Monitoring tissue permeability during ultrasound procedures

Abstract

The present disclosure relates, in part, to targeted drug delivery using ultrasound procedures, and more particularly to systems and methods for measuring drug delivery levels and certain aspects of metabolism, such as drug metabolism, in brain tissue.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 322,617, filed on March 22, 2022, the entire contents of which are incorporated herein. Technical Field

[0003] The present invention relates generally to targeted drug delivery and, more particularly, to systems and methods for enhancing targeted drug delivery using ultrasound procedures. Background Art

[0004] The evaluation of drug delivery efficiency is very important for making key clinical decisions. Biomarkers that provide insights into indicators of normal or pathological biological processes are routinely analyzed to evaluate responses to drugs and other medical interventions. Liquid biopsy is a non-invasive technique for sampling blood from subjects to detect and measure biomarkers. The analysis results after liquid biopsy (e.g., evaluation of the amount and / or activity of proteins / enzymes, metabolites, nucleic acids, karyotypes, mutations, genetic markers, etc.) can infer disease progression, efficacy of drugs or other interventions, stability and metabolism of drugs, etc. Therefore, liquid biopsy is very helpful for diagnosis, selection of therapy, determination of dosage and dosage regimen, efficacy, etc. The use of liquid biopsy has made progress in a variety of diseases including genetic disorders, cancer, infectious diseases, metabolic diseases, environmental diseases, allergies, autoimmune disorders, etc. However, its use in certain tissue diseases is limited by the lack of permeability of biomarkers from those tissues to the blood. For example, the blood-brain barrier ("BBB") prevents biomarkers from being transported to the blood from the central nervous system (CNS). Therefore, without the use of more sophisticated tests, the diagnosis and treatment of diseases such as stroke, multiple sclerosis (MS), neurodegenerative disorders, Alzheimer's disease (AD), Parkinson's disease, epilepsy and traumatic brain injury are challenged. Other such barriers that impede the transfer of biomarkers from tissue to blood include the blood-testis barrier, the blood-retina barrier, the blood-placental barrier and the blood-thymus barrier.

[0005] The goal of developing a drug delivery system is usually to reduce the overall therapeutic dose of the drug, increase its residence time, extend its release over time, and enhance targeting to diseased tissues. The above barriers hinder the targeted delivery of drugs. A conventional method for enhancing targeted drug delivery involves the use of ultrasound. For example, exposing the target tissue to ultrasound may increase its permeability, thereby allowing a larger proportion of the drug dose to exert therapeutic effects on the target area. In another method, microbubbles are injected into the bloodstream to enhance the permeability of blood vessels to various drugs through ultrasound and biomechanical interactions with microbubbles. In addition, drugs can be encapsulated in "nanoparticles" and injected into the target area through microbubbles; the application of ultrasound causes the microbubbles to undergo structural mechanical deformation and / or temperature increase, thereby triggering the release of encapsulated drugs. Similarly, ultrasonic energy can be applied to chemically activate drugs applied to the target area.

[0006] Specifically, many studies have explored the safety and efficacy of using ultrasound to open the blood-brain barrier (BBB) ​​to allow the passage of neurotherapeutics. In general, the BBB blocks molecules with a molecular weight above about 400 Daltons from entering the brain parenchyma, rendering the vast majority of neurotherapeutics and biomarkers ineffective. Ultrasound has been shown to be a safe and effective BBB opening mediated by intravascular microbubble oscillations. Its advantages include its non-invasive nature, its exceptional millimeter-level precision, and the reversibility of its effects on the BBB.

[0007] The combination of ultrasound and liquid biopsy makes it possible to study some central nervous system biomarkers. However, even if the BBB has changed, its permeability still has significant heterogeneity, which hinders the reliable detection and / or measurement of biomarkers. For example, the amount of biomarkers in two measurements of the same subject is not equivalent to each other. This hinders the efforts to monitor disease progression, reliably deliver accurate amounts of drugs, and monitor the efficacy of clinical interventions. Therefore, the industry still needs to be able to predict the extent to which ultrasound (or similar therapies) open the barrier, the extent of drug delivery, monitor changes in biomarkers, and compare one application of ultrasound (or similar therapies) to open the barrier with another application. Without accurate predictions, it is difficult to make key decisions about therapy. Summary of the invention

[0008] The present invention provides systems and methods for evaluating tissue permeability enhancement in a target area using an ultrasound procedure. For example, a target tissue is generally permeable to molecules less than 400 Daltons in size, but a therapeutic agent may be 1,000 Daltons or larger in size and therefore blocked from entering the target area. When ultrasound is applied, tissue in the target area or tissue that regulates access to the target area may be damaged, and therefore its permeability may increase. The level of permeability and / or the size of the tissue area with increased permeability may depend on the intensity and / or duration of ultrasound application. Therefore, by adjusting ultrasound parameters, the tissue permeability of the target area can be increased to a desired degree to allow the therapeutic agent to penetrate and / or diffuse therein. The present disclosure also provides systems and methods for standardizing permeability between instances of ultrasound treatment to allow an accurate amount of drug to be delivered to the target tissue each time.

[0009] Several techniques for monitoring target tissue permeability during or after ultrasound exposure are presented herein. Generally, they involve the use of molecules or molecular complexes of a similar size range to the therapeutic agent of interest, and the passage of the molecules or molecular complexes through the disrupted target tissue can be observed and quantified.

[0010] In the first approach, the relaxation rate It is a physical parameter measured regularly in MRI systems as a safety metric to measure and predict the degree of BBB opening during treatment. For example, Can be used to predict liposome concentration. Liposomes can be conjugated with MRI-visible substances such as Gd ions (see, e.g., "Nonlinear Effects in Perfusion Quantification Using Bolus-Tracking MRI," Mag. Res. In Medicine, 61: 486-492 (2009), which is incorporated herein by reference), but such contrast agents are not necessary for this method. Alternatively, the transverse relaxation time can be used To replace or supplement Again, contrast medium is not necessary.

[0011] Another MRI parameter that can be used to assess tissue permeability is the longitudinal relation time T1, which is a measure of the time required for rotating protons to realign with the external magnetic field during the MRI procedure. They can be used in combination to estimate tissue permeability levels, or can be used independently as a measure of permeability (although T1 measurements may be obtained only once in a given treatment period).

[0012] In another method, the permeability of BBB to different molecules is predicted based on the entry of nanodroplets or liposomes of similar size filled with one or more acoustic agents. For example, sonodynamic therapy is a non-thermal ultrasound technology that utilizes the sonochemical action of cavitation bubbles. Rose Bengal (RB) has sonochemical activity and can reduce the cavitation threshold, so sonodynamic therapy can be supported. Other sonodynamic agents include nanomaterials (particularly TiO2) based on titanium (Ti), titanium hydride, titanium carbide, xanthene dyes (such as fluorescein, tetrachlorofluorescein, eosin Y, erythrosine B and fluorescent red B), protoporphyrin IX, hematoporphyrin monomethyl ether (HMME), dihydrochlorin e6 (Ce6) and nanoparticles based on silicon. These agents can be encapsulated in liposomes by reverse evaporation and extrusion. An alternative method is to use acoustic response phase shift nanodroplets. The size of these agents and other detection agents (but not limited to, for example, magnetic resonance imaging (MRI) visible contrast agents containing the ion gadolinium (Gd)) can be adjusted by complexing (e.g., by chemical conjugation) with entities such as polymers (but not limited to, for example, polyethylene glycol and dextran) or encapsulation in liposomes. Therefore, these agents and other detection agents can be used to standardize therapeutic agents to a desired molecular weight range.

[0013] In another approach, the permeability of the BBB to different molecules is predicted based on the biomarkers of similar size leaving the target tissue and entering the bloodstream. Non-limiting examples of target tissue-specific biomarkers include central nervous tissue-specific markers, such as neuron-specific enolase (NSE) and S100 calcium-binding protein B (S100B), which are not normally found in the blood.

[0014] Thus, in a first aspect, a system for destroying target tissue for treatment is provided. The system includes an ultrasonic transducer for sonicating a target volume to cause destruction of the target tissue therein, thereby increasing its permeability. The system also includes a controller configured to (i) monitor the permeability of the target tissue caused by the destruction, and (ii) stop the ultrasonic transducer from sonicating the target volume when the permeability reaches a threshold value.

[0015] In some embodiments, permeability is calculated based on the measurement of molecules present (and in some cases mainly present) in the target tissue under normal conditions but not substantially present in tissues outside the target tissue. "Substantially absent" means that the background level of the molecule is small enough that its presence increased by barrier permeability can be easily detected. In some embodiments, the target tissue is the brain. In some embodiments, the molecule is mainly present in the target tissue under normal conditions, but is not substantially present in tissues outside the target tissue. Illustrative, non-limiting examples of molecules present in the target tissue but not in tissues outside the target tissue include NSE and S100B. In some embodiments, permeability is calculated based on the measurement of detection markers in the target tissue, and the detection markers are applied to the subjects in which the permeability is calculated. In some embodiments, the detection markers and / or are compounded with polymers, liposomes, quantum dots, dextran, etc. In some embodiments, the detection marker is an MRI visible contrast agent, which optionally includes ion Gd.

[0016] In some embodiments, the permeability corresponds to the upper limit of the size distribution of molecules or molecular complexes that pass through the destroyed target tissue. The permeability may alternatively or additionally correspond to the concentration of molecules or molecular complexes that have passed through the destroyed target tissue. In some embodiments, the permeability is calculated based on the measurement of molecules within a certain size range and present in the target tissue but not in tissues outside the target tissue.

[0017] In some embodiments, the permeability is compared between at least two sonication applications administered to the same subject on different days. In some embodiments, the permeability is compared between at least two sonication applications administered to different subjects.

[0018] In some embodiments, permeability is standardized using a detection agent or molecule that is mainly present in the target tissue under normal conditions but is substantially absent in tissues outside the target tissue. In some embodiments, the target tissue is the brain, and molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue. Illustrative, non-limiting examples of molecules that are mainly present in the target tissue but are substantially absent in tissues outside the target tissue include, but are not limited to, brain proteins NSE, S100B, astrocyte fiber actin (GFAP) and neuronal ubiquitin carboxyl terminal hydrolase isozyme L1 (UCH-L1), myelin basic protein (MBP), tau, and neurofilament light chain (NfL). In some embodiments, the detected NSE species has a molecular weight of about 78kDa. In some embodiments, the detected S100B species has a molecular weight of about 22kDa. In some embodiments, measurement of NSE and / or S100B species is used to normalize the delivery of therapeutic agents having molecular weights ranging from about 60 kDa to about 100 kDa and / or about 15 kDa to about 30 kDa, respectively.

[0019] In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent. In some embodiments, the detection marker is complexed with a liposome having a defined size range corresponding to the size range of the therapeutic agent.

[0020] The size range of such molecules or molecular complexes may correspond to monoclonal antibodies, viral vectors, liposomes, nucleic acids (e.g., miRNA, mRNA, etc.), or proteins. Some embodiments use markers of relevant sizes. Some embodiments select markers (e.g., monoclonal antibodies, viral vectors, liposomes, proteins, and / or nucleic acids) that are comparable in size to the desired drug. The size of the molecule or complex may be similar to the drug of interest, but have minimal therapeutic effect when administered.

[0021] In some embodiments, the system further comprises an MRI device. The controller may be responsive to the MRI device and may be based on T1 or The permeability can be assessed based on at least one measurement of the contrast agent. The relationship between the measurement and the permeability can be established by calibration, and in some embodiments, the calibration can include a machine learning component. Machine learning can alternatively or additionally be used to improve the calibration curve over time. The controller can be configured to assess the permeability in the absence of a contrast agent.

[0022] In some embodiments, the target species corresponds to a molecule or molecular complex coupled to a visualization agent. The visualization agent can be a contrast agent or a fluorophore, such as ion Gd. The molecule or molecular complex can be a liposome, quantum dot, or dextran.

[0023] In some embodiments, the transducer is also configured to detect an acoustic signal. This is not an acoustic response during therapeutic ultrasound treatment, but rather a diagnosis after ultrasound treatment. The controller can respond to the acoustic signal detected by the transducer and can assess the permeability based on acoustic measurements of the target species. The target species can correspond to a molecule or molecular complex coupled to the acoustic agent. The controller can be further configured to assess the permeability based at least in part on a combination of acoustic measurements and MRI images. The controller can update the target acoustic dose level based at least in part on a combination of acoustic measurements and MRI images.

[0024] In various embodiments, the controller is configured to control ultrasound therapy based at least in part on the updated target acoustic dose level. The therapy may include selecting an initial desired acoustic dose, performing ultrasound therapy using the desired acoustic dose and measuring the acoustic dose, calculating MRI (e.g., ) map, updates the required acoustic dose based on the MRI, and controls the next ultrasound treatment based on the revised acoustic dose. Some embodiments update the acoustic dose after each ultrasound treatment during treatment, and once the controller has established what works for a particular patient, it can use that acoustic dose in future treatment sessions. Different values ​​( Once updated, the target acoustic dose level can be reused during treatment or even during the next treatment session. This may cover situations where the patient is treated in the MRI and then goes home, and may alleviate the need for MRI guidance during the next treatment session. The parameters of subsequent treatment sessions may be changed based on the parameters of the previous treatment and the observed clinical outcomes (e.g., increasing the dose in case of an inadequate response).

[0025] The permeability can correspond to the concentration of agent-coupled molecules or molecular complexes having a target size range and having passed through the target tissue to be destroyed. The visualization agent can be a contrast agent, and the concentration can be determined at least in part by a wash-in time or a wash-out time. The wash-out time can correspond to a rise or fall in an acoustic signal characteristic relative to a threshold. The acoustic agent can be a contrast agent, and the signal characteristic can be the reflected amplitude of an acoustic signal emitted by a transducer.

[0026] In another aspect, a method for destroying a target tissue for treatment is provided. In various embodiments, the method includes ultrasonicating a target volume to cause damage to the target tissue therein, thereby increasing its permeability; monitoring the permeability of the target tissue caused by the damage; and stopping the ultrasonication of the target volume when the determined permeability reaches a threshold value.

[0027] In some embodiments, permeability is calculated based on: (i) measurement of the level of a molecule that is predominantly present in the target tissue under normal conditions but substantially absent in tissues outside the target tissue in a biological sample from the subject in which the permeability is being monitored, and / or (ii) measurement of a detection marker within the target tissue, wherein the detection marker has been administered to the subject in which the permeability is being monitored.

[0028] In some embodiments, the target tissue is brain. In some embodiments, the biological sample is blood. In some embodiments, the molecule is mainly present in the target tissue under normal conditions, but is substantially absent in tissues outside the target tissue. Illustrative, non-limiting examples of molecules that are mainly present in the target tissue but are substantially absent in tissues outside the target tissue include, but are not limited to, brain proteins NSE, S100B, GFAP and UCH-L1, MBP, tau and NfL. In some embodiments, the detected NSE species has a molecular weight of about 78kDa. In some embodiments, the detected S100B species has a molecular weight of about 22kDa. In some embodiments, the measurement of NSE and / or S100B species is used to standardize the transport of therapeutic agents with a molecular weight range of about 60kDa to about 100kDa and / or about 15kDa to about 30kDa, respectively.

[0029] In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent or is complexed to a liposome having a defined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is an MRI visible contrast agent, which optionally includes ion Gd. In some embodiments, the sonication comprises at least two applications to the same patient on different days or different applications to different patients.

[0030] In another aspect, a method for comparing the permeability of a target tissue in a subject between at least two focused ultrasound treatment applications on different days is provided, the method comprising: (i) providing a biological sample from a subject who received a first ultrasound treatment application to a target volume to cause damage to the target tissue therein, thereby increasing its permeability, (ii) measuring the amount of a molecule in the biological sample that under normal conditions is primarily present in the target tissue but is substantially absent in tissues outside the target tissue, (iii) monitoring the permeability of the target tissue caused by the damage; (iv) providing a second biological sample from a subject who received a second ultrasound treatment application to the target volume on a different day to cause a second damage to the target tissue therein, thereby increasing its second permeability, (v) measuring the amount of a molecule in the second biological sample that under normal conditions is primarily present in the target tissue but is substantially absent in tissues outside the target tissue, (vi) monitoring the permeability of the target tissue caused by the second damage; and (vii) comparing the permeability of the target tissue in the subject between the at least two ultrasound treatment applications on different days.

[0031] In some embodiments, the target tissue is brain and / or the biological sample is blood. In some embodiments, the molecule that is mainly present in the target tissue but is substantially absent in the tissue outside the target tissue under normal conditions is brain protein. In some embodiments, brain protein is usually not found outside the skull tissue (for example, in the bloodstream). In some embodiments, when the blood-brain barrier is fully opened to allow the leakage of molecules with the size or molecular weight of brain protein, brain protein will leak outside the skull tissue (for example, in the bloodstream). In some embodiments, when the blood-brain barrier is fully opened to allow the leakage of molecules with the size or molecular weight of brain protein, brain protein becomes detectable in biological samples (such as blood samples). Illustrative, non-limiting examples of molecules that are mainly present in the target tissue but are substantially absent in the tissue outside the target tissue include but are not limited to brain protein NSE, S100B, GFAP and UCH-L1, MBP, tau and NfL. In some embodiments, the detected NSE species has a molecular weight of about 78kDa. In some embodiments, the detected S100B species has a molecular weight of about 22kDa. In some embodiments, the GFAP species detected are monomers (50 kDa), dimers (100 kDa) and / or tetramers (200 kDa). Tang et al., J Biol Chem. Oligomers of Mutant Glial Fibrillary Acidic Protein (GFAP) Inhibit the Proteasome System in Alexander Disease Astrocytes, and the Small Heat Shock Protein αB-Crystallin Reverses the Inhibition, 2010; 285 (14): 10527-10537. Similarly, in some embodiments, UCH-L1, MBP, tau, and NfL species have molecular weights of about 25 kDa, about 18,500 Daltons, about 62 kDa, and about 68 kDa, respectively. Thus, in some embodiments, measurement of NSE species is used to standardize the delivery of therapeutic agents with molecular weights ranging from about 60 kDa to about 100 kDa. In some embodiments, the measurement of S100B species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 15 kDa to about 30 kDa. In some embodiments, the measurement of GFAP species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 40 kDa to about 250 kDa, depending on the species detected. In some embodiments, the measurement of UCH-L1 species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 20 kDa to about 30 kDa.In some embodiments, the measurement of MBP species is used to standardize the transport of therapeutic agents with a molecular weight range of about 15 kDa to about 20 kDa. In some embodiments, the measurement of tau species is used to standardize the transport of therapeutic agents with a molecular weight range of about 50 kDa to about 70 kDa. In some embodiments, the measurement of NfL species is used to standardize the transport of therapeutic agents with a molecular weight range of about 60 kDa to about 80 kDa. In various embodiments, the measurement of one or more molecular species that are primarily present in the target tissue but are substantially absent in tissues outside the target tissue is used to standardize the transport of therapeutic agents with a molecular weight range of about 15 kDa to about 250 kDa.

[0032] In another aspect, a method for comparing the permeability of a target tissue in a subject between at least two focused ultrasound treatment applications on different days is provided, the method comprising: (i) providing a subject who has received a first ultrasound treatment application to a target volume to cause damage to the target tissue therein, thereby increasing its permeability, wherein the subject has optionally received a detection marker on the same day, (ii) measuring the level of the detection marker in the target tissue, thereby monitoring the permeability of the target tissue caused by the damage; (iii) providing a second biological sample from the subject who has received a second ultrasound treatment application to the target volume on a different day to cause a second damage to the target tissue therein, thereby increasing its second permeability, optionally wherein the subject has received a detection marker on the same day, (iv) measuring the level of the detection marker in the target tissue, thereby monitoring the permeability of the target tissue caused by the second damage, thereby monitoring the permeability of the target tissue caused by the second damage; and (v) comparing the permeability of the target tissue in the subject between the at least two ultrasound treatment applications on different days.

[0033] In some embodiments, the target tissue is the brain. In some embodiments, the target tissue is the brain, and the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol), which is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent. In some embodiments, the target tissue is the brain, and the detection marker is complexed with a liposome, which has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is an MRI visible contrast agent. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the detection marker is a positron emission tomography (PET) reporter. In some embodiments, the PET reporter is selected from 18F-Fluorbetapyr (18F-florbetapir), 18F-Fluorbetaben (18F-Florbetaben), 18F-Fluortaucipir (18F-Flortaucipir), 18F-Flutemetamol (18F-Flutemetamol) and 18F-Fluordopa (18F-Flourodopa, F-Dopa). In some embodiments, the detection marker is a single photon emission computed tomography (SPECT) reporter. In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane and 99m Tc-TRODAT-1.

[0034] In another aspect, the present invention is directed to a method of treating a neurological disease or disorder in a subject in need thereof, wherein the neurological disease or disorder is characterized by a site in the brain having abnormal production, aggregation and / or deposition of a protein or another biomolecule, wherein a therapeutic agent and / or a microbubble composition will be, is being, or has been administered to the subject, the method comprising the steps of: sonicating a target volume to cause damage to target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and an adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the damage; and when the permeability reaches a threshold value, ceasing sonication of the target volume, thereby increasing the level of delivery of the therapeutic agent to the site compared to a control.

[0035] In some embodiments, the control is the delivery level of the therapeutic agent in a subject that has not yet received the acoustic pulse sequence and / or microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in a subject before the administration of the acoustic pulse sequence and / or microbubble composition.

[0036] In some embodiments, the nervous system disease or disorder is selected from Alzheimer's Disease (AD), Parkinson's Disease (PD), Huntington's Disease (HD), amyotrophic lateral sclerosis (ALS), dementia with Lewy bodies, spinocerebellar ataxia and amyotrophic lateral sclerosis, frontotemporal lobar disease, multiple system atrophy, four-repeat tau disease and prion disease. In some embodiments, the site is selected from senile plaques, neurofibrillary tangles, neuronal inclusions, Lewy bodies, glial inclusions, cytoplasmic inclusions and polyglutamine aggregates. In some embodiments, the protein showing abnormal production, aggregation and / or deposition is selected from amyloid-β (Aβ), Tau protein, TDP-43, α-synuclein, FUS / TLS, SOD1 and Huntingtin.

[0037] In some embodiments, the therapeutic agent includes a small molecule or a biological drug. In some embodiments, the therapeutic agent is or includes a biological drug. In some embodiments, the therapeutic agent is selected from a gene therapy agent, an enzyme for enzyme replacement therapy (e.g., ceramidase, acid α-glucosidase (EC 3.2.1.20), agalsidase β, α-l-iduronidase, acid sphingomyelinase, imiglucerase, glucosylceramide (GlcCer) synthase, and arylsulfatase A (ARSA); the replaced enzyme can be administered as a purified protein, a nucleic acid encoding an active enzyme, and / or as a nucleic acid that corrects a defect (e.g., regulates correct splicing), a vaccine, an antisense oligonucleotide (ASO), a protein therapeutic, a modified mRNA agent, and an RNAi agent. In some embodiments, the therapeutic agent is or includes an antibody, an antibody-like molecule, or an antigen-binding fragment thereof . In some embodiments, the therapeutic agent specifically binds to a protein or another biomolecule that exhibits abnormal production, aggregation, and / or deposition. In some embodiments, the therapeutic agent is selected from a nonspecific clearance antibody (e.g., intravenous immunoglobulin, also known as IVIg), an anti-amyloid-β antibody (e.g., aducanumab, gantenerumab, lecanemab, and donanemab), an anti-tau antibody (e.g., se morinemab, gosuranemab, tilavonemab, and zagotenemab), an anti-TREM2 antibody (e.g., AL002), an anti-α-synuclein antibody (e.g., Cinpanemab, Prasinezuma b, Lu AF82422, ABBV-0805, and MEDI1331), and / or a combination thereof.

[0038] In some embodiments, the therapeutic agent is or includes a small molecule drug. In some embodiments, the therapeutic agent provides synaptic plasticity, neuroprotection, reduced inflammation, neurotransmitter receptor regulation, reduced oxidative stress in one or more. In some embodiments, the therapeutic agent is selected from donepezil, galantamine, rivastigmine, memantine, suvorexant, carbidopa-levodopa, selegiline, rasagiline, safinamide, entacapone, benztropine, tolcapone, opicapone, nuplazid, istradefylline and amantadine and combinations thereof.

[0039] In some embodiments, the therapeutic agent is formulated in liposomes. In some embodiments, the therapeutic agent is delivered via a viral vector.

[0040] In another aspect, the present invention relates to a method for treating a neurological disease or condition in a subject in need thereof, wherein the neurological disease or condition is characterized by a site in the brain having abnormal production, aggregation and / or deposition of a protein or another biomolecule, the method providing a predetermined range of local doses of a therapeutic agent, the method comprising the following steps: (i) subjecting a target volume to ultrasound treatment to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and the adjacent blood-brain barrier (BBB), wherein the subject has received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker, optionally on the same day; (ii) measuring the level of the detection marker in the target tissue, thereby monitoring the local dose of the therapeutic agent caused by the damage at the site; (iii) stopping ultrasound treatment of the target volume when the local dose of the therapeutic agent reaches a predetermined range.

[0041] In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome having a defined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a magnetic resonance imaging (MRI) visible contrast agent, a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0042] In some embodiments, the neurological disease or disorder is selected from Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), dementia with Lewy bodies, spinocerebellar ataxia and amyotrophic lateral sclerosis, frontotemporal lobar disease, multiple system atrophy, four-repeat tau disease and prion disease. In some embodiments, the site is selected from senile plaques, neurofibrillary tangles, neuronal inclusions, Lewy bodies, glial inclusions, cytoplasmic inclusions and polyglutamine aggregates. In some embodiments, the protein showing abnormal production, aggregation and / or deposition is selected from amyloid-β (Aβ), Tau protein, TDP-43, α-synuclein, FUS / TLS, SOD1 and Huntington protein.

[0043] In some embodiments, the therapeutic agent comprises a small molecule or a biological drug. In some embodiments, the therapeutic agent is or comprises a biological drug. In some embodiments, the therapeutic agent is selected from a gene therapy agent, an enzyme for enzyme replacement therapy (e.g., ceramidase, acid α-glucosidase (EC 3.2.1.20), agalsidase β, α-l-iduronidase, acid sphingomyelinase, imiglucerase, glucosylceramide (GlcCer) synthase, and arylsulfatase A (ARSA); the replaced enzyme can be administered as a purified protein, a nucleic acid encoding an active enzyme, and / or as a nucleic acid that corrects a defect (e.g., regulates correct splicing), a vaccine, an antisense oligonucleotide (ASO), a protein therapeutic, a modified mRNA agent, and an RNAi agent. In some embodiments, the therapeutic agent is or comprises an antibody, an antibody-like molecule, or an antigen-binding fragment thereof. In some embodiments In some embodiments, the therapeutic agent specifically binds to a protein or another biomolecule that exhibits abnormal production, aggregation, and / or deposition. In some embodiments, the therapeutic agent is selected from a nonspecific clearance antibody (e.g., intravenous immunoglobulin, also known as IVIg), an anti-amyloid-β antibody (e.g., aducanumab, gantuzumab, lencanezumab, and donenetumab), an anti-tau antibody (e.g., ceranatumab, gonelumab, tipranezumab, and zagetelumab), an anti-TREM2 antibody (e.g., AL002), an anti-α-synuclein antibody (e.g., simpanezumab, putinizumab, Lu AF82422, ABBV-0805, and MEDI1331), and / or a combination thereof.

[0044] In some embodiments, the therapeutic agent is or includes a small molecule drug. In some embodiments, the therapeutic agent provides one or more of synaptic plasticity, neuroprotection, reduced inflammation, neurotransmitter receptor modulation, and reduced oxidative stress. In some embodiments, the therapeutic agent is selected from donepezil, galantamine, rivastigmine, memantine, suvorexan, carbidopa-levodopa, selegiline, rasagiline, safinamide, entacapone, benztropine, tolcapone, opicapone, nuptazide, istradefylline, and amantadine, and combinations thereof.

[0045] In some embodiments, the therapeutic agent is formulated in liposomes. In some embodiments, the therapeutic agent is delivered via a viral vector.

[0046] As used herein, the term "substantially" means ± 10% of tissue volume and in some embodiments means ± 5% of tissue volume. "Clinically significant" means that the tissue has an unwanted effect (sometimes lacking the desired effect) that is considered significant by a clinician, such as triggering the occurrence of tissue damage. Mentioning "one embodiment", "embodiment", "one implementation" or "implementation" throughout this specification means that the specific features, structures or characteristics described in conjunction with the embodiment are included in at least one embodiment of the technology of the present invention. Therefore, the phrases "in one embodiment", "in an embodiment", "one implementation" or "implementation" that appear throughout this specification do not necessarily refer to the same embodiment. In addition, specific features, structures, conventions, steps or characteristics can be combined in one or more embodiments of the technology in any suitable manner. The titles provided herein are for convenience only and are not intended to limit or explain the scope or meaning of the claimed technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] In the accompanying drawings, the same reference numerals generally refer to the same parts in different views. In addition, the drawings are not necessarily to scale, but emphasis is generally placed on illustrating the principles of the invention. In the following description, various embodiments of the invention will be described with reference to the following drawings, in which:

[0048] Figure 1A Exemplary ultrasound systems according to various embodiments of the present invention are schematically depicted.

[0049] Figure 1B An exemplary MRI system according to various embodiments of the present invention is schematically depicted.

[0050] Figure 2 Depicted are one or more focal regions of ultrasound waves / pulses generated in a target volume according to various embodiments.

[0051] Figure 3 Depicted is the presence of microvesicles in a target tissue region according to various embodiments.

[0052] Figure 4 is a flow chart of a representative process for correlating MR parameters with trans-BBB liposome penetration according to various embodiments.

[0053] Figure 5A and Figure 5B Shown according to various embodiments The relationship between the concentration of Gd-liposomes.

[0054] Fig. 6A Shown is a graphical plot of liposome particle distribution calculated from dynamic light scattering (DLS) data according to some embodiments.

[0055] Figure 6B The relationship between R1 and Gd-liposome complex concentration observed according to some embodiments is graphically depicted.

[0056] Figure 7 A- Figure 7 C shows according to some embodiments Graph and its relationship to the sub-point positions.

[0057] Figure 8 A depicts a graph of R1 indicating the presence of Gd in 100 nm liposomes and a graphical plot of R1 against Gd concentration in rat blood; and Figure 8 B shows similar results for 50 nm liposomes. DETAILED DESCRIPTION

[0058] We first describe systems and methods for disrupting the BBB to allow delivery of therapeutic agents, and then present techniques for evaluating and monitoring induced tissue permeability.The present disclosure is based in part on the discovery of techniques for evaluating and monitoring induced tissue permeability across a tissue, such as the BBB.

[0059] The technology disclosed herein enables prediction of the degree of ultrasound (or similar therapy) opening the barrier and the degree of drug delivery. The technology disclosed herein also allows monitoring of changes in biomarkers, and compares one application of opening the barrier by ultrasound (or similar therapy) with another application. Through such predictions, the technology disclosed herein helps make key decisions, especially about selecting or changing therapy, formulating dosage and dosing regimen, monitoring the efficiency of local drug delivery, and monitoring therapeutic efficacy.

[0060] In some aspects, the technology relies on the measurement of the transport of detection markers (not limited to, for example, MRI visible contrast agents) administered to the patient to the target tissue. In these embodiments, the detection marker is transported to the target tissue after inducing permeability (not limited to, for example, using ultrasound). In some embodiments, the molecular weight or size of the detection marker is used to calculate the degree of transport of therapeutic agents with a molecular weight or size similar to the detection marker. In some embodiments, the molecular weight or size of the detection marker is used for standardization, with the purpose of comparing the degree of transport of therapeutic agents during different permeability induction applications.

[0061] In some aspects, the technology relies on the detection and measurement of target tissue-specific molecules outside the target tissue after inducing permeability (not limited to, for example, using ultrasound). In these embodiments, the molecular weight or size of the target tissue-specific molecule is used to calculate the extent of delivery of therapeutic agents with a molecular weight or size similar to the target tissue-specific molecule. In these embodiments, the molecular weight or size of the target tissue-specific molecule is used for standardization in order to compare the extent of delivery of therapeutic agents during different permeability induction applications.

[0062] In some embodiments, the permeability is calculated based on normalization of the measurement of the detection markers or biomarkers disclosed herein. In some embodiments, the permeability is calculated and compared between two or more different ultrasound applications in the same patient. In some embodiments, the permeability is calculated and compared between different patients after ultrasound application based on normalization.

[0063] 1. Use ultrasound to increase tissue permeability

[0064] Figure 1A An exemplary ultrasound system 100 is illustrated for generating and delivering a focused acoustic energy beam to a target area to damage tissue, thereby causing increased tissue permeability therein. In various embodiments, the system 100 includes a phased array 102 of transducer elements 104, a beamformer 106 that drives the phased array 102, a controller 108 that communicates with the beamformer 106, and a frequency generator 110 that provides an input electrical signal to the beamformer 106.

[0065] The array 102 may have a curved shape (e.g., spherical or parabolic) suitable for placing it on the surface of the patient's body, or may include one or more flat or other shaped portions. Its size may vary between a few millimeters and tens of centimeters. The transducer elements 104 of the array 102 may be piezoelectric ceramic elements and may be mounted in silicone rubber or any other material suitable for mechanical coupling between the damping elements 104. Piezoelectric composites may also be used, or generally any material capable of converting electrical energy into acoustic energy. To ensure maximum power transmission to the transducer elements 104, the elements 104 may be configured to be electrically resonant at 50Ω, thereby matching the input connector impedance.

[0066] The transducer array 102 is coupled to a beamformer 106 that drives the individual transducer elements 104 so that they collectively produce a focused ultrasound beam or field. For n transducer elements, the beamformer 106 may contain n drive circuits, each drive circuit including or consisting of an amplifier 118 and a phase delay circuit 120; each drive circuit drives one of the transducer elements 104. The beamformer 106 receives a radio frequency (RF) input signal, typically in the range of 0.1 MHz to 10 MHz, from a frequency generator 110, which may be, for example, a model DS345 generator available from Stanford Research Systems. For the n amplifiers 118 and delay circuits 120 of the beamformer 106, the input signal may be divided into n channels. In some embodiments, the frequency generator 110 is integrated with the beamformer 106. The RF generator 110 and the beamformer 106 are configured to drive the individual transducer elements 104 of the transducer array 102 at the same frequency but at different phases and / or different amplitudes.

[0067] The amplification or attenuation factors α1-αn and phase shifts a1-an applied by the beamformer 106 are used to transmit and focus the ultrasonic energy on the target area and to explain the wave distortion caused in the tissue between the transducer element 104 and the target area. The amplification factor and phase shift are calculated using a controller 108, which can provide the calculation function through software, hardware, firmware, hard wiring or any combination thereof. For example, the controller 108 can utilize a general or dedicated digital data processor programmed with software in a conventional manner without too much experimentation to determine the phase shift and amplification factor required to obtain the desired focus or any other desired spatial field pattern in the target area. In some embodiments, the calculation is based on detailed information about the characteristics of the tissues (e.g., structure, thickness, density, etc.) located between the transducer elements 104 and their effects on the propagation of acoustic energy. Such information can be obtained from an imager 122. The imager 122 can be, for example, an MRI device, a computed tomography (CT) device, a positron emission tomography (PET) device, a single photon emission computed tomography (SPECT) device, or an ultrasound examination device. The image acquisition may be three-dimensional (3D), or alternatively, the imager 122 may provide a set of two-dimensional (2D) images suitable for reconstructing a three-dimensional image of the target area and / or its surrounding area. In addition, the ultrasound system 100 and / or the imager 122 may be used to detect the presence, type, and / or location associated with microbubble cavitation, as further described below. Additionally or alternatively, the system may include a cavitation detection device (e.g., a hydrophone or a suitable substitute) 124 to detect information associated with microbubble cavitation, and an administration system 126 for introducing a therapeutic agent and / or microbubbles parenterally into a patient, as further described below. The imager 122, the cavitation detection device 124, and / or the administration system 126 may be operated using the same controller 108 that facilitates operation of the transducer; alternatively, they may be separately controlled by one or more separate controllers that communicate with each other.

[0068] Figure 1BAn exemplary imager, namely an MRI device 122, is illustrated. The device 122 may include a cylindrical electromagnet 134 that generates a desired static magnetic field within a bore 136 of the electromagnet 134. During a medical procedure, a patient is placed within the bore 136 on a movable support table 138. A region of interest 140 within the patient (e.g., the patient's head) may be positioned within an imaging region 142, wherein the electromagnet 134 generates a substantially uniform field. A set of cylindrical magnetic field gradient coils 144 may also be provided within the bore 136 and around the patient. The gradient coils 144 generate magnetic field gradients of predetermined magnitude at predetermined times and in three mutually orthogonal directions. Through the field gradients, different spatial locations may be associated with different precession frequencies, thereby giving the MR image its spatial resolution. An RF transmitter coil 146 surrounding the imaging region 142 transmits RF pulses into the imaging region 142 to cause the patient's tissue to emit magnetic resonance (MR) response signals. The raw MR response signals are sensed by the RF coil 146 and passed to the MR controller 148, which then calculates an MR image that can be displayed to a user. Alternatively, separate MR transmitter and receiver coils can be used. Images acquired using the MRI device 122 can provide radiologists and physicians with visual contrast between different tissues and detailed internal views of patient anatomy that cannot be visualized with conventional x-ray techniques.

[0069] The MRI controller 148 can control the pulse sequence, i.e., the relative timing and intensity of the magnetic field gradient and the RF excitation pulses, and the response detection period. The MR response signals are amplified, conditioned, and digitized into raw data using a conventional image processing system, and further converted into an image data array by methods known to those of ordinary skill in the art. Based on the image data, a target area (e.g., a tumor) can be identified.

[0070] In order to carry out targeted drug delivery, it is necessary to determine the position of the target area with high precision before drug administration. Therefore, in various embodiments, the imager 122 is first activated to collect images of the target area and / or non-target area (e.g., healthy tissue around the target area and / or intermediate tissue between the transducer array 102 and the target area), and based on this, the anatomical features (e.g., position, size, density, structure and / or shape) associated therewith are determined. For example, the tissue volume can be represented as a 3D voxel set (i.e., volume pixels) based on a 3D image or a series of 2D image fragments, and can include a target area and / or a non-target area.

[0071] refer to Figure 2In various embodiments, after identifying a set of 3D voxels corresponding to a target volume 202 using the imager 122, the transducer array 102 is activated to generate a focal region 204 in the target volume 202. Typically, the size of the target volume 202 is larger than the size of the focal region 204. Therefore, the transducer array 102 can be sequentially activated to generate multiple focal regions 204 in the target volume 202 for damaging tissue therein, thereby temporarily increasing the permeability of the tissue. In addition, each focal region 204 can be shaped (e.g., shaped into a focal point or volume, such as a sphere or annulus) to conform to the local shape of the target region 202. Methods for configuring ultrasound transducer elements to generate focal regions of a desired size and shape are provided, for example, in U.S. Pat. No. 7,611,462, the contents of which are incorporated herein by reference.

[0072] Typically, the degree of permeability and / or size of the tissue region with increased permeability depends on the intensity and / or duration of the ultrasound application. Thus, by adjusting the ultrasound intensity and / or duration, the tissue permeability of the target area can be increased to a desired degree to allow the therapeutic agent to penetrate and / or diffuse therein and be monitored as described below. In some embodiments, the ultrasound program is adjusted based on the monitored permeability.

[0073] refer to Figure 3 In various embodiments, the ultrasonic energy emitted by the transducer element 104 may be above the threshold, resulting in a small group of bubbles (or "microbubbles") 302 in the liquid contained in the target area 202. When the heated liquid breaks and fills with gas / vapor, or when a gentle acoustic field is applied to the tissue containing the cavitation nucleus, the negative pressure generated by the propagating ultrasound or pulse can form microbubbles 302. However, at relatively low acoustic powers (e.g., 1-2 watts (Watt) above the microbubble generation threshold), the generated microbubbles 302 tend to experience oscillations and the magnitude of compression and loosening is equal, so the microbubbles usually remain unbroken (i.e., "stable cavitation"). At higher acoustic powers (e.g., more than 10 watts above the microbubble generation threshold), the generated microbubbles 302 experience loosening greater than compression, which may lead to inertial (or transient) cavitation of the microbubbles, in which the microbubbles in the liquid quickly break. Microbubble cavitation, in turn, may cause instantaneous destruction of tissue in the target area 202, thereby increasing the tissue permeability of the target area. The degree of increased permeability may depend on the microbubble concentration and / or the delivered acoustic power (or power density) and energy in the target area 202. Thus, the desired tissue permeability (i.e., allowing penetration / diffusion of therapeutic agents) may be achieved by adjusting microbubble characteristics (e.g., concentration, administration profile, etc.) and / or ultrasound parameters (e.g., amplitude, frequency, application duration, etc.), as further described below.

[0074] In some embodiments, microbubbles are injected into the target area 202 to assist in destroying tissue, thereby increasing its permeability. Microbubbles can be introduced in the form of droplets, then evaporated into bubbles filled with gas, or entrained with another suitable material (such as conventional ultrasound contrast agent). The injected microbubbles themselves may produce or promote the generation of additional microbubbles. For example, the application system 126 can introduce seed microbubbles into the target area 202, and then the controller 108 can focus ultrasound / pulses in the region near the seed microbubbles, thereby inducing the generation of microbubble groups, to destroy the tissue at the target area 202. Therefore, the actual destructive effect on the target tissue may be caused by the combination of the injected microbubbles and the microbubbles produced additionally in the tissue.

[0075] In some embodiments, ultrasound-induced microbubble cavitation is used to instantaneously destroy (or "open") the targeted BBB region. It has been found that opening the BBB reduces the burden of amyloid plaques, thereby providing therapeutic value for Alzheimer's disease. In addition, destroying the BBB region may cause the therapeutic agent in the bloodstream to penetrate the "opened" BBB region and effectively deliver therapy to targeted brain cells. Similarly, the degree and size of BBB opening can be controlled by adjusting microbubble characteristics and / or ultrasound parameters. For example, in order to effectively and efficiently cause the generation and / or cavitation of microbubbles in the target area 202, it may be necessary to maximize the amount of acoustic energy transmitted to the target area 202, while minimizing the exposure of healthy non-target tissues (e.g., tissues between the transducer and the target area) to ultrasound. Typically, the degree of absorption of ultrasound by tissues is a function of frequency, given by the following formula:

[0076] I=I0e -2αfz

[0077] Where I0 is the intensity of ultrasound when it enters the tissue (in W / cm 2 ), I is the intensity of the beam after propagating through the tissue a distance z (measured in cm), f is the frequency of the ultrasound (measured in MHz), and α is the absorption coefficient at that frequency (measured in cm- 1 MHz- 1Measured). Higher values ​​of the product α·f produce a greater degree of absorption in the target area, but a larger fraction of ultrasound is absorbed before reaching the target area. Therefore, at a certain depth z in the tissue, the ultrasound frequency of the applied wave may reflect a trade-off between acoustic power absorption in the path area and peak intensity in the focal area. In some embodiments, the optimal ultrasound transmission frequency is determined based on anatomical features (e.g., type, size, location, properties, structure, thickness, density, etc.) of the target and / or intermediate tissues to achieve peak intensity at the target area 202. Based on this, the transducer element can then be activated to cause the generation and / or cavitation of microbubbles. For example, a method for determining the optimal frequency for ultrasound application is provided in U.S. Patent Publication No. 2016 / 0008633, the contents of which are incorporated herein by reference.

[0078] In addition, if the microbubbles are preformed and introduced into the target volume 202 by the administration system 106, it may be necessary to select their size distribution so that the microbubble resonant frequency is different from the ultrasound transmission frequency to avoid damaging the non-target area. Generally, the smaller the radius of the microbubbles, the greater their resonant frequency. Therefore, once the optimal ultrasound frequency is determined, the average radius of the microbubbles having a resonant frequency substantially equal to the ultrasound frequency can be determined. In one embodiment, the size distribution of the preformed microbubbles is selected so that a large portion (e.g., more than 50%, 90%, 95% or 99% or more) of the microbubbles have a radius lower than the radius corresponding to the resonant frequency equal to the applied ultrasound frequency. Preferably, the microbubble resonant frequency is substantially greater than the ultrasound frequency (e.g., ten times greater), but if necessary, it can be substantially less than the ultrasound frequency. Therefore, when the transducer element 104 is activated with low acoustic power, the microbubbles at the non-target area do not react to the relatively low acoustic field, while the microbubbles at the target area (where the acoustic field is relatively high due to the focused beam) 202 may oscillate and / or rupture. Thus, this approach can destroy tissue at the target volume 202 with high spatial precision and avoid undesirable collateral damage to healthy tissue surrounding the target.

[0079] The ultrasound controller 108 and / or the MR controller 148 may include one or more modules implemented in hardware, software, or a combination of both. For embodiments in which the functionality is provided as one or more software programs, the programs may be written in any of a variety of high-level languages, such as PYTHON, FORTRAN, PASCAL, JAVA, C, C++, C#, BASIC, various scripting languages, and / or HTML. In addition, the software may be implemented in assembly language for a microprocessor resident on a target computer; for example, if the software is configured to run on an IBM PC or PC clone, the software may be implemented in Intel 80×86 assembly language. The software may be embodied on an article of manufacture, including, but not limited to, a floppy disk, a jump drive, a hard disk, an optical disk, a magnetic tape, a PROM, an EPROM, an EEPROM, a field programmable gate array, or a CD-ROM. Embodiments using hardware circuitry may be implemented using, for example, one or more FPGA, CPLD, or ASIC processors.

[0080] The therapeutic agent may include any drug suitable for treating a tumor. For example, to treat glioblastoma (GBM), the drug may include, for example, one or more of the following or be composed of: busulfan, thiotepa, CCNU (lomustine), BCNU (carmustine), ACNU (nimustine), temozolomide, methotrexate, topotecan, cisplatin, etoposide, irinotecan / SN-38, carboplatin, doxorubicin, vinblastine, vincristine, procarbazine, paclitaxel, tadalafil, tadalafil, succinate ... axel), Fotemustine, Ifosfamide / 4-hydroxyifosfamide / formylifosfamide, Bevacizumab, Nimotuzumab, 5-fluorouracil, Bleomycin, Hydroxyurea, Docetaxel, Cytarabine (cytosine arabinoside, ara-C) / ara-U, etc.

[0081] In addition, to treat GBM, those skilled in the art can select drugs and BBB opening schemes optimized to enhance the absorption of drugs across the BBB within patient safety limits. In this regard, it is known that the BBB in the core of many tumors has actually been destroyed, thereby allowing anti-tumor drugs to partially penetrate; but the BBB around the "brain adjacent to the tumor" (BAT) area is extensively intact, where invasive / escape GBM cells can be found and cause tumor recurrence. Ultrasound as described herein can be used to overcome the BBB, so as to better deliver drugs in the tumor core and BAT. The drugs used have different degrees of toxicity and different percentages of penetration through the BBB. The ideal drug has higher cytotoxicity to the tumor, and no BBB penetration (so that its absorption and cytotoxic effects can be limited to the area where the BBB is destroyed), lower neurotoxicity (avoiding damage to the nervous system) and tolerable systemic toxicity (e.g., below threshold) at a prescribed dose. The drug can be administered intravenously, or in some cases administered by injection near the tumor area. Additionally, configurations of administration system 106 for introducing microbubbles and / or therapeutic agents into target area 202 may be found in US Patent Application No. 62 / 597,076, the contents of which are incorporated herein by reference.

[0082] 2. Monitoring Tissue Permeability

[0083] As explained above, the observed The relationship between the parameters and the amount of injected drug reaching the target area after ultrasound-induced BBB disruption can be used to track the progress of treatment. The index parameters can be measured during ultrasound treatment, and this measurement is used to estimate the penetration of the drug into the target area without the need for a separate MR acquisition.

[0084] Specifically, Parameters are often used as safety indicators during ultrasound therapy. The parameter may also serve as an indicator of the extent of BBB disruption, thereby enabling an estimation of its permeability to drugs of a given size range. The change in the value is directly related to the total amount of deoxyhemoglobin in the tissue. Since hemoglobin is a large protein (64 kDa tetramer), it may extravasate due to hemolysis during BBB treatment without causing damage to the blood vessels. The change in the value may be related to the concentration of the macromolecule.

[0085] For ease of explanation, liposomes are used as "model molecules" herein, but it should be understood that other molecules or molecular complexes, such as quantum dots or dextran, may also or alternatively be used. Liposomes can incorporate Gd ions to be tracked and measured by MRI, and can be produced in different clinically relevant sizes. Liposomal nanoparticles based on polyethylene glycol (PEG) can be used because they have clinical relevance as nanocarriers and their metabolic decay is slow, which can stabilize their substantial concentrations after passing through the BBB. In fact, during the experiment, the substantial presence of Gd-liposomes was detected in rats a few hours after injection, and remained stable for at least one month after the BBB was closed. They can potentially last for even longer periods of time, thereby providing a useful platform for sustained drug delivery in the brain.

[0086] Generated from pre- and post-treatment MR data and R1 graphs can serve as useful and Gd-liposome concentration maps. This enables the establishment of An empirical relationship between the values ​​and the Gd-liposome concentration was established, taking into account the diffusion processes after the liposomes enter the brain parenchyma. This relationship, in turn, can serve as a valid proxy for the penetration of therapeutic agents of interest of similar size under the same ultrasound conditions. changes, liposomes of different sizes may exhibit different statistical BBB transfer kinetics. The smaller the nanoparticle, the higher its parenchymal concentration. The therapeutic region may include a size-dependent concentration plateau region where the parenchymal liposome concentration is The larger the nanoparticle, the The higher the threshold dependence.

[0087] The measurements can be performed using a conventional 1.5T MRI device during or immediately after treatment without the need for radioactive tracers, long measurement times, or strong magnetic fields, while maintaining an adequate signal-to-noise ratio. The signals can predict the passage of molecules of different sizes across the BBB, thus enabling effective trans-BBB therapeutics.

[0088] Figure 4 A representative method 400 for correlating MR signals with the penetration of liposomes across the BBB is illustrated. The analysis is performed from pre- and post-treatment volumes of and R1 per voxel calculation (step 402), and then use linear interpolation to Volume reconstruction is performed with R1 voxels (size 0.5 mm3) (step 404). This is followed by a registration step (406), which computes a registration transformation (e.g., rigid, then affine) between the pre-treatment and post-treatment T1-weighted volumes (T1W) and the pre-treatment and post-treatment T2-weighted volumes (T2W). By extracting T1 and T2* relaxation times voxel by voxel from the pre-treatment and post-treatment fast spin echo volumes with different repetition times (TR) and the pre-treatment and post-treatment gradient echo volumes with different echo time (TE) values, respectively, the T1W and T2W volumes can be obtained. and R1 values ​​(step 408). The ΔR1 values ​​can be converted to Gd-liposome concentrations (step 410) using a calibration curve, such as described below. Liposome Distribution Tracking signal, so the subpoint position (an example of which is described below) can be used Figure 7 ) from A treatment mask indicating the liposome distribution is extracted from the map (step 412). The maps of the T1- and T2-weighted volumes T1W and T2W are then registered (step 414) and used as masks, which are applied to the Gd-liposome map created in step 412 to isolate the concentration changes associated with the treatment area (step 416). To model liposome diffusion, The signal graph and the Gd-liposome concentration graph are compared to find the geometric relationship between the two parameters (step 418).

[0089] Figure 5A and Figure 5B Representative diagrams illustrate various embodiments. The relationship between the concentration of Gd-liposomes. Figure 5A A voxel-by-voxel representation of the different Gd-liposome populations in the rat model is shown. The circles 500 are the voxel-by-voxel data points, the curve 504 is the average of every 100 adjacent points, and the bars 504 are the standard deviations. The differences observed between the rats are clear. Figure 5B Shows the Figure 5A Same analysis, but pooling data for all rats in the group. Graphical plot 506 contains interpolated curves based on the three plots on the left. These plots indicate that for the same values, smaller liposomes had higher parenchymal concentrations, while larger liposomes exhibited higher Threshold dependence.

[0090] Fig. 6A A plot 600 of the particle size distribution expressed as frequency calculated from the DLS data is shown, with the mean and standard deviation for each population indicated at 602. The numbers above the curve (17.5 nm, 23.5 nm, and 43.5 nm) indicate the peaks of the distribution.

[0091] Figure 6B A graphical plot 604 of R1 versus Gd-labeled liposome concentration is shown. Calibration was performed using a series of polycarbonate wells, each containing 0.5 ml of rat blood with different Gd-liposome concentrations. To maintain a consistent R1 baseline, the rat blood content was kept the same in each liposome population measurement. Each calibration was performed on blood from one rat. Population 1 corresponds to 17 nm liposomes, population 2 corresponds to 24 nm liposomes, and population 3 corresponds to 44 nm liposomes.

[0092] As mentioned above, the subpoint position can be used from Treatment masks indicating liposome distribution are extracted in the figure. These are treatment-related The subspot can be used as a mask to isolate the liposome area, its size indicates the number of liposomes and allows calculation of the liposome concentration for a given residual volume. Figure 7 This is illustrated in A-7C. Figure 7 A indicates the position of subpoint 700 in the T2W diagram. Figure 7 B shows the treatment area The relationship between the value and the ultrasonic treatment target; specifically, Figure 7 The diameter of the circle shown in C corresponds to the lateral spot width of the ultrasound beam (e.g., 3 mm). Assuming that the liposomes undergo diffusion once they enter the brain parenchyma, Another affine registration is performed between the liposome concentration and the sub-dot map to track the diffusion. Finally, the liposome concentration can be correlated with the Plot of signal changes.

[0093] like Figure 8 A and Figure 8 As indicated in B, the R1 signal may be similarly employed. Figure 8 A depicts a graph of R1 indicating the presence of Gd in 100 nm liposomes and a graphical plot of R1 against Gd concentration in rat blood; and Figure 8 B shows similar results for 50 nm liposomes. For the graphs and figures, ΔR1 measurements were obtained using tubes filled with Gd liposomes of known concentration (tubes covering a range of concentrations) to correlate ΔR1 with liposome concentration. The value of each pixel in the image The ΔR1 value is converted into a concentration value based on the actual measurement, thus converting the pixel level The values ​​are related to the relevant liposome concentration. This produces calibration curves 804 and 806, which relate ΔR1 values ​​to liposome concentration.

[0094] 3. Methods of destroying target tissue for treatment

[0095] Methods of destroying target tissue for treatment are provided herein in various aspects. In various embodiments, the method includes ultrasonically treating a target volume to cause damage to the target tissue therein, thereby increasing its permeability; monitoring the permeability of the target tissue caused by the damage; and stopping the ultrasonic treatment of the target volume when the determined permeability reaches a threshold. In some embodiments, the ultrasound system 100 of any embodiment disclosed herein can be used to generate and deliver a focused acoustic energy beam to a target area to destroy tissue, thereby causing increased tissue permeability therein.

[0096] In some embodiments, permeability is calculated based on the measurement of a detection marker within a target tissue, where the detection marker has been administered to a subject in which permeability is being monitored.

[0097] In some embodiments, the molecule used to measure permeability is a detection marker applied to a subject. In these embodiments, the measurement of the detection marker in the target tissue indicates that the detection marker is transported from the blood to the target tissue when ultrasound is applied. In these embodiments, after inducing tissue permeability, these detection markers in the target tissue are detected and measured for the transportation of therapeutic agents with molecular weights similar to the detection markers. In these embodiments, after inducing tissue permeability, these detection markers in the target tissue are detected and measured for standardization to compare the transportation of therapeutic agents with molecular weights similar to biomarkers between different tissue permeability induction applications. In some embodiments, the detection marker is an MRI visible contrast agent. In some embodiments, the MRI visible contrast agent includes ions Gd). In some embodiments, the detection marker is a positron emission tomography (PET) reporter. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol and 18F-fluorodopa (F-Dopa). In some embodiments, the detection marker is a single photon emission computed tomography (SPECT) reporter. In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane and 99mTc-TRODAT-1). The size of these agents can be adjusted by chemical conjugation with entities such as polymers (but not limited to, for example, dextran and polyethylene glycol), or by complexing with large complexes such as liposomes or quantum dots.

[0098] In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome having a defined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is an MRI visible contrast agent, which optionally includes the ion Gd. In some embodiments, the sonication comprises at least two applications on different days in the same patient. In some embodiments, the sonication comprises different applications in different patients.

[0099] In some embodiments, permeability is based on the measurement of the level of molecules that are mainly present in the target tissue but not substantially present in the tissue outside the target tissue under normal conditions in the biological sample from the subject in which the permeability is being monitored. In some embodiments, the target tissue is the brain. In some embodiments, the biological sample is blood. In some embodiments, the molecule that is mainly present in the target tissue but not substantially present in the tissue outside the target tissue under normal conditions is a brain protein. In some embodiments, brain proteins are usually not found outside the skull tissue (e.g., in the bloodstream). In some embodiments, when the blood-brain barrier is fully opened to allow molecules with the size or molecular weight of brain proteins to leak, brain proteins will leak outside the skull tissue (e.g., in the bloodstream). In some embodiments, when the blood-brain barrier is fully opened to allow molecules with the size or molecular weight of brain proteins to leak, brain proteins become detectable in biological samples (e.g., blood samples). Illustrative, non-limiting examples of molecules that are primarily present in the target tissue but are substantially absent from tissues outside the target tissue include brain proteins, including but not limited to NSE, S100B, GFAP and UCH-L1, MBP, tau, and NfL. Murcko et al., Diagnostic biomarker kinetics: how brain-derived biomarkers distribute through the human body, and how this affects their diagnostic significance: the case of S100B. Fluids Barriers CNS 2022; 19: 32; Janigro et al., Peripheral Blood and Salivary Biomarkers of Blood-Brain Barrier Permeability and Neuronal Damage: Clinical and Applied Concepts, Front Neurol. 2021; 11: 577312. The native form of NSE is a homodimer with a molecular weight of approximately 78 kDa. See et al., Purification and characterization of human neuron-specific enolase: radioimmunoassay development, Tumour Biol 1984; 5(2): 127-39. S100B is also a homodimeric protein of 21.5 kDa, although homotetrameric and homohexameric forms are known. See Thulin et al., Molecular determinants of S100B oligomer formation, PLoS One 2011; 6(3): e14768. Thus, in some embodiments, the NSE species detected has a molecular weight of about 78 kDa. In some embodiments, the S100B species detected has a molecular weight of about 22 kDa. In some embodiments, the GFAP species detected are monomers (50 kDa), dimers (100 kDa) and / or tetramers (200 kDa). Tang et al., J Biol Chem. Oligomers of Mutant Glial Fibrillary Acidic Protein (GFAP) Inhibit the Proteasome System in Alexander Disease Astrocytes, and the Small Heat Shock Protein αB-Crystallin Reverses the Inhibition, 2010; 285(14): 10527-10537. Similarly, in some embodiments, UCH-L1, MBP, tau, and NfL species have molecular weights of about 25 kDa, about 18,500 Daltons, about 62 kDa, and about 68 kDa, respectively.

[0100] In some embodiments, the sonication comprises at least two applications on different days in the same patient. In some embodiments, the sonication comprises different applications in different patients.

[0101] In some embodiments, the molecule used to measure permeability is a biomarker specific to the target tissue. In these embodiments, the measurement of the detection marker outside the target tissue indicates that the biomarker is transported from the target tissue to the outside of the target tissue (but not limited to, for example, in the blood) when ultrasound is applied. In these embodiments, after inducing tissue permeability, these molecules outside the target tissue are detected and measured for the transportation of therapeutic agents with molecular weights similar to the molecules. In these embodiments, after inducing tissue permeability, these molecules outside the target tissue are detected and measured for standardization to compare the transportation of therapeutic agents with molecular weights similar to biomarkers between different tissue permeability induction applications. In some embodiments, target tissue-specific biomarkers include central nervous system tissue-specific markers, such as NSE and S100B, which are not usually found in the blood. In some embodiments, the detected NSE species has a molecular weight of about 78kDa. In some embodiments, the detected S100B species has a molecular weight of about 22kDa. In some embodiments, the detected NSE species has a molecular weight of about 78kDa. In some embodiments, the detected S100B species has a molecular weight of about 22 kDa. In some embodiments, the detected GFAP species is a monomer (50 kDa), a dimer (100 kDa) and / or a tetramer (200 kDa). Tang et al., J Biol Chem. Oligomers of Mutant Glial Fibrillary Acidic Protein (GFAP) Inhibit the Proteasome System in Alexander Disease Astrocytes, and the Small Heat Shock Protein α B-Crystallin Reverses the Inhibition, 2010; 285 (14): 10527-10537. Similarly, in some embodiments, UCH-L1, MBP, tau and NfL species have molecular weights of about 25 kDa, about 18,500 Daltons, about 62 kDa and about 68 kDa, respectively. Therefore, in some embodiments, the measurement of NSE species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 60 kDa to about 100 kDa. In some embodiments, the measurement of S100B species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 15 kDa to about 30 kDa. In some embodiments, the measurement of GFAP species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 40 kDa to about 250 kDa, depending on the species detected. In some embodiments, the measurement of UCH-L1 species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 20 kDa to about 30 kDa.In some embodiments, the measurement of MBP species is used to standardize the transport of therapeutic agents with a molecular weight range of about 15kDa to about 20kDa. In some embodiments, the measurement of tau species is used to standardize the transport of therapeutic agents with a molecular weight range of about 50kDa to about 70kDa. In some embodiments, the measurement of NfL species is used to standardize the transport of therapeutic agents with a molecular weight range of about 60kDa to about 80kDa. Therefore, in various embodiments, the measurement of one or more molecular species that are primarily present in the target tissue but are substantially absent in tissues outside the target tissue is used to standardize the transport of therapeutic agents with a molecular weight range of about 15kDa to about 250kDa.

[0102] 4. Methods for comparing the permeability of target tissues between at least two applications

[0103] Provided herein in various aspects are methods for comparing the permeability of a target tissue between at least two ultrasonic treatment applications. In various embodiments, the method includes providing a biological sample from a subject who has received a first ultrasonic treatment application to a target volume to cause damage to the target tissue therein, thereby increasing its permeability. In some embodiments, the target tissue is brain and / or the biological sample is blood. In some embodiments, the method also includes measuring the amount of molecules in the biological sample that are mainly present in the target tissue under normal conditions but are substantially absent from tissues outside the target tissue. In some embodiments, any technique known to the technician can be used to measure the molecule. In some embodiments, the method also includes monitoring the permeability of the target tissue caused by the damage. In some embodiments, the method also includes providing a second biological sample from a subject who has received a second ultrasonic treatment application to the target volume on a different date to cause a second damage to the target tissue therein, thereby increasing its second permeability. In some embodiments, the method also includes measuring the amount of molecules in the second biological sample that are mainly present in the target tissue under normal conditions but are substantially absent from tissues outside the target tissue. In some embodiments, the method also includes monitoring the permeability of the target tissue caused by the second damage. In some embodiments, the method further comprises comparing the permeability of the target tissue in the subject between at least two sonication applications on different days.

[0104] In some embodiments, the target tissue is brain and / or the biological sample is blood. In some embodiments, the molecule is mainly present in the target tissue under normal conditions, but is substantially absent in tissues outside the target tissue. Illustrative, non-limiting examples of molecules that are mainly present in the target tissue but are substantially absent in tissues outside the target tissue include, but are not limited to, brain proteins NSE, S100B, GFAP and UCH-L1, MBP, tau and NfL. In some embodiments, the detected NSE species has a molecular weight of about 78kDa. In some embodiments, the detected S100B species has a molecular weight of about 22kDa. In some embodiments, the detected NSE species has a molecular weight of about 78kDa. In some embodiments, the detected S100B species has a molecular weight of about 22kDa. In some embodiments, the detected GFAP species is a monomer (50kDa), a dimer (100kDa) and / or a tetramer (200kDa). Tang et al., J Biol Chem. Oligomers of Mutant Glial Fibrillary Acidic Protein (GFAP) Inhibit the Proteasome System in Alexander Disease Astrocytes, and the Small Heat Shock Protein αB-Crystallin Reverses the Inhibition, 2010; 285(14): 10527-10537. Similarly, in some embodiments, UCH-L1, MBP, tau, and NfL species have molecular weights of about 25 kDa, about 18,500 Daltons, about 62 kDa, and about 68 kDa, respectively. Thus, in some embodiments, measurement of NSE species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 60 kDa to about 100 kDa. In some embodiments, measurement of S100B species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 15 kDa to about 30 kDa. In some embodiments, the measurement of GFAP species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 40 kDa to about 250 kDa, depending on the species detected. In some embodiments, the measurement of UCH-L1 species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 20 kDa to about 30 kDa. In some embodiments, the measurement of MBP species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 15 kDa to about 20 kDa. In some embodiments, the measurement of tau species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 50 kDa to about 70 kDa.In some embodiments, measurement of NfL species is used to standardize the delivery of therapeutic agents with a molecular weight range of about 60 kDa to about 80 kDa. Thus, in various embodiments, measurement of one or more molecular species that are predominantly present in the target tissue but substantially absent from tissues outside the target tissue is used to standardize the delivery of therapeutic agents with a molecular weight range of about 15 kDa to about 250 kDa.

[0105] In some embodiments, the molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue are tumor-derived markers, such as circulating tumor cells, circulating tumor DNA, tumor extracellular vesicles, etc. In some embodiments, the molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue are tumor-derived markers, such as circulating tumor cells, circulating tumor DNA, tumor extracellular vesicles, etc. In some embodiments, the molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue are biomarkers of stroke, including but not limited to S100B, NSE, MBP and GFAP. In some embodiments, the molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue are biomarkers of neurodegenerative disorders, including but not limited to axonal proteins and NfL. In some embodiments, the molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue are biomarkers of Alzheimer's disease (AD), including but not limited to phosphorylated tau proteins P-tau181 and P-tau217. In some embodiments, molecules that are predominantly present in the target tissue under normal conditions but are substantially absent from tissues outside the target tissue are biomarkers for traumatic brain injury, including but not limited to NfL, UCH-L1, tau, S100B, and GFAP.

[0106] In some embodiments, any technique known to a skilled artisan is used to measure molecules that are primarily present in the target tissue under normal conditions but are substantially absent from tissues outside the target tissue. In some embodiments, mass spectrometry (such as, but not limited to, surface-enhanced Raman spectroscopy (SERS) and ) is used to measure molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue. In some embodiments, immunological techniques (e.g., enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), fluorescent immunoassay, and chemiluminescent immunoassay) are used to measure molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue. In some embodiments, techniques for detecting nucleic acids are used to measure molecules that are mainly present in the target tissue under normal conditions but are substantially absent in tissues outside the target tissue, including but not limited to quantitative PCR (Q-PCR), droplet digital polymerase chain reaction (ddPCR), methylation-specific PCR (MS-PCR), beads, emulsions, amplification, and magnetism (BEAMing), safe sequencing system (Safe-SeqS), cancer personalized profiling by deep sequencing (CAPP-Seq), tag amplicon deep sequencing (TAm-Seq), whole genome sequencing (WGS), or whole exome sequencing (WES).

[0107] On the other hand, a method for comparing the permeability of target tissue in a subject between at least two focused ultrasound treatment applications on different dates is provided. In some embodiments, the method includes providing a subject who has received a first ultrasound treatment application to a target volume to cause damage to the target tissue therein, thereby increasing its permeability. In some embodiments, the subject has received a detection marker. In some embodiments, the subject has received a detection marker on the same day. In some embodiments, the method also includes measuring the level of the detection marker in the target tissue, thereby monitoring the permeability of the target tissue caused by the damage. In some embodiments, the method also includes providing a second biological sample from a subject, the subject has received a second ultrasound treatment application to the target volume on a different date to cause a second destruction of the target tissue therein, thereby increasing its second permeability, optionally wherein the subject has received a detection marker on the same day. In some embodiments, the method also includes measuring the level of the detection marker in the target tissue, thereby monitoring the permeability of the target tissue caused by the second destruction, thereby monitoring the permeability of the target tissue caused by the second destruction. In some embodiments, the method also includes comparing the permeability of the target tissue in the subject between at least two ultrasound treatment applications on different dates.

[0108] In some embodiments, the target tissue is the brain. In some embodiments, the target tissue is the brain, and the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent. In some embodiments, the target tissue is the brain, and the detection marker is complexed with a liposome, which has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is an MRI visible contrast agent. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the detection marker is a positron emission tomography (PET) reporter. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol and 18F-fluorodopa (F-Dopa). In some embodiments, the detection marker is a single photon emission computed tomography (SPECT) reporter. In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1).

[0109] It should be understood that calibration can be achieved in any conventional manner and may involve machine learning techniques, linear or nonlinear regression, or other suitable techniques. In addition, after calibration, the controller 108 can be configured or programmed to dynamically control ultrasound therapy in response to ongoing permeability estimates.

[0110] It was found that, unlike low-weight Gd-based contrast agents that are completely washed out of the parenchyma a few hours after administration, liposomes can be detected in the brain a few hours after injection and stay in the tissue for at least one month after administration. This phenomenon can be explained by the size of the liposomes, which are an order of magnitude larger than the Gd salts, thus complicating their transfer through the BBB; and the long half-life of PEG-based liposomes in the blood (tens of hours), which causes the liposomes to accumulate in the parenchyma and slow down the washing process as the BBB closes, until its parenchymal concentration eventually stabilizes. Therefore, the liposome concentration can be determined at least in part by the perfusion time or the washing time. Specifically, since the molecules quickly cross the BBB, a high perfusion time-washing time corresponds to a high permeability.

[0111] The terms and expressions used herein are used as terms and expressions of description rather than limitation, and when such terms and expressions are used, it is not intended to exclude any equivalents of the features shown and described or parts thereof. In addition, although certain embodiments of the present invention have been described, it will be apparent to those of ordinary skill in the art that other embodiments incorporating the concepts disclosed herein may be used without departing from the spirit and scope of the present invention. Therefore, the described embodiments should be considered in all respects to be illustrative only and not restrictive.

[0112] 5. Neurological diseases or conditions that can be treated using the methods and systems disclosed herein

[0113] In some aspects, the disclosure provides methods for treating a neurological disease or disorder in a subject in need thereof, wherein the neurological disease or disorder is characterized by abnormal production, aggregation and / or deposition of proteins or other biomolecules in the brain. In some embodiments, the neurological disease or disorder is selected from Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), dementia with Lewy bodies, spinocerebellar ataxia and amyotrophic lateral sclerosis, frontotemporal lobar disease, multiple system atrophy, four-repeat tauopathy and prion disease.

[0114] Alzheimer's disease patients exhibit senile plaques composed mainly of amyloid-β (Aβ), neurofibrillary tangles including Tau protein, neuronal inclusions of TDP-43, and Lewy bodies including α-synuclein. Parkinson's disease patients exhibit Lewy bodies including α-synuclein. Patients with amyotrophic lateral sclerosis have neuronal inclusions, which include TAR DNA binding protein 43 (TDP-43), sarcoma fusion / liposarcoma transporter (FUS / TLS) and superoxide dismutase-1 (SOD1). Huntington's disease is a progressive brain disorder caused by mutations in the gene encoding huntingtin protein, resulting in the production of abnormal mutant proteins that gradually damage brain cells. Lewy body dementia is characterized by Lewy bodies including α-synuclein, senile plaques composed mainly of amyloid-β (Aβ) and neurofibrillary tangles of Tau protein. Patients with frontotemporal lobe diseases exhibit neuronal and glial inclusions composed of Tau, TDP-43 and FUS / TLS. Multiple system atrophy is characterized by glial cytoplasmic inclusions of α-synuclein. Therefore, there seems to be overlap between the proteins that exhibit abnormal production, aggregation and / or deposition associated with these diseases. A single neurodegenerative disease may be associated with a variety of proteins (or another biomolecule) that exhibit abnormal production, aggregation and / or deposition. On the other hand, a single protein that exhibits abnormal production, aggregation and / or deposition may also be associated with a variety of diseases. For example, although Aβ plaques and tau tangles are typical features of Alzheimer's disease, Lewy bodies typical of Parkinson's disease are found in more than 50% of Alzheimer's disease cases, and neuronal inclusions composed of protein TDP-43 are found in more than 40% of Alzheimer's disease cases. Similarly, in dementia with Lewy bodies, a dementia closely related to Parkinson's disease that shares some features of Alzheimer's disease, the typical α-synuclein-rich Lewy bodies are associated with Aβ plaques in 60% of cases and tau tangles in 50% of cases. Similarly, the quadruple-repeat tauopathies are a group of neurodegenerative diseases defined by cytoplasmic inclusions composed primarily of a tau isoform with four microtubule-binding domains that are associated with at least three clinical manifestations: (1) progressive supranuclear palsy presenting with axial rigidity and oculomotor problems in addition to atypical Parkinsonism; (2) corticobasal degeneration presenting as frontal lobe dementia with focal cortical syndromes, including progressive apraxia or progressive aphasia; and (3) argyrophilic grain disease, an increasingly recognized geriatric disorder that affects the medial temporal lobe and is associated with amnestic cognitive impairment.

[0115] Thus, in some aspects, the present disclosure provides a method of treating a neurological disease or disorder in a subject in need thereof, wherein the neurological disease or disorder is characterized by abnormal production, aggregation and / or deposition of a protein or another biomolecule in the brain, wherein a therapeutic agent and / or a microbubble composition is to be, is being, or has been administered to the subject, the method comprising the steps of: sonicating a target volume to cause damage to a target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and an adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the damage; and when the permeability reaches a threshold value, ceasing sonication of the target volume, thereby increasing the level of delivery of the therapeutic agent to the site compared to a control.

[0116] In another aspect, the present invention relates to a method for treating a neurological disease or disorder in a subject in need thereof, wherein the neurological disease or disorder is characterized by a site in the brain having abnormal production, aggregation and / or deposition of a protein or another biomolecule, the method providing a predetermined range of local doses of a therapeutic agent, the method comprising the following steps: (i) subjecting the target volume to ultrasound treatment to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and the adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, microbubble composition and / or detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue, thereby monitoring the local dose of the therapeutic agent caused by the damage at the site; (iii) stopping ultrasound treatment of the target volume when the local dose of the therapeutic agent reaches a predetermined range. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome having a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0117] In some embodiments, the control is the delivery level of the therapeutic agent in the subject who has not yet received the acoustic pulse sequence and / or microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before the acoustic pulse sequence and / or microbubble composition is applied. In some embodiments, the nervous system disease or disorder is Alzheimer's disease, and the position is selected from the senile plaques comprising amyloid-β (Aβ), the neurofibrillary tangles comprising Tau protein, the neuronal inclusions comprising TDP-43 and the Lewy bodies comprising α-synapse nucleoprotein. In some embodiments, the nervous system disease or disorder is Parkinson's disease, and the position is the Lewy bodies comprising α-synapse nucleoprotein. In some embodiments, the nervous system disease or disorder is amyotrophic lateral sclerosis, and the position is the neuronal inclusions comprising TARDNA binding protein 43 (TDP-43), sarcoma fusion / liposarcoma transporter (FUS / TLS) and superoxide dismutase-1 (SOD1). In some embodiments, the nervous system disease or disorder is Huntington's disease, and the site is the neuronal nuclear inclusions of Huntington protein. In some embodiments, the nervous system disease or disorder is Lewy body dementia, and the site is Lewy bodies containing α-synuclein, senile plaques containing amyloid-β (Aβ), and neurofibrillary tangles containing Tau protein. In some embodiments, the nervous system disease or disorder is frontotemporal lobe disease, and the site is neurons and glial inclusions composed of Tau, TDP-43 and FUS / TLS. In some embodiments, the nervous system disease or disorder is multiple system atrophy, and the site is the glial cytoplasmic inclusions of α-synuclein. In some embodiments, the nervous system disease or disorder is four-repeat tauopathy, and the site is a cytoplasmic inclusion composed mainly of tau protein subtypes with four microtubule binding domains.

[0118] Diseases associated with aggregation and / or accumulation of a protein (or another biomolecule) that exhibits abnormal production, aggregation, and / or deposition also include prion diseases, i.e., transmissible spongiform encephalopathies, such as bovine spongiform encephalopathy (BSE or mad cow disease) and Creutzfeldt-Jakob disease. These diseases are characterized by senile plaques composed of PrP protein. Thus, in some aspects, the present disclosure provides a method for treating prion disease, the method comprising: (a) selecting a subject having a PrP protein deposition site in the brain; (b) identifying a blood-brain barrier (BBB) ​​region for ultrasound treatment, wherein the region is adjacent to or completely encompasses the site; (c) administering a therapeutic agent to the subject; and (d) applying an ultrasound beam through the subject's skull to the BBB region to facilitate transport of the therapeutic agent across the BBB to the site.

[0119] The present disclosure provides a method for treating prion disease, which provides a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a magnetic resonance imaging (MRI) visible contrast agent, a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent comprises ion Gd. In some embodiments, the PET reporter is selected from 18 F-Flubetapyr, 18 F-Fluorobetaben, 18 F-Flutociclib, 18 F-Flumetazol and 18 F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99m Tc-ECD (ethylcysteine ​​dimer), 123 I-iodoflurane and 99m Tc-TRODAT-1.

[0120] Alzheimer's disease

[0121] Alzheimer's disease (AD) is a complex, progressively debilitating, and fatal neurodegenerative disease. The frequency of AD is increasing rapidly as the world's population ages. Currently, an estimated 6.5 million people in the United States suffer from AD, a number that is expected to increase to more than 13 million by 2050. Approximately 15% of the U.S. population over the age of 60 has prodromal AD, and approximately 40% has preclinical AD. Similar trends are seen globally, and unless methods are found to delay, prevent, or treat AD, the worldwide AD dementia patient population is expected to exceed 100 million by 2050. There is a great need for therapeutic agents that stop or reverse the underlying pathology of AD.

[0122] The cellular and molecular mechanisms of AD are not well understood. Researchers have reported that AD is associated with genetic and environmental factors as well as lifestyle. The heterogeneity of AD patients is that they may be in the preclinical AD continuum for up to 20 years or more without showing any clinical symptoms, i.e., mild cognitive impairment (MCI), AD dementia, or functional decline. In addition, misdiagnosis of AD patients is also common, as 10%-30% of individuals clinically diagnosed with AD dementia do not show AD neurodegeneration at autopsy.

[0123] Among all types of AD therapies, the failure rate exceeds 99%, while for disease modifying therapies (DMTs), the failure rate is 100%. Therefore, in addition to new methods for developing therapeutic agents, there is a need for methods for targeted delivery of therapeutic agents, such as those disclosed herein.

[0124] Alzheimer's disease is associated with senile plaques composed of amyloid-β (Aβ), neurofibrillary tangles including Tau protein, neuronal inclusions of TDP-43, and Lewy bodies including α-synuclein. Aβ is a relatively small peptide of 4kDa to 4.4kDa, which is the main component of amyloid deposits. Intracellular Aβ protein is widely present in neurons, and it is associated with inflammatory and antioxidant activity, regulation of cholesterol transport, and activation of kinases. However, Aβ is one of the most well-known components in the formation of neurodegenerative diseases including AD. Aβ is composed of approximately 36-43 amino acids, and it is derived from amyloid precursor protein (APP), a glycoprotein of 695-770 amino acids. APP can be cleaved into multiple fragments by α-secretase, β-secretase, and γ-secretase, and Aβ protein is formed by the action of β-secretase and γ-secretase. Aβ protein contains two important regions, which play a major role in the formation of insoluble amyloid fibrils.

[0125] Microtubule-associated Tau protein is highly expressed in the brain, and its name is derived from "tubulin-associated units". Microtubules are the main proteins of the cytoskeleton. The main function of Tau protein is to stabilize microtubules by binding to microtubules and other proteins. In order to perform these functions, Tau protein is phosphorylated at normal levels. Hyperphosphorylation of Tau protein is believed to cause conformational changes and aggregation of tau protein. Other post-translational modifications such as glycosylation, glycation, polyamination and nitration may play a role in aggregation. Therefore, in some aspects, the present disclosure provides a method for treating Alzheimer's disease (AD), wherein a therapeutic agent and / or a microbubble composition will be, is being or has been administered to a subject, the method comprising the following steps: ultrasonically treating a target volume to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the damage; and when the permeability reaches a threshold, stopping the ultrasonic treatment of the target volume, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in a subject that has not yet received the acoustic pulse sequence and / or microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in a subject before the administration of the acoustic pulse sequence and / or microbubble composition.

[0126] In some aspects, the present disclosure provides a method for treating Alzheimer's disease (AD), which provides a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to a target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and an adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0127] In some embodiments, the protein exhibiting abnormal production, aggregation and / or deposition is selected from amyloid-β peptide (Aβ), neurofibrillary tangles and tau protein. In some embodiments, the protein exhibiting abnormal production, aggregation and / or deposition is selected from amyloid-β peptide (Aβ), TDP-43, α-synuclein and tau protein. In some embodiments, the imaging technology of a reporter capable of binding to amyloid-β peptide (Aβ), neurofibrillary tangles and / or tau protein is used to detect the site. In some embodiments, the imaging technology is PET. In some embodiments, positioning includes administering a PET reporter to a subject. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin and 18F-flumetazol. Additionally or alternatively, in some embodiments, the imaging technology is SPECT. In some embodiments, positioning includes administering a SPECT reporter to a subject. In some embodiments, the SPECT reporter is 99mTc-ECD (ethylcysteine ​​dimer). In some embodiments, the therapeutic agent is selected from a nonspecific clearance antibody (e.g., intravenous immunoglobulin, also known as IVIg), an anti-amyloid-β antibody (e.g., aducanumab, gantuzumab, lencanezumab, and donenetumab), an anti-tau antibody (e.g., cirenezumab, gonelizumab, tipranezumab, and zagteneizumab), an anti-TREM2 antibody (e.g., AL002), donepezil, rivastigmine, memantine, and galantamine, and combinations thereof. In some embodiments, the therapeutic agent is aducanumab. Some therapeutic agents are disclosed in WO 2014 / 089500 and WO 2021 / 108861, the contents of which are incorporated herein by reference.

[0128] Parkinson's disease

[0129] Parkinson's disease (PD) is a long-term degenerative disorder of the central nervous system that causes unexpected or uncontrolled movements, such as tremors, stiffness, and difficulty with balance and coordination. Symptoms usually develop gradually and worsen over time. As the disease progresses, people may have difficulty walking and speaking. They may also experience mental and behavioral changes, sleep problems, depression, memory difficulties, and fatigue. The occurrence of the disease is characterized by the accumulation of misfolded alpha-synuclein in the brain. Typically, anxiety, tremors, stiffness, depression, slow movements, and abnormal postures are the most common symptoms of Parkinson's disease.

[0130] Lewy bodies (LBs), composed mainly of α-syn, are a neuropathological hallmark of Parkinson's disease (PD) patients. However, it is increasingly recognized that PD is often associated with cognitive deficits, and a significant proportion of patients eventually develop dementia.

[0131] α-Synuclein is associated with a variety of neurodegenerative diseases known as "synucleinopathies". Native unfolded α-synuclein (α-Syn) is a highly conserved protein of 14 kDa that can be localized to different areas of the brain. Since the protein shows synaptic and nuclear localization, its name is preferably "α-synuclein". α-Syn regulates dopamine neurotransmission by regulating vesicular dopamine storage. It interacts with tubulin and may function like tau protein. In addition, α-Syn shows chaperone activity in the folding of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins. α-Syn plays a vital role in PD because α-Syn is the main fibrillar component of Lewy bodies. Two mutations in the α-Syn gene, A53T and A30P, and overexpression of wild-type α-Syn increase the misfolding process and aggregation. In addition, the accumulation of abnormal forms of α-Syn inhibits proteasome function. In PD brain, α-Syn was found to be phosphorylated at Ser87 and Ser129 in aggregates. These serine residues are phosphorylated by casein kinase 1 (CK1) and casein kinase 2 (CK2). This post-translational modification is thought to have a pathological role in the fibril formation of α-Syn.

[0132] Therefore, in some aspects, the present disclosure provides a method for treating Parkinson's disease (PD), wherein a therapeutic agent and / or a microbubble composition will be, is being or has been applied to a subject, the method comprising the following steps: the target volume is ultrasonically treated to cause the destruction of the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and the permeability of the target tissue caused by the destruction is monitored; and when the permeability reaches a threshold value, the ultrasonic treatment of the target volume is stopped, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in the subject who has not yet received the acoustic pulse sequence and / or microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before applying the acoustic pulse sequence and / or microbubble composition.

[0133] In some aspects, the present disclosure provides a method for treating Parkinson's disease (PD), which provides a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0134] In some embodiments, the protein that exhibits abnormal production, aggregation and / or deposition is alpha-synuclein. In some embodiments, the imaging technology of a reporter that can bind to alpha-synuclein is used to detect the site. In some embodiments, the imaging technology is PET. In some embodiments, positioning includes administering a PET reporter to the subject. In some embodiments, the PET reporter is 18F-fluorodopa (F-Dopa). Additionally or alternatively, in some embodiments, the imaging technology is SPECT. In some embodiments, positioning includes administering a SPECT reporter to the subject. In some embodiments, the SPECT reporter is 123I-iodoflurane or 99mTc-TRODAT-1. In some embodiments, the therapeutic agent is selected from anti-α-synuclein antibodies (e.g., simpanezumab, putinuzumab, Lu AF82422, ABBV-0805, and MEDI1341), carbidopa-levodopa, selegiline, rasagiline, safinamide, entacapone, benztropine, tolcapone, opicapone, nuprizide, istradefylline, and amantadine, and combinations thereof.

[0135] Multiple system atrophy is characterized by glial cytoplasmic inclusions of alpha-synuclein. Therefore, in some aspects, the present disclosure provides a method for treating multiple system atrophy, wherein a therapeutic agent and / or a microbubble composition will be, is being, or has been applied to a subject, the method comprising the following steps: the target volume is sonicated to cause destruction of the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the destruction; and when the permeability reaches a threshold value, the sonication of the target volume is stopped, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in a subject who has not yet received an acoustic pulse sequence and / or a microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before applying an acoustic pulse sequence and / or a microbubble composition.

[0136] In some aspects, the present disclosure provides a method for treating multiple system atrophy, the method comprising: the method provides a predetermined range of local doses of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to a target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0137] In some embodiments, the protein that exhibits abnormal production, aggregation and / or deposition is alpha-synuclein. In some embodiments, the imaging technology of a reporter that can bind to alpha-synuclein is used to detect the site. In some embodiments, the imaging technology is PET. In some embodiments, positioning includes administering a PET reporter to the subject. In some embodiments, the PET reporter is 18F-fluorodopa (F-Dopa). Additionally or alternatively, in some embodiments, the imaging technology is SPECT. In some embodiments, positioning includes administering a SPECT reporter to the subject. In some embodiments, the SPECT reporter is 123I-iodoflurane or 99mTc-TRODAT-1. In some embodiments, the therapeutic agent is selected from anti-α-synuclein antibodies (e.g., simpanezumab, putinuzumab, Lu AF82422, ABBV-0805, and MEDI1341), carbidopa-levodopa, selegiline, rasagiline, safinamide, entacapone, benztropine, tolcapone, opicapone, nuprizide, istradefylline, and amantadine, and combinations thereof.

[0138] Lewy body dementia

[0139] Dementia with Lewy bodies is characterized by Lewy bodies consisting of alpha-synuclein, senile plaques composed primarily of amyloid-beta (Aβ), and neurofibrillary tangles of the protein tau. Dementia with Lewy bodies (DLB) is a type of progressive dementia that causes a decline in thinking, reasoning, and independent function. Its features may include spontaneous changes in attention and alertness, recurring visual hallucinations, REM sleep behavior disorder, and slow movements, tremors, or rigidity. Mutations in genes called SNCA and SNCB can cause dementia with Lewy bodies. Mutations in another gene called GBA or a version of a gene called APOE increase the risk of developing the condition but are not a direct cause. Therefore, in some aspects, the present disclosure provides a method for treating Lewy body dementia, wherein a therapeutic agent and / or a microbubble composition will be, is being, or has been applied to a subject, the method comprising the following steps: the target volume is ultrasonically treated to cause destruction of the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and the permeability of the target tissue caused by the destruction is monitored; and when the permeability reaches a threshold value, the ultrasonic treatment of the target volume is stopped, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in a subject who has not yet received an acoustic pulse sequence and / or a microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before applying an acoustic pulse sequence and / or a microbubble composition.

[0140] In some aspects, the present disclosure provides a method for treating Lewy body dementia, which provides a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0141] In some embodiments, the protein that exhibits abnormal production, aggregation and / or deposition is alpha-synuclein. In some embodiments, the imaging technology of a reporter that can bind to alpha-synuclein and / or amyloid-β (Aβ) is used to detect the site. In some embodiments, the imaging technology is PET. In some embodiments, positioning includes administering a PET reporter to a subject. In some embodiments, the PET reporter is 18F-fluorodopa (F-Dopa), 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin and 18F-flumetazol. Additionally or alternatively, in some embodiments, the imaging technology is SPECT. In some embodiments, positioning includes administering a SPECT reporter to a subject. In some embodiments, the SPECT reporter is 123I-iodoflurane or 99mTc-TRODAT-1. In some embodiments, the therapeutic agent is selected from anti-α-synuclein antibodies (e.g., simpanezumab, putizumab, Lu AF82422, ABBV-0805, and MEDI1341), rivastigmine, donepezil, galantamine, memantine, carbidopa-levodopa, and combinations thereof.

[0142] Huntington's disease

[0143] Huntington's disease (HD) is an inherited neurodegenerative disorder and the disease is caused by autosomal dominant inheritance. HD patients show involuntary muscle contractions, movement and mental disorders. The disease is autosomal dominant and affects the brain and nervous system. Huntington protein undergoes conformational changes with mutations and it shows a tendency to aggregate.

[0144] In HD, neuropathology is characterized by the accumulation of Htt protein aggregates. HD is caused by a large number of CAG repeats in the gene. It is believed that CAG repeats (polyQ) are the most important promoter of Htt protein aggregate toxicity. The polyQ region starts at residue 18, and the number of glutamine residues is the most important marker in HD. Surprisingly, 40 or more CAG repeats always produce neuropathy, while 35 or fewer CAG repeats never produce neuropathy. However, in childhood, 27 to 35 CAG repeats may develop neuropathy. Thus, in some aspects, the present disclosure provides a method for treating Huntington's disease (HD), wherein a therapeutic agent and / or a microbubble composition will be, is being, or has been administered to a subject, the method comprising the following steps: ultrasonically treating a target volume to cause damage to a target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and an adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the damage; and when the permeability reaches a threshold, stopping ultrasonic treatment of the target volume, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to a control. In some embodiments, the control is the delivery level of the therapeutic agent in a subject who has not yet received an acoustic pulse sequence and / or a microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in a subject before the application of an acoustic pulse sequence and / or a microbubble composition.

[0145] In some aspects, the present disclosure provides a method for treating Huntington's disease (HD), which provides a predetermined range of local doses of a therapeutic agent, the method comprising the following steps: (i) sonicating a target volume to cause damage to a target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and an adjacent blood-brain barrier (BBB), wherein the subject has received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker, optionally on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the sonication of the target volume. In some embodiments, the detection marker is: conjugated to a polymer (e.g., dextran or polyethylene glycol) within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or complexed with a liposome having a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0146] In some embodiments, the protein that exhibits abnormal production, aggregation and / or deposition is huntingtin. In some embodiments, the site is detected using an imaging technique that can bind to a reporter of mutant huntingtin or polyglutamine or metabolic changes associated with Huntington's disease. In some embodiments, the imaging technique is PET. In some embodiments, localization comprises administering a PET reporter to the subject. In some embodiments, the PET reporter is selected from 11C-CHDI-180R, 11C-CHDI-626, 18F-CPFPX, 11C-KF18446, 18F-MK-9470, 11C-SCH-23390, 11C-NNC-112, 11C-Raclopride, 11C-Flumazenil, 11C-ABP-688, 11C-PK11195, 18F-PBR06, 18F-JNJ42249152, 18F-MNI-659, 11C-IMA-107 and 11C-UCB-J. See Cybulska et al., Huntington's Disease: A Review of the Known PET Imaging Biomarkers and Targeting Radiotracers, Molecules 2020; 25(3), 482. Additionally or alternatively, in some embodiments, the imaging technique is SPECT. In some embodiments, localization comprises administering a SPECT reporter to the subject. In some embodiments, the SPECT reporter is 99m Tc-HMPAO. In some embodiments, the therapeutic agent is selected from an anti-huntingtin antibody and an anti-SEMA4D antibody (eg, Pepinemab).

[0147] Amyotrophic lateral sclerosis (ALS)

[0148] Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by degeneration of upper and lower motor neurons, leading to progressive paralysis of limb muscles, speech, swallowing and breathing. Patients with amyotrophic lateral sclerosis have neuronal inclusions, which include TAR DNA binding protein 43 (TDP-43), sarcoma fusion / liposarcoma transporter (FUS / TLS) and superoxide dismutase-1 (SOD1). It is believed that ALS pathology begins in a single lesion or multiple lesion sites and spreads through the neuraxis in a spatiotemporal manner. It is reported that insoluble TDP-43 in the diseased brain induces TDP-43 pathology in neuroblastoma cells that overexpress wtTDP-43, as detected by TDP-43 hyperphosphorylation, ubiquitination and aggregation. Therefore, in some aspects, the present disclosure provides a method for treating amyotrophic lateral sclerosis (ALS), wherein a therapeutic agent and / or a microbubble composition will be, is being, or has been administered to a subject, the method comprising the following steps: ultrasonic treatment of the target volume to cause destruction of the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the destruction; and when the permeability reaches a threshold value, stopping the ultrasonic treatment of the target volume, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in a subject who has not yet received an acoustic pulse sequence and / or a microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before applying an acoustic pulse sequence and / or a microbubble composition.

[0149] In some aspects, the present disclosure provides a method for treating amyotrophic lateral sclerosis (ALS), which provides a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is: conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0150] In some embodiments, the protein that exhibits abnormal production, aggregation and / or deposition is alpha-synuclein. In some embodiments, the site is detected using an imaging technique that is capable of binding to TAR DNA binding protein 43 (TDP-43), fusion sarcoma / liposarcoma transporter (FUS / TLS), and superoxide dismutase-1 (SOD1). In some embodiments, the imaging technique is PET. In some embodiments, positioning comprises administering a PET reporter to a subject. In some embodiments, the PET reporter is 18F-fluorodeoxyglucose, 18F-fluorobetaben, 11C-flumazenil, and / or 3H-APN-1607. Additionally or alternatively, in some embodiments, the imaging technique is SPECT. In some embodiments, positioning comprises administering a SPECT reporter to a subject. In some embodiments, the SPECT reporter is 99m Tc-ECD. In some embodiments, the therapeutic agent is selected from anti-TDP-43 antibody, anti-SOD1 antibody, riluzole, edaravone, sodium phenylbutyrate and taurine diol, or a combination thereof.

[0151] Spinocerebellar ataxia

[0152] Spinocerebellar ataxia (SCA) is a group of complex neurodegenerative disorders characterized by progressive cerebellar ataxia of gait and limbs variably associated with ophthalmoplegia, pyramidal and extrapyramidal signs, dementia, pigmentary retinopathy and peripheral neuropathy. The age of onset of the disease is usually between 30 and 50 years old, but there are reports of early childhood onset and decades after the age of 60. Depending on the underlying cause of the spinocerebellar ataxia subtype, the prognosis is variable. Mutations in ATXN1, ATXN2, ATXN3, SCA4, SPTBN2, CACNAIA, ATXN7, KLHL1AS, ATXN10, SCA11, PPP2R2B, KCNC3, PRKCG, etc. have been found in SCA. In addition, seven subtypes of spinocerebellar ataxias (including SCA 1, 2, 3 / Machado-Joseph disease, 6, 7, 17, and dentatorubral pallidum-Lewy body atrophy (DRPLA)) are caused by expansions of CAG repeats in specific genes, resulting in abnormally long polyQ tracts in the encoded proteins. Proteins with expanded polyglutamine stretches appear to assume abnormal conformations, leading to the formation and deposition of polyglutamine aggregates in disease neurons, forming characteristic nuclear or cytoplasmic inclusions that are the neuropathological hallmarks of these diseases. Therefore, in some aspects, the present disclosure provides a method for treating spinocerebellar ataxia, wherein a therapeutic agent and / or a microbubble composition will be, is being or has been applied to a subject, the method comprising the following steps: the target volume is ultrasonically treated to cause the destruction of the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and the permeability of the target tissue caused by the destruction is monitored; and when the permeability reaches a threshold value, the ultrasonic treatment of the target volume is stopped, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in the subject who has not yet received an acoustic pulse sequence and / or a microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before applying an acoustic pulse sequence and / or a microbubble composition.

[0153] In some aspects, the present disclosure provides a method for treating spinocerebellar ataxia, which provides a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to a target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and an adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0154] In some embodiments, the protein exhibiting abnormal production, aggregation and / or deposition is a mutant protein with an expanded polyglutamine tract. In some embodiments, the site is detected using an imaging technique that uses a reporter that can bind to an indicator of a metabolic defect associated with a poly Q tract or spinocerebellar ataxia. In some embodiments, the imaging technique is PET. In some embodiments, localization comprises administering a PET reporter to a subject. In some embodiments, the PET reporter is 18 F-fluorodeoxyglucose and / or 18 F-SDM-8. Additionally or alternatively, in some embodiments, the imaging technique is SPECT. In some embodiments, localization comprises administering a SPECT reporter to the subject. In some embodiments, the SPECT reporter is 99m Tc-TRODAT-1. In some embodiments, the therapeutic agent is selected from anti-polyglutamine antibodies.

[0155] Frontotemporal lobe disorders

[0156] The clinical syndrome of frontotemporal dementia is clinically and neuropathologically heterogeneous, but processes such as neuroinflammation may be common across the disease spectrum. In recent years, attention has focused on understanding the pathogenic role of protein misfolding and aggregation, which are the main features of the postmortem diagnostic criteria for frontotemporal lobar degeneration (FTLD). These diseases are associated with neuronal and glial inclusions composed of tau, TDP-43, and FUS / TLS. Frontotemporal dementia with parkinsonism-17 (FTDP-17) is a progressive neurodegenerative disease caused by mutations in the tau gene. The tau gene is mutated in familial FTDP-17, and this mutation accelerates the formation of neurofibrillary tangles (NFTs) in the brain. In addition, this mutation promotes hyperphosphorylation. In some aspects, the present disclosure provides a method for treating frontotemporal dementia, the method comprising: wherein to, being or having applied a therapeutic agent and / or a microbubble composition to a subject, the method comprising the following steps: ultrasonic treatment of the target volume to cause destruction of the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the destruction; and when the permeability reaches a threshold value, the ultrasonic treatment of the target volume is stopped, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in a subject who has not yet received an acoustic pulse sequence and / or a microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before applying an acoustic pulse sequence and / or a microbubble composition.

[0157] In some aspects, the present disclosure provides a method for treating frontotemporal dementia, which provides a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) that is within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome that has a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0158] In some embodiments, the site is a neuronal and glial inclusion composed of Tau, TDP-43, and FUS / TLS. In some embodiments, the proteins that exhibit abnormal production, aggregation, and / or deposition are Tau, TDP-43, and FUS / TLS. In some embodiments, the site is detected using an imaging technique that is a reporter that can bind to Tau, TDP-43, and FUS / TLS. In some embodiments, the imaging technique is PET. In some embodiments, positioning includes administering a PET reporter to a subject. In some embodiments, the PET reporter is selected from 18F-PI-2620, 18F-fluorodeoxyglucose, and / or 18F-GE-180. Additionally or alternatively, in some embodiments, the imaging technique is SPECT. In some embodiments, positioning includes administering a SPECT reporter to a subject. In some embodiments, the SPECT reporter is 99m Tc-HMPAO. In some embodiments, the therapeutic agent is an anti-tau antibody.

[0159] Quadruple repeat tauopathy

[0160] Four-repeat (4R-) tauopathies are a group of neurodegenerative diseases defined by cytoplasmic inclusions of tau protein subtypes. Progressive supranuclear palsy, corticobasal degeneration, argyrophilic grain disease or glial globular tauopathies belong to the 4R-tauopathies group. Tau is a microtubule-associated protein that has multiple functions in the dynamic assembly of the neuronal cytoskeleton, and in these diseases, cytoplasmic inclusions consisting mainly of tau protein subtypes with four microtubule binding domains have been found. In addition, the tau protein is usually located in the axons, but in tauopathies, it is located in the dendrites. Therefore, the transport system of neurons may disintegrate and microtubules may not work properly. Therefore, in some aspects, the present disclosure provides a method for treating four repeat (4R-) tauopathy, wherein a therapeutic agent and / or a microbubble composition will be, is being or has been applied to a subject, the method comprising the following steps: the target volume is ultrasonically treated to cause the destruction of the target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and the adjacent blood-brain barrier (BBB); and the permeability of the target tissue caused by the destruction is monitored; and when the permeability reaches a threshold value, the ultrasonic treatment of the target volume is stopped, thereby increasing the delivery level of the therapeutic agent delivered to the site compared to the control. In some embodiments, the control is the delivery level of the therapeutic agent in the subject who has not yet received an acoustic pulse sequence and / or a microbubble composition. In other embodiments, the control is the delivery level of the therapeutic agent in the subject before applying an acoustic pulse sequence and / or a microbubble composition.

[0161] In some aspects, the present disclosure provides a method for treating four-repeat (4R-) tauopathy, the method providing a local dose of a predetermined range of a therapeutic agent, the method comprising the following steps: (i) ultrasonically treating a target volume to cause damage to a target tissue therein, thereby increasing its permeability, wherein the target volume covers the site and an adjacent blood-brain barrier (BBB), wherein the subject has optionally received, will receive, or is receiving a dose of a therapeutic agent, a microbubble composition, and / or a detection marker on the same day; (ii) measuring the level of the detection marker in the target tissue to monitor the local dose of the therapeutic agent caused by the damage at the site; (iii) when the local dose of the therapeutic agent reaches a predetermined range, stopping the ultrasonic treatment of the target volume. In some embodiments, the detection marker is conjugated to a polymer (e.g., dextran or polyethylene glycol) within a determined molecular weight range corresponding to the molecular weight range of the therapeutic agent; or is complexed with a liposome having a determined size range corresponding to the size range of the therapeutic agent. In some embodiments, the detection marker is selected from a contrast agent visible to magnetic resonance imaging (MRI), a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter. In some embodiments, the MRI visible contrast agent includes ion Gd. In some embodiments, the PET reporter is selected from 18F-flubetapyr, 18F-flubetaben, 18F-flutocillin, 18F-flumetazol, and 18F-fluorodopa (F-Dopa). In some embodiments, the SPECT reporter is selected from 99mTc-ECD (ethylcysteine ​​dimer), 123I-iodoflurane, and 99mTc-TRODAT-1.

[0162] In some embodiments, the protein that exhibits abnormal production, aggregation and / or deposition is Tau protein. In some embodiments, the site is detected using an imaging technique that uses a reporter that can bind to Tau protein. In some embodiments, the imaging technique is PET. In some embodiments, the localization includes administering a PET reporter to the subject. In some embodiments, the PET reporter is selected from 18 F-PI-2620, 18 F-fluorodeoxyglucose and / or 18 F-GE-180. Additionally or alternatively, in some embodiments, the imaging technique is SPECT. In some embodiments, localization comprises administering a SPECT reporter to the subject. In some embodiments, the SPECT reporter is 99m Tc-HMPAO. In some embodiments, the therapeutic agent is an anti-tau antibody.

[0163] The present disclosure will be further described in the following examples, which do not limit the scope of the invention described in the claims.

[0164] Example

[0165] Example 1. In vivo rat study to quantify and predict the magnitude of Acoustic and MRI imaging and delivery of therapeutic molecules after blood-brain barrier disruption induced by MRgFUS system Relevance

[0166] The goal of this example is specifically to collect data to inform the correlation between the ΔR2* map and the ΔR1 concentration map and acoustic map when liposomes or iron oxide (or any other alternative) are injected along with DNA or monoclonal antibodies (mAb).

[0167] Male rats (Sprague-Dawley breed) 6 to 8 weeks old and weighing between 150-300 grams were used as test subjects.

[0168] Molecules of various types and sizes are transferred across the BBB after disruption is induced using microbubbles via the ExAblate 4000 system type 2. The sonication pattern is altered to achieve delivery. During such treatments, various real-time feedback / control mechanisms are implemented; real-time cavitation data is used to alter the sonication power profile within the treatment volume as treatment progresses. Various methods are used to assess the passage of molecules across the blood-brain barrier (liposomes or iron oxide (or other alternatives, MRI-based quantification) and DNA (qPCR) or mAb (ELISA) injections).

[0169] Monitor and evaluate one or more of the following:

[0170] Cavitation Acoustic Spectrum. Cavitation is measured using hydrophones, which can be integral to the system transducer or external.

[0171] Abnormalities in T2 and T2* MR images. Multiplanar, multisequence imaging of the brain was performed before and after treatment and during follow-up procedures.

[0172] Gd labeling of liposomal molecules and penetration of Gd across the BBB after treatment were tested. Multiplanar, multisequence T1-weighted imaging of the brain was performed before and after treatment and during follow-up procedures.

[0173] Histopathological assessment of tissue damage (including petechiae, neural injury / effects, immune response, and ischemia) at selected treatment sites.

[0174] Histopathological evaluation of any minor pathological lesions in randomly selected untreated sites.

[0175] This study will recruit a total of 128 animals and has two phases: the first phase includes 20 rats to test the iron oxide dose for the next phase. (Dose range tested: 10-100umol / rat iron oxide), and the second phase includes three groups with two subgroups in each group.

[0176] The first group will test the iron oxide dose found in phase 1, along with DNA injections. Two DNA concentrations will be tested: 0.135 mg / rat and 0.270 mg / rat (18 rats per concentration, 36 rats total).

[0177] The second group will test the iron oxide dose found in phase 1, along with mAb injections. Two mAb concentrations will be tested: 5 mg / kg and 15 mg / kg (18 rats per concentration, 36 rats total).

[0178] The third group will test liposome injections (250 g rats), as well as mAb injections. Two mAb concentrations will be tested: 5 mg / kg and 15 mg / kg (18 rats per concentration, 36 rats total).

[0179] In all three groups, we will treat single or multiple targets with focused ultrasound together with ultrasound contrast agents, possibly with multiple repetitions in the same brain volume. The ultrasound activity of the contrast agent after sonication will be assessed using a hydrophone built into the sonication system or external to it. The intensity and number of further sonications will be decided based on these measurements.

[0180] After the procedure, animals will undergo post-MRI imaging to evaluate treatment success. Specifically, T2-weighted images and T2*-weighted images will be used to evaluate tissue damage, and T2*-weighted imaging and Gd-enhanced T1-weighted imaging will provide information on BBB opening.

[0181] Selected animals may undergo additional examinations. Such examinations may include restoration of the BBB over time and testing for treatment-induced petechiae or edema. Follow-up may be performed up to 4 weeks after treatment. A maximum of 48 hours of shunting is allowed. Both lobes are harvested and, in some cases, sent to pathology without disclosing the parameters of the sonication.

[0182] In selected animals, two lobes were harvested and the selected lobes were sent for pathological examination without disclosing the sonication parameters.

[0183] This study is expected to demonstrate one or more of the following: BBB permeability of liposomal Gd and iron oxide correlates with the permeability of therapeutic molecules (DNA and mAb); T2* signal and acoustic images correlate with the BBB permeability of therapeutic molecules (DNA and mAb); T2 and T2* imaging do not indicate unintentional tissue damage and histopathology does not reveal unintentional microscopic tissue damage.

[0184] Equivalent solutions

[0185] Although the present invention has been described in conjunction with specific embodiments of the present invention, it should be understood that the present invention is capable of further modifications, and this application is intended to cover any variation, use or change that generally follows the principles of the present invention, includes known technologies or commonly used technical means that do not fall within the scope of the present invention but belong to the field to which the present invention belongs, and can be applied to the essential features set forth herein and listed in the scope of the appended claims.

[0186] Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation many equivalents to the specific embodiments specifically described herein. Such equivalents are intended to be encompassed within the scope of the appended claims.

[0187] Incorporated by Reference

[0188] All patents and publications mentioned herein are hereby incorporated by reference in their entirety.

[0189] The publications discussed herein are provided only for their disclosures prior to the filing date of the present application. Anything herein shall not be construed as an admission that the present invention is not entitled to predate such publications by virtue of prior invention. As used herein, all titles are used only for organization and are not intended to limit the present disclosure in any way. The contents of any single section may apply equally to all sections.

Claims

1. A system for destroying a target tissue for treatment, the system comprising: an ultrasonic transducer for sonicating the target volume to cause damage to target tissue therein, thereby increasing its permeability; and A controller, the controller being configured to: (i) monitoring the permeability of the target tissue caused by the destruction; and (ii) at least one of the following: (1) causing the ultrasound transducer to stop sonicating the target volume when the permeability reaches a threshold; (2) presenting the permeability of the target tissue to an operator; and (3) varying ultrasound intensity.

2. The system of claim 1, wherein the permeability corresponds to an upper limit of the size distribution of molecules or molecular complexes that can pass through the damaged target tissue.

3. The system of claim 2, wherein the transparency is calculated based at least in part on: (i) measurement of a molecule that is present in the target tissue under normal conditions but is substantially absent from tissues outside the target tissue, and / or (ii) measurement of the detection marker within the target tissue administered to a subject wherein the permeability is calculated.

4. The system of claim 3, wherein the target tissue is the brain.

5. The system of claim 4, wherein the molecule that is primarily present in the target tissue but substantially absent in tissues outside the target tissue under normal conditions is a brain protein.

6. The system of claim 3, wherein the detection marker and / or the complex is with a polymer, liposome, quantum dot or dextran.

7. The system of claim 3 or claim 6, wherein the detection marker is selected from a magnetic resonance imaging (MRI) visible contrast agent (e.g., ion Gd), a magnetic resonance imaging (MRI) visible contrast agent, a positron emission tomography (PET) reporter (e.g., 18 F-Flubetapyr, 18 F-Fluorobetaben, 18 F-Flutociclib, 18 F-Flumetazol and 18 F-fluorodopa (F-Dopa) and single photon emission computed tomography (SPECT) reporters (e.g. 99m Tc-ECD (ethylcysteine ​​dimer), 123 I-iodoflurane and 99m Tc-TRODAT-1).

8. The system of claim 1, wherein the permeability corresponds to a concentration of molecules or molecular complexes having a certain size range and being able to potentially pass through the target tissue being destroyed.

9. The system of claim 8, wherein the transparency is calculated based on: (i) measurement of molecules within said size range and present in said target tissue but not in tissues outside of said target tissue, and / or (ii) measurement of a detectable marker within said target tissue within said size range and administered to a subject wherein said permeability is calculated.

10. The system of any one of claims 3-9, wherein the permeability is compared between at least two sonication applications administered to the same subject on different days and / or between at least two sonication applications administered to different subjects.

11. The system of claim 10, wherein the permeability is normalized using a measurement of a detection agent or a molecule that is primarily present in the target tissue but substantially absent in tissues outside the target tissue under normal conditions.

12. The system of claim 11, wherein the target tissue is brain, and the molecule that is predominantly present in the target tissue but substantially absent in tissues outside the target tissue under normal conditions is a brain protein.

13. The system of claim 12, wherein the detected NSE or S100B species are homodimers with molecular weights of approximately 78 kDa and 22 kDa, respectively.

14. The system of claim 12, wherein the measurement of the NSE and / or S100B species is used to normalize the delivery of therapeutic agents having a molecular weight range of about 60 kDa to about 100 kDa and / or about 15 kDa to about 30 kDa, respectively.

15. The system of claim 11, wherein the detection marker: conjugated to a polymer within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent, or Complexed with liposomes having a defined size range corresponding to the size range of the therapeutic agent.

16. The system of claim 1, further comprising an MRI device.

17. The system of claim 16, wherein the controller is responsive to the MRI device and is further configured to, based on ΔR1, T1 or The permeability is assessed by measuring at least one of the 18. The system of claim 17, wherein the relationship between the measurement and the permeability is established by calibration.

19. The system of claim 18, wherein the calibration includes a machine learning component.

20. The system of claim 19, wherein the controller is configured to assess the permeability in the absence of a contrast agent.

21. The system of claim 8, wherein the molecule or molecular complex comprises a visualization agent.

22. The system of claim 21, wherein the visualization agent is a contrast agent.

23. The system of claim 22, wherein the contrast agent is ionic Gd (or iron oxide, or phosphate or sulfate derivatives).

24. The system of claim 21, wherein the visualization agent is a fluorophore.

25. The system of claim 8, wherein the size range corresponds to a monoclonal antibody, a viral vector, a liposome, a nucleic acid, or a protein.

26. The system of claim 8, wherein the molecule or molecular complex is a liposome.

27. The system of claim 8, wherein the molecule or molecular complex is a quantum dot.

28. The system of claim 8, wherein the molecule or molecular complex is dextran.

29. The system of claim 8, wherein the molecule or molecular complex is a protein.

30. The system of claim 8, wherein the molecule or molecule complex is a viral vector.

31. The system of claim 8, wherein the molecule or molecular complex is a nucleic acid.

32. The system of claim 1, wherein the transducer is further configured to detect an acoustic signal.

33. The system of claim 32, wherein the controller is responsive to acoustic signals detected by the transducer and is further configured to assess the permeability based on acoustic measurements of target species.

34. The system of claim 33, wherein the target species corresponds to a molecule or a molecular complex coupled to an acoustic agent.

35. The system of claim 33, wherein the controller is further configured to assess the transparency based at least in part on a combination of acoustic measurements and MRI images.

36. The system of claim 33, wherein the controller is further configured to update the target acoustic dose level based at least in part on a combination of acoustic measurements and MRI images.

37. The system of claim 36, wherein the controller is further configured to control ultrasound therapy based at least in part on the updated target acoustic dose level.

38. The system of claim 37, wherein the updated target acoustic dose level is repeatedly used during treatment.

39. The system of claim 34, wherein the permeability corresponds to a concentration of agent-coupled molecules or molecular complexes having a certain size range that have passed through the disrupted target tissue.

40. The system of claim 39, wherein the visualization agent is a contrast agent and the concentration is determined at least in part by a perfusion time or a washout time.

41. The system of claim 40, wherein the cleaning time corresponds to a rise or fall of an acoustic signal characteristic relative to a threshold value.

42. The system of claim 29, wherein the acoustic agent is a contrast agent and the acoustic signal characteristic is a reflection amplitude of an acoustic signal transmitted by the transducer.

43. The system of any one of claims 1-42, further comprising an administration device for introducing microbubbles into the target tissue.

44. A method of destroying a target tissue for treatment, the method comprising the steps of: sonicating the target volume to cause destruction of target tissue therein, thereby increasing its permeability; monitoring the permeability of the target tissue caused by the destruction; and When the permeability reaches a threshold value, sonication of the target volume is stopped.

45. The method of claim 44, wherein the permeability is calculated based at least in part on: (i) measurement of the level of a molecule that is present in the target tissue under normal conditions but is substantially absent from tissues outside the target tissue, in a biological sample from the subject in which the permeability is being monitored, and / or (ii) measurement of a detection marker within the target tissue, wherein the detection marker has been administered to the subject in which the permeability is being monitored.

46. ​​The method of claim 45, wherein the target tissue is brain and / or the biological sample is blood.

47. The method of claim 46, wherein the molecule that is present in the target tissue under normal conditions but is substantially absent from tissue outside the target tissue is a brain protein.

48. The method of claim 47, wherein the NSE or S100B species detected are homodimers with molecular weights of approximately 78 kDa and 22 kDa, respectively.

49. The method of claim 48, wherein the measurement of the NSE and / or S100B species is used to normalize the delivery of therapeutic agents having a molecular weight range of about 60 kDa to about 100 kDa and / or about 15 kDa to about 30 kDa, respectively.

50. The method of claim 45, wherein the detection marker: conjugated to a polymer (e.g., dextran or polyethylene glycol) within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent, or Complexed with liposomes having a defined size range corresponding to the size range of the therapeutic agent.

51. The method of claim 50, wherein the detection marker is selected from the group consisting of a magnetic resonance imaging (MRI) visible contrast agent (e.g., ion Gd), a magnetic resonance imaging (MRI) visible contrast agent, a positron emission tomography (PET) reporter (e.g., 18 F-Flubetapyr, 18 F-Fluorobetaben, 18 F-Flutociclib, 18 F-Flumetazol and 18 F-fluorodopa (F-Dopa) and single photon emission computed tomography (SPECT) reporters (e.g. 99m Tc-ECD (ethylcysteine ​​dimer), 123 I-iodoflurane and 99m Tc-TRODAT-1).

52. The method of claim 44, wherein the sonication comprises at least two applications to the same patient on different days or different applications to different patients.

53. The method of any one of claims 44-52, further comprising introducing microbubbles into the target tissue.

54. A method of comparing the permeability of a target tissue in a subject between at least two focused ultrasound treatment applications on different days, the method comprising: (i) providing a biological sample from said subject, said subject receiving a first application of ultrasound treatment to a target volume to cause damage to said target tissue therein, thereby increasing the permeability thereof, (ii) measuring the amount of a molecule in the biological sample that is present in the target tissue under normal conditions but is substantially absent from tissues outside the target tissue, (iii) monitoring the permeability of the target tissue caused by the destruction; (iv) providing a second biological sample from said subject, said subject having received, on a different date, a second application of ultrasound treatment to the target volume to cause a second disruption of said target tissue therein, thereby increasing a second permeability thereof, (v) measuring the amount of the molecule in the second biological sample that is present in the target tissue under normal conditions but is substantially absent in tissues outside the target tissue, (vi) monitoring the permeability of the target tissue caused by the second disruption; as well as (vii) comparing the permeability of the target tissue in the subject between at least two applications of sonication on different days.

55. The method of claim 54, wherein the target tissue is brain and / or the biological sample is blood.

56. The method of claim 54, wherein the molecule that is present in the target tissue under normal conditions but is substantially absent in tissue outside of the target tissue is a brain protein.

57. The method of claim 56, wherein the NSE or S100B species detected are homodimers with molecular weights of approximately 78 kDa and 22 kDa, respectively.

58. The method of claim 57, wherein the measurement of the NSE and / or S100B species is used to normalize the delivery of therapeutic agents having a molecular weight range of about 60 kDa to about 100 kDa and / or about 15 kDa to about 30 kDa, respectively.

59. The method of any one of claims 54-58, further comprising introducing microbubbles into the target tissue.

60. A method of comparing the permeability of a target tissue in a subject between at least two focused ultrasound therapy applications on different days, the method comprising: (i) providing a subject, said subject receiving a first application of ultrasound treatment to a target volume to cause destruction of said target tissue therein, thereby increasing the permeability thereof, wherein said subject has optionally received a detection marker on the same day, (ii) measuring the level of the detection marker in the target tissue, thereby monitoring the permeability of the target tissue caused by the destruction; (iii) providing a second biological sample from said subject, said subject having received a second application of ultrasound treatment to the target volume on a different day to cause a second disruption of said target tissue therein, thereby increasing a second permeability thereof, optionally wherein said subject has received a detection marker on the same day, (iv) measuring the level of the detection marker in the target tissue, thereby monitoring the permeability of the target tissue caused by the second destruction, thereby monitoring the permeability of the target tissue caused by the second destruction; as well as (v) comparing the permeability of the target tissue in the subject between at least two applications of sonication on different days.

61. The method of claim 60, wherein the target tissue is the brain.

62. The method of claim 60, wherein the detection marker is: conjugated to a polymer (e.g., dextran or polyethylene glycol) within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent, or Complexed with liposomes having a defined size range corresponding to the size range of the therapeutic agent.

63. The method of claim 62, wherein the detection marker is selected from a magnetic resonance imaging (MRI) visible contrast agent, a magnetic resonance imaging (MRI) visible contrast agent, a positron emission tomography (PET) reporter, and a single photon emission computed tomography (SPECT) reporter.

64. The method of claim 63, wherein the MRI visible contrast agent comprises ionic Gd.

65. The method of claim 63, wherein the PET reporter is selected from 18 F-Flubetapyr, 18 F-Fluorobetaben, 18 F-Flutociclib, 18 F-Flumetazol and 18 F-fluorodopa (F-Dopa).

66. The method of claim 63, wherein the SPECT reporter is selected from 99m Tc-ECD (ethylcysteine ​​dimer), 123 I-iodoflurane and 99m Tc-TRODAT-1.

67. The method of any one of claims 60-66, further comprising introducing microbubbles into the target tissue.

68. A method of treating a neurological disease or condition in a subject in need thereof, wherein the neurological disease or condition is characterized by sites in the brain having abnormal production, aggregation and / or deposition of a protein or another biomolecule, wherein a therapeutic agent and / or a microvesicle composition will be, is being or has been administered to the subject, the method comprising the steps of: sonicating a target volume to cause damage to target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and an adjacent blood-brain barrier (BBB); and monitoring the permeability of the target tissue caused by the destruction; and When the permeability reaches a threshold value, sonication of the target volume is stopped, thereby increasing the delivery level of the therapeutic agent to the site compared to a control.

69. The method of claim 68, wherein the control is the level of delivery of the therapeutic agent in the subject who has not received the acoustic pulse sequence and / or the microbubble composition.

70. The method of claim 68, wherein the control is the delivered level of the therapeutic agent in the subject prior to administration of the acoustic pulse sequence and / or the microbubble composition.

71. The method of any one of claims 68-70, wherein the neurological disease or disorder is selected from Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), dementia with Lewy bodies, spinocerebellar ataxia and amyotrophic lateral sclerosis, frontotemporal lobar disease, multiple system atrophy, quadruple repeat tauopathy, and prion disease.

72. The method of any one of claims 68-71, wherein the site is selected from the group consisting of senile plaques, neurofibrillary tangles, neuronal inclusions, Lewy bodies, glial inclusions, cytoplasmic inclusions, and polyglutamine aggregates.

73. The method of any one of claims 68-72, wherein the protein that exhibits abnormal production, aggregation and / or deposition is selected from the group consisting of amyloid-β (Aβ), Tau protein, TDP-43, α-synuclein, FUS / TLS, SOD1 and Huntingtin protein.

74. The method of any one of claims 68-73, wherein the therapeutic agent comprises a small molecule or a biologic drug.

75. The method of claim 74, wherein the therapeutic agent is or comprises a biologic drug.

76. The method of claim 75, wherein the therapeutic agent is selected from a gene therapy agent, an enzyme for enzyme replacement therapy, a vaccine, an antisense oligonucleotide (ASO), a protein therapeutic agent, a modified mRNA agent, and an RNAi agent.

77. The method of claim 75, wherein the therapeutic agent is or comprises an antibody, an antibody-like molecule, or an antigen-binding fragment thereof.

78. The method of claim 77, wherein the therapeutic agent specifically binds to the protein or another biomolecule that exhibits aberrant production, aggregation and / or deposition.

79. The method of claim 77 or claim 78, wherein the therapeutic agent is selected from a nonspecific clearance antibody (e.g., intravenous immunoglobulin, also known as IVIg), an anti-amyloid-β antibody (e.g., aducanumab, gantuzumab, lencanezumab, and donenetumab), an anti-tau antibody (e.g., ceranatumab, gonelizumab, tipranezumab, and zagtenezumab), an anti-TREM2 antibody (e.g., AL002), an anti-α-synuclein antibody (e.g., simpanezumab, putinuzumab, Lu AF82422, ABBV-0805, and MEDI1331), and / or a combination thereof.

80. The method of claim 74, wherein the therapeutic agent is or comprises a small molecule drug.

81. The method of claim 80, wherein the therapeutic agent provides one or more of synaptic plasticity, neuroprotection, reduction of inflammation, neurotransmitter receptor modulation, reduction of oxidative stress.

82. The method of claim 81, wherein the therapeutic agent is selected from donepezil, galantamine, rivastigmine, memantine, sulvorexant, carbidopa-levodopa, selegiline, rasagiline, safinamide, entacapone, benztropine, tolcapone, opicapone, nuprizide, istradefylline, and amantadine, and combinations thereof.

83. The method of any one of claims 68-82, wherein the therapeutic agent is formulated in a liposome.

84. The method of any one of claims 68-83, wherein the therapeutic agent is delivered via a viral vector.

85. The method of any one of claims 68-84, further comprising introducing microbubbles into the target tissue.

86. A method of treating a neurological disease or disorder in a subject in need thereof, wherein the neurological disease or disorder is characterized by sites in the brain having abnormal production, aggregation and / or deposition of a protein or another biomolecule, the method providing a local dose of a therapeutic agent within a predetermined range, the method comprising the steps of: Sonicating a target volume to cause damage to target tissue therein, thereby increasing its permeability, wherein the target volume encompasses the site and an adjacent blood-brain barrier (BBB), wherein the subject has received, will receive, or is receiving, optionally on the same day: a dose of the therapeutic agent, microbubble composition, and / or detection marker; measuring the level of the detection marker in the target tissue, thereby monitoring the local dosage of the therapeutic agent at the site caused by the destruction; When the local dose of the therapeutic agent reaches the predetermined range, sonication of the target volume is stopped.

87. The method of claim 86, wherein the detection marker is: conjugated to a polymer (e.g., dextran or polyethylene glycol) within a defined molecular weight range corresponding to the molecular weight range of the therapeutic agent, or Complexed with liposomes having a defined size range corresponding to the size range of the therapeutic agent.

88. A method as claimed in claim 86 or claim 87, wherein the detection marker is selected from a magnetic resonance imaging (MRI) visible contrast agent, a positron emission tomography (PET) reporter and a single photon emission computed tomography (SPECT) reporter.

89. The method of claim 88, wherein the MRI visible contrast agent comprises ionic Gd.

90. The method of claim 88, wherein the PET reporter is selected from 18 F-Flubetapyr, 18 F-Fluorobetaben, 18 F-Flutociclib, 18 F-Flumetazol and 18 F-fluorodopa (F-Dopa).

91. The method of claim 88, wherein the SPECT reporter is selected from 99m Tc-ECD (ethylcysteine ​​dimer), 123 I-iodoflurane and 99m Tc-TRODAT-1.

92. The method of any one of claims 86-91, wherein the neurological disease or disorder is selected from Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), dementia with Lewy bodies, spinocerebellar ataxia and amyotrophic lateral sclerosis, frontotemporal lobar disease, multiple system atrophy, quadruple repeat tauopathy, and prion disease.

93. The method of any one of claims 86-92, wherein the site is selected from the group consisting of senile plaques, neurofibrillary tangles, neuronal inclusions, Lewy bodies, glial inclusions, cytoplasmic inclusions, and polyglutamine aggregates.

94. The method of any one of claims 86-93, wherein the protein that exhibits abnormal production, aggregation and / or deposition is selected from the group consisting of amyloid-β (Aβ), Tau protein, TDP-43, α-synuclein, FUS / TLS, SOD1 and Huntingtin protein.

95. The method of any one of claims 86-94, wherein the therapeutic agent comprises a small molecule or a biologic drug.

96. The method of claim 95, wherein the therapeutic agent is or comprises a biologic drug.

97. The method of claim 96, wherein the therapeutic agent is selected from a gene therapy agent, an enzyme for enzyme replacement therapy, a vaccine, an antisense oligonucleotide (ASO), a protein therapeutic agent, a modified mRNA agent, and an RNAi agent.

98. The method of claim 96, wherein the therapeutic agent is or comprises an antibody, an antibody-like molecule, or an antigen-binding fragment thereof.

99. The method of claim 98, wherein the therapeutic agent specifically binds to the protein or another biomolecule that exhibits aberrant production, aggregation and / or deposition.

100. The method of claim 98 or claim 99, wherein the therapeutic agent is selected from a nonspecific clearance antibody (e.g., intravenous immunoglobulin, also known as IVIg), an anti-amyloid-β antibody (e.g., aducanumab, gantuzumab, lencanezumab, and donenetumab), an anti-tau antibody (e.g., ceranatumab, gonelizumab, tipranezumab, and zagtenezumab), an anti-TREM2 antibody (e.g., AL002), an anti-α-synuclein antibody (e.g., simpanezumab, putinuzumab, Lu AF82422, ABBV-0805, and MEDI1331), and / or a combination thereof.

101. The method of claim 95, wherein the therapeutic agent is or comprises a small molecule drug.

102. The method of claim 101, wherein the therapeutic agent provides one or more of synaptic plasticity, neuroprotection, reduction of inflammation, neurotransmitter receptor modulation, reduction of oxidative stress.

103. The method of claim 102, wherein the therapeutic agent is selected from donepezil, galantamine, rivastigmine, memantine, suvorexant, carbidopa-levodopa, selegiline, rasagiline, safinamide, entacapone, benztropine, tolcapone, opicapone, nuprizide, istradefylline, and amantadine, and combinations thereof.

104. The method of any one of claims 86-103, wherein the therapeutic agent is formulated in a liposome.

105. The method of any one of claims 86-104, wherein the therapeutic agent is delivered via a viral vector.

106. The method of any one of claims 86-105, further comprising introducing microbubbles into the target tissue.

107. A method of treating a central nervous system infection in a subject in need thereof, wherein a therapeutic agent will be, is being, or has been administered to the subject, the method comprising the steps of: sonicating the target volume to cause destruction of target tissue therein, thereby increasing its permeability; monitoring the permeability of the target tissue caused by the destruction; and When the permeability reaches a threshold value, sonication of the target volume is stopped.

108. The method of claim 107, wherein ceasing the sonication of the target volume increases the delivery level of the therapeutic agent to a site compared to a control.

109. The method of claim 107, wherein stopping the sonication of the target volume comprises stopping the sonication when a local dose of the therapeutic agent reaches a predetermined range.

110. The method of any one of claims 107-109, wherein the therapeutic agent comprises at least one of an antibiotic, an antiviral, an antiretroviral, or an antifungal agent.

111. The method of any one of claims 107-110, further comprising introducing microbubbles into the target volume.

112. A method of treating a congenital enzyme deficiency in a subject in need thereof, wherein a therapeutic agent will be, is being or has been administered to the subject, the method comprising the steps of: sonicating the target volume to cause destruction of target tissue therein, thereby increasing its permeability; monitoring the permeability of the target tissue caused by the destruction; and When the permeability reaches a threshold value, sonication of the target volume is stopped.

113. The method of claim 112, wherein ceasing the sonication of the target volume increases the delivery level of the therapeutic agent to a site compared to a control.

114. The method of claim 112, wherein stopping the sonication of the target volume comprises stopping the sonication when a local dose of the therapeutic agent reaches a predetermined range.

115. The method of any one of claims 112-114, wherein the therapeutic agent comprises enzyme replacement therapy.

116. The method of any one of claims 112-115, further comprising introducing microbubbles into the target volume.

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