BR122020008941B1 - COMBINATIONS COMPRISING POSITIVE ALLOSTERIC MODULATORS OR ORTHOSTERIC AGONISTS OF THE METABOTROPIC GLUTAMATERGIC RECEPTOR SUBTYPE 2, PHARMACEUTICAL COMPOSITION, AND THEIR USES - Google Patents

Abstract

The present invention relates to combinations comprising a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype orthosteric agonist compound. 2 or a pharmaceutically acceptable salt thereof, and a synaptic vesicle protein 2A ("SV2A") ligand.

Description

Divided from BR112016016390-7, filed on 01.20.2015. AREA OF INVENTION

[001] The present invention relates to combinations comprising a positive allosteric modulator ("PAM") of the metabotropic glutamatergic receptor subtype 2 ("mGluR2") or a pharmaceutically acceptable salt or solvate thereof, or a compound metabotropic glutamatergic receptor subtype 2 orthosteric agonist or a pharmaceutically acceptable salt or solvate thereof, and a synaptic vesicle protein 2A ("SV2A") ligand.

BACKGROUND OF THE INVENTION

[002] Epilepsy describes a condition in which a person has recurrent seizures due to an underlying, chronic process. Epilepsy refers to a chronic phenomenon rather than a single disease entity, as there are many forms and causes of epilepsy. Using a definition of epilepsy as two or more unprovoked seizures, the incidence of epilepsy is estimated to be approximately 0.3 to 0.5 percent in different populations globally, with the prevalence of epilepsy estimated to be 5 to 10 persons per 1000.

[003] An essential step in evaluating and managing a patient with a seizure is to determine the type of seizure that has occurred. The main characteristic that distinguishes the different categories of seizures is whether the seizure activity is partial (synonymous with focal) or generalized.

[004] Partial seizures are those in which the seizure activity is restricted to discrete areas of the cerebral cortex. If consciousness is fully retained during the seizure, the clinical manifestations are considered to be relatively simple and the seizure is termed a simple-partial seizure. If consciousness is affected, the seizure is called a partial-complex seizure. An additional important subgroup comprises those seizures that begin as partial seizures and then spread diffusely throughout the cortex, which are known as partial seizures with secondary generalization.

[005] Generalized seizures involve diffuse regions of the brain simultaneously in a bilaterally symmetrical fashion. Absence or petit mal seizures are characterized by brief, sudden lapses of consciousness without loss of postural control. Atypical absence seizures typically include a longer duration of lapse in consciousness, less abrupt onset and cessation, and more obvious motor signs that may include focal or lateralizing features. Tonic-clonic or grand mal seizures, the main type of generalized seizures, are characterized by an abrupt onset without warning. The initial phase of the seizure is usually tonic contraction of muscles, impaired breathing, a marked increase in sympathetic tone leading to increased heart rate, blood pressure, and pupil size. After 10-20 s, the tonic phase of the seizure typically evolves into the clonic phase, produced by overlapping periods of muscle relaxation with the contraction of tonic muscles. Relaxation periods progressively increase until the end of the ictal phase, which usually lasts no more than 1 min. The postictal phase is characterized by apathy, muscle flaccidity, and excessive salivation that can cause stridulous breathing and partial airway obstruction. Atonic seizures are characterized by sudden loss of postural muscle tone lasting 1-2 s. Consciousness is briefly affected, but there is usually no postictal confusion. Myoclonic seizures are characterized by a sudden, brief muscle contraction that may involve one part of the body or the entire body.

[006] Synaptic vesicle protein 2A ("SV2A") has been identified as a broad-spectrum anticonvulsant target in models of partial and generalized epilepsy. Studies performed in animal models and human tissue suggest that changes in SV2A expression are implicated in epilepsy (for a review see for example: (a) Mendoza-Torreblanca et al. "Synaptic vesicle protein 2A: basic facts and role in synaptic function" European Journal of Neuroscience 2013, pp. 1-11 (b) Kaminski RM, et al., "Targeting SV2A for Discovery of Antiepileptic Drugs." In: Noebels JL, Avoli M, Rogawski MA, et al., eds. Jasper's Basic Mechanisms of the Epilepsies [Internet], 4th edition. Bethesda (MD): National Center for Biotechnology Information (USA); 2012. Available from: http://www.ncbi.nlm.nih.gov/books/NBK98183/ ).

[007] The exact role of SV2A remains unclear but studies suggest that changes in SV2A expression affect synaptic function (Nowack et al. "Levetiracetam reverses synaptic deficits produced by overexpression of SV2A" PLoS One 2011, Volume 6 (12) , e29560). It has also been suggested that SV2A is a key player in exocytosis and is involved in neutrotransmission (Crowder et al. "Abnormal neurotransmission in mice lacking synaptic vesicle protein 2A (SV2A)" Proc Nat Acad Sci USA 1999, 96, pp. 15268 -15273) and studies in knockout mice suggest that the lack of SV2A results in an imbalance between glutamatergic and GABAergic neurotransmission (Venkatesan et al. "Altered balance between excitatory and inhibitory inputs onto CA pyramidal neurons from SV2A-deficient but not SV2B -deficient mice" J Neurosci Res 2012, 90, pp. 2317-2327). Decreased expression of SV2A may be a consequence of seizure activity and may be involved in the progression of epilepsy (van Vliet et al. "Decreased expression of synaptic vesicle protein 2A, the binding site for levetiracetam, during epileptogenesis and chronic epilepsy" Epilepsy 2009, 50, pp. 422-433; Feng et al. "Down-regulation of synaptic vesicle protein 2A in the anterior temporal neocortex of patients with intractable epilepsy" J Mol Neurosci 2009, 39, pp. 354-359; Toering et al "Expression patterns of synaptic vesicle protein 2A in focal cortical dysplasia and TSC-cortical tubers" Epilepsia 2009, 50, pp. 1409-1418) and epileptogenesis in patients with brain tumors (de Groot et al. "Expression of synaptic vesicle protein 2A in epilepsy-associated brain tumors and in the peritumoral cortex" Neuro-Oncology 2010, 12, pp. 265-273).

[008] SV2A binders include Levetiracetam (Lynch et al. "The synaptic vesicle protein SV2A is the binding site for the antiepileptic drug Levetiracetam" Proc. Natl. Acad. Sci. USA 2004, Vol. 101, pp. 9861-9866 ), brivaracetam, and seletracetam (Kaminski RM, et al. "Targeting SV2A for Discovery of Antiepileptic Drugs". In: Noebels JL, Avoli M, Rogawski MA, et al., eds. Jasper's Basic Mechanisms of the Epilepsies [Internet]. 4a edition. Bethesda (MD): National Center for Biotechnology Information (USA); 2012. Available from: http://www.ncbi.nlm.nih.gov/books/NBK98183/; Nowack et al. "Levetiracetam reverses synaptic deficits produced by overexpression of SV2A" PLoSone December 2011, Vol. 6 (12), e29560).

[009] Levetiracetam, (-)-(S)-α-ethyl-2-oxo-1-pyrrolidine acetamide or (S)-2-(2-oxopyrrolidin-1-yl)butanamide,

[0010] is an antiepileptic drug. It has not shown activity in traditional acute models (maximum electroshock and pentylenetetrazole seizure tests) but has been found to be potent in models of chronic epilepsy and in genetic models of generalized epilepsy. It has shown a high margin of safety compared to other antiepileptic drugs (Klitgaard "Levetiracetam: the preclinical profile of a new class of antiepileptic drugs" Epilepsia 2001, 42 (Supplement 4), pp. 13-18). It is marketed under the trade name Keppra®, available as tablets, as an oral solution, and as a concentrate made up of an infusion solution. Keppra® has been approved in Europe as a monotherapy in patients from 16 years of age with newly diagnosed epilepsy, in the treatment of partial-onset seizures (seizures) with or without secondary generalization as an adjunctive therapy for use with other drugs antiepileptics in the treatment of partial-onset seizures with or without generalization in patients from 1 month of age; myoclonic seizures in patients from 12 years of age with juvenile myoclonic epilepsy; and primary generalized tonic-clonic seizures in patients from 12 years of age with idiopathic generalized epilepsy (www.ema.europa.eu). Keppra® is approved in the US as an adjunctive therapy for the treatment of partial-onset seizures in patients 1 month of age and older; myoclonic seizures in patients 12 years of age and older with juvenile myoclonic epilepsy; and primary generalized tonic-clonic seizures in patients 6 years of age and older with idiopathic generalized epilepsy. Keppra XR®, available as extended-release tablets, is approved in the US for the adjunctive treatment of partial-onset seizures in patients 16 years of age and older with epilepsy (http://www.accessdata.fda.gov/scripts /cder/drugsatfda/index.cfm).

[0011] Brivaracetam, the 4-n-propyl analogue of levetiracetam,(2S)-2-[(4R)-oxo-4-propyl-pyrrolidin-1-yl]butanamide,

[0012] is in clinical trials and is being investigated as monotherapy in partial-onset seizures and postherpetic neuralgia and as adjunctive therapy in refractory partial-onset seizures, adolescent and adult Unverricht-Lundborg disease, and in photosensitive epilepsy (www .clinicaltrials.gov).

[0013] Seletracetam, (2S)-2-[(4S)-4-(2,2-difluorovinyl)-2-oxo-pyrrolidin-1-yl]butanamide, has been tested in clinical trials.

[0014] Processes for the preparation of the three compounds are known in the literature. For example, processes for preparing Levetiracetam are disclosed, for example, in EP 0 162 036 and GB 2 225 322. A process for preparing Brivaracetam is disclosed for example in WO 01/62726. A process for the preparation of Seletracetam is known for example from WO2005/121082. Alternative processes for preparing the three compounds are disclosed in EP1806339.

[0015] Antiepileptic drugs have found utility in neurological and psychiatric disorders, including neuropathic pain, migraine, essential tremor and in anxiety, schizophrenia and bipolar disorder (Landmarck "Antiepileptic drugs in non-epilepsy disorders. Relations between mechanisms of action and clinical efficacy " CNS Drugs 2008, Vol. 22 (1), pp. 27-47; Calabresi et al. "Antiepileptic drugs in migraine: from clinical aspects to cellular mechanisms" Trends in Pharmacological Sciences 2007, Vol. 28(4 ), pp. 188-195; Rogawski and Loscher "The neurobiology of antiepileptic drugs for the treatment of nonepileptic conditions" Nat Med 2004, Vol. 10, pp. 685-692).

[0016] Levetiracetam was found to be effective or potentially effective in a broad spectrum of neuropsychiatric disorders including mood disorders (Muralidharan and Bhagwagar "Potential of levetiracetam in mood disorders: a preliminary review" CNS Drugs 2006, Vol. 20, pp. 969 -979; Mula et al. "The role of anticonvulsant drugs in anxiety disorders: a critical review of the evidence" J Clin Pshycopharmacol 2007, Vol. 27, pp. 263-272), anxiety disorders (Kinrys et al. "Levetiracetam as adjunctive therapy for refractory anxiety disorders" J Clin Psychiatry 2007, Vol. 68, pp. 1010-1013; Zhang et al. "Levetiracetam in social phobia: a placebo controlled pilot study" J Psychopharmacol 2005, Vol. 19, pp. 551 -553; Kinrys et al. "Levetiracetam for treatment-refractory posttraumatic stress disorder" J Clin Psychiatry 2006, Vol. 67, pp. 211-214), pain (Enggaard et al. "Specific effect of levetiracetam in experimental human pain models" Eur J Pain 2006, Vol. 10, pp. 193-198; Dunteman "Levetiracetam as an adjunctive analgesic in neoplastic plexopathies: case series and commentary" J Pain Palliative Care Pharmacother 2005, Vol. 19, pp. 35-43; Price "Levetiracetam in the treatment of neuropathic pain: three case studies" Clin J Pain 2004, Vol. 20, pp. 33-36), movement disorders (Bushara et al. "The effect of levetiracetam on essential tremor" Neurology 2005, Vol. 64, pp. 1078-1080; McGavin et al "Levetiracetam as a treatment for tardive dyskinesia: a case report" Neurology 2003, Vol. 61, pp. 419; Woods et al. "Effects of levetiracetam on tardive dyskinesia: a randomized, double-blind, placebo-controlled study" J Clin Psy-chiatry 2008, Vol. 69, pp. 546-554; Zivkovic et al. "Treatment of tardive dyskinesia with levetiracetam in a transplant patient" Acta Neurol Scand 2008, Vol. 117, pp. 351-353; Striano et al. "Dramatic response to levetiracetam in post-ischaemic Holmes ' tremor" J Neurol Neurosurg Psychiatry 2007, Vol. 78, pp. 438-439) and is suspected to show potentially beneficial effects on cognitive functioning (Piazzini et al. "Levetiracetam: An improvement of attention and of oral fluency in patients with partial epilepsy" Epilepsy Research 2006, Vol. 68, pp. 181-188; de Groot et al. "Levetiracetam improves verbal memory in high-grade glioma patients" Neuro-oncology 2013, Vol. 15 (2), pp. 216223; Bakker et al. "Reduction of hippocampal hyperactivity improves cognition in amnestic mild cognitive impairment" Neuron 2012, Vol. 74, pp. 467-474; for a review: Eddy et al. "The cognitive impact of antiepileptic drugs" Ther Adv Neurol Disorder 2011, Vol. 4 (6), pp. 385,407 and references cited therein; Wheless "Levetiracetam in the treatment of childhood epilepsy" Neuropsychiatric Disease and Treatment 2007, Vol. 3(4), pp. 409-421), and behavioral symptoms in dementia (Dolder and Nealy "The efficacy and safety of newer anticonvulsants in patients with dementia" Drugs Aging 2012, Vol. 29 (8), pp. 627-637). Animal data and some preliminary clinical trials suggest that levetiracetam may have potential for restraining post-traumatic epilepsy, such as that caused by status epilepticus, traumatic brain injury and ischemic stroke, and appears to have neuroprotective effects. Levetiracetam's potential in alleviating epileptogenesis or cognitive dysfunction remains to be verified by conclusive animal and clinical studies (for reviews: Loscher and Brandt "Prevention or modification of epileptogenesis after brain insults: experimental approaches and translational research" Pharmacol Rev 2010, Vol 62, 668-700; Shetty "Prospects of levetiracetam as a neuroprotective drug against status epilepticus, traumatic brain injury and stroke" Front. Neur. 2013, 4: 172. Doi: 10.3389/fneur.2013.00172), as it exhibited activity antiepileptogenic in the kindling model in mice and rats. Levetiracetam has also been suggested to inhibit glutamate release (Lee et al. "Levetiracetam inhibits glutamate transmission through presynaptic P/Q-type calcium channels on the granule cells of the dentate gyrus" British Journal of Pharmacology 2009, Vol. 158, pp. 1753-1762).

[0017] Seletracetam and brivaracetam have been found to reduce the severity of dystonia in the dtsz mutant hamster model and may be useful in some patients suffering from dyskinetic and dystonic movement disorders (Hamann et al. "Brivaracetam and seletracetam, two new SV2A ligands, improve paroxysmal dystonia in the dtsz mutant hamster" European Journal of Pharmacology 2008, Vol. 601, pp. 99102).

[0018] Positive allosteric modulators of mGluR2 have recently emerged as promising new therapeutic approaches for the treatment of various CNS disorders, including epilepsy, and some mGluR2 AMPs are currently undergoing clinical trials for the treatment of schizophrenia, and anxious depression ( www.clinicaltrials.gov, see for example: JNJ-40411813/ADX71149 by Addex Therapeutics and Janssen Pharmaceuticals, Inc.). The initial suggestion that drugs that dampen glutamatergic transmission might be effective in the treatment of epilepsy came from acute non-clinical studies with mixed mGlu2/3 receptor agonists (Moldrich et al. "Glutamate metabotropic receptors as targets for drug therapy in epilepsy" Eur J Pharmacol 2003, Vol 476, pp. 3-16). LY379268 and LY389795, two mGlu2/3 receptor agonists, were found to be ineffective in blocking MES seizures up to doses producing motor impairment but were found to be effective in the 6 Hz model in a dose-dependent manner (Barton et al. "Comparison of the effect of glutamate receptor modulators in the 6 Hz and maximal electroshock seizure models" Epilepsy Research 2003, Vol.56, pp. 17-26). Continued administration of an mGlu2/3 agonist paradoxically induced seizure activity in long-term toxicology studies (Dunayevich et al. "Efficacy and tolerability of an mGlu2/3 agonist in the treatment of generalized anxiety disorder" Neuropsychopharmacology. 2008, Vol 33 (7), pp. 1603-10). This paradoxical effect may be related to agonist-induced changes in receptor system sensitivity (tachyphylaxis), but has not, however, been reported in preclinical models of epilepsy. Positive allosteric modulators, in contrast, modulate ongoing neurotransmission but are not directly stimulatory, thereby reducing the risk of tachyphylaxis.

[0019] Prior to seizure activity, increases in extracellular glutamate are measured in the human hippocampus and the increase is sustained during epileptogenic activity (During and Spencer "Extracellular hippocampal glutamate and spontaneous seizure in the conscious human brain" Lancet 1993, Vol. 341 (8861), pp. 1607-10), thus lending support to the idea that a reduction in glutamate levels may be beneficial in the treatment of epilepsy. In fact, during seizure activity, glutamate levels rise to potentially neurotoxic levels. Seizure activity results in progressive structural damage in the human brain further inducing abnormalities in glutamate metabolism (Petroff et al. "Glutamateglutamine cycling in the epileptic human hippocampus" Epilepsia 2002, Vol. 43(7), pp. 703-10). Thus, a positive allosteric mGluR2 modulator or an orthosteric mGluR2 agonist might be expected to protect against seizure-induced neuronal damage.

[0020] WO2009/033704 and WO2010/130424 disclose mGluR2 positive allosteric modulators, their uses and processes for synthesizing the compounds. WO1997/18199 and WO2003/104217 disclose excitatory amino acid receptor modulator compounds which were later shown to have mGlu2/3 orthosteric agonist activity (see for example Rorick-Kehn et al. (2007) The Journal of Pharmacology and Experimental therapeutics Vol 321, No. 1, pp. 308-317), additional scientific and patent literature discloses additional examples of compounds having mGlu2/3 orthosteric agonist activity, and WO2008/150233 discloses compounds with mGluR2 allosteric activating activity.

[0021] Currently available antiepileptic drugs do not merely affect glutamatergic transmission. Its mechanism of action is generally conceptualized as altering the balance between excitatory (glutamate-mediated) and inhibitory (GABA-mediated) transmission (Johannessen Landmark "Antiepileptic drugs in nonepilepsy disorders: relations between mechanisms of action and clinical efficacy" CNS Drugs 2008, Vol. 22(1), pp. 27-47).

[0022] A significant limiting factor in the use of SV2A ligands is the tolerability and side effect profile. For example, the effective dose of levetiracetam for partial-onset seizures is dosed at 1000 mg, 2000 mg, and 3000 mg given twice daily. Side effects reported for levetiracetam include aggressive or angry behavior, anxiety, change in personality, chills, coughing or hoarseness, crying, depersonalization, diarrhea, dry mouth, euphoria, fever, general feeling of discomfort or illness, headache, hyperventilation, irregular heartbeat, irritability, joint pain, loss of appetite, lower back or side pain, mental depression, muscle aches and pains, nausea, painful or difficult urination, paranoia, quick to react or overreact emotionally, mood swings that change quickly, restlessness, agitation, tremors, shortness of breath, unusual sleepiness or drowsiness, sore throat, stuffy or runny nose, sweating, difficulty sleeping, unusual tiredness or weakness, and vomiting. Thus there is still a need to provide effective treatment with a lower effective dose of levetiracetam and a more favorable side effect profile for the treatment of epilepsy and related disorders, not only in the adult population, but also in the pediatric population.

BRIEF DESCRIPTION OF THE FIGURES

[0023] Figure 1: Dose response for the determination of ED50 of 44mA 6 Hz for Co. At the. 2 and LEV alone and in combination.

[0024] Figure 2: Isobolographic analysis for the combination of Co. At the. 1 with levetiracetam (LEV) in the 6 Hz assay (44 mA). Initial ED50 values (shown below) were determined for both Co. At the. 1 as for LEV (data points on x and y axes; full diamonds). The theoretical additivity line connects the calculated ED50 values for the two compounds (solid black line). The theoretical ED50 (+ SEM) for three fixed ratio dose combinations (LEV: Co. No. 1) are plotted: 1:3 - filled squares/solid black line, 1:1 - filled right triangles/solid black line, and 3:1 - full inverted triangles/solid black line. Experimental treatment doses were initially derived from theoretical values and adjusted according to observed effects. The experimentally determined ED50 values (+ SEM) for each dose-fixed ratio combination are also shown: 1:3' - open squares/dotted line, 1:1' - right triangles/dotted line, and 3:1' - open inverted triangles/dotted line. Comparisons between theoretical and experimentally determined ED50 values were compared using a t-test (***P<0.001). N = 8 per group. In Figure 2, the ratio of LEV to Co. At the. 1 is illustrated as follows:

[0025] Figure 3: Combination Studies for Co. At the. 25-a with Levetiracetam (LEV) in the 6 Hz Assay (44 mA). At a dose of 10 mg/kg s.c., Co. At the. 25-a increases the power of LEV, leading to an approximately 70-fold deviation in ED50. This suggests a positive pharmacodynamic relationship.

[0026] Figure 4: Combination Studies for Co. At the. 2-a with Levetiracetam (LEV) in the 6 Hz Assay (44 mA). At a dose of 10 mg/kg s.c., Co. At the. 2-a increases LEV power, leading to an approximately 35-fold shift in ED50. This suggests a positive pharmacodynamic relationship.

[0027] Figure 5: Combination Studies for Co. At the. 6-b with Levetiracetam (LEV) in the 6 Hz Assay (44 mA). At a dose of 10 mg/kg p.o., Co. At the. 6-b increases LEV power, leading to an approximately 100-fold shift in ED50. This suggests a positive pharmacodynamic relationship.

[0028] Figure 6: Combination Studies for LY-404039 with Levetiracetam (LEV) in the 6 Hz Assay (44 mA). At a dose of 5 mg/kg s.c., LY-404039 increases LEV potency, leading to an approximately 27-fold shift in ED50. This suggests a positive pharmacodynamic relationship.

DESCRIPTION OF THE INVENTION

[0029] The present invention relates to a combination comprising (a) a synaptic vesicle protein 2A ("SV2A") ligand; and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof.

[0030] In a particular embodiment, the invention as described herein relates to a pharmaceutical combination, in particular a pharmaceutical combination product, comprising (a) a synaptic vesicle protein 2A ("SV2A") ligand; and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof; and (c) at least one pharmaceutically acceptable carrier.

[0031] In a further embodiment, the invention relates to the combination described herein for use as a medicine.

[0032] A further embodiment of this invention relates to the use of the combination described herein for the manufacture of a medicament or a pharmaceutical product for the treatment or prevention of epilepsy and related disorders; neuropathic pain; migraine or resistant headache and bipolar and related disorders.

[0033] A further embodiment of this invention relates to the use of the combination described herein for the manufacture of a medicament or a pharmaceutical for neuroprotection.

[0034] A further embodiment of this invention relates to the use of the combination described herein for the manufacture of a medicament or a pharmaceutical product for the prevention of epileptogenesis.

[0035] An additional modality relates to the treatment or prevention of epilepsy and related disorders; neuropathic pain; migraine or resistant headache; and bipolar and related disorders of a subject comprising administering simultaneously or sequentially to the subject in need of a synaptic vesicle protein 2A ("SV2A" ligand); and a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound or a salt thereof pharmaceutically acceptable compound or solvate, in amounts that would be therapeutically effective when the SV2A ligand and the mGluR2 compound are administered together.

[0036] An additional embodiment relates to a combination as described here for neuroprotection; or with a combination as described herein for use in neuroprotection.

[0037] An additional modality relates to a combination as described here for the prevention of epileptogenesis; or with a combination as described herein for use in preventing epileptogenesis.

[0038] In a further embodiment, the invention relates to a method of treating or preventing epilepsy and related disorders; neuropathic pain; migraine or resistant headache; bipolar and related disorders in patients comprising administering a fixed dose combination of (a) a synaptic vesicle protein 2A ("SV2A" ligand); and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof,

[0039] in amounts that would be therapeutically effective when the SV2A ligand and the mGluR2 compound are administered together.

[0040] In a further embodiment, the invention relates to a method of neuroprotection with a combination as defined herein.

[0041] In a further embodiment, the invention relates to a method of anti-epileptogenesis with a combination as defined herein.

[0042] An additional embodiment relates to a method for treating or preventing epilepsy and related disorders; neuropathic pain; migraine or resistant headache; bipolar and related disorders, said method comprising administering a therapeutically effective amount of a combination or a combination product comprising (a) a synaptic vesicle protein 2A ("SV2A" ligand); and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof,

[0043] to an individual with its need, such as a warm-blooded animal, in particular a human.

[0044] An additional embodiment relates to a method of neuroprotection, said method comprising administering a therapeutically effective amount of a combination or a combination product comprising (a) a synaptic vesicle protein 2A ("SV2A" ligand); and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof,

[0045] to an individual with its need, such as a warm-blooded animal, in particular a human.

[0046] A further embodiment relates to a method of antiepileptogenesis, said method comprising administering a therapeutically effective amount of a combination or a combination product comprising (a) a synaptic vesicle protein 2A ("SV2A" ligand); and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof,

[0047] to an individual with its need, such as a warm-blooded animal, in particular a human.

[0048] In a further embodiment, the present invention relates to a pharmaceutical product or a commercial package comprising a combination according to the invention as described herein, in particular together with instructions, for its simultaneous, separate or sequential use in treatment or prevention of epilepsy and related disorders; neuropathic pain; migraine or resistant headache; and related disorders.

[0049] In a further embodiment, the present invention relates to a pharmaceutical product or a commercial package comprising a combination according to the invention as described herein, in particular together with instructions, for their simultaneous, separate or sequential use in neuroprotection.

[0050] In a further embodiment, the present invention relates to a pharmaceutical product or a commercial package comprising a combination according to the invention as described herein, in particular together with instructions, for their simultaneous, separate or sequential use in anti-epileptogenesis .

[0051] In a further embodiment, the invention relates to a combination comprising an amount that is jointly therapeutically effective against epilepsy and related disorders; neuropathic pain; migraine or resistant headache; bipolar and related disorders; of (a) a synaptic vesicle protein 2A ("SV2A") ligand; and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable carrier.

[0052] In a further embodiment, the invention relates to a combination comprising an amount that is jointly therapeutically effective and neuroprotective, of (a) a ligand for synaptic vesicle protein 2A ("SV2A"); and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable carrier.

[0053] In a further embodiment, the invention relates to a combination comprising an amount that is jointly therapeutically effective in preventing epileptogenesis, of (a) a synaptic vesicle protein 2A ("SV2A") ligand; and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof, and at least one pharmaceutically acceptable carrier.

[0054] In a further embodiment, the invention relates to the use of (a) a synaptic vesicle protein 2A ("SV2A") ligand; and (b) a metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound, or a pharmaceutically acceptable salt or solvate thereof, or a metabotropic glutamatergic receptor subtype 2 orthosteric agonist compound, or a pharmaceutically acceptable salt or solvate thereof,

[0055] for preparing a combination product according to the present invention.

[0056] The components (b) of the combination of the invention are generally referred to herein as "mGluR2 compounds" or "PAM compounds/mGluR2 agonists", or "mGluR2 positive allosteric modulator/mGluR2 orthosteric agonist compounds" meaning that the compounds have primarily activity at the metabotropic glutamatergic receptor subtype 2, and are in particular selected from positive allosteric modulators (PAMs) of the metabotropic glutamatergic receptor subtype 2, and orthosteric agonists of the metabotropic glutamatergic receptor subtype 2. the skilled person will be familiar with the great homology of mGluR2 and mGluR3, due to which some orthosteric mGluR2 agonists also exhibit activity as orthosteric mGluR3 agonists. Such is the case, for example, of (-)-(1R,4S,5S,6S)-4-amino-2-sulfonylbicyclo[3.1.0]hexane-4,6-dicarboxylic acid (also known as LY- 404,039 [CAS 635318-115]), with a Ki = 149 nM (mGlu2 receptor) and Ki = 92 nM (mGlu3 receptor), 100-fold selectivity for mGlu2 and mGlu3 over mGlu4a, -6, -7a , and -8a, and no activity on mGlu1a and mGlu5a ( Rorick-Kehn et al. (2007) The Journal of Pharmacology and Experimental Therapeutics Vol. 321, No. 1, pp. 308-317). The term "mGluR2 compounds" or "PAM/mGluR2 agonist compounds", or "mGluR2 positive allosteric modulator/mGluR2 orthosteric agonist compound" does not, therefore, exclude compounds exhibiting some other additional minimal in vitro or in vivo activity. .

[0057] The mGluR2 PAM compounds of the combination of the invention are in particular selected from those disclosed in WO2010/130424. A particular subgroup of said compounds disclosed in WO2010/130424 can be defined by the following Formula (I)

[0058] or a stereoisomeric form thereof; on what

[0059] R1 is selected from the group consisting of (C3-7 cycloalkyl)C1-3 alkyl-, mono- or polyhaloC1-4 alkyl, and (C1-4 alkyl)-O-(C1-4 alkyl);

[0060] R2 is halo or C1-4 polyhaloalkyl;

[0061] A is a covalent bond or a -CH2-;

[0062] L is selected from radicals (a), (b) and (c):

[0063] in which

[0064] R3a is selected from unsubstituted phenyl or phenyl substituted by 1 or 2 halo substituents;

[0065] R4a is selected from the group of hydrogen, C1-3 alkyl, and ha lo;

[0066] or R3a-C-R4a together represent a radical of the formula (a-1)

[0067] wherein R5a is hydrogen or halo;

[0068] R3b is selected from the group of phenyl substituted by 1 or 2 halo substituents, pyridinyl substituted by 1 or 2 halo substituents, unsubstituted pyrimidinyl and pyrimidinyl substituted by 1 or 2 C1-3alkyloxy substituents;

[0069] or a pharmaceutically acceptable salt or solvate thereof.

[0070] Thus, according to a particular embodiment of the invention, the metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound is a compound of Formula (I) as defined herein.

[0071] In a particular embodiment, the compounds of Formula (I) are as defined herein wherein

[0072] R1 is selected from the group consisting of cyclopropylmethyl-, 2,2,2-trifluoroethyl, and CH3-O-CH2-;

[0073] R2 is chlorine or CF3;

[0074] A is a covalent bond or a -CH2-;

[0075] L is selected from radicals (a), (b) and (c):

[0076] in which

[0077] R3a is selected from unsubstituted phenyl or phenyl substituted by 1 or 2 fluorine substituents;

[0078] R4a is selected from the group of hydrogen, methyl and fluorine;

[0079] or R3a-C-R4a together represent a radical of the formula (a-1)

[0080] wherein R5a is hydrogen or fluorine;

[0081] R3b is selected from the group of phenyl substituted by 1 or 2 fluorine substituents, pyridinyl substituted by 1 or 2 fluorine substituents, unsubstituted pyrimidinyl and pyrimidinyl substituted by 1 or 2 methoxy substituents;

[0082] or a pharmaceutically acceptable salt or solvate thereof.

[0083] In a particular embodiment, the compounds of Formula (1) are as defined herein wherein (i) when A is CH2; and R2 is trifluoromethyl; then

[0084] R1 is cyclopropylmethyl-; It is

[0085] L is selected from (ii) when A is CH2; and R2 is chloro; then

[0086] R1 is cyclopropylmethyl-; It is

[0087] L is (iii) when A is a covalent bond; and R2 is trifluoromethyl; then

[0088] R1 is cyclopropylmethyl; It is

[0089] L is selected from (iv) when A is a covalent bond and R2 is Cl; then (iv-a) R1 is cyclopropylmethyl and L is

[0090] or (iv-b) R1 is 2,2,2-trifluoroethyl and L is selected from (v) when A is CH2 and R1 is -CH2-O-CH3; then

[0091] R2 is -CF3 and L is

[0092] or a pharmaceutically acceptable salt or solvate thereof.

[0093] The compounds of Formula (I) are disclosed in WO2010/130424 and can be prepared according to the processes described herein, which are hereby incorporated by reference in their entirety.

[0094] Particular compounds of Formula (I) include

[0095] In one embodiment of the invention, the compound of Formula (I) is Co. At the. 1; or a pharmaceutically acceptable salt thereof, preferably a hydrochloride salt thereof.

[0096] In an additional embodiment of the invention, the compound of Formula (I) is Co. At the. two; or a pharmaceutically acceptable salt thereof, preferably a hydrochloride (.HCl) salt thereof.

[0097] The mGluR2 PAM compounds of the combination of the invention are also in particular selected from those disclosed in WO2009/033704. Said compounds disclosed in WO2009/033704 can be defined by the following Formula (IA)

[0098] and their stereochemically isomeric forms, in which

[0099] R1 is C1-6 alkyl; or C 1-3 alkyl substituted by C 3-7 cycloalkyl, phenyl, or phenyl substituted by halo, trifluoromethyl or trifluoromethoxy;

[00100] R2 is halo, trifluoromethyl, C1-3 alkyl or cyclopropyl;

[00101] R3 is hydrogen, fluorine, hydroxyl, C1-3 hydroxyalkyl, C1-3 hydroxyalkyloxy, C1-3 fluoroalkyl, C1-3fluoroalkyloxy or cyano; It is

[00102] Ar is unsubstituted phenyl; or phenyl substituted by n radicals R4, where n is 1, 2 or 3;

[00103] R4 is selected from the group consisting of hydrogen, halo, C1-3 alkyl, C1-3 hydroxyalkyl, C1-3 polyhaloalkyl, cyano, hydroxyl, amino, carboxyl, C1-3 alkyloxyC1-3 alkyl, C1-3 alkyloxy, polyhaloC1-3alkyloxy, C1-3alkylcarbonyl, mono- and di(C1-3alkyl)amino, and morpholinyl; or

[00104] two vicinal R4 radicals taken together form a bivalent radical of the formula -N=CH-NH- (i), -CH=CH-NH- (ii), or -O-CH2-CH2-NH- (iii ); or

[00105] R3 and a radical R4 in ortho position taken together form a bivalent radical of the formula -CH2-O- (iv), or -O-CH2- (v);

[00106] and their pharmaceutically acceptable salts and solvates.

[00107] In a particular embodiment, the compounds of Formula (I-A) are as defined herein wherein

[00108] R1 is C1-6 alkyl; or C 1-3 alkyl substituted by C 3-7 cycloalkyl, phenyl, or phenyl substituted by halo, trifluoromethyl or trifluoromethoxy;

[00109] R2 is halo, trifluoromethyl, C1-3 alkyl or cyclopropyl;

[00110] R3 is hydrogen, fluorine, hydroxyl, C1-3 hydroxyalkyl, C1-3 hydroxyalkyloxy, C1-3 fluoroalkyl, C1-3 fluoroalkyloxy or cyano; It is

[00111] Ar is unsubstituted phenyl, or phenyl substituted by n radicals R4, wherein n is 1, 2 or 3;

[00112] R4 is selected from the group consisting of hydrogen, halo, C1-3 alkyl, C1-3 hydroxyalkyl, C1-3 polyhaloalkyl, cyano, hydroxyl, amino, carboxyl, C1-3 alkyloxyC1-3 alkyl, C1-3 alkyloxy, C1-3oxy polyhaloalkyl; C1-3alkylcarbonyl, mono- and di(C1-3alkyl)amino, and morpholinyl; or

[00113] two vicinal R4 radicals taken together form a bivalent radical of the formula -N=CH-NH- (i), -CH=CH-NH- (ii), or -O-CH2-CH2-NH- (iii );

[00114] and their pharmaceutically acceptable salts and solvates.

[00115] In a particular embodiment, the compounds of Formula (I-A) are as defined herein wherein

[00116] R1 is C1-6 alkyl; or C 1-3 alkyl substituted by C 3-7 cycloalkyl, phenyl or phenyl substituted by halo, trifluoromethyl or trifluoromethoxy;

[00117] R2 is halo, trifluoromethyl, C1-3 alkyl or cyclopropyl;

[00118] R3 is hydrogen, fluorine, hydroxyl, C1-3 hydroxyalkyl, C1-3 hydroxyalkyloxy, C1-3 fluoroalkyl, C1-3 fluoroalkyloxy or cyano; It is

[00119] Ar is unsubstituted phenyl;

[00120] and their pharmaceutically acceptable salts and solvates.

[00121] In a further embodiment, compounds of Formula (IA) are as defined herein wherein

[00122] R1 is 1-butyl, 2-methyl-1-propyl, 3-methyl-1-butyl, (cyclopropyl)methyl or 2-(cyclopropyl)-3-ethyl;

[00123] R3 is hydrogen, fluorine or cyano; It is

[00124] Ar is unsubstituted phenyl;

[00125] and their pharmaceutically acceptable salts and solvates.

[00126] In a further embodiment, compounds of Formula (IA) are as defined herein wherein

[00127] R1 is 1-butyl, 3-methyl-1-butyl, (cyclopropyl)methyl or 2-(cyclopropyl)-1-ethyl;

[00128] R2 is chlorine;

[00129] R3 is hydrogen or fluorine; It is

[00130] Ar is unsubstituted phenyl;

[00131] and their pharmaceutically acceptable salts and solvates.

[00132] In a further embodiment, compounds of Formula (IA) are as defined herein wherein

[00133] R1 is C1-6 alkyl; or C 1-3 alkyl substituted by C 3-7 cycloalkyl, phenyl, or phenyl substituted by halo, trifluoromethyl or trifluoromethoxy;

[00134] R2 is halo, trifluoromethyl, C1-3 alkyl or cyclopropyl;

[00135] R3 is hydrogen, fluorine, hydroxyl, C1-3 hydroxyalkyl, hydroxy C1-3 alkyloxy, C1-3 fluoroalkyl, C1-3 fluoroalkyl oxy or cyano; It is

[00136] Ar is phenyl substituted by n radicals R4, wherein n is 1, 2, or 3;

[00137] R4 is selected from the group consisting of halo, C1-3 alkyl, C1-3 hydroxyalkyl, C1-3 alkyloxy, polyhaloC1-3 alkyloxy, C1-3 alkylcarbonyl, mono- and di(C1-3 alkyl)amino , and morpholinyl; or

[00138] two vicinal R4 radicals taken together form a bivalent radical of the formula -N=CH-NH- (i), -CH=CH-NH- (ii), or -O-CH2-CH2-NH- (iii ); or

[00139] R3 and a radical R4 in ortho position taken together form a bivalent radical of the formula -CH2-O- (iv), or -O-CH2- (v);

[00140] and their pharmaceutically acceptable salts and solvates.

[00141] In a further embodiment, compounds of Formula (IA) are as defined herein wherein

[00142] R1 is 1-butyl, 2-methyl-1-propyl, 3-methyl-1-butyl, (cyclopropyl)methyl or 2-(cyclopropyl)-1-ethyl;

[00143] R3 is hydrogen, fluorine or cyano; It is

[00144] Ar is phenyl substituted by halo, trifluoromethyl, morpholinyl or hydroxyC1-3alkyl;

[00145] and their pharmaceutically acceptable salts and solvates.

[00146] In a further embodiment, compounds of Formula (IA) are as defined herein wherein

[00147] R1 is 1-butyl, 3-methyl-1-butyl, (cyclopropyl)methyl or 2-(cyclopropyl)-1-ethyl;

[00148] R2 is chlorine;

[00149] R3 is hydrogen or fluorine; It is

[00150] Ar is phenyl substituted by at least one halo group;

[00151] and their pharmaceutically acceptable salts and solvates.

[00152] In a further embodiment, the compounds of Formula (I-A) are as defined herein wherein

[00153] R1 is 1-butyl, 3-methyl-1-butyl, (cyclopropyl)methyl or 2-(cyclopropyl)-1-ethyl;

[00154] R2 is chlorine;

[00155] R3 is hydrogen or fluorine; It is

[00156] Ar is phenyl substituted by at least two fluorine groups;

[00157] and their pharmaceutically acceptable salts and solvates.

[00158] The compounds of Formula (I-A) are disclosed in WO2009/033704 and can be prepared according to the processes described herein, which are hereby incorporated by reference in their entirety.

[00159] Particular compounds of Formula (IA) include

[00160] and their pharmaceutically acceptable salts and solvates.

[00161] In one embodiment of the invention, the compound of Formula (IA) is

[00162] or a pharmaceutically acceptable salt or solvate thereof.

[00163] The mGluR2 PAM compounds of the combination of the invention are also in particular selected from those disclosed in PCT/EP2014/068676. Said compounds disclosed in PCT/EP2014/068676 can be defined by the following Formula (IB)

[00164] and their stereochemically isomeric forms, in which

[00165] R1 is selected from the group consisting of C1-6 alkyl, (C3-8 cycloalkyl)C1-3 alkyl, and (C1-3 alkyloxy)C1-3 alkyl;

[00166] each R2 is independently selected from F, Cl, C1-3 alkyl, C1-3 alkyloxy, mono- or polyhaloC1-3 alkyl, and mono- or polyhalo-C1-3 alkyloxy;

[00167] n is an integer selected from 1, 2, and 3;

[00168] and their pharmaceutically acceptable salts and solvates.

[00169] The mGluR2 PAM compounds of the combination of the invention are in particular selected from compounds of Formula (I-B) as defined above, and their stereoisomeric forms, wherein R1 is selected from the group consisting of CH3CH2, CH3CH2CH2, (cyclopropyl)methyl, (cyclobutyl)methyl, ethyloxymethyl and methyloxymethyl; and the rest of the variables are as defined here; and their pharmaceutically acceptable salts and solvates.

[00170] In a further embodiment, the mGluR2 PAM compounds of the combination of the invention are in particular selected from compounds of Formula (I-B) as defined above, and their stereoisomeric forms, wherein R1 is selected from the group consisting of CH3CH2, (cyclopropyl )methyl, (cyclobutyl)methyl and methyloxymethyl; and the rest of the variables are as defined here; and their pharmaceutically acceptable salts and solvates.

[00171] In a further embodiment, the mGluR2 PAM compounds of the combination of the invention are in particular selected from compounds of Formula (I-B) as defined above, and their stereoisomeric forms, wherein R1 is selected from the group consisting of CH3CH2, (cyclopropyl )methyl, (cyclobutyl)methyl and ethyloxymethyl; and the rest of the variables are as defined here; and their pharmaceutically acceptable salts and solvates.

[00172] Thus, according to a particular embodiment of the invention, the metabotropic glutamatergic receptor subtype 2 ("mGluR2") positive allosteric modulator ("PAM") compound is a compound of Formula (I-B) as defined herein.

[00173] In a further embodiment, the compounds of Formula (IB) are as defined herein wherein

[00174] each R2 is independently selected from F, Cl, CH3, CH3O and CF3; and their pharmaceutically acceptable salts and solvates.

[00175] In a further embodiment, compounds of Formula (IB) are as defined herein having Formula (I-Ba)

[00176] wherein the variables are as defined in Formula (I-B) herein, and pharmaceutically acceptable salts and solvates thereof.

[00177] In a further embodiment, compounds of Formula (IB) are as defined herein having Formula (I-Bb)

[00178] wherein the variables herein, and their pharmaceutically acceptable salts and solvates.

[00179] Particular compounds of formula (I-B) include 3-(Cyclopropylmethyl)-7-[1-(4-fluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a] ]pyridine; 3-(Cyclopropylmethyl)-7-[(1*R)-1-(4-fluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1*S)-1-(4-fluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(2,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1R)-1-(2,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[1-(2,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(3,5-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(3,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(2,3-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(2,5-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(2,6-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(4-fluoro-2-methoxyphenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclobutylmethyl)-7-[1-(2,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 7-[(1S)-1-(2-Chloro-4-methylphenoxy)ethyl]-3-(cyclopropylmethyl)-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-7-[(1S)-1-(4-fluoro-2-methylphenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclopropylmethyl)-8-(trifluoromethyl)-7-[(1S)-1-(2,4,6-trifluorophenoxy)ethyl][1,2,4]triazolo[4,3-a]pyridine; 7-[1-(2,4-Difluorophenoxy)ethyl]-3-(ethoxymethyl)-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-Ethyl-8-(trifluoromethyl)-7-[1-(2,4,6-trifluorophenoxy)ethyl][1,2,4]triazolo[4,3-a]pyridine; 7-[1-(2,4-Difluorophenoxy)ethyl]-3-ethyl-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclobutylmethyl)-7-[(1*R)-1-(2,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Cyclobutylmethyl)-7-[(1*S)-1-(2,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-(Ethoxymethyl)-8-(trifluoromethyl)-7-[(1*R)-1-(2,4,6-trifluorophenoxy)ethyl][1,2,4]triazolo[4,3-a]pyridine ; 3-(Ethoxymethyl)-8-(trifluoromethyl)-7-[(1*S)-1-(2,4,6-trifluorophenoxy)ethyl][1,2,4]triazolo[4,3-a]pyridine ; 7-[(1*S)-1-(2,4-Difluorophenoxy)ethyl]-3-(ethoxymethyl)-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 7-[(1*R)-1-(2,4-Difluorophenoxy)ethyl]-3-(ethoxymethyl)-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 7-[(1*R)-1-(2,4-Difluorophenoxy)ethyl]-3-ethyl-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 7-[(1*S)-1-(2,4-Difluorophenoxy)ethyl]-3-ethyl-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 7-[1-(2,4-Difluorophenoxy)ethyl]-3-propyl-8-(trifluoromethyl)[1,2,4]triazolo[4,3-a]pyridine; 3-Ethyl-8-(trifluoromethyl)-7-[(1*R)-1-(2,4,6-trifluorophenoxy)ethyl]-[1,2,4]triazolo[4,3-a]pyridine ; 3-Ethyl-8-(trifluoromethyl)-7-[(1*S)-1-(2,4,6-trifluorophenoxy)ethyl]-[1,2,4]triazolo[4,3-a]pyridine ; 7-[(1*R)-(2,4-difluorophenoxy)ethyl]-3-propyl-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridine; and 7-[(1*S)-(2,4-difluorophenoxy)ethyl]-3-propyl-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridine.

[00180] Included within the scope of this list are their stereoisomeric forms, pharmaceutically acceptable salts and solvates.

[00181] In an additional embodiment, the compound can be selected from 3-(Cyclopropylmethyl)-7-[(1S)-1-(2,4-difluorophenoxy)ethyl]-8-(trifluoromethyl)[1] hydrochloride salt ,2,4]triazolo[4,3-a]pyridine.

[00182] The mGluR2/mGluR2/3 orthosteric agonists of the combination of the invention include, but are not limited to, for example, LY-404039; LY-2969822; LY-2934747; LY-379268; DCG-IV; LY-354740; LY-314582; LY-544344; LY-2140023;

[00183] LY-181837; LY-389795; LY-446433; LY-450477; LY-395756; LY-566332; LY-541850; LY-2300559; LY-404040; LY-281223; LY-2979165; talaglumetad; MGS008; MGS0022; MGS0028; MGS0039;

[00184] (-)-2-oxa-4-aminobicyclo[3.1.0]hexane-4,6-dicarboxylate; (+)-4-amino-2-sulfonylbicyclo[3.1.0]hexane-4,6-dicarboxylic acid; (+)-2-amino-4-fluorobicyclo-[3.1.0]hexane-2,6-dicarboxylic acid; 1S,2R,5S,6S-2-amino-6-fluoro-4-oxobicyclo-[3.1.0]hexane-2,6-dicarboxylic acid; 1S,2R,4S,5S,6S-2-amino-6-fluoro-4-hydroxybicyclo[3.1.0]hexane-2,6-dicarboxylic acid; 1S,2R,3R,5S,6S-2-amino-3-fluorobicyclo[3.1.0]hexane-2,6-dicarboxylic acid; 1S,2R,3S,5S,6S-2-amino-6-fluoro-3-hydroxybicyclo[3.1.0]hexane-2,6-dicarboxylic acid; (+)-4-amino-2-sulfonylbicyclo-[3.1.0]hexane-4,6-dicarboxylic acid; (+)-2-amino-4-fluorobicyclo[3.1.0]hexane-2,6-dicarboxylic acid; 1S,2R,5S,6S-2-amino-6-fluoro-4-oxobicyclo[3.1.0]hexane-2,6-dicarboxylic acid; 1S,2R,4S,5S,6S-2-amino-6-fluoro-4-hydroxybicyclo[3.1.0]hexane-2,6-dicarboxylic acid; 1S,2R,3R,5S,6S-2-amino-3-fluorobicyclo[3.1.0]hexane-2,6-dicarboxylic acid; or 1S,2R,3S,5S,6S-2-amino-6-fluoro-3-hydroxybicyclo-[3.1.0]hexane-2,6-dicarboxylic acid.

[00185] A particular group of mGluR2 agonists include LY-379268; DCG-IV; LY-354740; LY-404039; LY-2969822; LY-2934747; LY-544344; and LY-2140023.

[00186] The metabotropic glutamatergic receptor subtype 2 orthosteric agonists of the combination of the invention are in particular additionally selected from those disclosed in WO1997/18199 and WO2003/104217, incorporated herein in their entirety. Particular compounds disclosed therein are (-)-(1R,4S,5S,6S)-4-amino-2-sulfonylbicyclo[3.1.0]hexane-4,6-dicarboxylic acid (also known as LY-404039)

[00187] or a salt or solvate thereof, and (1R,4S,5S,6S)-4-[[(2S)-2-amino-4-(methylthio)-1-oxobutyl] acid 2,2-dioxide amino]-2-thiabicyclo[3.1.0]hexane-4,6-dicarboxylic acid (also known as LY-2140023 [CAS 635318-55-7])

[00188] or a salt or solvate thereof, for example its monohydrate.

[00189] The names of the compounds of the present invention were generated according to the nomenclature rules agreed upon by the Chemical Abstracts Service (C.A.S.) using software from Advanced Chemical Development, Inc. (ACD/Product Version Name 10.01.0.14105, October 2006). In the case of tautomeric forms, the name of the tautomeric form of the illustrated structure was generated. However, it should be clear that the other tautomeric form not illustrated is also included within the scope of the present invention.

[00190] As used herein, the notation "C1-3 alkyl", "C1-4 alkyl" or "C1-6 alkyl" as a group or part of a group defines a hydrocarbon radical, straight or branched, saturated having to 1 to 3 or 1 to 4 or 1 to 6 carbon atoms, such as methyl, ethyl, 1-propyl, 1-methylethyl, butyl, 1-methylpropyl, 2-methyl-1-propyl, 1,1-dimethylethyl , 3-methyl-1-butyl, 1-pentyl, 1-hexyl and the like.

[00191] The notation "C3-7 cycloalkyl" or "C3-8 cycloalkyl" as a group or part of a group defines a cyclic, saturated hydrocarbon radical having from 3 to 7 or from 3 to 8 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[00192] The notation "halo" or "halogen" as used herein as a group or part of a group refers to fluorine, chlorine, bromine or iodine, with fluorine or chlorine being preferred.

[00193] The notation "mono- and polyhaloalkyl C1-3" or "mono- and polyhaloalkyl C1-4" shall denote C1-3 alkyl or C1-4 alkyl, respectively, as defined above, replaced by 1, 2, 3 or where possible by more halo atoms as defined above.

[00194] Whenever the term "substituted" is used in the present invention, it is intended, unless otherwise indicated or it is clear from the context, to indicate that one or more hydrogens, preferably from 1 to 3 hydrogens, more preferably from 1 to 2 hydrogens, more preferably 1 hydrogen, on the atom or radical indicated in the expression using "substituted" are replaced by a selection from the group indicated, provided that the normal valence is not exceeded, and that the substitution results in a chemically stable, i.e., a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into a therapeutic agent.

[00195] As used herein, unless otherwise noted, the term "antiepileptic agent" and the abbreviation "AED" shall be used interchangeably with the term "anticonvulsant agent", and as used herein, refers to an agent capable of to treat, inhibit, or prevent seizure activity or ichtogenesis when the agent is administered to a subject or patient.

[00196] As used herein, unless otherwise noted, the term "synaptic vesicle protein 2A ligand" and the abbreviation "SV2A ligand" shall be used interchangeably. Examples of SV2A ligands include, but are not limited to, compounds included in GB publications 1,039,113; GB 1,309,692; EP 1 262 036; EP 1 806 339; WO 2001/062726; US 2002/094787; WO 2004/087658; WO 2005/121082; WO 2005/054188; WO 2006/128692; WO 2006/128693; WO 2007/065595; WO 2008/132139, and WO 2008/132142; WO 2011/047860; WO 2012/143116; and WO 2012/143117. Suitable particular examples of SV2A binders include, but are not limited to: levetiracetam, brivaracetam and seletracetam.

[00197] Therefore, in one embodiment of the invention, the SV2A ligand is selected from levetiracetam, brivaracetam and seletracetam.

[00198] In a particular embodiment, the SV2A ligand is leveti-racetam.

[00199] In a particular embodiment, the SV2A ligand is brivaractam.

[00200] Processes for the preparation of the above SV2A ligands are known from the literature and described for example in EP 1 806 339; EP 0 162 036 and GB 2 225 322 (levetiracetam); in WO 01/62726 (brivaracetam); and WO 2005/121082 (seletracetam); such processes are hereby incorporated by reference in their entirety.

[00201] In a further embodiment, the combination according to the invention comprises (a) an SV2A binder selected from levetiracetam or brivaracetam; and (b)

[00202] or a pharmaceutically acceptable salt thereof, preferably a hydrochloride salt thereof, or a solvate thereof.

[00203] In a further embodiment, the pharmaceutical composition according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of

[00204] or a pharmaceutically acceptable salt thereof, preferably a hydrochloride salt thereof, or a solvate thereof

[00205] In a further embodiment, the combination according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of

[00206] or a pharmaceutically acceptable salt, or solvate thereof.

[00207] In a further embodiment, the pharmaceutical composition according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of

[00208] or a pharmaceutically acceptable salt, or solvate thereof.

[00209] In a further embodiment, the combination according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of

[00210] or a pharmaceutically acceptable salt, or solvate thereof.

[00211] In a further embodiment, the pharmaceutical composition according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of

[00212] or a pharmaceutically acceptable salt, or solvate thereof.

[00213] In a further embodiment, the combination according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of

[00214] or a pharmaceutically acceptable salt thereof, in particular its hydrochloride salt, or a solvate thereof.

[00215] In a further embodiment, the pharmaceutical composition according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of

[00216] or a pharmaceutically acceptable salt thereof, in particular its hydrochloride salt, or a solvate thereof.

[00217] In a further embodiment, the combination according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of LY-404039 or a pharmaceutically acceptable salt thereof, in particular its hydrochloride salt, or a solvate thereof.

[00218] In a further embodiment, the pharmaceutical composition according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of LY-404039 or a pharmaceutically acceptable salt thereof, in particular its hydrochloride salt, or a solvate thereof.

[00219] In a further embodiment, the combination according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of LY-2140023 or a pharmaceutically acceptable salt or solvate thereof, in particular its monohydrate.

[00220] In a further embodiment, the pharmaceutical composition according to the invention comprises (a) a therapeutically effective amount of levetiracetam or brivaracetam; and (b) a pharmaceutically effective amount of LY-2140023 or a pharmaceutically acceptable salt or solvate thereof, in particular its monohydrate.

[00221] The combination product of the present invention, in particular the pharmaceutical composition according to the invention, is especially suitable for the treatment of epilepsy and related disorders.

It will be appreciated that some of the mGluR2 compounds, in particular the mGluR2 PAM compounds/agonists of the invention and their pharmaceutically acceptable addition salts and solvates, may contain one or more centers of chirality and exist as stereoisomeric forms.

[00223] The term "compounds of the invention", as used herein, is intended to include mGluR2 PAM compounds, in particular compounds of Formula (I)/(I-A)/(I-B), and mGluR2 agonist compounds such as disclosed herein, and their salts and solvates.

[00224] As used herein, any chemical formula with bonds shown only as solid lines and not as solid punched or punched punched bonds, or otherwise indicated as having a particular configuration (eg, R, S) around one or more atoms, contemplates every possible stereoisomer, or mixture of two or more stereoisomers.

[00225] Hereinafter and hereinafter, the terms "mGluR2 compound" and "PAM compound/mGluR2 agonist" are intended to include both their stereoisomers and their tautomeric forms. The terms "stereoisomers", "stereoisomeric forms" or "stereochemically isomeric forms" hereinafter or hereinafter are used interchangeably. The invention includes all stereoisomers of the compounds of the invention either as a pure stereoisomer or as a mixture of two or more stereoisomers. Enantiomers are stereoisomers that are nonsuperimposable mirror images of one another. A 1:1 mixture of a pair of enantiomers is a racemate or racemic mixture. Diastereomers (or diastereoisomers) are stereoisomers that are not enantiomers, i.e., they are not related as mirror images. If a compound contains a double bond, the substituents can be in the E or Z configuration. Substituents on bivalent cyclic (partially) saturated radicals can have either the cis or trans configuration; for example, if a compound contains a disubstituted cycloalkyl group, the substituents can be in the cis or trans configuration. Therefore, the invention includes enantiomers, diastereoisomers, racemates, E isomers, Z isomers, cis isomers, trans isomers and mixtures thereof, whenever chemically possible. The meaning of all these terms, i.e., enantiomers, diastereoisomers, racemates, E-isomers, Z-isomers, cis-isomers, trans-isomers and mixtures thereof is known to the skilled person. The absolute setting is specified according to the Cahn-Ingold-Prelog system. The configuration on an asymmetric atom is specified by R or S. Resolved stereoisomers whose absolute configuration is not known can be designated by (+) or (-) depending on the direction in which they rotate plane polarized light. For example, resolved enantiomers whose absolute configuration is not known can be designated by (+) or (-) depending on the direction in which plane polarized light rotates.

[00226] When a specific stereoisomer is identified, this means that said stereoisomer is substantially free, i.e. associated with less than 50%, preferably less than 20%, more preferably less than 10%, even more preferably less than 5%, in particular less than 2% and most preferably less than 1% of the other isomers. Thus, when a mGluR2 compound is for example specified as (R), this means that the compound is substantially free of the (S) isomer; when a mGluR2 compound is for example specified as E, this means that the compound is substantially free of the Z-isomer; when a mGluR2 compound is for example specified as cis, this means that the compound is substantially free of the trans isomer.

[00227] Some of the mGluR2 compounds may also exist in their tautomeric form. Such forms, in so far as they may exist, although not explicitly indicated in the above formula, are intended to be included within the scope of the present invention.

[00228] It turns out that a single compound can exist in both stereoisomeric and tautomeric forms.

[00229] For use in medicine, the salts of the compounds of this invention refer to non-toxic "pharmaceutically acceptable salts" (salts of the compounds of the present invention in which the counterion is pharmaceutically acceptable). Other salts may, however, be useful in the preparation or purification of compounds according to this invention or their pharmaceutically acceptable ones, and may encompass acids and bases which are not pharmaceutically acceptable. All salts, whether pharmaceutically acceptable or not, are included within the scope of the present invention.

[00230] The pharmaceutically acceptable acid and base addition salts as mentioned above or hereinafter are intended to comprise the therapeutically active non-toxic acid and base addition salt forms which the compounds of the invention are able to form. Suitable pharmaceutically acceptable salts of the compounds include acid addition salts which may, for example, be formed by mixing a solution of the compound with a solution of a pharmaceutically acceptable acid such as, for example, inorganic acids such as hydrohalic acids, e.g. ., hydrochloric or hydrobromic acid, sulfuric, nitric, phosphoric acid and the like; or organic acids such as, for example, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (i.e. ethanedioic), malonic, succinic (i.e. butanedioic acid), maleic, fumaric, malic, tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic and the like. Conversely, said salt forms can be converted by treatment with an appropriate base into the free base form. Furthermore, where compounds of the invention carry an acid moiety, suitable pharmaceutically acceptable salts thereof can include organic and inorganic bases. Suitable base salt forms include, for example, the ammonium salts, the alkali and alkaline earth metal salts, e.g., the lithium, sodium, potassium, magnesium, calcium and the like salts, salts with organic bases , eg, primary, secondary, and tertiary aliphatic and aromatic amines such as methylamine, ethylamine, propylamine, isopropylamine, the four isomers of butylamine, dimethylamine, diethylamine, diethanolamine, dipropylamine, diisopropylamine, di-n-butylamine , pyrrolidine, piperidine, morpholine, trimethylamine, triethylamine, tripropylamine, quinuclidine, pyridine, quinoline and isoquinoline; the benzathine, N-methyl-D-glucamine, hydrabamine salts, and salts with amino acids such as, for example, arginine, lysine and the like. Conversely, the salt form can be converted by treatment with an acid to the free acid form.

[00231] The term "solvate" comprises the solvent addition forms, as well as their salts, which the compounds of Formula (I) are able to form. Examples of such solvent addition forms are, e.g., hydrates, alcoholates and the like.

Preparation of compounds of Formula (I-B)

[00232] The compounds of Formula (I-B) according to the invention can generally be prepared by a succession of steps, each of which is known to the person skilled in the art. In particular, the compounds can be prepared according to the following synthetic methods.

[00233] The compounds of Formula (I-B) can be synthesized in the form of racemic mixtures of enantiomers that can be separated from each other following art-known resolution procedures. The racemic compounds of Formula (I-B) can be converted into the corresponding diastereomeric salt forms by reaction with a suitable chiral acid. Said diastereomeric salt forms are subsequently separated, for example by selective or fractional crystallization, and the enantiomers are liberated therefrom by alkalis. An alternative way of separating the enantiomeric forms of the compounds of Formula (I-B) involves liquid chromatography or supercritical fluid chromatography (SFC) using a chiral stationary phase. Said pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifically.

A. Preparation of final compounds of Formula (I-B)

[00234] The final compounds according to Formula (IB) can be prepared by reacting an intermediate compound of Formula (11) with a compound of Formula (III) according to reaction scheme (1), a reaction which is carried out under classic Mitsunobu conditions. The reaction is preferably conducted with a phosphine and an azodicarboxylic ester or amide in tetrahydrofuran, 1,4-dioxane, diethyl ether, toluene, benzene, dichloromethane, or mixtures thereof, at -30 to 150 °C under thermal heating or microwave irradiation. The phosphines frequently used are triphenylphosphine and tributylphosphine which are usually combined with dimethyl azodicarboxylate, diethyl azodicarboxylate, diisopropyl azodicarboxylate, di-(4-chlorobenzyl) azodicarboxylate, dibenzyl azodicarboxylate, di-tert azodicarboxylate -butyl, azodicarboxylic acid bis(dimethylamide), azodicarboxylic acid dipiperid, or azodicarboxylic acid dimorpholide. In reaction scheme (1), all variables are as defined in Formula (IB). Reaction Scheme 1

B. Preparation of intermediates Experimental procedure 2

[00235] Intermediate compounds according to Formula (II) can be prepared by subjecting an intermediate of Formula (IV) to conditions which are known to those skilled in the art. This is illustrated in reaction scheme (2) where all variables are defined as mentioned earlier. Methods for achieving these transformations are well known to those skilled in the art. Treatment of the aldehyde of formula (IV) with an organometallic such as methyl lithium bromide or methyl magnesium gives a compound of formula (II). A suitable solvent for this reaction is an ether such as tetrahydrofuran and the reaction is usually carried out at a temperature between -78 °C and 40 °C. In reaction scheme (2), all variables are defined as in Formula (IB). Reaction Scheme 2

Experimental procedure 3

[00236] Intermediate compounds according to Formula (IV) can be prepared by reacting an intermediate of Formula (V) under conditions of dihydroxylation and oxidative cleavage that are known to those skilled in the art and can be carried out, eg with oxone, osmium tetroxide. The process may optionally be carried out in a solvent such as 1,4-dioxane, water and generally at temperatures between about -100°C and about 100°C. A summary of such methods is found in "Comprehensive Organic Transformations", VCH Publishers (1989), RC Larock, pp. 595-596. This is illustrated in reaction scheme (3) where all variables are defined as mentioned earlier. Reaction Scheme 3

Experimental procedure 4

[00237] Intermediate compounds according to Formula (V) can be prepared by coupling reactions, such as Stille or Suzuki reactions of an intermediate of Formula (VI) with a compound of Formula (VII) under conditions that are known to those skilled in the art. The process may optionally be carried out in a solvent such as 1,4-dioxane, water and generally at temperatures between about rt and about 200°C in the presence of a base. This is illustrated in reaction scheme (4) where all variables are defined as mentioned above, where M is trialkyltin, boronic acid or boronate ester, and a palladium and halo catalyst is chlorine, bromine or iodine. reaction 4

Experimental procedure 5

[00238] The intermediate compounds according to Formula (VI) can be prepared according to procedures known in the art by cyclizing an intermediate compound of Formula (VIII) in the presence of a halogenating agent such as for example oxychloride phosphorus (POCl3) (V) in a suitable solvent such as, for example, dichloroethane, stirred under microwave irradiation, for a suitable period of time to allow the completion of the reaction, such as for example 5 min at a temperature between 140 -200°C. In reaction scheme (5), R1 is defined as in Formula (IB) and halo is chlorine, bromine or iodine. Reaction Scheme 5

Experimental procedure 6

[00239] Intermediate compounds according to Formula (VIII) can be prepared by processes known in the art by reacting a hydrazine intermediate of Formula (IX) with acid halides of Formula (X). The reaction may be carried out using an inert solvent, such as for example DCM, in the presence of a base such as for example triethylamine, for example at rt for a suitable period of time to allow completion of the reaction, for example 20 min. . In reaction scheme (6), R1 is defined as in Formula (IB). Reaction Scheme 6

Experimental procedure 7

[00240] Intermediate compounds according to Formula (IX) can be prepared by reacting an intermediate compound of Formula (XI) with hydrazine according to reaction scheme (7), a reaction which is carried out in an inert solvent suitable for the reaction, such as, for example, ethanol, THF or 1,4-dioxane under thermal conditions such as, for example, heating the reaction mixture for example at 160 °C under microwave irradiation for 30 min or thermal heating classic at 70°C for 16 h. In reaction scheme (7), halo is chlorine, bromine, or iodine. Reaction Scheme 7

Experimental procedure 8

[00241] Intermediate compounds according to Formula (XI) can be prepared by reacting an intermediate compound of Formula (XII) with benzyl alcohol according to reaction scheme (8), a reaction which is carried out in a inert solvent suitable for the reaction, such as, for example, N,N-dimethylformamide in the presence of a suitable base, such as for example sodium hydride at rt for a suitable period of time to allow completion of the reaction, such as for example 1 h. In reaction scheme (8), halo is chlorine, bromine or iodine. Reaction scheme 8

Experimental procedure 9

[00242] The intermediate compounds of formula (XII) can be prepared by reacting an intermediate of formula (XIII) with a suitable trifluoromethylating agent, such as for example fluorosulfonyl(difluoro)acetic acid methyl ester, according to the reaction scheme (9). This reaction is carried out in a suitable reaction inert solvent such as, for example, N,N-dimethylformamide in the presence of a suitable coupling agent such as, for example, copper(I) iodide, under thermal conditions. such as, for example, heating the reaction mixture for example at 160°C under microwave irradiation for 45 min. In reaction scheme (9), halo is chlorine, bromine or iodine. Reaction scheme 9

[00243] The starting materials according to Formulas (II), (VII) (X) or (XIII) are compounds that are commercially available or can be prepared according to conventional reaction procedures generally known to those skilled in the art.

[00244] As used herein, the term "composition" is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product that results, directly or indirectly, from combinations of the specified ingredients in the specified amounts.

[00245] As used herein, the term "subject" refers to an animal, preferably a mammal, most preferably a human adult, child or infant, who is or has been the object of treatment, observation or experiment.

[00246] The term "therapeutically effective amount" as used herein means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human that is being sought by a researcher, veterinarian, physician or other clinician, which includes one or more symptoms of the disease or disorder being treated; and/or reducing the severity of one or more symptoms of the disease being treated.

[00247] The combination of compounds (a) SV2A ligand and (b) metabotropic glutamatergic receptor subtype 2 positive allosteric modulator ("PAM") ("mGluR2") or a pharmaceutically acceptable salt or solvate thereof, or metabotropic glutamatergic receptor subtype 2 orthosteric agonist or a pharmaceutically acceptable salt or solvate thereof, whether compounds (a) and (b) are given simultaneously, separately or sequentially, may be beneficial compared to the effect of the compounds ( a) or (b) administered alone. In particular there can be at least one beneficial effect, e.g. a mutual enhancement of the effect of compounds (a) and (b), a more than additive effect, in particular a synergistic effect; additional advantageous effects include, for example, a significantly reduced effective dose for the combination of (a) and (b); an additional therapeutic effect for either compound (a) or (b) alone, a more beneficial side effect profile, or a combined therapeutic effect at an ineffective dosage of one or both of (a) and (b).

[00248] As defined herein, the term "fixed dose ratio of (a) synaptic vesicle protein 2A ligand to (b) compound of Formula (I) of 1:1, calculated based on the ED50 values of the individual compounds (a) and (b)" refers to compositions comprising compounds (a) and (b) in a dose corresponding to 50% of the respective ED50 dose of the individual compounds (a) and (b) or a multiple of this ratio fixed dose. The term " fixed dose ratio of (a) synaptic vesicle protein 2A ligand: (b) compound of Formula (I) of 3:1, calculated based on the ED50 values of the individual compounds (a) and (b) " refers to compositions comprising (b) the compound of Formula (I) in a dose corresponding to 75% of the respective ED50 dose and compound (a) in a dose corresponding to 25% of the respective ED50 dose of the compound (a) or a multiple of this fixed dose ratio, and so on.

[00249] Thus, in another embodiment of the invention, (a) the SV2A ligand and (b) the compound of Formula (I) are present in the pharmaceutical composition in a fixed dose ratio of (a):(b) of about from 1:10 to about 10:1, preferably about 1:5 to about 5:1, more preferably 1:3 to about 3:1, in another embodiment from about 1:1 to about 3: 1; in an alternate 1:3 embodiment; in yet another 1:1 mode; additional 3:1 mode; wherein the fixed dose ratio is calculated based on the ED50 values of the individual compounds (a) and (b).

[00250] Where the present invention is directed to co-therapy or combination therapy, comprising administration of (a) synaptic vesicle protein 2A ligand ("SV2A"); and (b) a PAM/mGluR2 agonist compound, in particular a compound of Formula (I)/(I-A)/(I-B) as defined herein, pharmaceutically or therapeutically effective amount shall mean that amount of the combination of agents taken together as that the combined effect elicits the desired biological or medicinal response. For example, a therapeutically effective amount of cotherapy comprising administration of (a) an SV2A ligand as defined herein (b) a PAM/mGluR2 agonist compound, in particular a compound of Formula (I)/(I-A)/(I-B) as defined herein would be the amount of (a) an SV2A ligand as defined herein and the amount of (b) a PAM/mGluR2 agonist compound, in particular compound of Formula (I)/(I-A)/(I-B) that , when taken together or sequentially have a combined effect that is therapeutically effective. Additionally it will be recognized by one skilled in the art that in the case of co-therapy with a therapeutically effective amount, as in the above example, the amount of the PAM compound/mGluR2 agonist, in particular the compound of Formula (I)/(I-A)/ (I-B), and/or the amount of the appropriate SV2A ligand individually may or may not be therapeutically effective.

[00251] The present invention provides methods of prevention or treatment comprising administering to a subject in need of co-therapy with a therapeutically effective amount of an SV2A ligand and a therapeutically effective amount of a PAM/mGluR2 agonist compound, in particular a compound of the formula (I)/(I-A)/(I-B), as described herein. In order to achieve this objective, the compounds or compositions of this invention must be used in the correct therapeutically effective amount or dose, as described below.

[00252] The optimum dosages and schedules to be administered can be readily determined by those skilled in the art, and will vary with the particular compound used, the mode of administration, the strength of the preparation, the mode of administration, and the advancement of the condition of the illness. Additionally, factors associated with the particular patient being treated, including the patient's age, weight, dietary regimen and time of administration, will result in the need to adjust dosages.

[00253] One skilled in the art will recognize that an effective dosage of the compounds of the present invention can include repeated doses within an extended treatment regimen that will yield clinically significant results.

[00254] The amounts of the PAM compound/mGluR2 agonist, in particular the compound of Formula (I)/(I-A)/(I-B), in the combinations of the invention that are administered on a daily basis can range from about 0.01 to about 2000 mg. Examples of daily amounts of the compound of Formula (I)/(I-A)/(I-B) are 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10, 0, 15.0, 25.0, 50.0, 100, 150, 200, 250, 300, 400, 500, 750 and 1000 milligrams for symptomatic adjustment of dosage to the patient being treated. An effective amount of drug is ordinarily provided at a dosage level of from about 0.01 mg/kg to about 150.0 mg/kg of body weight per day or any range therein. Preferably, the range is from about 0.1 to about 100.0 mg/kg of body weight per day, more preferably, from about 0.5 mg/kg to about 50 mg/kg, most preferably, from about 1.0 to about 25.0 mg/kg of body weight per day. The compounds can be administered in a regimen of 1, 2, 3 or 4 times a day. The amounts of SV2A ligand that are administered on a daily basis can range from about 0.01 to about 7000 mg, preferably it will be between 250 and 5000 mg and most preferably it will be between 500 and 3000 mg. Examples of daily amounts of SV2A ligand are 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 150, 200, 250, 500, 750, 1000, 1500, and 3000 milligrams for the symptomatic adjustment of the dosage of the patient being treated. An effective amount of drug is ordinarily provided at a dosage level of from about 0.01 mg/kg to about 150.0 mg/kg of body weight per day or any range therein. Preferably, the range is from about 0.1 to about 100.0 mg/kg of body weight per day, more preferably, from about 0.5 mg/kg to about 50 mg/kg, most preferably, from about 1.0 to about 25.0 mg/kg of body weight per day. The compounds can be administered in a regimen of 1, 2, 3 or 4 times a day. All amounts mentioned in this and the following paragraphs refer to the free form (i.e., the different salt form). The above values represent free form equivalents, i.e. amounts if the free form were administered. If salts are administered, the amounts need to be calculated as a function of the molecular weight ratio between the salt and the free form.

[00255] The above-mentioned daily doses are calculated for an average body weight of about 70 kg and must be recalculated in the case of pediatric applications, or when used with patients with a substantially deviated body weight.

[00256] Dosages may be presented as one, two, three or four or more sub-doses administered at appropriate intervals throughout the day. The dosage used preferably corresponds to the daily amount of the PAM compound/mGluR2 agonist, in particular the compound of Formula (I)/(I-A)/(I-B), or the SV2A ligand, mentioned above, or a sub-dose thereof, such as 1/2, 1/3, 1/4 of it. A dosage may contain the PAM compound/mGluR2 agonist, in particular the compound (I)/(I-A)/(I-B), or the SV2A ligand or both together, in an amount equal to the ranges or amounts mentioned in the previous paragraphs , for example a dosage form may contain 10 mg, 25 mg, 50 mg, 100 mg, 150 mg, or 200 mg of PAM compound/mGluR2 agonist, in particular compound (I)/(I-A)/(I-B) , 10 mg, 25 mg, 50 mg, 100 mg or 250 mg of SV2A ligand, in separate formulations or in a combined formulation. In one embodiment, the PAM compound/mGluR2 agonist, in particular the compound of Formula (I)/(I-A)/(I-B), is administered once daily (q.d.), in particular as one dose per day, and the ligand of SV2A is administered once or twice daily (q.d. or b.i.d.), in particular as one or two doses per day. In the case where both compounds are to be administered once daily, this can be achieved by administering two separate doses, one with the PAM/mGluR2 agonist compound, in particular the compound of Formula (I)/(I-A)/( I-B), one with the SV2A ligand, or by administering a combination dose containing the PAM/mGluR2 agonist compound, in particular the compound of Formula (I)/(I-A)/(I-B), and SV2A ligand.

[00257] The combinations of the invention can be administered once, twice, three, four or, if desired, multiple times daily. In one embodiment, the combination is administered once daily. In another embodiment, the combination is administered twice daily, or three times daily. Administration of dosages may be in separate dosage forms, i.e., dosage forms containing only PAM/mGluR2 agonist compound, in particular compound of Formula (I)/(I-A)/(I-B), or SV2A ligand only; or by combination dosage forms containing PAM compound/mGluR2 agonist active ingredients, in particular compound of Formula (I)/(I-A)/(I-B), and SV2A ligand. A mixture of combined dosage forms and separate dosage forms can also be used. The dosage forms that can be administered are described hereinafter, oral dosage forms being preferred, in particular tablets or capsules.

[00258] The active ingredients can be formulated into pharmaceutical compositions either separately or as a combined pharmaceutical composition. In the latter case there is provided a pharmaceutical composition comprising a therapeutically effective amount of the PAM/mGluR2 agonist compound, in particular the compound of Formula (I)/(I-A)/(I-B), or a pharmaceutically acceptable salt thereof , and the SV2A ligand, the aforementioned being as specified herein, and a pharmaceutically acceptable carrier.

[00259] In a further aspect, this invention relates to a process for preparing a pharmaceutical composition as specified herein, which comprises intimate mixing of a pharmaceutically acceptable carrier with a therapeutically effective amount of the compound PAM/mGluR2 agonist, in particular compound of Formula (I)/(I-A)/(I-B), or a pharmaceutically acceptable salt or solvate thereof, and a therapeutically effective amount of at least one SV2A binder.

[00260] The combinations provided herein may also be formulated as a combined preparation for simultaneous, separate or sequential use in the prevention or treatment of epilepsy and related disorders; neuropathic pain; migraine or resistant headache; bipolar and related disorders; in neuroprotection; or in the prevention of epileptogenesis. In such a case, the PAM/mGluR2 agonist compound, in particular the compound of Formula (I)/(I-A)/(I-B), is formulated into a pharmaceutical composition containing other pharmaceutically acceptable excipients, and the SV2A ligand it is separately formulated into a pharmaceutical composition containing other pharmaceutically acceptable excipients. Conveniently, these separate pharmaceutical compositions can be part of a kit for simultaneous, separate, or sequential use.

[00261] The individual components of the combination of the present invention can be administered simultaneously or separately at different times during the course of therapy or simultaneously in divided or single combination forms.

[00262] Therefore, the PAM/mGluR2 agonist compounds, in particular the compounds of Formula (I)/(I-A)/(I-B), and the SV2A ligand, individually or in combination, can be formulated into various pharmaceutical compositions suitable for administration purposes. In these, a therapeutically effective amount of the particular compound, or both compounds, is combined with a pharmaceutically acceptable carrier, which carrier can take a wide variety of forms depending on the form of preparation desired for administration. The pharmaceutical compositions can be prepared as medicaments to be administered orally, parenterally (including subcutaneously (s.c.), intramuscularly (i.m.), and intravenously (i.v.)), rectally, transdermally, buccally, or nasally. Pharmaceutical compositions can also be prepared to be administered directly to the nervous system by routes including, but limited to, intracerebral, intraventricular, intracerebroventricular, intrathecal, intracisternal, intraspinal and/or perispinal routes by administration through intracranial or intravertebral needles and/or catheters with or without pump devices. Compositions suitable for oral administration include powders, granules, pellets, tablets, compressed or coated pills, dragées, sachets, hard or gelatin capsules, syrups and suspensions. Compositions suitable for parenteral administration include aqueous or non-aqueous solutions or emulsions, while for rectal administration compositions suitable for administration include suppositories with a hydrophilic or hydrophobic vehicle. For topical administration transdermal delivery systems may be used and for nasal administration suitable aerosol delivery systems may be used.

[00263] For example, in preparing the compositions for oral administration, any of the usual pharmaceutical media may be employed such as, for example, water, glycols, oils, alcohols and the like in the case of oral liquid compositions such as suspensions, syrups, elixirs, emulsions and solutions; or solid carriers such as starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like in the case of solid compositions. For parenteral compositions, the carrier will usually comprise sterile water, at least in large part, although other ingredients such as solubilizers, emulsifiers or additional auxiliaries may be added thereto. Injectable solutions can be prepared in which the carrier comprises saline solution, glucose solution or a mixture of both. Injectable suspensions may also be prepared, in which case appropriate liquid carriers, suspending agents and the like may be employed. Also included are solid form preparations intended to be converted, shortly before use, to liquid form preparations such as powders for reconstitution. In compositions suitable for percutaneous administration, the carrier optionally comprises a skin penetration enhancing agent and/or a wetting agent, optionally combined with suitable skin compatible additives in minor proportions. The PAM compound/mGluR2 agonist, in particular the compound of Formula (I)/(I-A)/(I-B), or SV2A ligand, or combinations thereof, can also be administered via oral inhalation or insufflation by suitable formulations for this type of administration such as a solution, a suspension or a dry powder. Pharmaceutical compositions suitable for administration in the form of aerosols or sprays are, for example, suspensions of the compound PAM/mGluR2 agonist, in particular the compound of Formula (I)/(I-A)/(I-B), or SV2A ligand, or both , in a pharmaceutically acceptable liquid carrier, such as ethanol or water, or a mixture thereof. If required, the formulation can also additionally contain other pharmaceutical auxiliaries such as surfactants, emulsifiers and stabilizers as well as a propellant. Such a preparation usually contains the active compound in a concentration of approximately 0.1 to 50%, in particular approximately 0.3 to 3%, by weight.

[00264] The pharmaceutical compositions may contain the compound active ingredient PAM/mGluR2 agonist, in particular compound of Formula (I)/(I-A)/(I-B), or the SV2A ligand, or both combined in a concentration of about 0.1% to about 50%, or about 1% to about 30%, or about 3% to about 20%, or about 5% to about 20%, all percentages being by weight, wherein the total of all components in said pharmaceutical compositions does not exceed 100%. In pharmaceutical compositions containing both the PAM compound/mGluR2 agonist compound, in particular the compound of Formula (I)/(I-A)/(I-B), or the SV2A ligand, the PAM compound/mGluR2 agonist, in particular the compound of the Formula (I)/(I-A)/(I-B), is present in a concentration of about 0.1% to about 50%, or about 1% to about 30%, or about 3% to about 20%, or about 5% to about 20%; and the SV2A ligand is present at a concentration of about 3% to about 50%, or about 5% to about 50%, or about 10% to about 50%, or about 10% to about from 40%, or from about 10% to about 30%, wherein the total of all components in said pharmaceutical compositions does not exceed 100%.

[00265] Pharmaceutical compositions may conveniently be presented in unit dosage form for ease of administration and uniformity of dosage. Examples include tablets (including scored or coated tablets), capsules, pills, suppositories, powder packets, wafers, injectable solutions or suspensions and the like, and segregated multiples thereof. Of interest are solid dosage forms for oral administration such as tablets or capsules.

[00266] The solid dosage forms in unit dosage form may be packaged in any known package, blister packs being preferred, in particular for tablets and capsules. Where the PAM compound/mGluR2 agonist, in particular the compound of Formula (I)/(I-A)/(I-B), and SV2A ligand are formulated separately they may be packaged in separate blisters, but a blister could equally well comprise dosage forms PAM compound/mGluR2 agonist unit, in particular the compound of Formula (I)/(I-A)/(I-B), and the SV2A ligand, for example a row of PAM compound/mGluR2 agonist units, in particular compound of Formula (I)/(I-A)/(I-B), and another with SV2A ligand. Other possibilities may also be possible.

[00267] The combinations of this invention can be used to treat or prevent epilepsy and related disorders; neuropathic pain; migraine or resistant headache; and bipolar and related disorders; or they can be used as a neuroprotector or to prevent epileptogenesis.

[00268] As used herein, the term "treatment" is intended to refer to all processes, in which there may be a slowdown, interruption, arrest or termination of the progression of a disease or a relief of symptoms, but does not necessarily indicate a total elimination of all symptoms.

[00269] As used herein, unless otherwise noted, the terms "epilepsy and related disorders" or "epilepsy or related disorders" shall mean any disorder in which an individual (preferably an adult, child or human infant) experiences - experiences one or more seizures and/or tremors. Suitable examples include, but are not limited to, epilepsy (including, but not limited to, location-related epilepsy, generalized epilepsies, epilepsies with both generalized and local seizures, and the like), partial-onset seizures with or without generalization, myoclonic seizures, primary generalized tonic-clonic seizures in particular in patients with idiopathic generalized epilepsy, seizures associated with Lennox-Gastaut syndrome, seizures as a complication of a disease or condition (such as seizures associated with encephalopathy, phenylketonuria, juvenile Gaucher, progressive myoclonic epilepsy of Lundborg, stroke, head injury, stress, hormonal changes, drug use or withdrawal, alcohol use or withdrawal, sleep deprivation, fever, infection, and the like), status epilepticus (convulsive or not convulsive), essential tremor, restless limb syndrome, and the like. Preferably, the disorder is selected from epilepsy (regardless of type, underlying cause, or origin), essential tremor, or restless limb syndrome. More preferably, the disorder is epilepsy (regardless of type, underlying cause or origin) or essential tremor. A particular example of epilepsy is refractory epilepsy, also referred to as treatment- or therapy-resistant epilepsy. This term is often used when patients have failed three or more antiepileptic drugs (AEDs). Refractory epilepsy also includes partial refractory epilepsy and refractory generalized (including idiopathic or symptomatic) epilepsy.

[00270] As used herein, the term "neuropathic pain" includes pain resulting from chronic or debilitating conditions or disorders. Chronic or debilitating conditions or disorders that can lead to neuropathic pain include, but are not limited to, painful diabetic peripheral neuropathy, postherpetic neuralgia, trigeminal neuralgia, poststroke pain, pain associated with multiple sclerosis, pain associated with neuropathies such as idiopathic or post-traumatic neuropathy and mononeuritis, neuropathic pain associated with HIV, neuropathic pain associated with cancer, neuropathic pain associated with carpal tunnel, pain associated with spinal cord injury, low back and neck pain , reflex sympathetic dystrophy, phantom limb syndrome, and other pain syndromes associated with chronic and debilitating conditions.

[00271] As used herein, the term "migraine" shall mean a chronic, episodic and debilitating condition that is diagnosed by the presence of moderate to severe pulsating headaches lasting between 4 and 72 h, which includes migraine without aura and migraine with aura . As used herein, "migraine without aura" shall mean at least five attacks fulfilling the following criteria: (a) the headache attack lasts 4 and 72 hours without the headache having at least two of the following characteristics: unilateral location, pulsating quality, moderate to severe intensity with direct influence on activities of daily living, and aggravation by climbing stairs or similar routines; and (b) during the headache, at least one of the following occurs: nausea and/or vomiting, and photophobia and phonophobia. As used herein, "migraine with aura" shall mean at least two attacks accompanied by at least 3 of the following 4 features: (a) one or more fully reversible aura symptoms: (b) at least one gradually developing aura symptom over more than 4 minutes or two or more symptoms occurring in succession; (c) no aura symptoms lasting more than 60 minutes; (d) a headache occurs before, simultaneously with, or after the aura, with a free interval between aura and headache of less than about 60 minutes.

[00272] As used herein, the term "bipolar and related disorders" shall include bipolar I disorder (eg, single manic episode, most recent hypomanic episode, most recent manic episode, most recent mixed episode, most recent depressive episode, and most recent unspecified episode), Bipolar II Disorder, Cyclothymic Disorder, and Bipolar Disorder Not Otherwise Specified (as these terms are defined by their diagnostic criteria, in the Diagnostic and Statistical manual of Mental Disorders 4th Edition, Text Review, American Psychiatric Association, 2000 (DSM-IV-TR) or the 5th Edition, Text Revision, American Psychiatric Association, 2013 (DSM-5TM)). Rather, bipolar disorder is characterized by depressive and manic (or hypomanic) phases, where the phases cycle. Preferably, the bipolar disorder is bipolar I disorder or bipolar II disorder. As used herein, "mania" shall include mania or a manic mood phase, regardless of the underlying cause. As used herein, the term "bipolar mania" is intended to mean mania associated with, characteristic of, or symptomatic of a bipolar disorder. Thus, methods of treating bipolar mania of the present invention are directed to methods of treating the mania and/or manic phase of bipolar disorders. As used herein, the term "bipolar depression" is intended to mean depression associated with, characteristic of, or symptomatic of a bipolar disorder. Thus, methods of treating bipolar depression of the present invention are directed to methods of treating the depression and/or depressive phase of bipolar disorders. As used herein, unless otherwise noted, the terms "in cycle" or "bipolar cycle" shall refer to the shift in mood between depressive and manic phases characteristic of bipolar disorders. Thus, the present invention includes methods for stabilizing said cycle, including, but not limited to, decreasing cycle frequency and/or decreasing the magnitude of manic and/or depressive phases.

[00273] Thus, in one embodiment, the pharmaceutical composition of the present invention can be used for mood stabilization, in particular mood stabilization for manic depression.

[00274] As used, the term "epileptogenesis" refers to the gradual process by which epilepsy develops. This process can occur after insults to the brain or a variety of conditions, including neurodegenerative diseases, traumatic brain injury, stroke, brain tumor, central nervous system infections, and status epilepticus; or it may occur after gene mutations.

[00275] As used herein, the term "anxiety" refers in particular to generalized anxiety disorder.

[00276] As used herein, the term "about" has its conventional meaning. In particular embodiments, when in relation to a numerical value, it can be interpreted as meaning the numerical value ± 10%, or ± 5%, or ± 2%, or ± 1%, or ± 0.5%, or ± 0.1%. In other embodiments, the precise value is intended, i.e., leaving aside the words "about".

[00277] "And/or" means that each or both or all of the components or characteristics of a list are possible variants, especially two or more of them in an alternative or cumulative manner.

[00278] As used herein, the terms "a", "an", "the/a" and similar terms used in the context of the present invention (especially in the context of the claims) are to be interpreted to encompass both singular and plural unless otherwise indicated herein or clearly contradicted by the context.

EXAMPLES

[00279] The following Examples are presented to assist in understanding the invention, and are not intended and should not be considered as limiting in any way the invention presented in the claims that follow subsequently.

A) COMPOUNDS OF THE FORMULA (I-B) - CHEMISTRY AND IN VITRO TEST

[00280] Various methods for preparing the compounds of Formula (I-B) of this invention are illustrated in the following Examples. Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification.

[00281] Henceforth, "aq." means watery; "DCE" means 1,2-dichloroethane, "DCM" means dichloromethane; "DIPE" means diisopropylether; "DIPEA" means N,N-diisopropylethylamine; "DMF" means N,N-dimethylformamide; "ES" means electrospray; "Et3N" means triethylamine; "Et2O" means diethyl ether; "EtOAc" means ethyl acetate; "h" means hours; "HPLC" means high performance liquid chromatography; "HRMS" means high resolution mass spectrometry/spectrometry; "l" or "L" means liter; "LRMS" means low resolution mass spectrometry/spectrometry; "MeOH" means methanol; "min" means minute(s); "mp" means melting point; "Pd(PPh3)4" means tetrakis(triphenylphosphine)palladium(0); "RP" means reverse phase; "r.t." means ambient temperature; "s" means seconds; "sat." means saturated; "SFC" means supercritical fluid chromatography; "Sun." means solution; "THF" means tetrahydrofuran.

[00282] The microwave-assisted reactions were carried out in a single-mode reactor: Sixty EXP InitiatorTM microwave reactor (Biotage AB), or in a multimodal reactor: MicroSYNTH Labstation (Milestone, Inc.).

[00283] Thin layer chromatography (TLC) was carried out on silica gel 60 F254 plates (Merck) using reagent grade solvents. Open column chromatography was performed on silica gel, particle size 60 Å, mesh = 230-400 (Merck) using standard techniques. Automated rapid column chromatography was performed using Merck ready-to-connect cartridges, irregular silica gel, particle size 15-40 μm (disposable normal phase rapid columns) on an Armen Instrument SPOT or LRÁPIDA system.

[00284] The absolute stereochemical configuration for some of the compounds was determined using vibrational circular dichroism (VCD). They were measured in a Bruker Equinox 55 spectrometer equipped with a PMA 37, in a KBr liquid cell using CD2Cl2 as solvent (PEM: 1350 cm-1, LIA: 1mV, resolution: 4 cm-1). A description of the use of VCD for absolute configuration determination can be found in Dyatkin A.B. et al., Chirality, 14: 215-219 (2002 ).

[00285] Whenever the notation "RS" is indicated here it denotes that the compound is a racemic mixture, unless otherwise indicated. The stereochemical configuration for some compounds was designated "R" or "S" when the mixture was separated; for some compounds, the stereochemical configuration has been designated as "*R" or "*S" when the absolute stereochemistry is indeterminate even though the compound itself has been isolated as a single stereoisomer and is enantiomerically pure. The enantiomeric excess of the compounds reported here was determined by analysis of the racemic mixture by supercritical fluid chromatography (SFC), followed by SFC comparison of the separated enantiomer(s). Preparation of Intermediates Description 1 - Intermediate 1

[00286] Cyclopropylacetic acid ([CAS 5239-82-7], 50 g, 500 mmol) was dissolved in CH2Cl2 (300 mL), then SOCl2 (100 mL) was added. The reaction mixture was stirred at 60 °C for 2 h and then the solvent was evaporated to give intermediate 1 (53 g, 90%) which was used without further purification. Description 2 - Intermediate 2

[00287] To a solution of 2,4-dichloro-3-iodopyridine ([CAS 34378136-2], 290 g, 1058 mmol) in DMF (1.7 L) was added 2,2-difluoro-2-(fluorosulfonyl )methyl acetate ([CAS 680-15-9], 403 g, 2098 mmol) and Cul (403 g, 2.13 mol), the reaction was then heated at 100 °C for 5 h.

[00288] The reaction was cooled and filtered. The filtrate was diluted with H2O and extracted with Et2O and washed with an NH3 solution. The organic layer was dried (Na2SO4), filtered and concentrated in vacuo to give intermediate 2 (160g) which was used without further purification. Description 3 - Intermediate 3

[00289] To a solution of NaH (60% in oil, 24 g, 600 mmol) in DMF (2 L) at 0 °C was added benzyl alcohol (35 g, 325 mmol), then reaction was stirred for 2 min . Intermediate 2 (160 mg, 741 mmol) was added in one portion, and stirred at 0 °C for 1 h. The reaction was diluted by the addition of H2O and extracted with Et2O. The organic layer was dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel (eluent: petroleum ether/EtOAc = 20/1). The pure fractions were collected and the solvent was evaporated to give Intermediate 3 (100g, 38%). Description 4 - Intermediate 4

[00290] To a solution of Intermediate 3 (100 g, 277 mmol) in 1,4-dioxane (1.5 L) was added NH2NH2 hydrate (85% solution in water, 300 g, 9.11 mol), the reaction was then heated in a sealed tube at 160 °C for 2 h. The mixture was concentrated in vacuo, dissolved in DCM, washed with NaHCO 3 . The organic layer was dried (Na2SO4), filtered and concentrated in vacuo to give Intermediate 4 (90g, 90%) which was used without further purification. Description 5 - Intermediate 5

[00291] To a solution of intermediate 4 (90 g, 318 mmol) in CH2Cl2 (1.5 L) was added triethylamine (64.3 g, 636 mmol), the mixture was cooled to 0 °C, then a solution of Intermediate 1 (53 g, 449 mmol) in CH2Cl2. The solution was stirred at RT for 1 h. The reaction mixture was washed with sat. sat. here of NaHCO3, and extracted with CH2Cl2. The organic layer was dried (Na2SO4), filtered and concentrated in vacuo to give intermediate 5 (104.4 g, 90%).

[00292] The following intermediates were synthesized following a synthetic sequence analogous to that reported in Description 5 (D5). Intermediate Acid chloride Conditions Description 6 (a) Intermediate 9

[00293] To a solution of intermediate 5 (101 g, 277 mmol) in CH3CN (1.2 L) were added phosphorus (V) oxychloride (84.7 g, 553 mmol) and N,N-diisopropylethylamine ( 71.3 g, 553 mmol). The reaction mixture was stirred at 90 °C for 38 h. The reaction was then diluted with DCM and washed with Na2CO3 solution. The organic layer was dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel (eluent: petroleum ether/EtOAc = 4/1). The pure fractions were collected and the solvent was evaporated to give intermediate 9 (31.39 g, 41%).(b) Intermediate 10

[00294] The reaction was performed in 4 batches, then combined for processing and purification. To a solution of intermediate 6 (7g, 20.6mmol) in DCE (50ml) was added N,N-diisopropylethylamine (3.96ml, 22.69mmol) and then phosphorus oxychloride (2.12 mL, 22.69 mmol) and the reaction mixture was heated in a microwave at 150 °C for 5 min. Then DCM was added and the organic layer was washed with sat. sat. of NaHCO3, dried (Na2SO4), and concentrated in vacuo to give the desired compound, which was purified by column chromatography (elution gradient: 100% DCM to 2% MeOH.NH3 in DCM) to give the intermediate 10 (2.5g, 49%).

[00295] The following intermediates were synthesized following a synthetic sequence analogous to that reported in Description 6(a) or (b). Description 7 - Intermediate 13

[00296] (Ph3P)4Pd (2.096 g, 1.81 mmol) was added to a stirred solution of intermediate 9 (10 g, 36.28 mmol) and 4,4,5,5-tetramethyl-2-vinyl-1 ,3,2-dioxoborolane ([CAS 75927-49-0], 7.77 mL, 43.53 mmol) in deoxygenated dioxane (30 mL) and a saturated solution of deoxygenated NaHCO 3 (30 mL) under nitrogen. The mixture was stirred at 100 °C for 18 h. The mixture was diluted with EtOAc/water and filtered through a pad of diatomaceous earth. The filtrate was treated with brine and extracted with EtOAc. The organic layer was separated, dried (Na2SO4), filtered and the solvents evaporated in vacuo. The crude product was purified by flash column chromatography (silica; EtOAc in CH 2 Cl 2 0/100 to 5/95). The desired fractions were collected and concentrated in vacuo to give intermediate 13 (6.08 g, 63%) as a yellow solid.

[00297] The following intermediates were synthesized following a synthetic sequence analogous to that reported in Description 7. Description 8 (a) Intermediate 17

[00298] Osmium tetraoxide (2.5% in t-BuOH, 10.103 mL, 0.781 mmol) and then sodium periodate (12.53 g, 58.58 mmol) in water (48.5 mL) were added to a suspension of Intermediate 13 (6.08g, 20.02mmol) in dioxane (192ml). The mixture was stirred at room temperature for 2 h.

[00299] The mixture was treated with water and EtOAc and filtered off through a pad of diatomaceous earth. The filtrate was extracted with EtOAc. The organic layer was separated, dried (Na2SO4), filtered and the solvents evaporated in vacuo. The crude product was washed with Et2O and filtered and dried to give intermediate 17 (4.25 g, 79%) as a brown solid. (b) Intermediate 18

[00300] A suspension of sodium periodate (5.04 g, 23.54 mmol) in distilled water (19 mL) was added to a stirred solution of osmium tetraoxide (2.5% in t-BuOH, 4.06 mL, 0.31 mmol) and intermediate 14 (2.08 g, 7.85 mmol) in dioxane (75 mL). The mixture was stirred at room temperature for 150 min, then the mixture was treated with sat NaHCO 3 and brine, and extracted with DCM. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The product was triturated with Et2O and filtered in vacuo, and finally placed in a desiccator at 50 °C for 18 h, to give intermediate 18 (1.6 g, 80%) as a brown solid.

[00301] The following intermediates were synthesized following a synthetic sequence analogous to that reported in Description 8. Description 9 (a) Intermediates 21a, 21b and 21c

[00302] Methylmagnesium bromide (1.4 M in THF, 12.40 ml, 17.37 mmol) was added dropwise to a stirred suspension of intermediate 17 (4.25 g, 15.79 mmol) in THF (281.07 mL) at -20°C under N2 atmosphere. The mixture was stirred at -20°C for 45 minutes. The crude was treated with a sol. sat. of NH4Cl and extracted with EtOAc. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica; MeOH in DCM 0/100 to 4/96). The desired fractions were collected and concentrated in vacuo to give intermediate 21a (racemic mixture) (2.96 g, 66%). Intermediate 21a (1.82 g) was purified by chiral SFC: [Stationary phase: CHIRALPAK AD-H (5 µm 250 x 20 mm), Mobile phase: 80% CO2, 20% EtOH] yielding 21b (enan- R-thiomer) (0.453 g, 10%) as a pale gray solid and Intermediate 21c (S-enantiomer) (0.439 g, 10%).(b) Intermediate 22

[00303] Methylmagnesium bromide (1.4 M in THF, 3.97 mL, 5.56 mmol) was added dropwise to a stirred suspension of intermediate 18 (1.23 g, 5.06 mmol) in THF ( 90 mL) at -20°C under N2 atmosphere. The mixture was stirred at -20°C for 45 minutes. The crude was treated with a sol. sat. of NH4Cl and extracted with EtOAc. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica; MeOH in DCM 0/100 to 4/96). The desired fractions were collected and concentrated in vacuo. The residue thus obtained was triturated with Et2O to give intermediate 22 (620 mg, 35%) as a pale yellow solid.

[00304] The following intermediates were synthesized following a synthetic sequence analogous to that reported in Description 9.

[00305] Intermediate 24a has been further separated into Intermediate 24b and Intermediate 24c: Preparation of Final Compounds of Formula (IB) Example 1 (a) Synthesis of Compounds 4-b, 6-b and 5-b

[00306] DIAD (2.07 mL, 10.52 mmol) was added dropwise to a stirred solution of intermediate 21a (2 g, 7.01 mmol), 2,4-difluorophenol (1.00 mL, 10, 52 mmol) and triphenylphosphine (2.76 g, 10.52 mmol) in THF (74.18 mL) at 0 °C and under a nitrogen atmosphere. The mixture was stirred at 100°C for 10 minutes under microwave irradiation. The mixture was diluted with EtOAc and washed with sat. sat. of NaHCO3. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica; MeOH in DCM 0/100 to 97/3). The desired fractions were collected and concentrated in vacuo. The residue was triturated with DIPE to give compound 4-b (1.46 g, 52%) as a white solid, which was purified by chiral SFC [Stationary phase: Chiralpak AD (5 µm 250 * 30 mm, Mobile phase: 85% CO2, 15% iPrOH], yielding compound 6-b (0.659 g, 24%) and compound 5-b (0.693 g, 25%).(b) Alternative synthesis of compound 6-b

[00307] DIAD (31.06 μL, 0.16 mmol) was added dropwise to a stirred solution of intermediate 21b (30 mg, 0.11 mmol), 2,4-difluorophenol (15.07 μL, 0. 16 mmol) and triphenylphosphine (41.38 mg, 0.16 mmol) in THF (1.11 mL) at 0 °C and under a nitrogen atmosphere. The mixture was stirred at 100°C for 10 minutes under microwave irradiation. The mixture was diluted with EtOAc and washed with sat. sat. of NaHCO3. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica; MeOH in DCM 0/100 to 97/3). The desired fractions were collected and concentrated in vacuo. The residue was triturated with DIPE to give compound 6-b (40 mg, 96%) as a white solid. (c) Synthesis of compound 6-b hydrochloride salt (.HCl)

[00308] DIAD (207.06 μL, 1.05 mmol) was added dropwise to a stirred solution of intermediate 21b (200 mg, 0.70 mmol), 2,4-difluorophenol (100.45 μL, 1, 05 mmol) and triphenylphosphine (275.84 mg, 1.0516 mmol) in THF (4 mL) at 0°C and under a nitrogen atmosphere. The mixture was stirred at 100°C for 15 minutes under microwave irradiation. The mixture was diluted with EtOAc and washed with sat. sat. of NaHCO3. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by RP HPLC (Stationary phase: C18 XBridge 30 x 100 mm 5 μm, Mobile phase: 60% and 0.1% NH4CO3H/NH4OH solution gradient pH 9 in Water, 40% CH3CN solution at 43% and 0.1% NH 4 CO 3 H/NH 4 OH pH 9, 57% CH 3 CN, yielding a white solid residue which was dissolved in Et 2 O (8 mL) and 1,4-dioxane (0.5 mL). To the solution thus obtained, HCl (4 M in dioxane, 200 µL) was added dropwise. The precipitated white solid was filtered, washed with Et2O, dried (Na2SO4) and evaporated in vacuo. The white residue thus obtained was triturated with Et2O to give compound 6-b.HCl (110 mg, 36%) as a white solid.

[00309] The following compounds were synthesized following a synthetic sequence analogous to that reported in Example 1(b), starting from intermediate 21b. Example 2 Synthesis of compound 7-b

[00310] Procedure (a): DIAD (31.06 µL, 0.158 mmol) was added dropwise to a stirred solution of intermediate 21b (30 mg, 0.105 mmol), 3,5-difluorophenol (20.52 mg, 0.158 mmol) and triphenylphosphine (41.38 mg, 0.158 mmol) in THF (1.113 mL) at 0 °C and under a nitrogen atmosphere. The mixture was stirred at 100°C for 10 minutes under microwave irradiation. The mixture was diluted with EtOAc and washed with sat. sat. of NaHCO3. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica; MeOH in DCM 0/100 to 96/4). The desired fractions were collected and concentrated in vacuo. The residue was triturated with DIPE to give compound 7-b (21 mg, 50%) as a white solid.

[00311] Procedure (b): Alternatively, compound 7 was also synthesized following a synthetic sequence analogous to that reported in Example 1(b), starting from intermediate 21b. Example 3 Synthesis of compound 8-b

[00312] Procedure (a): DIAD (31.06 μL, 0.158 mmol) was added dropwise to a stirred solution of intermediate 21b (30 mg, 0.105 mmol), 3,4-difluorophenol (20.52 mg, 0.158 mmol) and triphenylphosphine (41.38 mg, 0.158 mmol) in THF (1.11 mL) at 0 °C and under a nitrogen atmosphere. The mixture was stirred at 100°C for 10 minutes under microwave irradiation. The mixture was diluted with EtOAc and washed with sat. sat. of NaHCO3. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo. The residue was purified by flash column chromatography (silica; MeOH in DCM 0/100 to 96/4). The desired fractions were collected and concentrated in vacuo. The residue was triturated with DIPE to give compound 8-b (10.6 mg, 25%) as a white solid.

[00313] Procedure (b): Alternatively, compound 8-b was also synthesized following a synthetic sequence analogous to that reported in Example 1(b), starting from intermediate 21b. Example 4 Synthesis of compound 15-b

[00314] Procedure (a): DIAD (155.3 μL, 0.789 mmol) was added dropwise to a stirred solution of intermediate 21b (150 mg, 0.526 mmol), 2,4,6-trifluorophenol (116, 8 mg, 0.789 mol) and triphenylphosphine (206.88 mg, 0.789 mmol) in THF (5.56 mL) at 0°C and under nitrogen atmosphere. The mixture was stirred at 100°C for 10 minutes under microwave. The mixture was diluted with DCM and washed with sat. NaHCO3 sat. The organic layer was separated, dried (Na2SO4), filtered and concentrated in vacuo, then purified by flash column chromatography (silica; 7N MeOH/NH3 in DCM 0/100 to 90/10). The desired fractions were collected and concentrated in vacuo. O was purified by RP HPLC (Stationary phase: C18 XBridge 30 x 100 mm 5 μm, Mobile phase: 54% and 0.1% NH4CO3H/NH4OH solution gradient pH 9 in Water, 46% CH3CN solution to NH4CO3H /64% and 0.1% NH 4 OH pH 9, 36% CH 3 CN) yielding a colorless oil which crystallized upon standing (2 days). The solid was triturated with heptane to give compound 15-b (129.8 mg, 59%) as a white solid.

[00315] Procedure (b): Alternatively, compound 15-b was also synthesized following a synthetic sequence analogous to that reported in Example 1(b), starting from intermediate 21b. Example 5 Synthesis of compounds 1-b, 2- be 3-b

[00316] Compounds 1-b, 2-b and 3-b were synthesized following the procedure described in Example 1(a). Thus, the reaction of DIAD (500.05 µL, 2.54 mmol), intermediate 21a (483 mg, 1.69 mmol), 4-fluorophenol (227.77 mg, 2.03 mmol) and triphenylphosphine (666.14 mg, 2.54 mmol) in THF (17.91 mL) as described in Example 1(a) gave a residue which was purified by flash column chromatography (silica; EtOAc in DCM 0/100 to 90/10). The desired fractions were collected and concentrated in vacuo. The resulting residue was triturated with DIPE to give compound 1-b (320 mg, 50%) as a white solid, which was purified by chiral SFC [Stationary phase: Chiralpak AD (5 µm 250 * 30 mm, Mobile phase: CO2 77%, 23% MeOH], yielding compound 2-b (131 mg, 20%) and compound 3-b (129 mg, 20%) as white solids. Example 6 Synthesis of compounds 24-b, 26 -b, and 27-b

[00317] Compounds 24-b, 26-b and 27-b were synthesized following the procedure described in Example 1(a). Thus, the reaction of DIAD (364.57 µL, 1.85 mmol), intermediate 22 (320 mg, 1.23 mmol), 2,4-difluorophenol (176.86 µL, 1.85 mmol) and triphenylphosphine (485 .67 mg, 1.85 mmol) in THF (13.06 mL) as described in Example 1(a) gave a residue which was purified by flash column chromatography (silica; MeOH in DCM 0/100 to 96/4) . The desired fractions were collected and concentrated in vacuo to give a colorless oil which crystallized with DIPE to give compound 24 as a white solid, which was purified by RP HPLC (Stationary phase: C18 XBridge 30 x 100 mm 5 µm, Mobile phase: Gradient from 54% 0.1% NH4CO3H/NH4OH pH 9 solution in Water, 46% CH3CN solution to 64% 0.1% NH4CO3H/0.1% NH4OH pH 9, 36% CH3CN solution giving a colorless oil which was crystallized after trituration with heptane to give 240 mg (52%) of compound 24-b as a white solid, which was further purified by chiral SFC (Stationary phase: CHIRALPAK AD-H 5 µm 250 x 20 mm; mobile phase: 85% CO 2 , 15% iPrOH (0.3% iPrNH 2 )], yielding compound 26-b (103 g, 22%) and compound 27-b (107 g, 23%).

[00318] The following compounds were obtained following a synthetic sequence similar to that reported in Example 1(a).

[00319] The following compounds were synthesized following a synthetic sequence as reported in Example 1(b), starting from the indicated intermediates.

[00320] Table A below lists additional compounds of Formula (IB) that were prepared by analogy with the above examples (Exp. no.). Table A: Exemplary compounds according to Formula (IB).# indicates that the experimental procedure is described in the examples.

analytical part Optical Rotations

[00321] Optical rotations were measured on a PerkinElmer 341 polarimeter with a sodium lamp and reported as follows: [α]° (À, c g/100 mL, solvent, T°C).

[00322] [α]xT = (100α) / (l x c) : where l is the path length in dm and c is the concentration in g/100 mL for a sample at a temperature T (°C) and a wavelength At (in nm). If the wavelength of light used is 589 nm (the sodium D line), then the symbol D could be used instead. The sign of rotation (+ or -) must always be given. When using this equation, the concentration and solvent are always given in parentheses after rotation. Rotation is reported using degrees and no units of concentration are given (assumed to be g/100 mL).

LCMS

[00323] For characterization by (LC)MS of the compounds of the present invention the following methods were used.

general procedure

[00324] The measurement by High Performance Liquid Chromatography (HPLC) was performed using an LC pump, a diode array (DAD) or a UV detector and a column as specified in the respective methods. If necessary, additional detectors were included (see methods table below).

[00325] The column flow was conducted to the Mass Spectrometer (MS) which was configured with an ion source at atmospheric pressure. It is within the knowledge of the skilled person to adjust the tuning parameters (eg screening range, residence time...) in order to obtain ions allowing the identification of the nominal monoisotropic molecular weight (MW) of the compound. Data acquisition was performed with appropriate software.

[00326] Compounds are described by their experimental retention times (Rt) and ions. If not specified otherwise in the data table, the reported molecular ion corresponds to [M+H]+ (protonated molecule) and/or [M-H]- (deprotonated molecule). In case the compound is not directly ionizable, the type of adduct is specified (i.e., [M+NH4]+, [M+HCOO]-, etc...). For molecules with multiple isotopic patterns (Br, Cl..), the reported value is that obtained for the lowest isotopic mass. All results were obtained with experimental uncertainties that are commonly associated with the method used.

[00327] Henceforth, "SQD" means Single Quadrupole Detector, "RT" room temperature, "BEH" bridged ethylsiloxane/silica hybrid, "HSS" High Strength Silica, "DAD" Diode Array Detector.Table B : LCMS Method Codes (Flow expressed in mL/min; column temperature (T) in °C; Run time in minutes).

melting points

[00328] The values are peak values, and are obtained with experimental uncertainties that are commonly associated with this analytical method.

Mettler FP 81HT/ FP90 device

[00329] For a number of compounds, melting points were determined in open capillary tubes in an FP 81HT/FP90 device (Mettler-Toledo). Melting points were measured with a temperature gradient of 1, 3, 5 or 10°C/minute. The maximum temperature was 300°C. The melting point was read from a digital monitor.Table C: Physical-chemical data for some compounds, retention time (Rt) in min, [M+H]+ peak (protonated molecule), LCMS method and mp ( melting point in °C). (nd = not determined).

SFC-MS general procedure

[00330] The SFC measurement was performed using a Berger Instruments Analytical System comprising an FCM-1200 dual pump fluid control module for administration of carbon dioxide (CO2) and modifier, an automatic liquid sampling device from CTC Analytics, a TCM-20000 thermal control module for heating the column from room temperature to 80°C. An Agilent UV 1100 photodiode array detector equipped with a high pressure flow cell supporting up to 400 bars was used. The column flow was separated into an MS spectrometer. The MS detector was configured with an atmospheric pressure ionization source. The following ionization parameters for the Waters ZQ mass spectrometer are: head top: 9 μa, source temperature: 140°C, cone: 30 V, probe temp 450°C, extractor 3 V, desolvation gas 400 L/hr, cone gas 70 L/hr. Nitrogen was used as the nebulizer gas. Data acquisition was performed with a Waters-Micromass MassLynx-Openlynx data system.

[00331] Method 1: In addition to the general procedure: The analytical chiral separation in SFC-MS was carried out on a CHIRALPAK AD DAICEL column (10 μm, 4.6 x 250 mm) at 35°C with a flow rate of 3, 0 mL/min. The mobile phase is 85% CO 2 , 15% iPrOH (+ 0.3% iPrNH 2 ) held 7 min in isocratic mode.

[00332] Method 2: In addition to the general procedure: The analytical chiral separation in SFC-MS was carried out on a CHIRALPAK AD DAICEL column (10 μm, 4.6 x 250 mm) at 35°C with a flow rate of 3, 0 mL/min. The mobile phase is 75% CO 2 , 15% iPrOH (+ 0.3% iPrNH 2 ) held 7 min in isocratic mode.

[00333] Method 3: In addition to the general procedure: The analytical chiral separation in SFC-MS was carried out on a CHIRALPAK AD DAICEL column (10 μm, 4.6 x 250 mm) at 35°C with a flow rate of 3, 0 mL/min. The mobile phase is 80% CO2, 10% Methanol + 10% iPrOH (+ 0.3% iPrNH2) held for 7 min in isocratic mode. Table D: SFC Analytical Data - Rt means retention time (in minutes ), [M+H]+ means the protonated mass of the compound, method refers to the method used for SFC/MS analysis of enantiomerically pure compounds. The measurement was compared against the mixture. *A means the first eluting isomer. B means the second eluting isomer.

Nuclear Magnetic Resonance (NMR)

[00334] For a number of compounds, 1H NMR spectra were recorded on either a Bruker DPX-400 or a Bruker AV-500 spectrometer with standard pulse sequences, operating at 400 MHz and 500 MHz, respectively. Chemical shifts (δ) are reported in parts per million (ppm) in low field relative to tetramethylsilane (TMS), which was used as an internal standard. Co. At the. 6-b: 1H NMR (400 MHz, CDCl3) δ ppm 0.30 - 0.38 (m, 2H), 0.59 - 0.68 (m, 2H), 1.14 - 1.22 ( m, 1H), 1.72 (d, J=6.5 Hz, 3H), 3.02 - 3.14 (m, 2H), 5.84 (q, J=6.3 Hz, 1H), 6.67 - 6.73 (m, 1H), 6.80 - 6.89 (m, 2H), 7.30 (d, J=7.4Hz, 1H), 8 .11 (d, J=7.4 Hz, 1H) Co. At the. 7-b: 1H NMR (400 MHz, CDCl3) δ ppm 0.30 - 0.39 (m, 2H), 0.59 - 0.68 (m, 2H), 1.11 - 1.23 ( m, 1H), 1.70 (d, J=6.5 Hz, 3H), 3.01 - 3.14 (m, 2H), 5.83 (q, J=6.2 Hz, 1H), 6.35 - 6.45 (m, 3H), 7.13 (d, J=7.2 Hz, 1H), 8.08 (d, J=7.4 Hz, 1H ) Co. At the. 8-b: 1H NMR (400 MHz, CDCl3) δ ppm 0.30 - 0.38 (m, 2H), 0.58 - 0.68 (m, 2H), 1.11 - 1.22 ( m, 1H), 1.69 (d, J=6.2 Hz, 3H), 3.01 - 3.13 (m, 2H), 5.79 (q, J=6.2 Hz, 1H), 6.53 (ddd, J=9.2, 3.1, 3.1, 1.7 Hz, 1H), 6.72 (ddd, J=11.6, 6.5, 3 .1 Hz, 1 H), 6.95 - 7.04 (m, 1 H), 7.15 (d, J=7.4 Hz, 1 H), 8.07 (d, J=7.4 Hz, 1H) Co. At the. 15-b: 1H NMR (500 MHz, CDCl3) δ ppm 0.30 - 0.41 (m, 2H), 0.59 - 0.71 (m, 2H), 1.16 - 1.25 ( m, 1H), 1.70 (d, J=6.4 Hz, 3H), 3.05 - 3.16 (m, 2H), 5.80 (q, J=6.4 Hz, 1H), 6.62 - 6.70 (m, 2H), 7.45 (d, J=7.5 Hz, 1H), 8.16 (d, J=7.2 Hz, 1H ) Co. At the. 13-b: 1H NMR (500 MHz, CDCl3) δ ppm 0.27 - 0.39 (m, 2H), 0.58 - 0.67 (m, 2H), 1.12 - 1.21 ( m, 1H), 1.73 (d, J=6.4 Hz, 3H), 2.22 (s, 3H), 3.06 (qd, J=15.4, 6.6 Hz, 2H), 5.92 (q, J=6.4 Hz, 1 H), 6.71 (d, J=8.4 Hz, 1 H), 6.89 (dd, J=8.4, 1.4 Hz, 1 H), 7.18 (d, J=1.7 Hz, 1 H), 7.32 (d, J=7.2 Hz, 1 H), 8.07 (d, J =7.2 Hz, 1H) Co. At the. 14-b: 1H NMR (500 MHz, CDCl3) δ ppm 0.28 - 0.39 (m, 2H), 0.57 - 0.69 (m, 2H), 1.12 - 1.21 ( m, 1H), 1.70 (d, J=6.6 Hz, 3H), 2.31 (s, 3H), 3.01 - 3.12 (m, 2H), 5.79 (q, J=6.6 Hz, 1 H), 6.55 (dd, J=9.0, 4.3 Hz, 1 H), 6.69 (td, J=8.5, 3.0 Hz, 1H), 6.87 (dd, J=9.0, 2.9 Hz, 1H), 7.17 (d, J=7.5 Hz, 1H), 8.06 (d, J=7.2 Hz, 1H) Co. At the. 20-b: 1H NMR (500 MHz, CDCl3) δ ppm 1.22 (t, J=7.1 Hz, 3H), 1.72 (d, J=6.4 Hz, 3H), 3, 58 (q, J=7.1 Hz, 2 H), 5.03 - 5.10 (m, 2 H), 5.84 (q, J=6.5 Hz, 1 H), 6.67 - 6.74 (m, 1H), 6.81 - 6.88 (m, 2H), 7.34 (d, J=7.2 Hz, 1H), 8.40 (d, J=7 .5Hz, 1H) Co. At the. 22-b: 1H NMR (500 MHz, CDCl3) δ ppm 1.23 (t, J=6.9 Hz, 3H), 1.70 (d, J=6.4 Hz, 3H), 3, 58 (q, J=7.0 Hz, 2 H), 5.05 - 5.12 (m, 2 H), 5.81 (q, J=6.6 Hz, 1 H), 6.62 - 6.70 (m, 2H), 7.48 (d, J=7.5Hz, 1H), 8.45 (d, J=7.2Hz, 1H) Co. At the. 31-b 1H NMR (400 MHz, CDCl3) δ ppm 1.07 (t, J=7.40 Hz, 3H) 1.72 (d, J=6.24 Hz, 3H) 1.92 (sxt , J=7.63 Hz, 2H) 2.98 - 3.14 (m, 2H) 5.84 (q, J=6.47 Hz, 1H) 6.65 - 6.74 (m, 1H) 6.78 - 6.89 (m, 2H) 7.29 (d, J=7.40 Hz, 1H) 8.02 (d, J=7.40 Hz, 1H).

In vitro test of compounds of Formula (I-B)

[00335] The compounds of Formula (I-B) provided in the present invention are positive allosteric modulators of mGluR2. These compounds appear to potentiate glutamate responses by binding to an allosteric site other than the glutamate binding site. The mGluR2 response to a glutamate concentration is enhanced when compounds of Formula (I-B) are present. Compounds of Formula (I-B) are expected to have their effect substantially on mGluR2 by virtue of their ability to enhance receptor function. The effects of positive allosteric modulators tested on mGluR2 using the [35S]GTPYS binding assay method described below and which is suitable for the identification of such compounds, and more particularly of compounds according to Formula (I-B), are shown in Table E.

[35S]GTPYS binding assay

[00336] The [35S]GTPYS binding assay is a membrane-based functional assay used to study G protein-coupled receptor (GPCR) function whereby the incorporation of a non-hydrolyzable form of GTP, [35S] ]GTPYS (guanosine 5'-triphosphate, labeled with 35S gamma emitter), is measured. The α subunit of the G protein catalyzes the exchange of guanosine 5'-diphosphate (GDP) for guanosine triphosphate (GTP) and, upon activation of the GPCR by an agonist, [35S]GTPYS, becomes incorporated and cannot be cleaved to continue the exchange cycle (Harper (1998) Current Protocols in Pharmacology 2.6.1-10, John Wiley & Sons, Inc.). The amount of incorporation of radioactive [35S]GTPYS is a direct measure of G protein activity hence antagonist activity can be determined. The mGlu2 receptors are shown to preferentially couple to the Gαi protein, a preferred coupling for this method, and consequently it is widely used to study receptor activation of mGlu2 receptors in both recombinant cell lines and tissues. Here we describe the use of the [ 35 S]GTPYS binding assay using membranes from cells transfected with the human mGlu2 receptor and adapted from Schaffhauser et al. (Molecular Pharmacology, 2003, 4: 798-810) for detecting the positive allosteric modulation (PAM) properties of the compounds of this invention.

Membrane preparation

[00337] CHO cells were grown to preconfluence and stimulated with 5 mM butyrate for 24 h. Cells were then collected by scraping into PBS and the cell suspension was centrifuged (10 min at 4000 RPM in a benchtop centrifuge). The supernatant was discarded and the pellet carefully resuspended in 50 mM Tris-HCl, pH 7.4 by vortexing and pipetting up and down. The suspension was centrifuged at 16,000 RPM (Sorvall RC-5C plus SS-34 rotor) for 10 minutes and the supernatant discarded. The pellet was homogenized in 5 mM Tris-HCl, pH 7.4, using an ultra-turrax homogenizer and centrifuged again (18,000 RPM, 20 min, 4°C). The final pellet was resuspended in 50 mM Tris-HCl, pH 7.4 and stored at -80°C in appropriate aliquots before use. Protein concentration was determined by the Bradford method (Bio-Rad, USA) with bovine serum albumin as a standard.

[35S]GTPYS binding assay

[00338] Measurement of mGluR2 positive allosteric modulatory activity of test compounds was performed as follows. Test compounds and glutamate were diluted in assay buffer containing 10 mM HEPES acid, 10 mM HEPES salt, pH 7.4, 100 mM NaCl, 3 mM MgCh, and 10 µM GDP. Membranes containing human mGlu2 receptor were thawed on ice and diluted in assay buffer supplemented with 14 μg/mL saponin. Membranes were pre-incubated with compound alone or together with a predefined concentration (-EC20) of glutamate (PAM assay) for 30 min at 30°C. After addition of [35S]GTPyS (f.c. 0.1 nM), assay mixtures were shaken briefly and further incubated to allow incorporation of [35S]GTPYS after activation (30 minutes, 30°C). Final assay mixtures contained 7 µg of membrane protein in 10 mM HEPES acid, 10 mM HEPES salt, pH 7.4, 100 mM NaCl, 3 mM MgCl2, 10 µM GDP, and 2 µM saponin. µg/mL. The total reaction volume was 200 µL. Reactions were terminated by rapid filtration through Unifilter-96 GF/B plates (PerkinElmer, Massachusetts, USA) using a 96-well filtermate universal manifold. Filters were washed 6 times with ice-cold 10 mM NaH2PO4/10 mM Na2HPO4, pH 7.4. The filters were then air-dried, and 40 µL of liquid scintillation cocktail (Microscint-O) was added to each well. Membrane-bound radioactivity was counted on a PerkinElmer Microplate Scintillation and Luminescence Counter.

Data analysis

[00339] The concentration-response curves of representative compounds of the present invention - obtained in the presence of mGluR2 glutamate agonist EC20 to determine positive allosteric modulation (PAM) - were generated using the Lexis software interface ( developed at J&J). Data were calculated as % of control glutamate response, defined as the maximum response that is generated after addition of glutamate alone. Sigmoid concentration-response curves plotting these percentages versus log concentration of test compound were analyzed using nonlinear regression analysis. The concentration producing half-maximal effect is then calculated as the EC50.

[00340] The EC50 values below were calculated as the - log EC50, when the EC50 is expressed in M. Emax is defined as the relative maximal effect (i.e., the % maximal effect relative to the control glutamate response) .

[00341] Table E below shows pharmacological data obtained for compounds of Formula (IB) and current pharmacological data obtained for compounds of Formulas (I) and (IA). Table E: Pharmacological data for compounds according to the invention. nc means the pEC50 could not be calculated nt means not tested

[00342] The pEC50 values were not calculated in cases where the concentration response curve did not reach a plateau level.

[00343] All compounds were tested in the presence of the mGluR2 agonist glutamate at a predetermined EC20 concentration to determine positive allosteric modulation. The pEC50 values were calculated from a concentration-response experiment of at least 8 concentrations.

B) ANTICONVULSANT STUDIES WITH mGluR2 COMPOUNDS (ORTHOSTERIC AGONIST AND COMPOUNDS OF FORMULAS (I)/(I-A)/(I-B) General Preparation of test compounds and solutions

[00344] Test compounds were administered using an optimal ratio of fluid volume to body fluid. Test compounds were administered to mice in a volume of 0.01 mL/g of body weight (White, H.S., et al., General Principles: Experimental selection, quantification, and evaluation of antiepileptic drugs, in Antiepileptic Drugs, Fourth Edition , R.H. Levy, R.H. Mattson, and B.S. Meldrum, Editors. 1995, Raven Press, Ltd.: New York. pp. 99-110). For subcutaneous (s.c.) administration, test compounds were administered into a loose skin fold along the back of the animal except compound 6-b, which was administered orally (p.o). For each of the tests performed on test compounds (except compound 6-b), final compound concentrations were administered as an aqueous solution in 20% Hp-β-CD. For compound 6-b, a 40% Hp-β-CD stock solution was first prepared and used to formulate compound 6-b at the desired concentrations for testing via the oral route; final compound concentrations were administered as suspensions in 20% Hp-β-CD. A 20% Hp-β-CD solution was used for vehicle groups.

[00345] For LY-404039, final compound concentrations were administered as a saline s.c. solution.

[00346] For compound CAS 1092453-15-0, final compound concentrations were administered in 10% Hp-β-CD vehicle (+ NaCl) after dissolution.

[00347] The final concentrations of levetiracetam were administered in an aqueous solution of methylcellulose (MC) at 0.5 $ administered by intraperitoneal (i.p.) injection. Critical Reagents a) 0.5% Methylcellulose (MC) Vehicle Solutions 40% Hydroxypropyl-β-Cyclodextrin (Hp-β-CD) Stock Solution b) Miscellaneous Solutions

[00348] Tetracaine (0.5% w/v solution) was added dropwise from a plastic dropper bottle to the eyes of all animals that would subsequently receive electrical stimulation through corneal electrodes.

Animals and Animal Husbandry

[00349] CF #1 adult male albino mice (26-35 g) were obtained from Charles River, Portage, Michigan. Animals were maintained on an adequate diet (Prolab RMH 3000) and had free access to food and water, except for the short time they were removed from their cage for testing. Animals newly received at the laboratory were given sufficient time to correct possible food and water restriction incurred during transit before being employed for testing. All mice were housed in plastic cages in specially constructed chambers with controlled humidity, air exchange and controlled lighting (12 hours light - 12 hours no light). Animals were housed, fed, and handled in a manner consistent with recommendations in the National Council Publication, "Guide for the Care and Use of Laboratory Animals".

Minimum Motor Commitment (MMI)

[00350] Acute MMI was assessed by a combination of direct observations of the animal for overt symptoms of the animal's neurological or muscular function. In mice, the rotarod procedure was used to reveal minimal muscular or neurological impairment. When a mouse is placed on a rod that rotates at a speed of 6 rpm, the animal can maintain its balance for long periods of time. The animal was considered toxic if it fell from this rotating rod three times during a period of 1 min.

Determination of Median Effective and Toxic Doses (ED50 and TD50)

[00351] In determining an ED50 or TD50 for each test compound, the first dose administered is usually the same dose as that used in a successful TPE determination. If the initial dose employed was effective or toxic in more than 50% of the animals, the next dose would be half that of the initial dose; if the initial dose was effective or toxic in less than 50% of the animals, the next dose would be twice that of the initial dose. The third and fourth doses were selected to produce an evenly spaced dose-response line. There must be a minimum of 4 points including or being between 0 to 100%.

TPE determination

[00352] Groups of typically four animals each were given test compounds and each group was tested at one of five time points: 0.25, 0.5, 1, 2, or 4 h post-treatment (White et al. 1995). TPE was determined using the 6 Hz (32 mA) assay. The time (0.25, 0.5, 1, 2, or 4 h post-treatment) at which maximum protection was observed was considered the Time of Peak Effect (TPE).

[00353] At the TPE determined for this study, or previously determined, the compounds were tested in the 6 Hz assay (32 and/or 44 mA), over several doses and comprising doses that caused little or no protection to complete protection.

[00354] An ED50 and 95% confidence interval (CI) were calculated using Probit analysis in a computer program provided in the laboratory (Finney "Probit Analysis" 34th Ed 1971, London: Cambridge University Press).

Serum Collection for pK/pD Analysis

[00355] In several tests, animals were sacrificed after testing, and blood from stem and/or brain tissue (whole brains) was collected for quantification of drug levels. Immediately after testing, the animals were decapitated and trunk blood was collected in a BD Vacutainer® tube containing K2EDTA and chilled on ice until centrifugation. After centrifugation (13000 - 18000 rpm, 5-7 min), the plasma was removed and transferred to a labeled microcentrifuge tube and stored at -80°C. For brain tissue collection, brains were removed immediately after decapitation and immediately frozen. The frozen sample was placed in a labeled centrifuge tube and stored at -80°C.

6 HZ Psychomotor Seizures Test in Mice

[00356] The 6 Hz Seizures test is used as a model of drug-resistant limbic seizures. The 6 Hz seizure exhibits a resistance to phenytoin, carbamazepine, lamotrigine, and topiramate (Barton et al. "Pharmacological characterization of the 6 Hz psychomotor seizure model of partial epilepsy" Epilepsy Research 2001, Vol. 47, pp. 217-222).

Method for Testing Psychomotor Seizures 6 Hz

[00357] Focal seizures were induced in mice by corneal stimulation (6 Hz, 0.2 msec rectangular pulse, 3 sec duration; Barton et al., 2001). Mice were tested at either 32 mA or 44 mA. Before stimulation, 0.5% tetracaine drops were applied to each eye. Seizures arising from corneal stimulation in this trial are characterized by a minimal clonic phase followed by stereotyped automatism behaviors including stunting, forelimb clonus, vibrissae twitching, and Straub's tail. Animals not exhibiting these behaviors were considered protected.

EXAMPLE 1 - STUDIES WITH COMPOUNDS 1 AND 2 1.1. Combination study with Co. At the. 1, Co. At the. 2 and Levetiracetam

[00358] First, each compound was individually tested at a dose that exhibited minimal activity in the 44 mA 6 Hz TPE test of each compound. When mGluR2 PAM compounds and levetiracetam were administered in combination (same dose and time point as the individual tests) almost complete protection was observed in the 44 mA 6 Hz test (Table 2). In addition to recording efficacy and toxicity data for these compounds alone or in combination, both plasma and brain samples were collected from each of the groups for pharmacokinetic analysis. No pharmacokinetic interactions were observed based on compound levels in plasma and brain samples (data not shown). In summary, compounds 1 and 2 exhibited positive pharmacodynamic interaction with levetiracetam in the 6 Hz model that does not appear to be due to pharmacokinetic interaction, and without increased motor impairment (Tables 2, 2a, 2b). The effect of 1 dose of Compound 2 was also tested on the dose response of LEV. As shown in Table 3 there was a -200 fold shift in the ED50 of LEV compared to when LEV was tested alone. LEV appeared to increase the potency of Co No. 2 slightly (Table 3).

1.2. Isobolographic analysis of interactions between Co. At the. 1 and Leveti-racetam in the 6 Hz seizure model

[00359] Isobolographic studies were conducted for the combined administration of Co. At the. 1 with LEV in the 6 Hz test (44 mA). Studies were conducted according to previously described methods (Madsen et al. 2011). Initial ED50 values were determined for Co. At the. 1 and LEV and used to calculate theoretical ED50 values (+ standard error of mean, SEM) for three fixed dose ratio combinations (LEV: Co. No. 1): 1:3, 1:1, and 3: 1. The doses used were proportionate to the calculated ED50 values. For example, the dose ratio used for the 1:1 paradigm was based on 0.5 x ED50 for LEV and 0.5 x ED50 for Co. At the. 1. Similarly, the 1:3 paradigm used 0.25 x ED50 for LEV and 0.75 x ED50 for Co. At the. 1. The 3:1 dose ratio used 0.75 x ED50 for LEV and 0.25 x ED50 for Co. At the. 1. Experimental treatment doses (see Table 4) were based on theoretical values and adjusted according to observed effects. The determined experimental ED50 values (+ SEM) for each fixed dose ratio combination were compared with the theoretical values (t-test) for statistical purposes. The dose ratio was determined to be supra-additive (synergistic) if the experimentally determined ED50 value was significantly lower than the theoretical ED50. Subsequently, experimental combined doses were determined for the same paradigms in the 6 Hz seizure test (Table 4 below). The isobolographic study with compound 1 and levetiracetam in the 6 Hz model demonstrates a significant synergistic pharmacodynamic interaction at all dose ratios evaluated and closely matches the plasma levels of compound 1. Furthermore, no motor impairment was observed at any time. one of the evaluated dose ratios suggesting that the synergistic pharmacodynamic interaction does not produce increased motor toxicity.

1.3. Mice Corneal Burnishing Model and studies with compound 1

[00360] The mice were electrically abraded with a stimulus of 60 Hz, 3 mA, 3 seconds, twice daily using corneal electrodes up to a criterion of 5 consecutive Step 5 seizures as defined by Racine (Racine "Modification of seizure activity by electrical stimulation" II. motor seizure" Electroenceph Clin Neurophysiol 1972, 32, pp. 281-294). each animal the electrical stimulus indicated above. After stimulation, the animals were observed for the presence or absence of seizure activity scored on the Rancine scale (0 - 5) with 5 representing the rising and falling of higher step. dose of LEV and two doses of Co. No. 1 were tested individually and in combination against corneal-flaring seizures.The combination of compound 1 with levetiracetam in this model suggests a positive pharmacodynamic interaction (Table 5 below).

[00361] A summary of data for the compounds tested alone is presented in Table 1 and additional results from studies performed according to example 1 are listed in Tables 2-5 below.Table 1: Summary of acute anticonvulsant data in the model of 6 Hz at 32 mA and 44 mA for mGluR2 PAM compounds 1, 2, 11, 2-a, 25-a, 6-b and LY-404039 after sc administration (except compound 6-b, which was tested po) . TPE means time of effect at peak, CI means confidence interval, sc means subcutaneous, po means orally, nt means not tested. The TPE was determined in the 6 Hz 32 mA test. Effects are generally seen at doses that do not produce impairment in the rotarod test. For compounds 11 and 2-a, individual values from repeated experiments are provided. For compound 25a, both the 0.25 and 1 h time points were used for 6 Hz (44 mA) studies. Table 1a: Summary of TPE determination at 32 mA 6 Hz for Co. At the. 1 (number of mice protected at 6 Hz or toxic in rotarod/number tested) Table 1b: Dose-response studies for Co. At the. 1 The TPE for Co. At the. 1 was previously determined to be 0.5 h (results shown above in table 1a). Several doses of Co. At the. 1 were administered to this TPE and tested in the 6 Hz assay, using stimulus intensities at 32 and 44 mA. Table 2: Summary of Co interaction. At the. 1 and Co. At the. 2 with Leve-tiracetam (LEV) in the mouse model of seizures 6 Hz, 44 mA. Results are listed as number of mice exhibiting total protection/total number of mice tested in each dose group (at the specified test compound or combination dose levels). Table 2a: Plasma and brain levels Co. At the. 1. in combination study with Levetiracetam (LEV). BQL means below the quantifiable limit. The mean plasma level shown in parentheses ( ) is calculated with a statistic* outlier removed. Table 2b: Plasma and brain levels Co. At the. 2. in combination study with Levetiracetam (LEV). AQL stands for above quantifiable threshold. NA - sample not available for analysis Table 3: Seizure Model ED50 determinations (44 mA) 6 Hz for No. Co. 2 and Levetiracetam (LEV) alone and in combination. LEV at a dose of 10 mg/kg increased the potency of No. Co. 2 (~5 fold deviation at ED50). Co. At the. 2 at a dose of 3 mg/kg increased both the efficacy (up to 100% protection) and the potency of LEV (~200-fold deviation in ED50). Figure 1 shows the dose response for 44 mA 6 Hz ED50 determinations for Co. At the. 2 and LEV alone and in combination. Table 4: Results from Co. At the. 1 and Levetiracetam in an Isobolographic Study. The isobolographic analysis (Figure 2) demonstrates that the combination of Co. At the. 1 and Levetiracetam results in a significantly positive synergistic effect. Table 5: Co combination study results. At the. 1 and Le-vetiracetam in the corneal kindling model in mice.

EXAMPLE 2 - STUDIES WITH COMPOUNDS 25-a AND 2-a 2.1. Combination study with Co. At the. 25-a and Levetiracetam

[00362] Independent dose-response studies were performed on the 6 Hz 44mA test for both compounds to determine ED50 values at 1 h ip TPE for levetiracetam and 1 h sc for Co. At the. 25-a. The ED50 value for Co. At the. 25-a was 25.9 mg/kg and for levetiracetam the value was estimated to be approximately 345 mg/kg. The dose response for levetiracetam was repeated with co-administration of 10 mg/kg Co. At the. 25-a (a dose of Co. No. 25-a alone did not protect in the 44 mA 6 Hz model). Co-administration of 10 mg/kg of Co. At the. 25-a produced an ED50 on dose response of levetiracetam of 4.9 mg/kg (~70-fold lower compared to levetiracetam alone) and importantly gave complete protection in the 44 mA 6 Hz seizure model. These results are suggestive of a positive pharmacodynamic interaction in the 6 Hz seizure model between Co. At the. 25-ae levetiracetam. Table 6: Determination of Peak Effect Moment for Co. At the. 25-a in the 6 Hz Test (32 mA). Two doses were used in this study, 10 and 20 mg/kg, over various time points (0.25 - 4 h). The compound showed the greatest degree of protection in the 6 Hz test between 0.25 and 1 h, which was most evident at 20 mg/kg. Plasma levels of the compound generally corresponded to protection from behavioral seizures. A TPE of 0.25 was used for 6 Hz (32 mA) studies while 0.25 and 1 h time points were used for 6 Hz (44 mA) studies. sc means subcutaneous The mean plasma level shown in parentheses ( ) is calculated with a statistical outlier removed. Table 7: Dose Response Studies for Co. At the. 25-a in the 6 Hz Assay (32 mAa and 44 mAb) aThe time of effect at the peak in the 32 mA 6 Hz assay for Co. At the. 25-a was determined to be 0.25 h (see Table 1). bThe time of effect at the peak in the 44 mA 6 Hz assay for Co. At the. 25-a was similar for 0.25 h and 1 h; results for 1 h confirmed the ED50 (95% CI) 25.9 (15.5 - 33.7) (see Tables 1 and 6).

[00363] The mean plasma level shown in parentheses ( ) is calculated with a statistical outlier removed.Table 8: Combination Studies for Co. At the. 25-a with Leveti-racetam (LEV) in the 6 Hz Assay (44 mA). Co. At the. 25-a (sc) 10 mg/kg tested in combination with LEV (ip) - Co. At the. 25- to 10 mg/kg, not active when given alone.

2.2. Combination study with Co. At the. 2-a and Levetiracetam

[00364] Dose-response studies were performed in the 32 mA and 44 mA 6 Hz tests (table 9 below) and in the combination test with levetiracetam (effect of Co. No. 2-a on the dose response of LEV in the tables 10a and effect of LEV on the dose response of Co. No. 2-a in Table 10b below) in the same manner as described for the Co studies. At the. 25-a and levetiracetam alone. Table 9: Dose Response Studies for Co. At the. 2-a in the 6 Hz Run (32 mA and 44 mA; 0.5 h TPE). A peak time of effect of 0.5 h was determined in the 32 mA 6 Hz (sc) test and used for the 6 Hz studies (32 mA and 44 mA). a40 mg/kg - 6 of 16 total (32 mA and 44 mA combined) with impairment. Dose selected for combination studies with LEV at 6 Hz (44 mA): Co. At the. 2- to 10 mg/kg. Table 10a: Combination Studies for Co. At the. 2-a with Leveti-racetam (LEV) in the 6 Hz Test (44 mA). Combination of 10 mg/kg of Co. At the. 2-a with varying doses of levetiracetam. steel. At the. 2-a (sc) 10 mg/kg tested in combination with LEV (ip); Co. At the. 2-to 10 mg/kg, not active when given alone.

[00365] Additional LEV (low dose) control groups were tested at 25 and 6.25 mg/kg (1/8 and 0/6 protected, respectively).

[00366] Vehicle-treated mice (methylcellulose 0.5% ip (1 h) / HPBCD 20% sc (0.5 h)) showed no protection (0/8 protected). Table 10b: Combination Studies for Co. At the. 2-a with Leveti-racetam (LEV) in the 6 Hz Test (44 mA). Combination of 350 mg/kg of levetiracetam with varying doses of Co. At the. 2-a. ED50 preview from Co. At the. 2-a (95% CI): 21.0 mg/kg (17.9 to 27.4) ED50 prior to combination of LEV and Co. At the. 2-a (95% CI): 1.5 mg/kg (0.1 - 2.7) ~14-fold deviation in previously determined LEV aED50 potency (shown separately) at 6 Hz (44 mA): 345 mg/kg. bCo. At the. 2-a (sc) 10 mg/kg tested in combination with LEV (ip); Co. At the. 2-to 10 mg/kg, not active when given alone.

[00367] Additional LEV (low dose) control groups were tested at 25 and 6.25 mg/kg (1/8 and 0/6 protected, respectively). Mice treated with vehicle (0.5% methylcellulose i.p. (1 h) / 20% s.c. HPBCD (0.5 h)) showed no protection (0/8 protected).

[00368] At a dose of 10 mg/kg s.c., Co. At the. 2-a increases LEV power, leading to an approximately 35-fold shift in ED50. This suggests a positive pharmacodynamic relationship (Table 10a). At a dose of 350 mg/kg i.p., LEV increases Co potency. At the. 2-a, leading to an approximately 14-fold shift in the ED50. This suggests a positive pharmacodynamic relationship (Table 10b).

EXAMPLE 3 - STUDIES WITH COMPOUND 6-b 3.1. Combination study with Co. At the. 6-b and Levetiracetam

[00369] Independent dose-response studies were performed on the 6 Hz 44mA test for both compounds to determine ED50 values at 1 h i.p. for levetiracetam and 0.5 h i.p. to Co. At the. 6-b. The ED50 value for Co. At the. 6-b was 16.1 mg/kg and for levetiracetam the value was estimated to be approximately 345 mg/kg. The dose response for levetiracetam was repeated with co-administration of 10 mg/kg Co. At the. 6-b (a dose of Co. No. 6-b which alone did not protect in the 44 mA 6 Hz model). Co-administration of 10 mg/kg of Co. At the. 6-b produced an ED50 on dose response of levetiracetam of 2.4 mg/kg (~100-fold lower compared to levetiracetam alone) and importantly gave complete protection in the 44 mA 6 Hz seizure model. These results are suggestive of a positive pharmacodynamic interaction in the 6 Hz seizure model between Co. At the. 6-b and levetiracetam.

[00370] The results of studies performed with compound 6-b are listed in Tables 11-13 below. Table 11: Determination of Peak Effect Timing for Co. TPE determined to be 0.5 h. Table 12: Dose Response Study for Co. At the. 6-b in the 6 Hz Run (32 mA and 44 mA; 0.5 h TPE). Table 13: Combination Studies for Co. At the. 6-b with LEV in the 6 Hz Test (44 mA). Co. At the. 6-b (po) 10 mg/kg tested in combination with LEV (ip) Co. At the. 6-b 10 mg/kg, not active when given alone.

EXAMPLE 4 - STUDIES WITH COMPOUND LY-404039 3.1. Combination Study with LY-404039 and Levetiracetam

[00371] LY-404039 was tested alone and in combination with mild tiracetam according to the procedures already described above. Results of studies performed with LY-404039 are listed in Tables 14-15. Table 14: Dose Response Studies for LY-404039 in the 6 Hz Assay (32 mA and 44 mA). A peak time of effect of 0.5 h was determined in the 32 mA 6 Hz (sc) test and used for the 6 Hz studies (32 mA and 44 mA). a40 mg/kg - 3 of 16 total (32 mA and 44 mA combined) with impairment. Note: No activity observed after administration of vehicle at 32 or 44 mA.

[00372] Selected dose for combination studies with LEV at 6 Hz (44 mA): LY-404039 5 mg/kg. Table 15: Combination Studies for LY-404039 with Levetira-cetam (LEV) in the 6 Hz Assay (44 mA). aLEV alone shown previously, confirmation doses performed in combination with Co. At the. 2-a (see previous table above). bLY-404039 (sc) 5 mg/kg tested in combination with LEV (ip); LY-404039 5 mg/kg is not active when given alone.

[00373] Additional LEV (low dose) control groups were tested at 25 and 6.25 mg/kg (1/8 and 0/6 protected, respectively).

[00374] Vehicle-treated mice (10% sterile water - BR112016016390-7 Janssen Pharmaceutica NV P221633 protection or impairment in rotarod.

[00375] At a dose of 5 mg/kg, LY-404039 increases LEV potency, leading to an approximately 27-fold shift in ED50. This suggests a positive pharmacodynamic relationship. EXAMPLE 5 - REFERENCE STUDIES WITH COMPOUND CAS 1092453-15-0 5.1. Combination Study with 2,3-dihydro-7-methyl-5-[3-(1-piperazinylmethyl)-1,2,4-oxadiazol-5-yl]-2-[[4-(trifluoromethoxy)phenyl]methyl ]-1H-isoindol-1-one [CAS 1092453-15-0] (described in WO 2008150233, WO 2011084098) and Levetiracetam

[00376] CAS 1092453-15-0 was tested alone and in combination with levetiracetam according to the procedures already described above. The results for example 5 are listed in tables 1617. Table 16: Dose Response Studies for CAS 1092453-15-0 in the 6 Hz Assay (32 mA).

[00377] Low activity was observed at the doses and time points tested. Greater activity at 0.25-1 h at the doses tested. Combination studies were performed using 20 mg/kg, sc, 1 h TPE in the 6 Hz Assay (44 mA). Table 17: Combination Studies for CAS 1092453-15-0 with Levetiracetam (LEV) in the 6 Hz Assay (44 mA bad). aAdditional LEV (low dose) control groups were tested at 25 and 6.25 mg/kg (1/8 and 0/6 protected, respectively). b[CAS 1092453-15-0] 20 mg/kg (sc; 1 h EPT) tested in combination with LEV (ip; 1 h EPT); [CAS 1092453-15-0] 20 mg/kg exhibited low activity when administered alone (6 Hz, 32 mA), and was not tested at 6 Hz (44 mA). This compound exhibited an in vitro EC50 = 562 nM (Emax = 197%) when tested in the GTPYS assay described above and no occupancy was observed in ex vivo experiments in mice. Note: Vehicle-treated animals (10% HPβCD- NaCl, sc, 1 h and 0.5% MC, ip, 1 h) showed no rotarod protection or impairment, N = 8.

[00378] The current dataset indicates that PAM molecules or mGlu2 agonists have anticonvulsant activity in the 6 Hz animal model. mGlu2 PAMs tested with potencies of EC50 <150 nM (as determined in the [35S]GTPγS assay), PK parameters, and brain penetration, showed activities in both the 6 Hz 32 and 44 mA paradigms. Furthermore, all molecules tested showed synergistic effects with LEV. In contrast, the CAS 1092453-15-0 molecule, which was only weakly active (EC50 562 nM) in vitro, showed no activity in any of the 6 Hz assays, and also did not exhibit synergy with LEV.

[00379] Importantly, the data indicate that, under conditions of comparable PK characteristics and adequate brain penetration, the most potent mGlu2 PAMs, based on in vitro EC50 values, also appeared the most potent in vivo, suggesting that in vitro potency and in vivo it may be on. Furthermore, synergistic effects with LEV were consistently seen with MAP doses of mGlu2 similar to the ED50 obtained in the 32 mA model or at least 2-fold lower than the ED50 determined in the 44 mA paradigm (i.e., an inactive dose in the 32 mA test). 44 mA when the molecules were tested alone).

[00380] Also for LY-404039, the mGlu2/3 agonist, activity was seen in both 6 Hz tests and synergy was seen at a dose 3 times lower than the ED50 determined in the 44 mA model, which was inactive when tested alone.

[00381] Based on preclinical data available in the 44 mA 6 Hz model it appears that the combination of a potent SV2A ligand and a potent mGlu2 MAP leads to a decrease in the median effective dose or ED50 of the SV2A ligand, as as LEV, between 35 and 100 times.

[00382] Thus, while not wishing to be bound by theory, it is suggested that positive allosteric modulator compounds of the metabotropic glutamatergic receptor subtype 2 (mGluR2 PAM), in particular mGluR2 PAM compounds having an EC50 potency of <150 nM (as determined in the [35S]GTPγS assay), where EC50 is the concentration producing half-maximal effect on a dose-response curve obtained in the presence of glutamate EC20, and appropriate PK parameters and brain penetration, result in a synergistic combination with an SV2A binder, in particular levetiracetam, at non-effective doses of one or both of compound (a) and compound (b) of the combination of the invention.

[00383] Thus, in a further embodiment, the metabotropic glutamatergic receptor subtype 2 (mGluR2 PAM) positive allosteric modulator compound of the combination of the invention as defined herein is selected from a mGluR2 PAM compound having an EC50 potency of <150 nM (as determined in the [35S]GTPγS assay), where EC50 is the concentration producing half-maximal effect on a dose-response curve obtained in the presence of glutamate EC20.

PROPHETIC EXAMPLES A) DOMINANT-RECESSIVE RATIOS (DSR) IN IN VIVO TEST OF RATS

[00384] The DSR trial is divided into two models: Reduction of the Dominant Behavior Model (RDBM) of mania and Reduction of the Submissive Model of Behavior (RSBM) of depression. The RDBM, in which dominant animals are treated with the test compound, is predictive of the ability of the test compound to treat mania. The RSBM, in which compliant animals are treated with the test compound, is predictive of the ability of the test compound to treat depression.

[00385] Male Sprague Dawley rats (140 to 160 g) from Charles River Laboratories Wilmington, MA are used in this assay. Rat shipments are received every 2 weeks. Each shipment will undergo a five-day quarantine, one-week acclimatization period, and one-week selection process, followed by five weeks of drug or vehicle treatment for those selected pairs.

[00386] The rats will be housed four per cage. Access to food will be restricted to one hour a day after testing on Monday through Thursday. After testing on Friday, the rats will have free access to food until they are fasted again on Sunday. Rats will never be deprived of water. The water deprivation periods used will have little effect on weight gain as the average rat weight will be around 300 g at the end of the study. At the conclusion of the experiment, mice will be sacrificed by decapitation, stem blood and brains will be collected for in vitro experiments and drug concentration measurements.

[00387] The basic test device consisted of two connected chambers with a tunnel just wide enough to allow one mouse to pass through at a time. On the floor at the midpoint of the tunnel will be a container of sweetened milk. This basic device will be replicated such that a total of four pairs of mice can be videotaped simultaneously. The camera can distinguish mice marked with different colors. Thus, the rats' heads will be colored for the purpose of video recording, red in one cage and yellow in the other cage. Only one animal at a time can access the feeder, but both animals can drink milk during the daily five-minute session. During daily five-minute sessions, the time spent in the feeder zone by each rat will be recorded by video recording software and saved to a text file.

[00388] The test will begin with a random assignment of mice in pairs. Each member of a pair will be placed in an opposite chamber of the test device. The time spent in the feeder zone by each animal will be recorded. During the first week (five days) of the test, the animals get used to the new environment. Dominance will be assigned to the animal with the highest score during the second week of testing if three criteria are met. First, there must be a significant difference (two-tailed t-test, P<0.05) between the mean daily drinking scores of both animals. Second, the dominant animal's score must be at least 25% higher than the submissive animal's score. Finally, there can be no "inversions" during the pair selection week where the putative submissive rat scores better than its dominant partner on isolated occasions. Ideally there will be minimal inversions during the acclimatization week as well. Only pairs of animals that meet these criteria will continue in the study.

[00389] Significant differences between the time spent on the feeder by dominant and submissive rats will be determined by ANOVA using GraphPad Prism software (GraphPad Software, Inc. San Diego, CA) followed by a two-tailed t-test (P<0 .05). Comparisons will be made between treatment groups using values of normalized dominance levels in the paired animals. The level of dominance is a value that measures the social relationship between paired individuals. Dominance Level (DL) = FTD-FTS where FTD is time at feeder for dominant rats and FTS is time at feed for submissive rats. Normalization will be conducted according to the formula: Dominance Level (week n in %) = (Dominance Level (week n)) / (Dominance Level (week 2))

[00390] The statistical significance of the difference in the level of dominance between the control group (the pairs of mice where both the dominant and submissive animals will be treated with vehicle) and the treatment group (the submissive mice will be treated with drug and vehicle-dominant rats) will be determined by ANOVA followed by a t-test. The activity onset time value at 50% response (AOT-50) and the minimum and maximum drug response will be calculated based on the reduction of the dominance level value using non-linear regression analysis (GraphPad Software, Inc. , San Diego, CA). The normalized LD values will be used for this calculation, where the LD values for the treatment weeks will be normalized as a percentage of the second week (pre-treatment) value of that pair according to the above formula. In these scenarios, the minimum response (DL) determines the positive activity of the drug, corresponding to efficacy, since DL values will be reduced if the response to a drug is positive. In case of negative response to a drug (worsening of symptoms), DL values will be increased. If the drug has no such activity, the maximum response will not exceed 100%. Any maximum LD value significantly higher than the control value (about 100%) indicates negative drug activity.

[00391] Levetiracetam and compound PAM/mGluR2 agonist (eg compounds 2, 2-a, 25-a, 6-b or LY-404039) will be evaluated in the RDBM of rats according to the procedure described in more detail below.

[00392] Groups of dominant rats will be treated p.o. QD with levetiracetam 10 mg/kg and PAM/mGluR2 agonist compound at various concentrations from approximately 0.05 mg/kg (n > 3), to 0.5 mg/kg (n > 3), to 2.5 mg/kg kg (n > 3), at 5.0 mg/kg (n > 3) and at 50.0 mg/kg (n > 3). A vehicle control group of dominant rats will be treated with 0.5% methylcellulose (n > 3) and a second control group of dominant rats will be treated i.p. QD with sodium valproate 30 mg/kg (n > 6 of 2 studies of n > 3 each).

[00393] All treatments will be given approximately 1 hour before the test. All treatments will start on the Saturday after the second test week (selection week). Levetiracetam and PAM/mGluR2 agonist compound will be administered orally (p.o.).

[00394] When dominant animals are treated with levetiracetam 10 mg/kg and PAM/mGluR2 agonist compound, the difference between dominant and submissive mice will be lost after the first or second week of treatment depending on the dosage. Similarly, when dominant animals are treated with sodium valproate, the difference between dominant and submissive rats will also be lost after the first week of treatment. It can be seen that the permissiveness of dominant rats treated with levetiracetam and PAM compound/mGluR2 agonist or sodium valproate increases. Thus, treated dominant rats will allow their submissive partners to increase their time at the feeder.

[00395] To compare different drug and dose effects, the data will be normalized in relation to the baseline values of the control week. The strongest effect of the combination of levetiracetam and compound PAM/mGluR2 agonist will be seen where there is a significant difference in dominance level (DL) values between vehicle and combination treated rats starting in the second week and continuing for the duration of treatment of 5 weeks. In comparison, animals (30 mg/kg) that will be treated with sodium valproate will consistently show a decreased dominance level after the second week of treatment with the effect increasing in the following weeks.

[00396] To estimate the time to onset of activity (AOT), the average daily values for the time at the feeder of pairs of dominant and submissive animals will be graphically represented and the significant differences between these two groups will be calculated using the t test of two tails.

[00397] To compare the time to onset of activity (AOT) between different treatments, the time to onset of activity will be estimated from the non-linear regression fit. The nonlinear regression model will adjust for each drug, combination, and dose-normalized daily LD value.

[00398] The effects of levetiracetam and compound PAM/mGluR2 agonist are expected to be dose-dependent.

[00399] In this trial the combination of levetiracetam and PAM compounds/mGlu2 agonists is expected to reduce dominant behavior indicating that the combination is active as an antimanic.

B) ORAL TABLETS

[00400] As a specific example of an oral composition, 100 mg of a PAM/mGluR2 agonist compound is formulated with sufficiently finally divided lactose to provide a total amount of 580 to 590 mg to fill a size O hard gel capsule.

[00401] Although the foregoing specification teaches the principles of the present invention, with examples provided for the purpose of illustration, it will be understood that practice of the invention encompasses all usual variations, adaptations and/or modifications as are within the scope of the following claims and their equivalents.

Claims (21)

1. A combination comprising (a) a synaptic vesicle protein 2A ("SV2A") ligand selected from the group consisting of levetiracetam, brivaracetam and seletracetam; and (b) an orthosteric agonist of the metabotropic glutamatergic receptor compound subtype 2 selected from i) LY-404039 or a salt thereof; and ii) LY-2140023 or a salt or a monohydrate thereof. 2. Combination according to claim 1, characterized in that the SV2A ligand is levetiracetam or brivaracetam. 3. Combination according to claim 1 or 2, characterized in that the metabotropic glutamatergic receptor subtype 2 orthosteric agonist is LY-404039 4. Combination according to claim 2, characterized in that the metabotropic glutamatergic receptor subtype 2 orthosteric agonist is the hydrochloride salt of LY-404039. 5. Combination according to claim 1 or 2, characterized in that the metabotropic glutamatergic receptor subtype 2 orthosteric agonist is LY-2140023 6. Combination according to claim 2, characterized in that it is for use in the treatment of epilepsy. 7. Combination according to claim 2, characterized in that the metabotropic glutamatergic receptor subtype 2 orthosteric agonist is LY-2140023 monohydrate. 8. Pharmaceutical composition, characterized in that it comprises a combination as defined in any one of claims 1 to 7, and a pharmaceutically acceptable vehicle. 9. Composition according to claim 8, characterized in that it is for use in the treatment of bipolar disorders. 10. Process for preparing the pharmaceutical composition as defined in claim 8, characterized in that the combination as defined in any one of claims 1 to 7 is intimately mixed with a pharmaceutically acceptable vehicle. 11. Product, characterized in that it comprises the combination of the SV2A ligand and the metabotropic glutamatergic receptor subtype 2 orthosteric agonist as defined in any one of claims 1 to 7, which are present in separate dosage forms or formulations for the treatment or prevention of epilepsy and related disorders; neuropathic pain; migraine or resistant headache; bipolar and related disorders. 12. A combination according to any one of claims 1 to 7, or a pharmaceutical composition according to claim 8, characterized in that it is for use as a medicine. 13. Combination according to any one of claims 1 to 7 or pharmaceutical composition according to claim 8, characterized in that it is for use in the treatment or prevention of epilepsy and related disorders; neuropathic pain; migraine or resistant headache; and bipolar and related disorders. 14. Combination or pharmaceutical composition according to claim 13, characterized in that it is for use in the treatment or prevention of epilepsy and related disorders. 15. Combination or pharmaceutical composition according to claim 14, characterized in that it is for use in the treatment or prevention of epilepsy. 16. Combination or pharmaceutical composition according to claim 15, characterized in that the epilepsy is treatment-resistant epilepsy. 17. Combination or pharmaceutical composition, according to claim 15, characterized in that the epilepsy is epilepsy with partial seizures with or without generalization. 18. Combination or pharmaceutical composition, according to claim 15, characterized in that the epilepsy is epilepsy with generalized seizures. 19. Combination according to claim 15, characterized in that the epilepsy is epilepsy with primary generalized tonic-clonic seizures. 20. Combination according to any one of claims 1 to 7, or a pharmaceutical composition according to claim 8, characterized in that it is for use in preventing epileptogenesis. 21. Pharmaceutical product according to claim 11, characterized in that it comprises instructions.