Nanoparticles for use for treating neuronal disorder

Abstract

To provide a nanoparticle or nanoparticles' aggregate for use in the medical field, in particular for the treatment of neurological disorders, for use in prevention or treatment of a neurological disease or at least one symptom thereof in a subject without exposure of the nanoparticle or nanoparticles' aggregate to an electric field, and preferably without exposure thereof to any other external activation source.SOLUTION: A material of a nanoparticle or nanoparticles' aggregate is selected from a conductor material, a semiconductor material, an insulator material having a dielectric constant εijk equal to or above 200, and an insulator material having a dielectric constant εijk equal to or below 100. Furthermore, a composition comprising the nanoparticle and / or nanoparticles' aggregate and a pharmaceutically acceptable carrier is provided, the composition being for use in prevention or treatment of a neurological disease or at least one symptom thereof in a subject without exposure of the nanoparticle and / or nanoparticles' aggregate to an electric field, and preferably without exposure thereof to any other external activation source.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to the medical field, in particular to the treatment of neurological disorders. More specifically, the present invention relates to nanoparticles or nanoparticle aggregates for use in the prevention or treatment of neurological disorders or at least one symptom thereof in a subject, without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source, wherein the material of the nanoparticles or nanoparticle aggregates is a conductive material, a semiconducting material, a material having a dielectric constant ε of 200 or more, ijk Insulating materials with a dielectric constant ε of 100 or less ijk Furthermore, the present invention relates to compositions and kits comprising such nanoparticles and / or nanoparticle aggregates, and their uses, without exposure to an electric field, and preferably without exposure to any other external activation source, such as a light source, a magnetic field, or an ultrasound source.

[0002] background Neurological disorders are a major health concern (Neurological disorders public health challenges. WHO, 2006). Impairments in neural network function can have different origins. Parkinson's disease is a movement disorder caused by the death of dopamine neurons in the substantia nigra, located in the midbrain. Stroke corresponds to the interruption of the brain's blood supply. Neurons in the affected area die due to oxygen deprivation, and the parts of the body controlled by these cells are unable to function. Huntington's disease is a genetic disorder. Epilepsy is a disorder caused by the abnormal excitation of many groups of neurons in various brain regions. Alzheimer's disease is a neurodegenerative disorder characterized by the death of neurons in the hippocampus, cerebral cortex, and other brain regions. The causes of autism spectrum disorders are multifactorial, including genetic and environmental factors.

[0003] Neurological disorders can be classified according to the primary symptoms affecting the patient. As further explained herein below, three main types of symptoms are observed: movement disorders, psychiatric (mood / social) disorders, and cognitive disorders.

[0004] Movement disorders include tremor, hypokinesia, e.g., bradykinesia or dyskinesia, muscle twisting, rigidity, postural instability, freezing of gait, etc. Diseases that present with movement disorders typically include Parkinson's disease, dystonia, epilepsy, Huntington's disease, and Tourette's syndrome.

[0005] Psychiatric disorders comprise a variety of illnesses that present with mood / social disturbance symptoms, including, but not limited to, autism spectrum disorders, schizophrenia disorders, bipolar disorders, depressive disorders, anxiety disorders, obsessive-compulsive disorders, substance-related disorders, and / or addictive disorders (as defined by the Diagnostic and Statistical Manual of Mental Disorders, 2013, fifth edition, the American Psychiatric Association). Some patients with movement disorders, such as Parkinson's disease and dystonia, may develop psychiatric disorders later in the course of their illness.

[0006] Cognitive impairment is present in many, if not all, mental disorders (e.g., schizophrenia, bipolar disorder). Only disorders whose fundamental feature is cognitive are included in the cognitive impairment category. Cognitive impairment affects the patient's daily life, making it difficult to accomplish simple tasks. Dementia is a typical cognitive impairment and is a general term for severe mental decline that interferes with daily life. Alzheimer's disease is a unique type of dementia accompanied by neurodegenerative aspects.

[0007] Neurological disorders are treated, when possible, with drugs that regulate neurotransmitter levels in the brain and their interactions with specific neurotransmitter receptors. The main neurotransmitters involved are glutamate, gamma-aminobutyric acid (GABA), dopamine, and acetylcholine. Glutamate and GABA neurotransmitters are of particular interest because of their key roles in increasing neuronal excitability (Platt et al., The Veterinary Journal, 2007, 173, 278-286: The role of glutamate in central nervous system health and disease - a review) and decreasing neuronal excitability (Holmes et al., Mental Retardation and Developmental Disabilities, 1995, 1, 208-219: Role of glutamate and GABA in the pathophysiology of epilepsy), respectively. Dopamine is involved in several brain functions. These include the basal ganglia-mediated control of movement (inadequate dopamine levels in the basal ganglia result in uncontrollable movements), pleasure-seeking behavior (which may lead to dysfunctional addictions), and cognition (which may lead to neurocognitive decline due to dopamine impairment in the frontal lobe) (Alcaro et al., Brain Res. ReV., 2007, 56(2), 283-321: Behavioral functions of the mesolimbic dopaminergic system: an affective neuroethological perspective). Acetylcholine is a neurotransmitter involved in learning and memory at the central nervous system level (Hasselmo et al., Curr Opin Neurobiol, 2006, 16(6), 710-715: The role of acetylcholine in learning and memory).

[0008] Levodopa is a common drug used to alleviate motor symptoms in Parkinson's disease. It is converted into dopamine in the brain, thus helping to balance dopamine deficiency. Levodopa is related to carbidopa, which helps prevent levodopa from being converted to dopamine throughout the body. One problem with levodopa treatment is the "on-off" phenomenon, which results in depression-related plateaus and periods of inability to perform tasks alternating with periods of jubilant thaw (Lees et al., J Neurology Neurosurgery Psychiatry, Special Supplement, 1989, 29-37: The on-off phenomenon). Lack of response to this treatment in patients with late-stage Parkinson's disease is a concern (Fabbri et al., Parkinsonism and related disorders, 2016: Do patients with late-stage Parkinson's disease still respond to levodopa?). Other common medications for treating symptoms of neuropsychiatric disorders, such as the "positive" symptoms, delusions and hallucinations in schizophrenia, are antipsychotics.

[0009] However, the therapeutic treatment of neuropathic symptoms with these drugs is non-specific and may induce serious adverse events. In addition, patients may become unresponsive to the drugs used.

[0010] As neuroscience advances, the brain can be thought of as an electrical network that encodes and transmits information through its electrical wires, the neurons. The connectivity between neurons is both simple and complex. Simple because the influx and outflow of ions within neurons generates action potentials (or "spikes" of electrical activity). Complex because the brain network is composed of hundreds of billions of neurons, which form nodes, hubs, and modules that exhibit coordinated interactions across various spatial and temporal scales (Fornito et al., Nature Reviews Neuroscience, 2015, 16, 159-172: The connectomics of brain disorders). Neurotransmission is determined by the anatomical components connecting individual neurons (structure) and the processes by which information is transmitted (function). Both aspects affect the overall performance of the nervous system. Neural interactions are mediated by oscillations in the brain's electrical activity patterns, which can typically be measured using an electroencephalogram (EEG). Different frequency bands of oscillations are observed: delta, theta, alpha, beta, and gamma (Ward et al., Trends in Cognitive Sciences, 2003, 7(12), 553-559: Synchronous neural oscillations and cognitive processes). Structurally, the brain's most striking neuroanatomical feature is the rich connectivity between neurons, reflecting the importance of neural communication. Synchrony of oscillations between one brain region and another ("synchrony") is thought to constitute the highest level of information encoding, resulting in spatiotemporal coordination: first level (neurons): action potentials; second level (neuronal networks): neuronal oscillations (Engel et al., Nature Reviews Neuroscience, 2001, 2, 704-716: Dynamic predictions: oscillations and synchrony in top-down processing).Importantly, there is emerging evidence that delicately balanced patterns of synchronization and desynchronization in space and time are fundamental to the functional performance of the nervous system (Schnitzler et al., Nature Reviews Neuroscience, 2005, 6, 285-296: Normal and pathological oscillatory communication in the brain).

[0011] Abnormal synchronization processes (too high and / or too long synchrony (i.e., also called hypersynchronization) or too low synchrony (i.e., also called dyssynchrony)) are associated with several brain disorders, such as epilepsy, schizophrenia, dementia, and Parkinson's disease (Schnitzler et al., Nature Reviews Neuroscience, 2005, 6, 285-296: Normal and pathological oscillatory communication in the brain).

[0012] Currently, modulation of neuronal electrical activity patterns (neuromodulation) can be induced by electrical stimulation. Current techniques for generating electrical stimulation in the brain utilize either direct electrical stimulation or induction of an electric field by applying current through a magnetic coil. Because certain neurological disorders affect deep brain regions and the penetration depth of electric fields is weak, surgical implantation of electrodes into the brain to deliver continuous electrical stimulation constitutes a "deep brain stimulation" (DBS) technique. Its effectiveness depends on the parameters used for stimulation, particularly the frequency. In 1987, high-frequency stimulation (≥100 Hz) of the ventral intermediate nucleus (VIM) with implanted electrodes was found to reduce tremor symptoms in patients with Parkinson's disease (Benabid et al., Applied Neurophysiology, 1987, 50, 344-346: Combined (thalamotomy and stimulation) stereotactic surgery of the VIM thalamic nucleus for bilateral Parkinson's disease). In monkeys, high-frequency stimulation (>100 Hz) compared with low-frequency stimulation (<50 Hz) has been shown to alter the temporal discharge patterns of neurons in the lateral pallidum (GPe) and medial pallidum (GPi) (a regular discharge pattern synchronized with the stimulation), which reduces bradykinesia and rigidity by blocking the transmission of altered patterns of neural activity in the basal ganglia to their target structures in the thalamus and brainstem (Hashimoto et al., The Journal of Neuroscience, 2003, 23(5), 1916-1923: Stimulation of the subthalamic nucleus changes the firing pattern of pallidal neurons). DBS is currently approved for treating several movement disorders (Parkinson's disease, dystonia, essential tremor, epilepsy) and psychiatric disorders (obsessive-compulsive disorder, depression).

[0013] However, several drawbacks may be associated with DBS. The first drawback is the invasiveness of the technique and the risk of various complications, such as bleeding, seizures, infection, lead migration, and lead breakage (Fenoy et al., J Neurosurg, 2014, 120, 132-139: Risks of common complications in DBS surgery: management and avoidance).

[0014] The focality of the electric field generated within the target (i.e., spatial resolution) is another concern. The spread of electrical stimulation has also been associated with side effects such as depression. Much research has been devoted to designing new types of electrodes that can shift the stimulation and confine it within a specific area (Luan et al., Frontiers in Neuroengineering, 2014, 7(27), 1-9: Neuromodulation: present and emerging methods). Other technical aspects are under evaluation: the electrodes (or leads), their size, the invasiveness of DBS devices, the materials that make up the leads, their compatibility with (magnetic resonance) imaging techniques, and the battery life of internal pulse generators (IPGs) related to the need for continuous stimulation.

[0015] The other major existing types of electrical stimulation, i.e., transcranial electrical stimulation or transcranial magnetic stimulation, have the advantage of being non-invasive, but the penetration depth of the electric field is weak. Therefore, their application is limited to stimulating the cerebral cortex (they cannot reach deep brain regions). Furthermore, their spatial resolution remains low.

[0016] The present invention features nanoparticles and / or nanoparticle aggregates (aggregates of nanoparticles) for use to / in the prevention or treatment of a neurological disorder (typically a disorder of a neuronal network) or at least one symptom thereof.

[0017] The nanoparticles or nanoparticle aggregates described herein by the inventors normalize (improve synchrony) the synchronization of neuronal oscillations within and / or between neuronal networks and within and / or between different brain regions, and thus help return a subject / patient to a healthy / normal state.

[0018] The nanoparticles and nanoparticle aggregates described herein by the inventors do not require the application / induction of an electric current or an electric field / electrical stimulation, and preferably do not require exposure to any other external activation source, e.g., a light source, a magnetic field, or an ultrasound source, in order to perform their function (i.e., be efficient). The nanoparticles and nanoparticle aggregates described herein do not require exposure to an electric current or an electric field / electrical stimulation, and preferably do not require exposure to any other external activation source, e.g., a light source, a magnetic field, or an ultrasound source, in order to be functional in the context of the uses described herein. The inventors have discovered that these nanoparticles or nanoparticle aggregates can be advantageously and surprisingly efficiently used without exposure to an electric current or an electric field / electrical stimulation, i.e., without exposing a subject to an electric current or an electric field / electrical stimulation, typically applied by, for example, deep brain stimulation (DBS), transcranial electrical stimulation (TES), or transcranial magnetic stimulation (TMS), and preferably without exposure to any other external activation source, e.g., a light source, a magnetic field, or an ultrasound source. This means that, according to the present invention, the subject being treated will not suffer from the negative side effects of exposure to electrical currents or fields / stimulation, or any other external activation source, such as a light source, a magnetic field or an ultrasound source.

[0019] As is well known to those skilled in the art, nanoparticles have an enhanced / high surface-to-volume ratio, with typically about 35% to 40% of atoms localized on the surface of a 10 nm nanoparticle, compared to less than 20% for nanoparticles larger than 30 nm. This high surface-to-volume ratio is associated with strong size-dependent surface reactivity. As a result, nanoparticles (especially nanoparticles smaller than 20 nm) may exhibit superior properties compared to bulk materials. For example, gold particles are known to be chemically inert and resistant to oxidation on a macroscopic scale, whereas gold particles smaller than 10 nm have chemically active surfaces. The toxicity mechanisms associated with the chemical destabilization of metallic nanoparticles could be (i) the direct release of metals in solution (dissolution process), (ii) the catalytic properties of metallic nanoparticles, and (iii) the development of redox properties on the nanoparticle surface, which can oxidize proteins, generate reactive oxygen species (ROS), and induce oxidative stress (see M. Auffan et al., Environmental Pollution 157 (2009) 1127-1133: Chemical Stability of metallic nanoparticles: a parameter controlling their potential cellular toxicity in vitro).

[0020] In addition to the gold nanoparticles described herein that exhibit catalytic properties, cerium oxide (7 nm CeO2 particles) or iron oxide (20 nm Fe3O4 particles) nanoparticles have also shown surface redox transformations that lead to oxidative stress-related cytotoxicity in vitro (see M. Auffan et al., Environmental Pollution 157 (2009) 1127-1133: Chemical Stability of Metallic Nanoparticles: A Parameter Controlling Their Potential Cellular Toxicity In Vitro). Similarly, 11 nm silica nanostructures are eroded by biological media (see SA Yang et al., Scientific Reports 2018 8:185: Silica nanoparticle stability in biological media revisited).

[0021] For this reason, as explained below by the inventors, nanoparticles having a size of less than 30 nm should be carefully selected if they are intended for in vivo use in a subject, typically a mammal, particularly a human.

[0022] Short description Herein, for the first time, the usefulness of nanoparticles or nanoparticle aggregates is described for use in preventing or treating a neurological disorder or at least one symptom thereof in a subject in need thereof, without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing the nanoparticles or nanoparticle aggregates to any other external activation source, such as a light source, a magnetic field, or an ultrasound source. The nanoparticle or nanoparticle aggregate material is typically a conductive material, a semiconducting material, a material with a dielectric constant ε of 200 or greater, and ijk Insulating materials with a dielectric constant ε of 100 or less ijk The insulating material is selected from the group consisting of:

[0023] In certain embodiments, the inventors herein describe nanoparticles or nanoparticle aggregates for use in the prevention or treatment of a neurological disorder or at least one symptom thereof in a subject, without exposing the nanoparticles or nanoparticle aggregates to an electric field or any other external activation source, the material of which the nanoparticles or nanoparticle aggregates are made being a conductive material, a semiconducting material, a material with a dielectric constant ε of 200 or greater, ijk Insulating materials with a dielectric constant ε of 100 or less ijk The insulating material is selected from an insulating material having the following structure: i) the maximum median core size of the nanoparticles or nanoparticle aggregates in the population is less than or equal to the maximum median core size of the nanoparticles or nanoparticle aggregates in the population if the material is a conductive material, a semiconductive material, or a material with a dielectric constant ε of 200 or more; ijk at least 30 nm when the insulating material has ii) The nanoparticle or nanoparticle aggregate core is coated with a biocompatible coating that provides a neutral or negative surface charge when measured in an aqueous solution having an electrolyte concentration of 0.001-0.2 M, a nanoparticle or nanoparticle aggregate material concentration of 0.01-10 g / L, and a pH of 6-8.

[0024] Also described herein is the use of nanoparticles or nanoparticle aggregates for the manufacture of a composition for preventing or treating a neurological disorder described herein or at least one symptom thereof in a subject in need thereof, without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing the nanoparticles or nanoparticle aggregates to any other external source of activation, such as a light source, a magnetic field, or an ultrasound source.

[0025] Also provided herein is a composition for use in preventing or treating a neurological disorder or at least one symptom thereof in a subject, the composition comprising or consisting of nanoparticles and / or nanoparticle aggregates and a pharmaceutically acceptable support, wherein the nanoparticle or nanoparticle aggregate material is typically a conductive material, a semiconductive material, a dielectric constant ε of 200 or greater, ijk Insulating materials with a dielectric constant ε of 100 or less ijkand wherein the prevention or treatment is performed without exposing the nanoparticles or nanoparticle aggregates administered to a subject via the composition to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field, or an ultrasound source.

[0026] Further, as used herein, there is provided a method for manufacturing a nanoparticle-based ... ijk Insulating materials with a dielectric constant ε of 100 or less ijk and its use in a method for preventing or treating a neurological disorder or at least one symptom thereof in a subject, typically without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing the nanoparticles or nanoparticle aggregates to any other external activation source, such as a light source, a magnetic field, or an ultrasound source.

[0027] Detailed Description The human nervous system is estimated to consist of approximately 80 to 120 billion nerve cells (Herculano-Houzel S. Frontier in Human Neuroscience (2009), 3(31): 1-11, The human brain in numbers: a linearly scaled-up primate brain). The defining property of a neuron (or nerve cell) is its ability to transmit electrical signals in the form of action potentials.

[0028] Neurons / nerve cells constitute the fundamental nodes of the brain. Neurons can communicate with each other in a highly structured manner to form neuronal networks. Neurons communicate via synaptic connections. Within neurons, nanocircuits constitute the underlying biochemical machinery for mediating important neuronal properties, such as learning and memory and the generation of neuronal periodicity.

[0029] Microcircuits can be formed by just a few interconnected neurons and can perform sophisticated tasks, such as mediating reflexes, processing sensory information, initiating movement, and mediating learning and memory. Macrocircuits are more complex networks consisting of multiple embedded microcircuits. Macrocircuits mediate higher brain functions, such as object recognition and cognition. Thus, multiple levels of networks populate the nervous system.

[0030] Excitability of neural networks Neurons transmit messages electrochemically (i.e., chemicals / ions create electrical signals). Important ions in the nervous system are sodium, potassium, calcium, and chloride. When a neuron is not transmitting signals, it is in a "resting state." When a neuron is resting, the inside of the neuron is negative relative to the outside. The concentrations of various ions attempt to balance on both sides of the membrane, but this is not possible because the cell membrane allows only some ions to pass through channels (ion channels). In addition to these selective ion channels, there are pumps that use energy to move three sodium ions out of the neuron for every two potassium ions that enter. Finally, when all these forces are balanced and the voltage difference between the inside and outside of the neuron is measured, the resting membrane potential (also known as "resting potential") of the neuron is approximately -70 mV. This means that the inside of the neuron is 70 mV lower than the outside. At rest, there are relatively more sodium ions outside the neuron and more potassium ions inside the neuron. An action potential (also identified as a "spike" or "impulse") occurs when a neuron leaves the cell body to send information down the axon. This means that some event (stimulus) moves the resting potential toward 0 mV. When depolarization reaches approximately -55 mV, the neuron discharges with an action potential. If the depolarization does not reach this critical threshold level, the neuron will not discharge with an action potential (on / off mechanism). Also, once the threshold level is reached, the neuron will always discharge with an action potential of a fixed magnitude. Thus, either the depolarization does not reach the threshold or a complete action potential is generated.

[0031] There is a large variability in the propagation speed of an action potential. In fact, the propagation speed of an action potential in a nerve can vary from 100 meters per second to less than a tenth of a meter per second. The time constant is a measure of how quickly the membrane will respond in time to a stimulus, while the spatial constant (also length constant) is a measure of how well the potential will spread along the axon as a function of distance.

[0032] Connectivity within and between neuronal networks There are three types of connectivity networks used to investigate communication within and across the brain. Structural connectivity is based on the detection of fiber structural pathways that physically connect brain regions. These are anatomical network maps that show the possible paths signals can travel through the brain. Functional connectivity identifies activity in brain regions with similar frequency, phase, and / or amplitude of correlated activity. Effective connectivity uses functional connectivity information and goes a step further to determine the direct or indirect influence one neural system can have on another, more specifically, the direction of dynamic information flow within the brain (Bowyer et al., Neuropsychiatric Electrophysiology, 2016, 2(1), 1-12: Coherence: a measure of the brain networks: past and present).

[0033] Synchronized activity within neuronal networks can be detected using magnetoencephalography (MEG), electroencephalography (EEG), functional magnetic resonance imaging (FMRI), or positron emission tomography (PET), followed by network connectivity analysis of the images. MEG or EEG are preferred because they have high temporal resolution for resolving the dynamic flow of information. Brain connectivity analysis is performed to map the communication networks necessary for the brain to function. Specific brain regions are specialized to process specific types of information. Imaging techniques have revealed that these regions connect and communicate with other specialized regions across brain networks. "Coherence" (Bowyer et al., Neuropsychiatric Electrophysiology, 2016, 2(1), 1-12: Coherence a measure of the brain networks: past and present) is a mathematical technique that quantifies the frequency and amplitude of the synchrony (synchronized or synchronized state) of neuronal patterns of oscillatory brain activity. The detection of synchronous neuronal activity can be used to determine the health or integrity of functional connectivity in the human brain. Overlaying functional connectivity maps onto structural connectivity images and using the direction of information flow derived from effective connectivity provides a comprehensive understanding of how the brain functions. These techniques are useful for evaluating therapeutic treatments based on pre- and post-treatment brain connectivity imaging.

[0034] The undamaged (i.e., "normal" or "healthy") brain exhibits complex patterns of synchronized activity associated with different "states" of the organism, ranging from slow delta (0.5-4 Hz) to theta (4-8 Hz), alpha (8-12 Hz), beta (15-30 Hz), and gamma (30-70 Hz) waves. Interestingly, isolated cultures of cortical structures provide a convenient system for examining the regularities that control the emergence, development, and spread of network discharges (spikes) and network bursts (groups of spikes) in densely interconnected populations of neurons. Network activity can be recorded noninvasively and over long periods with finite temporal resolution using multi-electrode arrays. Two-dimensional isolated cultures can be used as a viable test system to study the regularities that control the formation and maintenance of network activity in the brain, allowing the testing of hypotheses that cannot be addressed in the intact brain (Cohen E. et al., Brain Research, 2008, 1235, 21-30: Determinants of spontaneous activity in networks of cultured hippocampus).

[0035] Described herein for the first time are advantages of nanoparticles or nanoparticle aggregates for use in preventing or treating a neurological disorder or at least one symptom thereof in a subject in need thereof, without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing the nanoparticles or nanoparticle aggregates to any other external activation source, such as a light source, a magnetic field, or an ultrasound source. Such exposure to a (therapeutic or diagnostic) electric field, or to any other (therapeutic or diagnostic) external activation source, such as a light source, a magnetic field, or an ultrasound source, is typically understood herein to be a therapeutic or diagnostic exposure typically performed by medical staff, for example, by a doctor or nurse.

[0036] The material of the nanoparticles or nanoparticle aggregates is typically a conductive material, a semiconducting material, or a material with a dielectric constant ε of 200 or more. ijk Insulating materials with a dielectric constant ε of 100 or less ijk The insulating material is selected from the group consisting of:

[0037] The term "treatment" refers to a therapeutic procedure or measure that can prevent, alleviate, or cure a disease, disorder, or dysfunctional condition described herein. Such treatment is intended for mammalian subjects, preferably human subjects, in need of same. As such, subjects are contemplated who have already been identified (diagnosed) as suffering from a disease, disorder, or dysfunctional condition described herein, or who are considered to be "at risk of developing" such a disease, disorder, or dysfunctional condition for which the treatment is prophylactic or intended to be preventative.

[0038] In certain embodiments, the subject is not a subject suffering from epilepsy.

[0039] Abnormal modulation of oscillatory transmission between neurons is actually present in different types of neurological diseases or disorders (also identified herein as "neural diseases or disorders") (Uhlhaas et al., Neuron, 2006, 52, 155-168: Neural synchrony in brain disorders: relevance for cognitive dysfunctions and pathophysiology; Basar E. et al. International Journal of Psychophysiology 103 (2016) 135-148, What does the broken brain say to the neuroscientist? Oscillations and connectivity in schizophrenia, Alzheimer's disease, and bipolar disorder).

[0040] The human nervous system is divided into the central nervous system (CNS) and the peripheral nervous system (PNS). The CNS is in turn divided into the brain and spinal cord, which reside in the cranial cavity of the skull and the spinal canal, respectively. The CNS and PNS work in coordination to integrate sensory information and control motor and cognitive functions. Figure 1 shows a simplified diagram of the brain structure.

[0041] Synchronization within and / or between neuronal networks, and within and / or between different brain regions, is achieved temporally through the coordination of neuronal oscillations (Buzsaki et al., Science, 2004, 304, 1926-1929: Neuronal oscillations in cortical networks).

[0042] Movement disorders are typically due to hypersynchronization, which means that the synchronization of oscillations within and / or between neuronal networks within and / or between different regions of the brain, typically observed on an electroencephalogram (EEG), is too high and / or too long (excessive) when compared to healthy / normal subjects.

[0043] Psychiatric and cognitive disorders in subjects are typically due to synchronization disorders, meaning that the synchronization of oscillations within and / or between neuronal networks within and / or between different brain regions, typically observed in EEG, is reduced (typically showing reduced activity) or even absent, i.e., undetectable, when compared to healthy / normal subjects [see Table 1: Abnormal neural synchronization in neurological disorders (adapted from Uhlhaas et al., Neuron, 2006, 52, 155-168: Neural synchrony in brain disorders: relevance for cognitive dysfunctions and pathophysiology)].

[0044] [Table 1]

[0045] "Coherence" is a mathematical technique that quantifies the frequency and amplitude of synchrony (synchronized or synchronized states) in a subject's neuronal patterns of oscillatory brain activity, and therefore excessively high and low coherence compared to healthy / normal subjects can be considered to be involved in motor disorders and mental / cognitive disorders, respectively (Bowyer et al., Neuropsychiatric Electrophysiology, 2016, 2(1), 1-12: Coherence a measure of the brain networks: past and present) (see Figure 2).

[0046] In certain embodiments, the neurological disease or disorder targeted in the context of the present invention is selected from Parkinson's disease, Alzheimer's disease, epilepsy, obsessive-compulsive disorder, autism spectrum disorder, depressive disorder, dystonia, Tourette's syndrome, schizophrenia, stroke, aphasia, dementia, tinnitus, Huntington's disease, essential tremor, bipolar disorder, anxiety disorder, addiction disorder, vegetative state of consciousness, e.g., selected from Parkinson's disease, Alzheimer's disease, epilepsy, obsessive-compulsive disorder, autism spectrum disorder, depressive disorder, dystonia, Tourette's syndrome, schizophrenia, stroke, aphasia, dementia, tinnitus, Huntington's disease, essential tremor, bipolar disorder, addiction disorder, vegetative state of consciousness, and at least one symptom thereof.

[0047] As already explained herein above, neurological diseases or disorders can be classified according to the primary symptoms affecting the patient, which are movement disorders, psychiatric (mood / social) disorders and cognitive disorders, as further detailed herein below.

[0048] Examples of movement disorders Parkinson's disease Parkinson's disease (PD), which affects approximately 7 to 10 million people worldwide, is characterized by tremors, motor dysfunction, bradykinesia, and freezing of gait. PD is a slowly progressive degenerative brain disorder. PD affects neurons in the basal ganglia and substantia nigra, areas of the brain. Neurons in the substantia nigra produce dopamine, a neurotransmitter that acts as a chemical messenger in brain circuits important for the intention and control of physical movement. In PD, dopamine-producing neurons in the substantia nigra die prematurely in some individuals (Corti et al., Physiol Rev, 2011, 91, 1161-1218: What genetics tells us about the causes and mechanisms of Parkinson's disease). When dopamine receptors in the striatum are insufficiently stimulated, parts of the basal ganglia become either understimulated or overstimulated. In particular, the subthalamic nucleus (STN) becomes overactive and acts as a facilitator on the medial globus pallidus (GPi). Overstimulation of the GPi has an overinhibitory effect on the thalamus, which in turn reduces its output, resulting in motor slowing and rigidity (Guo et al., Frontiers in Computational Neuroscience, 2013, 7, 124, 1-11: Basal ganglia modulation of thalamocortical relay in Parkinson's disease and dystonia).

[0049] The lack of dopamine in PD is associated with excessive synchronization of oscillations at beta frequencies throughout the cortico-basal ganglia motor network. Indeed, dopamine levels in the basal ganglia are predicted to suppress beta synchrony, which in turn modulates the dopaminergic involvement required for motor preprocessing (Jenkinson et al., Trends in Neuroscience, 2011, 34(12), 611-618: New insights into the relationship between dopamine, beta oscillations, and motor function). When dopamine levels in the basal ganglia are not sufficiently high, the synchronization of beta oscillations is no longer controlled, and motor slowing may manifest. Other observations in patients with Parkinson's disease lead to the conclusion that cortical oscillations in the beta band initiate and drive beta oscillations in the basal ganglia (Lalo et al., The Journal of Neuroscience, 2008, 28(12), 3008-3016: Patterns of bidirectional communication between cortex and basal ganglia during movement in patients with Parkinson's disease).

[0050] Deep brain stimulation (DBS) can be used to treat symptoms of tremor and rigidity (Eusebio et al., J Neurol Neurosurg Psychiatry, 2011, 82, 569-573: Deep brain stimulation can suppress pathological synchronization in parkinsonian patients). DBS has been approved by the FDA for the treatment of PD symptoms since 2002 (for essential tremor since 1997). Electrical stimulation is typically performed in the basal ganglia, STN, and GPi. As mentioned above, cortical beta oscillations are also involved in the pathophysiology of the disease, so cortical transcranial stimulation (e.g., transcranial magnetic stimulation - TMS) can also be used to treat symptoms of Parkinson's disease (Cantello et al., Brain Research Reviews, 2002, 38, 309-327: Transcranial magnetic stimulation and Parkinson's disease).

[0051] dystonia Dystonia is a neurological disorder characterized by abnormal, involuntary twisting and rotating movements that reflect impaired motor system function. Several forms of dystonia exist, depending on the affected body part, their genetic origin, and the type of neurotransmitter involved. In dystonia, central nervous system (CNS) inhibition is incomplete, resulting in a loss of reciprocal spinal inhibition between opposing muscles. For example, in upper limb dystonia, abnormal synchronization of neurons / nerves providing input signals to antagonistic muscles of the forearm results in co-contraction of these antagonistic muscles (the symptom of dystonia) (Farmer et al., Brain, 1998, 121, 801-814: Abnormal motor unit synchronization of antagonist muscles underlies pathological co-contraction in upper limb dystonia).

[0052] A DBS target site showing interesting antidystonic effects is the medial globus pallidus (GPi-DBS). GPi-DBS was approved by the FDA in 2003 for patients with chronic, medically refractory dystonia (Hu et al., Translational Neurodegeneration, 2014, 3(2), 1-5: Deep brain stimulation for dystonia). Stimulation of the ventral intermedius (VIM) nucleus of the thalamus (VIM-DBS) has a much weaker effect. Stimulation using the subthalamic nucleus (STN-DBS) has been experimental. GPi-DBS reduces the core symptoms of dystonia, but the full therapeutic effect may take weeks to months to manifest (Dressler et al., J Neural Transm, 2015, DOI 10.1007 / s00702-015-1453-x: Strategies for treatment of dystonia).

[0053] epilepsy Epilepsy is a brain disorder that affects approximately 50 million people worldwide and is primarily characterized by recurrent and unpredictable interruptions of normal brain function, called epileptic seizures. Epilepsy is not a single disease entity, but a spectrum of disorders reflecting underlying brain dysfunction that may result from many different causes (genetic mutations, brain tumors, head trauma, stroke, alcoholism, brain inflammation; infections such as meningitis, HIV, or viral encephalitis) (Fisher et al., Neurology, 2015, 28(2), 130-135: Redefining epilepsy). An epileptic seizure is defined as a transient occurrence of signs and / or symptoms due to excessive synchronous neuronal activity in the brain (Fisher et al., Epilepsia, 2005, 46(4), 470-472: Epileptic seizures and epilepsy: definitions proposed by the International League Against Epilepsy (ILAE) and the International Bureau for Epilepsy (IBE)). The cerebral cortex is a major component in the development of epileptic seizures, and many people are diagnosed with focal frontal lobe seizures or mesial temporal lobe seizures (National Institute of Neurological Disorders and Stroke: http: / / www.ninds.nih.gov / disorders / epilepsy / detail_epilepsy.htm#3109_7). The identification of areas of elevated local synchronization or "hypersynchrony" in the cortex suggests that local hypersynchrony may be a marker of areas of seizure onset (Schevon et al., Neuroimage, 2007, 35(1), 140-148: Cortical abnormalities in epilepsy revealed by local EEG synchrony).

[0054] Neurostimulation for the treatment of epilepsy can take the form of peripheral nerve stimulation, such as vagus nerve stimulation (VNS), spinal cord stimulation, transcranial brain stimulation (TES or TMS), or deep brain stimulation (DBS). Responsive neurostimulation is another strategy. In this case, stimulation is performed only when seizure onset is detected. Both VNS and responsive neurostimulation are approved by the FDA in the United States for the treatment of certain types of epilepsy. DBS of the anterior thalamic nucleus (ANT) is approved in European Union countries (Fisher et al., Nature Reviews Neurology, 2014, 10, 261-270: Electrical brain stimulation for epilepsy).

[0055] Examples of mental disorders (mental / social disorders) Obsessive-Compulsive Disorder (OCD) Obsessive-compulsive disorder (OCD) is a common psychiatric disorder that is often chronic, severe, and extremely debilitating, and is typically unresponsive to treatment, with a significant proportion of patients either not responding or only achieving partial relief.

[0056] Functional neuroimaging studies have demonstrated dysfunction in the orbitofrontal cortex, basal ganglia, and striatum.

[0057] One study showed that acute OCD symptoms may be associated with abnormal high-oscillatory activity in the subthalamic nucleus (STN), particularly in the left hemisphere and in the delta-alpha (1-12 Hz) frequency range (Bastin et al., Cortex, 2014, 60, 145-150: Changes of oscillatory activity in the subthalamic nucleus during obsessive-compulsive disorder symptoms: two case reports). Furthermore, some subthalamic neurons specifically increased their discharge rate when doubts arose during a checking task (Burbaud et al., Brain, 2013, 136(1), 304-317: Neuronal activity correlated with checking behavior in the subthalamic nucleus of patients with obsessive-compulsive disorder).

[0058] DBS of the ventral anterior limb of the internal capsule (VC) and the adjacent ventral striatum (VS) has been approved in the EU for the treatment of severe and highly treatment-resistant OCD (VC / VS-DBS).

[0059] Autism spectrum disorder Autism is a neurodevelopmental syndrome defined by deficits in social interaction and communication and abnormally restricted and repetitive behaviors. It is a disorder that typically begins in infancy, or at the latest within the first three years of life. Autism is a heterogeneous condition (no two children or adults with autism have the same profile), which has led to the concept of "autism spectrum disorders," which categorize the disorder into several levels according to the degree of language deficit or global cognitive delay and the severity of social or behavioral symptoms (Lord et al., Neuron, 2000, 28, 355-363: Autism spectrum disorders). At one end of the spectrum, individuals with autism are high-functioning and able to live independently and maintain employment. Individuals characterized as low-functioning exhibit more severe symptoms, namely, difficulties with language (and nonverbal language), poor social communication, self-injurious behaviors (SIB), tantrums, and potentially life-threatening aggression. The involvement of networks for socio-emotional processing, namely the limbic system, face processing system and mirror neuron network, has become an important trend in structural and functional brain research in autism. Deficits in gamma-band oscillation synchrony have been shown to be involved in the emergence of symptoms (Sinha et al., Neurosurgery Focus, 2015, 38(6), E3: Deep brain stimulation for severe autism: from pathophysiology to procedure).

[0060] Two major symptom areas that may require treatment in severe autism are social deficits, including nonverbality and unresponsiveness to conversation, and potentially life-threatening social inactivity (SIB). The amygdala is thought to play a key role in these abnormal pathophysiologies. Alterations in excitatory or inhibitory control imply abnormalities in the pathophysiology of autism. Neuromodulation targeting the amygdala with DBS represents a therapeutic intervention for patients with severe autism. Three cases of DBS treatment have been reported in the literature. The goal of treatment was primarily to reduce disease-related motor disorders, such as stereotypic behaviors (repetitive movement patterns) and self-injurious behaviors (SIB) (Sinha et al., Neurosurgery Focus, 2015, 38(6), E3: Deep brain stimulation for severe autism: from pathophysiology to procedure; Stocco et al., Parkinsonism and related disorders, 2014, 20, 1035-1036: Deep brain stimulation for severe secondary stereotypies). In one of three cases, DBS in the lateral basal ganglia was reported to significantly improve autism-related symptoms, such as social interaction and affect mood and nighttime sleep (Sturm et al., Frontiers in Human Neuroscience, 2013, 6, 341, 1-10).

[0061] Schizophrenia Schizophrenia is a chronic mental illness characterized, inter alia, by the following symptoms: positive symptoms (hallucinations and delusions) that reflect abnormal mental activity; negative symptoms (disordered thought, blunted affect, speech difficulties) that correspond to deficits in normally present mental functioning. As a cause of lifetime disability, schizophrenia is among the top 10.

[0062] Marked ventricular enlargement and increased cerebrospinal fluid on the brain surface suggest that the brain is shrinking. This loss of gray matter and the reduction in the number of synaptic structures on neurons suggests that schizophrenia is a neurodevelopmental disorder, meaning that brain abnormalities (as opposed to neurodegenerative disorders) are already present in patients with early symptoms.

[0063] It has been demonstrated that the observed neural circuit impairments in schizophrenic patients result from dysfunction of gamma-band synchronization (Spencer et al., The Journal of Neuroscience, 2003, 23(19), 7407-7411: Abnormal neural synchrony in schizophrenia; Gallinat et al., Clinical Neurophysiology, 2004, 115, 1863-1874: Reduced oscillatory gamma-band responses in unmedicated schizophrenic patients indicate impaired frontal network processing).

[0064] Electroconvulsive therapy (ECT), or shock treatment, has proven to be one of the most successful non-pharmacological treatments for schizophrenia (Payne et al., J. Psychiatr. Pract., 2009, 15(5), 346-368: Electroconvulsive therapy part I: a perspective on the evolution and current practice of ECT). ECT involves the continuous application of an electric current to the brain, which induces seizures comparable to epileptic seizures.

[0065] DBS can also be used for the symptomatic treatment of schizophrenia. For example, DBS of the nucleus accumbens (NAcc) in depressed patients tends to alleviate anhedonia, i.e., restores pleasure (Schlaepfer et al., Neuropsychopharmacology, 2008, 33, 368-377: Deep brain stimulation to reward circuitry alleviates anhedonia in refractory major depression).

[0066] Examples of cognitive impairment Alzheimer's disease Alzheimer's disease (AD) is a neurodegenerative disorder that causes mental, behavioral, and functional decline and progressive loss of learning ability. As of 2013, an estimated 5.2 million Americans had AD, with approximately 200,000 under the age of 65 and 5 million over the age of 65 (Alzheimers Dement. 2013, 9(2), 208-245: 2013 Alzheimer's disease facts and figures).

[0067] Recent evidence indicates that the cognitive impairment seen in Alzheimer's disease is related to functional disconnection of neurocognitive networks. Analysis of global EEG synchronization reveals widespread decreases in alpha, beta, and gamma band synchronization, accompanied by increases in delta band synchronization. In patients with mild Alzheimer's disease, loss of beta band synchronization has been shown to correlate with cognitive impairment (Schnitzler et al., Nature Reviews Neuroscience, 2005, 6, 285-296: Normal and pathological oscillatory communication in the brain). Clinical studies are underway to evaluate the potential of DBS for the treatment of Alzheimer's disease.

[0068] nanoparticles Described herein are nanoparticles or nanoparticle aggregates for use in accordance with the present invention for / in the prevention or treatment of a neurological disorder or at least one symptom thereof in a subject, without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field, or an ultrasound source, wherein the material of the nanoparticles or nanoparticle aggregates is typically a conductive material, a semiconducting material, a material with a dielectric constant ε of 200 or greater, or ijk Insulating materials with a dielectric constant ε of 100 or less ijk The insulating material is selected from the group consisting of:

[0069] Dimensions or size of nanoparticles or nanoparticle aggregates For the purposes of the present invention, the term "nanoparticle" or "nanoparticle aggregate" refers to a product, in particular a synthetic product, having a size in the nanometer range, typically 1 nm to 1000 nm or 1 nm to 500 nm, e.g., at least 10 nm to about 500 nm or about 1000 nm, at least 30 nm to about 500 nm or about 1000 nm, at least 40 nm to about 500 nm or about 1000 nm, at least 45 nm to about 500 nm or about 1000 nm, preferably less than 500 nm.

[0070] The terms "nanoparticle aggregate" or "nanoparticle aggregate" refer to a collection of nanoparticles that are strongly bound, typically covalently bonded, to one another.

[0071] Electron microscopy, such as scanning electron microscopy (SEM), transmission electron microscopy (TEM), or cryo-TEM, can be used to measure the size of nanoparticles or nanoparticle aggregates, more specifically, the cores of nanoparticles or nanoparticle aggregates, i.e., the cores of nanoparticles or nanoparticle aggregates without their biocompatible coatings. In practice, biocompatible coatings are typically prepared from compounds (polymers or organic compounds) primarily composed of light components, which exhibit relatively weak elastic interactions with energetic electrons, resulting in low image contrast. TEM measures the projection images of particles deposited on an electron-transparent substrate. Typically, more than about 50 nanoparticles or nanoparticle aggregates per sample should be measured for size assessment. Therefore, more than about 50 nanoparticles or nanoparticle aggregates, or preferably more than about 100, 150, or 200 nanoparticles or nanoparticle aggregates, can be used to determine the maximum median core size of the nanoparticles or nanoparticle aggregates in the population, as well as the core size of nanoparticles or nanoparticle aggregates representing 30% to 70% of the population of nanoparticles and nanoparticle aggregates. A typical assay protocol can be found in "NIST - NCL Joint Assay Protocol, PCC-7; Measuring the size of using transmission electron microscopy (TEM); version 1.1 December 2009."

[0072] Similarly, dynamic light scattering (DLS) can be used to measure the hydrodynamic diameter of nanoparticles or nanoparticle aggregates in solution (i.e., the diameter of the nanoparticle or nanoparticle aggregate, including both its core and its biocompatible coating). The hydrodynamic diameter is the diameter of an equivalent hard sphere that diffuses at the same rate as the analyte. A typical assay protocol can be found in "NIST - NCL Joint Assay Protocol, PCC-1; Measuring the size of nanoparticles in aqueous media using batch-mode dynamic light scattering; version 1.1 February 2010." Particle size results obtained from DLS measurements may not be consistent with results obtained from other techniques (e.g., electron microscopy). This is in part due to differences in the actual physical properties measured (e.g., hydrodynamic diffusion vs. projected area). Furthermore, DLS is sensitive to the presence of small amounts of large or small particle aggregates. On the other hand, electron microscopy typically reflects the size of the primary particles (i.e., the core size of nanoparticles or nanoparticle aggregates) (see NIST - NCL Joint Assay Protocol, PCC-1; Measuring the size of nanoparticles in aqueous media using batch-mode dynamic light scattering; version 1.1 February 2010).

[0073] These two methods, DLS and electron microscopy, can be used in turn to further compare size measurements and confirm the size. The preferred method for measuring the size of nanoparticles and nanoparticle aggregates is DLS (see International Standard ISO22412 Particle Size Analysis - Dynamic Light Scattering, International Organization for Standardization (ISO) 2008). The mean hydrodynamic diameter of nanoparticles or nanoparticle aggregates measured by DLS in solution is expressed as an intensity size distribution (light scattering intensity is proportional to particle size) and is measured at room temperature (approximately 25°C).

[0074] Typically, the largest dimension or size is the diameter of a round or spherical nanoparticle or the longest dimension of an oval or elliptical nanoparticle.

[0075] The maximum dimension of the nanoparticles or aggregates defined herein is typically from about 2 nm to about 250 nm or about 500 nm, preferably from about 4 nm or 10 nm to about 100 nm or about 200 nm, even more preferably from about (preferably at least) 10 nm to about 150 nm, from about (preferably at least) 30 nm to about 150 nm, from about (preferably at least) 40 nm to about 500 nm, from about (preferably at least) 45 nm to about 500 nm, preferably less than 500 nm.

[0076] DLS techniques are typically used to measure the average hydrodynamic diameter of nanoparticles or nanoparticle aggregates in solution. Using DLS, the average hydrodynamic diameter of nanoparticles or nanoparticle aggregates in solution is typically about 10 nm to about 500 nm, preferably about 10 nm or about 30 nm to about 100 nm or about 500 nm, and even more preferably about 10 nm or about 30 nm to about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, or about 500 nm.

[0077] When the cores of nanoparticles or nanoparticle aggregates are measured, electron microscopy techniques are typically used. Using electron microscopy, the median maximum size (also identified herein as the "median maximum dimension") of the cores of the nanoparticles or nanoparticle aggregates of the population is typically about 5 nm to about 250 nm or about 500 nm, preferably about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, about 14 nm, about 15 nm, about 16 nm, about 17 nm, about 18 nm, about 19 nm, About 20 nm, about 21 nm, about 22 nm, about 23 nm, about 24 nm, about 25 nm, about 26 nm, about 27 nm, about 28 nm, about 29 nm, about 30 nm, about 31 nm, about 32 nm, about 33 nm, about 34 nm, about 35 nm, about 36 nm, about 37 nm, about 38 nm, about 39 nm, about 40 nm, about 41 nm, about 42 nm, about 43 nm, about 44 nm or about 45 nm to about 75 nm, about 76 nm, about 77 nm, about 78 nm, about 79 nm, about 80nm, about 81nm, about 82nm, about 83nm, about 84nm, about 85nm, about 86nm, about 87nm, about 88nm, about 89nm, about 90nm, about 91nm, about 92nm, about 93nm, about 94nm, about 95n m, about 96nm, about 97nm, about 98nm, about 99nm, about 100nm, about 101nm, about 102nm, about 103nm, about 104nm, about 105nm, about 106nm, about 107nm, about 108nm, about 109n m, approx. 110 nm, approx. 111 nm, approx. 112 nm, approx. 113 nm, approx. 114 nm, approx. 115 nm, approx. 116 nm, approx. 117 nm, approx. 123nm, about 124nm, about 125nm, about 130nm, about 140nm, about 150nm, about 200nm, about 250nm, about 300nm, about 350nm, about 400nm, about 450nm or about 500nm.

[0078] Typically, when electron microscopy tools are used to measure the core size of nanoparticles or nanoparticle aggregates, the core sizes of nanoparticles or nanoparticle aggregates representing 30% to 70% of the population of nanoparticles and nanoparticle aggregates are about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, about 14 nm, about 15 nm, about 16 nm, about 17 nm, about 18 nm, about 19 nm, about 20 nm, and the like. , about 21 nm, about 22 nm, about 23 nm, about 24 nm, about 25 nm, about 26 nm, about 27 nm, about 28 nm, about 29 nm, about 30 nm, about 31 nm, about 32 nm, about 33 nm, about 34 nm, about 35 nm, about 36 nm, about 37 nm, about 38 nm, about 39 nm, about 40 nm, about 41 nm, about 42 nm, about 43 nm, about 44 nm or about 45 nm to about 75 nm, about 76 nm, about 77 nm, about 78 nm, about 79 nm, about 80 nm, about 81nm, about 82nm, about 83nm, about 84nm, about 85nm, about 86nm, about 87nm, about 88nm, about 89nm, about 90nm, about 91nm, about 92nm, about 93nm, about 94nm, about 95nm, about 96nm , about 97nm, about 98nm, about 99nm, about 100nm, about 101nm, about 102nm, about 103nm, about 104nm, about 105nm, about 106nm, about 107nm, about 108nm, about 109nm, about 110nm , about 111 nm, about 112 nm, about 113 nm, about 114 nm, about 115 nm, about 116 nm, about 117 nm, about 118 nm, about 119 nm, about 120 nm, about 121 nm, about 122 nm, about 123 nm, about 124 nm, about 125 nm, about 130 nm, about 140 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm or about 520 nm.

[0079] Nanoparticle composition Nanoparticles prepared from conductive materials The nanoparticles prepared from conductive materials may be organic or inorganic nanoparticles.

[0080] Inorganic nanoparticles prepared from conductive materials typically have E° values ​​more positive than that of the standard hydrogen electrode, as measured at 25°C and 1 atm pressure ("Reduction reactions having E° values ​​more positive than that of the standard hydrogen electrode," Handbook of chemistry and physics; David R. Lide; 88 th The nanoparticles are prepared using a metal element having a standard reduction potential E° value of about 0.01 or more, more preferably about 0.1, 0.2, 0.3, 0.4, or 0.5 or more (see Table 2 of the International Edition). Representative metal elements used to prepare the nanoparticles can be selected from Tl, Po, Ag, Pd, Ir, Pt, Au, and mixtures thereof. Preferably, the metal element usable as a conductor material for preparing the nanoparticles is selected from Ir, Pd, Pt, Au, and mixtures thereof, and even more preferably, Au, Pt, Pd, and any mixtures thereof. Particularly preferred materials are Au and Pt.

[0081] Typically, gold nanoparticles have shown catalytic activity when their size is reduced to a few nanometers (see M. Auffan et al., Nature Nanotechnology 2009, 4(10), 634-641: Towards a definition of inorganic nanoparticles from an environmental, health and safety perspective). In order to minimize the surface contribution of inorganic nanoparticles to catalytic activity by reducing the surface / volume ratio, it is preferred that the maximum median core size of the nanoparticles or nanoparticle aggregates of the population is at least 30 nm, typically at least 40 nm or at least 45 nm. Interestingly, the inventors have shown that gold nanoparticles with a maximum median core size of the nanoparticles or nanoparticle aggregates of the population equal to 45 nm and / or with nanoparticle or nanoparticle aggregate core sizes of 42 nm to 49 nm that represent 30% to 70% of the population of nanoparticles and nanoparticle aggregates have a greater effect on MPP activity on neuronal networks than gold nanoparticles with a maximum median core size of the nanoparticles or nanoparticle aggregates of the population equal to 15 nm and / or with nanoparticle or nanoparticle aggregate core sizes of 14 nm to 16 nm that represent 30% to 70% of the population of nanoparticles and nanoparticle aggregates, even though the gold nanoparticles tested contain the same gold concentration (see Examples 9 and 10). + Prevents induced functional effects / of the MPP + We found that acetaminophen was more effective in relieving induced functional effects.

[0082] Organic nanoparticles prepared from conductive materials are typically prepared from organic materials having consecutive sp2 hybridized carbon centers in their structure (i.e., carbon-carbon double bonds or aromatic rings containing heteroatoms, typically N or S, within or outside the aromatic ring). Preferred organic materials are selected from polyaniline, polypyrrole, polyacetylene, polythiophene, polycarbazole, propylene, poly(3,4-ethylenedioxythiophene) and / or poly(3,4-ethylenedioxythiophene) polystyrene sulfonate.

[0083] In a particular embodiment, when the material is a conductive material as described herein above, the maximum median core size of the nanoparticles or nanoparticle aggregates of the population is at least 30 nm or at least 40 nm, as described herein above, preferably less than 500 nm, e.g., 45 nm; said conductive material is in particular a metallic material, typically a metal having a standard reduction potential E° of more than 0.2, or an organic material, typically an organic material having consecutive sp2 hybridized carbon centers in its structure, preferably a metallic material as described herein above, in particular any one of Au, Pt, Pd and any mixture thereof.

[0084] Nanoparticles prepared from semiconductor materials Nanoparticles prepared from semiconductor materials are typically inorganic nanoparticles.

[0085] Inorganic nanoparticles are typically prepared from semiconductor materials that exhibit a relatively small energy band gap (Eg) between their valence and conduction bands. Typically, semiconductor materials have a band gap Eg of less than 3.0 eV, typically measured at room temperature (about 25°C) (see, e.g., Handbook of chemistry and physics; David R. Lide; 88 th (See Tables 12-77, Table 3 of the IEEE International Symposium on Materials Science and Technology, Vol. 1, No. 1, pp. 111-115, 1997.) In certain embodiments, the material is an intrinsic or extrinsic semiconductor material, as further described herein below.

[0086] Intrinsic semiconductor materials typically consist of elements from Group IVA of the Mendeleev periodic table, such as silicon (Si) or germanium (Ge), mixed compositions of elements from Groups III and V of the Mendeleev periodic table, such as AlSb, AlN, GaP, GaN, InP, InN, etc., or mixed compositions of elements from Groups II and VI of the Mendeleev periodic table, such as ZnSe, ZnTe, CdTe, etc.

[0087] Extrinsic semiconductor materials typically comprise or consist of intrinsic semiconductors prepared with a high degree of chemical purity, where the intrinsic semiconductor material includes a dopant. In certain embodiments, when the extrinsic semiconductor material of the nanoparticles or nanoparticle aggregates consists of an element from Group IV A of the Mendeleev periodic table, it is doped with charge carriers selected from Al, B, Ga, In, and P. Such extrinsic semiconductor materials can be either n-type, where negative charge carriers predominate, or p-type, where positive charge carriers predominate. Typical extrinsic p-type semiconductor materials consist of silicon (Si) or germanium (Ge) doped with charge carriers selected from aluminum (Al), boron (B), gallium (Ga), and indium (In). Typical extrinsic p-type semiconductor materials consist of silicon (Si) or germanium (Ge), typically doped with phosphorus (P).

[0088] Typically, it has been shown that the band gap energy of semiconductor nanoparticles increases when the size of the nanoparticles is reduced below 10 nm (see M. Auffan et al., Nature Nanotechnology 2009, 4(10), 634-641: Towards a definition of inorganic nanoparticles from an environmental, health and safety perspective). In order to ensure a low surface / volume ratio and maintain the bulk band gap of the nanoparticles or nanoparticle aggregates below 3.0 eV, it is preferred that the maximum median core size of the nanoparticles or nanoparticle aggregates of the population is at least 30 nm, preferably at least 40 nm.

[0089] Thus, in certain embodiments, the maximum median core size of the nanoparticles or nanoparticle aggregates of the population is at least 30 nm or at least 40 nm, preferably less than 500 nm, when the material is a semiconductor material as described herein above, in particular a semiconductor material having a band gap Eg of less than 3.0 eV, typically consisting of an element from group IV A of the Mendeleev periodic table, in particular an element from group IV A of the Mendeleev periodic table doped with charge carriers selected from Al, B, Ga, In and P, or a mixed composition of elements from groups III and V of the Mendeleev periodic table or a mixed composition of elements from groups II and VI of the Mendeleev periodic table.

[0090] Nanoparticles prepared from insulating materials with a high relative permittivity, i.e., a relative permittivity of 200 or more. High relative permittivity ε ijk Nanoparticles prepared from or consisting of insulating materials typically have a band gap Eg of 3.0 eV or greater when typically measured at room temperature (about 25°C) and a dielectric constant (also called a dielectric constant) of 1.0 eV or greater when measured at 20°C to 30°C and 10°C. 2 Typically measured at frequencies between 1000 and 1000 Hz (see, e.g., "Permittivity (dielectric constant) of inorganic solids"; Handbook of chemistry and physics; David R. Lide; 88 th (See Tables 12-45 in "Compilation of the static dielectric constant of inorganic solids," K.F. Young and H.P.R. Frederikse, J. Phys. Chem. Ref. Data, Vol. 2, No. 2, 1973) Relative permittivity ε of 200 or more ijk and prepared from a material having the following properties.

[0091] Such nanoparticles are prepared with a dielectric material that is typically a mixed metal oxide, preferably selected from BaTiO3, PbTiO3, KTaNbO3, KTaO3, SrTiO3, BaSrTiO3, and the like.

[0092] Typically, PbTiO nanoparticles, which are perovskite-based structures, exhibit a change in their paraelectric to ferroelectric transition temperature for nanoparticle sizes below 20-30 nm (see M. Auffan et al., Nature Nanotechnology 2009, 4(10), 634-641: Towards a definition of inorganic nanoparticles from an environmental, health, and safety perspective). To ensure a low surface-to-volume ratio and maintain the dielectric properties of the nanoparticles or nanoparticle aggregates, the maximum median core size of the nanoparticles or nanoparticle aggregates in the population is preferably at least 30 nm, typically at least 40 nm.

[0093] Thus, in certain embodiments, the maximum median core size of the nanoparticles or nanoparticle aggregates in the population is such that the material has a high dielectric constant ε of 200 or greater. ijk In the case of the insulating materials described herein above, in particular insulating materials having a band gap Eg of 3.0 eV or greater, preferably mixed metal oxides selected from BaTiO3, KTaNbO3, KTaO3, SrTiO3 and BaSrTiO3, the thickness is at least 30 nm or at least 40 nm, preferably less than 500 nm.

[0094] Nanoparticles prepared from insulating materials with a low dielectric constant (dielectric constant), i.e., a dielectric constant of 100 or less. Nanoparticles prepared from or consisting of insulating materials with low dielectric constants typically have a band gap Eg of 3.0 eV or more when measured at room temperature (25°C) and a band gap Eg of 1.0 eV or more when measured at 20°C to 30°C and 10°C. 2Hz to infrared frequencies (see, for example, "Permittivity (dielectric constant) of inorganic solids"; Handbook of chemistry and physics; David R. Lide; 88 th Edition; Compilation of the static dielectric constant of inorganic solids. K.F. Young and H.P.R. Frederikse. J. Phys. Chem. Ref. Data, Vol. 2, No. 2, 1973, see Tables 12-45) of 100 or less, preferably 50 or less or 20 or less. ijk Typically, the material is prepared from a material having the following properties:

[0095] Such nanoparticles are typically prepared using a dielectric material selected from metal oxides, mixed metal oxides (wherein the metal element of the compound is derived from a metal element from period 3, 5, or 6 of the Mendeleev periodic table or a lanthanide), and carbon materials. The dielectric material is preferably selected from Al2O3, LaAlO3, La2O3, SiO2, SnO2, Ta2O5, ReO2, ZrO2, HfO2, and carbon diamond. More preferably, the dielectric material is a metal oxide selected from ReO2, ZrO2, HfO2, and any mixture thereof. Dielectric materials selected from ZrO2 and HfO2 are particularly preferred. In certain preferred embodiments, the dielectric material or metal oxide is not CeO2 (cerium oxide), Fe3O4 (iron oxide), SiO2 (silica), or any mixture thereof.

[0096] Both zirconium (Zr) and hafnium (Hf) are + In the oxidation state, Zr 4+ and Hf 4+The elements Zr4+ and Hf4+ are nearly identical in size and chemical properties. For this reason, these two ions are considered together in establishing their aqueous chemistry (see chapter 8, section 8.2 Zr4+ and Hf4+, p. 147 "The hydrolysis of cations," Baes CF & Mesmer RE; John Wiley and Sons, Inc. reprint Edition 1986).

[0097] In certain embodiments, the maximum median core size of the nanoparticles or nanoparticle aggregates of the population is at least 10 nm, preferably less than 500 nm, as defined herein above, when the material is selected from ReO2, ZrO2, HfO2, preferably ZrO2 and HfO2, and any mixture thereof, as defined herein above.

[0098] Shape of nanoparticles or nanoparticle aggregates The shape of the particles or aggregates can affect their "biocompatibility", so particles or aggregates with a very uniform shape are preferred. For pharmacokinetic reasons, therefore, nanoparticles or aggregates that are essentially spherical, round or ovoid in shape are preferred. Such shapes also favor the interaction of the nanoparticles or aggregates with or uptake by cells. Spherical or round shapes are particularly preferred.

[0099] The shape of nanoparticles or nanoparticle aggregates is typically assessed using electron microscopy, for example, transmission electron microscopy (TEM).

[0100] Biocompatible coatings of nanoparticles or nanoparticle aggregates In a preferred embodiment, the core of the nanoparticles or nanoparticle aggregates used in the context of the present invention to prepare the composition of interest can be coated with a biocompatible material selected from agents exhibiting stealth properties. The agent exhibiting stealth properties can be an agent exhibiting a steric group. Such groups can be selected from, for example, polyacrylate; polyacrylamide (poly(N-isopropylacrylamide)); polycarbamide; biopolymers; polysaccharides, such as dextran or xylan; and collagen. In another preferred embodiment, the core of the nanoparticles or nanoparticle aggregates can be coated with a biocompatible material selected from agents that enable interaction with biological targets. Such agents can typically impart a positive or negative charge to the surface of the nanoparticles or nanoparticle aggregates. An agent that forms a positive charge on the surface of the nanoparticles or nanoparticle aggregates can be, for example, aminopropyltriethoxysilane or polylysine. The agent that forms a negative charge on the surface of the nanoparticles or nanoparticle aggregates can be, for example, a phosphate (e.g., polyphosphate, metaphosphate, pyrophosphate, etc.), a carboxylate (e.g., citrate or a dicarboxylic acid, particularly succinic acid), or a sulfate.

[0101] In a preferred embodiment, the core of the nanoparticles or nanoparticle aggregates used in the context of the present invention is coated with a biocompatible material (i.e., coating agent) that confers a hydrophilic neutral surface charge or is selected from hydrophilic agents that impart a neutral surface charge to the nanoparticles. Indeed, when the nanoparticles of the present invention are administered to a subject, nanoparticles that confers a hydrophilic neutral surface charge or nanoparticle cores coated with a biocompatible agent selected from hydrophilic agents that impart a neutral surface charge to the nanoparticles are particularly advantageous for optimizing the use of the nanoparticles described herein to treat neurological disorders.

[0102] The hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticles or nanoparticle aggregates can be an agent exhibiting a functional group selected from alcohol (R—OH), aldehyde (R—COH), ketone (R—CO—R), ester (R—COOR), acid (R—COOH), thiol (R—SH), sugar (e.g., glucose, fructose, ribose), acid anhydride (RCOOOC-R), and pyrrole. The hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticles or nanoparticle aggregates can be a monomer, dimer, oligomer, polymer, or copolymer. When the agent is an oligomer, the oligomer can be an oligosaccharide, such as cyclodextrin. When the agent is a polymer, the polymer can be a polyester (e.g., poly(lactic acid) or polyhydroxyalkanoic acid), polyether, polyethylene oxide, polyethylene glycol, polyvinyl alcohol, polycaprolactone, polyvinylpyrrolidone; polysaccharides, such as cellulose; polypyrrole, etc.

[0103] Additionally, the hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticles or nanoparticle aggregates can be an agent that exhibits a specific group (R-) that can interact with the surface of the nanoparticles or nanoparticle aggregates, where R is typically selected from thiols, silanes, carboxylic acid groups, and phosphate groups.

[0104] When the core of the nanoparticle or nanoparticle aggregate is a conducting or semiconducting metal nanoparticle, R is preferably a thiol, thioether, thioester, dithiolane or carboxylic acid group. Preferably, the hydrophilic neutral coating agent is selected from thioglucose, 2-mercaptoethanol, 1-thioglycerol, thiodiglycol and hydroxybutyric acid.

[0105] When the core of the nanoparticle or nanoparticle aggregate is an insulator and oxide or mixed oxide nanoparticle, R is preferably a silane or phosphate group. Preferably, the hydrophilic neutral coating agent is hydroxymethyltriethoxysilane, fructose 6-phosphate or glucose 6-phosphate compound.

[0106] The hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticles or nanoparticle aggregates can be a zwitterionic compound, such as an amino acid, a peptide, a polypeptide, a vitamin, or a phospholipid.

[0107] The surface charge of nanoparticles or nanoparticle aggregates is typically determined by zeta potential measurements in water (aqueous solution) with a material concentration of nanoparticles or nanoparticle aggregates of 0.01 to 10 g / L, a pH of 6 to 8, and an electrolyte concentration (in water) of typically 0.001 to 0.2 M, e.g., 0.01 M or 0.15 M, as is well known to those skilled in the art. Under the conditions defined herein above, the surface charge of nanoparticles or nanoparticle aggregates is typically comprised between -10 mV and +10 mV (corresponding to a neutral surface charge), -20 mV to +20 mV, or -35 mV to +35 mV. When neutral, the surface charge of the nanoparticles or nanoparticle aggregates is typically comprised between -10 mV, -9 mV, -8 mV, -7 mV, -6 mV, -5 mV, -4 mV, -3 mV, -2 mV or -1 mV to 1 mV, 2 mV, 3 mV, 4 mV, 5 mV, 6 mV, 7 mV, 8 mV, 9 mV or 10 mV. If negative, the surface charge of the nanoparticles or nanoparticle aggregates is typically less than −11 mV, −12 mV, −13 mV, −14 mV, −15 mV, −16 mV, −17 mV, −18 mV, −19 mV, −20 mV, −21 mV, −22 mV, −23 mV, −24 mV, −25 mV, −26 mV, −27 mV, −28 mV, −29 mV, −30 mV, −31 mV, −32 mV, −33 mV, −34 mV, or −35 mV.

[0108] A complete biocompatible coating of the nanoparticles or aggregates can be advantageous in the context of the present invention to avoid any charge on the surface of the nanoparticles if the nanoparticles exhibit a hydrophilic neutral surface charge. By "complete coating" is meant the presence of such a high density / compactness of the biocompatible molecules that they are able to form at least a complete monolayer on the surface of the particle.

[0109] The biocompatible coating allows for the stabilization of the nanoparticles, in particular in fluids such as physiological fluids (blood, plasma, serum, etc.) or any isotonic or physiological medium required for pharmaceutical administration.

[0110] Stability can be confirmed by dry extract quantification using a drying oven, typically measured on nanoparticle suspensions before and after filtration through a 0.45 μm filter.

[0111] The coating has the advantage of preserving the integrity of the particle in vivo, ensuring or improving its biocompatibility, and facilitating its optional functionalization (e.g., with spacer molecules, biocompatible polymers, targeting agents, proteins, etc.).

[0112] The biocompatible nanoparticles or nanoparticle aggregates of the present invention should not release toxic species or exhibit redox behavior after in vivo administration (i.e., administration at physiological pH), typically in order for the nanoparticles or nanoparticle aggregates to be considered biocompatible, i.e., safe for use in subjects, particularly mammals, preferably humans.

[0113] Another particular object described herein relates to a composition, in particular a pharmaceutical composition, comprising the nanoparticles and / or nanoparticle aggregates as defined above, preferably together with a further pharmaceutically acceptable carrier or vehicle.

[0114] In particular, described herein are compositions for use in preventing or treating a neurological disorder or at least one symptom thereof as described herein in a subject without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing the nanoparticles or nanoparticle aggregates to any other external activation source, such as a light source, a magnetic field, or an ultrasound source, wherein the composition comprises or consists of nanoparticles and / or nanoparticle aggregates and a pharmaceutically acceptable support, wherein the nanoparticle or nanoparticle aggregate material is typically a conductive material, a semiconducting material, a material with a dielectric constant ε of 200 or greater, as described herein above, ijk Insulating materials with a dielectric constant ε of 100 or less ijk The insulating material is selected from the group consisting of:

[0115] In a preferred embodiment, the composition comprises or consists of at least two different nanoparticles and / or nanoparticle aggregates, each nanoparticle or nanoparticle aggregate consisting of a different material, typically a conductive material, a semiconductive material, a dielectric constant ε of 200 or greater, ijk Insulating materials with a dielectric constant ε of 100 or less ijk The insulating material is selected from the group consisting of:

[0116] In typical aspects of the present invention, the nanoparticles or nanoparticle aggregates described herein are not used as carriers of (active) therapeutic compounds or drugs.

[0117] In certain embodiments, the composition comprises the nanoparticles or nanoparticle aggregates of the present invention and can further comprise a therapeutic agent. In the context of the present invention, such a therapeutic agent is typically not a nanoparticle or nanoparticle aggregate. The therapeutic agent can be selected from any drug used to treat neurological disorders. The therapeutic agent is typically selected from antipsychotic agents, antidopaminergic agents, dopaminergic agents, anticholinergic agents, cholinergic agents, antiglutamatergic agents, glutamatergic agents, acetylcholinesterase inhibitors, N-methyl D-aspartate (NMDA) receptor antagonists, gamma-aminobutyric acid (GABA) agonists, botulinum toxin, antidystonic agents, antiepileptic agents, anticonvulsants, mood stabilizers, antidepressants, and sedatives.

[0118] The composition can be in the form of a solid, liquid (particles in suspension), aerosol, gel, paste, etc. Preferred compositions are in the form of a liquid or gel. Particularly preferred compositions are in the form of a liquid.

[0119] The pharmaceutically acceptable support or carrier employed can be any of the classic supports known to those skilled in the art, such as physiological saline, isotonic sterile buffer solutions, non-aqueous media solutions, and the like.

[0120] The composition may also include stabilizers, sweeteners, surfactants, polymers, and the like.

[0121] The compositions may be formulated, for example, as ampoules, aerosols, bottles, tablets or capsules using techniques of pharmaceutical formulation known to those skilled in the art.

[0122] The nanoparticles or nanoparticle aggregates of the present invention can be administered to a subject using a variety of possible routes, such as intracranial, intravenous (IV), airway (inhalation), intrathecal, intraocular or oral routes (oral), intracerebroventricular (ICV), preferably using intracranial or intrathecal routes.

[0123] Repeated injections or administrations of nanoparticles can be administered as needed. Preferably, the nanoparticles or nanoparticle aggregates should be administered in one dose.

[0124] Upon administration, the nanoparticles and / or nanoparticle aggregates typically interact with the neuronal subject. In preferred embodiments, this interaction is prolonged, i.e., over hours, days, weeks, or months. In certain embodiments, the nanoparticles or nanoparticle aggregates remain within the subject.

[0125] The nanoparticles or nanoparticle aggregates described herein, and compositions comprising such nanoparticles or nanoparticle aggregates, are for use in subjects, typically in animals, preferably in mammals, and even more preferably in humans, typically in human patients, regardless of their age or sex.

[0126] A typical amount of nanoparticles or nanoparticle aggregates administered to the cerebral cortex, hippocampus, and / or amygdala of a subject is 10 5 ~10 17 pieces, 10 5 ~10 16 Pieces or 10 5 ~10 15 pieces, preferably 10 7 ~10 14 10, more preferably 10 9 ~10 12 A typical amount of nanoparticles or nanoparticle aggregates administered to the cerebral cortex, hippocampus, and / or amygdala of a subject is 1 cm 3 per 10 nanoparticles or nanoparticle aggregates 2 ~10 12 There are individuals.

[0127] A typical amount of nanoparticles or nanoparticle aggregates administered to the deep brain of a subject is 10 4 ~10 17 pieces, 10 4 ~10 16 pieces, 10 4 ~10 15 Pieces or 10 4~10 14 pieces, preferably 10 6 ~10 12 10, more preferably 10 8 ~10 11 A typical amount of nanoparticles or nanoparticle aggregates administered deep into the brain of a subject is 1 cm 3 per 10 nanoparticles or nanoparticle aggregates 1 ~10 11 There are individuals.

[0128] Also described herein is a method for preventing or treating a neurological disorder or at least one symptom thereof in a subject, comprising administering to the subject any one of the nanoparticles or nanoparticle aggregates described herein. The method typically does not include any step of exposing the subject, or more precisely the nanoparticles or nanoparticle aggregates administered to the subject, to an electric field, and preferably does not include any step of exposing the subject, or more precisely the nanoparticles or nanoparticle aggregates administered to the subject, to any other external activation source, such as a light source, a magnetic field, or an ultrasound source.

[0129] A further object of the present invention is to provide a method for producing nanoparticles comprising or consisting of at least two different nanoparticles and / or at least two different nanoparticle aggregates as described herein, wherein each nanoparticle or nanoparticle aggregate of a different material is typically a conductive material, a semiconducting material, a dielectric constant ε of 200 or greater, as described herein. ijk Insulating materials with a dielectric constant ε of 100 or less ijk The kit relates to an insulating material having a thickness of 1000 nm to 1000 nm.

[0130] In certain embodiments, the kit contains, in different containers, different nanoparticles and / or nanoparticle aggregates described herein that are intended to be contacted, typically either in situ, i.e., mixed on the target site; or mixed in vitro or ex vivo prior to deposition of the mixture at the target site.

[0131] A further object relates to kits further comprising at least one additional therapeutic agent different from the nanoparticles or nanoparticle aggregates described herein. The therapeutic agent may be, for example, an antipsychotic, an antidopaminergic, a dopaminergic, an anticholinergic, a cholinergic, an antiglutamatergic, a glutamatergic, an acetylcholinesterase inhibitor, an N-methyl D-aspartate (NMDA) receptor antagonist, a gamma-aminobutyric acid (GABA) agonist, a botulinum toxin, an antidystonic agent, an antiepileptic, an anticonvulsant, a mood stabilizer, an antidepressant, or a sedative, and the selection of the therapeutic agent may be made by a person skilled in the art depending on the nature of the target disease. As explained herein above, such additional therapeutic agent is typically not a nanoparticle or nanoparticle aggregate.

[0132] Also described herein is the in vivo, in vitro, or ex vivo use of such kits in methods for preventing or treating a neurological disorder described herein, or at least one symptom thereof, in a subject, without exposing the nanoparticles or nanoparticle aggregates administered to the subject to an electric field, and preferably without exposing them to any other external activation source, such as a light source, magnetic field, or ultrasound source. Also disclosed herein is a kit described herein for use in preventing or treating a neurological disorder, or at least one symptom thereof, in a subject, without exposing the nanoparticles or nanoparticle aggregates administered to the subject to an electric field, and preferably without exposing them to any other external activation source, such as a light source, magnetic field, or ultrasound source.

[0133] At the neuronal level, nanoparticles have been described to enhance or inhibit the electrical excitability of neurons. For example, zinc oxide, carbon nanotubes, and gold nanoparticles have been found to enhance the electrical excitability of neurons, while copper oxide, silver, carbon black, iron oxide, and titanium oxide have been found to inhibit the electrical excitability of neurons (Polak P & Shefi O. Nanomedicine: Nanotechnology, Biology and Medicine 11 (2015) 1467-1479, Nanometric agents in the service of neuroscience: Manipulation of neuronal growth and activity using nanoparticles).

[0134] Studies on the systemic effects of coated silver nanoparticles (cAgNPs)—using the amphiphilic polymer polyethylene glycol—[cAgNPs with a hydrodynamic diameter of 13 nm ± 2 nm in pure water (dynamic light scattering technique) and a zeta potential of -69 mV (Zetasizer Nano)] on neuronal systems have shown that the nanoparticles induce changes in mechanisms affecting excitability. Furthermore, neuronal network simulations showed that localized cAgNP-induced changes led to changes in network activity throughout the network, indicating that local application of cAgNPs can affect the activity of the entire network (Busse M. et al. International Journal of Nanomedicine 2013:8 3559-3572, Estimating the modulatory effects of nanoparticles on neuronal circuits using computational upscaling).

[0135] It has also been described that the increased neuronal excitability associated with intracellular gold nanoparticles potentially has detrimental effects on neurons under pathological conditions, e.g., seizures (Jung S, et al. PLOS ONE 2014, 9(3) e91360, Intracellular gold nanoparticles increase neuronal excitability and aggravate seizure activity in the mouse brain).

[0136] The nanoparticles or nanoparticle aggregates of the present invention are for use in / to prevent or treat a neurological disease or at least one symptom thereof by normalizing the synchronization of oscillations within and / or between neuronal networks in different regions and / or between regions of the brain, without exposing said nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source.

[0137] As illustrated in Figures 2 and 3, communication within and / or between different brain regions is affected in neurological disorders. Depending on the neurological disorder and associated symptoms, exposure of specific brain regions to the nanoparticles of the present invention may improve communication by normalizing the synchronization of oscillations (i.e., normalizing coherence) within and / or between neuronal networks within and / or between different brain regions (Figures 4 and 5).

[0138] The following examples and their corresponding figures illustrate the invention without limiting its scope. [Brief explanation of the drawings]

[0139] [Figure 1] Figure 1. Schematic diagram of the brain (sagittal plane). [Figure 2] Figure 2. Hypersynchronization and desynchronization between two neuronal networks. [Figure 3]Figure 3. Brain regions involved in various neurological disorders. [Figure 4] Figure 4. Effect of nanoparticles (NPs) on normalizing hypersynchrony (movement disorder). [Figure 5] Figure 5. Effect of nanoparticles (NPs) on normalizing synchronization disorders (psychiatric and cognitive disorders). [Figure 6] Figure 6. Experimental scheme for Parkinson's disease induction with MPP+ treatment and electrical activity recording. Mouse ventral midbrain / cortex cocultures were prepared from E14.5 NMRI mice and cultured in 48-well MEAs for 3 weeks (total culture period). After 7 days of culture (day 7), the cultures were treated with nanoparticle suspension ("nanoparticle" group), GDNF (20 ng / ml) ("reference" group), or water ("control" group and "MPP+" group). On day 8, they were treated with MPP+ (20 μM) ("nanoparticle" group, "reference" group, and "MPP+" group) or water ("control" group). Spontaneous activity was recorded on day 21. [Figure 7] Figure 7. Two simplified burst schemes outlining some parameters that can be extracted from electrical activity recordings. Shown are parameters describing overall activity (spikes, bursts, interburst intervals (IBIs), and burst period) and burst structure (burst duration, burst plateau, burst amplitude, interburst spike interval (ISI), and burst area). The standard deviations (SD) of these parameters are measures of the regularity of overall activity and burst structure, respectively. The coefficient of variation in time (CVtime) reflects the temporal regularity of each unit's activity pattern. CVtime is calculated by the ratio of the parameter's standard deviation to its mean. The coefficient of variation across networks (CVnet) reflects the synchronization between neurons within the network. CVnet is calculated by the ratio of the parameter's standard deviation by its mean across the network. A larger CVnet value indicates a wider variation in activity across the network, implying less synchronization. [Figure 8-1]Figure 8-1. Functional effects observed in the "nanoparticle" group (nanoparticles from Example 1) and "reference" group compared to the "control" and "MPP+" groups on midbrain / cortex network activity. The data show MPP+-induced functional effects and demonstrate the preventative / rescue effects possible with the nanoparticles of the invention or GDNF (i.e., the ability to prevent / rescue functional effects to a level similar to that of the "control" group). [Figure 8-2] Figure 8-2. Functional effects observed in the "nanoparticle" group (nanoparticles from Example 2) and "reference" group compared to the "control" and "MPP+" groups on midbrain / cortex network activity. The data show MPP+-induced functional effects and demonstrate the preventative / rescue effects possible with the nanoparticles of the invention or GDNF (i.e., the ability to prevent / rescue functional effects to a level similar to that of the "control" group). [Figure 9] Figure 9. Functional effects observed in the "nanoparticle" group (nanoparticles from Examples 5 and 6) compared to the "control" and "MPP+" groups on midbrain / cortex network activity. The data show MPP+-induced functional effects and demonstrate the preventative / rescue effects possible with the nanoparticles of the invention (i.e., the ability to prevent / rescue functional effects to a level similar to that of the "control" group). [Figure 10] Figure 10. Effect score analysis for the "nanoparticle" group, "reference" group, "control" group and "MPP+" group. [Figure 11] Figure 11. Experimental scheme for induction, treatment and electrical activity recording of Alzheimer's disease with amyloid beta 1-42 (Abeta 1-42). After 4 weeks of culture (culture period), Abeta 1-42 (100 nM) ("nanoparticle" group, "reference" group and "Abeta" group) or water ("control" group) (T0) was added to the neuronal network. After 4 hours, nanoparticle suspension ("nanoparticle" group), donepezil (300 nM) ("reference" group) or water ("control" group and "Abeta" group) was added. Spontaneous activity was recorded as follows: -T0 (before addition of Abeta 1-42), -T0+1 h, +2 h, +3 h, +4 h (before addition of nanoparticles, donepezil or water), +5 h and +6 h. [Figure 12-1]Figure 12-1. Functional effects observed in the "nanoparticle" and "reference" groups compared to the "control" and "Abeta 1-42" groups on cortical network activity. The data show the Abeta 1-42 functional effects and demonstrate the rescue effect possible with the nanoparticles of the present invention (Example 1) or donepezil (i.e., the ability to rescue the functional effect to a level similar to that of the "control" group). [Figure 12-2] Figure 12-2. Functional effects observed in the "nanoparticle" and "reference" groups compared to the "control" and "Abeta 1-42" groups on cortical network activity. The data show the Abeta 1-42 functional effects and demonstrate the rescue effect possible with the nanoparticles of the present invention (Examples 2 and 3) or donepezil (i.e., the ability to rescue the functional effect to a level similar to that of the "control" group). [Figure 12-3] Figure 12-3. Functional effects observed in the "nanoparticle" and "reference" groups compared to the "control" and "Abeta 1-42" groups on cortical network activity. The data show the Abeta 1-42 functional effects and demonstrate the rescue effect possible with the nanoparticles of the present invention (Examples 4 and 5) or donepezil (i.e., the ability to rescue the functional effect to a level similar to that of the "control" group). [Figure 12-4] Figure 12-4. Functional effects observed in the "nanoparticle" and "reference" groups compared to the "control" and "Abeta 1-42" groups on cortical network activity. The data show the Abeta 1-42 functional effects and demonstrate the rescue effect possible with the nanoparticles of the present invention (Example 6) or donepezil (i.e., the ability to rescue the functional effect to a level similar to that of the "control" group). [Figure 13] Figure 13. Effect score analysis for the "nanoparticle" group, "reference" group, "control" group (effect score = 0) and "Abeta" group (effect score = 1). [Figure 14] Figure 14. Representative TEM images of gold nanoparticles from Example 9. The median maximum core sizes of the nanoparticles in the population are equal to 108 nm (Gold-110), 83 nm (Gold-80), 45 nm (Gold-45), 34 nm (Gold-30), and 15 nm (Gold-15), respectively. [Figure 15] FIG. 15. Effect score analysis for the "nanoparticle" group (gold-45 and gold-15 nanoparticles from Example 9), the "control" group (effect score=0) and the "MPP+" group (effect score=1). [Figure 16] Figure 16. Representative scanning electron microscope (SEM) image of PEDOT nanoparticles from Example 11. [Figure 17] FIG. 17. Effect score analysis for the "nanoparticle" group (PEDOT nanoparticles from Example 11), the "control" group (effect score=0) and the "MPP+" group (effect score=1).

[0140] Example In vitro studies of neurons At the neuronal level, the patch clamp technique is very useful for detecting action potentials because it allows simultaneous direct measurement and control of the neuronal membrane potential.

[0141] This technique is used to assess the effects of nanoparticles on single neurons.

[0142] In vitro studies of neuronal networks Isolated neuronal cultures coupled to multielectrode arrays (MEAs) have been widely used to better understand the complexity of brain networks. In addition, the use of isolated neuronal ensembles allows for the manipulation and control of network connectivity. MEA systems enable noninvasive, long-lasting, simultaneous extracellular recordings from multiple sites in neuronal networks in real time, improving spatial resolution and providing robust measurements of network activity. The simultaneous collection of action potential and field potential data over long periods of time allows for the monitoring of network function resulting from the interplay of all cellular mechanisms responsible for spatiotemporal pattern generation (Johnstone AFM et al., Neurotoxicology (2010), 31: 331-350, Microelectrode arrays: a physiologically based neurotoxicity testing platform for the 21 stcentury). Compared with patch clamp and other single-electrode recording techniques, MEA measures responses across the entire network and integrates global information about the interactions of all receptors, synapses, and neuronal types present in the network (Novellino A. et al., Frontiers in Neuroengineering. (2011), 4(4), 1-14, Development of microelectrode array-based tests for neurotoxicity: assessment of interlaboratory reproducibility with neuroactive chemicals.). Therefore, MEA recordings have been used to understand neuronal communication, information encoding, propagation, and processing in neuronal cultures (Taketani, M and Baudry, M. (2006). Advances in Network Electrophysiology. New York, NY: Springer; Obien et al., Frontiers in Neurosciences, 2015, 8(423): Revealing neuronal functions through microelectrode array recordings). The MEA technology is an advanced phenotypic, high-content screening method for characterizing functional changes in network activity in cultures of electrically active cells, with great sensitivity for aspects of neurogenesis, as well as neuroregeneration and neurodegeneration.Furthermore, neuronal networks grown on MEAs are known to be responsive to neuroactive or neurotoxic compounds in concentrations similar to those that alter the function of the uninjured mammalian nervous system (Xia et al., Alcohol, 2003, 30, 167-174: Histiotypic electrophysiological responses of cultured neuronal networks to ethanol; Gramowski et al., European Journal of Neuroscience, 2006, 24, 455-465: Functional screening of traditional antidepressants with primary cortical neuronal networks grown on multielectrode neurochips; Gramowski et al., Frontiers in Neurology, 2015, 6(158): Enhancement of cortical network activity in vitro and promotion of GABAergic neurogenesis by stimulation with an electromagnetic field with a 150 MHz carrier wave pulsed with an alternating 10 and 16 Hz modulation).

[0143] This technique is used to evaluate the effects of nanoparticles on neuronal networks.

[0144] In vivo studies of neuronal networks Suitable animal models for assessing the effects of the nanoparticles of the present invention on the neuronal networks of animals are contemplated.

[0145] For example, a mouse model of Parkinson's disease is used to assess the effect of nanoparticles on reducing behavioral deficits (movement disorders), and a rat or mouse model of Alzheimer's disease is used to assess the effect of nanoparticles on spatial learning and memory impairment (cognitive impairment) in animals.

[0146] Example 1. Nanoparticles Prepared with Conductive Materials: Synthesis of Gold Nanoparticles Coated with a Biocompatible Coating with Neutral Surface Charge Gold nanoparticles were synthesized by reducing gold chloride (HAuCl4) with a capping agent (sodium citrate) (protocol adapted from G. Frens, Nature Physical Science 241 (1973) 21). In a typical experiment, the HAuCl4 solution was heated to boiling. Subsequently, sodium citrate solution was added. The resulting solution was kept boiling for an additional 5 minutes. The nanoparticle suspension was filtered through a 0.22 μm filter (filter membrane: poly(ether sulfone) (PES)), and the gold concentration in the suspension was determined by UV-visible spectroscopic assay at 530 nm.

[0147] Surface coating was performed using α-methoxy-ω-mercaptopoly(ethylene glycol) 20 kDa ("Thiol-PEG 20 kDa"). A sufficient amount of "Thiol-PEG 20 kDa" was added to the nanoparticle suspension to reach at least half a monolayer coverage on the gold nanoparticle surface (2.5 molecules / nm 2 The pH was adjusted to 7-7.2, and the nanoparticle suspension was stirred overnight.

[0148] The nanoparticle suspension was diluted in water (final concentration: [Au] = 0.1 g / L), and the hydrodynamic diameter (a measure of intensity) was determined at room temperature (approximately 25 °C) by dynamic light scattering (DLS) on a Nano-Zetasizer (Malvern) at a scattering angle of 173° with a laser emitting at 633 nm. The hydrodynamic diameter of the suspension of biocompatible gold nanoparticles thus obtained was found to be equal to 118 nm, and the polydispersity index (dispersion of the population of nanoparticles with respect to size) was found to be 0.13.

[0149] The zeta potential was determined by diluting the nanoparticle suspension in 1 mM NaCl solution at pH 7 (final concentration: [Au] = 0.1 g / L) and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern). The zeta potential at pH 7 was found to be equal to -1 mV.

[0150] Example 2. Nanoparticles prepared with conductive materials: Synthesis of gold nanoparticles coated with a biocompatible coating with a negative surface charge Gold nanoparticles were prepared as described in Example 1 (same gold inorganic core).

[0151] After 0.22 μm filtration through a PES membrane filter, the gold concentration in the suspension was determined by UV-visible spectrophotometric assay at 530 nm.

[0152] Biocompatible surface coating was achieved using meso-2,3-dimercaptosuccinic acid (DMSA). A sufficient amount of DMSA was added to the nanoparticle suspension to reach at least half a monolayer coverage on the surface (2.5 molecules / nm). 2 The pH was adjusted to 7-7.2, and the nanoparticle suspension was stirred overnight.

[0153] The nanoparticle suspension was diluted in water (final concentration: [Au] = 0.1 g / L), and the hydrodynamic diameter (a measure of intensity) was determined at room temperature (approximately 25 °C) by dynamic light scattering (DLS) on a Nano-Zetasizer (Malvern) at a scattering angle of 173 ° with a laser emitting at 633 nm. The hydrodynamic diameter of the nanoparticles in suspension thus obtained was equal to 76 nm, and the polydispersity index (dispersion of the population of nanoparticles with respect to size) was 0.46.

[0154] The zeta potential was determined by diluting the nanoparticle suspension in 1 mM NaCl solution at pH 7 (final concentration: [Au] = 0.1 g / L) and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern). The zeta potential at pH 7 was found to be equal to -23 mV.

[0155] Example 3. Nanoparticles prepared by insulating materials with low dielectric constants below 100: Synthesis of zirconium oxide nanoparticles coated with a biocompatible coating with neutral surface charge Zirconium oxide (ZrO2) nanoparticles were synthesized by precipitation of zirconium chloride (ZrCl4) with tetramethylammonium hydroxide (TMAOH) at basic pH. The resulting suspension was transferred to an autoclave and heated at temperatures above 110 °C. After cooling, the suspension was washed with deionized water and acidified.

[0156] The median maximum core size of the nanoparticles or nanoparticle aggregates in the population, as well as the size of the cores of nanoparticles or nanoparticle aggregates representing 30% to 70% of the population of nanoparticles and nanoparticle aggregates, were assessed using transmission electron microscopy and found to be equal to 10 nm and 8 to 12 nm, respectively. 446 nanoparticles were counted and their maximum dimensions measured.

[0157] After 0.22 μm filtration on a PES membrane filter, the (ZrO2) nanoparticle concentration was determined by drying the aqueous solution to a powder and weighing the resulting mass. Biocompatible coatings were prepared using silane-poly(ethylene)glycol 2 kDa ("Si-PEG 2 kDa"). A sufficient amount of "Si-PEG 2 kDa" was added to the nanoparticle suspension to achieve at least half a monolayer coverage on the surface (2.5 molecules / nm2). 2 The nanoparticle suspension was stirred overnight and subsequently the pH was adjusted to 7.

[0158] The nanoparticle suspension was diluted in water (final concentration of ZrO2 constituting the nanoparticle core: [ZrO2] = 0.1 g / L), and the hydrodynamic diameter (a measure of intensity) was determined at room temperature (approximately 25 °C) by dynamic light scattering (DLS) on a Nano-Zetasizer (Malvern) at a scattering angle of 173° with a laser emitting at 633 nm. The hydrodynamic diameter of the nanoparticles was found to be equal to 55 nm, and the polydispersity index (dispersion of the population of nanoparticles with respect to size) was found to be 0.1.

[0159] The zeta potential was determined by diluting the nanoparticle suspension in 1 mM NaCl solution at pH 7 (final concentration: [ZrO] = 0.1 g / L) and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern). The zeta potential at pH 7 was found to be equal to -1 mV.

[0160] Example 4. Nanoparticles prepared by insulating materials with low dielectric constants below 100: Synthesis of zirconium oxide nanoparticles coated with a biocompatible coating with negative surface charge Zirconium oxide nanoparticles were prepared as described in Example 3 (same inorganic core).

[0161] 0.22 μm filtration on a PES membrane filter was performed and the (ZrO 2 ) nanoparticle concentration was determined by drying the aqueous suspension to a powder and weighing the resulting mass.

[0162] Surface functionalization was performed using sodium hexametaphosphate. A sufficient amount of sodium hexametaphosphate was added to the nanoparticle suspension to reach at least half a monolayer coverage on the surface (2.5 molecules / nm 2 The nanoparticle suspension was stirred overnight and subsequently the pH was adjusted to 7.

[0163] The nanoparticle suspension was diluted in water (final concentration of ZrO2 constituting the nanoparticle core: [ZrO2] = 0.1 g / L), and the hydrodynamic diameter (a measure of intensity) was determined at room temperature (approximately 25 °C) by dynamic light scattering (DLS) on a Nano-Zetasizer (Malvern) at a scattering angle of 173° with a laser emitting at 633 nm. The hydrodynamic diameter of the nanoparticles was found to be equal to 70 nm, and the polydispersity index (dispersion of the population of nanoparticles with respect to size) was found to be 0.11.

[0164] The zeta potential was determined by diluting the nanoparticle suspension in 1 mM NaCl solution at pH 7 (final concentration: [ZrO] = 0.1 g / L) and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern). The zeta potential at pH 7 was found to be equal to -33 mV.

[0165] Example 5. Nanoparticles prepared with semiconductor materials: Silicon (Si) nanoparticles coated with a biocompatible coating having a negative surface charge

[0166] Silicon (Si) nanoparticles (powder) were obtained from US Research Nanomaterials Inc. They were coated with PVP (1 wt%), which had a density of 0.1 molecules / nm on the surface. 2 Indicates less than.

[0167] They were dispersed in water at 30 g / L under ultrasonication (by probe).

[0168] 0.22 μm filtration on a PES membrane filter was performed and the (Si) nanoparticle concentration was determined by drying the suspension to a powder and weighing the resulting mass.

[0169] The nanoparticle suspension was diluted in water (final concentration of Si constituting the nanoparticle core: [Si] = 0.1 g / L), and the hydrodynamic diameter (a measure of intensity) was determined at room temperature (approximately 25 °C) by dynamic light scattering (DLS) on a Nano-Zetasizer (Malvern) at a scattering angle of 173° with a laser emitting at 633 nm. The hydrodynamic diameter of the nanoparticles was found to be equal to 164 nm, and the polydispersity index (dispersion of the population of nanoparticles with respect to size) was found to be 0.16.

[0170] The median maximum core size of the nanoparticles or nanoparticle aggregates in the population, as well as the size of the cores of nanoparticles or nanoparticle aggregates representing 30% to 70% of the population of nanoparticles and nanoparticle aggregates, were assessed using transmission electron microscopy and found to be equal to 53 nm and 45 to 61 nm, respectively. 71 nanoparticles were counted and their maximum dimensions measured.

[0171] The zeta potential was determined by diluting the nanoparticle suspension in 1 mM NaCl solution at pH 7 (final concentration: [Si] = 0.1 g / L) and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern). The zeta potential at pH 7 was found to be equal to -19 mV.

[0172] Example 6. Nanoparticles prepared from insulating materials with high dielectric constants of 200 or more: Barium titanate nanoparticles coated with a biocompatible coating having a negative surface charge A suspension of barium titanate (BaTiO3) nanoparticles (20 wt % in water) was obtained from US Research Materials Inc. (US3835).

[0173] Surface functionalization was performed using silane-poly(ethylene)glycol 10 kDa ("Si-PEG 10 kDa"). Briefly, "Si-PEG 10 kDa" was first dissolved in an ethanol / water solution (1 / 3 v / v) and added to a BaTiO suspension (20 wt % in water) to achieve complete monolayer coverage on the nanoparticle surface. The suspension was sonicated and subsequently stirred overnight. After 0.22 μm filtration (filter membrane: poly(ether sulfone)), a washing step was performed to remove unreacted "Si-PEG 10 kDa" polymer.

[0174] The nanoparticle suspension was diluted in water (final concentration of BaTiO3 constituting the nanoparticle core: [BaTiO3] = 0.1 g / L), and the hydrodynamic diameter (a measure of intensity) was determined at room temperature (approximately 25 °C) by dynamic light scattering (DLS) on a Nano-Zetasizer (Malvern) at a scattering angle of 173° with a laser emitting at 633 nm. The hydrodynamic diameter of the nanoparticles was found to be equal to 164 nm, and the polydispersity index (dispersion of the population of nanoparticles with respect to size) was found to be 0.16.

[0175] The zeta potential was determined by diluting the nanoparticle suspension in 1 mM NaCl solution at pH 7 (final concentration: [BaTiO] = 0.1 g / L) and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern). The zeta potential at pH 7 was found to be -11 mV.

[0176] The median maximum core size of the nanoparticles or nanoparticle aggregates in the population, as well as the size of the cores of nanoparticles or nanoparticle aggregates representing 30% to 70% of the population of nanoparticles and nanoparticle aggregates, were assessed using transmission electron microscopy and found to be equal to 67 nm and 60 to 77 nm, respectively. 51 nanoparticles were counted and their maximum dimensions measured.

[0177] Example 7. MPP using phenotypic MEA screening technology +Evaluation of the preventive / rescue efficacy of nanoparticles from Examples 1, 2, 5 and 6 in evoked neuronal networks MPPs cultured on a 48-well MEA for 3 weeks + The preventive / rescue efficacy of the nanoparticles of the present invention was tested on ventral midbrain / cortex co-cultures of treated mice. This model represents an in vitro Parkinson's model for screening compounds based on the functional rescue of dopaminergic neurons using damaged midbrain / cortex cultures grown on MEA. The midbrain is a brain region that includes the substantia nigra, which is part of the basal ganglia and contains the majority of dopaminergic neurons. The preventive / rescue effect of the nanoparticles was evaluated by measuring the extracellular electrical activity of neuronal co-cultures seeded on microelectrode array (MEA) chips.

[0178] In vitro induction of parkinsonian phenotype in mouse neurons using 1-methyl-4-phenylpyridinium iodide (MPP) + There is strong evidence that mitochondrial dysfunction plays a role in the pathogenesis of Parkinson's disease (PD). + It has been found to be a mitochondrial poison that inhibits cellular respiration via blocking the electron transport enzyme complex I (NADH:ubiquinone oxidoreductase). Several laboratories have reported that there is a selective defect in complex I of the mitochondrial electron transport chain in the substantia nigra of postmortem tissue from PD patients, and that complex I activity is also reduced in platelets from early PD patients. Drugs, such as glial cell line neurotrophic factor (GDNF), act as neuroprotectants, and MPP +GDNF has shown promising preclinical results in preventing / relieving the effects of Parkinson's disease. GDNF is often used as a reference in experimental preclinical protocols (Peng J. et al., Journal of Biomolecular screening, 2013, 18(5), 522-533: Using human pluripotent stem cell-derived dopaminergic neurons to evaluate candidate Parkinson's disease therapeutic agents in MPP+ and rotenone models.).

[0179] Materials and Methods Primary cell culture, treatment conditions Midbrain and frontal cortex tissues were collected from embryonic day 14.5 chr:NMRI mice (Charles River). Mice were sacrificed by cervical dislocation. Tissues were dissociated by enzymatic digestion (133.3 Kunitz units / ml DNase; 10 units / ml papain) and mechanical trituration, counted, and controlled for vitality. They were seeded onto MEAs in 20 μl drops of DMEM containing laminin (10 μg / ml), 10% fetal bovine serum, and 10% horse serum. Cultures on MEAs were incubated at 37°C in a 10% CO atmosphere until ready for use. DMEM containing 10% horse serum was replenished twice weekly.

[0180] In the "nanoparticle" group, wells were treated on day 7 with nanoparticle suspensions from Examples 1 ([Au] = 800 μM), 2 ([Au] = 800 μM), 5 ([Si] = 800 μM) and Example 6 ([BaTiO] = 2000 μM), followed on day 8 by 20 μM MPP + In the "control" group, water was added to the wells on the 7th day, followed by water on the 8th day. + In the " group, water was added to the wells on the 7th day, followed by 20 μM MPP on the 8th day. +In the "reference" group, GDNF (20 ng / ml) was added to the wells on day 7, followed by 20 μM MPP on day 8. + Added.

[0181] MPP + (or water for the "control" group), 24 hours after addition, the medium was changed and MPP + Washout of the cells was achieved. Thereafter, the medium was changed twice a week. GDNF was added only to the "reference" group at each medium change.

[0182] On day 21, 120 min of neuronal activity was recorded and 30 min of stable activity was analyzed (Fig. 6).

[0183] Microelectrode Arrays and Neurochips 48-well microelectrode array neurochips were purchased from Axion Biosystems Inc. These chips contain 16 passive electrodes per well. The surfaces were coated with polyethyleneimine (PEI, 50% in borate buffer) for 1 hour, washed, and air-dried.

[0184] Multichannel recording, and multiparameter data analysis Recordings were performed using a multichannel MAESTRO recording system from Axion Biosystems (USA). For extracellular recordings, 48-well MEAs were placed in a MAESTRO recording station and maintained at 37°C. Recordings were performed in DMEM / 10% heat-inactivated horse serum. The pH was maintained at 7.4 with a continuous flow of filtered, humidified air containing 10% CO2.

[0185] Each unit represents the activity arising from one neuron recorded with one electrode. Units were isolated at the beginning of recording. For each unit, action potentials (i.e., spikes) were recorded as spike trains, a group of states called a "burst." Bursts were quantitatively described by direct spike train analysis using the programs Spike Wrangler and NPWaveX (both from NeuroProof GmbH, Rostock, Germany). Bursts were defined by the onset and end of short spike events (Figure 7).

[0186] Multiparameter high-content analysis of network activity patterns extracted 204 activity-descriptive spike train parameters that allow for a precise description of activity changes in four categories: overall activity, burst structure, oscillatory behavior, and synchrony. - Changes in "global activity parameters" describe the effects on action potential discharge rate (spike rate), burst rate and burst period as time between bursts. - "Burst structure parameters" define the internal structure of spikes within a high frequency spike period ("burst"), e.g., spike frequency within a burst, spike rate within a burst, and burst spike density, as well as the overall structure of the burst, e.g., duration, area, and plateau. - "Oscillatory parameters" quantify the regularity of the occurrence or structure of bursts, this regularity being calculated by the coefficient of variation of the main activity parameters, which describes the variability of the parameters (global activity, burst structure) within an experimental case (Gramowski A. et al., Eur. J. Neurosci., 2004, 19, 2815-2825: Substance identification by quantitative characterization of oscillator activity in murine spinal cord networks on microelectrode arrays). Higher values ​​indicate a less regular burst structure or a less regular global activity (e.g. spikes, bursts). As a measure of synchrony in spike trains, the "CVnet parameter" reflects the "synchronization" between neurons within a network (Gramowski A. et al., Eur. J. Neurosci., 2004, 19, 2815-2825: Substance identification by quantitative characterization of oscillator activity in murine spinal cord networks on microelectrode arrays). CVnet is the coefficient of variation across the network. A larger CVnet value indicates a wider variation in activity across the network and less synchronization (Gramowski A. et al., Frontiers in Neurology, 2015, 6(158): Enhancement of cortical network activity in vitro and promotion of GABAergic neurogenesis by stimulation with an electromagnetic field with a 150MHz carrier wave pulsed with an alternating 10 and 16 Hz modulation).

[0187] MPP + The functional effects induced on neuronal networks and the preventive / rescue efficacy of the nanoparticles of the invention were assessed through the above parameters (and some of them are summarized in Table 2 below).

[0188] [Table 2]

[0189] Values ​​related to spontaneous activity on day 21 were obtained from 60-second segments of data obtained from a 30-minute time span after 30 minutes of activity stabilization. Results (parameter values) were expressed as the mean ± SEM of independent networks. For each "nanoparticle" group, at least 8 active wells were used, for the "control" group, at least 30 active wells were used, and for the "MPP" group, at least 10 active wells were used. + For the "group," at least 26 active wells ("active" means wells with a sufficient number of electrodes to measure electrical activity) were included in the analysis. Absolute parameter distributions were tested for normality, and statistically significant differences between groups were assessed by one-way ANOVA.

[0190] 8 and 9 show some representative parameters from the following categories: overall activity, oscillatory behavior, and synchrony. These parameters allow us to estimate MPP + The induced functional effect and the preventative / remedial efficacy of the nanoparticles of the invention or GDNF (ie, the ability to prevent / remediate the functional effect to a level similar to that of the "control" group) are characterized.

[0191] To assess compound efficacy, the results of the multiparameter selection of 204 parameters were expressed as a single parameter called the "Effect Score." This is a linear combination of the selected features, dividing the dataset into a "control" group with a mean value of "0" and a "MPP" group with a mean value of "1." + The calculation of the Z factor of the effect score was performed by selecting 18 features out of 204 parameters, and dividing the group into a "control" group and a "MPP" group. +The z' factor was optimized to find the best discrimination between the 'groups' (Kuemmel A, et al. J Biomol Screen., 2010, 15(1), 95-101: Integration of multiple readouts into the z' factor for assay quality assessment).

[0192] The effect score analysis is shown in Figure 10.

[0193] The preventive / remedial efficacy of the nanoparticles of the present invention is shown in Table 3.

[0194] [Table 3]

[0195] 8, 9, and 10 and Table 3 show that pre-processing of the neuronal network with the nanoparticles of the present invention significantly improved the MPP in the neuronal network. + It shows that the induced functional effects are prevented / relieved. Interestingly, the preventive / relief efficacy is observed for the category parameters related to oscillatory behavior and synchrony, and can reach the level observed in the "control" group. These oscillatory behavior and synchronization parameters are typically monitored as a measure of altered network development. Advantageously, these parameters can be rescued in the presence of the nanoparticles of the present invention.

[0196] These results suggest that MPP in neuronal networks + This highlights the advantageous ability of the nanoparticles described herein to prevent / relieve induced functional effects.

[0197] Example 8: Evaluation of the effect of nanoparticles from Examples 1, 2, 3, 4, 5 and 6 on amyloid beta 1-42 induced functional effects in primary mouse neuronal networks using phenotypic MEA screening technology The rescue efficacy of the nanoparticles of the present invention was tested in vitro via MEAs in an amyloid beta 1-42 (Abeta 1-42)-induced model of Alzheimer's disease in frontal cortex cultures of mouse neurons.

[0198] β-Amyloid peptide 1-42, a major component of neuritic plaques in patients with Alzheimer's disease (AD), induces excessive glutamate in the synaptic cleft by inhibiting astroglial glutamate transporters, and increases intracellular Ca via enhanced N-methyl-D-aspartate (NMDA) receptor activity. 2+ It is known that aβ increases levels of glutamatergic NMDA receptors. Other mechanisms that cause excitotoxicity may include the induction of oxidative stress and the direct effect of aβ on glutamatergic NMDA receptors. Regardless of the precise underlying pathogenic process, excessive stimulation of neurons by glutamate and intracellular calcium accumulation ultimately leads to neuronal apoptosis and disruption of synaptic plasticity. This dysregulation results in significant impairment of learning and memory function (Nyakas C. et al., Behavioral Brain Research, 2011, 221, 594-603: The basal forebrain cholinergic system in aging and dementia. Rescuing cholinergic neurons from neurotoxic amyloid-β42 with memantine.). Currently, FDA-approved anti-AD drugs are limited to acetylcholinesterase (AChE) inhibitors and NMDA receptor antagonists. Traditional AChE inhibitors include donepezil, which primarily acts on the central site of action of AChE.

[0199] Materials and Methods primary cell culture Frontal cortical tissue was collected from embryonic day 15 / 16 chr:NMRI mice (Charles River). Mice were sacrificed by cervical dislocation. Tissue was dissociated by enzymatic digestion (133.3 Kunitz units / ml DNase; 10 units / ml papain) and mechanical trituration, counted, and viability controlled. The tissue was seeded onto MEAs in 20 μl drops of DMEM containing laminin (10 μg / ml), 10% fetal bovine serum, and 10% horse serum. Cultures on MEAs were incubated at 37°C in a 10% CO atmosphere until ready for use. DMEM containing 10% horse serum was replenished to the culture medium twice weekly. Developing co-cultures were treated with the mitotic inhibitors 5-fluoro-2'-deoxyuridine (25 μM) and uridine (63 μM) 5 days after seeding to prevent further glial cell proliferation.

[0200] To induce Alzheimer's-related functional phenotypes, we used a subtoxic dose (100 nM) of synthetic HFIP (hexafluoroisopropanol)-treated Abeta 1-42 peptide (HFIP treatment generates amyloid beta monomers).

[0201] In the "nanoparticle" group, wells were first treated with Abeta 1-42 (synthetic HFIP-treated amyloid-beta 1-42 peptide) at time T0 (T0 being the end of the 28-day in vitro culture period). Then, in independent and parallel experiments, wells were treated with nanoparticle suspensions from Examples 1 ([Au] = 800 μM), 2 ([Au] = 800 μM), 3 ([ZrO2] = 800 μM), 4 ([ZrO2] = 800 μM), 5 ([Si] = 800 μM), and 6 ([BaTiO3] = 2000 μM) at time T0 + 4. In the "control" group, water was added to the wells at time T0 and then at time T0 + 4. In the "Abeta" group, Abeta 1-42 was added to the wells at time T0, and then water was added to the wells at time T0 + 4. In the "reference" group, Abeta 1-42 was added to the wells at T0 and donepezil (300 nM) was added to the wells at T0+4 hours.

[0202] Neuronal activity was recorded as follows (see Fig. 11). - Time TO, before addition of Abeta 1-42 (or water in the "control" group). -T0+1h, T0+2h, T0+3h, T0+4h (before adding nanoparticles in the nanoparticle group; before adding donepezil in the reference group; before adding water in the control group), T0+5h and T0+6h.

[0203] After 30 minutes of activity stabilization, values ​​were obtained from 60-second segments of data taken from a 30-minute time span.

[0204] Microelectrode Arrays and Neurochips 48-well microelectrode array neurochips were purchased from Axion Biosystems Inc. These chips contain 16 passive electrodes per well. The surfaces were coated with polyethyleneimine (PEI, 50% in borate buffer) for 1 hour, washed, and air-dried.

[0205] Multichannel recording and multiparameter data analysis A multichannel MAESTRO recording system from Axion Biosystems (USA) was used for recording. For extracellular recordings, 48-well MEAs were placed in a MAESTRO recording station and maintained at 37°C. Recordings were performed in DMEM / 10% heat-inactivated horse serum. The pH was maintained at 7.4 with a continuous flow of filtered, humidified air containing 10% CO2. Action potentials, or "spikes," were recorded in spike trains, which represent a group of states known as "bursts." Bursts were quantitatively described by direct spike train analysis using the programs Spike Wrangler and NPWaveX (both NeuroProof GmbH, Rostock, Germany). Bursts were defined by the onset and end of short spike events.

[0206] Multiparameter high-content analysis of network activity patterns extracted 204 activity-descriptive spike train parameters that allow for a precise description of activity changes in four categories: overall activity, burst structure, oscillatory behavior, and synchrony.

[0207] The functional effects of amyloid beta 1-42 on neuronal networks and the efficacy of rescuing the functional effects of neuronal networks by the nanoparticles of the invention were assessed through the above parameters (and some of them are summarized in Table 4 below).

[0208] [Table 4]

[0209] After 30 minutes of activity stabilization, values ​​related to spontaneous spontaneous activity were obtained from 60-second bins of data obtained from a 30-minute time span. Results (parameter values) were expressed as the mean ± SEM of independent networks. For each "nanoparticle" group, at least 9 active wells were included in the analysis; for the "control" group, at least 18 active wells; for the "Abeta" group, at least 18 active wells ("active" refers to wells with a sufficient number of electrodes to measure electrical activity). Absolute parameter distributions were tested for normality, and statistical significance between groups was assessed by one-way ANOVA.

[0210] Figure 12 shows some representative parameters from the following categories: overall activity, burst structure, oscillatory behavior and synchrony, which characterize the Abeta 1-42 functional effect and rescue efficacy (i.e., the ability to rescue the functional effect to a level similar to that of the "control" group) enabled by the nanoparticles of the present invention or donepezil.

[0211] To assess compound efficacy, the multiparameter results of selecting 204 parameters were expressed as a single parameter called the "effect score." This is a linear combination of the selected features, converting the dataset into a vector with a "control" group with a mean value of "0" and an "Abeta" group with a mean value of "1." The Z factor of the effect score was calculated by selecting 15 features from the 204 parameters and optimizing it to find the best discrimination between the "control" and "Abeta" groups (Kuemmel A, et al. J Biomol Screen., 2010, 15(1), 95-10: Integration of multiple readouts into the z' factor for assay quality assessment).

[0212] The effect score analysis is shown in Figure 13.

[0213] The rescue efficacy of the nanoparticles of the present invention is shown in Table 5.

[0214] [Table 5]

[0215] Figures 12 and 13 and Table 5 show that treatment of neuronal networks with nanoparticles of the present invention rescues the Abeta 1-42-induced functional effects of neuronal networks. Rescue efficacy is observed for categories of parameters related to oscillatory behavior and synchrony, which can advantageously reach the levels observed in the "control" group. These oscillatory behavior and synchronization parameters are classically evaluated to detect altered network development. Oscillatory behavior and synchronization can be rescued in the presence of nanoparticles of the present invention.

[0216] These results highlight the advantageous performance of the nanoparticles described in this application in rescuing Abeta 1-42-induced functional effects in neuronal networks.

[0217] Example 9: Synthesis and physicochemical characterization of gold nanoparticles of different sizes with neutral surface charge Gold nanoparticles are obtained by reduction of gold chloride with sodium citrate in aqueous solution. The protocol was adapted from G. Frens Nature Physical Science 241 (1973) 21.

[0218] In a typical experiment, the HAuCl solution is heated to boiling. Subsequently, the sodium citrate solution is added. The resulting suspension is kept boiling for an additional 5 minutes.

[0219] The nanoparticle size is tuned from about 15 nm to about 110 nm by carefully changing the ratio of citrate to gold precursor (see Table 6).

[0220] The as-prepared gold nanoparticle suspension is then concentrated using an ultrafiltration device (Amicon stirred cell model 8400 from Millipore) equipped with a cellulose membrane with appropriate molecular weight exclusion (MWCO) and filtered through a 0.22 μm exclusion membrane filter (PES membrane from Millipore) under a laminar flow hood.

[0221] Surface coating is performed using α-methoxy-ω-mercaptopoly(ethylene glycol) 20 kDa ("Thiol-PEG 20 kDa"). A sufficient amount of "Thiol-PEG 20 kDa" is added to the nanoparticle suspension to obtain a monolayer coating on the gold nanoparticle surface. The pH is adjusted to 6.8-7.4, and the nanoparticle suspension is stirred overnight. Excess Thiol-PEG 20 kDa is removed under a laminar flow hood using an ultrafiltration centrifugal filter with an appropriate MWCO membrane (Vivaspin from Sartorius or Amicon Ultra from Merck Millipore), and the final suspension is stored at 4 °C.

[0222] Particle size was measured using a transmission electron microscope by counting at least 200 nanoparticles and taking the largest nanoparticle dimension for size measurement. The median maximum core size of the nanoparticles or nanoparticle aggregates in the population, as well as the core sizes of nanoparticles or nanoparticle aggregates representing 30% to 70% of the population of nanoparticles and nanoparticle aggregates, are reported in Table 6. The gold ([Au]) concentration, measured by inductively coupled plasma optical emission spectroscopy (ICP-OES), and the zeta potential, determined by diluting a suspension of nanoparticles in 1 mM NaCl solution to a gold concentration ([Au]) of 0.01 to 0.05 g / L and a pH of approximately 7, and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern), are also reported in Table 6.

[0223] [Table 6]

[0224] FIG. 14 shows representative transmission electron microscope (TEM) images of the gold nanoparticles listed in Table 6.

[0225] Example 10. MPPs using phenotypic MEA screening technology + Evaluation of the preventive / rescue efficacy of nanoparticles gold-15 and gold-45 from Example 9 on induced neuronal networks The preventive / rescue efficacy of the nanoparticles of the present invention was evaluated by measuring the number of MPPs cultured in a 48-well MEA for 3 weeks. + The preventive / rescue effects of nanoparticles were evaluated in treated mouse ventral midbrain / cortex co-cultures by measuring the extracellular electrical activity of neuronal co-cultures seeded on microelectrode array (MEA) chips.

[0226] In vitro induction of the parkinsonian phenotype in mouse neurons was achieved using 1-methyl-4-phenylpyridinium iodide (MPP) + ) was carried out.

[0227] Materials and Methods Primary cell culture, treatment conditions Midbrain and frontal cortex tissues were collected from embryonic day 14.5 chr:NMRI mice (Charles River). Mice were sacrificed by cervical dislocation. Tissues were dissociated by enzymatic digestion (133.3 Kunitz units / ml DNase; 10 units / ml papain) and mechanical trituration, counted, and viability controlled. They were then seeded onto MEAs in 20 μl drops of DMEM containing laminin (10 μg / ml), 10% fetal bovine serum, and 10% horse serum. Cultures on MEAs were incubated at 37°C in a 10% CO atmosphere until ready for use. DMEM containing 10% horse serum was replenished twice weekly.

[0228] In the "nanoparticle" group, wells were treated with nanoparticle suspension ([Au]=310+ / -40 μM) from Example 9 (gold-15 and gold-45) on day 7, followed by 20 μM MPP on day 8. + In the "control" group, water was added to the wells on the 7th day, followed by water on the 8th day. + In the " group, water was added to the wells on the 7th day, followed by 20 μM MPP on the 8th day. + Added.

[0229] MPP + (or water for the "control" group), 24 hours after addition, the medium was changed and MPP + After that, the medium was changed twice a week.

[0230] On day 21, 120 min of neuronal activity was recorded and 30 min of stable activity was analyzed.

[0231] Microelectrode Arrays and Neurochips 48-well microelectrode array neurochips were purchased from Axion Biosystems Inc. These chips contain 16 passive electrodes per well. The surfaces were coated with polyethyleneimine (PEI, 50% in borate buffer) for 1 hour, washed, and air-dried.

[0232] Multichannel recording and multiparameter data analysis A multichannel recording system from Axion Biosystems (USA) was used for recording. For extracellular recordings, 48-well MEAs were placed in a MAESTRO recording station and maintained at 37°C. Recordings were performed in DMEM / 10% heat-inactivated horse serum. The pH was maintained at 7.4 by a continuous flow of filtered, humidified air containing 10% CO2.

[0233] Each unit represents the activity arising from one neuron recorded with one electrode. Units were isolated at the beginning of the recording. For each unit, action potentials (i.e., spikes) were recorded as spike trains, a group of states called a "burst." Bursts were quantitatively described by direct spike train analysis using the programs Spike Wrangler and NPWaveX (both NeuroProof GmbH, Rostock, Germany). Bursts were defined by the onset and end of short spike events.

[0234] Multiparameter high-content analysis of network activity patterns extracted 204 activity-descriptive spike train parameters that allow for a precise description of activity changes in four categories: overall activity, burst structure, oscillatory behavior, and synchrony.

[0235] MPP for neuronal networks + The functional effects induced by the nanoparticles of the present invention and the preventive / remedial efficacy of the nanoparticles of the present invention were evaluated through the above parameters.

[0236] On day 21, values ​​related to spontaneous spontaneous activity were obtained from 60-second intervals obtained from a 30-minute time window 30 minutes after activity stabilization. Results (parameter values) were expressed as the mean ± SEM of independent networks. Each "nanoparticle" group, "control" group, and "MPP" group +For the "group," at least 19 active wells ("active" means wells with a sufficient number of electrodes to measure electrical activity) were included in the analysis. Absolute parameter distributions were tested for normality, and statistically significant differences between groups were assessed by one-way ANOVA.

[0237] To assess compound efficacy, we selected 204 parameters and expressed the multiparameter results in a single parameter called the "Effect Score." This is a linear combination of the selected features, and we divided the dataset into two groups: a "control" group with a mean value of "0" and an "MPP" group with a mean value of "1." + The calculation of the Z factor of the effect score was performed by selecting 20 features from the 204 parameters measured, and then dividing the result into a vector with the "control" group and the "MPP" group. + The z' factor was optimized to find the best discrimination between the 'groups' (Kuemmel A, et al. J Biomol Screen., 2010, 15(1), 95-101: Integration of multiple readouts into the z' factor for assay quality assessment).

[0238] The effect score analysis is shown in Figure 15.

[0239] The preventive / remedial efficacy of the nanoparticles of the present invention is shown in Table 7.

[0240] [Table 7]

[0241] As shown in FIG. 15 and Table 7, the MPP in the neuron network was improved by pre-processing the neuron network with the nanoparticles of the present invention. + Interestingly, gold nanoparticles with a population median maximum core size equal to 15 nm were found to induce MPPs in neuronal networks more effectively than gold nanoparticles with a population median maximum core size equal to 45 nm.+ Low efficiency in preventing / relieving induced functional effects.

[0242] These results suggest that the MPP in neuronal networks + The ability of both gold nanoparticles to prevent / relieve the induced functional effects is striking, with gold nanoparticles with a maximum median size of 45 nm being more efficient than gold nanoparticles with a maximum median size of 15 nm.

[0243] Example 11. Synthesis of nanoparticles prepared with conductive materials: poly(3,4-ethylenedioxythiophene) nanoparticles with negative surface charges (PEDOT nanoparticles) Poly(3,4-ethylenedioxythiophene) nanoparticle (PEDOT nanoparticle) dispersion in water (1.1 wt %) was obtained from Sigma (Sigma 675288) and used as received.

[0244] The zeta potential was determined by diluting the nanoparticle suspension in 1 mM NaCl solution at pH 7.3 (final PEDOT concentration: 1 g / L) and measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern). The zeta potential at pH 7.3 was found to be equal to -53 mV.

[0245] The median maximum dimension of the nanoparticles or nanoparticle aggregates in the population, as well as the size of the cores of nanoparticles or nanoparticle aggregates representing 30% to 70% of the population of nanoparticles and nanoparticle aggregates, were assessed using scanning electron microscopy (SEM) and were equal to 408 nm and 311 nm to 518 nm, respectively (56 nanoparticles were counted and their maximum dimension was measured).

[0246] Example 12. MPP using phenotypic MEA screening technology + Evaluation of the preventive / rescue efficacy of PEDOT nanoparticles from Example 11 in evoked neuronal networks The preventive / rescue efficacy of the nanoparticles of the present invention was evaluated by measuring the number of MPPs cultured on a 48-well MEA for 3 weeks. +The nanoparticles were tested in ventral midbrain / cortex co-cultures of treated mice. The preventive / rescue effects of the nanoparticles were assessed by measuring the extracellular electrical activity of neuronal co-cultures seeded on microelectrode array (MEA) chips.

[0247] In vitro induction of parkinsonian phenotype in mouse neurons using 1-methyl-4-phenylpyridinium iodide (MPP) + ) was carried out.

[0248] Materials and Methods Primary cell culture, treatment conditions Midbrain and frontal cortex tissues were collected from embryonic day 14.5 chr:NMRI mice (Charles River). Mice were sacrificed by cervical dislocation. Tissues were dissociated by enzymatic digestion (133.3 Kunitz units / ml DNase; 10 units / ml papain) and mechanical trituration, counted, and viability controlled. They were then seeded onto MEAs in 20 μl drops of DMEM containing laminin (10 μg / ml), 10% fetal bovine serum, and 10% horse serum. Cultures on MEAs were incubated at 37°C in a 10% CO atmosphere until ready for use. DMEM containing 10% horse serum was replenished twice weekly.

[0249] In the "nanoparticle" group, wells were treated with the nanoparticle suspension from Example 11 ([PEDOT] = 500 μM) on day 7, followed by 20 μM MPP + In the "control" group, water was added to the wells on the 7th day, followed by water on the 8th day. + In the " group, water was added to the wells on the 7th day, followed by 20 μM MPP on the 8th day. + Added.

[0250] MPP + (or water for the "control" group), 24 hours after addition, the medium was changed and MPP + After that, the medium was changed twice a week.

[0251] On day 21, 120 min of neuronal activity was recorded and 30 min of stable activity was analyzed.

[0252] Microelectrode Arrays and Neurochips 48-well microelectrode array neurochips were purchased from Axion Biosystems Inc. These chips contain 16 passive electrodes per well. The surfaces were coated with polyethyleneimine (PEI, 50% in borate buffer) for 1 hour, washed, and air-dried.

[0253] Multichannel recording and multiparameter data analysis Recordings were performed using a multichannel MAESTRO recording system from Axion Biosystems (USA). For extracellular recordings, 48-well MEAs were placed in a MAESTRO recording station and maintained at 37°C. Recordings were performed in DMEM / 10% heat-inactivated horse serum. The pH was maintained at 7.4 by a continuous flow of filtered, humidified air containing 10% CO2.

[0254] Each unit represents the activity arising from one neuron recorded with one electrode. Units were isolated at the beginning of the recording. For each unit, action potentials (i.e., spikes) were recorded as spike trains, a group of states called a "burst." Bursts were quantitatively described by direct spike train analysis using the programs Spike Wrangler and NPWaveX (both from NeuroProof GmbH, Rostock, Germany). Bursts were defined by the onset and end of short spike events.

[0255] Multiparameter high-content analysis of network activity patterns extracted 204 activity-descriptive spike train parameters that allow for a precise description of activity changes in four categories: overall activity, burst structure, oscillatory behavior, and synchrony.

[0256] MPP +The functional effects on neuronal networks induced by α-glucan and the preventive / remedial efficacy of the nanoparticles of the present invention were evaluated through the above parameters.

[0257] On day 21, values ​​related to spontaneous spontaneous activity were obtained from 60-second intervals obtained from a 30-minute time range after 30-90 minutes of activity stabilization. Results (parameter values) were expressed as the mean ± SEM of independent networks. For the "nanoparticle" group, at least 5 active wells were required; for the "control" group, at least 20 active wells were required; for the "MPP" group, at least 10 active wells were required. + For the "group," at least 20 active wells ("active" means wells with a sufficient number of electrodes to measure electrical activity) were included in the analysis. Absolute parameter distributions were tested for normality, and statistically significant differences between groups were assessed by one-way ANOVA.

[0258] To assess compound efficacy, the multiparameter result of the selection of 204 parameters was expressed in a single parameter called the "Effect Score." This is a linear combination of the selected features, dividing the dataset into a "Control" group with a mean value of "0" and a "MPP" group with a mean value of "1." + The Z-factor of the effect score is calculated through feature selection of 20 out of 204 parameters, and the vector is converted into a vector with a "control" group and a "MPP" group. + The z' factor was optimized to find the best discrimination between the 'groups' (Kuemmel A, et al. J Biomol Screen., 2010, 15(1), 95-101: Integration of multiple readouts into the z' factor for assay quality assessment).

[0259] The effect score analysis is shown in Figure 17.

[0260] The preventive / remedial efficacy of the nanoparticles of the present invention is shown in Table 8.

[0261] [Table 8]

[0262] As shown in FIG. 17 and Table 8, pretreatment of the neuronal network with the PEDOT nanoparticles of the present invention significantly improved the MPP in the neuronal network. + It is shown that the induced functional effect is prevented / relieved.

[0263] These results suggest that MPP in neuronal networks + This highlights the ability of the nanoparticles described herein to prevent / relieve induced functional effects.

[0264] Example 13. Synthesis of nanoparticles prepared by insulating materials with low dielectric constants below 100: Synthesis of hafnium oxide nanoparticles coated with a biocompatible coating having a negative surface charge Hafnium oxide (HfO2) nanoparticles were synthesized by precipitating hafnium chloride (HfCl4) with tetramethylammonium hydroxide (TMAOH) at basic pH. The resulting suspension was transferred to an autoclave and heated at temperatures above 110 °C. After cooling, the suspension was washed with deionized water and acidified.

[0265] Surface functionalization was performed using sodium hexametaphosphate. A sufficient amount of sodium hexametaphosphate was added to the nanoparticle suspension to reach at least half a monolayer coverage on the surface (2.5 molecules / nm 2 The nanoparticle suspension was stirred overnight and subsequently the pH was adjusted to 7.

Claims

1. 1. A nanoparticle or nanoparticle aggregate for use in the prevention or treatment of a neurological disorder or at least one symptom thereof in a subject, without exposing the nanoparticle or nanoparticle aggregate to an electric field or any other external activation source, comprising: The material of the nanoparticles or nanoparticle aggregates is a conductive material, a semiconductive material, or a material having a dielectric constant ε of 200 or more. ｉｊｋ and an insulating material having a dielectric constant ε of 100 or less. ｉｊｋ and the insulating material is selected from the group consisting of where: i) the maximum median size for a population of cores of said nanoparticles or nanoparticle aggregates is such that the material is a conducting material, a semiconducting material or a material with a dielectric constant ε of 200 or greater; ｉｊｋ at least 30 nm when the insulating material has ii) nanoparticles or nanoparticle aggregates, the core of which is coated with a biocompatible coating that provides a neutral or negative surface charge when measured in an aqueous solution having an electrolyte concentration of 0.001-0.2 M, a nanoparticle or nanoparticle aggregate material concentration of 0.01-10 g / L, and a pH of 6-8.

2. 2. The nanoparticles or nanoparticle aggregates for use according to claim 1, wherein the material of the nanoparticles or nanoparticle aggregates is a conductive material selected from metals having a standard reduction potential E° greater than 0.2 and organic materials having consecutive sp2 hybridized carbon centres in their structure.

3. 3. The nanoparticles or nanoparticle aggregates for use according to claim 2, wherein the material of the nanoparticles or nanoparticle aggregates is selected from metal nanoparticles whose metallic element is Ir, Pd, Pt, Au or a mixture thereof, and organic nanoparticles made of polyaniline, polypyrrole, polyacetylene, polythiophene, polycarbazole and / or propylene.

4. 2. The nanoparticle or nanoparticle aggregate for use according to claim 1, wherein the material of the nanoparticle or nanoparticle aggregate is a semiconductor material having a band gap Eg below 3.0 eV.

5. 5. The nanoparticles or nanoparticle aggregates for use according to claim 4, wherein the material of the nanoparticles or nanoparticle aggregates consists of elements from group IVA of the Mendeleev periodic table, or a mixed composition of elements from groups III and V of the Mendeleev periodic table, or a mixed composition of elements from groups II and VI of the Mendeleev periodic table.

6. 6. The nanoparticles or nanoparticle aggregates for use according to claim 5, wherein the material of the nanoparticles or nanoparticle aggregates consists of an element from group IVA of the Mendeleev periodic table and is doped with a charge carrier selected from Al, B, Ga, In and P.

7. The material is an insulating material having a band gap Eg of 3.0 eV or more, and a relative dielectric constant ε ｉｊｋ However, 20℃ to 30℃ and 10 ２ 2. The nanoparticles or nanoparticle aggregates for use according to claim 1, measured between 100 Hz and infrared frequencies.

8. The material is an insulating material having a band gap Eg of 3.0 eV or more, and a relative dielectric constant ε ｉｊｋ is 200 or more, and the material of the nanoparticles or nanoparticle aggregates is BaTiO ３ , KTaNbO ３ , KTaO ３ , SrTiO ３ and BaSrTiO ３ 8. The nanoparticles or nanoparticle aggregates for use according to claim 7, wherein the dielectric material is a mixed metal oxide selected from:

9. The material is an insulating material having a band gap Eg of 3.0 eV or more, and a relative dielectric constant ε ｉｊｋ is 100 or less, and the material of the nanoparticles or nanoparticle aggregates is ReO ２ , ZrO ２ and HfO ２ 8. The nanoparticles or nanoparticle aggregates for use according to claim 7, which are metal oxides selected from:

10. 10. The nanoparticles for use according to any one of claims 1 to 9, wherein the neurological disease is selected from Parkinson's disease, Alzheimer's disease, epilepsy, obsessive-compulsive disorder, autism spectrum disorder, depressive disorder, dystonia, Tourette's syndrome, schizophrenia, stroke, aphasia, dementia, tinnitus, Huntington's disease, essential tremor, bipolar disorder, anxiety disorder, addiction disorder, consciousness vegetative state, and at least one symptom thereof.

11. A composition comprising the nanoparticles and / or nanoparticle aggregates according to any one of claims 1 to 9 and a pharmaceutically acceptable support, The composition is for use in preventing or treating a neurological disorder or at least one symptom thereof in a subject without exposing the nanoparticles and / or nanoparticle aggregates to an electric field or any other external activation source.

12. The composition for use according to claim 11, wherein the composition comprises at least two different nanoparticles and / or nanoparticle aggregates according to any one of claims 1 to 9.

13. A kit comprising at least two different nanoparticles and / or nanoparticle aggregates according to any one of claims 1 to 9.

14. 14. The kit of claim 13, for use in preventing or treating a neurological disorder or at least one symptom thereof in a subject without exposing the nanoparticles and / or nanoparticle aggregates to an electric field or any other external activation source.

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