Dual g9a / GSK-3ß inhibitors

Abstract

The present invention refers to potent and specific G9a / GSK-3β inhibitors, pharmaceutical compositions comprising these compounds, and uses thereof in the treatment or prevention of diseases or conditions related to G9a / GSK-3β inhibitors overexpression.

Description

[0001] Dual G9a / GSK-3p inhibitors

[0002] TECHNICAL FIELD

[0003] The present invention refers to compounds which are potent and specific G9a / GSK3p inhibitors, as well as to pharmaceutical compositions comprising these compounds and uses thereof in the treatment or prevention of diseases or conditions related to G9a / GSK3p overexpression.

[0004] BACKGROUND ART

[0005] Alzheimer’s disease (AD) is the most common cause of dementia, making the disease a global health crisis that must be addressed. Until now, none of the approved AD treatments turned out to be a success. Of note, due to the advancement of our knowledge of AD, several mechanisms involved in the pathogenesis have been proposed to date and, more importantly, have been shown to overlap and influence each other, creating a complex pathogenic network. Thus, one of the reasons why single-target-directed drugs have failed to reach clinical trials is the pathological complexity found in AD. In fact, it could be better contrasted by the development of Multiple Targeting Directed Ligands (MTDLs), able to simultaneously modulate multiple targets involved in the onset of the pathology (R. R. Ramsay, et al., “A perspective on multi-target drug discovery and design for complex diseases”, Clin. Transl. Med. 2018, 7:3). In addition, MTDLs may improve both efficacy and safety relative to drugs that address only a single target. Moreover, as a new strategy in drug discovery for AD, compared to combination therapies, multifunctional molecules avoid drug-drug interactions, off-target adverse effects, poor patient compliance, and high development costs compared to combination therapies.

[0006] Multiple lines of evidence suggest that epigenetic alterations and tau pathology are two of the crucial causes of AD (B. Maloney, et al., “Epigenetics of dementia: understanding the disease as a transformation rather than a state””, Lancet Neurol. 2016, 15, 760-774; K. Li, et al., “Synaptic Dysfunction in Alzheimer’s Disease: A , Tau, and Epigenetic Alterations”, Mol. Neurobiol. 2018, 55, 3021-3032). Strikingly, overexpression of methyltransferase G9a and glycogen synthase kinase (GSK-3P) serve as drivers of cognitive impairment, leading to synaptic plasticity reduction, autophagy dysfunction, increasing Tau pathology, OS, and neuroinflammation collectively contributing to and / or resulting in age-related cognitive decline and neurodegenerative disease, particularly AD.

[0007] G9a and its repressive mark (H3k9me2) increase with age and produce memory impairment. In fact, H3k9me2 is related to the repression of genes implicated in synaptic plasticity, learning, and memory formation (A. Bellver-Sanchis A, et al., “G9a Inhibition Promotes Neuroprotection through GMFB Regulation in Alzheimer's Disease”, Aging Dis. 2024, 15, 311-337). In addition, G9a and its repressive mark (H3k9me2) induce autophagic dysfunction and oxidative stress. Furthermore, known G9a inhibitors tend to produce high toxicity, low efficacy in vivo and in vitro studies, and poor brain-blood barrier (BBB) permeability (A. Jana, et al. “3D QSAR pharmacophore based lead identification of G9a lysine methyltransferase towards epigenetic therapeutics”, J. Biomol. Struct. Dyn. 2023, 41 , 8635-8653).

[0008] On the other hand, GSK-3P increases with age, is overexpressed in AD patients, modulates tau hyperphosphorylation, induces AB toxicity and neuroinflammation, and promotes loss of memory consolidation, neurogenesis, and synaptic plasticity (E. Lauretti, et al., “Glycogen synthase kinase-3 signaling in Alzheimer's disease”, Biochim. Biophys. Acta Mol. Cell Res. 2020, 1867, 118664). GSK-3p inhibitors are usually associated with safety problems and have shown limited efficacy in clinical trials (S. M. Arciniegas Ruiz, et al., “Glycogen Synthase Kinase-3 Inhibitors: Preclinical and Clinical Focus on CNS-A Decade Onward”, Front. Mol. Neurosci. 2022, 21 :792364).

[0009] The present invention provides dual G9a / GSK-3p inhibitors capable of modulating simultaneously epigenetic alterations and tau pathology, two crucial causes of AD. Such dual inhibitors can reduce adverse effects and safety / toxicity problems, among other things. Therefore, our dual approach is an innovative and promising multifaceted therapeutic strategy for AD treatment and other G9a / GSK-3p-mediated disorders.

[0010] SUMMARY OF THE INVENTION

[0011] The present invention in a first aspect, refers to a compound of formula (I), or a salt, solvate or stereoisomer thereof: wherein: R is one or more radicals independently selected from the group consisting of: H; OH; CN; fluorine; chlorine; (Ci-Cw)haloalkyl; -0-(Ci-Cio)alkyl; (Ci-Cio)alkyl; -O-(Ci- Cw)haloalkyl; SF5; S(O)2R’I; NR^R’s; CONR^R’s; and COOR’e;

[0012] R'i, R'2, R’3, R’4, R’s and R’e, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (C3-C8)cycloalkyl;

[0013] X represents O or S;

[0014] Z represents O or NH; and

[0015] R1is selected from the group consisting of: a (Ci-Cw)alkyl; or a 4-halophenyl ring; for use in preventing or treating a G9a / GSK-3p-mediated disorder in a subject in need thereof, particularly by inhibiting GSK-3P and G9a; with the proviso that formula (I), or a salt, solvate or stereoisomer thereof, preferably does not contain any one of the following compounds selected from the group consisting of: Ethyl 4-(4-chlorophenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-p-tolyl-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Propyl 4-(3-chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-phenyl-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-m-tolyl-1 ,2,3,4- tetrahydropynmidme-5-carboxylate; Ethyl 4-(3-methoxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-methoxyphenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-fluorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-fluorophenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(2-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-(3-(trifluoromethoxy)phenyl)-

[0016] 1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Isobutyl 4-(3-chlorophenyl)-6-methyl-2-oxo-

[0017] 1.2.3.4-tetrahydropyrimidine-5-carboxylate; nd Ethyl 4-(3-cyanophenyl)-6-methyl-2-oxo-

[0018] 1.2.3.4-tetrahydropyrimidine-5-carboxylate.

[0019] In a preferred embodiment, the compound of formula (I) is characterized by:

[0020] R being selected from the group consisting of: H; fluorine; chlorine; (Ci-Cw)haloalkyl; - O-(Ci-C )alkyl; (Ci-Cw)alkyl and -O-(Ci-Cw)haloalkyl;

[0021] X represents O or S;

[0022] Z represents O or NH; and

[0023] R1a (Ci-C4)alkyl; or a 4-halophenyl. In another preferred embodiment, the compound of formula (I) is characterized by:

[0024] R being selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; - O-(Ci-C4)alkyl; (Ci-C4)alkyl and -O-(Ci-C4)haloalkyl;

[0025] X represents O or S;

[0026] Z represents O or NH; and

[0027] R1a (Ci-C4)alkyl; or a 4-halophenyl ring.

[0028] In another preferred embodiment, the compound is of formula (II), or a salt, solvate or stereoisomer thereof: wherein:

[0029] R is one or more radicals independently selected from the group consisting of: H; OH; CN; fluorine; chlorine; (Ci-Cw)haloalkyl; -0-(Ci-Cio)alkyl; (Ci-Cio)alkyl; -O-(Ci- Cw)haloalkyl; SF5; S(O)2R’I; NR^R’s; CONR^R’s; and COOR’e;

[0030] R'i, R'2, R’3, R’4, R’s and R’e, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (C3-C8)cycloalkyl; and

[0031] X represents O or S; with the proviso that formula (II), or a salt, solvate or stereoisomer thereof, does preferably not contain any one of the following compounds selected from the group consisting of: Ethyl 4-(4- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2- oxo-4-p-tolyl4,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-phenyl-

[0032] 1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-m-tolyl-1 ,2,3,4 tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-methoxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-methoxyphenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-fluorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-fluorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(2-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-hydroxyphenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-(3-(trifluoromethoxy)phenyl)-

[0033] 1.2.3.4-tetrahydropyrimidine-5-carboxylate; and Ethyl 4-(3-cyanophenyl)-6-methyl-2-oxo-

[0034] 1.2.3.4-tetrahydropyrimidine-5-carboxylate.

[0035] Preferably, wherein

[0036] R is one or more radicals independently selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; -O-(Ci-C4)alkyl; (Ci-C4)alkyl and -O-(Ci- C4)haloalkyl.

[0037] More preferably, wherein

[0038] X represents S; and

[0039] R is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and CF3.

[0040] Still more preferably, the compound of formula (II) is selected from the group consisting of: a) Ethyl 6-methyl-4-phenyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T1); b) Ethyl 4-(4-chlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T2); c) Ethyl 4-(4-methoxyphenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T3); d) Ethyl 6-methyl-2-thioxo-4-(p-tolyl)-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T4); e) Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T5); f) Ethyl 6-methyl-2-thioxo-4-[4-(trifluoromethyl)phenyl]-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T6); g) Ethyl 4-(2,4-dichlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T7); h) Ethyl 4-(2-chlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T8); and any salt, solvate or stereoisomer thereof.

[0041] In another preferred embodiment, the compound of formula (II) is characterized by:

[0042] X representing O; and

[0043] R is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and CF3. Preferably the compound of formula (II) is selected from the group consisting of: a) Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U5); b) Ethyl 6-methyl-2-oxo-4-[4-(trifluoromethyl)phenyl]-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U6); c) Ethyl 6-methyl-2-oxo-4-(2,4-dichlorophenyl)-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U7); and any salt, solvate or stereoisomer thereof.

[0044] In another preferred embodiment, the compound is of formula (III), or a salt, solvate or stereoisomer thereof: wherein:

[0045] R is one or more radicals independently selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; -O-(Ci-C4)alkyl; (Ci-C4)alkyl; and -O-(Ci- C4)haloalkyl.

[0046] Preferably, wherein

[0047] R is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and -OCF3.

[0048] More preferably, the compound of formula (III) is selected from the group consisting of: a) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-phenyl-1 ,2,3,4-tetrahydropyrimidine-5- carboxamide (CU1); b) A / , 4-Bis(4-chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxamide (CU2); c) / \ / -(4-Chlorophenyl)-4-(4-methoxyphenyl)-6-methyl-2-oxo-1, 2,3,4- tetrahydropyrimidine-5-carboxamide (CU3); d) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-(p-tolyl)-1 ,2,3,4-tetrahydropyrimidine-5- carboxamide (CU4); e) / \ / -(4-Chlorophenyl)-4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxamide (CU5); and f) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-[4-(trifluoromethoxy)phenyl]-1 , 2,3,4- tetrahydropyrimidine-5-carboxamide (CU6). any salt, solvate or stereoisomer thereof.

[0049] In a preferred embodiment, the G9a / GSK-3p-mediated disorder is a protein-aggregation disease and the compound is for use in the treatment and / or prevention of a proteinaggregation disease. Preferably, the protein-aggregation disease is Alzheimer’s disease, Parkinson’s disease or Huntington’s disease.

[0050] In a preferred embodiment, the G9a / GSK-3p-mediated disorder is a disease selected from the group consisting of Alzheimer’s disease, amyotrophic lateral sclerosis, age-related tau astrogliopathy, aortic amyloidosis, argyrophilic grain disease, British familial dementia, cardiac amyloidosis, cerebral amyloid angiopathy, chronic traumatic encephalopathy, corneal dystrophies, corticobasal degeneration, Creutzfeldt-Jakob disease, Danish familial dementia, Down syndrome, familial amyloidosis, familial corneal amyloidosis, fatal insomnia, frontotemporal dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, hereditary cerebral hemorrhage with amyloidosis, Huntington’s disease, inflammation- associated amyloidosis, kuru, Lewy bodies dementia, Mediterranean fever, Niemann-Pick disease type C, Parkinson’s disease, Pick’s disease, primary age-related tauopathy, progressive subcortical gliosis, progressive supranuclear palsy systemic amyloidosis, including transthyretin-associated amyloidosis and light-chain amyloidosis, subacute sclerosing panencephalitis and tuberous sclerosis. Preferably, the disease is Alzheimer’s disease.

[0051] DESCRIPTION OF EMBODIMENTS

[0052] Terms not specifically defined herein should be given the meanings that would be given to them by one of skill in the art in light of the disclosure and the context. As used in the specification, however, unless specified to the contrary, the following terms have the meaning indicated and the following conventions are adhered to.

[0053] Throughout the present specification and the accompanying clauses, the words "comprise" and variations such as "comprises", "comprising" are to be interpreted inclusively. That is, these words are intended to convey the possible inclusion of other elements or integers not specifically recited, where the context allows. The word “comprise” also includes the term “consists of”.

[0054] For the purposes of the present invention, any ranges given include both the lower and the upper end-points of the range.

[0055] As already stated, inventors have found that dual inhibition of G9a and GSK-3P has a significant effect on the contemporaneous neuroinflammation and amyloid pathology, two crucial early mechanisms of AD. Thus, inventors disclose here small-molecule compounds endowed with dual activity on both G9a and GSK-3p, along with suitable drug-like properties. These findings hold great potential to halt or delay AD progress and other pathological conditions describe herein.

[0056] Thus, in a first aspect the present invention provides the use of compounds of formula (I) (herein referred to as “compounds of the invention”) in preventing or treating an G9a / GSK-3p- mediated disorder in a subject in need thereof, particularly by inhibiting GSK-3P and G9a. or a salt, solvate or stereoisomer thereof; wherein:

[0057] R is one or more radicals independently selected from the group consisting of: H; OH;

[0058] CN; fluorine; chlorine; -(Ci-Cw)haloalkyl; -0-(Ci-Cio)alkyl; -(Ci-Cio)alkyl; -O-(Ci- Cw)haloalkyl; SFs; S(O)2 ’I; NR^R’a; CONR^R’s; and COOR’e;

[0059] R'i, R'2, R’3, R’4, R’5and R’e, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (Cs-Csjcycloalkyl;

[0060] X represents O or S;

[0061] Z represents O or NH; and

[0062] R1is selected from the group consisting of: a (Ci-Cw)alkyl; or a 4-halophenyl ring; with the proviso that formula (I), or a salt, solvate or stereoisomer thereof, does preferably not contain any one of the following compounds selected from the group consisting of: Ethyl 4-(4- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2- oxo-4-p-tolyl-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Propyl 4-(3-chlorophenyl)-6-methyl-

[0063] 2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-phenyl-1 ,2,3,4 tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-m-tolyl-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-methoxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-methoxyphenyl)-6-methyl-2-oxo-1 ,2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-fluorophenyl)-6-methyl-2-oxo-1 ,2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-fluorophenyl)-6-methyl-2-oxo-1 , 2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(2-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-(3-(trifluoromethoxy)phenyl)' l,2,3,4-tetrahydropyrimidine-5-carboxylate; Isobutyl 4-(3-chlorophenyl)-6-methyl-2-oxO'

[0064] 1.2.3.4-tetrahydropyrimidine-5-carboxylate; and Ethyl 4-(3-cyanophenyl)-6-methyl-2-oxo-

[0065] 1.2.3.4-tetrahydropyrimidine-5-carboxylate.

[0066] In the context of the invention, the term "alkyl" refers to a straight or branched hydrocarbon chain radical containing no unsaturation, and which is attached to the rest of the molecule by a single bond. Typical alkyl groups have from 1 to about 10, 1 to about 8, or 1 to about 6 carbon atoms, e. g., methyl, ethyl, n-propyl, / -propyl, n-butyl, f-butyl, n-pentyl, etc. If substituted by cycloalkyl, it corresponds to a "cycloalkylalkyl" radical, such as cyclopropyl methyl. If substituted by aryl, it corresponds to an "arylalkyl" radical, such as benzyl, benzhydryl or phenethyl. If substituted by heterocyclyl, it corresponds to a "heterocyclylalkyl" radical.

[0067] In the context of the present invention, the term haloalkyl, such as in “(C1-C10)haloalkyl”, refers to an alkyl group, as defined above, having carbon atoms substituted with at least one halo or halogen groups. Preferably, in the specific case of “(C1-C10)haloalkyl”, the alkyl group contains at least 1 , and at most 10, carbon atoms substituted with at least one halo or halogen group, halogen being as defined herein. Examples of "C1-C10 haloalkyl" groups useful in the present invention include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, isobutyl and n-butyl substituted independently with one or more halo groups, e.g., fluoro, chloro, bromo and iodo. A particularly useful example of a (C1-C10)haloalkyl group is -CF3. It is noted that preferably, the invention refers to (C1-C6)haloalkyl, (C1-C4)haloalkyl and (C1-C2)haloalkyl groups.

[0068] In the context of the present invention, the term “-O-(C1-C10)haloalkyl” refers to the group RaO-, where Ra is a haloalkyl, preferably a C1-C10 haloalkyl, as defined above. An exemplary C1-C10 haloalkoxy group useful in the present invention includes, but is not limited to, trifluoromethoxy. The term “-O-(C1-C10)haloalkyl” can thus also be referred to as a C1 to C10 haloalkoxy group.

[0069] In the context of the invention, the term “alkoxy” refers to a -O-alkyl such as -0-(Ci-Cio)alkyl, where the alkyl group, such as (Ci-Cio)alkyl, is an alkyl as defined above and the term "C1- C10 alkoxy" or “-0-(Ci-Cio)alkyl” refers to the group -O-alkyl , where the alkyl group is a C1- C10 alkyl as defined above. Examples of "-0-(Ci-Cio)alkyl" groups useful in the present invention include, but are not limited to, methoxy, ethoxy, propyloxy, and isopropyloxy.

[0070] In the context of the invention, the term "halogen" or “halo” refers to bromo, chloro, iodo or fluoro.

[0071] In the context of the present invention, the term "salt" must be understood as any form of a compound used in accordance with this invention in which said compound is in ionic form or is charged and coupled to a counter-ion (a cation or anion) or is in solution. This definition also includes quaternary ammonium salts and complexes of the active molecule with other molecules and ions, particularly, complexes formed via ionic interactions. The definition includes in particular physiologically acceptable salts; this term must be understood as equivalent to "pharmacologically acceptable salts" or "pharmaceutically acceptable salts".

[0072] In the context of the present invention, the term "pharmaceutically acceptable salts" means any salt that is tolerated physiologically (normally meaning that it is not toxic, particularly, as a result of the counter-ion) when used in an appropriate manner for a treatment, applied or used, particularly, in humans and / or mammals. These physiologically acceptable salts may be formed with cations or bases and, in the context of this invention, are understood to be salts formed by at least one compound used in accordance with the invention -normally an acid (deprotonated)- such as an anion and at least one physiologically tolerated cation, preferably inorganic, particularly when used in humans and / or mammals. Salts with alkali and alkali earth metals are preferred particularly, as well as those formed with ammonium cations (NH4+). Preferred salts are those formed with (mono) or (di)sodium, (mono) or (di)potassium, magnesium or calcium. These physiologically acceptable salts may also be formed with anions or acids and, in the context of this invention, are understood as being salts formed by at least one compound used in accordance with the invention - normally protonated, for example in nitrogen - such as a cation and at least one physiologically tolerated anion, particularly when used on humans and / or mammals. This definition specifically includes in the context of this invention a salt formed by a physiologically tolerated acid, i.e., salts of a specific active compound with physiologically tolerated organic or inorganic acids - particularly when used on humans and / or mammals. Examples of this type of salts are those formed with: hydrochloric acid, hydrobromic acid, sulphuric acid, methanesulfonic acid, formic acid, acetic acid, oxalic acid, succinic acid, malic acid, tartaric acid, mandelic acid, fumaric acid, lactic acid or citric acid.

[0073] In the context of the present invention, the term "solvate" should be understood as meaning any form a compound in accordance with the invention in which said compound is bonded by a non-covalent bond to another molecule (normally a polar solvent), including especially hydrates and alcoholates, like for example, methanolate. A preferred solvate is the hydrate.

[0074] The term "prodrug" is used in its broadest sense and encompasses those derivatives that are converted in vivo to the compounds of the invention. Examples of prodrugs include, but are not limited to, derivatives and metabolites of the compounds of formula (I) that include biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogues. Preferably, prodrugs of compounds with carboxyl functional groups are the lower alkyl esters of the carboxylic acid. The carboxylate esters are conveniently formed by esterifying any of the carboxylic acid moieties present on the molecule. Prodrugs can typically be prepared using well-known methods.

[0075] Any compound of formula (I) referred to herein is intended to represent such specific compound as well as certain variations or forms. In particular, compounds referred to herein may have asymmetric centres and therefore exist in different enantiomeric or diastereomeric forms. Thus, any given compound of formula (I) referred to herein is intended to represent any one of a racemate, one or more enantiomeric forms, one or more diastereomeric forms, and mixtures thereof. Likewise, stereoisomerism or geometric isomerism about the double bond is also possible, therefore in some cases the molecule could exist as (E)-isomer or (Z)-isomer (trans and cis isomers). If the molecule contains several double bonds, each double bond will have its own stereoisomerism, that could be the same as, or different to, the stereoisomerism of the other double bonds of the molecule. Furthermore, compounds referred to herein may exist as atropisomers. All the stereoisomers including enantiomers, diastereoisomers, geometric isomers and atropisomers of the compounds referred to herein, and mixtures thereof, are considered within the scope of the present invention.

[0076] Furthermore, any compound of formula (I) referred to herein may exist as tautomer. Specifically, the term tautomer refers to one of two or more structural isomers of a compound that exist in equilibrium and are readily converted from one isomeric form to another.

[0077] In one embodiment of the first aspect of the invention, the compound of formula (I) is characterized by R being selected from the group consisting of: H; fluorine; chlorine; (Ci-Cw)haloalkyl; - O-(Ci-C )alkyl; -(Ci-Cw)alkyl and -O-(Ci-Cw)haloalkyl;

[0078] X represents O or S;

[0079] Z represents O or NH; and

[0080] R1a (Ci-C4)alkyl; or a 4-halophenyl ring.

[0081] In another embodiment of the first aspect of the invention, the compound of formula (I) is characterized by

[0082] R being selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; - O-(Ci-C4)alkyl; (Ci-C4)alkyl and -O-(Ci-C4)haloalkyl;

[0083] X represents O or S;

[0084] Z represents O or NH; and

[0085] R1a (Ci-C4)alkyl; or a 4-halophenyl ring.

[0086] In yet another embodiment of the first aspect of the invention, the compound of formula (I) is characterized by

[0087] R being selected from the group consisting of: H; fluorine; chlorine; (Ci-C2)haloalkyl such as CF3 or CHF2; -O-(Ci-C2)alkyl such as a methoxy group; (Ci-C2)alkyl such as a methyl group and -O-(Ci-C2)haloalkyl such as a OCF3;

[0088] X represents O or S;

[0089] Z represents O or NH; and

[0090] R1a (Ci-C2)alkyl or a 4-halophenyl ring.

[0091] In another embodiment of the first aspect of the invention, the compound is of formula (II), or a salt, solvate or stereoisomer thereof; wherein: R is one or more radicals independently selected from the group consisting of: H; OH; CN; fluorine; chlorine; -(Ci-Cw)haloalkyl; -0-(Ci-Cio)alkyl; -(Ci-Cw)alkyl; -O-(Ci- Cw)haloalkyl; SFs; S(O)2R’I; NR^R’s; CONR^R’s; and COOR’e;

[0092] R'i, R'2, R’3, R’4, R’5and R’e, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (C3-C8)cycloalkyl; and

[0093] X represents O or S; with the proviso that formula (II), or a salt, solvate or stereoisomer thereof, does preferably not contain any one of the following compounds selected from the group consisting of: Ethyl 4-(4- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2- oxo-4-p-tolyl-l,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-phenyl- l,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-m-tolyl-1 ,2,3,4 tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-methoxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-methoxyphenyl)-6-methyl-2-oxo-1 ,2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-fluorophenyl)-6-methyl-2-oxo-1 ,2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-fluorophenyl)-6-methyl-2-oxo-1 , 2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(2-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4' tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-(3-(trifluoromethoxy)phenyl)'

[0094] 1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; and Ethyl 4-(3-cyanophenyl)-6-methyl-2-oxo-

[0095] 1 ,2,3,4-tetrahydropyrimidine-5-carboxylate.

[0096] Preferably, wherein in the compound of formula (II):

[0097] R is one or more radicals independently selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; -O-(Ci-C4)alkyl; (Ci-C4)alkyl and -O-(Ci- C4)haloalkyl.

[0098] Preferably, wherein in the compound of formula (II):

[0099] R is one or more radicals independently selected from the group consisting of: H; fluorine; chlorine; (Ci-C2)haloalkyl; -O-(Ci-C2)alkyl; (Ci-C2)alkyl and -O-(Ci- C2)haloalkyl.

[0100] More preferably, wherein in the compound of formula (II):

[0101] X represents S; and

[0102] R is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and CF3. More preferably, wherein the compound of formula (II) is selected from the group consisting of: a) Ethyl 6-methyl-4-phenyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T1); b) Ethyl 4-(4-chlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T2); c) Ethyl 4-(4-methoxyphenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T3); d) Ethyl 6-methyl-2-thioxo-4-(p-tolyl)-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T4); e) Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T5); f) Ethyl 6-methyl-2-thioxo-4-[4-(trifluoromethyl)phenyl]-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T6); g) Ethyl 4-(2,4-dichlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T7); h) Ethyl 4-(2-chlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T8); and any salt, solvate or stereoisomer thereof.

[0103] In another embodiment of the first aspect of the invention, the compound is of formula (II) is characterized by:

[0104] X representing O; and

[0105] R being one or more radicals independently selected from the group consisting of: H; Cl; -OCH3; CH3and CF3.

[0106] More preferably, wherein the compound of formula (II) is selected from the group consisting of: a) Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U5); b) Ethyl 6-methyl-2-oxo-4-[4-(trifluoromethyl)phenyl]-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U6); c) Ethyl 6-methyl-2-oxo-4-(2,4-dichlorophenyl)-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U7); and any salt, solvate or stereoisomer thereof.

[0107] In another embodiment of the first aspect of the invention, the compound is of formula (III), or a salt, solvate or stereoisomer thereof; wherein:

[0108] R is one or more radicals independently selected from the group consisting of: H; fluorine; chlorine; -(Ci-C4)haloalkyl; -O-(Ci-C4)alkyl; -(Ci-C4)alkyl; and -O-(Ci- C4)haloalkyl.

[0109] Preferably, wherein in the compound of formula (III):

[0110] R is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and -OCF3.

[0111] More preferably, wherein the compound of formula (III) is selected from the group consisting of: a) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-phenyl-1 ,2,3,4-tetrahydropyrimidine-5- carboxamide (CU1); b) A / , 4-Bis(4-chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxamide (CU2); c) / \ / -(4-Chlorophenyl)-4-(4-methoxyphenyl)-6-methyl-2-oxo-1, 2,3,4- tetrahydropyrimidine-5-carboxamide (CU3); d) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-(p-tolyl)-1 ,2,3,4-tetrahydropyrimidine-5- carboxamide (CU4); e) / \ / -(4-Chlorophenyl)-4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1, 2,3,4- tetrahydropyrimidine-5-carboxamide (CU5); and f) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-[4-(trifluoromethoxy)phenyl]-1 , 2,3,4- tetrahydropyrimidine-5-carboxamide (CU6); and any salt, solvate or stereoisomer thereof. It is noted that G9a and GLP (G9a like protein) are major enzymes that catalyze the mono- and dimethylation of a lysine residue at position 9 of histone H3 (H3K9me1 and H3K9me2). These enzymes are also known as EHMT2 and EHMT1 (euchromatin histone-lysine N- methyltransferases 2 and 1).

[0112] H3K9me2 is an epigenetic mark related to transcriptional repression. G9a and GLP are involved in epigenetic transcriptional repression through H3K9me2. The inhibition of G9a has been widely reported as useful for the control of biological processes, such as cell proliferation and cell differentiation, mediated by transcriptional repression by H3K9me2.

[0113] On the other hand, Glycogen synthase kinase- 3 (GSK-3) is a serine / threonine kinase widely expressed and active in unstimulated tissues. GSK-3 exists in two isoforms, GSK-3a and GSK- 3p, having 98% homology in their catalytic domain and difference in their C-terminal and N- terminal sequences. The GSK-3P isoform is predominantly expressed in the brain. It is one of the predominant kinases which cause tau hyperphosphorylation, destabilizes the microtubules, and leads to the formation of NFTs. APP and presenilin 1 (PS1) are also GSK- 3p substrates. GSK-3P is therefore implicated in controlling the synthesis of Ap as well. By decreasing BACE-1 -induced cleavage of APP through the Nuclear factor kappa light chain enhancer of activated B cells (NF-KB) pathway, GSK-3P inhibition is said to play a role in reducing the formation of Ap plaques. Inhibition of GSK-3P in cultured neurons also tends to reduce Ap-induced neurotoxicity. The role of Ap has also been hypothesized in developing filaments similar to paired helical filaments (PHF), an important part of NFTs, one of AD's pathogenic characteristics. Thus, one of the promising disease-modifying strategies for combating AD involves the inhibition of GSK3p.

[0114] The present invention provides compounds which are potent G9a and GSK3P inhibitors.

[0115] As used herein, the term “inhibit” refers to a reduction or decrease in a quality or quantity, compared to a baseline.

[0116] In one embodiment, the G9a and / or GSK3p-mediated disorder is associated with overexpression of G9a. G9a overexpression has already been reported in numerous cancers such as hematologic, breast, gastric, ovarian, prostate, lung, colorectal, liver, urinary bladder, and brain cancers has sparked interest in it for therapeutic development (S. H. Barghout, et al., “Chemical biology and pharmacology of histone lysine methylation inhibitors”, Biochim. Biophys. Acta Gene Regul. Meeh. 2022, 1865:194840); as well as involved in immune system activation (S. Scheer, et al., “The Lysine Methyltransferase G9a in immune Cell Differentiation and Function”, Front. Immunol. 2017, 8:429); in memory function (K. K. L. Pang, et al., “Epigenetics and memory: Emerging role of histone lysine methyltransferase G9a / GLP complex as bidirectional regulator of synaptic plasticity”, Neurobiol. Learn. Mem. 2019, 159:1- 5); or Prader-Willi syndrome (Y. Kim, et al., “Epigenetic therapy of Prader-Willi syndrome”, Transl. Res., 2019, 208, 105-118).

[0117] It is noted that the compound of formula (I) or any of the compounds described in the present invention, in the context of the invention, can be administered in combination with any other therapeutic agent for preventing or treating the disorder, including G9a and / or GSK3P inhibitors, where the combination of the drugs together is safer or more effective than either drug alone. Such other drug(s) may be administered, by a route and in an amount commonly used therefore, contemporaneously or sequentially with the composition. When a compound of Formula I is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and the compound of formula I is preferred. However, the combination therapy may also include therapies in which the compound of Formula I and one or more other drugs are administered on different overlapping schedules. It is also contemplated that when used in combination with one or more other active ingredients, the compounds of the present invention and the other active ingredients may be used in lower doses than when each is used singly. Accordingly, the pharmaceutical compositions of the present invention include those that contain one or more other active ingredients, in addition to a compound of Formula I. The above combinations include combinations of a compound of the present invention not only with one other active compound, but also with two or more other active compounds. For example, the compound of Formula I can be combined with a variety of different anti-neurodegenerative drugs, including those targeting Alzheimer’s disease, Parkinson’s disease and other related conditions.

[0118] Further, the compound of Formula I can be combined with the following, but not limited to, anticholinergic antiparkinson agents such as diphenhydramine benztropine, trihexyphenidyl, procyclidine or biperiden, dopaminergic antiparkinsonism agents such as ramipexole, rotigotine, selegiline, carbidopa I levodopa, bromocriptine, rasagiline, entacapone, ropinirole, pramipexole or safinamide, or miscellaneous antiparkinson agents such as istradefylline. In addition, the compound of Formula I can also be combined with the following, but not limited to, atypical antipsychotic agents such as rexpiprazole; cholinesterase inhibitors such as donepezil, galantamine, or rivastigmine; NMDA antagonists such as memantine; or diseasemodifying immunotherapy such as lecanemab.

[0119] Likewise, compounds of the present invention may be used in combination with other drugs that are used in the prevention, treatment, control, amelioration, or reduction of risk of the diseases or conditions for which compounds of the present invention are useful as described above. Such other drugs may be administered, by a route and in an amount commonly used therefore, contemporaneously or sequentially with a compound of the present invention. When a compound of the present invention is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound of the present invention is preferred. Accordingly, the pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients, in addition to a compound of the present invention.

[0120] The weight ratio of the compound of the compound of the present invention to the second active ingredient may be varied and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when a compound of the present invention is combined with another agent, the weight ratio of the compound of the present invention to the other agent will generally range from about 1000:1 to about 1 :1000, preferably about 200:1 to about 1 :200. Combinations of a compound of the present invention and other active ingredients will generally also be within the aforementioned range, but in each case, an effective dose of each active ingredient should be used. In such combinations the compound of the present invention and other active agents may be administered separately or in conjunction. In addition, the administration of one element may be prior to, concurrent to, or subsequent to the administration of other agent(s).

[0121] The skilled person will routinely adjust the amounts and appropriate route of administration according to their expertise and the particular situation of the subject.

[0122] In another embodiment, the G9a and / or GSK3p-mediated disorder is a protein-aggregation disease and the compound of formula (I) or any of the compounds described in the present invention, are for use in the treatment and / or prevention of a protein-aggregation disease. The protein-aggregation disease is preferably Alzheimer’s disease, Parkinson’s disease or Huntington’s disease, more preferably Alzheimer’s disease.

[0123] In another embodiment, the disease is selected from the group consisting of Alzheimer’s disease, amyotrophic lateral sclerosis, age-related tau astrogliopathy, aortic amyloidosis, argyrophilic grain disease, British familial dementia, cardiac amyloidosis, cerebral amyloid angiopathy, chronic traumatic encephalopathy, corneal dystrophies, corticobasal degeneration, Creutzfeldt-Jakob disease, Danish familial dementia, Down syndrome, familial amyloidosis, familial corneal amyloidosis, fatal insomnia, frontotemporal dementia, Gerstmann-Straussler-Scheinker disease, globular glial tauopathy, hereditary cerebral hemorrhage with amyloidosis, Huntington’s disease, inflammation-associated amyloidosis, kuru, Lewy bodies dementia, Mediterranean fever, Niemann-Pick disease type C, Parkinson’s disease, Pick’s disease, primary age-related tauopathy, progressive subcortical gliosis, progressive supranuclear palsy systemic amyloidosis, including transthyretin-associated amyloidosis and light-chain amyloidosis, subacute sclerosing panencephalitis and tuberous sclerosis. In another embodiment, the disorder is a blood disorder, such as sickle cell anemia or b- thalassemia or hematological cancer.

[0124] In another embodiment the disorder is a cancer, such as lymphoma, leukemia, melanoma, breast cancer, ovarian cancer, hepatocellular carcinoma, prostate carcinoma, lung cancer, brain cancer, or hematological cancer.

[0125] In another embodiment the disorder is an immune-mediated disease.

[0126] In another embodiment the compounds are used for reducing substance (alcohol) consumption by a subject.

[0127] In another embodiment the disorder is an imprinting disorder, such as Prader-Willi syndrome, transient neonatal diabetes mellitus, Silver-Russell syndrome, Albright hereditary osteodystrophy, pseudohypoparathyroidism, Birk-Barel mental retardation, Beckwith- Wiedemann syndrome, Temple syndrome, Kagami-Ogata syndrome, Angelman syndrome, precocious puberty, Schaaf-Yang syndrome, sporadic pseudohypoparathyroidism lb, or maternal uniparental disomy of chromosome 20 syndrome.

[0128] In a further aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the compounds of the invention (I) as well as solvates, salts or stereoisomers thereof, as defined above. All the embodiments provided above, under the first aspect of the invention, concerning the compounds of formula (I), are also embodiments of the pharmaceutical composition of the invention.

[0129] By “therapeutically effective amount”, it is understood the amount of the compound(s) that, when administered, is sufficient to prevent development of, or alleviate to some extent, one or more of the symptoms of the disease which is addressed.

[0130] The precise therapeutic dose of the component(s), as well as the amount of the compound(s) of the invention, may depend on several variables. Some of these would be: route of administration, time of drug release (e.g., instant or extended), administration schedule, pain severity, condition of the patient, and the like.

[0131] The pharmaceutical compositions can be prepared as a liquid, semi-solid or solid dosage 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, or administered as such.

[0132] These compositions can be prepared with the aid of conventional means, devices, methods or processes known in the art. Pharmaceutically acceptable adjuvants, vehicles or excipients which may be used in such compositions are adjuvants, vehicles or excipients known to those skilled in the art or commonly used in the preparation of therapeutic compositions, which may be selected, for example, from the group consisting of excipients, fillers, solvents, diluents, surfactants, colorants, preservatives, disintegrants, sliding agents, lubricants, flavoring agents or binders.

[0133] The term "pharmaceutically acceptable" refers to pharmaceutically acceptable materials, compositions or vehicles. Each component must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the pharmaceutical composition. It must also be suitable for use in contact with the tissue or organ of humans without excessive toxicity, irritation, allergic response, immunogenicity or other problems or complications commensurate with a reasonable benefit / risk ratio in animals and, particularly, in humans.

[0134] As used herein, the term “animal” or “subject” or “patient” shall refer to a vertebrate animal. Such animals include both domestic animals; for example, livestock, laboratory animals and household pets, and non-domestic animals such as wildlife. In one embodiment, the animal is a vertebrate. In a particular embodiment the animal is a domestic mammal or a human.

[0135] The selection of physiologically compatible adjuvants or the number of adjuvants to be used depends on the form of administration of the pharmaceutical composition, i.e., oral, subcutaneous, parenteral, intravenous, intraperitoneal, intradermal, intramuscular, intranasal, buccal, rectal, otic or intratympanic. Preparations in the form of tablets, dragees, capsules, granules, pills, drops, in particular otic drops, juices or syrups are preferably suitable for oral administration; solutions, suspensions, easily reconstitutable dry preparations or also sprays are preferably suitable for parenteral, topical or inhalation administration. The compounds in accordance with the invention used in the pharmaceutical composition in accordance with the invention in a depot, in a dissolved form or in a dressing, or if appropriate having added other agents favoring penetration into the skin, are preparations suitable for percutaneous administration. The preparation forms administrable orally or percutaneously can also release the respective compound according to the invention in a delayed form.

[0136] For instance, for oral administration in the form of a tablet or capsule, the active drug components can be combined with an oral, non-toxic, pharmaceutically acceptable, inert carrier such as lactose, starch, sucrose, glucose, methyl cellulose, magnesium stearate, dicalcium phosphate, calcium sulphate, mannitol, sorbitol and the like; for oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Moreover, when desired or necessary, suitable binders, lubricants, disintegrating agents, and colouring agents can also be incorporated into the mixture. Suitable binders include starch, gelatine, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

[0137] Gelatine capsules contain the active ingredient and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be manufactured as sustained release products to provide for continuous release of medication over a period of hours. Compressed tablets can be sugar coated or film coated to mask any unpleasant taste and protect the tablet from the atmosphere, or enteric coated for selective disintegration in the gastrointestinal tract.

[0138] Liquid dosage forms for oral administration can contain colouring and flavouring to increase patient acceptance.

[0139] The dosage administered of the pharmaceutical composition will, of course, vary depending on the use and known factors such as the age, health, and weight of the recipient; nature and extent of symptoms, concurrent treatments, if any, frequency of treatment, and the effect desired. The recipient may be any type of mammal, but is preferably a human.

[0140] To those skilled in the art, other objects, advantages or features of the invention will be apparent in part from the description or in part from the practice of the invention. The following examples are provided by way of illustration or are not intended to be limiting of the present invention.

[0141] EXAMPLES

[0142] Analytical methods:

[0143] Melting points were determined in open capillary tubes with a MFB 595010 M Gallenkamp melting point apparatus. Infrared (IR) spectra were run on a Perkin-Elmer Spectrum RX I spectrophotometer (using the attenuated total reflectance (ATR) technique). Absorption values are expressed as wavenumbers (cm-1); only significant absorption bands are given. Preparative normal phase chromatography was performed on a CombiFlash Rf 150 (Teledyne Isco) with pre-packed RediSep Rf silica gel cartridges. Thin-layer chromatography was performed with aluminum-backed sheets with silica gel 60 F254 (Merck, ref 1 .05554 or Sigma- Aldrich, ref 60805), and spots were visualized with UV light, 1 % aqueous solution of KMnO4 and / or ninhydrin. High-resolution mass spectrometry (HRMS) analyses were performed with an LC / MSD TOF Agilent Technologies spectrometer. Analytical grade solvents were used for crystallization, while pure for synthesis solvents were used in the reactions, extractions, and column chromatography. Several compounds described in the present invention (T1, T2, T3, T4, T5, T6, T7, T8, U1, U2, U3, U4, U5, U6, U7, CU1 and CU4) were either commercially available or were synthetized following procedures known in the art (CN101463011A, CN1803777A, W02007105233A2).

[0144] Ethyl 6-methyl-4-phenyl-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (T1). IR 3327, 3170, 3108, 1667, 1573, 1464, 1370, 1327, 1283, 1194, 1176, 1118, 1027, 1001 , 821 , 759, 723, 692, 651 cm’1.

[0145] Ethyl 4-(4-chlorophenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (T2). IR 3327, 3173, 2984, 1670, 1573, 1465, 1281 , 1197, 1178, 1120, 1093, 1014, 805, 760, 746, 646 cm’1.

[0146] Ethyl 4-(4-methoxyphenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydropyrimidine-5- carboxylate (T3). IR 3312, 3168, 1666, 1609, 1573, 1509, 1462, 1269, 1251 , 1194, 1181 , 1170, 1121 , 1109, 1028, 835, 819, 765, 654 cm’1.

[0147] Ethyl 6-methyl-2-thioxo-4-(p-tolyl)-1,2,3,4-tetrahydropyrimidine-5-carboxylate (T4). IR 3323, 3170, 1671 , 1574. 1463, 1327, 1185, 1174, 1117, 786, 759, 763, 650 cm’1.

[0148] Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydropyrimidine-5- carboxylate (T5). IR: 3315, 3173, 1666, 1576, 1466, 1199, 1181 , 1112, 1030, 821 , 747 cm’1.

[0149] Ethyl 6-methyl-2-thioxo-4-[4-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydropyrimidine-5- carboxylate (T6). IR: 3320, 3170, 3103, 2993, 1673, 1574, 1467, 1372, 1327, 1285, 1200, 1170, 1113, 1066, 1017, 857, 816, 760. 746, 609, 640 cm’1.

[0150] Ethyl 4-(2,4-dichlorophenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydropyrimidine-5- carboxylate (T7). IR 3175, 2979, 1709, 1649, 1560, 1463, 1315, 1193, 1176, 1097, 850, 824, 754, 644 cm’1.

[0151] Ethyl 4-(2-chlorophenyl)-6-methyl-2-thioxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (T8). IR 3175, 2984, 1707, 1651 , 1572, 1471 , 1313, 1201 , 1179, 1090, 1036, 727, 643 cm’1.

[0152] Ethyl 6-methyl-2-oxo-4-phenyl-1,2,3,4-tetrahydropyrimidine-5-carboxylate (U1). IR 3237, 3112, 1722, 1698, 1644, 1463, 1418, 1385, 1289, 1270, 1218, 1087, 1027, 955, 878, 755, 697, 660, 630 cm’1.

[0153] Ethyl 4-(4-chlorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate

[0154] (U2). IR 3232, 3113, 1700, 1645, 1459, 1419, 1366, 1290, 1217, 1086, 1010, 780, 682, 629 cm'1.

[0155] Ethyl 4-(4-methoxyphenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (U3). IR 3236, 3113, 1724, 1704, 1651 , 1514, 1455, 1278, 1257, 1222, 1177, 1086, 1031 , 781 , 659 cm’1. Ethyl 6-methyl-2-oxo-4-(p-tolyl)-1,2,3,4-tetrahydropyrimidine-5-carboxylate (U4). IR

[0156] 3239, 3113, 2984, 1701 , 1648, 1459, 1286, 1221 , 1088, 787 cm’1.

[0157] Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxylate (U5). IR 3350, 3107, 2974, 1694, 1645, 1461 , 1320, 1297, 1227, 1099, 872, 799, 721 , 663 crrr 1

[0158] Ethyl 6-methyl-2-oxo-4-[4-(trifluoromethyl)phenyl]-1,2,3,4-tetrahydropyrimidine-5- carboxylate (U6). IR 3242, 3113, 1701 , 1644, 1333, 1289, 1221 , 1126, 1091 , 1069, 1018, 790, 778, 715, 665 cm’1.

[0159] Ethyl 6-methyl-2-oxo-4-(2,4-dichlorophenyl)-1,2,3,4-tetrahydropyrimidine-5-carboxylate (U7). IR 3359, 3103, 2966, 1695, 1643, 1455, 1323, 1297, 1226, 1095, 852, 816, 798, 653 crrr 1

[0160] A / -(4-Chlorophenyl)-6-methyl-2-oxo-4-phenyl-1,2,3,4-tetrahydropyrimidine-5- carboxamide (CU1). IR 3401, 3271 , 1711 , 1669, 1629, 1508, 1492, 1396, 1321 , 1243, 1089,

[0161] 1013, 819, 748, 697, 682, 656, 624 cm’1.

[0162] A / -(4-Chlorophenyl)-6-methyl-2-oxo-4-(p-tolyl)-1,2,3,4-tetrahydropyrimidine-5- carboxamide (CU4). IR 3397, 3278, 1707, 1668, 1628, 1592, 1508, 1397, 1244, 1231 , 1090,

[0163] 1014, 820, 764, 717, 682, 638, 648, 626 cm’1.

[0164] General procedure for the synthesis of compounds CU2, CU3, CU5 and CU6.

[0165] A mixture of the required aldehyde (4-chlorobenzaldehyde for CU2; p-anisaldehyde for CU3; 3,4-dichlorobenzaldehyde for CU5 and 4-(trifluoromethoxy)benzaldehyde for CU6) (1 .0 mmol), / V-(4-chlorophenyl)-3-oxobutanamide (1 mmol), urea (1.5 mmol) and Cu(OTf)2 (0.02 mmol) in ethanol (0.5 M) was stirred under microwave irradiation at 100°C for 1 h. The resulting precipitate was filtered and purified by column chromatography (dichloromethane / methanol 95:5) to afford the desired compounds CU2, CU3, CU5 and CU6, in 26, 33, 20 and 49% yield, respectively.

[0166] N, 4-Bis(4-chlorophenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine-5-carboxamide (CU2). IR 3404, 3268, 3108, 1707, 1669, 1626, 1510, 1490, 1397, 1320, 1241 , 1089, 1012, 817, 756, 681 , 639, 625 cm’1. HRMS (ESI-TOF (-)) calculated for C18H15CI2N3O2 [M-H] = 374.0540, found 374.0468.

[0167] A / -(4-Chlorophenyl)-4-(4-methoxyphenyl)-6-methyl-2-oxo-1,2,3,4-tetrahydropyrimidine- 5-carboxamide (CU3). IR 3401 , 3271 , 3113, 2929, 1707, 1670, 1628, 1508, 1395, 1322, 1243, 1173, 1088, 1032, 820, 755, 725, 682, 648 cm’1. HRMS (ESI-TOF (+)) calculated for C19H18CIN3O3 [M+H] = 372.1040, found 372.1108. A / -(4-Chlorophenyl)-4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxamide (CU5). IR 3266, 3113, 1711 , 1668, 1593, 1512, 1493, 1467, 1397, 1312, 1245, 1133, 1089, 1029, 1013, 820, 755, 707, 683, 640 cm’1. HRMS (ESI- TOF (+)) calculated for C18H14CI3N3O2 [M+H] = 410.0150, found 410.0238.

[0168] A / -(4-Chlorophenyl)-6-methyl-2-oxo-4-[4-(trifluoromethoxy)phenyl]-1, 2,3,4- tetrahydropyrimidine-5-carboxamide (CU6). IR 3412, 3273, 1708, 1670, 1629, 1507, 1398, 1292, 1244, 1169, 1091 , 821 , 756, 685, 657, 623 cm’1. HRMS (ESI-TOF (+)) calculated for C19H15CIF3N3O3 [M+H] = 426.0750, found 426.0830.

[0169] Compound preparation

[0170] G9a / GSK-3p inhibitors were serially diluted between 5.0 nM and 0.001 nM in 100 % DMSO (Sigma, St. Louis, USA). Then, respective concentrations were subsequently diluted in MilliQ purified water to reach a final concentration ranging between 50 and 0.001 pM in 1 % DMSO in well.

[0171] In vitro determination of G9a (EHMT2) inhibition activity

[0172] The following fluorescent assay was used for determination of the G9a (also called EHMT2, the euchromatic histone methyltransferase 2) inhibition activity (IC50).

[0173] Protocol:

[0174] G9a activity was measured using the G9a Chemiluminescent Assay Kit (Catalog # 52001 L, BPS Bioscience, San Diego, CA, USA), following manufacturer’s instructions. A comparative control compound UNC0638, which corresponds to 2-cyclohexyl- / V-(1-isopropylpiperidin-4-yl)- 6-methoxy-7-(3-(pyrrolidin-1-yl)propoxy)quinazolin-4-amine which is a well-established G9a inhibitor:

[0175] The experiment was run in triplicate, and data are shown in Table 1 below.

[0176] Table 1. ICso G9a of each G9a / GSK3p inhibitor compound.

[0177] In vitro determination of GLP (EHMT1) inhibition activity

[0178] Since G9a and G9a-like (GLP, also known as EHMT1, euchromatin histone lysine methyltransferase 1) are members of the Suv39h subgroup of SET domain-containing molecules, and together they are the key HKMTs for H3K9me1 and H3K9me2. Then, to confirm the selectivity against G9a, we performed the following fluorescent assay for the determination of the GLP inhibition activity (IC50), with substrate and comparative control compound UNC0638, which is a well-established inhibitor with a high potency of G9a and GLP. Unfortunately, the binding to GLP is undesirable in the context of the invention because it gives rise to cytotoxicity.

[0179] Protocol: GLP activity was measured using the GLP Chemiluminescent Assay Kit (Catalog #53007 BPS Bioscience, San Diego, CA, USA). The experiment was run per triplicate and are provided in Table 2 below.

[0180] Table 2. %l nhibition of GLP of each G9a / GSK3p inhibitor compound. Values represented are mean ± Standard error of the mean (SEM);

[0181] Table 2 shows the percentage of inhibition of GLP from different assayed compounds (three different experiments in triplicate). In general, the percentages of inhibition at 10 pM are less than 50% and were lower than compared to the reference compound UNC0638. It is noteworthy that T2, T6, and U4 were the compounds that showed the lowest percentage of inhibition for GLP, suggesting that they were the most selective for G9a.

[0182] In vitro determination of GSK-3 / 3 inhibition activity

[0183] The following fluorescent assay was used for the determination of the GSK-3P inhibition activity (IC50). GSK-3P activity was measured using the GSK-3P Chemiluminescent Assay Kit (Catalog #, 79700 BPS Bioscience, San Diego, CA, USA), following the manufacturer’s instructions.

[0184] Table 3. ICsoGSK3p of each G9a / GSK3p inhibitor compound.

[0185] Parallel Artificial Membrane Permeation Assays - Blood-Brain Barrier (PAMPA-BBB)

[0186] To evaluate the brain penetration of the different compounds, a parallel artificial membrane permeation assay for the blood-brain barrier was used, following the method described by L. Di, et al. “High throughput artificial membrane permeability assay for blood-brain barrier”, Eur. J. Med. Chem. 2003, 38, 223-232. The in vitro permeability (Pe) of fourteen commercial drugs through lipid extract of porcine brain membrane together with the test compounds was determined. Commercial drugs and assayed compounds were tested using a mixture of PBS:EtOH (70:30). Assay validation was made by comparing the experimental permeability with the reported values of the commercial drugs by a bibliography, and linear correlation between the experimental and reported permeability of the fourteen commercial drugs using the parallel artificial membrane permeation assay was evaluated (y=1.623x-1.332; R2=0.9426). Table 4 shows permeability results from the different commercial and assayed compounds (three different experiments in triplicate) and predictive penetration in the CNS.

[0187] Table 4. Permeability in the PAMPA-BBB assay of tested compounds and predictive penetration in the CNS.

[0188] *Spectral absorbance 210-250 nm

[0189] All compounds of the invention showed a markedly improved ability to penetrate the BBB compared to the reference compound UNC0638 (Table 4). EXAMPLE 2. In vivo results in C. elegans

[0190] Statistics analysis

[0191] The statistical analysis was conducted using GraphPad Prism version 9.2 statistical software. Group size may differ depending on power analysis and expertise of the authors. Shapiro-Wilk test to verify data normality for all groups. Data were expressed as the mean ± Standard Error of the Mean (SEM). For normally distributed data, means were compared in One-Way or Two- Way ANOVA analysis of variance (ANOVA), followed by Tukey’s post-hoc analysis. Comparison between groups was also performed by a two-tailed Student’s t-test for independent samples. In contrast, the Mann-Whitney or Kruskal-Wallis test, followed by Dunn’s post-hoc analysis, was used for non-normally distributed data. Statistical significance was considered when p-values were <0.05. T. For behavioral tests, a blinded analysis was performed.

[0192] C. elegans model

[0193] The WT C. elegans strain (N2), the transgenics CL2006 (dvls2 [pCL12(unc-54 / human Ap peptide 1-42 minigene)+rol-6(su1006)]) and BR5706 (byls193 [rab-3p::F3(delta)K280 + myo- 2p::mCherry]; bklslO [aex-3p::hTau V337M + myo-2p::GFP]) provided by the C. elegans Genetic Center were used. Standard methods were used for culturing and observing C. elegans. N2 were propagated at 20°C, while CL2006 and BR5706 worms were maintained at 16°C in a temperature-controlled incubator on a solid nematode growth medium (NGM) seeded with Escherichia coli (E. coli) OP50 (Carolina Biological) strain as a food source.

[0194] Pharmacological treatment and locomotion assay

[0195] Locomotion assay of G9a / GSK-3p inhibitors was assessed to obtain dose-response profile, evaluating the impact of the pharmacology treatment in motor dysfunction presented by the transgenic strains CL2006. Worms were grown with continuous shaking at 180 rpm at 20 °C for 4 days. Each well contained a final volume of 60 pL, comprising 25- 30 animals in the larva 1 (L1) stage diluted in S-medium solution, G9a inhibitor at the appropriate dose (means that the working solution is 2.4-fold more concentrated than the final concentration in the well), and OP50 (Carolina Biological) inactivated by freeze-thaw cycles suspended in S-medium complete solution to a final OD595 of 0.9 measured in the microplate reader. On day 5 of age, worms were transferred from the 96-well plates onto an unseeded NGM plate for 45 minutes before starting the trial, allowing the plates to dry.

[0196] Then, locomotion assays were performed in 30 mm NGM plates, in which the whole surface of the plate was covered by OP50 (Carolina Biological, item: 155073). 5 to 10 adult nematodes were placed in the center of a circle (with 1 cm of diameter) in the seeded 30 mm NGM plates. After 1 min, the number of animals remaining inside the circle was scored as a locomotor defect (“LD”). Motor behavior assays were run in triplicates (n=3), with a total of at least 100 animals tested per compound concentration. Motor index establishes a score where treated animals showing the same motor defect as CL2006 have an index of 0%. By contrast, when animals improve motor behavior comparable to the WT (N2) they get an index of 100%. All the results shown in the figures are calculated using the following formula:

[0197] . . . . . ,n / , (LD CL2006 vehicle - LD CL2006 drug) . > _

[0198] Motor index (%) =1- — x 100

[0199] ' ' (LD CL2006 vehicle - LD WT vehicle)

[0200] LD = locomotion defective Table 5. Values represented are mean ± Standard error of the mean (SEM); n = 3 with at least 90-100 worms CL2006 in each group. Statistical analysis: ordinary One-way ANOVA, post- hoc test Dunnett, *p<0.05, **p<0.01 , ***p<0.001 , ****p<0.0001.

[0201] Table 5 shows that the combination of UNC0638 + Tideglusib reduced the paralysis presented by CL2006. However, it is noteworthy that our compounds showed a better or similar doseresponse profile than the combination of the reference compounds. Thioflavin-S staining AB aggregation

[0202] After 5 days of treatment, adult CL2006 C. elegans were fixed in 4% Paraformaldehyde / Phosphate-buffered saline (PBS) (pH 7.5), for 24 hours at 4°C. Then, worms were permeabilized in 5% fresh [3-mercaptoethanol, 1 % Triton X-100, 125 mm Tris (pH 7.5), at 37°C for another 24 hours. On the last day, nematodes were stained with 0.125% Thioflavin-S (ThS) (Sigma, CAS# 1326-12-1) in 50% ethanol (EtOH) for 2 min, de-stained in 50% EtOH for 2 min, washed 3 times with PBS.

[0203] To prepare the glass slide for microscopy, approximately 10 pL volume was transferred on a droplet of Fluoromount G (Electron Microscopy Sciences, CAT#17984-25). Fluorescence images were acquired using a 20 A~ objective of a fluorescence microscope. AB in the head region of worms were quantified by counting the number of ThS positive spots using Imaged and were expressed as A deposits / anterior area. AB aggregates were scored by an investigator blinded to G9a inhibitor treatments.

[0204] Table 6. Quantification of Thioflavin S-positive particles in the head region of CL2006 strain. Values represented are mean ± Standard error of the mean (SEM); n = 3 with 25 worms in each group. Statistical analysis: Kruskal-Wallis test followed by Dunn’s post-hoc analysis, *p<0.05, **p<0 001, ****p<0.0001 ; Mann-Whitney test,##p<0.01 ,###p<0.001.

[0205] Table 6 shows that the T2 reduced AB aggregation by approximately 56% compared to the vehicle group, being more effective in reducing AB aggregation than well-established UNC0638 and even UNC0638 in combination with Tideglusib, both of which only reduced AB deposition by approximately 37%. EXAMPLE 3. Activity on mouse model of Alzheimer disease

[0206] Statistics analysis

[0207] The statistical analysis was conducted using GraphPad Prism version 9.2 statistical software. Group size may differ depending on power analysis and expertise of the authors. Shapiro-Wilk test to verify data normality for all groups. Data were expressed as the mean ± Standard Error of the Mean (SEM). For normally distributed data, means were compared in One-Way or Two- Way ANOVA analysis of variance, (ANOVA), followed by Tukey’s post-hoc analysis. Comparison between groups was also performed by a two-tailed Student’s t-test for independent samples. In contrast, the Mann-Whitney or Kruskal-Wallis test followed by Dunn’s post-hoc analysis was used for non-normally distributed data. Statistical significance was considered when p-values were <0.05. T. For behavioral tests, a blinded analysis was performed.

[0208] Animal model

[0209] Senescence-accelerated mice prone 8 (SAMP8) is a model of age-related neurodegeneration underlying LOAD. This model, established through phenotypic selection from AKR / J mice, presents AD-like cognitive symptoms and behavioural abnormalities, including anxiety-like behaviour and depression. Additionally, SAMP8 brains are characterized by pathological signatures of AD, including neuroinflammation, synaptic deficits, oxidative stress and aberrant epigenetic dysregulation (C. Grinan-Ferre, et. al., “Understanding Epigenetics in the Neurodegeneration of Alzheimer's Disease: SAMP8 Mouse Model”, J Alzheimers Dis. 2018, 62, 943-963).

[0210] Treatment

[0211] Animals were treated for 15 days with vehicle (control, (20% w / v, (2-hydroxypropyl)-p- cyclodextrin)) or the T2 compound via oral gavage. The test compound was dissolved in 20% w / v (2-hydroxypropyl)-p-cyclodextrin, and the volume of injection was calculated according to the animal weight to reach the precise daily dose. A freshly made weekly replaces the drinking solution. Animals had free access to food and water and were kept under standard temperature conditions (22 ± 2°C) and 12h:12h light-dark cycles (300 lux / 0 lux). After the treatment period, cognitive tests were performed on the animals. After 15 days of daily treatment, mice were studied in the behavioral tests.

[0212] Behavioral tests

[0213] The in vivo models for assessing the efficacy of a test compound in learning and memory impairment were based on Novel Object Recognition Test; NORT, and Object Location Test; OLT. Novel Object Recognition Test (NORT)

[0214] Mice were place 90-degree, two-arm, 25-cm-long, 20-cm-high maze of black polyvinyl chloride. Light intensity in the middle of the field was 30 lux. First, mice were individually habituated to the apparatus for 10 min per day for 3 days. On day 4, the animals were allowed to freely explore two identical objects (A and A or B and B) placed at the end of each arm for a 10 min acquisition trial (first trial-familiarization). Then, a 10-min retention trial (second trial) was carried out 2 h (short-term memory) or 24 h (long-term memory) later. During the Short-term memory retention test, the times that the animal spent exploring the new object (TN) and the old object (TO) was recorded. Twenty-four hours after the acquisition trial, the mice were tested again, with a new object and an object identical to the new one in the previous trial (A and C, or B and C). TN and TO were measured from the video recordings from each trial session. A Discrimination index (DI) was defined as (TN-TO) / (TN+TO). The maze, the surface, and the objects were cleaned with 70% ethanol between the animals’ trials to eliminate olfactory cues.

[0215] The learning and memory paradigm is based on the spontaneous exploratory activity of rodents and does not involve rule learning or reinforcement. The object recognition paradigm has been shown to be sensitive to the effects of aging and cholinergic dysfunction, among others (C. Scali, et al., “Nerve growth factor increases extracellular acetylcholine levels in the parietal cortex and hippocampus of aged rats and restores object recognition”, Neurosci Lett. 1994, 170, 117-20; L. Bartolini, et al., “Aniracetam restores object recognition impaired by age, scopolamine, and nucleus basalis lesions”, Pharmacol. Biochem. Behav. 1996, 53, 277-83). This model has been adapted to mice and validated using pharmacological agents (X. Bengoetxea, et al., “Object recognition test for studying cognitive impairments in animal models of Alzheimer's disease”, Front. Biosci. (Schol Ed). 2015, 7, 10-29).

[0216] Table 7. NORT evaluation by DI after 2h and 24h. Values presented are the mean ± SEM; (SAMP8 Control n = 10, and SAMP8 T2 (5mg / Kg) n = 10); Statistics analysis: Student’s t-test analysis; **p<0.01 ; ****p<0.0001.

[0217] Table 7 above shows working memory results by using Novel object recognition test after G9a inhibition treatment with our candidate T2. Interestingly, we showed a significant improvement in SAMP8 treated with T2 in comparison with SAMP8 Control in short- and long-term memories, suggesting the G9a participation of cognitive impairment in Alzheimer’s Disease.

[0218] Brain tissue dissection

[0219] After behavioral tests, SAMP8 mice were euthanized by cervical dislocation. Brains were immediately removed from the skull. Cortex and hippocampus were then isolated and frozen on powdered dry ice. They were maintained at -80 °C for biochemical experiments. For the Golgi staining protocol, see the procedure in the section “Spine density and Golgi staining protocol”.

[0220] Histones extraction, and Western Blotting of H3K9me2 / H3 total

[0221] Histone extraction was performed following the manufacturer’s instructions using hippocampal tissue from each experimental group (EpiQuik Total Histone Extraction HT Kit, EpiGentek, #GP-0007-192). The samples were resolved in a 14% SDS-gel, as previously described.

[0012] To capture chemiluminescence signals were used Amersham Imager 680 and Western blot quantifications were performed using ImageLab software (Bio-Rad). Immunoblots were probed with anti-H3 K9me2 (1 :1000) (Epigentek, #A-4035), and anti-H3 total signaling, #9715).

[0222] Table 8. Quantification of H3K9me2 in SAMP8 and SAMP8 mice treated with T2. Values presented are mean ± SEM (SAMP8 Control n = 5, and SAMP8 T2 (5mg / Kg) n = 5); Statistics analysis: Student’s t-test analysis, **p<0.01. Table 8 shows the quantification of H3K9me2 a main repressive mark modulated by G9a (X. Chen, et al., “G9a / GLP-dependent histone H3K9me2 patterning during human hematopoietic stem cell lineage commitment”, Genes Dev. 2012, 26, 2499-511). Interestingly, we found a reduction of the repressive histone mark H3K9me2 in the hippocampus of SAMP8 treated with T2 at 5 mg / Kg in comparison with SAMP8 Control.

[0223] Protein extraction, and Western Blotting of p-Tau (Ser 396) and p-Tau (Ser 202, Thr205)

[0224] Brain tissues were homogenized in a cold RIPA lysis buffer supplemented with protease and phosphatase inhibitors, ultrasonicated, and centrifuged (n=5). Firstly, the protein concentration of the supernatants was determined using the Bradford protein assay (Bio-Rad), while later, proteins were separated by SDS-PAGE (12-14%) at 120V and transferred to PVDF membranes by electroblotting at 200 mA for 120 min. Afterward, membranes were blocked in 5% BSA in 0.1 % Tween20-TBS (TBS-T) for 1 hour at room temperature (RT), followed by overnight incubation at 4 °C with the primary antibodies presented (p-Tau (Ser 396) 1 :1000, lnvitrogen / #44-752G; p-Tau (Thr 217) 1 :1000, lnvitrogen / #44-744; Tau Total 1 :1000, lnvitrogen / #AHB0042). By enhancing a chemiluminescence-based detection kit and ChemiDoc XRS+System, we visualized immunoreactive proteins on digital images. Semi- quantitative analyses were carried out using ImageLab Software and results were expressed in Arbitrary Units (AU), considering the control mice group as 100%. Protein loading was routinely monitored and normalized by immunodetection of glyceraldehyde 3-phosphate dehydrogenase (GAPDH).

[0225] Table 9. Quantification of p-Tau (Ser 396), p-Tau (Thr 217) and Tau Total levels in SAMP8 and SAMP8 mice treated with T2. Values presented are mean ± SEM (SAMP8 Control n = 5, and SAMP8 T2 (5mg / Kg) n = 5). Statistics analysis: Student’s t-test analysis, **p<0.05, **p<0.01.

[0226] Table 9 shows that the pharmacological treatment with T2 significantly decreases Tau phosphorylation at both positions (Ser 396, and Thr 217) in SAMP8 compared to the control group.

[0227] Dendritic length, spine density and Golgi staining protocol

[0228] Mice were sacrificed by cervical dislocation and brains were removed from the skull (n=5 whole brain per experimental group). Then, the Golgi staining protocol was developed using the FD Rapid GolgiStain kit according to the manufacturer’s instructions (FD NeuroTechnologies, incs, #PK401). For dendritic branching analysis, images of neurons were collected at 20x magnification in an Olympus BX61 microscope coupled to an Olympus DP70 camera. Measurement of neurite length and complexity was performed using NeuronJ macros and Advanced Sholl Analysis. The number of intersections (branch points) within concentric circles of 10 pm radius was measured and compared between groups. Images for analyzing the spine density were acquired using brightfield microscopy with a 50x oil-objective. All neurites analyzed were around 18 pm and they were at a maximum distance of 150 pm from the soma (J. W. Zhang, et al., “Optimized Golgi-Cox Staining Validated in the Hippocampus of Spared Nerve Injury Mouse Model”, Front Neuroanat. 2020, 14:585513).

[0229] Table 10. Quantification of total dendritic length of intersections and spine density in SAMP8 Control and SAMP8 mice treated with T2. (Values presented are mean ± SEM; (SAMP8 Control n = 100 neurons / 5 mice, and SAMP8 T2 (5mg / Kg) n = 100 neurons / 5); Statistics analysis: Student’s t-test analysis, ****p<0.0001.

[0230] Table 10 shows that the pharmacological treatment with T2 significantly increase the dendritic spine density, and the total dendritic length in SAMP8 in comparison with the control group.

Claims

CLAIMS1 . A compound of formula (I), or a salt, solvate or stereoisomer thereof:wherein:R is one or more radicals independently selected from the group consisting of: H; OH; CN; fluorine; chlorine; (Ci-Cw)haloalkyl; -0-(Ci-Cio)alkyl; (Ci-C )alkyl; -O-(Ci- Cw)haloalkyl; SFs; S(O)2 ’I; NR^R’s; CONR^R’s; and COOR’e;R'i, R'2, R’3, R’4, R’5 and R’e, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (C3-C8)cycloalkyl;X represents O or S;Z represents O or NH; andR1is selected from the group consisting of: a (Ci-Cw)alkyl; or a 4-halophenyl ring; with the proviso that formula (I), or a salt, solvate or stereoisomer thereof, does not contain any one of the following compounds selected from the group consisting of: Ethyl 4-(4- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2- oxo-4-p-tolyl-l,2,3,4-tetrahydropyrimidine-5-carboxylate; Propyl 4-(3-chlorophenyl)-6-methyl- 2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-phenyl-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-m-tolyl-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-methoxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-methoxyphenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-fluorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-fluorophenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(2-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-(3-(trifluoromethoxy)phenyl)- l,2,3,4-tetrahydropyrimidine-5-carboxylate; Isobutyl 4-(3-chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; and Ethyl 4-(3-cyanophenyl)-6-methyl-2-oxo- 1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; for use in preventing or treating an G9a / GSK-3p-mediated disorder in a subject in need thereof, particularly by inhibiting GSK-3P and G9a.

2. The compound for use according to claim 1 , whereinR is selected from the group consisting of: H; fluorine; chlorine; (Ci-Cw)haloalkyl; -O- (Ci-Cio)alkyl; (Ci-C )alkyl and -0-(Gi-Cio)haloalkyl;X represents O or S;Z represents O or NH; andR1a (Ci-C4)alkyl; or a 4-halophenyl ring.

3. The compound for use according to any one of the preceding claims, whereinR is selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; -O- (Ci-C4)alkyl; (Ci-C4)alkyl and -O-(Ci-C4)haloalkyl;X represents O or S;Z represents O or NH; andR1a (Ci-C4)alkyl; or a 4-halophenyl ring.

4. The compound for use according to any one of the preceding claims, wherein the compound is of formula (II), or a salt, solvate or stereoisomer thereof:wherein:R is one or more radicals independently selected from the group consisting of: H; OH;CN; fluorine; chlorine; (Ci-Cw)haloalkyl; -0-(Ci-Cio)alkyl; (Ci-Cw)alkyl; -O-(Ci- Cw)haloalkyl; SFs; S(O)2R’I; NR^R’a; CONR^R’s; and COOR’e;R'i, R'2, R’3, R’4, R’5and R’e, are independently selected from the group consisting of: -H, (Ci-Cw)alkyl; (Ci-Cw)haloalkyl; and (Cs-Csjcycloalkyl; andX represents O or S; with the proviso that formula (II), or a salt, solvate or stereoisomer thereof, does not contain any one of the following compounds selected from the group consisting of: Ethyl 4-(4- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3- chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2- oxo-4-p-tolyl-l,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-phenyl-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-m-tolyl-1 ,2,3,4- tetrahydropynmidme-5-carboxylate; Ethyl 4-(3-methoxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-methoxyphenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-fluorophenyl)-6-methyl-2-oxo-1 ,2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(4-fluorophenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(2-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 4-(3-hydroxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxylate; Ethyl 6-methyl-2-oxo-4-(3-(trifluoromethoxy)phenyl)-1.2.3.4-tetrahydropyrimidine-5-carboxylate; nd Ethyl 4-(3-cyanophenyl)-6-methyl-2-oxo-1.2.3.4-tetrahydropyrimidine-5-carboxylate.

5. The compound for use according to claim 4, whereinR is one or more radicals independently selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; -O-(Ci-C4)alkyl; (Ci-C4)alkyl and -O-(Ci- C4)haloalkyl.

6. The compound for use according to any one of claims 4 or 5, whereinX represents S; andR is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and CF3or CHF2.

7. The compound for use according to claim 6, which is selected from the group consisting of: a) Ethyl 6-methyl-4-phenyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T1); b) Ethyl 4-(4-chlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T2); c) Ethyl 4-(4-methoxyphenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T3); d) Ethyl 6-methyl-2-thioxo-4-(p-tolyl)-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T4); e) Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T5);f) Ethyl 6-methyl-2-thioxo-4-[4-(trifluoromethyl)phenyl]-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T6); g) Ethyl 4-(2,4-dichlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (T7); h) Ethyl 4-(2-chlorophenyl)-6-methyl-2-thioxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxylate (T8); and any salt, solvate or stereoisomer thereof.

8. The compound for use according to any one of claims 4 or 5, whereinX represents O; andR is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and CF3.

9. The compound for use according to claim 8, which is selected from the group consisting of: a) Ethyl 4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U5); b) Ethyl 6-methyl-2-oxo-4-[4-(trifluoromethyl)phenyl]-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U6); c) Ethyl 6-methyl-2-oxo-4-(2,4-dichlorophenyl)-1 ,2,3,4-tetrahydropyrimidine-5- carboxylate (U7); and any salt, solvate or stereoisomer thereof.

10. The compound for use according to any one of claims 1 to 3, wherein the compound is of formula (III), or a salt, solvate or stereoisomer thereof:wherein:R is one or more radicals independently selected from the group consisting of: H; fluorine; chlorine; (Ci-C4)haloalkyl; -O-(Ci-C4)alkyl; (Ci-C4)alkyl; and -O-(Ci- C4)haloalkyl.

11. The compound for use according to the precedent claim, whereinR is one or more radicals independently selected from the group consisting of: H; Cl; - OCH3; CH3and -OCF3.

12. The compound for use according to any one of claims 10 or 11 , which is selected from the group consisting of: a) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-phenyl-1 ,2,3,4-tetrahydropyrimidine-5- carboxamide (CU1); b) A / , 4-Bis(4-chlorophenyl)-6-methyl-2-oxo-1 ,2,3,4-tetrahydropyrimidine-5-carboxamide (CU2); c) / V-(4-Chlorophenyl)-4-(4-methoxyphenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxamide (CU3); d) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-(p-tolyl)-1 ,2,3,4-tetrahydropyrimidine-5- carboxamide (CU4); e) / \ / -(4-Chlorophenyl)-4-(3,4-dichlorophenyl)-6-methyl-2-oxo-1 , 2,3,4- tetrahydropyrimidine-5-carboxamide (CU5); f) / V-(4-Chlorophenyl)-6-methyl-2-oxo-4-[4-(trifluoromethoxy)phenyl]-1 , 2,3,4- tetrahydropyrimidine-5-carboxamide (CU6); and any salt, solvate or stereoisomer thereof.

13. The compound for use according to any one of the precedent claims, wherein the G9a / GSK3B-mediated disorder is a protein-aggregation disease and the compound is for use in the treatment and / or prevention of a protein-aggregation disease.

14. The composition for use according to claim 13, wherein the protein-aggregation disease is Alzheimer’s disease, Parkinson’s disease or Huntington’s disease patients.

15. The compound for use according to any one of claims 1 to 12, wherein the G9a / GSK-3p- mediated disorder is a disease selected from the group consisting of Alzheimer’s disease, amyotrophic lateral sclerosis, age-related tau astrogliopathy, aortic amyloidosis, argyrophilic grain disease, British familial dementia, cardiac amyloidosis, cerebral amyloid angiopathy, chronic traumatic encephalopathy, corneal dystrophies, corticobasal degeneration, Creutzfeldt-Jakob disease, Danish familial dementia, Down syndrome, familial amyloidosis, familial corneal amyloidosis, fatal insomnia, frontotemporal dementia, Gerstmann-Straussler- Scheinker disease, globular glial tauopathy, hereditary cerebral hemorrhage with amyloidosis, Huntington’s disease, inflammation-associated amyloidosis, kuru, Lewy bodies dementia, Mediterranean fever, Niemann-Pick disease type C, Parkinson’s disease, Pick’s disease, primary age-related tauopathy, progressive subcortical gliosis, progressive supranuclear palsysystemic amyloidosis, including transthyretin-associated amyloidosis and light-chain amyloidosis, subacute sclerosing panencephalitis and tuberous sclerosis.

16. The composition for use according to any one of claims 14 or 15, wherein the proteinaggregation disease is Alzheimer’s disease.

Images