WO2012113967A1

WO2012113967A1 - Gsk-3 inhibitors that can be used in neurodegenerative and inflammatory diseases, cancer, diabetes and in regenerative processes

Info

Publication number: WO2012113967A1
Application number: PCT/ES2012/070119
Authority: WO
Inventors: Ana Martinez Gil; Carmen Gil Ayuso-Gontan; Mª del Valle PALOMO RUIZ; Concepción PEREZ MARTIN; Daniel Ignacio Perez Fernandez; Francisco Javier Luque Garriga
Original assignee: Consejo Superior De Investigaciones Cientificas (Csic); Universidad De Barcelona
Priority date: 2011-02-25
Filing date: 2012-02-27
Publication date: 2012-08-30
Also published as: ES2387359A1; ES2387359B1

Abstract

The present invention relates to a compound of formula (I) derived from maleimide and that has the ability to reversibly or irreversibly inhibit the GSK-3 enzyme in the micro- and nanomolar ranges. The present invention likewise relates to the use of these compounds for producing a drug for treating and/or preventing diseases in which GSK-3 is implicated, such as neurodegenerative diseases, inflammatory diseases, cancer or diabetes. These compounds can also be used to promote regenerative processes in various tissues.

Description

GSK-3 inhibitors useful in neurodegenerative, inflammatory diseases, cancer, diabetes and in reqenerative processes.

The present invention relates to a series of compounds that are derived from maleimides and that are capable of inhibiting the GSK-3 enzyme in micro and nanomolar ranges reversibly or irreversibly. These compounds, therefore, are useful for the manufacture of a medicament for the treatment and / or prevention of diseases in which GSK-3 is involved, such as neurodegenerative diseases, inflammatory diseases, cancer, diabetes, as well as promoting various processes. regenerative

STATE OF THE PREVIOUS TECHNIQUE

Glycogen synthase kinase 3, GSK-3, is an enzyme in the kinase family that catalyzes phosphorylation of serine or threonine residues in various substrates. It was originally discovered for its role in glycogen biosynthesis, to which it owes its name [Rylatt, DB, Aitken, A., Bilham, T., Condom, GD, Embi, N., Cohen, P. "Glycogen Synthase Kinase 3 from rabbit skeletal muscle. Separation from cyclic-AMP-dependent protein kinase and phosphorylase kinase ". Eur J Biochem. 1980 107, 519-527.]. The enzyme is involved in the regulation of several cell signaling pathways, including Wnt pathways, the cell division cycle, DNA damage response, cell death and survival, and neuronal differentiation among others [ Van Waue, J., Haefner, B. "Glycogen Synthase Kinase-3 as drug target: from wallflower to center of attention". Drug News Perspect. 2003 16, 557-565]. Recent studies show that overexpression of GSK-3 is sufficient to induce neuronal death [Hetmán, M., Cavanaugh, JE, Kimelman, D., Xia, Z. "Role of glycogen synthase kinase-3beta in neuronal apoptosis induced by trophic withdrawal ". J Neuroso. 2000 20, 2567-2574], relating to various pathologies such as bipolar disorders, neurodegenerative diseases, especially Alzheimer's disease, type II diabetes and chronic inflammatory diseases. Recently, some groups have suggested that GSK-3 plays an important role in cell proliferation and differentiation signals because of its essential role in the signaling pathways of RTK, Wnt and Shh. Mesemchymal stem cells (MSC) have the ability to differentiate into several cell types, including osteoblasts. Activation of Wnt signaling by inhibition of GSK-3 causes the differentiation of MSCs into osteoblasts and the consequent increase in bone mass [Gambardella A, Nagaraju CK, O'Shea PJ, Mohanty ST, Kottam L, Pilling J, Sullivan M, Djerbi M, Koopmann W, Croucher Pl, Bellantuono I "Glycogen synthase kinase-3a / p inhibition promotes in vivo amplification of endogenous mesenchymal progenitors with osteogenic and adipogenic potential and their differentiation to the osteogenic lineage". J Bone Miner Res. 2010 Oct 1 1]. Likewise, it has also been shown that inhibition of GSK-3 promotes cardiomyocyte proliferation in the heart of adults, so inhibiting GSK3 could be a strategy to promote cardiac regeneration in pathological states [Woulfe KC, Gao E, Lal H , Harris D, Fan Q, Vagnozzi R, DeCaul M, Shang X, Patel S, Woodgett JR, Forcé T, Zhou J. "Glycogen synthase kinase-3beta regulates post-myocardial infarction remodeling and stress-induced cardiomyocyte proliferation in vivo" Circ Res. 2010 May 28; 1 06 (10): 1 635-45].

In recent years, numerous GSK-3 inhibitors have been synthesized, resulting promising molecules for the treatment of various diseases, such as diabetes, cancer and neurodegenerative diseases [Martínez, A. "Preclinical efficacy on GSK-3 inhibitors: towards a future generation of powerful drugs. " Med. Res. Rev. 2008 28, 773-796]. In addition, the inhibition of kinases that inhibit tau protein could be beneficial for neurodegenerative diseases [Martínez, A., Castro, A. "Inhibition of tau phosphorylation: A new therapeutical strategy for the treatment of Alzheimer's disease and other neurodegenerative disorders" Expert Opinion . Ther. Pal 2000 10, 1 51 9-1 527]. Specifically in these types of diseases, these inhibitors are demonstrating their effectiveness, and today there is a GSK-3 inhibitor that is in clinical phases for the treatment of Alzheimer's disease, and for progressive supranuclear paralysis. DESCRIPTION OF THE INVENTION

The present invention presents a family of four compounds, and their mode of production, which possess the ability to inhibit the GSK-3 enzyme in micro and nanomolar order. In addition, some of these compounds, according to a model adapted to the laboratory, are capable of crossing the blood-brain barrier, a key feature for compounds that can serve as drugs for diseases of the central nervous system. In a first aspect, the present invention relates to a compound of formula (I):

(I)

where R ¹ is selected from H or C C ₀ alkyl and R ² is selected from dC-io O alkyl Crdo alkenyl, optionally substituted by halogen, or its pharmaceutically acceptable salts, solvates or isomers.

The term "alkyl" refers, in the present invention, to radicals of hydrocarbon chains, linear or branched, having 1 to 10 carbon atoms, preferably 1 to 4, and which are attached to the rest of the molecule by a single bond, for example, methyl, ethyl, n-propyl, / -propyl, n-butyl, tere-butyl, sec-butyl, n-pentyl, n-hexyl, etc. The alkyl groups may be optionally substituted by one or more halogen atoms, that is, fluorine, chlorine, bromine and iodine, preferably bromine. The term "alkenyl" refers to radicals of linear or branched hydrocarbon chains, having 1 to 10 carbon atoms, preferably from 1 to 4 and containing one or more double carbon-carbon bonds, for example, vinyl, 1-propenyl, allyl, isoprenyl, 2-butenyl, 1, 3- butadienyl, etc. Alkenyl radicals may be optionally substituted by one or more halogen atoms.

In a preferred embodiment, R ¹ is H.

In another preferred embodiment, R ¹ is CC ₄ alkyl and more preferably methyl. In a more preferred embodiment, R ² is CrC ₄ alkyl and more preferably methyl.

In another more preferred embodiment, R ² is -CH ₂ Br. In a preferred embodiment, the compound of formula (I) is selected from the list comprising:

^■ 3-acetyl-4- (1 H-indole-3-yl) -1 H-pyrrole-2,5-dione

^■ 3-Acetyl-4- (1-methyl-1 H-indole-3-yl) -1 H-pyrrole-2,5-dione

^■ 3- (2-Bromoacetyl) -4- (1 H-indole-3-yl) -1 H-pyrrole-2,5-dione

■ 3- (2-Bromoacetyl) -4- (1 H-indole-3-yl) -1 H-pyrrole-2,5-dione

or of its pharmaceutically acceptable salts, solvates or isomers.

The compounds of the present invention represented by 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. The individual isomers, enantiomers or diastereoisomers and mixtures thereof fall within the scope of the present invention, that is, the term isomer also refers to any mixture of isomers, such as diastereomers, racemic, etc., even their optically isomers. assets or mixtures in different proportions thereof. The individual enantiomers or diastereoisomers, as well as mixtures thereof, can be separated by conventional techniques. The compounds of the invention may be in crystalline form as free compounds or as solvates. In this sense, the term "solvate", as used herein, includes both pharmaceutically acceptable solvates, that is, solvates of the compound of formula (I) that can be used in the manufacture of a medicament, as pharmaceutically acceptable solvates, which may be useful in the preparation of pharmaceutically acceptable solvates or salts. The nature of the pharmaceutically acceptable solvate is not critical as long as it is pharmaceutically acceptable. In a particular embodiment, the solvate is a hydrate. Solvates can be obtained by conventional solvation methods known to those skilled in the art.

Another aspect of the invention relates to the use of a compound of formula (I) for the manufacture of a medicament.

Another aspect of the invention relates to the use of a compound of formula (I) for the manufacture of a medicament for the treatment and / or prevention of a disease that is selected from neurodegenerative diseases, inflammatory diseases, cancer or diabetes.

Preferably, the neurodegenerative disease is selected from Alzheimer's disease, Parkinson 's disease, amyotrophic lateral sclerosis, cerebral ischemia, post-encephalitic Parkinsonism, dystonia, Tourette ^'s, pathologies limb movements newspapers, restless leg syndrome, deficit disorders of attention with hyperactivity, Hungtinton's disease, progressive supranuclear paralysis, Pick's disease, frontotemporal dementia, neuromuscular diseases.

Preferably, the inflammatory disease is selected from Crohn's disease, ulcerative colitis, arthritis, atherosclerosis, vasculitis, multiple sclerosis. Preferably, the cancer is selected from glioblastoma, leukemia, lymphoma, lung, breast, prostate or colon cancer and in general from any cancerous or metastatic process in which GSK-3 is involved. Preferably, diabetes is selected as non-dependent insulin type II diabetes.

Another aspect of the invention is the use of a compound of formula (I) for the manufacture of a medicament to promote regenerative processes. Preferably, the regenerative processes are selected from those that promote the differentiation of the stem cells of the nervous system, the hematopoietic system, the bone system, the myocardium.

For their application in therapy, the compounds of formula (I), their salts, solvates or isomers, will preferably be found in a pharmaceutically acceptable or substantially pure form, that is, having a pharmaceutically acceptable level of purity excluding pharmaceutical additives. normal 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), or of its salts, solvates or isomers. The compounds of the present invention of formula (I) can be obtained according to the procedure described in the document [Faul, M.M., Winneroski, L.L., Brussee, J., Krumrich, C.A. "A new one step synthesis of maleimides by condensation of glyoxylate esters with acetamides". Tetrahedron Letters 1999 40, 1 109-1 1 12].

In another aspect, the present invention also relates to pharmaceutical compositions comprising at least one compound of the invention, or an isomer, a pharmaceutically acceptable salt, a derivative or a prodrug. thereof, together with a pharmaceutically acceptable carrier or carrier, an excipient or a vehicle, for administration to a patient.

In a preferred embodiment, the pharmaceutical composition further comprises another active ingredient.

The pharmaceutically acceptable adjuvants and vehicles that can be used in said compositions are the adjuvants and vehicles known to those skilled in the art and commonly used in the elaboration of therapeutic compositions.

Another aspect of the invention relates to a compound of formula (I) for use as a medicament and particularly as a medicament for treating and / or preventing neurodegenerative diseases, inflammatory diseases, cancer or diabetes or for promoting regenerative processes.

Another aspect of the invention is a method of treating a neurodegenerative disease, an inflammatory disease, cancer or diabetes, as well as a method of promoting cell regeneration, which comprises administering to a patient a therapeutically effective amount of a compound of formula (I) or of a pharmaceutical composition comprising it,

In the sense used in this description, the term "therapeutically effective amount" refers to the amount of the agent or compound capable of developing the therapeutic action determined by its pharmacological properties, calculated to produce the desired effect and, in general, will be determined, among other causes, due to the characteristics of the compounds, including the age, condition of the patient, the severity of the alteration or disorder, and the route and frequency of administration.

The compounds described in the present invention, their salts, prodrugs and / or solvates as well as the pharmaceutical compositions containing them can be used together with other drugs, or active ingredients, additional to provide a combination therapy. Said additional drugs may be part of the same pharmaceutical composition or, alternatively, they may be provided in the form of a separate composition for simultaneous or non-simultaneous administration to the pharmaceutical composition comprising a compound of formula (I), or a salt, prodrug or solvate thereof.

In another particular embodiment, said therapeutic composition is prepared in the form of a solid form or aqueous suspension, in a pharmaceutically acceptable diluent. The therapeutic composition provided by this invention may be administered by any appropriate route of administration, for which said composition will be formulated in the pharmaceutical form appropriate to the route of administration chosen. In a particular embodiment, the administration of the therapeutic composition provided by this invention is performed orally, topically, rectally or parenterally (including subcutaneously, intraperitoneally, intradermally, intramuscularly, intravenously, etc.).

In a preferred embodiment of the present invention, the pharmaceutical compositions are suitable for oral administration, in solid or liquid form. Possible forms for oral administration are tablets, capsules, syrups or solutions and may contain conventional excipients known in the pharmaceutical field, as aggregating agents (eg syrup, acacia, gelatin, sorbitol, tragacanth or polyvinyl pyrrolidone), fillers (eg lactose, sugar, corn starch, calcium phosphate, sorbitol or glycine), disintegrants (eg starch, polyvinyl pyrrolidone or microcrystalline cellulose) or a pharmaceutically acceptable surfactant such as sodium lauryl sulfate.

Compositions for oral administration can be prepared by conventional methods of Galenic Pharmacy, as mixing and dispersion. The tablets can be coated following methods known in the pharmaceutical industry. The pharmaceutical compositions can be adapted for parenteral administration, as sterile solutions, suspensions, or lyophilized products of the invention, using the appropriate dose. Suitable excipients, such as pH buffering agents or surfactants, can be used.

The aforementioned formulations can be prepared using conventional methods, such as those described in the Pharmacopoeias of different countries and in other reference texts. The administration of the compounds or compositions of the present invention can be performed by any suitable method, such as intravenous infusion and oral, intraperitoneal or intravenous routes. Oral administration is preferred for the convenience of patients and for the chronic nature of the diseases to be treated.

The amount administered of a compound of the present invention will depend on the relative efficacy of the compound chosen, the severity of the disease to be treated and the weight of the patient. However, the compounds of this invention will be administered one or more times a day, for example 1, 2, 3 or 4 times daily, with a total dose between 0.1 and 1000 mg / kg / day. It is important to keep in mind that it may be necessary to introduce variations in the dose, depending on the age and condition of the patient, as well as modifications in the route of administration. The compounds and compositions of the present invention can be used together with other medicaments in combination therapies. The other drugs may be part of the same composition or of a different composition, for administration at the same time or at different times. The use of the compounds of the invention is compatible with their use in protocols in which the compounds of the formula (I), or mixtures thereof are used by themselves or in combinations with other treatments or any medical procedure. Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and features of the invention will be derived partly from the description and partly from the practice of the invention. The following examples and drawings are provided by way of illustration, and are not intended to be limiting of the present invention. DESCRIPTION OF THE FIGURE

Figure 1. Shows the linear correlation between described and experimental permeability of 10 commercial compounds using the PAMPA-Blood-brain barrier methodology.

EXAMPLES

The invention will now be illustrated by tests carried out by the inventors, which show the specificity and effectiveness of the compounds of the invention.

Synthesis of 3,4-disubstituted maleimides

Comp. 1 R ¹ = H Comp. 3 R ¹ = H

Comp. 2 R ¹ = Me Comp. 4 R ¹ = Me

3-acetyl-4- (1 H-indol-3-yl) -1 H-pyrrolin-2,5-dione (Compound 1): On a mixture of acetoacetamide (1 equiv, 3.69 mmol, 372 mg) and ^f BuOK (1 M solution in THF) (3.5 equiv, 12.9 mmol, 12.9 mi) dissolved in THF (20 mi) at -60 ° -C methyl 3-indolglioxylate (1 equiv, 3.69 mmol, 750 mg) is added. The mixture is stirred until room temperature is reached and 1.5 ml of concentrated hydrochloric acid, 30 ml of water and 30 ml of dichloromethane are added. Then, the organic phase is separated and washed with a saturated solution of NaHC0 ₃ (2 x 30 ml). It is dried with MgSO ₄ and the solvent is evaporated under reduced pressure. The solid is purified by recrystallization from ethyl acetate / pentane to obtain an orange solid, 0.65 g, yield: 74%. ¹ H NMR (300 MHz, DMSO-cfe): δ 12.29 (sa, 1 H), 1 1, 17 (sa, 1 H), 8.24 (s, 1 H), 7.50 (d, J = 7.01 Hz, 1 H), 7.27 - 7.17 (m, 1 H), 7.13 (s, 2H), 2.50 - 2.46 (s, 3H). ¹³ C NMR (75 MHz, DMSO-cfe): δ 195.49, 171, 74, 139.80, 137.50, 135.53, 125.57, 124.94, 123.51, 122.67, 121 , 70, 1 13.36, 106.05, 31, 72, HPLC / MS: Purity> 99%, tr = 3.76 min, Sunfire C18 column, 3.5 μτη (50 X 4.6 mm) using as mobile phase acetonitrile (0.08% formic acid) and MiliQ water (0.1% formic acid) and a gradient of acetonitrile (10% to 100%) for 10 min with a flow of 0.25 ml / min (m / z 255,237.1 65). Mp = 225-226 ^Q C. Elemental Analysis (Ci ₄ Hi ₀ N ₂ O ₃ ) Calculated: C 66.14%; H 3.96%; N 1 1, 02%. Found: C 65.89%; H 4.12%; N 10.74%.

3-acetyl-4- (1-methyl-indole-3-yl) -1 H-pyrrolin-2,5-dione (Compound 2): On a mixture of acetoacetamide (1 equiv, 3.69 mmol, 372 mg) and ^f BuOK (1 M solution in THF) (3.5 equiv, 12.9 mmol, 12.9 ml) dissolved in THF (20 ml) at -60 ^Q C (1-methylindole) -3-glyoxylate is added methyl (1 equiv, 3.69 mmol, 3.69 mmol, 800 mg). The mixture is stirred until room temperature is reached and 1.5 ml of concentrated hydrochloric acid, 30 ml of water and 30 ml of dichloromethane are added. The organic phase is then separated and washed with a saturated solution of NaHC0 ₃ (2 x 30 ml). It is dried with MgSO ₄ and the solvent is evaporated under reduced pressure. The solid is purified by recrystallization from ethyl acetate / pentane to obtain a red solid, 0.55 g, yield: 56%. ¹ H NMR (300 MHz, DMSO-cfe): δ 1 1, 19 (s, 1 H), 8.27 (s, 1 H), 7.58 (d, J = 8.1 Hz, 1 H) , 7.35-7.06 (m, 3H), 3.92 (s, 3H), 2.51 (s, 3H). ¹³ C NMR (75 MHz, DMSO-cfe): δ 194.66, 170.97, 138.56, 138.14, 137.38, 125.28, 123.82, 122.84, 122.14, 121 , 31, 1 1 1, 08, 104.36, 33.30, 30.95. HPLC / MS: Purity = 95%, tr = 4.06 min, Sunfire C18 column, 3.5 μηπ (50 X 4.6 mm) using as mobile phase acetonitrile (0.08% formic acid) and MiliQ water (0.1% formic acid) and a gradient of acetonitrile (10% to 100%) for 10 min with a flow of 0.25 ml / min (m / z 269, 251, 180). Mp = 224-225 ° -C. Elemental Analysis (Ci ₅ H ₁₂ N ₂ 0 ₃ ) Calculated: C 66.60%; H 5.22%; N 10.36%. Found: C 66.89%; H 5.01%; N 10.37%.

3- (2-Bromoacetyl) -4- (1 H-indol-3-yl) -1 H-pyrrolin-2,5-dione (compound 3). Compound 1 (1 equiv, 2 mmol, 510 mg) is dissolved in acetic acid, and under reflux conditions bromine (1.05 equiv, 2.1 mmol, 0.1 1 mL) dissolved in acetic acid is added. The mixture is refluxed for 3 hours. After allowing the mixture to cool, water and ethyl acetate are added and the organic phase is extracted and washed with several portions of a saturated NaHC0 ₃ solution. The solvent is evaporated under reduced pressure and the solid is purified by recrystallization from MeOH / H ₂ 0 to obtain a black solid, 0.25 g, yield: 39%. ¹ H NMR (300 MHz, DMSO-cfe): δ 12.54 (s, 1 H), 1 1, 34 (s, 1 H), 8.39 (m, 1 H), 7.54 (d, J = 8.0 Hz, 1 H), 7.32-6.98 (m, 3H), 4.75 (s, 2H). ¹³ C NMR (126 MHz, acetone-efe): δ 186.0, 1 69.6, 1 69.4, 142.1, 135.9, 135.7, 124.7, 122.8, 121, 0 , 1 12.1, 106.4, 35.0, HPLC / MS: Purity = 95%, tr = 4.35 min, Sunfire C18 column, 3.5 μτη (50 X 4.6 mm) using as mobile phase acetonitrile (0.08% formic acid) and MiliQ water (0.1% formic acid) and a gradient of acetonitrile (10% to 100%) for 10 min with a flow of 0.25 ml / min (m / z 335 , 333, 253,182). Mp = 261-262 ° -C. Elemental Analysis (Ci ₄ H ₉ BrN ₂ 0 ₃ ) Calculated: C 50.47%; H 2.72%; N 8.41%; Br 23.99%. Found: C 50.68%; H 2.87%; N 8.61%; Br 23.62.

3- (2-Bromoacetyl) -4- (1-methyl-1 H-indol-3-yl) -1 H-pyrrolin-2,5-dione (compound

4). Compound 2 (1 equiv, 2.25 mmol, 603 mg) is dissolved in acetic acid, and under reflux conditions bromine (1.05 equiv, 2.36 mmol, 0.1 15 mL) dissolved in acid is added acetic. The mixture is refluxed for 3 hours. After allowing the mixture to cool, water and ethyl acetate are added and the organic phase is extracted and washed with several portions of a saturated NaHC0 ₃ solution. The solvent is evaporated under reduced pressure and the solid is purified by column chromatography on silica gel using as eluent CH ₂ CI ₂ / MeOH (80: 1) obtaining a purple solid, 0.12 g, yield: 15%. ¹ H NMR (300 MHz, CDCI ₃ ): δ 8.46 (s, 1 H), 7.38 (m, 4H), 5.31 (s, 1 H), 4.64 (s, 2H), 3.94 (s, 3 H). ¹³ C NMR (126 MHz, acetone-cfe): δ 185.80, 1 69.78, 1 69.55, 141, 80, 139.52, 137.66, 125.37, 122.94, 122.88 , 121, 34, 1 19.28, 1 10.46, 105.53, 35.02, 32.71, Mp = 252-253 ° -C. HPLC / MS: Purity = 99%, tr = 4.47 min, Sunfire C18 column, 3.5 μτη (50 X 4.6 mm) using acetonitrile (0.08% formic acid) and MiliQ water (0 as mobile phase) , 1% formic acid) and a gradient of acetonitrile (10% to 100%) for 10 min with a flow of 0.25 ml / min (m / z 347, 267, 196). Elemental Analysis (0Ι ₅ ΗΙ Ι ΒΓΝ ₂ Ο ₃ ) Calculated: C 51, 90%; H 3.19%; N 8.07%; Br 23.02%. Found: C 52.03%; H 3.08%; N 7.77%; Br 22.67%.

GSK-33 inhibition measure

Enzyme inhibition assays were performed using the kinasa-glo® luminometric method methodology. Recombinant human enzyme GSK3 (n ^Q catalog 14-306) was purchased from Upstate (Dundee, UK). The prefosphorylated polypeptide was synthesized by American Peptide Inc (Sunnyvale, CA). The luminescent kinase kit (n ^Q catalog V671 1) was obtained from Promega. ATP and other reagents were purchased in Sigma-Aldrich (St. Louis, MO)

The tests were performed in buffer using 96-well plates. 10μΙ of the compound to be tested (dissolved in dimethisulfoxide at a concentration of 1 mM, and in turn dissolved in buffer until the concentration necessary for the experiment) and 10μΙ (20ng) of the enzyme are added to each well followed by 20μΙ of buffer containing 25μΜ of the substrate and 1 μΜ of ATP. The final concentration of DMSO in the experiment did not exceed 1%. After incubation for half an hour at 30 ^Q C for enzymatic reaction with the reagent 40μΙ-Glo kinase. The luminescence is measured after ten minutes using a POLARstar Optima multimode reader. The activity is proportional to the difference between total and consumed ATP. Inhibition activities were calculated based on maximum activity, measured in the absence of inhibitor. Table 1. Inhibitory concentration of the compounds.

The four compounds evaluated show a very potent enzyme inhibition, with IC ₅ values or that vary from the low micromolar to low nanomolar range, irreversible inhibitors (3 and 4) being more potent than reversible ones (1 and 2). The step from reversible inhibitor (1 and 2) to irreversible (3 and 4) implies an increase in the inhibition of GSK-3 by an order of magnitude, respectively.

Central nervous system (CNS) penetration: Experiment One vitro) determination of permeability using parallel artificial membranes (PAMPA) of blood brain penetration.

The prediction of the permeability of various compounds on the central nervous system (CNS) was determined using the methodology of parallel artificial membranes (PAMPA) [Di, L., Kerns, EH, Fan, K., McConnell, OJ, Carter, GT "High throughput artificial membrane permeability assay for blood-brain barrier" Eur. J. Med. Chem., 2003, 38, 223-232]. Commercial compounds, pH 7.4 phosphate buffer (PBS), ethanol and dodecane were obtained from the Sigma, Acros organics, Merck, Aldrich and Fluka commercial houses. The porcine brain lipid (reference catalog 141 101) was purchased from Avanti Polar Lipids. Both the 96-well donor plate (Multiscreen® IP Sterile Píate membrane PDVF, pore size 0.45 μΜ, reference catalog MAIPS4510) and the 96-well acceptor plate (Multiscreen®, reference catalog MAMCS9610) were purchased from Millipore. In order to filter the samples, PDVF membrane filters (30 mm in diameter, pore size 0.45 μηπ) from the Symta commercial house were used. He The equipment used to perform ultraviolet absorbance measurements in 96-well plates was a Thermoscientific Multiskan spectrum.

Ten compounds were selected in order to validate the experiment. Different amounts of them were taken [(3-5 mg of Caffeine, Enoxacin, Hydrocortisone, Desipramine, Ofloxacin, Piroxicam, Testosterone), (12 mg of Promazine) and 25 mg of Verapamil and Atenolol] which were dissolved in EtOH ( 1000 μΙ_). 100 microliters of these solutions were taken and 1400 μΙ_ of EtOH and 3500 μΙ_ of phosphate buffer PBS (pH 7.4) buffer were added, in order to reach a final concentration of EtOH of 30% in the solution. The solutions were filtered. 180 μΙ of a solution of PBS / EtOH (70/30) was added to each well of the 96-well acceptor plate. The donor plate was impregnated with 4 μΙ_ of a solution of the porcine brain lipid dissolved in dodecane (20 mg mi ^“1 ). After 5 minutes, 180 μΙ_ of solution of each compound was added on this plate. To assess their penetration into the central nervous system, they were taken between 1-2 mg and dissolved in 1500 μΙ_ of EtOH and 3500 μΙ_ of phosphate buffer PBS (pH 7.4) buffer, filtered and added to the donor plate 96 Wells, the donor plate was then placed on the acceptor forming a kind of "sandwich" and left to incubate for 2h at 25 ° C. The compounds will pass from the donor plate through the porcine brain lipid to the acceptor plate. After 2 hours, the donor plate is carefully removed.The concentration and absorbance of both commercial compounds and synthesized derivatives that were evaluated in the acceptor and donor plates were determined using or n UV absorbance reader. Each sample was analyzed for 3 to 5 wavelengths, in 3 wells and in at least 2 independent experiments. The results are the average of the measurements [standard deviation (SD)] of the different experiments performed. 10 commercial compounds (previously discussed) whose penetration into the central nervous system is known, were used in each experiment in order to validate the method. Table 2. Permeability (Pe 10 ^"6 cm s ^{" 1} ) in the PAMPA-Blood-brain Barrier experiment for 10 commercial compounds (used for the validation of the experiment) and various derivatives synthesized with their corresponding prediction of central nervous system penetration (CNS) )).

Compound Pe (10 ⁶ cm s ¹ ) ³ Pe (10 ⁶ cm s ¹ ) "Prediction

Atenolol 0.8 0.2 ± 0.1

Caffeine 1, 3 0.9 ± 0.2

Desipramine 12 12.4 ± 1.0

Enoxacin 0.9 0.3 ± 0.2

Hydrocortisone 1, 9 0.6 ± 0.4

Ofloxacin 0.8 0.7 ± 0.7

Piroxicam 2.5 0.4 ± 0.4

Promazine 8.8 1 1, 5 ± 0.9

Testosterone 17 15.7 ± 1, 2

Verapamil 1 6 15.6 ± 1, 2

Comp. 1 1, 5 ± 0.5 SNC + / SNC -

Comp. 2 6.4 ± 1.5 SNC +

Comp. 3 1, 1 ± 0.3 SNC -

Comp. 4 3.9 ± 1.0 SNC +

PBS: EtOH (70:30) used as solvent. ^a Data estimated and presented in Article Eur. J. Med. Chem., 2003, 38, 223-232. ^b Data average ± standard deviation of at least 2 independent experiments.

These data suggest that compounds 2 and 4 may be more useful in pathologies mediated by GSK-3 that affect the central nervous system and that have the intact hematoencephalic barrier. While compounds 1 and 3, since the blood brain barrier does not pass in principle, they may be more useful in diseases mediated by GSK-3 peripheral system.

Claims

one . Compound of formula (I):

(I)

where

R ¹ is selected from H, CiCm alkyl or C1-C10 alkenyl and R ² is selected from CiC-ιο θ C1-C10 alkenyl optionally substituted by halogen,

or its pharmaceutically acceptable salts, solvates or isomers.

2. Compound according to claim 1 wherein R ¹ is H.

3. Compound according to claim 1 wherein R ¹ is CC ₄ alkyl.

4. Compound according to claim 3 wherein R ¹ is methyl.

5. Compound according to any of claims 1 to 4 wherein R ² is dC ₄ alkyl.

6. Compound according to claim 5 wherein R ² is methyl.

7. Compound according to any of claims 1 to 4 wherein R ² is - CH ₂ Br.

8. Compound of formula (I) that is selected from the list comprising:

■ 3-acetyl-4- (1 H-indole-3-yl) -1 H-pyrrole-2,5-dione ^■ 3-Acetyl-4- (1-methyl-1 H-indole-3-yl) -1 H-pyrrole-2,5-dione

^■ 3- (2-Bromoacetyl) -4- (1 H-indole-3-yl) -1 H-pyrrole-2,5-dione

or its pharmaceutically acceptable salts, solvates or isomers.

9. Use of a compound of formula (I) according to any of claims 1 to 8 for the manufacture of a medicament.

10. Use of a compound of formula (I) according to any of claims 1 to 8 for the manufacture of a medicament for the treatment and / or prevention of a disease selected from neurodegenerative diseases, inflammatory diseases, cancer, diabetes and processes regenerative

eleven . Use according to claim 10 wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson 's disease, amyotrophic lateral sclerosis, cerebral ischemia, Parkinsonism encephalitic post, dystonia, Tourette ^'s, periodic limb movements pathologies, restless leg syndrome , attention deficit hyperactivity disorders, Hungtinton's disease, progressive supranuclear palsy, Pick's disease, frontotemporal dementia, neuromuscular diseases.

12. Use according to claim 10 wherein the inflammatory disease is selected from Crohn's disease, ulcerative colitis, rheumatoid arthritis, atherosclerosis, vasculitis or multiple sclerosis.

13. Use according to claim 10 wherein the cancer is selected from glioblastoma, leukemia, lymphomas, lung, breast, prostate or colon cancer.

14. Use according to claim 10 wherein the diabetes is selected as non-dependent insulin type II diabetes.

1 5. Use of a compound of formula (I) according to any of claims 1 to 8 for the manufacture of a medicament to promote regenerative processes.

1 6. Use according to claim 15 where differentiation of the stem cells of the nervous system, the hematopoietic system, the bone system or the myocardium is involved in the regenerative process.

17. Pharmaceutical composition comprising a compound of formula (I) according to any one of claims 1 to 8 and a pharmaceutically acceptable carrier or adjuvant.

1 8. Composition according to claim 1 7 further comprising another active ingredient.