991 , 26, 33-41) (150 mg, 0.66 mmol) and 1- (5-chloropentyl) piperidine hydrochloride (150 mg, 0.66 mmol) in dry DMF (10 mL), reaction with NaH (48 mg, 1.97 mmol, 60% dispersion in mineral oil) after column chromatography (hexane / EtOAc, 10-50%), the title compound (175 mg, 70%) was obtained as a solid White.
    [0222]
    [0223] Rf = 0.33 (hexane / EtOAc 70%)
    [0224]
    [0225] 1 H-NMR (500 MHz, CDC b) 57.16 (d, J = 8.8 Hz, 1H), 6.99 (J = 2.7 1H), 6.81 (dd, J = 8.9, 2 , 7 Hz, 1H), 6.33 (s, 1H), 3.97 (t, J = 5.8 Hz, 2H), 3.71 (s, 3H), 3.65 (s, 2H), 3.59-3.37 (m, 2H), 3.29 (d, J = 2.1 Hz, 2H), 2.91 (t, J = 8.4 Hz, 2H), 2.68-2 , 53 (m, 2H), 2.32 (s, 3H), 2.28 (t, J = 2.1 Hz, 1H), 2.30-2.10 (m, 2H), 2.02- 1.89 (m, 4H), 1.82-1.79 (m, 4H), 1.60-1.47 (m, 2H);
    [0226] 13 C-NMR (126 MHz, CDCl 3 ) 5 153.0, 137.1, 133.4, 127.5, 111.8, 109.6, 103.4, 102.0, 78.3, 73.5 , 68.1, 57.3, 53.1 (2 C), 51.7, 44.7, 41.5, 29.9, 28.8, 23.7, 23.2, 22.5 (2 C), 22.1;
    [0227]
    [0228] MS (ESI) m / z: 382.3 (M + 1) +.
    [0229]
    [0230] The compound obtained was transformed into its bis-oxalate: melting point 123-6 ° C;
    [0231] IR (KBr) v 3431, 3263, 2946, 2868, 2680, 2541, 2124, 1724, 1623, 1537, 1486, 1473, 1405, 1280, 1207 cm-1
    [0232]
    [0233] Analysis C 24 H 35 N 3 O 2 XHCO 2 CO 2 H 2 H 2 O:
    [0234] Calculated C, 56.27; H, 7.25; N, 7.03
    [0235] Found: C, 56.66; H, 6.77; N, 7.04
    [0236]
    [0237] Farm studies
    [0238]
    [0239] Based on the methods described below, the values indicated in Table 1 have been obtained. Table 1 also shows the values of the following tests obtained for the ASS234, clorgiline, deprenyl and donepezil compounds for comparative purposes. .
    [0240]
    [0241] Inhibition studies of acetylcholinesterase and butyrylchinestarase
    [0242]
    [0243] The inhibitory activity of the enzyme acetylcholinesterase (AChE) was evaluated by the Ellman method ( Biochem Pharmacol , 1961, 7, 88) using the electric eel AChE model ( Electrophorus electricus) and acetylthiocholine iodide (0.35 mM) as a substrate. The reaction took place in a final volume of 3 ml of 0.1M phosphate buffer solution, pH 8.0, containing 0.035 units of AChE and a 0.35 mM solution of 5,5'-dithiobis (2-nitrobenzoic acid) was used. ) (DTNB) to produce the 5-thio-2-nitrobenzoic acid anion. Inhibition curves were performed in triplicate by incubating with at least nine concentrations of inhibitor for 10 min. A triplicate sample without inhibitor was always present to know 100% of AChE activity. After this time, the iodide substrate acetylthiocholine at 0.35 mM was added from a 10 mM stock solution. The color loss was observed at 412 nm in a reader spectrophotometric of 96-well plates. Determinations of the BuChE inhibitory activity, extracted from horse serum, were carried out in a similar manner, using 0.05 units / ml of BuChE, 0.35 mM of 5,5'-dithiobis-2-nitrobenzoic acid (DTNB ) and 0.5 mM butyrylthiocholine iodide from a 10 mM stock solution, in a final volume of 3 ml. A triplicate sample without inhibitor was always present to know 100% activity of the BuChE enzyme. The data from the concentration-inhibition experiments of the inhibitors were calculated by non-linear regression analysis, using the Origin package, which gives estimates of the IC 50 (concentration of the drug that produces 50% inhibition of the activity of the enzyme). The results expressed as mean ± SEM of at least four experiments carried out in triplicate. DTNB, acetylthiocholine iodide, butyrylthiocholine iodide.
    [0244]
    [0245] Inhibition studies of monoaminoxidases ( MAO)
    [0246]
    [0247] The inhibitory activity of the monoaminoxidases A and B was evaluated by the radiometric method of Fowler and Tipton ( Biochem Pharmacol 1981, 30, 3329) using a purification of mitochondria from rat liver as a source of the enzymes. The MAO-B inhibitory activity was performed against 25 μl of [14 C] -phenylethylamine (PEA) 20 μM of activity 2.5 μCi / mmol. The MAO-A inhibitory activity was performed against 25 μl of [14 C] - (5-hydroxytryptamine) (5-HT) 100 μM activity 0.5 mCi / mmol. Inhibition curves were performed in triplicate by incubating with at least ten concentrations of inhibitor for 30 minutes. A triplicate sample without inhibitor was always present in order to know 100 % MAO activity. The reaction took place with the addition of the substrate in a final volume of 225 μl of 50 mM phosphate buffer, pH 7.4 containing 20 μl of rat liver mitochondria at a concentration of 5 mg / ml. The reaction was always carried out under agitation, at 37 ° C and for 4 min in the case of MAO-B and 20 min in the case of MAO-A. The assay ended with the addition of 100 jl of 2M citric acid. The aldehydes produced were obtained after adding 4 ml of a solution of toluene: ethyl acetate (1: 1, v / v) containing 0.6% (w / v) 2,5-diphenyloxazole (PPO) and stirring the vials for 1 min leaving them at -80 ° C for 20 min. In this way, the freezing of the aqueous phase occurred, where the non-metabolized substrate was found, and the phase was decanted organic where the aldehyde produced is found. The radioactivity of the organic phase was read on a Tri-Carb 2810TR scintillation counter with a count time of 1 min per vial. From the data of disintegrations per minute (dpm) obtained, the specific activity of the enzyme (pmol / min-mg protein) was calculated with the following equation:
    [0248]
    [0249] dpm • (100 / X) • Y • (1 / t reaction in min) • (1 / ^ l prot) • (1000 / P) = pmol / min-mg prot
    [0250] where:
    [0251] X is the extraction ratio of the aldehyde in the organic phase {Fowler, 198057 / id} and this is 74.4 % for the serotonin aldehyde and 92.5% for the phenylethylamine aldehyde. Y is the conversion factor from dpm to pmol, which depends on the activity of the substrate, and is 0.9 for serotonin and 0.18 for phenylethylamine. Finally, P is the concentration of protein used expressed in mg / ml.
    [0252]
    [0253] The data were calculated by non-linear regression analysis, sigmoidal dose-response, using the GraphPad Prism 3.0 program, from which the IC 50 estimates for each of the inhibitors were obtained. The results have been expressed as mean ± SEM of at least three experiments carried out in triplicate.
    [0254]
    [0255] Interaction with hH3R / hH4R
    [0256]
    [0257] hH3R
    [0258]
    [0259] HEK-293 cells, which express hH3R, were washed and stored in a PBS buffer; then, they were centrifuged (3,000 g, 10 min, 4 ° C) and homogenized with an Ultraturrax in a cold buffer (12.5 mM MgCh, 100 mM NaCl and 75 mM Tris / HCl, pH 7.4). The homogenate of the membrane cells was centrifuged (20,000g, 20 min, 4 ° C), the pellet obtained was re-suspended in the buffer and stored at -80 ° C until use.
    [0260]
    [0261] Before beginning the experiments, the membrane cells were thawed, homogenized at 4 ° C and stored in a cold buffer. The membranes (20 Mg / well in a final volume of 0.2 ml of buffer) were incubated with [3 H] N-alphamethylhistamine (2 nM, 78.3 Ci / mmol) and different concentrations of the ligands test. The tests were carried out in triplicate to appropriate concentrations between 0.01 nM and 100 m M of the test compound. Incubations were performed for 90 min at room temperature. Non-specific binding was determined in the presence of 10 m M pitolisant. The bound radioligand was separated from the free radioligand by filtration using GF / B filters pre-treated with 0.3% polyethyleneimine (m / v), using an Inotech cell preservative. The radioactivity was determined by liquid scintillation counting. The data was analyzed with the GraphPad Prism 6 software using the non-linear regression method.
    [0262]
    [0263] hH4R
    [0264]
    [0265] Sf-9 cells were co-infected with baculovirus containing hH4R, G-protein Gai 2 and GP 1 Y 2 for 48 h. The infected cells were centrifuged (1,000 g, 10 min, 4 ° C) and washed with buffer (12.5 mM MgCh, 1 mM EDTA and 75 mM Tris / HCl, pH 7.4). For lysis of the cells, they were re-suspended in a cold lysis buffer (10 mM Tris / HCl (pH 7.4), 1 mM EDTA, 0.2 mM phenylmethylsulfonyl fluoride, 10 mg / ml benzamidine and leupeptin 10 Mg / ml) and homogenized in a mortar. The homogenate of the membrane cells was centrifuged (18,000g, 20 min, 4 ° C), the obtained pellet was re-suspended in the buffer and stored at -80 ° C until use.
    [0266]
    [0267] Before starting the experiments the membrane cells were thawed, homogenized at 4 ° C and stored in a cold buffer. The membranes (40 Mg / well in a final volume of 0.2 ml of buffer) were incubated with [3 H] histamine (10 nM, 10.6 Ci / mmol) and different concentrations of the compounds to be tested. The assays were carried out in triplicate at the appropriate concentrations between 0.01 nM and 100 mM of the test compound. Incubations were performed for 60 min at room temperature. The non-specific binding was determined in the presence of 100 mM of JNJ7777120. The following steps were performed as described above for hH3R.
    [0268]
    [0269] Analysis of antioxidant capacity
    [0270]
    [0271] The antioxidant activity of the compounds to be tested was determined by the "Oxygen Radical Absorbance Capacity-Fluorescein" method (ORAC-FL), using 2,2'-azobis (amidinopropane) dihydrochloride (AAPH) as a source of peroxyl radicals at 37 ° C. The reaction mixture was brought to 37 ° C in a 75 mM phosphate buffer (pH 7.4). First, a solution of the antioxidant (20 μl) and fluorescein (FL, 120 μl, final concentration 70 nM) were incubated in a 96-well microplate (Nunc) for 15 min at 37 ° C in a Varioskan plate reader Flash with injectors (Thermo Scientific). Next, the AAPH solution (60 μl, final concentration 12 mM) was added rapidly using the injector and the fluorescence was measured every minute for 60 min at Aex = 485 nm and Aem = 535 nm. The white mixture was composed of 120 μl of FL, 60 μl of AAPH and 20 μl of phosphate buffer (pH = 7.4). Trolox was used as a standard at concentrations between 1-8 [mu] M, and the compounds to be tested were measured at different concentrations between 0.1-1 [mu] M. All assays were done in triplicate and at least three different assays per sample. The measurement of fluorescence was normalized with a blank (without antioxidant and the area under the curve (AUC) showing the decrease in fluorescence was calculated by the formula: AUC = 1 sum (fi / f0), where f0 is the initial fluorescence at 0 min and fi is the fluorescence at time I. The net AUC for each sample was calculated as follows:
    [0272]
    [0273] AUC net = AUC antioxidant - AUCwhite.
    [0274]
    [0275] The equations of the regression were extrapolated representing the net AUC versus the concentration of the antioxidant. The ORAC values correspond to the ratio of the slopes of the last curve and that of Trolox in the same experiment. The final ORAC values are expressed as Trolox equivalents and the data as means ± SD.
    [0276]
    [0277] As observed in the above Table, the compounds of the invention of Examples 1 to 5 demonstrate having interesting pharmacological properties for their selection and identification as multipotent molecules for the potential treatment of neurological diseases, for example.
    [0278]
    [0279] Thus, in particular with reference to the values in Table 1 it can be deduced that, among all the compounds prepared and evaluated, the compound W-methyl-W - ((1-methyl-5- (3- (piperidin-1-yl) ) propoxy) -1H-indol-2-yl) methyl) prop-2-in-1-amine (Ex. 4) behaves as a potent antioxidant agent, similar in potency to ferulic acid, a potent inhibitor of enzymes cholinesterases in the micromolar range (IC 50 <2 ^ M), as an irreversible, potent and selective inhibitor of MAO B (IC 50 = 78 nM, selectivity 0.5), more potent than the reference product ASS234 to inhibit MAO B, but less potent to inhibit MAO A. Finally, the compound W-methyl-W - ((1-methyl-5- (3- (piperidin-1-yl) propoxy) -1H-indol-2-yl) methyl) prop -2-in-1 -amine has a strong affinity for the H3R receptor (Ki = 10.8 nM) and selective against H4R.
    [0280]
    [0281] In sum, all the investigated pharmacological properties of the compound N-methyl-N - ((1-methyl-5- (3- (piperidin-1-yl) propoxy) -1H-indol-2-yl) methyl) prop-2 -in-1 -amine and the rest of the compounds of the invention are clearly superior to those found for the ASS234 molecule, which allows us to conclude that this compound is a very promising molecule to initiate in vivo studies for its use in the treatment of neurodegenerative and central nervous system diseases.
    权利要求:
    Claims (14)
    [1]
    1. Compound of formula (I)

    [2]
    2. Compound of formula (I) according to claim 1, wherein R3 is H.
    [3]
    3. Compound of formula (I) according to any of claims 1 or 2, wherein R1 and R2 are, independently of each other, a linear or branched, substituted or unsubstituted (C 1 -C 4 ) alkyl group.
    [4]
    4. Compound of formula (I) according to any of claims 1 to 3 wherein X is CH 2 .
    [5]
    5. Compound of formula (I) according to claim 1, wherein R3 is H, X is CH 2 and R1 and R2 are both a methyl group.
    [6]
    6. Compound of formula (I) according to any of the preceding claims, which is selected from among
    • W-Methyl-N - ((1-methyl-5- (2- (pyrrolidin-1-yl) ethoxy) -1 H -indol-2-yl) methyl) prop-2-yn-1 -amine
    • W-Methyl-W - ((1-methyl-5- (3- (pyrrolidin-1-yl) propoxy) -1 H -indol-2-yl) methyl) prop-2-yn-1-amine
    • W-Methyl-W - ((1-methyl-5- (2- (piperidin-1-yl) ethoxy) -1 H -indol-2-yl) methyl) prop-2-in-1 -amine
    • W-Methyl-W - ((1-methyl-5- (3- (piperidin-1-yl) propoxy) -1 H -indol-2-yl) methyl) prop-2-yn-1-amine
    • W-Methyl-W - ((1-methyl-5 - ((5- (piperidin-1 -yl) pentyl) oxy) -1 H -indol-2-yl) methyl) prop-2-in-1- amine.
    [7]
    7. Compound of formula (I) according to claim 6, which is N-Methyl-N - ((1-methyl-5- (3- (piperidin-1-yl) propoxy) -1H-indol-2-yl) methyl) prop-2-in-1 -amine.
    [8]
    8. Process for obtaining the compounds of formula (I) according to claim 1, comprising the reaction of a compound of formula (III)

    [9]
    9. A pharmaceutical composition comprising at least one compound of formula (I) according to any of claims 1 to 7 and at least one pharmaceutically acceptable adjuvant, excipient and / or vehicle and / or, if appropriate, one or more pharmacologically active compounds. additional
    [10]
    10. Pharmaceutical composition according to claim 9, characterized in that it is in a solid form or in aqueous suspension, in a diluent pharmaceutically acceptable, for oral, topical, rectal or parenteral administration.
    [11]
    11. Pharmaceutical composition according to claims 9 to 10, characterized in that it includes the compounds of formula (I) in the form of mono- or bisoxalate or hydrochloride salts.
    [12]
    12. Use of a compound of the formula of (I) according to claim 1 for the manufacture of a medicament for the treatment of a neurodegenerative or central nervous system disease.
    [13]
    13. Use of a compound of the formula of (I) according to claim 12, characterized in that the neurodegenerative or central nervous system disease is selected from senile dementia, cerebrovascular dementia, mild cognitive deficit, attention deficit disorders, neurodegenerative diseases associated with aggregations. of aberrant proteins such as Parkinson's disease or Alzheimer's disease, amyotrophic lateral sclerosis, prion diseases such as Creutzfeldt-Jakob disease or Gerstmann-Straussler-Scheinker disease, stroke, amyotrophic lateral sclerosis, narcolepsy, and sleep disturbances and disorders .
    [14]
    14. Use of a compound of the formula of (I) according to claim 13, characterized in that the neurodegenerative or central nervous system disease is Parkinson's disease or Alzheimer's disease.
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    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    WO2011113988A1|2010-03-18|2011-09-22|Consejo Superior De Investigaciones Científicas |New derivatives of propargylamine having neuroprotective capacity for the treatment of alzheimer's and parkinson's diseases|
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