FIELD OF THE INVENTION

The present invention is related to the use of indole-dihydro-imidazole derivatives of Formula (I) as a medicament for the treatment of diseases or conditions mediated by Cdk5 and/or GSK3.

BACKGROUND OF THE INVENTION

Cyclin-dependent kinase 5 (Cdk5)

Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine protein kinase with a growing number of identified substrates, such as β-Amyloid precursor protein (β-ΑΡΡ), presenilin-1 , tau protein or neurofilaments (NFs) [Nat. Rev., Mol. Cell Biol., 2001, 2, 749-759. A decade of CDK5. Dhavan R. & Tsai L.H.; Journal of Neurochemistry, 2004, 88, 1313-1326. Cyclin-dependent kinase-5 in neurodegeneration. Shelton S.B. & Johnson G. V. W.; Cell Mol Neurobiol, 2008, 28, 351-369. An Unusual Member of the Cdk Family: Cdk5. Dhariwala F.A. & Rajadhyaksha M.S.]. Cdk5 does not play a role in cell division but plays a pivotal role in the development of the central nervous system (CNS) [Nat. Rev., Mol. Cell Biol., 2001, 2, 749-759. A decade of CDK5. Dhavan R. & Tsai L.H.]. Cdk5 functions beyond neurodevelopment and may play a role in learning and memory, in dopamine signaling in the basal forebrain, in the migration and diferentiation of neurons during histogenesis of the brain, in regulation of the cytoskeletal elements, membrane transport, synaptic function and during neuronal differentiation and axonal extension. Therefore, CDK5 likely plays a role in synaptic plasticity, cellular motility and adhesion, drug addiction and neurodegeneration [Biochimica et Biophysica Acta 1697 (2004) 137- 142. Cdk5, a therapeutic target for Alzheimer's disease?. Tsai L.H., Lee M.S. & Cruz J; Cell Mol Neurobiol, 2008, 28, 351-369. An Unusual Member of the Cdk Family: Cdk5. Dhariwala F.A. & Rajadhyaksha M.S. ; J. Com p. Neurol., 1999, 415, 218-229. Callosal axon guidance defects in p35 (-/-) mice. Kwon Y. T., Tsai L.H. & Crandall J. £.; Journal of Neurochemistry, 2004, 88, 1313-1326. Cyclin-dependent kinase-5 in neurodegeneration. Shelton S.B. & Johnson G. V. W.].

In order for Cdk5 to become active, it must form a heterodimer with p35, p25 (the truncated form of p35), p39 or p29 (the truncated form of p39) [Journal of Neurochemistry, 2004, 88, 1313-1326. Cyclin-dependent kinase-5 in neurodegeneration. Shelton S.B. & Johnson G. V. W.]. Both p35 and p39 have a short half-life in vivo and are subject to ubiquitin-mediated proteosome degradation indicating that Cdk5 activity is normally tightly regulated. Several neurotoxic conditions, including ischemic brain damage, oxidative stress, excitotoxicity and β-amyloid peptide treatment of primary neurons, were shown to induce calpain-mediated p35 cleavage and p25 production. p25, which is neurotoxic, contains all the elements necessary for Cdk5 activation. It efficiently activates Cdk5 both in vitro and in vivo. p25 and p35 have distinct properties: p35 has a very short half-life but p25 is stable and has a much longer half-life; p35 is localized to the membranes but p25 localizes to the cell soma and nucleus. As such, generation of p25 disrupts the normal regulation of Cdk5 by causing prolonged activation and mislocalization of Cdk5 and resulting in hyperphosphorylation of available substrates [Biochimica et Biophysica Acta 1697 (2004) 137- 142. Cdk5, a therapeutic target for Alzheimer's disease?. Tsai L.H., Lee M.S. & Cruz J.].

Although Cdk5 is expressed in most tissues, its activity is found predominantly in post-mitotic neurons [Development, 2003, 119, 1029-1040. Activity and expression pattern of cyclin-dependent kinase 5 in the embryonic mouse nervous system. Tsai L. H. et al.]. The reason for the localized activity of Cdk5 is that its activators, p35 and p39, are expressed almost exclusively in the CNS [Nature, 1994, 371, 423-426. A brain-specific activator of cyclin-dependent kinase 5. Lew et al.; Nature, 1994, 371, 419-423. p35 is a neural-specific regulatory subunit of cyclin-dependent kinase 5. Tsai et al.; FEBS Lett, 1994, 355, 35-40. Precursor of cdk5 activator, the 23 kDa subunit of tau protein kinase II: its sequence and developmental change in brain. Uchida et al.; J. Biol. Chem., 1995, 270, 26897-26903. An isoform of the neuronal cyclin-dependent kinase 5 (Cdk5) activator. Tang et al.; Neuroscience, 2003, 118, 323-334. The cyclin- dependent kinase 5 activator, p39, is expressed in stripes in the mouse cerebellum Jeong et al.; Cell. Mol. Biol. Lett, 2003, 8, 546,547. The role of cyclin-dependent kinase 5 in brain development and degeneration Tsai L.H.]. Although Cdk5 is present predominantly in the nervous system, this enzyme has been implicated to be an important molecule in other systems as well. Since its discovery over a decade ago, an increasing number of the Cdk5 substrates have been reported, making it one of the most versatile kinases. Cdk5 is also known to play an important role in other non- neuronal functions, like myogenesis, haematopoesis, reproduction, prostate cancer, differentiation of lens epithelial and corneal cells, gene-transcription, senescence, insulin secretion, apoptosis, exocytosis/endocytosis, hormonal regulation and wound healing and migration [for a review, see Cell Mol Neurobiol, 2008, 28, 351-369. An Unusual Member of the Cdk Family: Cdk5. Dhariwala F.A. & Rajadhyaksha M.S. ; J. Comp. Neurol., 1999, 415, 218-229. Callosal axon guidance defects in p35 (-/-) mice. Kwon Y. T., Tsai L.H. & Crandall J. £.].

The Cdk5 kinase is activated in cultured neurons following p25 overexpression, and leads to tau hyperphosphorylation and apoptosis. In vitro, certain neurotoxic insults, including exposure to β-amyloid (Αβ), trigger p25 accumulation, Cdk5 activation, and neuronal death, which is averted by pharmacological cdk inhibitors. Moreover, extracellular Αβ deposits induce Cdk5 activation and tau hyperphosphorylation in mutant APP transgenic mice [American Journal of Pathology, 2004, 165, 3, 843-853. Cyclin-Dependent Kinase Inhibitors Attenuate Protein Hyperphosphorylation, Cytoskeletal Lesion Formation, and Motor Defects in Niemann- Pick Type C Mice. Zhang M. et al.; J. Neurosci. Res., 2002, 69, 362-372. p35/ Cdk5 pathway mediates soluble amyloid-beta peptide-induced tau phosphorylation in vitro. Town T. et al.].

Cdk5 has a major activity phosphorylating tau in brain [J. Biochem. (Tokyo),

1997, 121, 179- 188. Physiology and pathology of tau protein kinases in relation to Alzheimer's disease. Imahori K. & Uchida T.]. Cdk5, or downstream kinases, can phosphorylate tau prior to, or independently of, aggregation [Neuron, 2003, 38, 555- 565. Cdk5 Is a Key Factor in Tau Aggregation and Tangle Formation In Vivo. Noble W. et al.]. Many Cdk5 phosphorylation sites on tau have been identified. Ser202, Thr205, Thr212, Ser235, Ser396 and Ser404, all Ser/Thr-Pro sites, are the most likely to be physiologically relevant, as several groups have identified them as Cdk5 sites when tau is phosphorylated in vitro [Journal of Neurochemistry, 2004, 88, 1313-1326. Cyclin- dependent kinase-5 in neurodegeneration. Shelton S.B. & Johnson G. V. W; Neuron, 2003, 38, 555-565. Cdk5 Is a Key Factor in Tau Aggregation and Tangle Formation In Vivo. Noble W. et al.]. Although most of the studies have been carried out in vitro, there is some evidence of tau phosphorylation by Cdk5 in situ and in vivo. Hence, Cdk5 may play a role in the process of NFT formation. Increased Cdk5 activity leads to insoluble aggregation and fibrillization of endogenous tau in the cortex and hippocampus of p25 Tg mice. Furthermore, Cdk5 may indirectly regulate the kinases and phosphatases that act on tau. Cdk5 has been shown to modulate the activity of several protein kinases involved in tau hyperphosphorylation [Neuron, 2003, 40, 471-483. Aberrant Cdk5 Activation by p25 Triggers Pathological Events Leading to Neurodegeneration and Neurofibrillary Tangles. Cruz J.C. et al.; Journal of Neurochemistry, 2004, 88, 1313- 1326. Cyclin-dependent kinase-5 in neurodegeneration. Shelton S.B. & Johnson G. V. W\.

Additionally, it has been demonstrated that Cdk5 inhibitors can reduce β-amyloid (Αβ) level in the mouse brain [J. Neurosci., 2008, 28, 2624- 2632. Interplay between Cyclin-Dependent Kinase 5 and Glycogen Synthase Kinase 3β Mediated by Neuregulin Signaling Leads to Differential Effects on Tau Phosphorylation and Amyloid Precursor Protein Processing. Wen, Y. et al.], since Αβ generation correlates with enhanced Cdk5 activity. Cdk5 has been shown to phosphorylate APP on Thr668 in the cytoplasmic domain. It is reported that T668-phosphorylated APP is up-regulated in AD brain tissues. Moreover, the observations that p25 is elevated in AD brain tissue and in an animal model for β-amyloid and that p25 can be induced by Αβ peptide treatment of primary neurons, indicate that p35 to p25 cleavage may be a downstream event of β- amyloid toxicity [Biochimica et Biophysica Acta 1697 (2004) 137- 142. Cdk5, a therapeutic target for Alzheimer's disease?. Tsai L.H., Lee M.S. & Cruz J.].

Cdk5/p25 might also interact with other pathways such as glycogen synthase kinase 3beta (GSK3beta) and c-JUN kinase. Therefore, Cdk5, and other kinases such as glycogen synthase kinase 3β (Θ8Κ3β), have been identified as prime candidates for AD pathogenesis because they both generate disease associated phospho-epitopes on tau, and they colocalize with filamentous tau aggregates in the brains of patients and in a transgenic mouse model of tauopathy. Cdk5 and GSK3 also regulate Αβ production in vivo [J. Neurosci., 2008, 28, 2624- 2632. Interplay between Cyclin- Dependent Kinase 5 and Glycogen Synthase Kinase 3β Mediated by Neuregulin Signaling Leads to Differential Effects on Tau Phosphorylation and Amyloid Precursor Protein Processing. Wen, Y. et al.].

Pathologies related to Cyclin-dependent kinase 5 (Cdk5)

Deregulation of cyclin-dependent kinases (Cdks), specifically Cdk5, and cytoskeletal protein hyperphosphorylation characterizes a subset of human neurodegenerative diseases, including Alzheimer's disease [Biochimica et Biophysica Acta 1697 (2004) 137- 142. Cdk5, a therapeutic target for Alzheimer's disease?. Tsai L.H., Lee M.S. & Cruz J.], amyotrophic lateral sclerosis [Neurosci. Lett., 1998, 245, 45- 48. Cyclin-dependent kinase-5 is associated with lipofuscin in motor neurones in amyotrophic lateral sclerosis. Bajaj et a/.], Niemann-Pick Type C (NPC) [Neurobiol Dis, 2002, 11, 285-297. Niemann-Pick disease type C yields possible clue for why cerebellar neurons do not form neurofibrillary tangles. Bu B. et a/.], alcohol-induced neurodegeneration [Neurosci. Lett., 2001, 308, 173-176. Ethanol induced changes in cyclin-dependent kinase-5 activity and its activators, P35, P67 (Munc-18) in rat brain Rajgopal Y. & Vemuri M.C.], diffuse Lewy body disease [Am. J. Pathol., 1995, 147, 1465-1476. Cortical and brainstem-type Lewy bodies are immunoreactive for the cyclin-dependent kinase 5. Brion J. P. & Couck A.M.], Parkinson's disease [Proc Natl Acad Sci U S A, 2003, 100, 13650-13655. Cyclin-dependent kinase 5 is a mediator of dopaminergic neuron loss in a mouse model of Parkinson's disease. Smith P.D. et a/.], progressive supranuclear palsy [Neurology, 2002, 58, 589-592. Increase of cdk5 is related to neurofibrillary pathology in progressive supranuclear palsy. Borghi R. et a/.], frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) and other tauopathies [J Biol Chem., 2009, 15, 284(20), 13422-33. Familial FTDP-17 missense mutations inhibit microtubule assembly-promoting activity of tau by increasing phosphorylation at Ser202 in vitro. Han D. et a/.], amnestic mild cognitive impairment [Neurochem Res., 2007, 32(4-5), 655-62. Regional expression of key cell cycle proteins in brain from subjects with amnestic mild cognitive impairment. Sultana R. & Butterfield D.A.], Down syndrome [Neuroscience, 2008, 22, 156(1), 99-106. Abnormal expression of synaptic proteins and neurotrophin-3 in the Down syndrome mouse model Ts65Dn. Pollonini G. et a/.], Huntington disease [J Neurosci., 2008, 1, 28(40), 10090-101. Dopaminergic and glutamatergic signaling crosstalk in Huntington's disease neurodegeneration: the role of p25/cyclin-dependent kinase 5. Paoletti P. et a/.], multiple sclerosis [J Biol Chem., 2004, 31, 279(53), 55833-9. Hyperphosphorylation and aggregation of tau in experimental autoimmune encephalomyelitis. Schneider A. et a/.], and stroke [Proc Natl Acad Sci USA, 2000, 97, 10254-10259. Cyclin-dependent kinases as a therapeutic target for stroke. Osuga H. et al.].

Apart from neurodegenerative diseases, a series of disorders with potential Cdk5 involvement are brain ischemia [Nature Neuroscience, 2003, 6, 1039-1047. Cdk5 activation induces hippocampal CA 1 cell death by directly phosphorylating NMDA receptors. Wang J. et al.], acute neuronal injury [J Neurochem, 2004, 88(6), 1313- 1326. Cyclin-dependent kinase-5 in neurodegeneration. Shelton S.B. & Johnson G. V. W.], cocaine addiction [Nature, 2001, 410, 376-380. Effects of chronic exposure to cocaine are regulated by the neuronal protein Cdk5. Bibb J.A. et al.], electroconvulsive seizures [J. Neurosci., 2000 20, 8965-8971. Induction of Cyclin-Dependent Kinase 5 in the Hippocampus by Chronic Electroconvulsive Seizures: Role of AFosB. Chen J. et al.], fetal alcohol syndrome [Neurosci. Lett, 2001, 308, 173-176. Ethanol induced changes in cyclin-dependent kinase-5 activity and its activators, P35, P67 (Munc-18) in rat brain Rajgopal Y. & Vemuri M.C.], focal cortical dysplasia [Dev Neurosci., 2008, 30(1-3), 96-104. The potential role of cyclin-dependent kinase 5 in focal cortical dysplasia. Sen A. et a/.], hereditary canine spinal muscular atrophy [J. Neuropathol. Exp. Neurol., 1998, 57(11), 991-1099. Alterations in Cyclin-Dependent Protein Kinase 5 (CDK5) Protein Levels, Activity and Immunocytochemistry in Canine Motor Neuron Disease. Green S. et a/.], inclusion body myositis [Neurology 1999, 53, 1671-1676. Aberrant expression of cyclin-dependent kinase 5 in inclusion body myositis Nakano S. et al.], multiple system atrophy [J. Neuropathol. Exp. Neurol., 1998, 57, 690-698. Cyclin-dependent kinase 5 and mitogen-activated protein kinase in glial cytoplasmic inclusions in multiple system atrophy. Nakamura S. et al.], peripheral nerve injury [Neuroreport., 2002, 13, 243-247. Induction of Cdk5 activity in rat skeletal muscle after nerve injury. Fu W. Y. et al.], anxiety and stress [Eur Neuropsychopharmacol., 2010, 20(6), 388-397. Previous stress exposure enhances both anxiety-like behaviour and p35 levels in the basolateral amygdala complex: Modulation by midazolam. Bignante E.A. et al.], epilepsy [Synapse, 2009, 63(11), 1017-1028. Gene expression analysis on anterior temporal neocortex of patients with intractable epilepsy. Xi Z.Q. et al.], prion diseases [J Neurosci., 2009, 8, 29(27), 8743-8751. Phosphorylation of Prion Protein at Serine 43 Induces Prion Protein Conformational Change. Paresa N. et al.], ataxia [Mol Cell Biochem., 2008, 318(1-2), 7-12. Tyrosine hydroxylase expression and Cdk5 kinase activity in ataxic cerebellum. Cheung K.J. et al.], schizophrenia [Neuroreport., 2006, 17(18), 1903-5. Expression of p25 impairs contextual learning but not latent inhibition in mice. Mizuno K. et al.], pain [Cell Cycle, 2006, 5:6, 585-588. Cdk5: A New Player in Pain Signaling. Tej K. et al.], diabetic neuropathy [Rinsho Shinkeigaku., 1999. 39(1), 87-9. New trend in pathogenesis of diabetic neuropathy. Yasuda H.], and diabetes mellitus [Mini Rev Med Chem., 2007, 7(10), 1070-4. Cyclin-dependent kinase 5 (Cdk5): a potential therapeutic target for the treatment of neurodegenerative diseases and diabetes mellitus. Wei F. Y. & Tomizawa K.]. Glycogen synthase kinase-3 (GSK-3)

Glycogen synthase kinase-3 (GSK-3) is a serine/threonine protein kinase comprised of a and β isoforms that are each encoded by distinct genes (Chemistry & Biology, 2000, 7(10), 793-803. Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription. Coghlan et al.; Curr. Opinion Genetics Dev., 2000, 10(5), 508-514. GSK3, a master switch regulating cell- fate specification and tumorigenesis. Kim, L. & Kimmel, A.R.). GSK-3 plays critical roles in development, metabolic homeostasis, neuronal growth and differentiation, cell polarity, cell fate and in modulation of apoptotic potential.

Pathologies related to Glycogen synthase kinase-3 (GSK-3)

Deregulation (usually increase) of GSK-3 activity is believed to play a role in different and important disorders like neurodegenerative disorders [Physiol. Rev., 2004, 84, 361-84. Role of tau protein in both physiological and pathological conditions. Avila, J. et a/.], cardiovascular disease [Circ Res., 2009, 104(11), 1240-52; Role of glycogen synthase kinase-3beta in cardioprotection. Juhaszova M. et al.; Circ J., 2009, 73(7), 1184-92. GSK-3beta, a therapeutic target for cardiomyocyte protection. Miura T. & Miki T], diabetes [Trends. Mol. Med., 2002, 8, 126-32. Glycogen synthase kinase 3: an emerging therapeutic target. Eldar-Finkelman, H.] or viral infections [Virus Res., 2008, 132, 160-73. Residues in human respiratory syncytial virus P protein that are essential for its activity on RNA viral synthesis. Asenjo, A. et al.].

Regarding neurodegenerative disorders and other CNS pathologies, GSK-3 deregulation has been related to Alzheimer's disease [Brain Res Bull., 2009, 80(4-5), 248-50. The role of GSK3 in Alzheimer disease. Hernandez F. et al.], Parkinson's disease [Neuroscience Letters 2009, 449(2), 103-107. Glycogen synthase kinase- 3beta is associated with Parkinson's disease. Masahiro N. & Hideaki H.], frontotemporal dementia [Arch. Neurol., 2008, 65, 1368-74. Association of GSK3B with Alzheimer disease and frontotemporal dementia.. Schaffer, B. et al.], Pick's disease, progressive supranuclear palsy, subacute sclerosing panencephalitis, postencephalitic parkinsonism, dementia pugilistica, guam parkinsonism-dementia complex, corticobasal degeneration, argyrophilic grain disease, familial frontotemporal dementia and parkinsonism linked to chromosome 17 due to mutations in the tau gene (FTDP- 17-tau) and other tauopathies [Brain Research Reviews, 2000, 33, 95 -130. Tau protein isoforms, phosphorylation and role in neurodegenerative disorders. Buee, L. et al.], AIDS associated dementia [J Neuroimmune Pharmacol., 2007, 2(1), 93-96. Glycogen synthase kinase 3 beta (GSK-3 beta) as a therapeutic target in neuroAIDS. Dewhurst S. et al.], Huntington's disease [J Biol Chem., 2002, 277(37), 33791-8. Glycogen synthase kinase-3beta inhibitors prevent cellular polyglutamine toxicity caused by the Huntington's disease mutation. Carmichael J. et al.], Lewy body disease [Neuropathology, 2003, 23(3), 199-202. Glycogen synthase kinase-3beta phosphorylates synphilin-1 in vitro. Tanji K. et al.], bipolar disorder [Neurosci Biobehav Rev., 2007, 31(6), 920-931; GSK-3 is a viable potential target for therapeutic intervention in bipolar disorder. Roew M.K. et al.; Bipolar Disord., 2002, 4(2), 137-144. Glycogen Synthase Kinase-3fi, mood stabilizers, and neuroprotection. Li X. et al.], depression [J Pharmacol Sci., 2009, 110(1), 14-28. Lithium and neuropsychiatric therapeutics: neuroplasticity via glycogen synthase kinase-3beta, beta-catenin, and neurotrophin cascades. Wada A.], schizophrenia [Drug News Perspect., 2007, 20(7), 437-45. The role of glycogen synthase kinase-3beta in schizophrenia. Koros E. & Dorner-Ciossek C; Trends Neurosci., 2007, 30(4), 142-9. Schizophrenia as a GSK-3 dysregulation disorder. Lovestone S. et al.], epilepsy [J Neurochem., 1999, 72(3), 1327-30. The mood-stabilizing agent valproate inhibits the activity of glycogen synthase kinase-3. Chen G. et al.], mood disorders [Curr Drug Targets., 2006, 7(11), 1421-34. Glycogen synthase kinase-3 (GSK3) in psychiatric diseases and therapeutic interventions. Jope R.S. & Roh M.S.], autism [Proc Natl Acad Sci U S A., 2008, 105(4), 1333-8. Role of GSK3 beta in behavioral abnormalities induced by serotonin deficiency. Beaulieu J.M. et al.], attention deficit hyperactivity disorder [Proc Natl Acad Sci U S A., 2004, 101(14), 5099-104. Lithium antagonizes dopamine-dependent behaviors mediated by an AKT/glycogen synthase kinase 3 signaling cascade. Beaulieu J.M. et al.], Down's syndrome [FASEB J., 2008, 22(9), 3224-33. Overexpression of DyrkIA contributes to neurofibrillary degeneration in Down syndrome. Liu F. et al.], diseases associated with ischemia/reperfusion and shock [Shock., 2007, 27(2), 113-23. Glycogen synthase kinase 3beta as a target for the therapy of shock and inflammation. Dugo L. et al.], brain injury [Neurol Res., 2001, 23(6), 588-92. Different expression of glycogen synthase kinase-3beta between young and old rat brains after transient middle cerebral artery occlusion. Sasaki C. et al.], multiple sclerosis [Trends Immunol., 2010, 31(1), 24-31. Innate and adaptive immune responses regulated by glycogen synthase kinase-3 (GSK3). Beurel E. et al.] and other autoimmune and inflammatory diseases afflicting the CNS [J. Immunol., 2008, 181(1), 338-45. Lithium prevents and ameliorates experimental autoimmune encephalomyelitis. De Sarno P. et al.], spinocerebellar ataxia type 1 [PLoS Med, 2007, 4(5), 836-847. Lithium therapy improves neurological function and hippocampal dendritic arborization in a spinocerebellar ataxia type 1 mouse model. Watase K. et al.], cerebral bleeding for example, due to solitary cerebral amyloid angiopathy [Neuroscience., 2008, 153(2), 414-27. Accumulation of beta-amyloid in the brain microvessels accompanies increased hyperphosphorylated tau proteins following microsphere embolism in aged rats. Han F. et a/.] or amyotrophic lateral sclerosis [Brain Res., 2008, 1196, 131-139. Upregulation of GS 3/3 expression in frontal and temporal cortex in ALS with cognitive impairment (ALSci). Yang W. et a/.].

In addition to its possible relevance to prevent neurodegeneration, GSK3 inhibitors may also be useful to foster other forms of neuronal repair, including axon regeneration [J. Neurosci., 2008, 28, 8914-28. Inactivation of glycogen synthase kinase 3 promotes axonal growth and recovery in the CNS. Dill, J. et a/.].

During the last few years, GSK-3 has been identified as a regulator of many components of the immune system, suggesting it might be a plausible therapeutic target in inflammatory and autoimmune diseases, such as chronic inflammatory diseases including rheumatoid arthritis, inflammatory bowel disease and psoriasis [Eur J Biochem., 2001, 268(19), 5001-10. The role of protein phosphorylation in human health and disease. Cohen P.], arthritis [Clin. Immunol., 2006, 120, 57-67. Glycogen synthase kinase-3b inhibition attenuates the degree of arthritis caused by type II collagen in the mouse. Cuzzocrea, S. et al.], peritonitis [Immunity, 2006, 24, 563-574. IFN-g suppresses IL-10 production and synergizes with TLR2 by regulating GSK3 and CREB/AP-1 proteins. Hu, X. et a/.], systemic inflammation, renal dysfunction and hepatotoxicity in endotoxemia [Crit. Care Med., 2005, 33, 1903-1912. GSK-3b inhibitors attenuate the organ injury/dysfunction caused by endotoxemia in the rat. Dugo, L. et a/.], asthma [Am J Physiol Lung Cell Mol Physiol., 2009, 296(2), L176-84. Airway smooth muscle hyperplasia and hypertrophy correlate with glycogen synthase kinase-3(beta) phosphorylation in a mouse model of asthma. Bentley J.K. et al.], sepsis [J. Biochem. Cell. Biol., 2005, 37, 2226-2238. GSK-3b inhibitors reduce protein degradation in muscles from septic rats and in dexamethasone treated myotubes. Int. Evenson, A.R. et a/.], colitis [Br. J. Pharmacol., 2006, 147, 575-582. Reduction of experimental colitis in the rat by inhibitors of glycogen synthase kinase-3b. Whittle, B.J. et a/.], inflammation-induced organ injury caused by hemorrhage and resuscitation [Shock, 2006, 25, 485-491. Glycogen synthase kinase-3b inhibitors protect against the organ injury and dysfunction caused by hemorrhage and resuscitation. Dugo, L. et a/.], inflammatory injury in chronic renal allograft disease [Am J Transplant, 2008, 8(9), 1852-63. Glycogen synthase kinase 3beta: a novel marker and modulator of inflammatory injury in chronic renal allograft disease. Gong R. et a/.] or lupus [Int. J. Immunopharmacol., 1995, 17, 581-592. Lithium chloride enhances survival of NZBAA/ lupus mice: influence of melatonin and timing of treatment. Lenz, S.P. et a/.]. Among cardiovascular disorders related to GSK-3 are heart disease [Circ. Res., 2002, 90, 1055-63. Glycogen synthase kinase-3beta: a novel regulator of cardiac hypertrophy and development. Hardt, S.E. & Sadoshima, J.], atherosclerosis [Am J Pathol., 2009, 174(1), 330-42. Valproate attenuates accelerated atherosclerosis in hyperglycemic apoE-deficient mice: evidence in support of a role for endoplasmic reticulum stress and glycogen synthase kinase-3 in lesion development and hepatic steatosis. Bowes A.J. et al.], hypertension [J. Clin. Invest, 2002, 109(3), 373-381. Fas receptor signaling inhibits glycogen synthase kinase 3β and induces cardiac hypertrophy following pressure overload. Badorff C. et a/.], restenosis [Cardiovasc Res., 2010, Epub. Delayed Re-endothelialization with Rapamycin-coated Stents is Rescued by the Addition of a Glycogen Synthase Kinase 3 Beta Inhibitor. Ma X. et a/.] or leukopenia [Gallicchio, V. S. (1991) in Lithium and the Cell, ed. Birch, N. J. (Academic, San Diego), pp. 185-198.].

Additional pathologies associated with GSK-3 are metabolic syndrome X [Curr Pharm Des., 2004, 10(10), 1105-37. Discovery and development of GSK3 inhibitors for the treatment of type 2 diabetes. Wagman A.S. et a/.], hair loss [J Clin Invest, 2010, 120(2), 446-56. Neural Wiskott-Aldrich syndrome protein modulates Wnt signaling and is required for hair follicle cycling in mice. Lyubimova A. et a/.], severe acute respiratory syndrome coronavirus [J Biol Chem., 2009, 284(8), 5229-39. Glycogen synthase kinase-3 regulates the phosphorylation of severe acute respiratory syndrome coronavirus nucleocapsid protein and viral replication. Wu C.H. et a/.], cocaine addiction [J Neurochem., 2009, 111(6), 1357-68. Glycogen synthase kinase 3beta in the nucleus accumbens core mediates cocaine-induced behavioral sensitization. Xu CM. et a/.], bone loss [Life Sci., 2009, 85(19-20), 685-92. Inhibition of glycogen synthase kinase-3beta attenuates glucocorticoid-induced bone loss. Wang F.S. et a/.], fragile X syndrome (FXS) [Biochem Pharmacol., 2010, 79(4), 632-46. Lithium ameliorates altered glycogen synthase kinase-3 and behavior in a mouse model of fragile X syndrome. Yuskaitis C.J. et a/.] or glaucoma [J Clin Invest, 2008, 118(3), 1056-64. Increased expression of the WNT antagonist sFRP-1 in glaucoma elevates intraocular pressure. Wang W.H. et al.].

GSK-3 inhibitors

For a further review of GSK-3 inhibitors and their use as potential treatments for these pathologies, please reference to Nature Reviews, 2004, 3, 479-487. GSK3 inhibitors: development end therapeutic potential. Cohen, P. & Goedert, M.; Mini- Reviews in Medicinal Chemistry, 2009, 9(9), 1024-1029. GSK3 Inhibitors and Disease. Hernandez, F. et al.; Curr. Opin. Drug Discov. Develop., 2008, 11(4), 533-543. Glycogen synthase kinase-3 (GSK-3) inhibitors reach the clinic. Medina, M. & Castro, A.; John Wiley & Sons, Inc., 2006. Glycogen Synthase Kinase 3 (GSK-3) and its inhibitors. Chapter 14. Eds: Martinez, A., Castro, A. & Medina, M.

Several GSK-3 inhibitors like indirubines [J. Biol. Chem., 2001, 276, 251-60. Indirubins inhibit glycogen synthase kinase-3 beta and CDK5/p25, two protein kinases involved in abnormal tau phosphorylation in Alzheimer's disease. A property common to most cyclin-dependent kinase inhibitors?. Leclerc, S. et al.], maleimides [Bioorg. Med. Chem. Lett, 2001, 11, 635-9. 3-Anilino- 4-arylmaleimides: potent and selective inhibitors of glycogen synthase kinase-3 (GSK-3). Smith, D. et al.], 3-amino pyrazoles [Bioorg. Med. Chem. Lett., 2003, 13, 1581-4. 5-arylpyrazolo[3,4-b]pyridazines: potent inhibitors of glycogen synthase kinase-3 (GSK-3). Witherington, J. et al.], paullones [Eur. J. Biochem., 2000, 267, 5983-94. Paullones are potent inhibitors of glycogen synthase kinase-3beta and cyclin-dependent kinase 5/p25. Leost, M. et al.], thiazoles [J. Biol. Chem., 2003, 278, 45937-45. Structural insights and biological effects of glycogen synthase kinase 3-specific inhibitor AR-A014418. Bhat, R. et al.] or thiadiazolidinones [J. Med. Chem., 2002, 45, 1292-9. First non-ATP competitive glycogen synthase kinase 3 beta (GSK-3beta) inhibitors: thiadiazolidinones (TDZD) as potential drugs for the treatment of Alzheimer's disease. Martinez, A. et al.] have been described.

In view of the above, it is of great potential therapeutic interest to find further GSK-3 inhibitors, which may be useful for treating several diseases or conditions as detailed above.

Indole-dihydro-imidazole derivatives

Some indole-dihydro-imidazole derivatives have been described in the literature, for example in Org. Biomol. Chem., 2008, 6, 2765-2771 , wherein some particular compounds have been described having cytotoxic properties.

BRIEF DESCRIPTION OF THE INVENTION

It has been found that indole-dihydro-imidazole derivatives of Formula (I):

(I) surprisingly inhibit Cdk5 and GSK3 activity, thus having a potential therapeutic interest for the treatment of diseases or conditions wherein Cdk5 and GSK3 are involved and/or a modulation thereof is necessary.

R ¹ , R ² , R ³ and R ⁴ in Formula I have the following meanings:

R ¹ is selected from methyl and hydrogen or is absent, depending on the position of the double bond in the imidazole ring;

R ² is selected from hydroxyl and hydrogen;

R ³ is selected from bromo and hydrogen;

R ⁴ is selected from hydroxyl and hydrogen.

Therefore, according to a first aspect, the present invention is directed to a compound of Formula (I), as defined above, or a tautomer, salt, solvate or prodrug thereof, for its use in the treatment of a disease or condition mediated by Cdk5 or GSK3.

According to a further aspect, the present invention is directed to the use of a compound of Formula (I), as defined above, or a tautomer, salt, solvate or prodrug thereof, for the manufacture of a medicament for the treatment of a disease or condition mediated by Cdk5 or GSK3.

According to another aspect, the present invention is directed to a method of treatment of a disease or condition mediated by Cdk5 or GSK3, said method comprising administering a compound of Formula (I), as defined above, or a tautomer, salt, solvate or prodrug thereof, to a patient in need of treatment.

Finally, another aspect of the present invention relates to the use of a compound of formula (I), or a tautomer, salt, solvate or prodrug thereof, as defined above, as a reactive for a biological assay. DETAILED DESCRIPTION OF THE INVENTION

The disease or condition mediated by Cdk5 or GSK3 in the frame of the present invention is preferably selected from neurodegenerative disorders, CNS pathologies, inflammatory and autoimmune diseases, cardiovascular diseases, diabetes mellitus and viral infections.

Particularly, the neurodegenerative disorder or CNS pathology is preferably, but not limited to, selected from Alzheimer's disease, Parkinson's disease, tauopathies, frontotemporal dementia, Pick's disease, Niemann-Pick Type C (NPC), progressive supranuclear palsy, subacute sclerosing panencephalitis, postencephalitic parkinsonism, dementia pugilistica, guam parkinsonism-dementia complex, corticobasal degeneration, argyrophilic grain disease, familial frontotemporal dementia and parkinsonism linked to chromosome 17 due to mutations in the tau gene (FTDP- 17-tau), AIDS associated dementia, Huntington's disease, Lewy body disease, bipolar disorder, depression, schizophrenia, epilepsy, mood disorders, autism, attention deficit hyperactivity disorder, Down's syndrome, brain ischemia and diseases associated with ischemia/reperfusion and shock, brain injury, multiple sclerosis and other autoimmune and inflammatory diseases afflicting the CNS, spinocerebellar ataxia type 1 , cerebral bleeding, for example due to solitary cerebral amyloid angiopathy, amyotrophic lateral sclerosis, alcohol-induced neurodegeneration, amnestic mild cognitive impairment, stroke, fragile X syndrome (FXS), acute neuronal injury, electroconvulsive seizures, focal cortical dysplasia, hereditary canine spinal muscular atrophy, multiple system atrophy, anxiety and stress, prion diseases, ataxia or pain.

The inflammatory and autoimmune disease is preferably selected from chronic inflammatory diseases including rheumatoid arthritis, inflammatory bowel disease and psoriasis, arthritis, peritonitis, systemic inflammation, renal dysfunction and hepatotoxicity in endotoxemia, asthma, sepsis, colitis, inflammation-induced organ injury caused by hemorrhage and resuscitation, inflammatory injury in chronic renal allograft disease or lupus.

The cardiovascular disorder is preferably selected from heart disease, brain ischemia, atherosclerosis, hypertension, restenosis or leukopenia.

The Cdk5 and GSK3 mediated disease may also be selected from additional pathologies, such as metabolic syndrome X, hair loss, severe acute respiratory syndrome coronavirus, cocaine addiction, bone loss, glaucoma, fetal alcohol syndrome, inclusion body myositis, peripheral nerve injury and diabetic neuropathy. Therefore, the Cdk5 and GSK3 mediated disease may be selected from Alzheimer's disease, Parkinson's disease, tauopathies, frontotemporal dementia, Pick's disease, Niemann-Pick Type C (NPC), progressive supranuclear palsy, subacute sclerosing panencephalitis, postencephalitic parkinsonism, dementia pugilistica, guam parkinsonism-dementia complex, corticobasal degeneration, argyrophilic grain disease, familial frontotemporal dementia and parkinsonism linked to chromosome 17 due to mutations in the tau gene (FTDP-17-tau), AIDS associated dementia, Huntington's disease, Lewy body disease, bipolar disorder, depression, schizophrenia, epilepsy, mood disorders, autism, attention deficit hyperactivity disorder, Down's syndrome, diseases associated with ischemia/reperfusion and shock, brain injury, multiple sclerosis and other autoimmune and inflammatory diseases afflicting the CNS, spinocerebellar ataxia type 1 , cerebral bleeding, for example due to solitary cerebral amyloid angiopathy, amyotrophic lateral sclerosis, alcohol-induced neurodegeneration, amnestic mild cognitive impairment, stroke, chronic inflammatory diseases including rheumatoid arthritis, inflammatory bowel disease and psoriasis, arthritis, peritonitis, systemic inflammation, renal dysfunction and hepatotoxicity in endotoxemia, asthma, sepsis, colitis, inflammation-induced organ injury caused by hemorrhage and resuscitation, inflammatory injury in chronic renal allograft disease, lupus, heart disease, brain ischemia, atherosclerosis, hypertension, restenosis or leukopenia, metabolic syndrome X, hair loss, severe acute respiratory syndrome coronavirus, cocaine addiction, bone loss, fragile X syndrome (FXS), glaucoma, acute neuronal injury, electroconvulsive seizures, fetal alcohol syndrome, focal cortical dysplasia, hereditary canine spinal muscular atrophy, inclusion body myositis, multiple system atrophy, peripheral nerve injury, anxiety and stress, prion diseases, ataxia, pain and diabetic neuropathy.

The compounds of Formula I may be in the form of tautomers, in the form of salts, preferably pharmaceutically acceptable salts, in the form of solvates or in the form of prodrugs.

The term "pharmaceutically acceptable salts" refers to salts which, upon administration to the recipient are capable of providing (directly or indirectly) a compound as described herein. The preparation of salts can be carried out by methods known in the art. Preferably, "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a human. Preferably, as used herein, the term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

For instance, pharmaceutically acceptable salts of compounds provided herein are synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts are, for example, prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of the two. Generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. Examples of the acid addition salts include mineral acid addition salts such as, for example, hydrochloride, hydrobromide, hydroiodide, sulphate, nitrate, phosphate, and organic acid addition salts such as, for example, acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methanesulfonate and p-toluenesulfonate. Examples of the alkali addition salts include inorganic salts such as, for example, sodium, potassium, calcium, ammonium, magnesium, aluminium and lithium salts, and organic alkali salts such as, for example, ethylenediamine, ethanolamine, Ν,Ν-dialkylenethanolamine, triethanolamine, glucamine and basic aminoacids salts.

The term "prodrug" as used in this application is defined here as meaning a chemical compound having undergone a chemical derivation such as substitution or addition of a further chemical group to change (for pharmaceutical use) any of its physico-chemical properties, such as solubility or bioavailability, e.g. ester and ether derivatives of an active compound that yield the active compound per se after administration to a subject. Examples of well known methods of producing a prodrug of a given acting compound are known to those skilled in the art and can be found e.g. in Krogsgaard-Larsen et al., Textbook of Drug design and Discovery, Taylor & Francis (April 2002).

Particularly favoured prodrugs are those that increase the bioavailability of the compounds of this invention when such compounds are administered to a patient (e.g., by allowing an orally administered compound to be more readily absorbed into the blood) or which enhance delivery of the parent compound to a biological compartment (e.g., the brain or lymphatic system) relative to the parent species.

The term "solvate" according to this invention is to be understood as meaning any form of the active compound according to the invention which has another molecule (most likely a polar solvent) attached to it via a non-covalent bonding. Examples of such solvates include hydrates and alcoholates, e.g. methanolates.

The preparation of salts, solvates and prodrugs can be carried out by methods known in the art. It will be appreciated that non-pharmaceutically acceptable salts, solvates or prodrugs also fall within the scope of the invention since those may be useful in the preparation of pharmaceutically acceptable salts, solvates or prodrugs.

The compounds of the present invention may exhibit tautomerism. Tautomers are one of two or more structural isomers of a compound that exist in equilibrium and are readily converted from one isomeric form to another. Common tautomeric pairs are amine-imine, amide-imidic acid, keto-enol, lactam-lactim, etc. For example, the compounds of the present invention may be equally represented as:

Generally a "therapeutically effective amount" of the compound of the invention or a pharmaceutical composition thereof will depend on the relative efficacy of the compound chosen, the severity of the disorder being treated and the weight of the sufferer. However, active compounds will typically be administered once or more times a day for example 1 , 2, 3 or 4 times daily, with typical total daily doses in the range of from 0.1 to 1000 mg/kg/day.

The term "treatment" or "to treat" in the context of this specification means administration of a compound or formulation according to the invention to prevent, ameliorate or eliminate the disease or one or more symptoms associated with said disease. "Treatment" also encompasses preventing, ameliorating or eliminating the physiological sequelae of the disease.

The term "ameliorate" in the context of this invention is understood as meaning any improvement on the situation of the patient treated - either subjectively (feeling of or on the patient) or objectively (measured parameters).

Preferably, the compounds of Formula I are selected from trachycladindoles A-G, for which in Formula (I) R ¹ to R ⁴ have the following meanings: Trachycladindole R R R R

A Br H H H

B Br H H Me

C Br OH H H

D Br OH H Me

E Br H OH Me

F Br OH OH Me

G H H H H or a tautomer, salt, solvate or prodrug thereof.

An additional aspect of the present invention relates to the use of a compound of formula (I), or a tautomer, salt, solvate or prodrug thereof, as defined above, as a reactive for a biological assay. In a preferred embodiment, the biological assay consists in the Cdk5 or GSK3 inhibition. More preferably, the biological assay is an in vitro assay.

EXAMPLES

OBTENTION AND IDENTIFICATION

The isolation, characterization and structure elucidation of trachycladindoles A-G were described for the first time by Capon R.J., Peng C. & Dooms C. (Trachycladindoles A-G: cytotoxic heterocycles from an Australian marine sponge, Trachycladus laevispirulifer. Organic & Biomolecular Chemistry, 2008, 6, 2765-2771 ). This document is herewith incorporated by reference into the present application.

BIOLOGICAL DATA

Example 1 : Cdk5 assay

The enzymatic activity of Cdk5 was determined with a commercial system based on the Z'-LYTE ^® technology, available from Life Technologies (Carlsbad, CA, USA), using human recombinant Cdk5/p35 heterodimer, also available from Invitrogen as the enzyme source. This technology utilizes the fluorescence resonance energy transfer ("FRET") process between fluorescein and coumarin. The assay principle is based on the differential sensitivity of phosphorylated and non-phosphorylated peptide to proteolytic cleavage, which precludes the energy transfer process between the two fluorophores attached to both sides of the cleavage site. Hence, phosphorylation by Cdk5/p35 will yield a phosphopeptide which cannot be hydrolyzed by a suitable protease and energy transfer between the two fluorophores will occur. Opposingly, lack of phosphorylation will cause peptide hydrolysis hence lack of energy transfer. The assay is performed in 384-well black plates, in a final volume of 10 μΙ, with 10 nM enzyme concentration, in 50 mM Hepes pH 7.5, 50 mM MgCI2, 5 mM EGTA and 0.05% Brij-35, using 12.5 μΜ ATP and 2 μΜ substrate peptide. The latter is a synthetic peptide provided by Invitrogen, under the commercial name "Ser/Thr 12 peptide", and it is based upon the sequence of Cdk7, containing Ser-164 and a (SerThr)-Pro-X- (Lys'Arg) consensus sequence, wherein X is any residue. The peptide is labeled at both ends with fluorescein and coumarin. The assay is carried out in the presence of different concentrations of the tested compound, at a final 1 % (v/v) DMSO concentration. After a 60 min incubation, at room temperature, 5 μΙ of a commercial protease solution (sold by the same vendor in the assay kit) is added and a subsequent 1 h incubation at room temperature is performed, before adding 5 μΙ of a suitable "stop solution", also provided by the vendor in the kit. After that, fluorescence intensity is recorded, monitoring emission at both 445 and 520 nm, upon excitation at 400 nm. An emission ratio is finally calculated, using the quotient among the emission at 445 nm divided by that at 520 nm.

In the assay plate, several wells are included as a control for full enzyme activity, these wells do not contain any inhibitor or tested substance. Likewise, several wells are also included as a control for lack of enzyme activity, thus these wells do not contain inhibitor nor enzyme. The emission ratio of each tested sample is normalized to that of the control wells, so that for every compound concentration the percentage of inhibition is calculated by using the following equation:

% Inhibition = 100 · ^{E ~ S}

E-B where "S" is the emission from the wells with the tested sample, "E" is the average emission from control wells with full enzyme activity and "B" is the average emission from wells with full enzyme inhibition. Inhibition values obtained for every compound concentration are finally used to calculate the pEC50 of the tested compound, this parameter being the negative value of the logarithm of the compound concentration, in M units, causing 50% of its maximum effect (i.e., the higher the pEC50 value the higher the potency of the compound). For that purpose the data were fitted to the following equation using the nonlinear regression function of GraphPad™ Prism 5.0 (GraphPad Software Inc.):

where "L" is the lower asymptote of the theoretical sigmoidal curve, "H" is the higher asymptote, "C" is the concentration of compound in M units and "N" is the Hill coefficient.

In Table 1 , the pEC50 values obtained for some compounds of Formula I are indicated:

Table 1

Example 2: GSK-3 assay

The enzymatic activity of GSK3P was determined with a commercial system based on the Z'-LYTE ^® technology, available from Life Technologies (Carlsbad, CA, USA), using human recombinant GSK3P (N-terminal 6His-tagged recombinant enzyme with an H350L mutation) from Millipore (Billerica, MA, USA) as the enzyme source. This technology utilizes the fluorescence resonance energy transfer ("FRET") process between fluorescein and coumarin. The assay principle is based on the differential sensitivity of phosphorylated and non-phosphorylated peptide to proteolytic cleavage, which precludes the energy transfer process between the two fluorophores attached to both sides of the cleavage site. Hence, phosphorylation by GSK3P will yield a phosphopeptide, which cannot be hydrolyzed by a suitable protease and energy transfer between the two fluorophores will occur. Opposingly, lack of phosphorylation will cause peptide hydrolysis hence lack of energy transfer. The assay is performed in 384-well black plates, in a final volume of 10 μΙ, with 2 nM enzyme concentration in 50 mM Hepes pH 7.5, 50 mM MgCI ₂ , 5 mM EGTA and 0.05% Brij-35, using 12.5 μΜ ATP and 2 μΜ substrate peptide. The latter is a synthetic peptide, provided by Invitrogen under the commercial name "Ser/Thr 9 peptide", and it is based upon the sequence of a GSK3P substrate protein (glycogen synthase I) containing Ser-641 . The peptide is labeled at both ends with fluorescein and coumarin. The assay is carried out in the presence of different concentrations of the tested compound, at a final 1 % (v/v) DMSO concentration. After a 60 min incubation at room temperature, 5 μΙ of a commercial protease solution (sold by the same vendor in the assay kit) is added and a subsequent 1 h incubation at room temperature is performed, before adding 5 μΙ of a suitable "stop solution", also provided by the vendor in the kit. After that, fluorescence intensity is recorded, monitoring emission at both 445 and 520 nm, upon excitation at 400 nm. An emission ratio is finally calculated, using the quotient among the emission at 445 nm divided by that at 520 nm.

The emission ratios for each concentration of compound are finally used to calculate the pEC50 value, according to the procedure described above for Cdk5.

In Table 2, the pEC50 values obtained for some compounds of Formula I are indicated:

Table 2

GSK-3 inhibition

Compound No Structure

pEC50 average

Compound 1 See table 1 4.45

Compound 2 See table 1 4.13

Compound 3 See table 1 4.95

Compound 4 See table 1 4.81

Compound 5 See table 1 4.01

Compound 6 See table 1 5.08

Compound 7 See table 1 4.37