Nanoparticles for use in treating nerve disorders

Abstract

We provide medical services, particularly treatment for neurological disorders. [Solution] 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 when exposed to an electric field, wherein the material of the nanoparticles or nanoparticle aggregates is a conductive material, a semiconductor material, and has a dielectric constant ε of 200 or more. ijk An insulating material having a dielectric constant ε of 100 or less ijk The present invention provides nanoparticles or nanoparticle aggregates selected from insulating materials having [a certain property]. Furthermore, the present invention provides compositions and kits comprising such nanoparticles and / or nanoparticle aggregates, as well as uses thereof.

Description

[Technical Field]

[0001] The present invention relates to the medical field, particularly 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 when the nanoparticles or nanoparticle aggregates are exposed to an electric field / electrical stimulation, wherein the material of the nanoparticles or nanoparticle aggregates is a conductive material, a semiconductor material, and has a dielectric constant ε of 200 or more. ijk An insulating material having a dielectric constant ε of 100 or less ijk The present invention relates to nanoparticles or nanoparticle aggregates selected from insulating materials having [a certain property]. Furthermore, the present invention relates to compositions and kits comprising such nanoparticles and / or nanoparticle aggregates, and to the use thereof. [Background technology]

[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 motor disorder caused by the death of dopamine neurons in the substantia nigra located in the midbrain. Stroke corresponds to the disruption of blood supply to the brain. Without oxygen, neurons in the affected area die, and the parts of the body controlled by these cells become unable to function. Huntington's disease is a genetic disorder. Epilepsy is a disorder caused by the abnormal excitation of large 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 disorder are multifactorial, including genetic and environmental factors.

[0003] Neurological disorders can be classified according to the main symptoms affecting the patient. Three main types of symptoms are observed: motor disorders, psychiatric (mood / social) disorders, and cognitive disorders, as will be further described below in this specification.

[0004] Movement disorders include tremors, reduced motor function, such as bradykinesia or dyskinesia, muscle torsion, rigidity, postural instability, and foot dystonia. Diseases presenting with movement disorders typically include Parkinson's disease, dystonia, epilepsy, Huntington's disease, and Tourette syndrome.

[0005] Mental disorders constitute various diseases presenting with symptoms of mood / social disorders. A non-exhaustive list includes autism spectrum disorder, schizophrenia spectrum disorder, bipolar disorder, depressive disorder, anxiety disorder, obsessive-compulsive disorder, substance-related and / or addictive disorders (defined from the Diagnostic and Statistical Manual of Mental Disorders, 2013, 5th edition, the American Psychiatric Association). Among patients with movement disorders, such as Parkinson's disease and dystonia, mental disorders may develop in the later stages of the disease.

[0006] Cognitive impairment exists in many, if not all, mental disorders (e.g., schizophrenia, bipolar disorder). Only disorders with a central cognitive feature are included in the cognitive impairment category. Cognitive impairment affects a patient's daily life and makes it difficult to achieve simple tasks. Dementia is a typical cognitive impairment and is a general term for a severe decline in mental ability that interferes with daily life. Alzheimer's disease is a specific type of dementia with a neurodegenerative aspect.

[0007] Neurological disorders are treated, where possible, with drugs that play a role in regulating neurotransmitter levels in the brain and controlling their interactions with specific neurotransmitter receptors. The main neurotransmitters involved are glutamates, gamma-aminobutyric acid (GABA), dopamine, and acetylcholine. Glutamates and GABA neurotransmitters are of particular interest because they play major roles in increasing (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 (Holmes et al., Mental Retardation and Developmental Disabilities, 1995, 1, 208-219: Role of glutamate and GABA in the pathophysiology of epilepsy) neuronal excitability, respectively. Dopamine is involved in several brain functions: motor control via the basal ganglia (inappropriate levels of dopamine in the basal ganglia can lead to uncontrolled movement), pleasure-seeking behavior (impairment can lead to dysfunctional addiction), and cognition (impairment of dopamine in the frontal lobe can lead to a decline in neurocognitive function) (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] A common drug for reducing the motor symptoms of Parkinson's disease is levodopa, which is converted into dopamine in the brain and thus helps to balance the dopamine deficiency. Levodopa is related to carbidopa, which helps to avoid the conversion of levodopa into dopamine throughout the body. One problem with levodopa treatment is the "on-off" phenomenon, which results in stages of immobility and incapacity related to depression that alternate with a jubilant thaw (Lees et al., J Neurology Neurosurgery Psychiatry, Special Supplement, 1989, 29-37: The on-off phenomenon). The unresponsiveness of patients with late-stage Parkinson's disease to this treatment is a problem (Fabbri et al., Parkinsonism and related disorders, 2016: Do patients with late-stage Parkinson’s disease still respond to levodopa?). Other common drugs for treating the "positive" symptoms in schizophrenia, symptoms of neuropsychiatric disorders such as delusions and hallucinations, are antipsychotics. <> <>

[0009] <> However, the therapeutic treatment of drug-induced neurological symptoms is non-specific and can induce severe adverse events. In addition, treatment resistance to the drugs used can appear. <>

[0010] ​​As our understanding of neuroscience advances, the brain can be thought of as an electrical network that encodes and transmits information through its "wires," the neurons. The connectivity between neurons is both simple and complex. It is simple because the influx / outflow of ions within neurons generates action potentials (or "spikes" of electrical activity). It is complex because the brain network consists of hundreds of billions of neurons that 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 (structure) that connect individual neurons and the processes (function) that transmit information. Both aspects influence the overall performance of the nervous system. Neural interactions are induced by oscillations in brain electrical activity patterns, which are typically measurable by electroencephalography (EEG). Different frequency bands: delta, theta, alpha, beta, and gamma oscillations are observed (Ward et al., Trends in Cognitive Sciences, 2003, 7(12), 553-559: Synchronous neural oscillations and cognitive processes). Structurally, the most prominent neuroanatomical feature of the brain is the rich connectivity between neurons, which reflects the importance of neurotransmission. It is thought that the synchronization of oscillations between one brain region and another ("synchronization") leads to the final level of information coding [first level (neurons): action potentials; second level (neuronal networks): neuronal oscillations], resulting in spatiotemporal coordination (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 a delicately balanced pattern of synchronization and desynchronization in space and time is 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 (synchronization that is too high and / or too long (i.e., also called hypersynchronization) or too low (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, the 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. Certain neurological disorders affect deep brain regions where the penetration depth of the electric field is weak; therefore, surgical implantation of electrodes in the brain to continuously deliver electrical stimulation is performed, constituting the "deep brain stimulation" (DBS) technique. Its effectiveness depends on the parameters used for stimulation, particularly the frequency. In 1987, it was found that high-frequency stimulation (≧100Hz) of the ventralis intermedius (VIM) with implanted electrodes reduced 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 disease). Furthermore, in monkeys, high-frequency stimulation (>100Hz) compared to low-frequency stimulation (<50Hz) enables changes in the temporal firing patterns of neurons in the external segment (GPe) and internal segment (GPi) of the globus pallidus (GPi), resulting in a regular firing pattern synchronized with the stimulus. This change blocks the transmission of altered neural activity patterns in the basal ganglia to their target structures in the thalamus and brainstem, thus reducing bradykinesia and rigidity (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 to treat several motor 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 procedure and the risk of various complications, such as bleeding, epileptic 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 focalness (i.e., spatial resolution) of the electric field generated within the target is another concern. The spread of electrical stimulation is also associated with side effects such as depression. Much research has focused on designing novel types of electrodes that can shift and confine stimulation within specific regions (Luan et al., Frontiers in Neuroengineering, 2014, 7(27), 1-9: Neuromodulation: present and emerging methods). Other technical aspects—electrodes (or leads), their size, the invasiveness of DBS devices, the materials constituting the leads, compatibility with (magnetic resonance) imaging techniques, and the battery life of internal pulse generators (IPGs) related to the need for continuous stimulation—are under evaluation.

[0015] Other major existing types of electrical stimulation, namely 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 applications are limited to stimulating the cerebral cortex (they cannot reach deep brain tissue). Furthermore, their spatial resolution remains low.

[0016] Electrical stimulation of the brain remains a relevant method for treating neurological disorders. However, to avoid side effects, such as psychiatric side effects, and ultimately to improve the benefit / risk ratio of the treatment, it is necessary to increase the localized delivery of electrical stimulation and the penetration depth without affecting surrounding brain regions.

[0017] In recent years, non-invasive neurostimulation techniques, such as the use of light or ultrasound to directly stimulate neurons, have been considered. However, these techniques have the drawback of low spatial resolution.

[0018] Interestingly, nanomaterials with unique properties have been explored as mediators for converting wirelessly transmitted primary stimuli into localized secondary stimuli, primarily electric fields or heat, at nanomaterial-neuron interfaces (Wang Y. & Guo L. Frontiers in Neuroscience. 2016; vol. 10, Article 69, Nanomaterial-enabled neural stimulation). For this reason, photoelectric conversion using quantum dots, photothermal conversion using gold nanomaterials, magnetoelectric conversion using magnetoelectric nanoparticles, magnetothermal conversion using superparamagnetic nanoparticles, and acoustic-electroacoustic conversion using piezoelectric nanomaterials have been demonstrated.

[0019] Most of these emerging technologies using nanomaterials require the simultaneous development of an energy source to deliver nerve stimulation. Furthermore, the incoming energy needs to be converted into efficient secondary stimulation that requires well-defined nanoparticle structure and composition, as well as the persistence of the nanoparticle structure and composition over time.

[0020] For example, magnetoelectric (ME) nanoparticles are composite nanoparticles that exhibit piezoelectric and magnetostrictive properties. Specifically, the ME effect enabled by, for example, CoFe2O4-BaTiO3 nanoparticles arises from the combined action of two distinct materials, namely, a magnetostrictive (CoFe2O4) material and a piezoelectric (BaTiO3) material. More precisely, when CoFe2O4-BaTiO3 nanoparticles are exposed to a magnetic field, firstly, the magnetostrictive material generates local stress due to a change in its length (volume), and secondly, the piezoelectric material generates polarization (charge) in response to this local stress. Neither the magnetostrictive material nor the piezoelectric material can generate either the ME effect or polarization on their own when exposed to a magnetic field, as explained by Grossinger R. et al. (Grossinger R. et al., Journal of Magnetism and Magnetic Materials, 2008, 320, 1972-1977: The physics of magnetoelectric composites). [Overview of the project] [Means for solving the problem]

[0021] The present invention deals with nanoparticles and / or nanoparticle aggregates (aggregates of nanoparticles) for use in the prevention or treatment of neurological disorders (typically disorders of neuronal networks) or at least one symptom thereof when nanoparticles or nanoparticle aggregates are exposed to an electric field. The electric field is typically applied by deep brain stimulation (DBS), transcranial electrical stimulation (TES), or transcranial magnetic stimulation (TMS).

[0022] Nanoparticles or nanoparticle aggregates normalize (improve synchronization) the synchronization of neuronal oscillations within and / or between neuronal networks, and within and / or between separate regions of the brain, thereby increasing the spatial resolution (focus) of electrical stimulation while using standard electrical stimulation techniques. For this reason, the nanoparticles or nanoparticle aggregates described herein by the inventors help the subject / patient return to a healthy state.

[0023] Furthermore, since the nanoparticles or nanoparticle aggregates of the present invention enable a reduction in the applied current, voltage, pulse width, and / or frequency, known potential toxicity associated with applied / induced current is reduced.

[0024] Simple explanation In this specification, the use of nanoparticles or nanoparticle aggregates for the prevention or treatment of neurological disorders or at least one symptom thereof in subjects requiring prevention or treatment, when the nanoparticles or nanoparticle aggregates are exposed to an electric field / electrical stimulation, excited or activated by an electric field / electrical stimulation, is described for the first time as advantageous. The materials of the nanoparticles or nanoparticle aggregates are typically conductive materials, semiconductor materials, and materials with a dielectric constant ε of 200 or more. ijk An insulating material having a dielectric constant ε of 100 or less ijk An insulating material having the following properties is selected.

[0025] Furthermore, this specification also describes the use of nanoparticles or nanoparticle aggregates for preparing compositions for preventing or treating neurological diseases or at least one symptom thereof described herein in subjects requiring prevention or treatment.

[0026] Furthermore, in this specification, a composition for use in preventing or treating neurological disorders or at least one symptom thereof in an object exposed to an electric field, wherein the composition comprises or consists of nanoparticles and / or nanoparticle aggregates and a pharmaceutically acceptable support, wherein the material of the nanoparticles or nanoparticle aggregates is typically a conductive material, a semiconductor material, or a dielectric constant ε of 200 or more. ijk An insulating material having a dielectric constant ε of 100 or less ijk Compositions selected from insulating materials having the property are also described.

[0027] Furthermore, this specification includes at least two distinct nanoparticles and / or nanoparticle aggregates, each nanoparticle or nanoparticle aggregate being a conductive material, a semiconductor material, and having a dielectric constant ε of 200 or more. ijk An insulating material having a dielectric constant ε of 100 or less ijk A kit comprising separate materials typically selected from insulating materials having a property, and its use typically in the prevention or treatment of neurological disorders or at least one symptom thereof in a subject / in a method for preventing or treating them. [Modes for carrying out the invention]

[0028] Detailed explanation 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 characteristic of a neuron (or nerve cell) is its ability to transmit electrical signals in the form of action potentials.

[0029] Neurons / nerve cells constitute the basic nodes of the brain. Nerve cells can communicate with each other in a highly structured manner to form neural networks. Neurons communicate via synaptic connections. Within neurons, nanocircuits constitute the underlying biochemical mechanisms for mediating important neuronal properties, such as learning and memory, and the generation of neuronal rhythms.

[0030] Microcircuits can be formed by just a few interconnected neurons and can mediate complex tasks such as mediating reflexes, processing sensory information, initiating movement, and learning and memory. Macrocircuits are more complex networks consisting of multiple embedded microcircuits. Macrocircuits mediate higher brain functions, such as object recognition and perception. Thus, multiple levels of networks occupy the nervous system.

[0031] Excitability of neural networks Neurons transmit messages electrochemically (i.e., chemicals / ions produce electrical signals). Important ions in the nervous system are sodium and potassium, calcium, and chloride. When a neuron is not transmitting signals, it is in a "resting state." In the resting state, the inside of the neuron is negative relative to the outside. Attempts are made to balance the concentrations of various ions on both sides of the membrane, but balance is not achieved. This is 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 called the "resting potential") of the neuron is approximately -70mV. This means that the inside of the neuron is 70mV lower than the outside. In the resting state, there are relatively more sodium ions outside the neuron and more potassium ions inside that neuron. An action potential (also identified as a "spike" or "impulse") is generated when a neuron detaches from its cell body and travels down its axon, transmitting information. This means that some event (stimulus) causes the resting potential to move towards 0 mV. When depolarization reaches approximately -55 mV, the neuron fires an action potential. If depolarization does not reach this critical threshold level, the action potential does not fire (on / off mechanism). Also, once the threshold level is reached, a fixed-amplitude action potential always fires. Therefore, either depolarization does not reach the threshold or a complete action potential is generated.

[0032] The propagation speed of action potentials exhibits considerable variability. In fact, the propagation speed of action potentials in nerves can vary from 100 meters per second to less than one-tenth of a meter per second. The time constant is an indicator of how quickly the membrane responds to a stimulus in time, while the spatial constant (also called the length constant) is an indicator of how well the potential spreads along the axon as a function of distance.

[0033] Connectivity within and between neuronal networks Three types of connectivity networks exist that are used to investigate transmission within and across the brain. Structural connectivity is based on the detection of fiber tracks that physically connect regions of the brain. These are anatomical network maps that show the possible pathways through which signals can travel within the brain. Functional connectivity identifies activity in brain regions that have similar frequencies, phases, and / or amplitudes of correlated activity. Effective connectivity uses functional connectivity information and goes a step further to determine the direct or indirect influence that one nervous system may 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).

[0034] Synchronized activity within neuronal networks can be detected by imaging techniques such as magnetoencephalography (MEG), electroencephalography (EEG), functional magnetic resonance imaging (FMRI), or positron emission tomography (PET), followed by network connectivity analysis. 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. Certain regions within the brain are specialized for processing specific types of information. Imaging techniques have revealed that these regions are connected and communicate across networks within the brain to other specialized regions. 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 synchronicity (synchronized or synchronized state) of neuronal patterns in oscillating brain activity. Detecting neuronal synchronous activity can be used to determine the health or completeness 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 can help evaluate treatment based on pre- and post-treatment brain connectivity imaging.

[0035] An intact brain exhibits a complex pattern of synchronous activity related to different "states" of the organism, from slow delta rhythms (0.5 - 4 Hz) to theta (4 - 8 Hz), alpha (8 - 12 Hz), beta (15 - 30 Hz), and gamma (30 - 70 Hz) oscillations. Interestingly, dissociated cultures of cortical structures provide a convenient system for testing the rules that govern the emergence, generation, and spread of network firing (spikes) and bursts (clusters of spikes) in populations of densely interconnected neurons. Network activity can be recorded over long periods with finite temporal resolution using multi - electrode arrays in a non - invasive manner. Two - dimensionally separated cultures can be used as a feasible test system for studying the rules that control the formation and maintenance of network activity in the brain, enabling the testing of hypotheses that cannot be addressed in an intact brain (Cohen E. et al., Brain Research, 2008, 1235, 21 - 30: Determinants of spontaneous activity in networks of cultured hippocampus).

[0036] As used herein, the use of nanoparticles or nanoparticle aggregates for preventing or treating a neurological disorder or at least one symptom thereof in a subject in need of prevention or treatment when the nanoparticles or nanoparticle aggregates are exposed to an electric field / for use in such prevention or treatment is advantageously described for the first time. The material of the nanoparticles or nanoparticle aggregates is typically selected from conductor materials, semiconductor materials, insulator materials having a dielectric constant ε of 200 or more and insulator materials having a dielectric constant ε of 100 or less. ijk and having a dielectric constant ε of 100 or less. ijk is selected from insulator materials having a dielectric constant ε of 100 or less.

[0037] The term “treatment” refers to any therapeutic treatment or means that can prevent, alleviate, or cure any of the diseases, disorders, or dysfunctions described herein. Such treatments are intended for mammalian subjects, preferably human subjects, who require such treatment. Thus, there may be already identified (diagnosed) subjects suffering from any of the diseases, disorders, or dysfunctions described herein, or subjects who are considered to be at risk of developing such diseases, disorders, or dysfunctions, for which the treatment is preventive or prophylactic.

[0038] Abnormal modulation of interneuronal oscillatory is actually present in different types of neurological disorders or conditions (also referred to herein as “neural disorders or conditions”) (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).

[0039] The human nervous system is divided into the central nervous system (CNS) and the peripheral nervous system (PNS). The CNS is further divided into the brain and spinal cord, located in the cranial cavity and spinal canal of the skull, respectively. The CNS and PNS work together to integrate sensory information and control motor and cognitive functions. Figure 1 shows a simplified diagram of the brain structure.

[0040] Synchronization (or synchronization) within and between neuronal networks within and / or between distinct brain regions occurs temporally through the coordination of neuronal oscillations (Buzsaki et al., Science, 2004, 304, 1926-1929: Neuronal oscillations in cortical networks). Motor disorders are typically due to hypersynchronization, which means that the synchronization of oscillations within and / or between neuronal networks within and / or between distinct brain regions is too high and / or too long. Mental and cognitive disorders are typically due to dyssynchrony, which means that the synchronization of oscillations within and / or between neural networks in separate brain regions and / or between separate brain regions is reduced (typically showing decreased activity) or even absent (see Table 1: Abnormal Neural Synchrony in Neurological Disorders (adopted from Uhlhaas et al., Neuron, 2006, 52, 155-168: Neural synchrony in brain disorders: relevance for cognitive dysfunctions and pathophysiology)).

[0041] [Table 1]

[0042] Since "coherence" is a mathematical technique that quantifies the frequency and amplitude of the synchronous nature (synchronized or synchronized state) of neuronal patterns in oscillating brain activity, excessively high and excessively low coherence can be thought to be associated with 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).

[0043] In certain embodiments, the neurological disorders or conditions targeted in the context of the present invention are selected from Parkinson's disease, Alzheimer's disease, epilepsy, obsessive-compulsive disorder, autism spectrum disorder, depressive disorder, dystonia, Tourette syndrome, schizophrenia, stroke, aphasia, dementia, tinnitus, Huntington's disease, essential tremor, bipolar disorder, anxiety disorder, addiction disorder, and consciousness vegetative state, for example, selected from Parkinson's disease, Alzheimer's disease, epilepsy, obsessive-compulsive disorder, autism spectrum disorder, depressive disorder, dystonia, Tourette syndrome, schizophrenia, stroke, aphasia, dementia, tinnitus, Huntington's disease, essential tremor, bipolar disorder, addiction disorder, consciousness vegetative state, and at least one of these conditions.

[0044] As already described above in this specification, neurological disorders or disorders can be classified according to the main symptoms affecting the patient, which are motor disorders, mental (mood / social) disorders, and cognitive disorders, as will be further detailed below in this specification.

[0045] Examples of motor impairments Parkinson's disease Parkinson's disease (PD) affects approximately 7 to 10 million people worldwide and is characterized by tremors, dyskinesia, bradykinesia, and freezing of gait. PD is a slowly progressing degenerative disease of the brain. PD affects nerve cells in brain regions called the basal ganglia and the substantia nigra. Nerves in the substantia nigra produce dopamine, a neurotransmitter that acts as a chemical messenger in brain circuits crucial for planning and controlling bodily movements. In PD, dopamine-producing nerve cells 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). If dopamine receptors in the striatum are not sufficiently stimulated, some parts of the basal ganglia become either understimulated or overstimulated. In particular, the subthalamic nucleus (STN) becomes hyperactive and acts as a facilitator on the globus pallidus internal segment (GPi). Excessive stimulation of the GPi has an excessive inhibitory effect on the thalamus, which in turn reduces its output, leading to deceleration and rigidity of movement (Guo et al., Frontiers in Computational Neuroscience, 2013, 7, 124, 1-11: Basal ganglia modulation of thalamocortical relay in Parkinson's disease and dystonia).

[0046] Dopamine deficiency in Parkinson's disease (PD) is associated with excessive oscillation synchronization at beta frequencies throughout the corticobasal ganglia motor network. Indeed, dopamine levels in the basal ganglia are predicted to suppress beta synchronization, which in turn mediates the dopaminergic involvement necessary for motor prediction (Jenkinson et al., Trends in Neuroscience, 2011, 34(12), 611-618: New insights into the relationship between dopamine, beta oscillations and motor function). If dopamine levels in the basal ganglia are not sufficiently high, beta oscillation synchronization may no longer be controlled, potentially resulting in slower movement. Another observation in Parkinson's disease patients leads to the conclusion that cortical oscillations in the beta band induce and drive cortical 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).

[0047] 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). Treatment of PD symptoms with DBS has been approved by the FDA since 2002 (and since 1997 for essential tremor). In combination with the nanoparticles described herein, the most commonly used stimulation parameters available in the context of this invention are a frequency of 130–185 Hz, a pulse width of 60–210 μs, and a voltage amplitude of 1–3.5 V (Kuncel et al., Clinical Neurophysiology, 2004, 115, 2431-2441: Selection of stimulus parameters for DBS). 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 transcranial stimulation of the cortex (e.g., transcranial magnetic stimulation - TMS) can also be used to treat the symptoms of Parkinson's disease (Cantello et al., Brain Research Reviews, 2002, 38, 309-327: Transcranial magnetic stimulation and Parkinson's disease).

[0048] Dystonia Dystonia is a neurological disorder characterized by abnormal, involuntary twisting and rotational movements that reflect impaired motor function. Several forms of dystonia exist, depending on the affected body part, their genetic origin, and the types of neurotransmitters involved. The central nervous system (CNS) in dystonia exhibits incomplete inhibition, leading to a loss of reciprocal vertebral inhibition between opposing muscles. For example, in upper limb dystonia, abnormal synchronization of neurons / nerves that provide input signals to antagonistic muscles in the forearm results in co-contraction of these antagonistic muscles (the symptoms 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).

[0049] An interesting DBS target point showing anti-dystonia effects is the globus pallidus internal segment (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). The effect of stimulating the ventral intermediate nucleus (VIM-DBS) of the thalamus is much weaker. Stimulation using the subthalamic nucleus (STN-DBS) has been experimental. GPi-DBS provides relief from the main symptoms of dystonia, but it may take several weeks to several months for the therapeutic effect to fully manifest (Dressler et al., J Neural Transm, 2015, DOI 10.1007 / s00702-015-1453-x: Strategies for treatment of dystonia). In combination with the nanoparticles described herein, the most commonly used stimulation parameters available in the context of the present invention are a frequency of 130–180 Hz, a pulse width of 60–210 μs, and an amplitude of 2–5 volts.

[0050] epilepsy Epilepsy is a brain disorder affecting approximately 50 million people worldwide, primarily characterized by recurrent and unpredictable interruptions of normal brain function called epileptic seizures. Epilepsy is not a single disease entity, but rather a range of disorders that reflect underlying brain dysfunction that can result from many different causes (genetic mutations, brain tumors, head trauma, stroke, alcoholism, brain inflammation; meningitis, infections such as HIV or viral encephalitis) (Fisher et al., Neurology, 2015, 28(2), 130-135: Redefining epilepsy). Epileptic seizures are defined as the transient occurrence of signs and / or symptoms resulting from 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 medial temporal lobe seizures (National Institute of Neurological Disorders and Stroke: http: / / www.ninds.nih.gov / disorders / epilepsy / detail_epilepsy.htm#3109_7). Identifying elevated local EEG synchrony regions or "hypersynchrony" regions in the cortex suggests that local hypersynchrony may be a marker for seizure-prone areas (Schevon et al., Neuroimage, 2007, 35(1), 140-148: Cortical abnormalities in epilepsy revealed by local EEG synchrony).

[0051] Neurostimulation for the treatment of epilepsy can take the form of peripheral nerve stimulation, such as vagus nerve stimulation (VNS), spinal cord stimulation, transcranial-to-brain stimulation (TES or TMS), or deep brain stimulation (DBS). Responsive neurostimulation is another strategy. In this case, stimulation is delivered only when seizure onset is detected. In 2004, a proof-of-principle study of responsive neurostimulation was published in three epilepsy patients, two of whom were treated with cortical grid or strip electrodes, and one with deep hippocampal electrodes. Individual seizures could be terminated at the onset of stimulation, and the overall seizure frequency was reduced by 50–75% (Kossoff et al., Epilepsia, 2004, 45, 1560-1567: Effect of an external responsive neurostimulator on seizures and electromagnetic discharges during subdural electrode monitoring). 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). A multicenter, randomized, controlled trial of bilateral stimulation of the anterior thalamic nucleus (SANTE) for epilepsy was conducted in 110 adult patients with partial seizures with or without secondary generalization occurring at least 6 times per month, but ≤10 times per day. Baseline seizure frequency was recorded for 3 months, followed by DBS lead implantation, 1 month of recovery, and then a 3-month blinded period of either active stimulation or no stimulation (placebo). The on-stimulation parameter was a 90 μs pulse at 145 Hz with 5 V for 1 minute, followed by 5 minutes without stimulation. Seizure frequency decreased by a median of 20% from baseline during the 1-month recovery period. Subsequently, the seizure frequency clearly differed between the two treatment groups, with a median improvement of 40.4% in the active group and 14.5% in the placebo group.In the active group, complex partial seizures were significantly less frequent, as were seizures of the type pre-designated as "most severe" by the patients (Fisher et al., Epilepsia, 2010, 51, 899-908: Electrical stimulation of the anterior nucleus of thalamus for treatment of refractory epilepsy). The electrical stimulation / treatment conditions described herein can be used in combination with the nanoparticles described herein in the context of the present invention.

[0052] Examples of mental disorders (mood and social disorders) Obsessive-compulsive disorder (OCD) Obsessive-compulsive disorder (OCD) is a common mental disorder that is often chronic, severe, and extremely debilitating. Furthermore, OCD is usually refractory to treatment, with a significant percentage of patients either not responding or experiencing only partial relief.

[0053] Functional neuroimaging studies have demonstrated dysfunction in the orbitofrontal cortex, basal ganglia, and striatum.

[0054] One study showed that acute OCD symptoms may be associated with abnormally high-oscillating 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 firing rate when suspicion arose during checking (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).

[0055] Deep brain stimulation (DBS) of the ventral forelimb (VC) and adjacent ventral striatum (VS) of the internal capsule has been approved in the EU for the treatment of severe and highly treatment-resistant OCD (VC / VS-DBS). To demonstrate the therapeutic prospects of this technique, four clinical centers collaborated most closely over eight years in a small study, and their data were analyzed (Greenberg et al., Molecular Psychiatry, 2010, 15, 64-79: Deep brain stimulation of the ventral internal capsule / ventral striatum for obsessive-compulsive disorder: worldwide experience). DBS leads were implanted bilaterally to stimulate the dorsal-ventral range of the anterior capsule. The electrical stimulation frequency was 100–130 Hz, and the pulse width ranged from 90 to a maximum of 450 μs. The DBS intensity was generally in the range of 2–8 V, resulting in a current ranging from approximately 2–15 mA, depending on the electrode impedance (generally 500–1000 Ω). The conclusion from these combined data is that clinically significant symptom reduction and functional improvement were observed in approximately two-thirds of highly treatment-resistant patients (26 patients in total), which gives hope for the therapeutic effects after VC / VS-DBS. The electrical stimulation / treatment conditions described herein can be used in combination with the nanoparticles described herein in the context of the present invention.

[0056] Autism Spectrum Disorder Autism is a neurodevelopmental disorder characterized by deficits in social interaction and communication, and abnormally restricted, repetitive behaviors. Autism typically begins in infancy, or at the latest within the first three years of life. Autism is a heterogeneous condition (two children or adults with autism will not have the same profile), and this condition gives rise to the concept of "autism spectrum disorder," which classifies the disorder into several levels according to the degree of language deficit or overall 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 this spectrum, individuals with autism are highly functioning and capable of living independently and maintaining employment. Individuals characterized as low-functioning exhibit more severe symptoms: difficulties with language (or even non-verbal communication), poor social communication, self-injurious behavior (SIB), tantrums, and potentially life-threatening aggression. A key trend in structural and functional brain studies of autism is the involvement of networks for socio-emotional processing: the limbic system, facial processing system, and mirror neuron networks. Deficiencies in gamma-band oscillation synchronization have been shown to be involved in the manifestation of symptoms (Sinha et al., Neurosurgery Focus, 2015, 38(6), E3: Deep brain stimulation for severe autism: from pathophysiology to procedure).

[0057] Two major symptom domains that may require treatment in severe autism are social deficits, including nonverbal and unresponsiveness to conversation and potentially life-threatening social impaction (SIB). The amygdala is thought to play a crucial role in the pathophysiology of these abnormalities. Alterations in excitability or inhibitory control are associated with the pathophysiological abnormalities of autism. Neuromodulation targeting the amygdala with deep brain stimulation (DBS) may represent a therapeutic intervention for patients with severe autism. Three cases of DBS treatment have been reported in the literature. The primary objective of the treatment was to alleviate disease-related motor disorders, such as stereotypics (repetitive movement patterns) and self-inflicting 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). The DBS parameters that can be used in the context of the present invention in combination with the nanoparticles described herein are a frequency of 80-130 Hz, a pulse width of 120-210 μs, and a voltage amplitude of 2.5-6.5 V (Sinha et al., Neurosurgery Focus, 2015, 38(6), E3: Deep brain stimulation for severe autism: from pathophysiology to procedure). In one of three cases, DBS in the basal lateral nucleus resulted in a significant improvement in autism-related symptoms, such as social interaction, and was reported to affect modulation and nocturnal sleep (Sturm et al., Frontiers in Human Neuroscience, 2013, 6, 341, 1-10).

[0058] Schizophrenia Schizophrenia is a chronic mental disorder characterized by the following symptoms: positive symptoms reflecting abnormal mental activity (hallucinations and delusions); and negative symptoms corresponding to deficits in normally present mental functions (thought disorders, emotional blunting, and speech impediments). Schizophrenia ranks among the top 10 causes of lifelong disability.

[0059] Significant ventricular enlargement and increased cerebrospinal fluid on the brain surface suggest brain atrophy. This loss of gray matter and 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 the first-onset patient.

[0060] In patients with schizophrenia, observed neural circuit dysfunction has been demonstrated to be due to failure 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).

[0061] Electroconvulsive therapy (ECT), or shock treatment, has been demonstrated 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), and is available herein, in combination with the nanoparticles described herein, for use in the context of the present invention. Electroconvulsive therapy (ECT) involves the continuous application of an electric current to the brain, which induces seizures comparable to epileptic seizures.

[0062] Electrical stimulation for symptomatic treatment of schizophrenia is also possible with DBS. For example, DBS of the nucleus accumbens (NAcc) in depression (frequency 145 Hz, pulse width 90 μs, voltage amplitude 4 V) has resulted in remission of anhedonia, i.e., restoration of pleasure (Schlaepfer et al., Neuropsychopharmacology, 2008, 33, 368-377: Deep brain stimulation to reward circuitry alleviates anhedonia in refractory major depression), and can be used in combination with the nanoparticles described herein in the context of the present invention.

[0063] Examples of cognitive impairment Alzheimer's disease Alzheimer's disease (AD) is a neurodegenerative disorder that leads to progressive decline in mental, behavioral, and functional abilities, as well as learning capacity. Approximately 200,000 people under the age of 65 have AD, making up the younger-onset AD population, while 5 million people are 65 years of age or older.

[0064] Recent evidence suggests that cognitive impairment in Alzheimer's disease is associated with functional disconnection of neural-cognitive networks. Analysis of global EEG synchronization has revealed widespread decreases in alpha, beta, and gamma band synchronization, accompanied by increased delta band synchronization. Loss of beta band synchronization has been shown to correlate with cognitive impairment in patients with mild Alzheimer's disease (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. Typical stimulation parameters usable in the context of the present invention, in combination with the nanoparticles described herein, are a frequency of 130 Hz, a pulse width of 60 or 90 μs, and an amplitude voltage of 3 to 5 V (Laxton et al., World Neurosurgery, 2013, 80, S28.E1-S28.E8: Deep brain stimulation for the treatment of Alzheimer disease and dementias).

[0065] Electrical stimulation In the context of the present invention, the electric field is preferably applied by deep brain stimulation, transcranial electrical stimulation, or transcranial magnetic stimulation. Vagus nerve stimulation (VNS) and spinal cord stimulation can also be applied in the context of the present invention, for example, in the context of epilepsy. Any other known distinct electrical stimulation method, for example, the method described by Grossman N. et al. (Cell, 2017, 169, 1029-1041: Noninvasive deep brain stimulation via temporally interfering electric fields), can be used in the context of the present invention.

[0066] In the context of this invention, the two main brain regions for electrical stimulation are the deep brain and the cerebral cortex.

[0067] Electrical stimulation can reach deep into the brain through surgical implantation of electrodes [the depth of electrode penetration below the skin surface is 10 cm or more, and the penetration range of the electric field generated by the electrodes is several millimeters: deep brain stimulation (DBS)].

[0068] When delivered to the cerebral cortex, electrical stimulation is performed on the surface (the penetration depth of the electric field is usually less than 2 cm below the skin surface; however, according to specific techniques and specific coils for transcranial magnetic stimulation, the electric field can reach a depth of 5 cm). Techniques for providing such an electric field typically include transcranial magnetic stimulation (TMS), repetitive transcranial magnetic stimulation (rTMS), transcranial direct current stimulation (tDCS), high-definition transcranial direct current stimulation (HD-tDCS), transcranial electrical stimulation (TES), transcranial alternating current stimulation (tACS), transcranial pulsed current stimulation (tPCS), and transcranial random noise stimulation (tRNS; alternating current with random amplitude and frequency). The most widely used in clinical trials and preferred in the context of this invention are TMS and tDCS.

[0069] Deep brain stimulation The DBS device includes three key components: stimulating electrodes (also called leads), extension cables, and a programmable pulse generator (PG) similar to a cardiac pacemaker. The device is implanted in two stages. During the first stage, unilateral or bilateral leads are stereotactically implanted in specific therapeutic targets deep within the brain. During the second stage, which may be performed on the same day or later, the pulse generator is implanted under the skin of the anterior chest wall (below the clavicle) or abdomen and connected to the lead wires via subcutaneous tunnel extension cables. The leads are typically 40 cm long, 1.27 mm in diameter, and 1.5 mm or 3 mm wide depending on the indication, providing multiple contact electrodes (most often four contact electrodes, i.e., quadrupole electrodes) spaced 0.5–4 mm apart on the lead. One or two contact electrodes can be stimulated (if two electrodes are used, one is the anode and the other is the cathode). Through the contact electrodes, electrical stimulation is applied directly to deep brain regions, particularly the basal ganglia. Typical currents usable in the context of the present invention are pulsed at high frequencies (100-200 Hz, with 130 Hz being the most frequently used), pulse widths of 60-120 μs, low voltages (less than 4 V), and low currents (less than 2 mA).

[0070] The application of radiofrequency electrical stimulation to the basal ganglia is typically approved (at least in the United States and / or the European Union) and is applicable in the context of the present invention to several movement / motor disorders, such as Parkinson's disease, dystonia, epilepsy, obsessive-compulsive disorder (OCD), and Tourette syndrome.

[0071] Transcranial magnetic stimulation (TMS) Transcranial magnetic stimulation (TMS) is a non-invasive technique used or investigated for numerous research and therapeutic applications, including the study of normal and pathological brain function and the treatment of neurological disorders, and is applicable in the context of this invention. TMS uses short, strong pulses of current delivered to a coil placed in the head of the subject to generate an electric field in the brain via electromagnetic induction. The induced electric field modulates neural activity by modulating the transneuronal potential. The location of activation in the brain is approximately the region where the induced electric field is maximum, and this location is determined by the geometric shape and arrangement of the stimulating coil. Two spatial features of the electric field of interest are the depth of penetration and the focalness, both of which are determined by the geometric shape of the coil and are readily determinable by those skilled in the art. Repetitive TMS (rTMS) is typically used for depression, pain, stroke, etc.

[0072] Transcranial direct current stimulation (tDCS) Transcranial direct current stimulation (tDCS) is a non-invasive technique and is applicable in the context of this invention. In this case, brain stimulation is performed by direct current, resulting in changes in cortical excitability. tDCS is typically performed over a period of 20–35 cm. 2 This method uses a low-intensity (0.5-2mA) constant current applied directly to the head via two electrodes (anode / cathode). Depending on the design, one electrode (reference electrode) can be placed on the forehead (above the supraorbital ridge), and the other electrode (active electrode) can be placed on the contralateral hemisphere, generally on the motor cortex (M1) or dorsolateral prefrontal cortex. The duration of stimulation is most often in the range of 20-40 minutes. A portion of the current penetrates the brain, generating a peak electric field of approximately 0.3 V / m per 1 mA applied. The sustained electric field generated during tDCS can modify transmembrane neuronal potentials, influencing excitability levels and responsiveness to synaptic input, and modulating the firing rate of individual neurons. Increased excitability occurs with anode stimulation, while decreased excitability typically occurs with cathode stimulation.

[0073] tDCS can be used to treat autism (Chi et al., Medical Hypotheses, 2014, 83, 614-618: Treating autism by targeting the temporal lobes), motor rehabilitation after stroke (Gillick et al., Frontiers in Human Neuroscience, 2014, 8(739), 1-9: Pediatric stroke and tDCS: method for rational individualized dose optimization), and major depressive disorder (Croarkin et al., Frontiers in Human Neuroscience, 2014, 8(669), 1-9: Developmental aspects of cortical excitability and inhibition in depressed and healthy youth: an exploratory study).

[0074] nanoparticles This specification describes nanoparticles or nanoparticle aggregates for use in the present invention, for preventing or treating neurological disorders or at least one symptom thereof in a subject when exposed to an electric field. Here, the material of the nanoparticles or nanoparticle aggregates is typically a conductive material, a semiconductor material, or a dielectric constant ε of 200 or more. ijk An insulating material having a dielectric constant ε of 100 or less ijk An insulating material having the following properties is selected.

[0075] composition of nanoparticles Nanoparticles prepared from conductive materials Nanoparticles prepared from conductive materials are either organic or inorganic nanoparticles.

[0076] Inorganic nanoparticles prepared from conductive materials typically exhibit the following characteristics when measured at 25°C and 1 atm pressure relative to a standard hydrogen electrode ("reduction reactions having E° values ​​more positive than that of the standard hydrogen electrode", 8-25, Handbook of Chemistry and Physics; David R. Lide; 88). th (See Table 2 of the Edition) The nanoparticles are prepared from metal elements having a standard reduction potential E° value of about 0.01 or higher, more preferably about 0.1, 0.2, 0.4 or 0.5 or higher. Typical metal elements used to prepare nanoparticles can be selected from Tl, Po, Ag, Pd, Ir, Pt, Au and mixtures thereof. Preferably, the metal elements that can be used as conductive materials for preparing nanoparticles are selected from Ir, Pd, Pt, Au and mixtures thereof.

[0077] Organic nanoparticles prepared from conductive materials are typically prepared from organic materials having adjacent sp2 hybridized carbon centers in their structure (i.e., heteroatoms, typically N or S-containing aromatic rings, either within or outside a carbon double bond or aromatic ring). Preferred organic materials are selected from polyaniline, polypyrrole, polyacetylene, polythiophene, polycarbazole, polypyrene, poly(3,4-ethylenedioxythiophene) and / or poly(3,4-ethylenedioxythiophene)polystyrene sulfonate.

[0078] Nanoparticles prepared from semiconductor materials Nanoparticles prepared from semiconductor materials are typically inorganic nanoparticles.

[0079] Inorganic nanoparticles are typically prepared from semiconductor materials that exhibit a relatively small energy band gap (Eg) between their valence band and conduction band. Typically, semiconductor materials have a band gap Eg of less than 3.0 eV when measured typically at room temperature (25°C). In certain embodiments, the material is either an intrinsic or extrinsic semiconductor material, as further described below herein.

[0080] 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.

[0081] Extrinsic semiconductor materials typically consist of or comprise intrinsic semiconductors prepared to a high degree of chemical purity, where the intrinsic semiconductor material comprises dopants. In certain embodiments, extrinsic semiconductor materials in the form of nanoparticles or nanoparticle aggregates are doped with charge carriers selected from Al, B, Ga, In, and P, if they consist of elements from Group IVA of the Mendeleev periodic table. Such extrinsic semiconductor materials can be either n-type, where negative charge carriers are dominant, or p-type, where positive charge carriers are dominant. 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).

[0082] Nanoparticles prepared from insulating materials with a high relative permittivity, i.e., a relative permittivity of 200 or higher. High relative permittivity ε ijkNanoparticles prepared from or made from insulating materials having a bandgap Eg (also called relative permittivity) typically have a bandgap Eg of 3.0 eV or higher when measured at room temperature (25°C), and at 20°C to 30°C and 10°C. 2 Typical measurements were taken in the Hz to infrared frequency range (e.g., "Permittivity (dielectric constant) of inorganic solid"; Handbook of chemistry and physics; David R. Lide; 88). th (See Tables 12-45 in *Compilation of the static dielectric constant of inorganic solid* by KF Young and HPR Frederikse, J. Phys. Chem. Ref. Data, Vol. 2, No. 2, 1973.) Relative permittivity ε greater than 200 ijk It is prepared from a material having the following characteristics.

[0083] Such nanoparticles are typically prepared from dielectric materials, which are mixed metal oxides preferably selected from BaTiO3, KTaNbO3, KTaO3, SrTiO3, BaSrTiO3, and the like.

[0084] Nanoparticles prepared from insulating materials having a low relative permittivity (relative permittivity), i.e., a relative permittivity of 100 or less. Nanoparticles prepared from or made from insulating materials with low dielectric constant typically have a bandgap Eg of 3.0 eV or higher when measured typically at room temperature (25°C), and exhibit low dielectric constant at 20°C-30°C and 10°C. 2 Typical measurements were taken in the Hz to infrared frequency range (e.g., "Permittivity (dielectric constant) of inorganic solid"; Handbook of chemistry and physics; David R. Lide; 88). th(See Tables 12-45 in "Edition; Compilation of the static dielectric constant of inorganic solid. KF Young and HPR Frederikse. J. Phys. Chem. Ref. Data, Vol. 2, No. 2, 1973") Relative permittivity ε 100 or less, preferably 50 or less or 20 or less. ijk It is prepared from a material having the following characteristics.

[0085] Such nanoparticles are typically prepared from dielectric materials selected from metal oxides, mixed metal oxides, and carbon materials, where the metallic element is a metallic element or lanthanide from periods 3, 5, or 6 of the Mendeleev periodic table. The dielectric material is preferably selected from Al2O3, LaAlO3, La2O3, CeO2, SiO2, SnO2, Ta2O5, ZrO2, HfO2, Y2O3, and carbon diamond.

[0086] Shape of nanoparticles or nanoparticle aggregates The shape of particles or aggregates can affect their "biocompatibility," therefore, particles or aggregates with an extremely homogeneous shape are preferred. For this reason, from a pharmacokinetic standpoint, nanoparticles or aggregates that are essentially spherical, rounded, or oval in shape are preferred. Such shapes are also advantageous for the interaction of nanoparticles or aggregates with cells or for their uptake by cells. Spherical or rounded shapes are particularly preferred.

[0087] The shape of nanoparticles or nanoparticle aggregates is typically evaluated using a transmission electron microscope (TEM).

[0088] Dimensions or size of nanoparticles or nanoparticle aggregates In the spirit of this invention, the terms "nanoparticles" or "nanoparticle aggregates" refer to products having a size in the nanometer range, typically 1 nm to 500 nm, and in particular, synthetic products.

[0089] The term "aggregate of nanoparticles" or "nanoparticles' aggregate" refers to a collection of nanoparticles that are strongly bound together, typically covalently.

[0090] The size of nanoparticles or nanoparticle aggregates can be measured using a transmission electron microscope (TEM). Similarly, the hydrodynamic diameter of nanoparticles or nanoparticle aggregates in solution can be measured using dynamic light scattering (DLS). These two methods can be used sequentially to compare size measurements and confirm the size. The preferred method is DLS (see International Standard ISO22412 Particle Size Analysis - Dynamic Light Scattering, International Organisation for Standardisation (ISO) 2008), while the average hydrodynamic diameter of nanoparticles or nanoparticle aggregates in solution is given by intensity.

[0091] Typically, the maximum dimension or size is the diameter of rounded or spherical nanoparticles or the longest length of oval or elliptical nanoparticles.

[0092] The maximum dimensions of the nanoparticles or aggregates as defined herein are typically about 2 nm to about 250 nm, preferably about 4 nm or 10 nm to about 100 nm or about 200 nm, and more preferably about 10 nm to about 150 nm.

[0093] Biocompatible coatings of nanoparticles or nanoparticle aggregates In a preferred embodiment, 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 agents exhibiting stealth properties may be agents exhibiting stereogroups. Such groups can be selected from, for example, polyacrylates; polyacrylamides (poly(N-isopropylacrylamide)); polycarbamides; biopolymers; polysaccharides, such as dextran or xylan; and collagen. In another preferred embodiment, 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. Agents that form a positive charge on the surface of the nanoparticles or nanoparticle aggregates may be, for example, aminopropyltriethoxysilane or polylysine. The agent that forms a negative charge on the surface of nanoparticles or nanoparticle aggregates may be, for example, a phosphate (e.g., polyphosphate, metaphosphate, pyrophosphate, etc.), a carboxylate (e.g., citrate or dicarboxylic acid, especially succinic acid), or a sulfate.

[0094] In preferred embodiments, nanoparticles or nanoparticle aggregates used in the context of the present invention are coated with a biocompatible material (i.e., a coating agent) selected from hydrophilic agents that present a hydrophilic neutral surface charge or impart a neutral surface charge to the nanoparticles. In practice, when the nanoparticles of the present invention are administered to a subject, nanoparticles coated with a biocompatible agent selected from nanoparticles that present a hydrophilic neutral surface charge or hydrophilic agents that impart a neutral surface charge to the nanoparticles are particularly advantageous in optimizing the use of nanoparticles for treating neurological disorders or at least one symptom thereof when exposed to electrical stimulation / electric fields.

[0095] Hydrophilic agents that impart a neutral surface charge to nanoparticles or nanoparticle aggregates can be agents that present a functional group selected from alcohols (R-OH), aldehydes (R-COH), ketones (R-CO-R), esters (R-COOR), acids (R-COOH), thiols (R-SH), sugars (e.g., glucose, fructose, ribose), acid anhydrides (RCOOOC-R), and pyrrole. Hydrophilic agents that impart a neutral surface charge to nanoparticles or nanoparticle aggregates can be monomers, dimers, oligomers, polymers, or copolymers. If the agent is an oligomer, the oligomer can be an oligosaccharide, such as cyclodextrin. If 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.

[0096] In addition, hydrophilic agents that impart a neutral surface charge to nanoparticles or nanoparticle aggregates can be agents that present a specific group (R-) that can interact with the surface of the nanoparticles or nanoparticle aggregates. R is typically selected from thiols, silanes, carboxylic acids, and phosphate groups.

[0097] When the nanoparticles or nanoparticle aggregates are conductive or semiconductor and metal nanoparticles, 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.

[0098] When the nanoparticles or nanoparticle aggregates are insulators and oxides or mixed oxide nanoparticles, R is preferably a silane or a phosphate group. Preferably, the hydrophilic neutral coating agent is a hydroxymethyltriethoxysilane, fructose 6-phosphate, or glucose 6-phosphate compound.

[0099] The hydrophilic agent that imparts a neutral surface charge to nanoparticles or nanoparticle aggregates can be a zwitterionic compound, such as an amino acid, peptide, polypeptide, vitamin, or phospholipid.

[0100] The surface charge of nanoparticles or nanoparticle aggregates is typically determined, as is well known to those skilled in the art, by zeta potential measurement in water at a pH of 6 to 8 and a nanoparticle concentration of 0.2 to 10 g / L, and by adding an electrolyte at a water concentration of 0.001 to 0.2 M, for example, 0.01 M or 0.15 M. Under the above defined conditions, the surface charge of nanoparticles or nanoparticle aggregates typically falls within the ranges of -10 mV to +10 mV (corresponding to the neutral surface charge), -20 mV to +20 mV, or -35 mV to +35 mV.

[0101] A complete biocompatible coating of 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. "Complete coating" means the presence of a very high density / tightness of biocompatible molecules capable of forming at least a complete monolayer on the surface of the particles.

[0102] Biocompatible coatings enable the stability of nanoparticles, particularly in fluids such as physiological fluids (blood, plasma, serum, etc.) or any isotonic or physiological medium required for drug administration.

[0103] Stability can be confirmed by quantification of the dry extract using a drying oven, and can typically be measured in the nanoparticle suspension before and after filtration through a 0.45 μm filter.

[0104] Advantageously, the coating preserves the integrity of the particles in vivo, ensures or improves their biocompatibility, and facilitates any functionalization (e.g., by spacer molecules, biocompatible polymers, targeting agents, proteins, etc.).

[0105] The biocompatible nanoparticles or nanoparticle aggregates of the present invention should not dissolve and release toxic species after in vivo administration (i.e., at physiological pH), nor should they exhibit redox behavior in the absence of electrical stimulation.

[0106] Another specific object described herein is, preferably, a composition comprising the nanoparticles and / or nanoparticle aggregates defined above, together with a pharmaceutically acceptable carrier or medium, in particular a pharmaceutical composition.

[0107] In particular, this specification describes compositions for use in preventing or treating neurological disorders or at least one symptom thereof as described herein in an object exposed to an electric field. Herein, the composition comprises or consists of nanoparticles and / or nanoparticle aggregates and a pharmaceutically acceptable support, wherein the material of the nanoparticles or nanoparticle aggregates is typically a conductive material, a semiconductor material, or a dielectric constant ε of 200 or more, as described herein. ijk An insulating material having a dielectric constant ε of 100 or less ijk An insulating material having the following properties is selected.

[0108] In a preferred embodiment, the composition comprises or consists of at least two distinct nanoparticles and / or nanoparticle aggregates, each nanoparticle or nanoparticle aggregate made of a distinct material, typically a conductive material, a semiconductor material, and a dielectric constant ε of 200 or more. ijk An insulating material having a dielectric constant ε of 100 or less ijk An insulating material having the following properties is selected.

[0109] In certain embodiments, the composition may include, together with the therapeutic agent, nanoparticles or nanoparticle aggregates of the present invention. The therapeutic agent can be selected from any drug used to treat neurological disorders. The therapeutic agent is typically selected from antipsychotics, anti-dopaminergic agents, dopaminergic agents, anticholinergic agents, cholinergic agents, anti-glutamate agents, glutamate agents, acetylcholinesterase inhibitors, N-methyl D-aspartate (NMDA) receptor antagonists, gamma-aminobutyric acid (GABA) agonists, botulinum toxin, anti-dystonia agents, antiepileptic agents, anticonvulsants, mood stabilizers, antidepressants, and sedatives.

[0110] The composition may be in the form of a solid, liquid (particles in a suspension), aerosol, gel, paste, etc. A preferred composition is in the form of a liquid or gel. A particularly preferred composition is in the form of a liquid.

[0111] The pharmaceutically acceptable support or carrier used may be any classic support that is relevant to those skilled in the art, such as physiological saline, isotonic, sterile, buffered solutions, non-aqueous media solutions, etc.

[0112] Furthermore, the composition may also contain stabilizers, sweeteners, surfactants, polymers, and the like.

[0113] The composition can be formulated, for example, as an ampoule, aerosol, bottle, tablet, or capsule using pharmaceutical formulation techniques known to those skilled in the art.

[0114] The nanoparticles or nanoparticle aggregates of the present invention can be administered to a subject using various possible routes, such as intracranial, intravenous (IV), airway (inhalation), intrathecal, intraocular, or oral (oral) routes, preferably intracranial or intrathecal.

[0115] If necessary, the nanoparticles can be repeatedly injected or administered.

[0116] The nanoparticles or nanoparticle aggregates described herein and compositions comprising such nanoparticles or nanoparticle aggregates are intended for use in subjects, typically in animals, preferably in mammals, and more preferably in humans, typically in human patients, regardless of age or sex.

[0117] The typical amount of nanoparticles or nanoparticle aggregates administered to the target cerebral cortex is 10 5 ~10 15 pieces, preferably 10 7 ~10 14 Individual, comfortable, 10 9 ~10 12 The number is [number]. Also, the typical amount of nanoparticles or nanoparticle aggregates administered to the target cerebral cortex is 1 cm³. 3 10 2 ~10 12 It consists of individual nanoparticles or nanoparticle aggregates.

[0118] The typical amount of nanoparticles or nanoparticle aggregates administered to the deep brain of the target is 10 4 ~10 14 pieces, preferably 10 6 ~10 12 Individual, comfortable, 10 8 ~10 11 The number is [number]. Furthermore, the typical amount of nanoparticles or nanoparticle aggregates administered to the deep brain of the target is 1 cm³. 3 10 1 ~10 11 It consists of individual nanoparticles or nanoparticle aggregates.

[0119] In the context of the present invention, exposing nanoparticles or nanoparticle aggregates to an electric field / electrical stimulation is equivalent to exposing a subject to which nanoparticles or nanoparticle aggregates have been administered to an electric field / electrical stimulation.

[0120] Furthermore, this specification describes a method for preventing or treating a neurological disorder or at least one symptom thereof in a subject, comprising the steps of administering one of the nanoparticles or nanoparticle aggregates described herein to the subject, and exposing the subject to an electric field / electrical stimulation.

[0121] Further object described herein is to include at least two distinct nanoparticles and / or at least two distinct nanoparticle aggregates described herein, each nanoparticle or nanoparticle aggregate made of a distinct material is typically a conductive material, a semiconductor material, or a dielectric constant ε of 200 or more as described herein. ijk An insulating material having a dielectric constant ε of 100 or less ijk The kit relates to an insulating material having [specific properties].

[0122] In certain embodiments, the kit includes, in a separate container, separate nanoparticles and / or nanoparticle aggregates described herein (these are intended to be mixed either in vitro or ex vivo, typically in contact with, i.e., at, the target site, or before the deposition of the mixture at the target site).

[0123] A further object is a kit further comprising, unlike the nanoparticles or nanoparticle aggregates described herein, at least one further therapeutic agent, such as an antipsychotic, anti-dopamine agonist, dopamine agonist, anticholinergic agonist, cholinergic agonist, anti-glutamate agonist, glutamate agonist, acetylcholinesterase inhibitor, N-methyl D-aspartate (NMDA) receptor antagonist, gamma-aminobutyric acid (GABA) agonist, botulinum toxin, anti-dystonia agent, antiepileptic agent, anticonvulsant, mood stabilizer, antidepressant, and sedative.

[0124] Furthermore, this specification describes the in vivo, in vitro, or ex vivo use of such kits in methods for preventing or treating neurological disorders or at least one symptom thereof described herein in subjects. Also, this specification discloses kits described herein for use in the prevention or treatment of neurological disorders or at least one symptom thereof in subjects.

[0125] The present invention aims to treat neurological disorders or at least one symptom thereof by using nanoparticles or nanoparticle aggregates exposed to electrical stimulation / electric fields.

[0126] At the neuronal level, nanoparticles have been described as either enhancing or inhibiting 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).

[0127] A systemic effects study 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)] in neuronal systems demonstrated that the nanoparticles induced changes in mechanisms affecting excitability. Furthermore, neuronal network simulations showed that locally cAgNP-induced changes led to changes in the network activity of the entire network, indicating that local application of cAgNPs can influence 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).

[0128] Furthermore, the increased neuronal excitability associated with intracellular gold nanoparticles has been reported to potentially have adverse effects on neurons in pathological conditions, such as during 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).

[0129] The nanoparticles or nanoparticle aggregates of the present invention are intended for use in preventing or treating neurological disorders or at least one symptom thereof by normalizing the synchronization of oscillations within and / or between neuronal networks in and / or between neuronal networks in and between distinct regions of the brain when exposed to an electric field / electrical stimulation.

[0130] As illustrated in Figures 2 and 3, transmission within and / or between distinct regions of the brain is affected in neurological disorders. According to neurological disorders and associated symptoms, exposure of specific regions of the brain to the nanoparticles of the present invention (see Table 2), when combined with electrical stimulation, will improve transmission by normalizing the synchronization of oscillations within and / or between neural networks within and between distinct regions of the brain (i.e., normalization of coherence) (Figures 4 and 5 and Table 2).

[0131] [Table 2]

[0132] As those skilled in the art will readily understand, the effect of electrical stimulation on neural networks is related to the penetration depth and spatial resolution of the electric field within the target brain region. Low spatial resolution and penetration depth are significant drawbacks of electrical stimulation. The presence of nanoparticles or nanoparticle aggregates of the present invention advantageously enables improved spatial resolution (focus) of the electric field to which the nanoparticles are localized and improved penetration depth of the current (improvement of its therapeutic effect).

[0133] Furthermore, the presence of nanoparticles or nanoparticle aggregates in the target tissue can reduce the applied / induced electrical stimulation threshold required for neuronal stimulation, i.e., it reduces the values ​​of applied parameters such as current, voltage, pulse width, and / or frequency. In addition, this effect reduces the potential toxicity associated with applied / induced current. This can also have technical implications such as extending the lifespan of the internal pulse generator (IPG) battery or modifying (reducing) the size and geometric shape of the DBS electrode.

[0134] The following embodiments and their corresponding drawings illustrate the present invention without limiting its scope. [Brief explanation of the drawing]

[0135] [Figure 1] Schematic diagram of the brain (sagittal plane). [Figure 2] Oversynchronization and synchronization failure between two neuronal networks. [Figure 3] Brain regions involved in various neurological diseases. [Figure 4] The effect of normalizing hypersynchronization (motion disorder) when nanoparticles (NP1) are exposed to an electric field (E). [Figure 5] The effect of nanoparticles (NP2) being exposed to an electric field (E) on normalizing synchronous disorders (mental and cognitive impairments). [Figure 6] Experimental scheme for Parkinson's disease induction with MPP+ treatment and electrical activity recording. Co-cultures of ventral midbrain / cortex from mice were prepared from E14.5 NMRI mice and cultured in 48 wells of MEA for 3 weeks (culture period). On day 7 after culturing, these cultures were treated with a nanoparticle suspension ("nanoparticle" group) or water ("control" group and "MPP+" group), and on day 8, they were treated with MPP+ (20 μM) ("nanoparticle" group and "MPP+" group) or water ("control" group). Spontaneous activity was recorded on day 21. After recording on day 21, the cultures were electrically stimulated on one electrode, and activity was recorded on the non-stimulated electrode. [Figure 7]Two simplified burst schemes outline several parameters that can be extracted from electrical activity recordings. Parameters describing overall activity (spikes, bursts, inter-burst interval (IBI), and burst duration) and burst structure (burst duration, burst plateau, burst amplitude, burst inter-spike interval (ISI), and burst area) are shown. The standard deviations (SD) of these parameters are measures of the regularity of the overall activity and burst structure, respectively. The coefficient of variation over time (CVtime) reflects the temporal regularity of the activity pattern in each unit. 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 a network. CVnet is calculated by the ratio of the parameter's standard deviation to its mean across the network. A large CVnet value indicates widespread variability in activity across the entire network and less synchronization. [Figure 8] The functional effects observed in the "nanoparticle" group under electrical stimulation were compared with the "control" group (under electrical stimulation) and the "MPP+" group (under electrical stimulation) for mesobrain / cortical network activity. All MPP+-induced functional effects in network activity under electrical stimulation, both in and out of the tested nanoparticles, as well as the "control" group (under electrical stimulation), were normalized to "pre-stimulation" activity, i.e., activity measured on day 21, set to 100% for each experiment. The data demonstrate the MPP+-induced functional effects under electrical stimulation, and the preventive / rescue efficacy of the nanoparticles of the present invention under electrical stimulation (i.e., the ability to prevent / rescue functional effects to a level similar to that of the "control" group). [Figure 9] Effectiveness score analysis for the "nanoparticle" group, the "control" group (effectiveness score = 0), and the "MPP+" group (effectiveness score = 1). [Figure 10]An experimental scheme for inducing, treating, and recording electrical activity of Alzheimer's disease with amyloid beta 1-42 (A-beta 1-42). After 4 weeks of culture (culture period), A-beta 1-42 (100 nM) ("nanoparticle" group and "A-beta" group) or water ("control" group) (T0) was added to the neuronal network. After 4 hours, a nanoparticle suspension ("nanoparticle" group) or water ("control" group and "A-beta" group) was added. Spontaneous activity was recorded as follows: - at T0 (before addition of A-beta 1-42), - at T0+1h, T0+2h, T0+3h, T0+4h (before addition of nanoparticles or water), T0+5h, and T0+6h. [Figure 11] The functional effects observed in the "nanoparticle" group under electrical stimulation were compared with those of the "control" group (under electrical stimulation) and the "A-beta 1-42" group (under electrical stimulation) for cortical network activity. All A-beta 1-42-induced functional effects in network activity under electrical stimulation, both in and out of the tested nanoparticles, as well as the "control" group (under electrical stimulation), were normalized to "pre-stimulation" activity, i.e., activity measured at T0+6 time, set to 100% for each experiment. The data demonstrate the functional effects of A-beta 1-42 under electrical stimulation and the rescue efficacy (i.e., the ability to rescue functional effects to a level similar to that of the "control" group) made possible by the nanoparticles of the present invention under electrical stimulation. [Figure 12] Effectiveness score analysis for the "nanoparticle" group, the "control" group (effectiveness score = 0), and the "A-beta" group (effectiveness score = 1). [Examples]

[0136] simulation Simulations can be used to evaluate the effects of nanoparticles exposed to electrical stimulation (electric fields) on neuronal networks.

[0137] In vitro research on neurons At the neuronal level, patch-clamp techniques are extremely useful for detecting action potentials because they allow for the simultaneous direct measurement and control of the neuronal membrane potential.

[0138] This technology will be used to evaluate the effects of nanoparticles on a single neuron.

[0139] In vitro research on neural networks Multi-electrode arrays (MEAs) enable the stimulation and recording of a large number of neurons (neuronal networks). Isolated neuronal cultures on an MEA provide a simplified model in which network activity can be manipulated by electrical stimulation sequences through multiple electrodes on the array. This technique is extremely useful for evaluating physiologically related issues at the network and cellular levels, leading to a better understanding of brain function and dysfunction.

[0140] In fact, isolated neuronal cultures bound to MEA are widely used to better understand the complexity of brain networks. In addition, the use of isolated neuronal assemblies allows for the manipulation and control of network connectivity. The use of isolated neuronal cultures bound to MEA enables the design of experiments in which neurons can be stimulated extracellularly by electrical pulses delivered through the same electrodes of the apparatus. Thus, it becomes reasonable to investigate how emerging neuronal dynamics can be modulated by electrical stimulation and, as a result, whether underlying functional connectivity is modified (Poli D. et al, Frontiers in Neural Circuits, 2015, 9 (article 57), 1-14: Functional connectivity in in vitro neuronal assemblies).

[0141] The MEA system enables non-invasive, long-lasting, simultaneous extracellular recording from multiple sites in a neuronal network in real time, providing robust measurements of network activity through improved spatial resolution. Simultaneous acquisition of action potential and electric field potential data over extended periods allows for monitoring of network function arising from the interactions of all cellular mechanisms responsible for spatiotemporal pattern generation (Johnstone AFM et al., Neurotoxicology (2010), 31: 331-350, Microelectrode arrays: a physicologically based neurotoxicity testing platform for the 21 stCompared to patch-clamp and other single-electrode recording techniques, MEA measures the response of the entire network and integrates comprehensive information about the interactions of all receptors, synapses, and neuron types present in the network (Novellino A. et al., Frontiers in Neuroengineering. (2011), 4(4), 1-14, DeVelopment of micro-electrode array based tests for neurotoxicity: assessment of interlaboratory reproducibility with neuroactive chemicals.). Therefore, MEA recordings have been used to understand neuronal transmission, information encoding, propagation, and processing in neuronal culture (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). MEA technology is an advanced phenotypic high-content screening method for characterizing functional changes in network activity in electrically active cell cultures, and is highly sensitive to neurogenesis, neurogenesis, and neurodegeneration.Furthermore, it is known that neuronal networks grown on MEAs can respond to neuroactive or neurotoxic compounds at almost the same concentration ranges that alter the function of intact mammalian nervous systems (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 150MHz carrier wave pulsed with an alternating 10 and 16 Hz modulation).

[0142] This technology will be used to evaluate the effects of nanoparticles on neuronal networks.

[0143] In vivo research on neuronal networks To evaluate the effects of the nanoparticles of the present invention on animal neuronal networks when exposed to electrical stimulation, appropriate animal models should be considered.

[0144] For example, we will use a mouse model of Parkinson's disease to evaluate the effect of nanoparticles stimulated by tDCS (transcranial direct current stimulation) on reducing behavioral impairments (motor impairments). We will also use a rat model of Alzheimer's disease to evaluate the effect of nanoparticles stimulated by tDCS on spatial learning and memory dysfunction (cognitive impairment) in animals.

[0145] Example 1. Nanoparticles prepared from conductive material: Synthesis of gold nanoparticles coated with a biocompatible coating having a neutral surface charge. Gold nanoparticles were synthesized by reducing gold chloride salt (HAuCl4) with a capping agent (sodium citrate) (the protocol was adopted from G. Frens Nature Physical Science 241 (1973) 21). In a typical experiment, the HAuCl4 solution was heated to a boil. Subsequently, a sodium citrate solution was added. The resulting solution was maintained under boiling conditions for a further 5 minutes. The nanoparticle suspension was filtered through a 0.22 μm filter (filter membrane: poly(ethersulfone) (PES)), and the gold concentration in the suspension was determined by UV-Vis spectroscopic analysis assay at 530 nm.

[0146] Surface coating was performed using α-methoxy-ω-mercaptopoly(ethylene glycol) 20kDa ("Thiol-PEG 20kDa"). A sufficient amount of "Thiol-PEG 20kDa" was added to the nanoparticle suspension to reach at least half of the monolayer coating on the surface of the gold nanoparticles (2.5 molecules / nm). 2 The pH was adjusted to 7-7.2, and the nanoparticle suspension was stirred overnight.

[0147] The hydrodynamic diameter (measured by intensity) was determined by dynamic light scattering (DLS) using a Nano-Zetasizer (Malvern) with a 633 nm laser at a scattering angle of 173° after diluting the nanoparticle suspension in water (final concentration: 0.1 g / L). The hydrodynamic diameter of the biocompatible gold nanoparticles obtained in this way in the suspension was found to be equal to 118 nm, and the polydispersity index (dispersion of the nanoparticle population in terms of size) was found to be 0.13.

[0148] The zeta potential was determined by measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern) after diluting the nanoparticle suspension in a 1 mM NaCl solution at pH 7 (final concentration: 0.1 g / L). The zeta potential at pH 7 was found to be equal to -1 mV.

[0149] Example 2. Nanoparticles prepared from conductive material: Synthesis of gold nanoparticles coated with a biocompatible coating having a negative surface charge. Gold nanoparticles were prepared as described in Example 1 (same gold inorganic core).

[0150] The gold concentration in the suspension was determined by 0.22 μm filtration using a PES membrane filter and UV-Vis spectroscopic analysis assay at 530 nm.

[0151] Biocompatible surface coating was performed using meso-2,3-dimercaptosuccinate (DMSA). A sufficient amount of DMSA was added to the nanoparticle suspension to reach at least half of the monolayer coating on the surface (2.5 molecules / nm). 2 The pH was adjusted to 7-7.2, and the nanoparticle suspension was stirred overnight.

[0152] The hydrodynamic diameter (measured by intensity) was determined by dynamic light scattering (DLS) using a Nano-Zetasizer (Malvern) with a 633 nm laser at a scattering angle of 173° after diluting the nanoparticle suspension in water (final concentration: 0.1 g / L). The hydrodynamic diameter of the nanoparticles obtained in this way in the suspension was equal to 76 nm, and the polydispersity index (dispersion of the nanoparticle population in terms of size) was 0.46.

[0153] The zeta potential was determined by measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern) after diluting the nanoparticle suspension in a 1 mM NaCl solution at pH 7 (final concentration: 0.1 g / L). The zeta potential at pH 7 was found to be equal to -23 mV.

[0154] Example 3. Nanoparticles prepared from insulating materials with a low dielectric constant of 100 or less: Synthesis of zirconium oxide nanoparticles coated with a biocompatible coating having a neutral surface charge. Zirconium oxide (ZrO2) nanoparticles were synthesized by precipitating zirconium chloride (ZrCl4) with tetramethylammonium hydroxide (TMAOH) at a basic pH. The resulting suspension was transferred to an autoclave and heated to a temperature exceeding 110°C. After cooling, the suspension was washed with deionized water to make it acidic.

[0155] The concentration of (ZrO2) nanoparticles was determined by filtration at 0.22 μm using a PES membrane filter, drying the aqueous solution into a powder, and weighing the resulting mass.

[0156] A biocompatible coating was prepared using silane-poly(ethylene) glycol 2kDa ("Si-PEG 2kDa"). A sufficient amount of "Si-PEG 2kDa" was added to the nanoparticle suspension to reach at least half of the monolayer coating on the surface (2.5 molecules / nm). 2 The nanoparticle suspension was stirred overnight, and then the pH was adjusted to 7.

[0157] The hydrodynamic diameter (measured by intensity) was determined by dynamic light scattering (DLS) using a Nano-Zetasizer (Malvern) with a 633 nm laser at a scattering angle of 173° after diluting the nanoparticle suspension in water (final concentration: 0.1 g / L). The hydrodynamic diameter of the nanoparticles was found to be equal to 55 nm, and the polydispersity index (dispersion of the nanoparticle population in terms of size) was found to be 0.1.

[0158] The zeta potential was determined by measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern) after diluting the nanoparticle suspension in a 1 mM NaCl solution at pH 7 (final concentration: 0.1 g / L). The zeta potential at pH 7 was found to be equal to -1 mV.

[0159] Example 4. Nanoparticles prepared from insulating materials with a low dielectric constant of 100 or less: Synthesis of zirconium oxide nanoparticles coated with a biocompatible coating having a negative surface charge. Zirconium oxide nanoparticles were prepared as described in Example 3 (same inorganic core).

[0160] The concentration of (ZrO2) nanoparticles was determined by filtration at 0.22 μm using a PES membrane filter, drying the aqueous suspension into a powder, and weighing the resulting mass.

[0161] Surface functionalization was performed using sodium hexametaphosphate. A sufficient mass of sodium hexametaphosphate was added to the nanoparticle suspension to reach at least half of the monolayer coating on the surface (2.5 molecules / nm). 2 The nanoparticle suspension was stirred overnight, and then the pH was adjusted to 7.

[0162] The hydrodynamic diameter (measured by intensity) was determined by dynamic light scattering (DLS) using a Nano-Zetasizer (Malvern) with a 633 nm laser at a scattering angle of 173° after diluting the nanoparticle suspension in water (final concentration: 0.1 g / L). The hydrodynamic diameter of the nanoparticles was found to be equal to 70 nm, and the polydispersity index (dispersion of the nanoparticle population in terms of size) was found to be 0.11.

[0163] The zeta potential was determined by measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern) after diluting the nanoparticle suspension in a 1 mM NaCl solution at pH 7 (final concentration: 0.1 g / L). The zeta potential at pH 7 was found to be equal to -33 mV.

[0164] Example 5. Nanoparticles prepared from semiconductor materials: Silicon nanoparticles coated with a biocompatible coating having a negative surface charge. Silicon (Si) nanoparticles (powder) were obtained from US Research Nanomaterials Inc. They were dispersed in water at a concentration of 30 g / L under sonication (using a probe).

[0165] The concentration of (Si) nanoparticles was determined by filtration at 0.22 μm using a PES membrane filter, drying the suspension into a powder, and weighing the resulting mass.

[0166] The hydrodynamic diameter (measured by intensity) was determined by dynamic light scattering (DLS) using a Nano-Zetasizer (Malvern) with a 633 nm laser at a scattering angle of 173° after diluting the nanoparticle suspension in water (final concentration: 0.1 g / L). The hydrodynamic diameter of the nanoparticles was found to be equal to 164 nm, and the polydispersity index (dispersion of the nanoparticle population in terms of size) was found to be 0.16.

[0167] The zeta potential was determined by measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern) after diluting the nanoparticle suspension in a 1 mM NaCl solution at pH 7 (final concentration: 0.1 g / L). The zeta potential at pH 7 was found to be equal to -19 mV.

[0168] Example 6. Nanoparticles prepared from an insulating material having a high dielectric constant 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).

[0169] Surface functionalization was performed using silane-poly(ethylene) glycol 10kDa ("Si-PEG 10kDa"). Briefly, "Si-PEG 10kDa" was first dissolved in an ethanol / water solution (1 / 3 v / v) and added to a BaTiO3 suspension (20 wt% in water) to achieve a complete monolayer coating on the surface of the nanoparticles. This suspension was sonicated and then stirred overnight. After 0.22 μm filtration (filter membrane: poly(ethersulfone)), a washing step was performed to remove unreacted "Si-PEG 10kDa" polymer.

[0170] The hydrodynamic diameter (measured by intensity) was determined by dynamic light scattering (DLS) using a Nano-Zetasizer (Malvern) with a 633 nm laser at a scattering angle of 173° after diluting the nanoparticle suspension in water (final concentration: 0.1 g / L). The hydrodynamic diameter of the nanoparticles was found to be equal to 164 nm, and the polydispersity index (dispersion of the nanoparticle population in terms of size) was found to be 0.16.

[0171] The zeta potential was determined by measuring the electrophoretic mobility of the nanoparticles (Nano-Zetasizer, Malvern) after diluting the nanoparticle suspension in a 1 mM NaCl solution at pH 7 (final concentration: 0.1 g / L). The zeta potential at pH 7 was found to be -11 mV.

[0172] Example 7. MPP using phenotypic MEA screening technology + Evaluation of the preventive / rescue efficacy of nanoparticles from Examples 1, 2, 5, and 6 exposed to electrical stimulation in evoked neuronal networks. The preventive / rescue efficacy of the nanoparticles of the present invention was demonstrated by culturing them on 48-well MEA for 3 weeks in MPP + The compounds were tested in co-cultures of ventral midbrain / cortex from treated mice. This model represents an in vitro Parkinsonian model for screening compounds based on the functional salvation of dopaminergic neurons using cultures of impaired midbrain / cortex growing on MEAs. The midbrain is a region of the brain that is part of the basal ganglia and includes the substantia nigra, which contains the majority of dopaminergic neurons. The preventive / salvage effects of nanoparticles were evaluated by measuring the extracellular electrical activity of co-cultures of neurons seeded on microelectrode array (MEA) chips.

[0173] In vitro induction of the Parkinsonian phenotype in mouse neurons using 1-methyl-4-phenylpyridinium iodide (MPP) + This was done by [method / organization name]. There is strong evidence that mitochondrial dysfunction plays a role in the pathogenesis of Parkinson's disease (PD). MPP +It was found to be a mitochondrial toxin that inhibits cellular respiration by blocking electron transport enzyme complex I (NADH: ubiquinone oxidoreductase). Several laboratories have reported the presence of selective deficiency of mitochondrial electron transport chain complex I in the substantia nigra of postmortem tissue from PD patients, and decreased complex I activity in platelets from early-stage PD patients (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.).

[0174] Materials and methods Primary cell culture, processing conditions, and electrical stimulation Midbrain tissue was collected from 14.5-day-old embryonic chr:NMRI mice (Charles River). Mice were euthanized by cervical vertebral dislocation. The tissue was separated by enzymatic digestion (133,3 kunitz units / ml DNase; 10 units / ml Papain) and mechanical grinding, counted, and viability controlled. It was then seeded on MEA in 20 μl drops of DMEM containing laminin (10 μg / ml), 10% fetal bovine serum, and 10% equine serum. The cultures on the MEA were incubated at 37°C in a 10% CO2 atmosphere until ready for use. DMEM containing 10% equine serum was replenished in the culture medium twice a week.

[0175] In the "nanoparticle" group, on day 7, the wells were filled with nanoparticle suspensions from Examples 1, 2, and 5 (800 μM) and from Example 6 (2000 μM), and then on day 8, 20 μM MPP was used. + The treatment was performed using the following method. In the "control" group, water was added to the wells on day 7, and then again on day 8. + In the group, water was added to the wells on day 7, followed by 20 μM MPP on day 8. +Added MPP + After 24 hours (or 24 hours) of addition (or water for the "control" group), the medium is replaced and MPP is applied. + The cleaning was completed. After that, the culture medium was changed twice a week.

[0176] On day 21, neuronal activity was recorded for 120 minutes, and stable activity for 30 minutes was analyzed. After recording on day 21, all wells were activated with one electrode that actively spiked them with electrical stimulation. Stimulation was performed for 30 minutes (one electrode stimulation per well in a 48-well MEA, minimum stimulation duration = 100 μs, artifact removal 2 ms after pulse, pulse 10 × biphasic + / - 500 mV). The response of the unstimulated electrode was averaged and normalized to pre-stimulation activity (Figure 6).

[0177] Microelectrode array neurochip We purchased 48-well microelectrode array neurochips from Axion Biosystems Inc. These chips have 16 passive electrodes per well. The surface was coated with polyethyleneimine (PEI, 50% in borate buffer) for 1 hour, washed, and air-dried.

[0178] Multichannel recording and multiparametric data analysis A multi-channel MAESTRO recording system from Axion Biosystems (USA) was used for recording. For extracellular recording, 48 wells of MEA were placed in the MAESTRO recording station and maintained at 37°C. Recording was performed in DMEM / 10% thermoinactivated horse serum. pH was maintained at 7.4 by a continuous flow of filtered, humidified air containing 10% CO2.

[0179] Each unit represents activity originating from a single neuron recorded by a single electrode. Units are separated at the start of recording. For each unit, action potentials (i.e., spikes) are recorded as spike trains and clustered into so-called "bursts." 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 start and end of short spike events (Figure 7).

[0180] Multiparametric high-content analysis of network activity patterns extracted 204 activity-descriptive spike column parameters. These parameters allow for accurate descriptions of activity changes in the following four categories: overall activity, burst structure, oscillation behavior, and synchronous behavior. - Changes in "overall activity parameters" describe the effects on action potential firing rate (spike rate), burst rate, and burst duration as the time between bursts. - The "burst structure parameters" define not only the internal structure of spikes within a high-frequency spike phase ("burst"), such as the spike frequency, spike velocity, and burst spike density within the burst, but also the overall structure of the burst, such as its duration, area, and plateau. - The "oscillating parameter" quantifies the regularity of burst occurrence or structure, and this regularity is calculated by the coefficient of variation of the main activity parameter that describes the variability of the parameter (overall activity, burst structure) within the experimental episode (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 less regular burst structure or less regular overall activity (e.g., spikes, bursts). - As a measure of synchronism in spike trains, the "CVnet parameter" reflects the "synchronization" between neurons in the 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 large CVnet value indicates widespread variation in activity across the entire 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).

[0181] MPP in neuronal networks under electrical stimulation + The functional effects induced by and the preventive / relief efficacy of the nanoparticles of the present invention under electrical stimulation were evaluated through the above parameters (some of which are summarized in Table 3 below).

[0182] [Table 3]

[0183] MPP in network activity under electrical stimulation in the presence or absence of tested nanoparticles + Evoked functional effects were normalized to “pre-stimulation” activity, i.e., activity measured on day 21, set to 100% for each experiment. Values ​​related to spontaneous native activity were obtained from 60-second bin data taken from the 30-minute period 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, and for the “control” group, at least 30 active wells were used. + For the "active" group, at least 26 active wells ("active" meaning wells with a sufficient number of electrodes to measure electrical activity) were included in the analysis. The absolute parameter distribution was tested for normality, and statistical significance between groups was assessed by one-way ANOVA.

[0184] Figure 8 shows some representative parameters from the following categories: vibration behavior and synchronization. These parameters determine MPP under electrical stimulation. + The invention is characterized by its induced functional effects and the preventive / relief efficacy enabled by the nanoparticles of the present invention under electrical stimulation (i.e., the ability to prevent / relieve functional effects to a level similar to that of the "control" group).

[0185] To evaluate the effect of the compound, the multiparametric results of selecting 204 parameters were expressed as a single parameter called the "effect score." This is a linear combination of the selected features, and the dataset was divided into a "control" group exposed to the electric field with a mean value of "0" and a "MPP" group exposed to the electric field with a mean value of "1." + Convert to a vector containing the "control" group and the "MPP + The best distinction between the two groups was determined through the selection of 18 features out of 204 parameters that were measured and optimized (Kummel A, et al. J Biomol Screen., 2010, 15(1),95-101: Integration of multiple readouts into the z' factor for assay quality assessment). The effect score analysis is shown in Figure 9.

[0186] Table 4 shows the preventive / relief efficacy of the nanoparticles of the present invention when exposed to electrical stimulation.

[0187] [Table 4]

[0188] The treatment of Parkinson's disease symptoms with DBS has been approved by the FDA since 2002. In combination with the nanoparticles described herein, the most commonly used stimulation parameters available in the context of this invention are a frequency of 130–185 Hz, a pulse width of 60–210 μs, and a voltage amplitude of 1–3.5 V. In the experiments described herein, stimulation was applied to a neuronal network coculture for 30 minutes. Stimulation = 10 biphase pulses (pulse duration = 100 μs), pulse amplitude = + / - 500 mV, pulse frequency = 20 Hz, and pulse train duration = 0.2 Hz.

[0189] Figures 12, 13 and Table 4 show that pretreatment of a neuronal network with the nanoparticles of the present invention and exposure to an electric field resulted in MPP in the neuronal network under an electric field. +The induced functional effects demonstrate prevention / relief. Interestingly, the preventive / relief efficacy can be observed for parameters in categories related to oscillatory behavior and synchronization, reaching levels observed in the “control” group. These oscillatory behavior and synchronization parameters are typically monitored as measures of altered network development. These parameters can be advantageously relieved in the presence of the nanoparticles of the present invention, which are exposed to electrical stimulation.

[0190] These results suggest that when exposed to an electric field, MPP in neuronal networks occurs under electric field conditions. + The advantageous performance of the nanoparticles described in this application in rescuing induced functional effects is highlighted.

[0191] Example 8. Evaluation of the effects of nanoparticles from Examples 2, 3, 4, and 5 exposed to electrical stimulation 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, exposed to electrical stimulation, was tested in vitro via MEA in an amyloid-beta 1-42 (A-beta 1-42) induced model of Alzheimer's disease in frontal cortical cultures of mouse neurons.

[0192] To induce Alzheimer's-related functional phenotypes, synthetic HFIP (hexafluoroisopropanol)-treated A-beta 1-42 peptide (HFIP treatment generates amyloid-beta monomers) is used at a subtoxic dose (100 nM). High levels of amyloid-beta (A-beta) reduce glutamatergic synaptic transmission and cause synaptic loss (Palop et al., Nat Neurosci., 2010, 13(7), 812-818: Amyloid-beta induced neuronal dysfunction in Alzheimer's disease: from synapses toward neural networks; Hsia et al., Proc.Natl. Acad. Sci., 1999, 96, 3228-3233: Plaque-independent disruption of neural circuits in Alzheimer's disease mouse models). The production of A-beta and its secretion into the extracellular space are strictly regulated by neuronal activity in vitro and in vivo. Increased neuronal activity enhances A-beta production, while blocking neuronal activity has the opposite effect. This synaptic regulation of A-beta is mediated, at least partially, by clathrin-dependent endocytosis of surface amyloid precursor protein (APP) at the presynaptic terminal, endosomal proteolytic cleavage of APP, and A-beta release at the synaptic terminal (Cirrito et al., Neuron, 2005, 48, 913-922: Synaptic activity regulates interstitial fluid amyloid-beta levels in vivo).

[0193] Materials and methods primary cell culture Frontal cortical tissue was collected from embryonic day 15 / 16 chr:NMRI mice (Charles River). Mice were euthanized by cervical vertebral dislocation. The tissue was separated by enzymatic digestion (133,3 kunitz units / ml DNase; 10 units / ml Papain) and mechanical grinding, counted, and viability controlled. It was then seeded on MEA in 20 μl drops of DMEM containing laminin (10 μg / ml), 10% fetal bovine serum, and 10% equine serum. The cultures on the MEA were incubated at 37°C in a 10% CO2 atmosphere until ready for use. DMEM containing 10% equine serum was replenished in the culture medium twice a week. On day 5 after seeding, the developing co-cultures were treated with the mitotic inhibitors 5-fluoro-2'-deoxyuridine (25 μM) and uridine (63 μM) to prevent further glial proliferation.

[0194] In the "nanoparticle" group, first, the wells were treated with A-beta 1-42 (synthetic HFIP-treated amyloid-beta 1-42 peptide) at T0 (T0 being the end of the 28-day in vitro culture period). Then, the wells were treated with nanoparticle suspensions from Examples 2, 3, 4, and 5 (each suspension at a concentration of 800 μM) at T0+4 hours in independent and parallel experiments. In the "control" group, water was added to the wells at T0, and then at T0+4 hours. In the "A-beta" group, A-beta 1-42 was added to the wells at T0, and then water was added to the wells at T0+4 hours.

[0195] Neuronal activity was recorded as follows: - Before the addition of T0, A-beta1-42 (or water in the "control" group) At -T0+1h, T0+2h, T0+3h, T0+4h (before the addition of nanoparticles in the <<nanoparticles>> group, or "water" in the control group), T0+5h, and T0+6h.

[0196] The values ​​were obtained from 60-second bin data taken during a 30-minute period following 30 minutes of activity stabilization.

[0197] After recording at T0+6h, all wells were activated with one electrode that actively spiked them with electrical stimulation. Stimulation was performed for 30 minutes (one electrode stimulation per well in a 48-well MEA, minimum stimulation duration = 100 μs, artifact removal 2 ms after pulse, pulse 10 × biphasic + / - 500 mV). The response of the unstimulated electrode was averaged and normalized to pre-stimulation activity (Figure 10).

[0198] Microelectrode array neurochip We purchased 48-well microelectrode array neurochips from Axion Biosystems Inc. These chips have 16 passive electrodes per well. The surface was coated with polyethyleneimine (PEI, 50% in borate buffer) for 1 hour, washed, and air-dried.

[0199] Multichannel recording and multiparametric data analysis A multi-channel MAESTRO recording system from Axion Biosystems (USA) was used for recording. For extracellular recording, 48 wells of MEA were placed in the MAESTRO recording station and maintained at 37°C. Recording was performed in DMEM / 10% thermoinactivated horse serum. pH was maintained at 7.4 by a continuous flow of filtered, humidified air containing 10% CO2. Action potentials or "spikes" were recorded as spike trains and clustered into so-called "bursts." 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 start and end of short spike events.

[0200] Multiparametric high-content analysis of network activity patterns extracted 204 activity-descriptive spike column parameters. These parameters allow for accurate descriptions of activity changes in the following four categories: overall activity, burst structure, oscillation behavior, and synchronous behavior.

[0201] The functional effects of amyloid-beta 1-42 in neuronal networks exposed to electrical stimulation, and the salvage efficacy of the nanoparticles of the present invention in the neuronal networks exposed to electrical stimulation, were evaluated through the above parameters (some of which are summarized in Table 5 below).

[0202] [Table 5]

[0203] Network activity under stimulation was normalized to the relevant spontaneous native activity (recorded at T0+6 hours) set to 100% for each experiment. Values ​​related to spontaneous native activity were obtained from 60-second bin data taken from a 30-minute period after 30 minutes of activity stabilization. Results (parameter values) were expressed as the mean ± SEM of independent networks. Each "nanoparticle" group included at least 9 active wells in the analysis, the "control" group at least 18 active wells, and the "A-beta" group at least 18 active wells ("active" means a well with a sufficient number of electrodes to measure electrical activity). The absolute parameter distribution was tested for normality, and statistical significance between groups was assessed by one-way ANOVA.

[0204] Figure 11 shows some representative parameters from the following categories: overall activity, burst structure, vibration behavior, and synchronism.

[0205] These parameters characterize the A-beta 1-42 induced functional effects under electrical stimulation and the rescue efficacy enabled by the nanoparticles of the present invention under electrical stimulation (i.e., the ability to prevent / rescue functional effects to a level similar to that of the "control" group).

[0206] To evaluate the effects of the compounds, the multiparametric results of selecting 204 parameters were expressed as a single parameter called the "effect score." This is a linear combination of the selected features, transforming the dataset into a vector with a "control" group exposed to the electric field with a mean of "0" and an "A-beta" group exposed to the electric field with a mean of "1." The calculation of the Z-factor of the effect score was measured to find the best distinction between the "control" and "A-beta" groups, and was performed through the selection of 15 features out of the 204 optimized parameters (Kummel A, et al. J Biomol Screen., 2010, 15(1),95-10: Integration of multiple readouts into the z' factor for assay quality assessment). The effect score analysis is shown in Figure 12.

[0207] Table 6 shows the rescue efficacy of the nanoparticles of the present invention after exposure to electrical stimulation.

[0208] [Table 6]

[0209] Clinical studies are underway to evaluate the potential of DBS for the treatment of Alzheimer's disease. Typical stimulation parameters usable in the context of the present invention, in combination with the nanoparticles described herein, are a frequency of 130 Hz, a pulse width of 60 or 90 μs, and an amplitude voltage of 3–5 V. In the experiments described herein, stimulation was applied to a neuronal network coculture for 30 minutes. Stimulation = 10 biphase pulses, minimum pulse duration = 100 μs, pulse amplitude = + / - 500 mV, pulse frequency = 20 Hz, and pulse train duration = 0.2 Hz.

[0210] Figures 11 and 12 and Table 6 show that treatment of neuronal networks with the nanoparticles of the present invention rescues the A-beta 1-42 induced functional effects of neuronal networks under an electric field when exposed to an electric field. Interestingly, the rescue efficacy was observed for parameters in categories related to oscillatory behavior and synchronousness, and could reach levels observed in the "control" group.

[0211] These vibrational behaviors and synchronization parameters are typically monitored as measures of altered network development. These parameters can be advantageously rescued in the presence of the nanoparticles of the present invention, which are exposed to electrical stimulation.

[0212] These results highlight the advantageous performance of the nanoparticles described herein in rescuing A-beta 1-42 induced functional effects in neuronal networks under electric field conditions.

Claims

1. Nanoparticles or nanoparticle aggregates, when exposed to an electric field, are nanoparticles or nanoparticle aggregates for use in the prevention or treatment of neurological disorders or at least one symptom thereof in a subject, Materials consisting of nanoparticles or nanoparticle aggregates include conductive materials, semiconductor materials, and materials with a dielectric constant ε of 200 or more. ｉｊｋ An insulating material having a dielectric constant ε of 100 or less ｉｊｋ Nanoparticles or nanoparticle aggregates selected from insulating materials having [a certain property].

2. Nanoparticles or nanoparticle aggregates for use according to claim 1, wherein an electric field is applied by deep brain stimulation, transcranial electrical stimulation, or transcranial magnetic stimulation.

3. The nanoparticles or nanoparticle aggregates for use according to claim 1 or 2, 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 adjacent sp2 hybridized carbon centers in their structure.

4. The nanoparticles or nanoparticle aggregates for use according to claim 3, wherein the material of the nanoparticles or nanoparticle aggregates is selected from metal nanoparticles or mixtures thereof in which the metal element is Ir, Pd, Pt, Au, and organic nanoparticles consisting of polyaniline, polypyrrole, polyacetylene, polythiophene, polycarbazole and / or polypyrene.

5. The nanoparticles or nanoparticle aggregates for use according to claim 1 or 2, wherein the material of the nanoparticles or nanoparticle aggregates is a semiconductor material having a band gap Eg of less than 3.0 eV.

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

7. The nanoparticles or nanoparticle aggregates for use according to claim 6, wherein the material of the nanoparticles or nanoparticle aggregates consists of elements from Group IVA of the Mendeleev periodic table and is doped with charge carriers selected from Al, B, Ga, In, and P.

8. The material is an insulating material having a band gap Eg of 3.0 eV or more, and the relative permittivity ε ｉｊｋ However, 20°C to 30°C, 10 ２ Nanoparticles or nanoparticle aggregates for use according to claim 1 or 2, which are measured between Hz and infrared frequencies.

9. The material is an insulator material having a band gap Eg of 3.0 eV or more, and a relative permittivity 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 The dielectric material which is a mixed metal oxide selected from is a nanoparticle or nanoparticle aggregate for use according to claim 8.

10. The material is an insulating material having a band gap Eg of 3.0 eV or more, and the relative permittivity ε ｉｊｋ The nanoparticles or nanoparticle aggregates for use according to claim 8, wherein the nanoparticles or nanoparticle aggregates are less than or equal to 100, and the material of the nanoparticles or nanoparticle aggregates is a dielectric material selected from metal oxides, mixed metal oxides, and carbon materials, wherein the metal element is a metal element or lanthanide from period 3, 5, or 6 of the Mendeleev periodic table.

11. Nanoparticles for use according to any one of claims 1 to 10, wherein the neurological disorder is selected from Parkinson's disease, Alzheimer's disease, epilepsy, obsessive-compulsive disorder, autism spectrum disorder, depressive disorder, dystonia, Tourette 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.

12. A composition for use in the prevention or treatment of neurological disorders or at least one symptom thereof in an object exposed to an electric field, The composition comprises nanoparticles and / or nanoparticle aggregates and a pharmaceutically acceptable support, wherein the material of the nanoparticles or nanoparticle aggregates is a conductive material, a semiconductor material, or a dielectric constant ε of 200 or more. ｉｊｋ An insulating material having a dielectric constant ε of 100 or less ｉｊｋ A composition selected from insulating materials having [a certain property].

13. The composition comprises at least two distinct nanoparticles and / or nanoparticle aggregates, each nanoparticle or nanoparticle aggregate being a conductive material, a semiconductor material, and having a dielectric constant ε of 200 or more. ｉｊｋ An insulating material having a dielectric constant ε of 100 or less ｉｊｋ A composition for use according to claim 12, comprising a separate material selected from insulating materials having the property.

14. It comprises at least two distinct nanoparticles and / or nanoparticle aggregates, each nanoparticle or nanoparticle aggregate being a conductive material, a semiconductor material, and having a dielectric constant ε of 200 or more. ｉｊｋ An insulating material having a dielectric constant ε of 100 or less ｉｊｋ A kit consisting of separate materials selected from insulating materials having [specific properties].

15. The kit according to claim 14, for use in the prevention or treatment of a neurological disorder or at least one symptom thereof in a subject.

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