Apparatus for measuring microbubble concentration in real time

Abstract

The present disclosure provides a systems and methods for measuring concentration, constituent size, or constituent shape of a composition in real time at the time of a parenteral administration based on the measurement of a physical property of the composition.

Description

APPARATUS FOR MEASURING MICROBUBBLE CONCENTRATION IN REAL TIMEPRIORITY

[0001] This application claims the benefit of, and priority to, U.S. Provisional Application No. 63 / 616,747, filed on December 31, 2024, the contents of which are incorporated by reference in their entirety.FIELD OF DISCLOSURE

[0002] The present disclosure relates, generally, to systems and methods for delivering to patients therapeutics and molecules that are measured in real time.BACKGROUND

[0003] Microbubbles are gas-filled lipid vesicles used as contrast agents in ultrasound applications. During Focused Ultrasound (FUS) treatments, microbubbles are provided to patients through intravenous infusion. The concentration of microbubbles in a patient's blood greatly influences the efficiency and safety of the FUS procedure since microbubbles respond to ultrasound stimulation by producing cavitation.

[0004] Microbubble concentration in the infusion liquid is currently calculated based on the volume of microbubbles added to the total infusion volume using the concentrated microbubble concentration provided by its manufacturer. Microbubble concentration is typically measured by the manufacturer or by the user immediately after microbubble activation, prior to its dilution in the infusion bag. However, the microbubble concentration in the infusion liquid may change during treatment due to microbubble instability, infusion handling, or other factors. Therefore, the determination of the exact dosage of microparticles that is delivered is challenging. Infusion delivery rate in patients was previously suggested to be adjusted based on the acoustic feedback acquired by the FUS system or based on infusion flow rate, but not in response to the actual microbubble concentration present in the infusion liquid.

[0005] Similar issues potentially exist for formulations that are particulate and / or include drugs that have a tendency to aggregate or sediment. Such formulations include liposome, lipid nanoparticles (LNP), some extended release formulations and poly(lactic-co-glycolic acid) (PLGA) particle formulations.

[0006] Therefore, new methods of determining drug concentration in real time at the time of infusion are needed.SUMMARY

[0007] Accordingly, the present disclosure relates to, inter alia, systems and methods for determining concentration of composition in real time at the time of infusion and for providing an updated rate of administration of drugs based, inter alia, on the determined concentration of composition.

[0008] In aspects, the present disclosure provides a system for measuring concentration of a composition in real time at the time of a parenteral administration. In some embodiments, the system comprises (a) an apparatus capable of measuring a physical property of the composition, the apparatus comprising a detector; (b) a controller, operably coupled to the detector, and (c) an outlet being connected to a port of administration of the composition to a subject. In some embodiments, the controller is configured to store a desired concentration of the composition selected by an operator, optionally a human operator. In some implementations, the system further comprises an inlet from a reservoir of the pharmaceutical composition into an apparatus. In some embodiments, the controller is configured to measure the physical property of the composition. In some embodiments, the controller is configured to adjust a flow rate of the composition in an outlet. In some embodiments, the controller adjusts the flow rate compared to an initial flow rate selected by the operator. In some embodiments, the controller adjusts the flow rate compared to the flow rate at which the composition is being administered. In some embodiments, the controller adjusts the flow rate based on a standard curve providing a correlation between test measurements and test concentrations. In some embodiments, the controller adjusts the flow rate based on an equation providing a correlation between test measurements and test concentrations. In some embodiments, the controller is configured to adjust a flow rate of the composition in the outlet, based on an algorithm that analyzes the condition of the treated organ / body region and adjust the concentration of the composition accordingly. In embodiments, the condition of the treated organ / body region is provided to the controller by an operator, optionally a human operator. In some embodiments, the condition of the treated organ / body region provided to the controller by an additional piece of medical equipment connected to the controller. In some embodiments, the condition of the treated organ\body region provided to the controller by an additional piece ofmedical equipment connected to the controller. In some embodiments, the physical property is selected from light absorbance, fluorescence and phosphorescence. In some embodiments, the apparatus comprises a light source and a detector of light. In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0009] In aspects, the present disclosure provides a system for measuring concentration of a composition in real time at the time of a parenteral administration. In some embodiments, the system comprises (a) an apparatus capable of measuring a physical property of the composition selected from light absorbance, fluorescence and phosphorescence, the apparatus comprising a light source and a detector of light; (b) a controller, operably coupled to the detector; and (c) an outlet being connected to a port of administration of the composition to a subject. In some embodiments, the controller is configured to store a desired concentration of the composition selected by an operator, optionally a human operator. In some embodiments, the controller is configured to measure the physical property of the composition. In some embodiments, the controller is configured to adjust a flow rate of the composition in an outlet. In some embodiments, the controller adjusts the flow rate based on a standard curve providing a correlation between test measurements and test concentrations. In some embodiments, the controller adjusts the flow rate based on an equation providing a correlation between test measurements and test concentrations. In some embodiments, the controller is configured to adjust a flow rate of the composition in an outlet. In some embodiments, the controller adjusts the flow rate compared to an initial flow rate selected by the operator. In some embodiments, the controller adjusts the flow rate compared to the flow rate at which the composition is being administered. In some embodiments, the controller is configured to adjust a flow rate of the composition in the outlet, based on an algorithm that analyzes the condition of the treated organ\body region and adjust the concentration of the composition accordingly. In embodiments, the condition of the treated organ / body region is provided to the controller by an operator, optionally a human operator. In some embodiments, the condition of the treated organ / body region provided to the controller by an additional piece of medical equipment connected to the controller.

[0010] In some embodiments, the light source is or comprises a light emitting diode (LED). In some embodiments, the detector of light is or comprises a light emitting diode (LED). In someembodiments, the light is infrared (IR) light. In some embodiments, the light source is an IR LED. In some embodiments, the detector of light is an IR LED.

[0011] In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the composition is or comprises a pharmaceutical composition. In some embodiments, the reservoir of the composition is an infusion bag. In some embodiments, the reservoir of the composition is a drip chamber.

[0012] In some embodiments, the parenteral administration is infusion.

[0013] In some embodiments, the detector provides an output of concentration of the composition. In some embodiments, the output is or comprises a print out and / or an on-screen display.

[0014] In some embodiments, the apparatus and / or the controller comprise a manual control unit configured to manually adjust the flow rate. In some embodiments, the flow rate is manually adjusted by a human operator. In some embodiments, the flow rate is manually adjusted by a human operator based on the output of concentration of the composition.

[0015] In some embodiments, the apparatus and / or the controller comprise an automated control unit configured to automatically adjust the flow rate. In some embodiments, the apparatus and / or the controller automatically adjust the flow rate based on the output of concentration of the composition and / or human input. In some embodiments, the apparatus and / or the controller automatically increase the flow rate compared to initial flow rate when the concentration of the composition is lesser compared to the desired concentration. In some embodiments, the apparatus and / or the controller automatically decrease the flow rate compared to initial flow rate when the concentration of the composition is greater compared to the desired concentration. In some embodiments, the apparatus and / or the controller automatically maintain the flow rate substantially equal to initial flow rate when the concentration of the composition is substantially equal compared to the desired concentration. In some embodiments, the apparatus or the controller automatically adjust the flow rate of the composition, based on an algorithm that analyzes the condition of the treated organ / body area and adjust the pharmaceutical concentration accordingly.

[0016] In some embodiments, the composition comprises a sedimentable ingredient. In some embodiments, the sedimentable ingredient is selected from a microbubble, a lipid nanoparticle (LNP) or a liposome.

[0017] In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the microbubble composition comprises one or more lipid-based microspheres. In some embodiments, the microbubble composition is perflutren lipid microspheres. In some embodiments, the microbubble composition comprises (R)-hexadecanoic acid, l-[(phosphonoxy)methyl]-l,2-ethanediyl ester, monosodium salt (DPPA); (R)-4-hydroxy- N,N,Ntrimethyl-10-oxo-7-[(l-oxohexadecyl)oxy]-3,4,9-trioxa-4-phosphapentacosan-l-aminium, 4-oxide, inner salt (DPPC); (R)-a-[6-hydroxy-6-oxido-9-[(l-oxohexadecyl)oxy]-5,7,l l-trioxa-2- aza-6-phosphahexacos-l-yl]-co-methoxypoly(ox-l,2-ethanediyl), monosodium salt; and (N- (methoxypolyethylene glycol 5000 carbamoyl)-l,2-dipalmitoyl-sn-glycero-3- phosphatidylethanolamine, monosodium salt, MPEG5000 DPPE). In some embodiments, the microbubble compositions are lipid-coated echogenic microbubbles filled with octafluoropropane gas. In some embodiments, the microbubble compositions comprise octafluoropropane encapsulated in an outer lipid shell comprising (R)-hexadecanoic acid, l-[(phosphonoxy)methyl]- 1,2-ethanediyl ester, monosodium salt (DPPA); (R)-4-hydroxy-N,N,Ntrimethyl-10-oxo-7-[(l- oxohexadecyl)oxy]-3,4,9-trioxa-4-phosphapentacosan-l-aminium, 4-oxide, inner salt (DPPC); (R)-a-[6-hydroxy-6-oxido-9-[(l-oxohexadecyl)oxy]-5,7,l l-trioxa-2-aza-6-phosphahexacos-l- yl]-co-methoxypoly(ox-l,2-ethanediyl), monosodium salt; and (N-(methoxypoly ethylene glycol 5000 carbamoyl)- l,2-dipalmitoyl-sn-glycero-3- phosphatidylethanolamine, monosodium salt, MPEG5000 DPPE).

[0018] In some embodiments, the composition comprises or further comprises a biologic drug selected from an antibody, a virus, a vaccine composition, a protein therapeutic, and a nucleic acid therapeutic selected from RNA, mRNA, chemically modified mRNA, a small interfering RNA (siRNA), a microRNA (miRNA), and an antisense RNA. In some embodiments, the composition comprises or further comprises a small molecule drug.

[0019] In some embodiments, the apparatus and / or the controller communicate with one or more additional piece of medical equipment. In some embodiments, the apparatus and / or the controller adjusts the desired concentration based on an input from the additional piece of medical equipment.In some embodiments, the apparatus and / or the controller provide an input to the additional piece of medical equipment to control its function.

[0020] In some embodiments, the additional piece of medical equipment is an ultrasound transducer. In some embodiments, the input from the apparatus and / or the controller controls the kind of an ultrasound beam, a shape of an ultrasound beam, a pulse pattern of an ultrasound beam, a pulse duration of an ultrasound beam, a pulse repetition frequency (PRF) of an ultrasound beam, the center frequency of ultrasound beam, a spatial peak temporal average acoustic intensity (Ispta), a spatial peak pulse average acoustic intensity (Isppa), a power of the ultrasound beam, and / or the duration of sonication.

[0021] In aspects, the present disclosure provides a method for measuring concentration of a composition in real time at the time of a parenteral administration, the method comprising: (a) providing a composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, (c) connecting the system to a port of administration of the composition to the subject, (d) measuring the physical property by the detector; and (e) determining the concentration of the composition. In some embodiments, the composition is or comprises a microbubble compositions. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0022] In some embodiments, the determining is performed using a standard curve of test measurements of the physical property and test concentrations of the composition. In some embodiments, the standard curve is saved in the controller.

[0023] In some embodiments, the determining is performed using an equation. In some embodiments, the equation provides a correlation between test measurements and test concentrations to determine the concentration. In some embodiments, the equation is saved in the controller.

[0024] In some embodiments, the physical property is selected from light absorbance, fluorescence and phosphorescence.

[0025] In aspects, the present disclosure provides a method of administering a composition to a subject. In some embodiments, the method comprises (a) providing a composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, and (c)connecting the system to a port of administration of the composition to the subject. In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0026] In aspects, the present disclosure provides a method of treating a subject in need thereof suffering from a disease or disorder. In some embodiments, the method comprises a step of administering a composition to the subject. In some embodiments, the step of administering comprises (a) providing the composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, and (c) connecting the system to a port of administration of the composition to the subject. In some embodiments, the method further comprises administering an ultrasound beam to the subject and optionally a second composition that is capable of preventing or treating the disease or disorder. In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0027] In aspects, the present disclosure provides a method of treating a subject in need thereof suffering from a disease or disorder that is capable of being treated with ultrasound. In some embodiments, the method comprises a step of administering a composition. In some embodiments, the step of administering comprises (a) providing a composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, and (c) connecting the system to a port of administration of the composition to the subject. In some embodiments, the method further comprises administering an ultrasound beam to the subject. In some embodiments, the method further comprises administering an ultrasound beam to the subject and a second composition that is capable of preventing or treating the disease or disorder. In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0028] In some embodiments, the composition comprising microbubbles is administered immediately before and / or during the application of the ultrasound beam. In some embodiments, the composition comprising microbubbles is administered contemporaneously with the application of the ultrasound beam.

[0029] In some embodiments, the disease or disorder is selected from a neurological condition. In some embodiments, the neurological condition is selected from Alzheimer’s Disease,Parkinson's Disease, essential tremor, neuropathic pain, epilepsy, addiction, post-traumatic stress disorder, depression and obsessive-compulsive disorder. In some embodiments, the disease or disorder is selected from a musculoskeletal pathology. In some embodiments, the musculoskeletal pathology is selected from osteoarthritis, soft tissue shoulder pathology, and myofascial pain. In some embodiments, the disease or disorder is selected from a cancer. In some embodiments, the cancer is a brain tumor or a metastatic brain tumor. In some embodiments, the disease or disorder is selected from a cardiovascular condition. In some embodiments, the cardiovascular condition is selected from varicose veins, blood clots, calcified arteries, and calcified veins. In some embodiments, the disease or disorder is selected from a kidney condition. In some embodiments, the kidney condition is a kidney stone. In some embodiments, the disease or disorder is selected from an eye condition. In some embodiments, the eye condition is selected from dry eye disease, glaucoma, presbyopia, macular degeneration, retinal impairment, and retinal injury. In some embodiments, wherein the second composition to the subject, wherein the second composition comprises a pharmaceutical agent that is used for treating the neurological condition, the musculoskeletal condition, the cancer, the cardiovascular condition, the kidney condition, or the eye condition.BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In the drawings, like reference characters generally refer to the same parts throughout the different views. Also, the drawings are not necessarily to scale, with an emphasis instead generally being placed upon illustrating the principles of the disclosure. In the following description, various embodiments of the present disclosure are described with reference to the following drawings.

[0031] FIG. 1A schematically depicts a non-limiting and exemplary infusion drip controller system in accordance with various embodiments of the current disclosure.

[0032] FIG. IB schematically depicts a non-limiting and exemplary infusion drip controller system containing a microbubble concentration measurement apparatus in accordance with various embodiments of the current disclosure.

[0033] FIG. 2A and FIG. 2B describe various non-limiting embodiments describing the flow process in which this disclosure can be implemented. FIG. 2A is a non-limiting flow chart illustrating various options after the microbubble concentration measurer registers actualmicrobubble concentration. FIG. 2B is a non-limiting flow chart illustrating various options after the microbubble concentration measurer registers microbubble concentrations that are as desired, higher than decided or lower than desired.

[0034] FIG. 3 is a line graph of the generated voltages as detected by a non-limiting infrared (IR) device of the present disclosure as a function of microbubble (MB) concentrations measured using a Counter coulter and.

[0035] FIG. 4 is a line graph showing of the stability of microbubble formulation, as determined using the generated voltage as a function of time after microbubble activation.

[0036] FIG. 5A and FIG. 5B show the changes in MB diameter as measured using IR measurements (FIG. 5 A) and a Coulter Counter (FIG. 5B).DETAILED DESCRIPTION

[0037] The present disclosure based, in part, on the discovery of a system and apparatus for measuring drug concentration in real time at the time of administering the drug to a patient, e.g., by infusion.

[0038] Microbubbles used in Focused Ultrasound (FUS) treatments are provided to patients through intravenous infusion. The concentration of microbubbles in a patient’s blood greatly influences the efficiency and safety of the FUS procedure since microbubbles respond to ultrasound stimulation by producing cavitation. Microbubble concentration in the infusion liquid is currently calculated based on the volume of microbubbles added to the total infusion volume using the concentrated microbubble concentration provided by its manufacturer. However, as shown in FIG. 4, the microbubble concentration in the infusion liquid may change during treatment due to microbubble instability, infusion handling, or other factors.

[0039] Today, microbubble concentration is measured by their manufacturer or by the user immediately after microbubble activation. The final microbubble concentration in the infusion bag is not typically measured. The infusion delivery rate in patients was previously suggested to be adjusted based on the acoustic feedback acquired by the FUS system or based on infusion flow rate, but not in response to the actual microbubble concentration present in the infusion liquid. This is because there hasn’t been a sufficient reason provided for measuring the microbubble concentration in real time at the time of infusion, as provided by FIG. 4.

[0040] Currently, there are four main ways in which microbubble concentration is measured. The methods that are currently available include the use of Coulter counter, which measures temporal changes in the impedance of a liquid, which is proportional to a particle size. Another method for measuring microbubble concentration relies on dynamic light scattering (DLS), which measures light scattering, which is related to a particle size. Yet another method for measuring microbubble concentration depends on microscope / camera for visual counting of the number and size of microbubbles in a specific volume. Nanoparticle Tracking Analysis (NTA), which utilizes the properties of both light scattering and Brownian motion in order to obtain the nanoparticle size distribution of samples in liquid suspension, is also for used for measuring microbubble concentration. During NTA, particles in liquid suspension are loaded into a sample chamber, which is illuminated by a specially shaped laser beam. See Sennoga et al., Evaluation of methods for sizing and counting of ultrasound contrast agents Ultrasound in Medicine & Biology, , 38(5): 834-845 (2012); Koo et al., Characterizing how size distribution and concentration affect echogenicity of ultrasound contrast agents, Ultrasonics, 127: 106827(2023); H. Mulvana et al., Characterization of Contrast Agent Microbubbles for Ultrasound Imaging and Therapy Research. IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control, 64(1): 232-251 (2017); Luypaert et al., Near-infrared spectroscopy applications in pharmaceutical analysis. Taianta 72(3), 865-883 (2007); and Lakshmanan et al., Preparation of biogenic gas vesicle nanostructures for use as contrast agents for ultrasound and MRI, Nature protocols 1 (10), 2050- 2080 (2017), the entire contents of which are incorporated by reference. While these methods might offer better precision and can also measure microbubble size distribution, they are expensive and require big machines, which are not appropriate to measure microbubble concentration in the infusion liquid in real time. The visual / Microscopy technique is cheaper, but it is time consuming. It is noted that none of these methods provide for measurement of the microbubble concentration in real time at the time of infusion.System for Measuring Concentration of Composition

[0041] In aspects, the present disclosure provides a system for measuring concentration of a composition in real time at the time of a parenteral administration. In some embodiments, the system comprises (a) an apparatus capable of measuring a physical property of the composition, the apparatus comprising a detector; (b) a controller, operably coupled to the detector, and (c) an outlet being connected to a port of administration of the composition to a subject. In someimplementations, the system further comprises an inlet from a reservoir of the pharmaceutical composition into an apparatus. In some embodiments, the controller is configured to store a desired concentration of the composition selected by an operator, optionally a human operator. In some embodiments, the controller is configured to measure the physical property of the composition. In some embodiments, the controller is configured to adjust a flow rate of the composition in an outlet. In some embodiments, the controller adjusts the flow rate compared to an initial flow rate selected by the operator. In some embodiments, the controller adjusts the flow rate compared to the flow rate at which the composition is being administered. In some embodiments, the controller adjusts the flow rate based on a standard curve providing a correlation between test measurements and test concentrations. In some embodiments, the controller uses a standard curve of test measurements of the physical property and test concentrations of the composition to determine the concentration. In some embodiments, the standards curves are predetermined and saved in the controller (FIG. 3). In some embodiments, the controller adjusts the flow rate based on an equation providing a correlation between test measurements and test concentrations. In some embodiments, the equations having one or more variables are saved in the detector. Illustrative non-limiting variables include the measurements of the physical property of a buffer only controls, and the measurements of the physical property of compositions of pre-defined concentrations. In some embodiments, the controller uses a standard curve of test measurements of the physical property and test concentrations of the composition to determine the concentration. In some embodiments, the standards curves are predetermined and saved in the controller (FIG. 3). In some embodiments, the controller is configured to adjust a flow rate of the composition in the outlet compared to an initial flow rate selected by the operator. In some embodiments, the controller is configured to adjust a flow rate of the composition in the outlet, based on an algorithm that analyzes the condition of the treated organ / body region and adjust the concentration of the composition accordingly. In embodiments, the condition of the treated organ / body region is provided to the controller by an operator, optionally a human operator. In some embodiments, the condition of the treated organ / body region provided to the controller by an additional piece of medical equipment connected to the controller. In some embodiments, the additional piece of medical equipment is selected from an ultrasound transducer device a computer tomography (CT) device, a magnetic resonance imaging (MRI) device, a positron emission tomography (PET) device, a single-photon emission computed tomography (SPECT) device, an ultrasonography device, a functional MRI(fMRI) device, an X-ray fluoroscopy device, a diffusion-tensor imaging (DU) device, an angiography device (including a magnetic resonance angiography (MRA) device), a magnetoencephalography (MEG) device, an electroencephalography (EEG) device, a nearinfrared spectroscopy (NIRS) device, an optical imaging device, a bone densitometry device and a combination of two or more thereof.

[0042] In some embodiments, the physical property is selected from light absorbance, fluorescence and phosphorescence. In some embodiments, the apparatus comprises a light source and a detector of light. In some embodiments, the composition is or comprises a microbubble compositions. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0043] In aspects, the present disclosure provides a system for measuring concentration of a composition in real time at the time of a parenteral administration. In some embodiments, the system comprises (a) an apparatus capable of measuring a physical property of the composition selected from light absorbance, fluorescence and phosphorescence, the apparatus comprising a light source and a detector of light; (b) a controller, operably coupled to the detector; and (c) an outlet being connected to a port of administration of the composition to a subject. In some implementations, the system further comprises an inlet from a reservoir of the pharmaceutical composition into an apparatus. In some embodiments, the controller is configured to store a desired concentration of the composition selected by an operator, optionally a human operator. In some embodiments, the controller is configured to measure the physical property of the composition. In some embodiments, the controller is configured to adjust a flow rate of the composition in an outlet. In some embodiments, the controller adjusts the flow rate compared to an initial flow rate selected by the operator. In some embodiments, the controller adjusts the flow rate compared to the flow rate at which the composition is being administered. In some embodiments, the controller adjusts the flow rate based on a standard curve providing a correlation between test measurements and test concentrations. In some embodiments, the standards curves are predetermined and saved in the controller. In some embodiments, the controller uses an equation providing a correlation between test measurements and test concentrations to determine the concentration. In some embodiments, the controller adjusts the flow rate based on an equation providing a correlation between test measurements and test concentrations. In some embodiments, the equations having one or more variables are saved in the detector. Illustrative non-limiting variables includeabsorbance, fluorescence and / or phosphorescence of a buffer only controls, and absorbance, fluorescence and / or phosphorescence of compositions of pre-defined concentrations. In some embodiments, the standards curves are predetermined and saved in the detector. In some embodiments, the controller is configured to (iii) adjust a flow rate of the composition in an outlet. In some embodiments, the controller adjusts the flow rate compared to an initial flow rate selected by the operator. In some embodiments, the controller adjusts the flow rate compared to the flow rate at which the composition is being administered. In some embodiments, the controller is configured to adjust a flow rate of the composition in the outlet, based on an algorithm that analyzes the condition of the treated organ / body region and adjust the concentration of the composition accordingly. In embodiments, the condition of the treated organ / body region is provided to the controller by an operator, optionally a human operator. In some embodiments, the condition of the treated organ / body region provided to the controller by an additional piece of medical equipment connected to the controller. In some embodiments, the additional piece of medical equipment is selected from an ultrasound transducer device a computer tomography (CT) device, a magnetic resonance imaging (MRI) device, a positron emission tomography (PET) device, a single-photon emission computed tomography (SPECT) device, an ultrasonography device, a functional MRI (fMRI) device, an X-ray fluoroscopy device, a diffusion-tensor imaging (DTI) device, an angiography device (including a magnetic resonance angiography (MRA) device), a magnetoencephalography (MEG) device, an electroencephalography (EEG) device, a near-infrared spectroscopy (NIRS) device, an optical imaging device, a bone densitometry device and a combination of two or more thereof.

[0044] In some embodiments, the light source is or comprises a light emitting diode (LED). In some embodiments, the detector of light is or comprises a light emitting diode (LED). In some embodiments, the light is infrared (IR) light. In some embodiments, the light source is an IR LED. In some embodiments, the detector of light is an IR LED.

[0045] In some embodiments, the reservoir of the composition is an infusion bag. In some embodiments, the reservoir of the composition is a drip chamber.

[0046] In some embodiments, the parenteral administration is infusion.

[0047] In some embodiments, the detector provides an output of concentration of the composition. In some embodiments, the output is or comprises a print out and / or an on-screen display.

[0048] In some embodiments, the apparatus and / or the controller comprise a manual control unit configured to manually adjust the flow rate. In some embodiments, the flow rate is manually adjusted by a human operator. In some embodiments, the flow rate is manually adjusted by a human operator based on the output of concentration of the composition (FIG. 2A and FIG. 2B).

[0049] In some embodiments, the apparatus and / or the controller comprise an automated control unit configured to automatically adjust the flow rate (FIG. 2A and FIG. 2B). In some embodiments, the apparatus and / or the controller automatically adjust the flow rate based on the output of concentration of the composition and / or human input. In some embodiments, the apparatus and / or the controller automatically increase the flow rate compared to initial flow rate when the concentration of the composition is lesser compared to the desired concentration. In some embodiments, the apparatus and / or the controller automatically decrease the flow rate compared to initial flow rate when the concentration of the composition is greater compared to the desired concentration. In some embodiments, the apparatus and / or the controller automatically maintain the flow rate substantially equal to initial flow rate when the concentration of the composition is substantially equal compared to the desired concentration (FIG. 2A and FIG. 2B).

[0050] In some embodiments, the composition comprises a sedimentable ingredient. In some embodiments, the sedimentable ingredient is selected from a microbubble, a lipid nanoparticle (LNP) or a liposome.

[0051] In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the microbubble composition comprises one or more lipid-based microspheres. In some embodiments, the microbubble composition is perflutren lipid microspheres. In some embodiments, the microbubble composition comprises (R)-hexadecanoic acid, l-[(phosphonoxy)methyl]-l,2-ethanediyl ester, monosodium salt (DPPA); (R)-4-hydroxy- N,N,Ntrimethyl-10-oxo-7-[(l-oxohexadecyl)oxy]-3,4,9-trioxa-4-phosphapentacosan-l-aminium, 4-oxide, inner salt (DPPC); (R)-a-[6-hydroxy-6-oxido-9-[(l-oxohexadecyl)oxy]-5,7,l l-trioxa-2- aza-6-phosphahexacos-l-yl]-co-methoxypoly(ox-l,2-ethanediyl), monosodium salt; and (N- (methoxypolyethylene glycol 5000 carbamoyl)-!, 2-dipalmitoyl-sn-glycero-3-phosphatidylethanolamine, monosodium salt, MPEG5000 DPPE). In some embodiments, the microbubble compositions are lipid-coated echogenic microbubbles filled with octafluoropropane gas. In some embodiments, the microbubble compositions comprise octafluoropropane encapsulated in an outer lipid shell comprising (R)-hexadecanoic acid, l-[(phosphonoxy)methyl]- 1,2-ethanediyl ester, monosodium salt (DPPA); (R)-4-hydroxy-N,N,Ntrimethyl-10-oxo-7-[(l- oxohexadecyl)oxy]-3,4,9-trioxa-4-phosphapentacosan-l-aminium, 4-oxide, inner salt (DPPC); (R)-a-[6-hydroxy-6-oxido-9-[(l-oxohexadecyl)oxy]-5,7,l l-trioxa-2-aza-6-phosphahexacos-l- yl]-co-methoxypoly(ox-l,2-ethanediyl), monosodium salt; and (N-(methoxypoly ethylene glycol 5000 carbamoyl)- l,2-dipalmitoyl-sn-glycero-3- phosphatidylethanolamine, monosodium salt, MPEG5000 DPPE).

[0052] In some embodiments, the microbubbles comprise a shell and a core. In some embodiments, the microbubbles core may be a gas core. In some embodiments, the gas core comprises one or a combination of nitrogen, oxygen, carbon dioxide, hydrogen, inert gas, helium, argon, xenon, krypton, a sulphur fluoride, selenium hexafluoride, a halogenated silane, a hydrocarbon containing up to 7 carbon atoms, an alkane, methane, ethane, a propane, a butane, a pentane, a cycloalkane, cyclopropane, cyclobutene, cyclopentane, an alkene, ethylene, propene, propadiene, a butene, an alkyne, acetylene, propyne, an ether, dimethyl ether, a ketone, an ester, or a halogenated hydrocarbon containing up to 7 carbon atoms. In some embodiments the halogenated silane comprises methylsilane or dimethylsilane. In some embodiments the sulphur fluoride comprises sulphur hexafluoride, disulphur decafluoride, trifluoromethylsulphur pentafluoride.

[0053] In some embodiments, the microbubble gas core comprises a perfluorocarbon or other perflourinated gas. In some embodiments, the perflourinated gas comprises any one of: bromochlorodifluoromethane, chlorodifluoromethane, dichlorodifluoro-methane, bromotrifluoromethane, chlorotrifluoromethane, chloropenta- fluoroethane, dichlorotetrafluoroethane, chlorotrifluoroethylene, fluoroethylene, sulphur hexafluoride, ethylfluoride, or 1,1- difluoroethane. In some embodiments the perfluorocarbon comprises a perfluoroalkane, a perfluoropropane, a perfluorobutane, a perfluoropentane, a perfluorohexane, a perfluoroheptanes, a perfluoroalkene, a perfluoroalkynes, or a perfluorocycloalkane. In some embodiments the perfluoroalkane comprises perfluoromethane or perfluoroethane. In some embodiments the perfluorobutane comprises perfluoro-n-butane. In some embodiments theperfluoro-n-butane comprises an admixture with other isomers. In some embodiments the other isomers comprise perfluoro-iso-butane.

[0054] In some embodiments, the microbubble shell comprises a polymer. In some embodiments, the polymer comprises any one of polyvinyl alcohol (PVA), polyethylene glycol (PEG), poly(iso- butylcyanoacrylate), Poly(lactic-co-glycolic acid) (PLGA), poly(allylamine hydrochloride) (PAH) and poly (styrene sulfonate) (PSS).

[0055] In some embodiments, the microbubble shell comprises a lipid. In some embodiments, the lipid comprises a phospholipid. In some embodiments the phospholipid comprises any one of 1,2- dimyristoyl-sn-glycerol-3-phosphocholine (DMPC), 4-dimethylaminochalcone (DMAC), dipalmitoyl phosphatidylcholine (DPPC), l,2-distearyol-sn-glycero-3-phosphocoline (DSPC), and l,2-distearyol-sn-glycero-3-phosphoethanolamine-N-[methoxy(poly ethylene glycol)2000] (DSPE-PEG2000).

[0056] In some embodiments, the composition is or comprises a pharmaceutical composition. In some embodiments, the composition is or comprises one or more microbubble compositions. In some embodiments, one or more microbubble compositions are administered immediately before and / or during the application of the ultrasound beam.

[0057] In some embodiments, one or more microbubble compositions are administered contemporaneously with the application of the ultrasound beam.

[0058] In some embodiments, microbubble compositions comprise one or more lipid-based microspheres. In some embodiments, microbubble compositions are perflutren lipid microspheres. In some embodiments, microbubble compositions comprise (R)-hexadecanoic acid, 1- [(phosphonoxy)methyl]-l,2-ethanediyl ester, monosodium salt (DPP A); (R)-4-hydroxy- N,N,Ntrimethyl-10-oxo-7-[(l-oxohexadecyl)oxy]-3,4,9-trioxa-4-phosphapentacosan-l-aminium, 4-oxide, inner salt (DPPC); (R)-a-[6-hydroxy-6-oxido-9-[(l-oxohexadecyl)oxy]-5,7,l l-trioxa-2- aza-6-phosphahexacos-l-yl]-co-methoxypoly(ox-l,2-ethanediyl), monosodium salt; and (N- (methoxypolyethylene glycol 5000 carbamoyl)-l,2-dipalmitoyl-sn-glycero-3- phosphatidylethanolamine, monosodium salt, MPEG5000 DPPE). In some embodiments, microbubble compositions are lipid-coated echogenic microbubbles filled with octafluoropropane gas. In some embodiments, microbubble compositions comprise octafluoropropane encapsulated in an outer lipid shell comprising (R)-hexadecanoic acid, l-[(phosphonoxy)methyl]-l,2-ethanediylester, monosodium salt (DPPA); (R)-4-hydroxy-N,N,Ntrimethyl-10-oxo-7-[(l- oxohexadecyl)oxy]-3,4,9-trioxa-4-phosphapentacosan-l-aminium, 4-oxide, inner salt (DPPC); and (R)-a-[6-hydroxy-6-oxido-9-[(l-oxohexadecyl)oxy]-5,7,l l-trioxa-2-aza-6-phosphahexacos- l-yl]-co-methoxypoly(ox-l,2-ethanediyl), monosodium salt; (N-(methoxypolyethylene glycol 5000 carbamoyl)- l,2-dipalmitoyl-sn-glycero-3 -phosphatidylethanolamine, monosodium salt, MPEG5000 DPPE).

[0059] In some embodiments, the microbubbles comprise a phospholipid-based ultrasound contrast agent comprising one or more of dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine-PEG(5,000), and dipalmitoylphosphatidic acid. Illustrative microbubbles that are suitable in the present methods include but are not limited to commercially available preparations such as DEFINITY®, SONOVUE®, and OPTISON™.

[0060] In some embodiments, microbubble compositions are administered to the patient no more than 60, or 30, or 20, or 10 minutes before the application of the ultrasound beam.

[0061] In some embodiments, the microbubble compositions are administered to the patient throughout the method.

[0062] In some embodiments, microbubble compositions are administered by systemic injection, bolus injection or slow diffusion injection. In some embodiments, the microbubble compositions are delivered into systemic circulation as a continuous intravenous infusion. In some embodiments, the continuous intravenous infusion of the microbubble compositions allows for a steady state microbubble concentration in the blood stream. In additional or alternative embodiments, the microbubble compositions are administration of one or more acute bolus injections. In some embodiments, the acute bolus injections of the microbubble compositions provide for a transient and high concentrations of the microbubbles in the bloodstream that may be timed relative to the administration of an ultrasound beam. In additional or alternative embodiments, the microbubble compositions are co-formulated with one or more therapeutic agents. In these embodiments, the administration of the ultrasound beam opens the blood-brain barrier in combination of microbubbles and the co-formulated therapeutic agent is preferentially released at the site of the opened blood-brain barrier. In some embodiments, microbubble compositions are administered by systemic infusion. In some embodiments, microbubble compositions are administered by continuous intravenous infusion. In embodiments, microbubblecompositions are administered by continuous intravenous infusion of a mixture about 1 x IO10to about 6 x IO10microsphere in a carrier. In embodiments, microbubble compositions are administered by continuous intravenous infusion of a mixture about 1 x 1010to about 6 x 1010microsphere in a saline carrier. In embodiments, microbubble compositions are administered by continuous intravenous infusion of a mixture about 1 x IO10to about 2 x 1011microsphere in about 250 mL of saline carrier. In embodiments, microbubble compositions are administered by continuous intravenous infusion of about 4 x 107to about 8 x 108 / mL. In embodiments, microbubble compositions are continuously infused at a rate of about 1 to about 10 mL / minute during the application of the ultrasound beam.

[0063] In some embodiments, the present disclosure relates, in part, to the present ultrasound applications in conjunction with microbubble alternatives, such as phase-shift droplets, nanobubbles, nanodroplets or nanoparticles, e.g., gold nanoparticles. Embodiments relating to microbubbles apply equally to nanodroplets or nanoparticles, e.g., gold nanoparticles.

[0064] In some embodiments, the microbubble compositions are administered to the patient throughout the method. In some embodiments, the microbubble compositions are administered by systemic injection, bolus injection or slow diffusion injection. In some embodiments, the microbubble compositions are administered by systemic infusion. In some embodiments, substantially all of the disrupted BBB closes after the application of the ultrasound beam. In some embodiments, the transient disruption of the BBB allows for movement of the antibody-based treatment agent across the BBB. In some embodiments, the human patient demonstrates an increased standard uptake value ratio (SUVr) of greater than about 50%, or greater than about 60%, or greater than about 70%, or greater than about 80%, or greater than about 90%, as compared to administration of the antibody-based treatment agent in the absence of application of the focused ultrasound beam to the cranium.

[0065] In aspects, the present disclosure provides a system for measuring microbubble concentration in the infusion liquid in real time, and in response adjust the infusion delivery rate to patients as to maintain optimal concentration of microbubbles in the patient blood throughout the duration of the treatment.

[0066] In aspects, the present disclosure provides a system for measuring microbubble concentration in the infusion liquid in real time based on infrared (IR) spectrophotometry. In someembodiments, the system comprises an apparatus capable of measuring absorbance of infrared (IR) light by the microbubbles in the infusion liquid. In some embodiments, the apparatus comprises two light emitting diodes (LEDs). In some embodiments, the two LEDs are positioned on the same horizontal line placed around a reservoir or below the reservoir. In some embodiments, the two LEDs are positioned at an angle that is between 0° and 360° with respect to each other around the center of the apparatus and / or around the reservoir. In some embodiments, the two LEDs are positioned at about 10°, or about 20°, or about 30°, or about 45°, or about 60°, or about 75°, or about 90°, or about 120°, or about 150°, or about 180° with respect to each other around the center of the apparatus and / or around the reservoir. In some embodiments, at the receiver LED is positioned at an angle different than 180° with relation to the emitting LED to measure light scattering. In some embodiments, more than one receiver LED is positioned at an angle different than 180° with relation to the emitting LED to measure light scattering. In other embodiments, a combination of one receiver LED positioned at the same line (180°) from the emitting LED and at least one receiver LED positioned at an angle different than 180° with relation to the emitting LED is used to measure a combination of light absorbed and scattered by the sample.

[0067] In some embodiments, the reservoir is drip chamber or infusion cup, right below the infusion bag. In some embodiments, one of the LEDs emits light in the IR spectrum through the infusion liquid accumulated in the cup, while the other LED measures how much light was absorbed by the accumulated liquid (FIG. 1A and FIG. IB). In some embodiments, the apparatus is connected to an infusion drip controller and can directly control the infusion drip rate based on the measured microbubble concentration. Alternatively, the apparatus can be connected to the transducer controller, which can control FUS parameters based on the measured microbubble concentration.

[0068] In aspects, the present disclosure provides a system that comprises a detector that measures light. In some embodiments, the detector uses at least one of spectrophotometry, light absorption or light transmission to measure a lipid-based ultrasound contrast agent concentration.

[0069] In aspects, the present disclosure provides a system capable of performing real-time microbubble concentration measurement during treatment directly on the infusion system.

[0070] In various embodiments, the system or methods of any of the embodiments disclosed herein measure microbubble ultrasound contrast agents concentration (for the sizing or / andcounting of microbubbles) is based either on IR spectrophotometry or simple light absorption or light transmission through the sample. In some embodiments, the microbubble concentration is determined in real time is used for patient treatment. In some embodiments, light absorption and / or light transmission can be used instead of light in the IR spectrum. In some embodiments, light in a specific wavelength or wavelength range may be used. In some embodiments, the microbubble concentration is usually linear with the absorption of the light passing through the sample even for light wavelengths that are not associated with specific components of the sample. For specific type of bubbles, the conversion parameters describes the linear relationship between the absorption and the concentration. The parameters can be found by building a calibration curve using measurements of samples with known concentration. An illustrative, non-limiting calibration curve may be found at FIG. 3.

[0071] In some cases the relation is not linear and there is a need in using several measurements of several concentrations spanning the range of interest of relevant concentrations to find the conversion parameters and to prepare an empirical conversion from measurement to concentration based on interpolation between the measurement points.

[0072] In some embodiments, the same apparatus can support measurements of several bubble types using different conversion parameters.

[0073] In various embodiments, the method disclosed herein is based on the measurement of the interactions between light and matter, and the reactions and measurements of radiation intensity and wavelength. Typical spectroscopy methods for use of certain vesicles use measurement of absorbance at 500 nm (OD50o) due to their proteic content. The present methods differ in that the microbubble ultrasound contrast agents are lipid based.

[0074] Measuring microbubble concentration in real time during treatment improves treatment safety and efficiency, by ensuring the delivery of optimal microbubble amount to the patient at all times during treatment, taking into consideration changes in microbubble concentration in the infusion liquid due to microbubble instability or other reasons.

[0075] Though microbubbles are the most commonly used contrast agents in FUS treatments, other contrast agents such as nanobubbles and phase shift droplets can also be used. Therefore, the term microbubble in this application includes to all types of contrast agents that can be used in FUS treatments.

[0076] The disclosure described here details a system that can be used to measure microbubble concentration in the infusion liquid in real time based on infrared (IR) spectrophotometry, and in response adjust the infusion delivery rate to patients as to maintain optimal concentration of microbubbles in the patient blood throughout the duration of the treatment.

[0077] In some embodiments, in addition to or as an alternative to measuring concentration of a composition, the concepts described herein can be used to measure the size and / or shape of the pharmaceutical constituents of the composition. The terms “pharmaceutical constituent” and “constituent” describe the elements (e.g., microbubbles or other therapeutic agents) that make up the composition. For example, measuring the light scattering can provide information regarding the size and shape of the microbubbles. In some embodiments, acoustic feedback can be used instead of light or IR. In some embodiments, acoustic waves in a specific wavelength range may be used. The acoustic wave wavelength may be changed depending on the properties (composition, density) of the molecules to be measured.

[0078] In some embodiments, some type of electro-magnetic wave source, like x-ray, may be used in a specific wavelength range. The electro-magnetic wave wavelength may be changed depending on the properties (composition, density) of the molecules to be measured.

[0079] In various embodiments, the method of treatment presented herein featuring the measurement of microbubble concentration, size, and / or shape in real time during treatment improves treatment safety and efficiency, by ensuring the delivery of optimal microbubble amount to the patient at all times during treatment, taking into consideration changes in microbubble concentration, size, and / or shape in the infusion liquid due to microbubble instability or other reasons.

[0080] In some embodiments, the disclosure could be used to measure microbubble concentration, size, and / or shape in the infusion liquid in real time, and in response adjust the sonication parameters of the transducer (power, sonication duration, pressure, intensity, frequency) in order to achieve the desired treatment effect (BBB opening level, BBB opening duration, vascular damage, drug release, neuro-modulatory effects including for psychiatric diseases such as addiction, depression, obsession compulsion disorder) or diagnostic effect (e.g. change echogenicity).

[0081] In another embodiment, the disclosure could be used to measure the concentration, size, and / or shape of a therapeutic in the infusion liquid in real time, and in response adjust the infusion delivery rate to patients as to maintain optimal concentration, size, and / or shape of therapeutics in the patient blood throughout the duration of the treatment.

[0082] In some embodiments, the composition comprises or further comprises a biologic drug selected from an antibody, a virus, a vaccine composition, a protein therapeutic, and a nucleic acid therapeutic selected from RNA, mRNA, chemically modified mRNA, a small interfering RNA (siRNA), a microRNA (miRNA), and an antisense RNA. In some embodiments, the composition comprises or further comprises a small molecule drug.

[0083] In addition to microbubbles, other therapeutic agents can be delivered to a patient intravenously during treatment. This is done, for example, in Focused Ultrasound therapies aimed at opening the blood-brain barrier (BBB) to facilitate drug delivery into the brain parenchyma. In some embodiments, the therapeutic agent comprises a small molecule or a biologic drug. In some embodiments, the therapeutic agent is or comprises a biologic drug. In some embodiments, the therapeutic agent is selected from a gene therapy agent, a vaccine, an antisense oligonucleotide (ASO), a protein therapeutic, a modified mRNA agent, and a RNAi agent. In some embodiments, the therapeutic agent is or comprises an antibody, antibody-like molecule or an antigen-binding fragment thereof. In some embodiments, the therapeutic agent specifically binds the protein (or another biomolecule) that exhibits abnormal production, aggregation, and / or deposition. In some embodiments, the therapeutic agent is selected from a nonspecific clearing antibody (e.g., intravenous immunoglobulin aka IVIg), an anti-amyloid-P antibody (e.g., aducanumab, gantenerumab, lecanemab, and donanemab), an anti-tau antibody (e.g., semorinemab, gosuranemab, tilavonemab, and zagotenemab), an anti-TREM2 antibody (e.g., AL002), an anti- alpha-synuclein antibody (e.g., Cinpanemab, Prasinezumab, Lu AF82422, ABBV-0805, and MEDI1341), and or a combination thereof. In some embodiments, the antibody is capable of binding a programmed cell death protein 1 (PD 1) (pembrolizumab (KEYTRUDA), nivolumab (OPDIVO), cemiplimab (LIBTAYO), pidilizumab (CT 011), dostarlimab (JEMPERLI), Spartalizumab (PDR001), RMP1-14, balstilimab (AGEN2034), Vopratelimab (JTX-4014), Camrelizumab (SHR1210), Sintilimab (IBI308), Tislelizumab (BGB-A317), Toripalimab (JS 001), INCMGA00012 (MGA012), AMP-224, and AMP-514), a PD 1 ligand (atezohzumab (TECENTRIQ), avelumab (BAVENCIO), and durvalumab (IMFINZI), KN035, and Cosibehmab(CK-301)), and / or cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) (Ipilimumab (YERVOY) or Tremelimumab (IMJUDO)). In some embodiments, the therapeutic agent is or comprises a small molecule drug. In some embodiments, the therapeutic agent provides one or more of synaptic plasticity, neuroprotection, reduction of inflammation, neurotransmitter receptor modulation, reduction of oxidative stress. 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 a combination thereof. In any of the embodiments disclosed herein, the therapeutic agent may be formulated in a liposome. In some embodiments, the therapeutic agent consists of a type of enzyme for enzyme replacement therapies. In some embodiments, the therapeutic agent consists of antibiotics and / or antivirals for brain infections. As with microbubbles, the concentration, size, and / or shape of these therapeutics is not measured in real-time during treatment, and hence, a solution to measure microbubble concentration, size, and / or shape in real time during treatment can also apply to measuring the concentration, size, and / or shape of other therapeutics delivered to the patient by infusion. These therapeutics can be used to treat tumors, neurodegenerative diseases, psychiatric disorders, genetic diseases and / or brain infections.

[0084] In some embodiments, the apparatus and / or the controller communicate with one or more additional piece of medical equipment. In some embodiments, the apparatus and / or the controller adjusts the desired concentration, size, and / or shape based on an input from the additional piece of medical equipment. In some embodiments, the apparatus and / or the controller provide an input to the additional piece of medical equipment to control its function.

[0085] In some embodiments, the additional piece of medical equipment is an ultrasound transducer device. In some embodiments, the additional piece of medical equipment is an imaging device. In some embodiments, the additional piece of medical equipment is selected from a computer tomography (CT) device, a magnetic resonance imaging (MRI) device, a positron emission tomography (PET) device, a single-photon emission computed tomography (SPECT) device, an ultrasonography device, a functional MRI (fMRI) device, an X-ray fluoroscopy device, a diffusion-tensor imaging (DTI) device, an angiography device (including a magnetic resonance angiography (MRA) device), a magnetoencephalography (MEG) device, anelectroencephalography (EEG) device, a near-infrared spectroscopy (NIRS) device, an optical imaging device, a bone densitometry device and a combination of two or more thereof.

[0086] In some embodiments, the device capable of detecting and localizing the one of more loci is employed to characterize tissue types and / or properties of the one or a plurality of regions and / or its surrounding tissue; each type and location of tissue, depending on its properties, may have corresponding tolerances for the acoustic response level and acoustic response. In some embodiments, the device may measure the cavitation effects (e.g., a temperature increase or an area that is disrupted) on the one or a plurality of regions and / or the surrounding tissue in real time. If an undesired effect on the one or a plurality of regions and / or its surrounding tissue is observed (e.g., the temperature increase exceeding a threshold and / or a disrupted area larger than a desired size), the ultrasound procedure may be halted. Accordingly, approaches described in the current disclosure may advantageously avoid permanent damage of the one or a plurality of regions and its surrounding tissue by reliably detecting microbubble cavitation events and monitoring effects of the cavitation on the target and / or surrounding tissue in real time.

[0087] In some embodiments, the controller is configured to cause the transducer to transmit acoustic energy to the target region at a transmission frequency, acquire a cumulative harmonic response from at least the target region, and operate the transducer based at least in part on the acquired cumulative harmonic response. For purposes hereof, operating based at least in part on the acquired cumulative harmonic response means based on the response from the target region, from around the target region, or from both target and non-target regions.

[0088] In some embodiments, the harmonic response is acquired at one or more positive integer multiples of the transmission frequency and / or at one or more positive off-integer multiples of the transmission frequency. In some embodiments, the system includes a filter for filtering the measured acoustic signals from the target region and / or its surrounding regions to obtain the cumulative harmonic response. In some embodiments, the filter is configured to select at least one of a harmonic, an ultraharmonic or a sub-harmonic response to the transmitted acoustic energy.

[0089] In some embodiments, the controller is further configured to compute the cumulative harmonic response by integrating a received acoustic signal from at least the target region over a predetermined time period. In some embodiments, the controller is configured to cause generation of microbubbles in the target region. In some embodiments, the system comprises anadministration device for introducing microbubbles into at least one target region and / or one or more surrounding region.

[0090] In some embodiments, temporarily altering a tissue characteristic comprises or consists of disrupting one or a plurality of regions. In some embodiments, the one or a plurality of regions are the BBB and the disruption may alter its permeability. In some embodiments, the controller is configured to control a parameter (such as power, frequency, pulse duration and / or pulse repetition frequency) of the transmitted acoustic energy based at least in part on spectral components of the cumulative harmonic response. In some embodiments, the controller is configured to control a parameter of the transmitted acoustic energy based at least in part on cumulative harmonic response data from within a defined interval. In some embodiments, the interval is within a current sonication. In some embodiments, the interval includes data from at least one previous sonication.

[0091] In some embodiments, the controller is configured to control a parameter to select for a harmonic frequency band while maintaining cumulative broadband emission and / or cumulative ultra-harmonics below corresponding safety thresholds. Alternatively, or in addition, the controller may be configured to control a parameter to increase the ratio between cumulative harmonics and cumulative ultra-harmonics, and / or between (i) cumulative harmonics and / or cumulative ultraharmonics and (ii) cumulative broadband emission.

[0092] Another aspect of the present disclosure provides a system comprising one or more computer processors and computer memory coupled thereto. The computer memory comprises machine executable code that, upon execution by the one or more computer processors, implements any of the methods above or elsewhere herein.

[0093] Another aspect of the present disclosure provides (i) a system for measuring concentration, constituent size, and / or constituent shape of a composition (without limitation, e.g., a microbubble composition) in real time at the time of a parenteral administration to a subject, (ii) an ultrasound transducer for sonicating a target volume to cause disruption of a target tissue therein system, and (iii) comprising one or more computer processors and computer memory coupled thereto. In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the one or more computer processors and computer memory coupled thereto comprise the controller of any of the embodiments disclosed herein. In some embodiments, the ultrasound transducer is capable of delivering a focused ultrasound (FUS). In some embodiments,the focused ultrasound (FUS) is selected from magnetic resonance imaging (MRI)-guided FUS, computerized tomography (CT)-guided FUS, electroencephalogram (EEG)-guided FUS, and positron emission tomography (PET)-guided FUS. In some embodiments, the FUS is MRI-guided FUS. In some embodiments, the computer memory comprises machine executable code that, upon execution by the one or more computer processors and / or controller, measures the concentration, constituent size, and / or constituent shape of the composition (without limitation, e.g., a microbubble composition) in real time at the time of a parenteral administration, adjusts the flow rate of the composition that is administered to the subject and / or provides input to the ultrasound transducer.

[0094] In some embodiments, the input from the apparatus and / or the controller controls the kind of an ultrasound beam (e.g., a low intensity focused ultrasound (LIFU) or a high intensity ultrasound (HIFU)), the center frequency of ultrasound beam (e.g., a specific value in the range of about 30 kHz to about 3 MHz, such as about 220 kHz), a shape of an ultrasound beam, a pulse pattern of an ultrasound beam, a spatial peak temporal average acoustic intensity (Ispta) (e.g., a specific value in the range of about 5 and about 1000 W / cm2), a spatial peak pulse average acoustic intensity (Isppa), a power of the ultrasound beam (e.g., a specific value in the range of about 0.5 W to about 100 W, e.g., about 5 W), and / or the duration of sonication (e.g., a specific value in the range of about 10 seconds and about 160 minutes such as about 60 seconds). In some embodiments, the input from the apparatus and / or the controller controls the kind of an ultrasound beam, a shape of an ultrasound beam, a pulse pattern of an ultrasound beam, the center frequency of ultrasound beam, a spatial peak temporal average acoustic intensity (Ispta), a spatial peak pulse average acoustic intensity (Isppa), a power of the ultrasound beam, and / or the duration of sonicationMethods for Measuring Concentration, Constituent Size, and / or Constituent Shape of a Composition and Methods of Treatment

[0095] In aspects, the present disclosure provides a method for measuring concentration, constituent size, and / or constituent shape of a composition in real time at the time of a parenteral administration, the method comprising: (a) providing a composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, (c) connecting the system to a port of administration of the composition to the subject, (d) measuring the physical property by the detector; and (e) determining the concentration, constituent size, and / or constituent shapeof the composition. In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0096] In some embodiments, the determining is performed using a standard curve of test measurements of the physical property and test concentrations, constituent sizes, and / or constituent shapes of the composition. In some embodiments, the standard curve is saved in the controller.

[0097] In some embodiments, the determining is performed using an equation. In some embodiments, the equation provides a correlation between test measurements and test concentrations, constituent sizes, and / or constituent shapes to determine the concentration, constituent size, and / or constituent shape. In some embodiments, the equation is saved in the controller.

[0098] In some embodiments, the physical property is selected from light absorbance, fluorescence and phosphorescence.

[0099] In aspects, the present disclosure provides a method of administering a composition to a subject. In some embodiments, the method comprises (a) providing a composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, and (c) connecting the system to a port of administration of the composition to the subject. In some embodiments, the composition is or comprises a microbubble composition. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0100] In aspects, the present disclosure provides a method of treating a subject in need thereof suffering from a disease or disorder. In some embodiments, the method comprises a step of administering a composition to the subject. In some embodiments, the step of administering comprises (a) providing the composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, and (c) connecting the system to a port of administration of the composition to the subject. In some embodiments, the method further comprises administering an ultrasound beam to the subject a second composition that is capable of preventing or treating the disease or disorder. In some embodiments, the composition is or comprises one or more microbubble compositions. In some embodiments, the one or more microbubble compositions are administered immediately before and / or during the application of the ultrasound beam. In some embodiments, the one or more microbubble compositions areadministered contemporaneously with the application of the ultrasound beam. In some embodiments, the composition is or comprises a microbubble compositions. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0101] In aspects, the present disclosure provides a method of treating a subject in need thereof suffering from a disease or disorder that is capable of being treated with ultrasound. In some embodiments, the method comprises a step of administering a composition. In some embodiments, the step of administering comprises (a) providing a composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, and (c) connecting the system to a port of administration of the composition to the subject. In some embodiments, the method further comprises administering an ultrasound beam to the subject. In some embodiments, the method further comprises administering an ultrasound beam to the subject and a second composition that is capable of preventing or treating the disease or disorder. In some embodiments, the composition is or comprises one or more microbubble compositions. In some embodiments, the one or more microbubble compositions are administered immediately before and / or during the application of the ultrasound beam. In some embodiments, the one or more microbubble compositions are administered contemporaneously with the application of the ultrasound beam. In some embodiments, the composition is or comprises a microbubble compositions. In some embodiments, the composition is or comprises a pharmaceutical composition.

[0102] In aspects, the present disclosure provides a method of diagnosing a disease or disorder in a subject. In some embodiments, the method comprises a step of administering a composition to the subject. In some embodiments, the step of administering comprises (a) providing the composition in a reservoir, (b) connecting the reservoir to the system of any of the embodiments disclosed herein, and (c) connecting the system to a port of administration of the composition to the subject. In some embodiments, the method further comprises administering an ultrasound beam to the subject and a second composition that is capable of preventing or treating the disease or disorder. In some embodiments, the composition is or comprises a pharmaceutical composition. In some embodiments, the composition is or comprises one or more microbubble compositions. In some embodiments, the one or more microbubble compositions are administered immediately before and / or during the application of the ultrasound beam. In some embodiments, the one or more microbubble compositions are administered contemporaneously with the application of the ultrasound beam. In some embodiments, the method further comprises obtaining a biologicalsample from the subject. In some embodiments, the biological sample is blood. In some embodiments, the method further comprises measuring the amount in the biological sample of a molecule (without limitation, a nucleic acid, a protein or a metabolite). In some embodiments, the molecule is predominantly present in an affected tissue but substantially not present in the biological sample under normal conditions. Such methods are disclosed in WO 2023 / 275617, the contents of which are incorporated by reference in their entirety.

[0103] In some embodiments, the composition comprising microbubbles is administered immediately before and / or during the application of the ultrasound beam. In some embodiments, the composition comprising microbubbles is administered contemporaneously with the application of the ultrasound beam.

[0104] In some embodiments, the disease or disorder is selected from a neurological condition. In some embodiments, the neurological condition is selected from Alzheimer’s Disease, Parkinson's Disease, essential tremor, neuropathic pain, epilepsy, addiction, post-traumatic stress disorder, depression and obsessive-compulsive disorder. In some embodiments, the disease or disorder is selected from a musculoskeletal pathology. In some embodiments, the musculoskeletal pathology is selected from osteoarthritis, soft tissue shoulder pathology, and myofascial pain. In some embodiments, the disease or disorder is selected from a cancer. In some embodiments, the cancer is a brain tumor or a metastatic brain tumor. In some embodiments, the disease or disorder is selected from a cardiovascular condition. In some embodiments, the cardiovascular condition is selected from varicose veins, blood clots, calcified arteries, and calcified veins. In some embodiments, the disease or disorder is selected from a kidney condition. In some embodiments, the kidney condition is a kidney stone. In some embodiments, the disease or disorder is selected from an eye condition. In some embodiments, the eye condition is selected from dry eye disease, glaucoma, presbyopia, macular degeneration, retinal impairment, and retinal injury. In some embodiments, wherein the second composition to the subject, wherein the second composition comprises a pharmaceutical agent that is used for treating the neurological condition, the musculoskeletal condition, the cancer, the cardiovascular condition, the kidney condition, or the eye condition.Kits

[0105] In some embodiments, present disclosure provides kits that can simplify the undertaking any method described herein. An illustrative kit of the disclosure comprises any composition described herein in a reservoir and / or a system described herein, e.g., a system for measuring the concentration, constituent size, and / or constituent shape of the composition. In some embodiments, the kit further comprises a device for applying an ultrasound beam across the cranium of the human patient (e.g., ultrasound transducers), and / or one or more microbubbles described herein in unit dosage form, and / or a positron emission tomography (PET) device, a single photon emission computed tomography (SPECT) device, an ultrasonography device, a diffusion-tensor imaging (DTI) device, a functional MRI (fMRI) device, a functional MRI (fMRI) device, an X-ray fluoroscopy device, an angiography device (including a magnetic resonance angiography (MRA) device), a magnetoencephalography (MEG) device, an electroencephalography (EEG) device, a near-infrared spectroscopy (NIRS) device, an optical imaging device, a bone densitometry device and a combination of two or more thereof.

[0106] In some embodiments, the unit dosage form is a container, such as a pre- filled syringe, or an infusion bag with gravitation-based flow controller, which can be sterile, containing any composition described herein and a pharmaceutically acceptable carrier, diluent, excipient, or vehicle.

[0107] The kit can further comprise a label or printed instructions instructing the use of any composition described herein.

[0108] As used herein, “microbubble” includes lipid based microbubbles (e.g., perflutren lipid microspheres), and microbubble alternatives, such as phase-shift droplets, nanobubbles, nanodroplets or nanoparticles, e.g., gold nanoparticles.EXAMPLES

[0109] The examples herein are provided to illustrate advantages and benefits of the present technology and to further assist a person of ordinary skill in the art with preparing or using the systems and apparatus for measuring drug concentration in real time according to the present technology. The examples herein are also presented in order to more fully illustrate the preferred aspects of the present technology. The examples should in no way be construed as limiting thescope of the present disclosure, as exemplified by the appended claims. The examples can include or incorporate any of the variations, aspects or embodiments of the present technology described above. The variations, aspects or embodiments described above may also further each include or incorporate the variations of any or all other variations, aspects or embodiments of the present technology.Example 1: Determination of Microbubble Concentration using an Infrared (IR) Device

[0110] Microbubble (MB) samples having different concentrations were prepared. The concentrations prepared were within the range of concentration of MBs that are clinically used in the infusion bags. MB concentrations were measured using a Counter coulter. MB concentrations were also assessed using an infrared device of the present disclosure. The infrared (IR) radiation generated by a light emitting diode (LED) was passed through the MB samples and the generated voltage was measured using an IR sensor LED. The generated voltage was plotted as a function of MB concentration, as measured using the Counter coulter. As shown in FIG. 3, there was a good correlation between the generated voltage with the MB concentration, as measured using the Counter coulter. Therefore, FIG. 3 serves a calibration curve for MB based on generated voltage.Example 2: Analysis Microbubble Stability Over Typical Clinical Treatment Time Frame

[0111] Microbubble (MB) samples having different concentrations, which were within the range of concentration of MBs that are clinically used in the infusion bags, were prepared by diluting a stock solution. MB concentrations in the dilutions were also assessed using an infrared device of the present disclosure at 0 min, 10 min, 20 min, 30 min, 40 min, 60 min, 80 min, 150 min, 180 min, 210 min, 240 min, and 270 min from preparation of the dilutions. The infrared (IR) radiation generated by a light emitting diode (LED) was passed through the MB samples and the generated voltage was measured using an IR sensor LED. The generated voltage was plotted as a function of time since the preparation of dilutions. The results are shown in FIG. 4. As shown in FIG. 4, a decrease in microbubble concentration was observed within the clinical treatment time frame (up to 4.5 hr). MBs concentration instability highlights the needs for a device that can measure concentration in real time, in order to ensure correct MBs concentration during the treatment and consequently, treatment efficiency and safety.Example 3: Analysis Microbubble Stability as a Function of Time and Concentrations

[0112] Microbubble (MB) samples having different concentrations were prepared by diluting a stock solution and divided in two groups (Group A and Group B) and aliquoted. Group A MB sample aliquots were assessed using an infrared device of the present disclosure after 20 min, 30 min, 40 min, 1 hr, 1.5 hr, 2 hr, 2.5 hr or 3 hr from preparation of the dilutions. Group A MB sample aliquots were assessed using an infrared device of the present disclosure after 3.5 hr, 4 hr, or 4.5 hr. Briefly, an infrared (IR) radiation generated by a light emitting diode (LED) was passed through the MB samples and the generated voltage was measured using an IR sensor LED. The generated voltage was plotted as a function of time since the preparation of dilutions. The results are shown in FIG. 5A. As shown in FIG. 5A, a decrease in microbubble concentration was observed within the clinical treatment time frame (up to 4.5 hr). The MB samples from Group A and Group B were also analyzed using a Coulter counter. As shown in FIG. 5B, Coulter counter measurement showed small changes in mean diameters of microbubbles. These results showed, inter alia, that microbubbles show instability both in their concentration and changes in diameter over a short time frame relevant for their clinical use.

[0113] MBs concentration instability highlights the needs for a device that can measure concentration in real time, in order to ensure correct MBs concentration during the treatment and consequently, treatment efficiency and safety.INCORPORATION BY REFERENCE

[0114] All patents and publications referenced herein are hereby incorporated by reference in their entireties.

[0115] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present technology is not entitled to antedate such publication by virtue of prior disclosure.

[0116] As used herein, all headings are simply for organization and are not intended to limit the disclosure in any manner. The content of any individual section may be equally applicable to all sections.EQUIVALENTS

[0117] While the disclosure has been disclosed in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the disclosure following, in general, the principles of the disclosure and including such departures from the present disclosure as come within known or customary practice within the art to which the disclosure pertains and as may be applied to the essential features hereinbefore set forth and as follows in the scope of the appended claims.

[0118] Those skilled in the art will recognize, or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments disclosed specifically herein. Such equivalents are intended to be encompassed in the scope of the following claims.

Claims

CLAIMSWhat is claimed is:

1. A system for measuring concentration, constituent size, or constituent shape of a composition in real time at the time of a parenteral administration, the system comprising:(a) an apparatus capable of measuring a physical property of the composition, the apparatus comprising a detector;(b) a controller, operably coupled to the detector, configured to:(i) store a desired concentration, constituent size, or constituent shape of the composition selected by an operator, optionally a human operator,(ii) measure the physical property of the composition, and(iii) adjust a flow rate of the composition in an outlet, optionally wherein the detector adjusts the flow rate based on a standard curve and / or an equation providing a correlation between test measurements and test concentrations, constituent sizes, or constituent shapes; and(c) the outlet being connected to a port of administration of the composition to a subject.

2. The system of claim 1 , wherein the detector adjusts the flow rate based on a standard curve.

3. The system of claim 1 or claim 2, wherein the physical property is selected from light absorbance, fluorescence and phosphorescence.

4. The system of any one of claims 1 to 3, wherein the apparatus comprises a light source and a detector of light, wherein the light source comprises a light emitting diode (LED), an infrared (IR) light, or an IR LED.

5. The system of any one of claims 1 to 4, wherein the detector provides an output of concentration, constituent size, or constituent shape of the composition.

6. The system of any one of claims 1 to 5, wherein the output is or comprises a print out and / or an on-screen display.

7. The system of any one of claims 1 to 6, wherein the reservoir of the composition is an infusion bag.

8. The system of any one of claims 1 to 7, wherein the reservoir of the composition is a drip chamber.

9. The system of any one of claims 1 to 8, wherein the parenteral administration is infusion.

10. The system of any one of claims 1 to 9, wherein the apparatus and / or the controller comprise a manual control unit configured to manually adjust the flow rate.

11. The system of any one of claims 1 to 10, wherein the flow rate is manually adjusted by a human operator, optionally based on the output of concentration, constituent size, or constituent shape of the composition.

12. The system of any one of claims 1 to 11, wherein the apparatus and / or the controller comprise an automated control unit configured to automatically adjust the flow rate.

13. The system of any one of claims 1 to 12, wherein the apparatus and / or the controller automatically adjust the flow rate based on the output of concentration, constituent size, or constituent shape of the composition and / or human input.

14. The system of any one of claims 1 to 13, wherein the apparatus and / or the controller automatically increase the flow rate compared to initial flow rate when the concentration, constituent size, or constituent shape of the composition is lesser compared to the desired concentration, constituent size, or constituent shape.

15. The system of any one of claims 1 to 14, wherein the apparatus and / or the controller automatically decrease the flow rate compared to initial flow rate when the concentration, constituent size, or constituent shape of the composition is greater compared to the desired concentration, constituent size, or constituent shape.

16. The system of any one of claims 1 to 15, wherein the apparatus and / or the controller automatically maintain the flow rate substantially equal to initial flow rate when the concentration,constituent size, or constituent shape of the composition is substantially equal compared to the desired concentration, constituent size, or constituent shape.

17. The system of any one of claims 1 to 16, wherein the composition is or comprises a microbubble composition.

18. The system of any one of claims 1 to 17, wherein the composition comprises or further comprises a biologic drug selected from an antibody, a virus, a vaccine composition, a protein therapeutic, and a nucleic acid therapeutic selected from RNA, mRNA, chemically modified mRNA, a small interfering RNA (siRNA), a microRNA (miRNA), and an antisense RNA.

19. The system of any one of claims 1 to 18, wherein the composition comprises or further comprises a small molecule drug.

20. The system of any one of claims 1 to 19, wherein the apparatus and / or the controller communicate with one or more additional piece of medical equipment, optionally selected from an ultrasound transducer, a positron emission tomography (PET) device, a single photon emission computed tomography (SPECT) device, an ultrasonography device, a diffusion-tensor imaging (DTI) device, a functional MRI (fMRI) device, a functional MRI (fMRI) device, an X-ray fluoroscopy device, an angiography device (including a magnetic resonance angiography (MRA) device), a magnetoencephalography (MEG) device, an electroencephalography (EEG) device, a near-infrared spectroscopy (NIRS) device, an optical imaging device, a bone densitometry device and a combination of two or more thereof.

21. The system of any one of claims 1 to 20, wherein the apparatus and / or the controller adjusts the desired concentration, constituent size, constituent shape and / or the flow rate based on an input from the additional piece of medical equipment.

22. The system of any one of claims 1 to 21 , wherein the apparatus and / or the controller adjusts the desired concentration, constituent size, constituent shape and / or the flow rate, based on an algorithm that analyzes the condition of the treated organ\body region.

23. The system of claim 22, wherein the condition of the treated organ\body region is provided to the controller by an operator, optionally a human operator.

24. The system of claim 22, wherein the condition of the treated organ\body region provided to the controller by the additional piece of medical equipment connected to the controller.

25. The system of any one of claims 1 to 24, wherein the apparatus and / or the controller provide an input to the additional piece of medical equipment to control its function.

26. The system of any one of claims 1 to 25, wherein the additional piece of medical equipment is an ultrasound transducer.

27. The system of claim 26, wherein the input from the apparatus and / or the controller controls the kind of an ultrasound beam, a center frequency of ultrasound beam, a power of the ultrasound beam, a shape of an ultrasound beam, a pulse pattern of an ultrasound beam, a spatial peak temporal average acoustic intensity (Ispta), a spatial peak pulse average acoustic intensity (Isppa), a frequency of ultrasound beam and / or a duration of sonication.

28. A method for measuring concentration of a composition in real time at the time of a parenteral administration, the method comprising:(a) providing a composition in a reservoir,(b) connecting the reservoir to the system of any one of claims 1 to 27,(c) connecting the system to a port of administration of the composition to the subject,(d) measuring the physical property by the detector; and(e) determining the concentration of the composition.

29. A method of administering a composition to a subject suffering from a disease or disorder, the method comprising(a) providing a composition in a reservoir,(b) connecting the reservoir to the system of any one of claims 1 to 27, and(c) connecting the system to a port of administration of the composition to the subject.

30. A method of treating a subject in need thereof suffering from a disease or disorder, the method comprising:(i) administering a composition comprising microbubbles in a reservoir to the subject, comprising(a) providing the composition,(b) connecting the reservoir to the system of any one of claims 1 to 27, and(c) connecting the system to a port of administration of the composition to the subject;(ii) administering an ultrasound beam to the subject; and(iii) optionally administering to the subject a second composition that is capable of preventing or treating the disease or disorder.

31. A method of treating a subject in need thereof suffering from a disease or disorder that is capable of being treated with ultrasound, the method comprising:(i) administering a composition comprising microbubbles in a reservoir to the subject, comprising(a) providing the composition,(b) connecting the reservoir to the system of any one of claims 1 to 27, and(c) connecting the system to a port of administration of the composition to the subject;(ii) administering an ultrasound beam to the subject; and(iii) optionally administering to the subject a second composition that is capable of preventing or treating the disease or disorder.

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