Field of the Invention

The invention relates to devices and methods for ultrasonic delivery of an agent to an internal tissue.

Background

The most common route of drug delivery is oral administration. Many drugs can be readily absorbed in the gastrointestinal (GI) tract, so oral administration allows them to enter the blood quickly and circulate systemically. In addition, oral administration is convenient and minimally invasive. Nonetheless, oral administration is not suitable for all drugs. For some drugs, the acidic conditions and harsh digestive enzymes of the GI tract degrade or inactivate the active pharmaceutical ingredient (API) before it can reach its target tissue. Other therapeutic agents, such as biological therapeutics ("biologies"), which generally consist of large macromolecules, are poorly absorbed in the GI tract. Absorption may also be limited if the patient has a diarrhea, which minimizes the duration of transit of the drug through the GI tract.

Ingestible ultrasonic drug delivery devices or capsules have been developed to overcome the difficulty of delivering certain drugs via the GI tract. Such devices incorporate the use of an ultrasound transducer, a reservoir that stores the drug, and a power source, such as a battery and drive circuitry, which drives the transducer. However, the utility of these fully self-contained devices is limited by a different set of technical obstacles. For example, the device must be small enough that it can be easily swallowed, yet large enough to accommodate the drug, transducer, drive circuitry, and battery. Conventional capsule electronics are highly volume inefficient, generally requiring multiple chips, packaging, and wires. The physical dimensions and mechanical characteristics of the device also dictate its biocompatibility with the gastrointestinal tract. For instance, the maximum size of a capsule’s rigid outer body is limited to the diameter of the smallest passage within the gastrointestinal tract. In addition, silver-oxide button batteries occupy significant real estate volume within ingestible capsules and often become the deciding factor for the device size. These factors constrain the quantity of drug that can be delivered by all-in-one ingestible ultrasonic drug delivery devices. Another consideration is that the battery can severely damage internal tissue if it makes electrical contact with the tissue. The alkaline solution within ingested silver-oxide button batteries can cause severe tissue damage in the mouth, vocal cord, trachea, or esophagus. The device must contain material to electrically insulate the battery, which further restricts the drug-loading capacity of the device. In addition, conventional methods for the mechanical fabrication of a capsule can limit the potential to reduce their volume. Consequently, these factors largely limit the therapeutic potential of drug delivery via ingestible ultrasonic devices.

Summary

Aspects of the present disclosure may include a low power high-volume efficient and integrated ingestible capsule drug delivery system for targeted or localized ultrasound-mediated drug delivery within the GI tract. The integrated ingestible capsule drug delivery system may comprise one or more energy storage device, magnetoelectric or magnetoelastic composite transducer, magnetoelectric (ME) transducer driver, magnetoelectric pH sensitive mass sensor, drug carrier/reser- voir, and at least one drug payload. In various embodiments, the ME or magnetoelastic composite transducer may comprise one or more layers of a flexible thin film magnetostrictive and a piezoelectric material coupled to transduce magnetic field energy to electric energy to produce mechanical vibrations generating low frequency ultrasound in a small form factor. In various embodiments, the one more layer of magnetoelastic material may comprise an amorphous ferromagnetic alloy and at least one layer of piezoelectric material. In various preferred embodiments, the amorphous alloy may comprise Fe4oNi4oPi4Be (Metglas) or the like and the piezoelectric material may comprise Polyvinylidene Fluoride (PVDF), Lead Zirconate titanate (PZT), or the like. In various embodiments, at least one magnetostrictive and piezoelectric layer are bonded together by an interfacial adhesive layer to enable mechanical coupling between the layers. In various embodiments, the ME transducer driver may comprise a magnetic field or flux generator further comprising a driver circuit, an H-bridge, and a series capacitor, said elements operating in conjunction with at least one magnetic coil, preferable a planar coil, and a bias DC magnetic field generator. In various embodiments, the ME pH sensitive mass sensor may comprise the said magnetic field generator, a magnetostrictive ribbon/strip or ME transducer, a magnetic field signal detection-receiver coil, and a pH sensitive polymer coating. In various embodiments, the ME transducer driver and ME mass pH sensor electronics are implemented using one or more IC or ASIC. In various embodiments, at least one energy storage device (e g., biocompatible supercapacitor, etc.), magnetoelectric or magnetoelastic composite transducer, ME transducer driver, ME mas pH sensor, drug carrier/reservoir, and drug payload are deposited, each in a defined zone, on a thin, flexible, flexible to rigid (F2R), or planar polymeric substrate, with one or more layers combined to function as a transducer, a driver, or sensor, in a specified area. In various embodiments, at least one drug is deposited within one or more drug carrier/reservoir and capped with one or more layer of a pH sensitive polymer. In various embodiments, the top layer of the whole substrate is coated with a pH sensitive polymeric layer and function as an integral element of the magnetostrictive or ME pH sensitive mass sensor. The final coated substrate is foldable at one or more bendable junction, substrate bended to form a cylindrical capsule whereby one or more bended proximal and distal end portions are inserted or positioned internally within the lumen of the folded substrate enabling the fabrication a cylindrical capsule with the electronic portion or components (i.e., battery, transducer electronics, etc.) contained and sealed within the internal lumen of the capsule to form an integrated low power high-volume efficiency ingestible low frequency ultrasound producing drug delivery system.

Aspects of the present disclosure may include methods for fabricating a low power high- volume efficient integrated ingestible capsule drug delivery system for targeted or localized ultra- sound-mediated drug delivery within the GI tract. In various embodiments, a method may comprise steps for coating or printing one or more layer of a thin flexible polymer substrate to produce one or more components of the drug delivery system, including but not limited to, an energy storage device, an ME transducer, an ME driver and or pH sensor, IC or ASIC, drug carrier/reservoir, and drug payload. In various embodiments, another method may comprise steps for encapsulating the drug delivery system for biocompatibility or dissolution of one or more protective or sustained drug release layer. In various embodiments, yet another method may comprise one or more fabrication steps resulting in a cylindrical capsule or thin rectangular tablet with electronic components and the energy storage device contained within the internal lumen or volume and sealed for waterproof. In various embodiments, the fabrication methods are combined to produce an integrated capsule drug delivery system capable of targeted delivery within the gastrointestinal (GI) tract, producing low frequency ultrasound to generate, including but not limited to, an ultrasound motive force, an ultrasound field gradient, a sonophoretic force, acoustic streaming, or cavitation incorporating one or more ME transducer and a magnetostatic or ME pH sensitive mass sensor. In various embodiments, one or more ME transducers are fabricated to produce low frequency ultrasound from one or more combination of variable parameters, including but not limited to, magnetostrictive and piezoelectric material, layer, length, width, height, length to width ratio, area ratio, laminate cross-sectional areas, interfacial coupling layer Young’s moduli, the like, or combinations thereof. The printing, coating, or additive manufacturing process may comprise the use of screen, inkjet, fl exo, gravure, the like, or combinations thereof.

Aspects of the present disclosure may include methods for targeted or localized ultrasound- mediated drug delivery within the GI tract using an ME transducer, ME pH sensitive mass sensor, a pH sensitive coated ingestible integrated capsule drug delivery system. In various embodiments, one method for targeted drug delivery may comprise determining a mass change of the drug delivery device or a portion of the device caused by a dissolution of a pH sensitive polymer encapsulating at least a portion or the whole/total of the surface areas of the device or a pH sensitive layer of a coating at least a portion or whole/total of the surface areas of the device. In various embodiments, another method may comprise the synchronous or asynchronous, continuous, or intermittent monitoring and detection of a shift in the electro-mechanical resonance of an ME sensor due a change in the mass or whole or portion of the drug delivery device. Upon detection of a specific change in device whole or portion mass, the ME transducer is actuated to produce one or more timed ultrasonic vibration to release at least one drug from the drug carrier/reservoir and subsequent active transport of the drug to, into, or within a GI cell, membrane, or tissue. In another aspect, yet another method may comprise administering a therapeutic agent to a GI tissue of a subject by transporting the ingestible capsule to at least one specific location of the GI and the payload containing an encapsulated or non-encapsulated therapeutic agent is activated by an ME transducer within the capsule for control-released, pulsatile, non-pulsatile, intermittent, digital, or continuous local or targeted delivery of said agent from the payload or reservoir into GI tissue of the subject. In another aspect, said drug reservoir may be configured to releasably retain at least one encapsulated therapeutic agent. In various embodiments, the ultrasound transducer may be positioned to transduce ultrasound waves in a particular direction relative to the reservoir of the ingestible capsule. The ultrasound transducer may be positioned to transduce ultrasound waves toward the reservoir. The ultrasound transducer may be positioned to transduce ultrasound waves away from the reservoir. The ultrasound transducer may be positioned to produce omnidirectional ultrasound waves through the reservoir. The reservoir may be configured to releasably retain a liquid comprising a therapeutic agent or encapsulated therapeutic agent. The resulting methods enable the implementation of a miniature low power high-volume efficient integrated ingestible capsule pH-dependent GI tract and or targeted ileocolonic drug delivery system.

The ME transducer of the integrated ingestible capsule may produce an ultrasound signal with a defined frequency or within a defined frequency range. The ME transducer may produce an ultrasound signal of from about 10 kHz to about 10 MHz, from about 10 kHz to about 1 MHz, from about 10 kHz to about 100 kHz, from about 20 kHz to about 80 kHz, from about 20 kHz to about 60 kHz, or from about 30 kHz to about 50 kHz. The ME transducer may produce an ultrasound signal of less than 100 kHz, less than 80 kHz, less than 60 kHz, or less than 50 kHz. The ME transducer may produce an ultrasound signal of about 20 kHz, about 25 kHz, about 30 kHz, about 35 kHz, about 40 kHz, about 45 kHz, about 50 kHz, about 55 kHz, or about 60 kHz. In various embodiments, the ME transducer may comprise at least one, directional, planar, spherical, hemi-spherical, or omni-directional transducer. The ingestible capsule may have a defined size or length. The ingestible capsule may have the longest dimension of less than about 3.0 cm, about 2.75 cm, about 2.5 cm, about 2.25 cm, about 2.0 cm, about 1.75 cm, or about 1.5 cm. The ingestible capsule may have a transverse dimension of less than about 1.2 cm, about 1.1 cm, about 1.0 cm, about 0.9 cm, or about 0.8 cm.

In another aspect, the present disclosure provides systems and methods of administering a therapeutic agent to a GI tissue of a subject by orally administering to a subject a low power high- volume efficient ingestible integrated capsule comprising a drug carrier/reservoir and a drug payload. In various embodiments, the drug payload may contain a therapeutic agent that is encapsulated in at least one pH, thermal, electric, magnetic, electromagnetic wave, catalytic, piezo-catalytic, or ultrasound-responsive polymeric carrier, including but not limited to, microbubble, nanobubble, nanodroplet, nano emulsion, nanofiber, vesicle, micelle, or hydrogel sphere or coating. In various embodiments, the ingestible capsule may comprise a coating, one or more pH sensitive coating layer or scaffold on at least one internal or external surface, said coating or scaffold contains at least one therapeutic agent encapsulated in at least one pH, thermal, electric, magnetic, electromagnetic wave, catalytic, piezo-catalytic, or ultrasound-responsive polymeric carrier. In various embodiments, the ingestible capsule may incorporate a drug carrier/reservoir containing an iron oxide particle-based biocompatible gel or microporous gel with a controlled architecture that can release its payload, containing said encapsulated or non-encapsulated therapeutic agent, when exposed to AC magnetic field or ultrasound vibration produced by at least one ME transducer of the present disclosure. In various embodiments, the said biocompatible gel or microporous gel containing at least one therapeutic agent may be deposited within one or more defined drug car- rier/reservoir using, including but not limited to, at least one additive manufacturing technique.

Brief Description of the Drawings

FIG. 1 is a pictorial of the various expanded layers of an integrated ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract, according to an embodiment of the present disclosure.

FIG. 2 is a diagram of a planar top view of the flexible integrated ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract, according to an embodiment of the present disclosure.

FIG. 3 is an illustration of an integrated ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract, according to an embodiment of the present disclosure.

FIG. 4 is a diagram of a magnetoelectric transducer driver and resonant detect or/recei ver integrated within the ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract, according to an embodiment of the present disclosure.

FIG. 5 is a flow chart of a method using ME mass sensing for targeted drug delivery using a low power high-volume efficient ingestible capsule is shown, according to an embodiment of the present disclosure.

Detailed Description

It should be appreciated that all combinations of the concepts discussed in greater detail below (provided such concepts are not mutually inconsistent) are contemplated as being part of the inventive subject matter disclosed herein. It also should be appreciated that terminology explicitly employed herein that also may appear in any disclosure incorporated by reference should be accorded a meaning most consistent with the concepts disclosed herein.

It should be appreciated that various concepts introduced above and discussed in greater detail below may be implemented in any of numerous ways, as the disclosed concepts are not limited to any particular manner of implementation. Examples of specific implementations and applications are provided primarily for illustrative purposes. The present disclosure should in no way be limited to the exemplary implementation and techniques illustrated in the drawings and described below.

Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed by the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed by the invention, subject to any specifically excluded limit in a stated range. Where a stated range includes one or both endpoint limits, ranges excluding either or both of those included endpoints are also included in the scope of the invention.

As used herein, the term “includes” means includes but is not limited to, the term “including” means including but not limited to. The term “based on” means based at least in part on.

As used herein, the term “volume efficiency” means the amount of function per unit volume displaced by a capsule or device.

Ultrasound- Mediated Drug Delivery System and Components

Aspects of the present disclosure can include a low power high-volume efficient integrated ingestible capsule drug delivery system for targeted or localized ultrasound-mediated drug delivery within the GI tract. The high-volume efficiency low power ingestible capsule drug delivery system may comprise one or more low power energy storage device, magnetic field or flux generator, a bias magnetic field component, a magnetoelectric (ME) or magnetoelastic transducer, ME transducer driver, drug carrier/reservoir, at least one drug payload, a pH sensitive polymer coating, and optionally a diagnostic unit. The diagnostic unit further may comprise an ME pH sensitive mass sensor operating in conjunction with a pH sensitive polymer outer dissolvable coating of the capsule to determine the location of the capsule during transit through the GI tract and for subsequent activation of the ME ultrasound transducer to an ultrasound motive force, ultrasound field gradient, sonophoretic force, acoustic streaming, or cavitation within the GI using low frequency ultrasound at a targeted location. The capsule may be fabricated by coating or depositing on a planar flexible substrate one or more layers of flexible alloys, materials, or polymers, including but not limited to, a conductive, magnetostrictive, magnetoelastic, piezoelectric, dielectric, adhesive, therapeutic drug payload, and a pH sensitive polymer coating. The resultant flexible substrate is foldable at one or more bendable junction, substrate bended to form a cylindrical capsule whereby one or more bended proximal and distal end portions are inserted or positioned internally into the lumen of the folded substrate enabling the fabrication a cylindrical capsule with the electronic portion or components (i.e., battery, transducer electronics, etc.) contained and sealed within the internal lumen of the capsule to form an integrated low power high-volume efficiency miniature ingestible low frequency ultrasound-mediated drug delivery system.

Referring now to FIG. 1, a pictorial 100 of the various expanded layers of an integrated ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract is shown, according to various embodiments. In various embodiments, the low power high-volume efficiency ingestible capsule drug delivery system 102 may comprise a flexible polymeric substrate 104 upon which the surface is coated or deposited with one or more thin component layers, at a specified portion or substantially throughout at least one surface of the substrate. In various embodiments, flexible polymeric substrate 104 can be any substrate employed in printed electronics, including but not limited to, thermal plastic polymer, PET, PI, PEN, PTU, or silicon elastic polymer, PDMS. In various embodiments, the component layers may comprise a direct- current (DC) bias magnetic field producer 106, a dielectric separation layer 108, a magnetic coil layer containing a driver coil 110 and a sensing coil 112, a magnetoelectric (ME) transducer layer 114, a drug payload layer 116, and a pH sensitive polymer coating layer 118. In various embodiments, flexible polymeric substrate 104 may comprise a proximal end portion 120 that is bendable at junction 122 and a distal end portion 124 that is bendable at junction 126. In various embodiments, distal end portion 124 may comprise the deposition and or placement of an energy storage device 128 and one or more integrated circuit (IC) or application specific IC (ASIC) device 130. In various embodiments, energy storage device 128, which may comprise a miniature battery or supercapacitor, preferably biocompatible, provides energy to power device 130 for operating an ME transducer made up of layer 114 as well as an ME pH-sensitive mass sensor made up of layers 114, 116 and 118. In various embodiments, the energy storage device 128 may comprise a biocompatible battery or a biocompatible capacitor, a biocompatible supercapacitor, combinations thereof, or the like. In various embodiments, said biocompatible energy devices may incorporate one or more anode, cathode, and electrolytes to be biodegradable or safe for ingestion. In various embodiments, the anode-cathode-electrolyte combinations can be but not limited to: Mg/Fe/PCL/NaCl composite activated H2O, Mg/CuCl/SGF, Zn-Mg/Cu/SGF, Zn/SGF, AC- XMnO2/H20 1 M Na2SO4, or Melanin/ XMnC 1 M Na2SO4. In various embodiments, energy storage device 128 may be a battery encapsulated for compatibility using, for example, Parylene deposited as a thin fdm. In an alternative embodiment, the ME pH sensitive mass sensor may comprise a strip layer of magnetostrictive alloy strip (e.g., Metglas) deposited between layer 116 and layer 118 to directly sense mass changes resulting from the dissolution of layer 118 as a function of GI pH. In various embodiments, the bias magnetic field producer comprises a ferrite layer, preferable printed with ferromagnetic or rare earth material, for example Neodymium Iron Boron. In various embodiments, the ME or magnetoelastic composite transducer may comprise one or more layers of a flexible thin film magnetostrictive and a piezoelectric material that are coupled to transduce magnetic field energy to electric energy to produce mechanical vibrations generating low frequency ultrasound in a small form factor. In various embodiments, the one more layer of magnetostrictive material may comprise an amorphous ferromagnetic alloy and at least one layer of piezoelectric material. In various preferred embodiments, the amorphous alloy may comprise Fe4oNi4oPi4Be (Metglas) or the like and the piezoelectric material may comprise Polyvinylidene Fluoride (PVDF), Lead Zirconate titanate (PZT), or the like. In alternative embodiments, the ME transducer may comprise a combination of magnetostrictive and piezoelectric materials including but not limited to Metglas/PVDF, Fe-Co alloy s(Ni)/PZT, FeCuNbSiB/PZT, or FeCoSiB/Alumi- num Nitrite. In yet other alternative embodiments, the ME pH-sensitive mass sensor may include crystalline alloys (e.g., 50 Co, 50 Fe; 50Ni, 50 Ni; 97 Fe, 3 Si, Ni, TbDyFe2) or amorphous alloys (FsoBuSie; COvsSiisBio; COesNiioBuSis) or Terfenol, or Galfenol, the like, or combinations thereof. In various embodiments, at least one magnetostrictive and piezoelectric layer are bonded together by an interfacial adhesive layer, for example epoxy, to enable mechanical coupling between the layers. In various embodiments, the ME transducer driver may comprise a bias magnetic field produced by producer 106 or flux generator further comprising a driver circuit, an H-bridge, and a series capacitor, said elements operating in conjunction with at least one magnetic coil 110, preferable planar, and bias DC magnetic field generator or producer 106. In various embodiments, the ME pH sensitive mass sensor may comprise the said magnetic field generator 106, a magnetostrictive ribbon/ strip or ME transducer of layer 114, a magnetic field signal detection-receiver coil 112, and a pH sensitive polymer coating layer 118. In various embodiments, the ME transducer driver ME and mass pH sensor electronics are implemented using ASIC 130. In various embodiments, energy storage device 128, magnetoelectric or magnetoelastic composite transducer 114, ME transducer driver, ME pH sensitive mass sensor made up of layers 114, 116 and 118, drug carrier/reservoir 116, and drug payload are deposited, each in a defined zone, a thin, flexible, flexible to rigid (F2R), planar polymeric substrate 104, with one or more said layers combined function as a transducer, a driver, or sensor, in a specified portion or substantially whole surface area of substrate 104. In various embodiments, at least one drug is deposited within one or more drug carrier/reservoir located within layer 116 and capped with at least one pH sensitive polymer layer 118. In various embodiments, the top layer of the whole substrate 104 is coated with pH sensitive polymeric layer 118 and function as integral element of the magnetostrictive or ME pH sensitive mass sensor. In various embodiments, each said layers deposited using one or more printing technology as to enable intimate contacts between the layers resulting an integrated planar structure. The final coated planar substrate 104 is then foldable at bendable junctions 112,116, said substrate bended to form a cylindrical capsule whereby bended proximal 122 and distal 126 end portions are inserted or positioned internally within the lumen of the folded substrate 104 enabling the fabrication a cylindrical capsule with the electronic portion or components (i.e., device 124 and device 130) contained and sealed within the internal lumen of the capsule to form an integrated low power high-volume efficiency ingestible low frequency ultrasound producing drug delivery system.

An aspect of the present disclosure is the fabrication of an ME transducer as an integral component of an ingestible capsule for ultrasound-mediate drug deliver. The ME transducer of the integrated ingestible capsule may produce an ultrasound signal with a defined frequency or within a defined frequency range. In various embodiments, one or more ME transducers are fabricated to produce low frequency ultrasound from one or more combination of variable parameters, including but not limited to, magnetostrictive and piezoelectric material, layer, length, width, height, length to width ratio, area ratio, laminate cross-sectional areas, interfacial coupling layer/factor, interfacial coupling/bonding material’s Young’s moduli, strength of DC bias magnetic field, the like, or combination thereof. In various embodiments, the resonant frequency of the ME transducer may be adjusted to a desired operating frequency by varying the length of the magnetostrictive and piezoelectric layer, the interfacial coupling factor, or their thickness ratio. In one embodiment, the ME transducer film length may be in the non-limiting range of 1 to 25 mm with a resonant frequency within 50 to 500 kHz. In various embodiments, low frequency may be generated with using a non-limiting low amplitude magnetic field (e.g.,< 10 mTesla). It has been discovered by others that the ME voltage coefficient is independent of area of the laminated film and increases or decrease depending on the interface coupling factor. In one embodiment, the ME transducer may be fabricated with a non-limiting fdm thickness of 10 to 50 pm of Metglas glued using epoxy to a non-limiting 100 to 300 pm thick layer of PZT, preferably ratios with the piezoelectric layer being thicker than the magnetostrictive layer to achieve low frequency ultrasound. In another embodiment, the ME transducer may be fabricated with a non-limiting 25 to 150 mm thick layer of PZT to increase vibrational fdm deflection when the applied magnetic field frequency is matched to the ultrasound resonant frequency. In various embodiments, PVDF may serve as an alternative to PZT. In various embodiments, the thickness ratio of the magnetostrictive and piezoelectric layers is between non-limiting 3 to 6. In various embodiments, the ME transducer or ME pH sensitive mass sensor may be fabricated to align one or more of their vibrational nodes (preferably locations of minimum or zero displacement) with segments of coil 110 or coil 112 of FIG. 1 for support whereby the one or more aligned specific coil segments have a thickness fractionally greater than others. Depending on these variables, the ME transducer may be fabricated to produce an ultrasound signal of from about 10 kHz to about 10 MHz, from about 10 kHz to about 1 MHz, from about 10 kHz to about 100 kHz, from about 20 kHz to about 80 kHz, from about 20 kHz to about 60 kHz, or from about 30 kHz to about 50 kHz. The ME transducer may produce an ultrasound signal of less than 100 kHz, less than 80 kHz, less than 60 kHz, or less than 50 kHz. The ME transducer may produce an ultrasound signal of about 20 kHz, about 25 kHz, about 30 kHz, about 35 kHz, about 40 kHz, about 45 kHz, about 50 kHz, about 55 kHz, or about 60 kHz. In various embodiments, the ME transducer may comprise at least one, directional, planar, spherical, hemispherical, or omni-directional transducer. In various embodiments, the ME transducer may be tailored by stressing the sensor in a controlled fashion, either through in-elastic dimpling of the transducer or elastically bending it. The inherent stress causes the ME transducer to vibrate out-of-plane to produce larger vibrational displacement. The ingestible capsule may have a defined size or length based on the populated components. The ingestible capsule may have the longest dimension of less than about 3.0 cm, about 2.75 cm, about 2.5 cm, about 2.25 cm, about 2.0 cm, about 1.75 cm, or about 1.5 cm. The ingestible capsule may have a transverse dimension of less than about 1.2 cm, about 1.1 cm, about 1.0 cm, about 0.9 cm, or about 0.8 cm.

Referring now to FIG. 2, a diagram 200 of the planar top view of the flexible integrated ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract is shown, according to various embodiments. The flexible integrated ingestible capsule drug delivery system comprises one or more specified area populated with one or more system components deposited on a flexible substrate 202 which is equivalent to substrate 102 of FIG. 1. In various embodiments, an exemplary primary area 204 comprises a stack of layers making up, including but not limited to, the ME transducer, drug payload, ME pH sensitive mass sensor, and pH sensitive dissolvable coating of the ingestible capsule. In various embodiments, primary area 204 is deposited or coated with a first DC bias magnetic field generator layer 206 (equivalent to layer 106 of FIG. 1), a second magnetic coil driver and receiver layer 208 (equivalent to layer containing microcoils 110 and 112 of FIG. 1), a third ME transducer layer 210 (equivalent to layer 114 of FIG. 1), a fourth drug payload layer 212 (equivalent to layer 116 of FIG. 1), and a final pH sensitive outer coating layer 214(equivalent to layer 118 of FIG. 1). In various embodiment, another exemplary area 216, equivalent to the top surface of distal end portion 124 of FIG. 1, may be populated with an energy storage device 218, equivalent to energy storage device 128 of FIG. 1 and ASIC 220, equivalent to ASIC 130 of FIG. 1. In an analogous manner, exemplary area 222, equivalent to the top surface of proximal end portion 120 of FIG. 1 may or may not be populated but service as an end cap for the ingestible when folded an inserted or position internal to the capsule. In various embodiments, the flexible substrate 202 with one or more defined deposition areas a cut or shape to enable the substrate to be folded into a cylindrical capsule whereby edge 224 of primary area 204 is folded or rolled substantially to create an inner core or scaffold and edge 226 is secured, using adhesive or the like, to an external body portion of the resulting rectangular or cylinder of the to-be-formed capsule. In various embodiments, portions 222 and 216 are first folded and then edge 224 is folded or rolled to encapsulate portion 216 and portion 222 of flexible substrate 202. In various alternative embodiments, primary area 204 of flexible substrate 202 may be wrapped around a rectangular scaffold or cylindrical scaffold, of approximate length, width, or diameter, with a proximal and distal receptacle for insertion of portion 222 and portion 216, respectively, to form a capsule having the coated layers on the exterior side. In various embodiments, any exposed external surface of the capsule may be further coated with pH sensitive polymer layer 214 via coat dipping to produce a completely pH sensitive capsule. In various embodiments, pH sensitive polymer layer 214 may comprise one or more pH sensitive polymer, including but not limited to, cellulose acetate phthalates (CAP), hydroxypropyl methyl-cellulose phthalate (HPMCP) 50 and 55, copolymers of methacrylic acid and methyl methacrylate (e.g., Eudragit® S 100, Eudragit® L, Eudragit® FS, and Eudragit® P4135 F). In various embodiments, the pH sensitive polymer may incorporate the ColoPulse system in which a disintegrant, such as sodium starch glycolate or croscarmellose sodium, is incorporated into a pH-sensitive polymeric coating layer in a non-percolating manner to prevent premature dissolution. It is understood that the said layers deposited in primary area 204 are coated and aligned with substantial equivalent overlapping surface area and dimensions. Diagram 200 is drawn only for illustrative purposes and is understood to be non-limiting.

Referring now to FIG. 3, an illustration 300 of an integrated ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract is shown, according to various embodiments. The ingestible capsule drug delivery system may comprise a cylindrical portion 302 made up of flexible substrate 304, equivalent to flexible substrate 202 of FIG. 2, said substrate populated with system components produced by one or more coating layers. Cylindrical portion 303 may be formed as described in FIG. 2 to produce an internal lumen 306 to accommodate, planar portion 308, equivalent for example to distal portion 216 of FIG. 2, positioned internally, and shown to be populated with ASIC 310, equivalent to ASIC 220 of FIG. 2, for controlling the operation of the capsule, powered by energy storage device 312, equivalent to 218 of FIG. 2. In various alternative embodiments, planar portion 308 may be further supported inside lumen 306 with an optional scaffold 314. In various exemplary embodiments, the flexible substrate 304 is coated with one or more layer of system components, including but not limited to, a ME magnetic driver -receiver coil layer 316, equivalent to layer 208 of FIG. 2, an ME transducer layer 318, equivalent to layer 210 of FIG. 2, and a pH sensitive coating layer 320, equivalent to layer 214 of FIG. 2. It is understood that the said layers deposited on flexible substrate 304 are coated and aligned with substantial equivalent overlapping surface area and dimensions. The printing, coating, or additive manufacturing process may comprise the use of screen, inkjet, fl exo, gravure, the like, or combination thereof. The illustration 300 is drawn only for explanatory purposes and is understood to be non-limiting. The resulting structure is a low power high-volume efficient integrated ingestible capsule drug delivery system afforded using additive manufacturing to produce miniature, easily swallowable, and amenable to large quantity production.

Referring now to FIG. 4, a diagram 400 of a magnetoelectric transducer driver and resonant detector/receiver integrated within the ingestible capsule drug delivery system for targeted ultrasound-mediated drug delivery within the GI tract is shown, according to various embodiments. The ME transducer driver 402 may comprise a logic module 404 electrically connected to a bridge driver 406 further electrically connected to an H-bridge driver 408, used to excite planar coil 410, equivalent to coil driver coil 110 of FIG. 1. In various embodiments, ME transducer driver 402 may be activated to produce low frequency ultrasound, at a programmed power, intensity, frequency, or duty cycle to disperse one or more drug payload from one or more drug carrier/reservoir layer 116 of FIG. 1. In various embodiments, ME transducer driver 402 may operate in combination with ME mass sensor detection module 412 to produce low ultrasound to control release drug payload and subsequent active transport of said release drug into GI tissue at one or more predetermined location within the GI tract. In various embodiments, module 412 receives one or more detected signal from sensor coil 414, equivalent to coil 112 of FIG. 1, which is electrically connected optionally to a pre-amp 416, said pre-amp condition the detected signals for processing by a resonant detector 418, electrically connected to a logic control module 420. In various embodiments, the functions performed by module 420 may be performed by module 404 or vice versa, to conserve real estate for high-volume efficiency. In various embodiments, driver 402 and detector module 412 operate together with layers 114, 116, 118 of FIG. 1 as an ME pH sensitive mass sensor of the present disclosure. Without being bound to theory, in general, the principle of operation is based on the use of the mechanical vibration of the ME sensor generated through the magnetoelastic effect by sending a time-varying magnetic signal through coil 410. Through the inverse magnetoelastic effect, the vibration of the sensor in turn generates a time varying magnetic flux, which can be measured with pick-up coil 414. A time-domain detected signal can then be converted into the frequency domain by performing a Fast Fourier Transform (FFT) to determine the resonant frequency. The resonant frequency of the transiently excited sensor can also be determined by counting the zero crossings of the sensor response for a given time period. Alternatively, the magnetoelastic sensors can be interrogated in the frequency domain by sweeping the frequency and recording the measured amplitude each incremental frequency. In various embodiments, a shift in resonance frequency of the said ME pH sensitive mass sensor depends only on the mass change of the ingestible due to the dissolution of the pH sensitive surface coating 118 of FIG. 1. In one embodiment, ME transducer driver 402 may be programmed to transmit one or more sinusoidal wave train in planar coil 410 with current passing through the coil to generate the magnetic excitation field. The emitted field can be detected by pick-up coil 414 as an exponentially decaying sinusoidal (i.e., ring down) or as an alternative by the transmitting using an isolation switch. The resonant frequency of the sensor can be determined from the ring down response using two different techniques: frequency counting and FFT. Using the FFT algorithm, the time-domain response of the sensor is converted into frequency-domain, and the resonant frequency is determined by finding the peak of the frequency domain spectrum. In an alternative implementation, a frequency-domain system requires excitation coil 410 and a pick-up (detection) coil 414. The excitation coil 410 is excited with fixed-frequency steady state signal, and the pick-up coil 414 measures the sensor response at that frequency. The frequency of the steady state signal is gradually increased, maintaining steady state operation, and the sensor response at a desired frequency range is measured with predetermined parameter programmed in logic module 404 or logic module 420. The resonant frequency of the sensor is determined by finding the frequency where the amplitude of the sensor is greatest. In various embodiments, the pH sensor is calibrated with varying thickness of layer 118 of FIG. 1 through in vitro experimentations. In a preferred embodiment, the ME pH sensor is fabricated to operate linearly in the pH range of 1 to 9 with a non-limiting change in resonant frequency of 0.2% per pH. In various embodiments, layer 118 of FIG. 1 having nonlimiting thickness between 1 to 5 pm. The ME pH sensitive mass sensor enables pH-dependent GI tract and or targeted ileocolonic drug delivery.

Aspects of the present disclosure may include methods for targeted or localized ultrasound- mediated drug delivery within the GI tract using an ME transducer, ME pH sensitive mass sensor, a pH sensitive coated ingestible integrated capsule drug delivery system. In various embodiments, one method for targeted drug delivery may comprise determining a mass change of the drug delivery device or a portion of the device caused by a dissolution of a pH sensitive polymer encapsulating at least a portion or the whole/total of the surface of the device or a pH sensitive layer of a coating at least a portion or whole/total of the surface of the device. In various embodiments, another method may comprise the synchronous or asynchronous, continuous, or intermittent monitoring and detection of a shift in the electro-mechanical resonance of an ME sensor due a change in the mass or whole or portion of the drug delivery device. Upon detection of a specific change in device whole or portion mass, the ME transducer is actuated to produce one or more timed ultrasonic vibration to release at least one drug from the drug carrier/reservoir and subsequent active transport of the drug to, into, or within a GI cell, membrane, or tissue. In another aspect, yet another method may comprise administering a therapeutic agent to a GI tissue of a subject by transporting the ingestible capsule to at least one specific location of the GI and the payload containing an encapsulated or non-encapsulated therapeutic agent is activated by an ME transducer within the capsule for control -released, pulsatile, non-pulsatile, intermittent, digital, or continuous local or targeted delivery of said agent from the payload or reservoir into GI tissue of the subject. In another aspect, said drug reservoir may be configured to releasably retain at least one encapsulated therapeutic agent. In various embodiments, the ME ultrasound transducer may be positioned to transduce ultrasound waves in a particular direction relative to the reservoir of the ingestible capsule. The ME ultrasound transducer may be positioned to transduce ultrasound waves toward the reservoir. The ME ultrasound transducer is positioned to transduce ultrasound waves away from the reservoir. The ME ultrasound transducer is positioned to produce omnidirectional ultrasound waves through the drug carrier or reservoir layer 116 of FIG. 1. The reservoir is configured to releasably retain a liquid comprising a therapeutic agent or encapsulated therapeutic agent. The resulting methods enable the implementation of a miniature low power high-volume efficient integrated ingestible capsule pH-dependent GI tract and or targeted ileocolonic drug delivery system.

Referring now to FIG. 5 a flow chart 500 of a method using ME mass sensing for targeted drug delivery using a low power high-volume efficient ingestible capsule is shown, according to various embodiments. In one embodiment, the method comprises the use of said ME pH sensor to determine a specific location with the GI for activating said ME ultrasound transducer to release and subsequently transport at least one drug to the GI tissue. In various embodiments, a patient swallows a capsule (Step 502), and the logic module 402 or 420 of FIG. 4 is programmed to monitor the resonant frequency of the ME pH sensitive mass sensor continuously or intermittently for changes in pH associated with various GI location, including but not limited to ingestion, pylorus, ileocecal valve, stomach, colon, or rectum. In a preferred embodiment, the ME pH sensor is calibrated in conjunction with layer 118 of FIG. 1 to detect the pH corresponding to the stomach, duodenum jejunum, ileum, cecum, colon, and rectum. In a step 504, the identification that capsule is ingested for compliance is monitored by a change ME sensor resonance frequency calibrated to corresponding to a dissolution rate of layer 118 of FIG. 1. In a step 506, said logic module continues to monitor the change in GI pH during the transport through the GI tract using the up-shift in the resonance frequency of the ME sensor. In a step 508, low frequency ultrasound is activated to release drug from the payload upon the dissolution of layer 118, a chosen pH level. In a preferred embodiment, for ileocolonic drug delivery, low frequency ultrasound is activated at a predetermined intensity to release drug from the payload upon the total dissolution of layer 118 at a pH greater than 7.2. In a final step 510, low frequency low ultrasound is activated at another predetermined intensity to transport released drug toward in, into, or within a GI tissue.

In another aspect, the present disclosure provides systems and methods of administering a therapeutic agent to a GI tissue of a subject by orally administering to a subject a low power high- volume efficient ingestible integrated capsule comprising a drug carrier/reservoir and a drug payload. In various embodiments, the drug payload may contain a therapeutic agent that is encapsulated in at least one pH, thermal, electric, magnetic, electromagnetic wave, catalytic, piezo-catalytic, or ultrasound-responsive polymeric carrier, including but not limited to, microbubble, nanobubble, nanodroplet, nano emulsion, nanofiber, vesicle, micelle, or hydrogel sphere or coating. In various embodiments, the ingestible capsule may comprise a coating, one or more pH sensitive coating layer or scaffold on at least one internal or external surface, said coating or scaffold contains at least one therapeutic agent encapsulated in at least one pH, thermal, electric, magnetic, electromagnetic wave, catalytic, piezo-catalytic, or ultrasound-responsive polymeric carrier. In various embodiments, the ingestible capsule may incorporate a drug carrier/reservoir containing an iron oxide particle-based biocompatible gel or microporous gel with a controlled architecture that can release its payload, containing said encapsulated or non-encapsulated therapeutic agent, when exposed to AC magnetic field or ultrasound vibration produced by at least one ME transducer of the present disclosure. In various embodiments, the said biocompatible gel or microporous gel containing at least one therapeutic agent may be deposited within one or more defined drug carrier/reservoir using, including but not limited to, at least one additive manufacturing technique. In various embodiments, said therapeutic agent may be deposited using a drop-on-demand additive manufacturing system and inkjet printing, where said drug deposited on layer 116 and enteric coating layer 118 of FIG. 1 are additively assembled in a dropwise, layer-by-layer process to achieve micron scale, multilayer structures. First, Eudragit FS 30 D is deposited onto the surface of layer 114 of FIG. 1 and evaporated utilizing the “coffee-ring” drying effect to form concave device bodies. Next, solutions of, for example RNA or a peptide (e.g., insulin) are deposited resulting in a drug payload. Finally, the drug payload is encapsulated in one or capping layers of enteric polymer, enabling controlled and delayed release of APIs, tunable to the desired pharmacokinetic profile using the ME low frequency ultrasound transducer in combination with the ME pH sensitive mass sensor of the present disclosure. This manufacturing strategy also enables on- the-fly tuning of device parameters, including device size, the mass of API and capping material deposited, and the capping material using the ME mass sensor. In an exemplary non-limiting fabrication of layer 118 of FIG. 1, Eudragit L 100 and Eudragit S 100 may be dissolved in ethyl alcohol at 1% (w/v) with 0.1% (w/v) TEC and filtered through 0.22 pm PVDF syringe filters. The depositing may be Microdevice capping was performed using the sciFLEXARRAYER S3 automated piezo driven, non-contact dispensing system. The printer may be primed with ethyl alcohol prior to printing, and the stage temperature was set to 22°C. The system can be aligned to using on or more fiduciaries. Various parameters may be optimized to obtain drops with volumes of approximately 200 to 300 pL. The printing process may be performed in multiple cycles to allow the solvent to completely evaporate between each cycle. Eudragit represents the ideal candidate for printing layer 118 of FIG. 1 due to its resistance to degradation in pH conditions < 7.0, thereby producing a carrier that will persist in the stomach for oral delivery to various specific locations of the GI, for example the stomach or the colon.

An object of the present disclosure is the encapsulation of therapeutic agents with a liquid, mixture, scaffold, or responsive polymer for incorporation into layer 116 of FIG. 1 of ingestible capsule. In various embodiments, the therapeutic agent is encapsulated in at least one pH, thermal, electric, magnetic, electromagnetic wave, catalytic, piezo-catalytic, or ultrasound-responsive polymeric carrier, including but not limited to, microbubble, nanobubble, nanodroplet, nano emulsion, nanofiber, vesicle, micelle, or hydrogel sphere or coating. In various embodiments, microbubble, nanobubble, nanodroplet, nano emulsion, nanofiber, vesicle, micelle, or hydrogel sphere or coating of the present disclosure may be produced from, but not limited to, poly(lactic acid), poly(allylamine hydrochloride), perfluorocarbon, polyvinyl alcohol, poly(lactic-co-glycolic acid, perfluoroctanol-poly(lactic acid). In various embodiments, pH or ultrasound-responsive polymer may comprise a scaffold, gel, or vesicle produce from, but not limited to, self-assembled from a poly(ethylene oxide)- block-poly[2-(diethylamino)ethyl methacrylate-stat-2-tetrahydro- furanyloxy) ethyl methacrylate] [PEO-b-P(DEA-stat- TMA)] block copolymers, polyethylene glycol) (PEG) crosslinked glycol chitosan (GC), Pluronic copolymers, poly(N ,N-diethyl acrylamide) (pNNDEA), or the like. In various embodiments, polymers for nucleic acid delivery includes, but not limited to, PS, poly(lactic acid) (PLA), poly(lactic-co-glycolic acid) (PLGA), and polyplexes of cationic polymers, polyplexes of reporter gene DNA and polyethyleneimine (PEI), poly(l-lysine)/DNA (PLL/DNA), the like, or combination thereof. Administering a therapeutic agent

Methods of administration

The invention provides methods of administering a therapeutic agent to gastrointestinal tissue of a subject using the systems and devices described above. The methods include delivering ultrasound energy to a liquid at a frequency that produces bubbles within the liquid and causes ultrasound energy, intensity, or cavitation of the bubbles. Gentle implosion of the bubbles produces shock waves that permeabilize cells and propel the agent from the liquid into the tissue. The use of ultrasound to cause cavitation to deliver agents to tissue is described in, for example, Schoellhammer, C. M., Schroeder, A., Maa, R., Lauwers, G. Y., Swiston, A., Zervas, M., et al. (2015). Ultrasound-mediated gastrointestinal drug delivery. Science Translational Medicine, 7(310), 310ral68-310ral68, doi: 10.1126/scitranslmed.aaa5937; Schoellhammer, C. M & Traverse, G., Low-frequency ultrasound for drug delivery in the gastrointestinal tract. Expert Opinion on Drug Delivery, 2016, doi: 10.1517/17425247.2016.1171841; Schoellhammer C. M., et al., Ultrasound-mediated delivery of RNA to colonic mucosa of live mice, Gastroenterology, 2017, doi: 10.1053/j.gastro.2017.01.002; and U.S. Publication Nos. 2014/0228715 and 2018/0055991, the contents of each of which are incorporated herein by reference.

In methods of the invention, the ultrasound signal may have a defined frequency. The ultrasound signal may have a frequency of from about 10 kHz to about 10 MHz, from about 10 kHz to about 1 MHz, from about 10 kHz to about 100 kHz, from about 20 kHz to about 80 kHz, from about 20 kHz to about 60 kHz, or from about 30 kHz to about 50 kHz. The ultrasound signal may have a frequency of less than 100 kHz, less than 80 kHz, less than 60 kHz, or less than 50 kHz. The ultrasound signal may have a frequency of about 20 kHz, about 25 kHz, about 30 kHz, about 35 kHz, about 40 kHz, about 45 kHz, about 50 kHz, about 55 kHz, or about 60 kHz.

In methods of the invention, the ultrasound signal may have a defined intensity. For example, and without limitation, the ultrasound signal may have an intensity of from about 0.001 W/cm ² to about 0.01 W/cm ² , from about 0.024 W/cm ² to about 0.04 W/cm ² , from about 0.014 W/cm ² to about 0.10 W/cm ² , from about 0.10 W/cm ² to about 0.5 W/cm ² , from about 0.5 W/cm ² to about .7500 W/cm ² , or from about 0.75 W/cm ² to about 1 W/cm ² .

In some embodiments, the ultrasound energy may be delivered as a pulse, i.e., it may be delivered over a brief, finite period to minimize damage to the agent being delivered by the ultrasound energy. For example, and without limitation, the pulse may be less than 20 minutes, less than 10 minutes, less than 5 minutes, or less than 10 minutes. For example, and without limitation, the pulse may be from about 10 seconds to about 3 minutes. The pulse may be about 10 minutes, about 5 minutes, about 3 minutes, about 3 minutes, about 1 minute, about 30 seconds, about 20 seconds, or about 10 seconds.

The parameters of the ultrasound pulse, such as the frequency and/or duration, may be selected so that damage to the agent is limited to a certain fraction or percentage of the agent. For example, and without limitation, the ultrasound energy may result in breakdown of less than about 95% of the agent, less than about 90% of the agent, less than about 80% of the agent, less than about 70% of the agent, less than about 60% of the agent, less than about 50% of the agent, less than about 40% of the agent, less than about 25% of the agent, or less than about 10% of the agent.

The parameters of the ultrasound pulse, such as the frequency and/or duration, may be selected so that at least a minimum amount of the agent is transferred to the tissue. For example, and without limitation, the ultrasound energy may result in transfer of at least 1% of the agent, at least 2% of the agent, at least 5% of the agent, at least 10% of the agent, at least 20% of the agent, at least 30% of the agent, or at least 40% of the agent.

The methods may be used to deliver a therapeutic agent to a specific tissue in the GI tract. For example, the tissue may be buccal tissue, gingival tissue, labial tissue, esophageal tissue, gastric tissue, intestinal tissue, colorectal tissue, or anal tissue. The therapeutic agent may be targeted to a particular tissue in the GI tract. For example, the therapeutic agent may be targeted to the stomach, small intestine, large intestine (colon), rectum, or at a duct that enters the GI tract, such as a pancreatic duct or a common bile duct.

The methods may include administering an ingestible capsule to the subject. The ingestible capsule may be administered orally or rectally. The ingestible capsule may be administered via a duct that enters the GI tract.

The methods may include positioning the ingestible capsule within the subjects GI tract. For example, the ingestible capsule may be positioned in proximity to an affected region of the GI tract, such as an ulcer or inflamed region. The ingestible capsule may be positioned by applying a magnetic field to a portion of the subject’s GI tract from a device outside the subject’s body. The magnetic field may be applied using the transmitter. Alternatively, or additionally, the magnetic field may be applied from a magnetic device that is separate from the transmitter. Therapeutic agents

The therapeutic agent may be any agent that provides a therapeutic benefit. For example and without limitation, suitable agents include alpha-hydroxy formulations, ace inhibiting agents, analgesics, anesthetic agents, anthelmintics, anti-arrhythmic agents, antithrombotic agents, antiallergic agents, anti -angiogenic agents, antibacterial agents, antibiotic agents, anticoagulant agents, anticancer agents, antidiabetic agents, anti-emetics, antifungal agents, antigens, antihypertension agents, antihypotensive agents, antiinflammatory agents, antimycotic agents, antimigraine agents, anti-obesity agents, antiparkinson agents, antirheumatic agents, antithrombins, antiviral agents, antidepressants, antiepileptics, antihistamines, antimuscarinic agents, antimycobacterial agents, antineoplastic agents, antithyroid agents, anxiolytics, asthma therapies, astringents, beta blocking agents, blood products and substitutes, bronchospamolytic agents, calcium antagonists, cardiovascular agents, cardiac glycosidic agents, carotenoids, cephalosporins, chronic bronchitis therapies, chronic obstructive pulmonary disease therapies, contraceptive agents, corticosteroids, cytostatic agents, cystic-fibrosis therapies, cardiac inotropic agents, contrast media, cough suppressants, diagnostic agents, diuretic agents, dopaminergics, elastase inhibitors, emphysema therapies, enkephalins, fibrinolytic agents, growth hormones, hemostatics, immunological agents, im- munosupressants, immunotherapeutic agents, insulins, interferons, lactation inhibiting agents, lipid-lowering agents, lymphokines, muscle relaxants, neurologic agents, NSAIDS, nutraceuticals, oncology therapies, organ-transplant rejection therapies, parasympathomimetics, parathyroid calcitonin and biphosphonates, prostacyclins, prostaglandins, psycho-pharmaceutical agents, protease inhibitors, magnetic resonance diagnostic imaging agents, radio-pharmaceuticals, reproductive control hormones, respiratory distress syndrome therapies, sedative agents, sex hormones, somatostatins, steroid hormonal agents, stimulants and anoretics, sympathomimetics, thyroid agents, vasodilating agents, vitamins, and xanthines. A more complex list of chemicals and drugs that can be used as agents in embodiments of the invention is provided in the Merck Index: An Encyclopedia of Chemicals, Drugs, and Biologicals Fifteenth Edition, Maryadele J O'Neil, ed., RSC Publishing, 2015, ISBN-13: 978-1849736701, ISBN-10 1849736707, the contents of which are incorporated herein by reference.

Therapeutic agents may be of any chemical form. For example, agents may be biological therapeutics, such as nucleic acids, proteins, peptides, polypeptides, antibodies, or other macromolecules. Nucleic acids include RNA, DNA, RNADNA hybrids, and nucleic acid derivatives that include non-naturally-occurring nucleotides, modified nucleotides, non-naturally-occurring chemical linkages, and the like. Examples of nucleic acid derivatives and modified nucleotides are described in, for example, International Publication WO 2018/118587, the contents of which are incorporated herein by reference. Nucleic acids may be polypeptide-encoding nucleic acids, such as mRNAs and cDNAs. Nucleic acids may interfere with gene expression. Examples of interfering RNAs (RNAi) include siRNAs and miRNAs. RNAi is known in the art and described in, for example, Kim and Rossi, Biotechniques. 2008 Apr; 44(5): 613-616, doi: 10.2144/000112792; and Wilson and Doudna, Molecular Mechanisms of RNA Interference, Annual Review of Biophysics 2013 42: 1, 217-239, the contents of each of which are incorporated herein by reference. Agents may be organic molecules of non-biological origin. Such drugs are often called small-molecule drugs because they typically have a molecular weight of less than 2000 Daltons, although they may be larger. Agents may be combinations or complexes of one or more biological macromolecules and/or one or more small molecules. For example, and without limitation, agents may be nucleic acid complexes, protein complexes, protein-nucleic acid complexes, and the like. Thus, the agent may exist in a multimeric or polymeric form, including homocomplexes and heterocomplexes.

An advantage of ultrasound-based delivery of therapeutic agents is the capacity to deliver large molecules, e.g., molecules having a molecular weight greater than 1000 Da. Thus, the therapeutic agent may have a minimum size. For example, and without limitation, the antigen may have a molecular weight of > 100 Da, > 200 Da, > 500 Da, > 1000 Da, > 2000 Da, > 5000 Da, > 10,000 Da, > 20,000 Da, > 50,000 Da, or > 100,000 Da.

The therapeutic agent may be provided in a liquid that promotes delivery of the therapeutic agent using the devices or systems provided herein. For example, the liquid may facilitate ultrasound-induced cavitation, iontophoresis, sonoporation, magneto-sonoporation, or electroporation. The liquid may be aqueous. The liquid may contain ions. The liquid may be an aqueous solution that contains one or more salts. The liquid may contain a buffer.

The therapeutic agent may be formulated. Formulations commonly used for delivery of biologic and small -molecule agents include drug crystals, gold particles, iron oxide particles, lipid- like particles, liposomes, micelles, microparticles, nanoparticles, polymeric particles, vesicles, viral capsids, viral particles, and complexes with other macromolecules that are not essential for the biological or biochemical function of the agent. Alternatively, the therapeutic agent may be unformulated, i.e., it may be provided in a biologically active format that does not contain other molecules that interact with the agent solely to facilitate delivery of the agent. Thus, the agent may be provided in a non-encapsulated form or in a form that is not complexed with other molecules unrelated to the function of the agent.

The agent may be a component of a gene editing system, such as a meganuclease, zinc finger nuclease (ZFN), a transcription activator-like effector-based nuclease (TALEN), or the clustered, regularly-interspersed palindromic repeat (CRISPR) system.

Meganucleases are endodeoxyribonucleases that recognize double-stranded DNA sequences of 12-40 base pairs. They can be engineered to bind to different recognition sequences to create customized nucleases that target particular sequences. Meganucleases exist in archaebacte- rial, bacteria, phages, fungi, algae, and plants, and meganucleases from any source may be used. Engineering meganucleases to recognize specific sequences is known in the art and described in, for example, Stoddard, Barry L. (2006) "Homing endonuclease structure and function" Quarterly Reviews of Biophysics 38 (1): 49-95 doi: 10.1017/S0033583505004063, PMID 16336743; Grizot, S.; Epinat, J. C.; Thomas, S.; Duclert, A.; Rolland, S.; Paques, F.; Duchateau, P. (2009) "Generation of redesigned homing endonucleases comprising DNA-binding domains derived from two different scaffolds" Nucleic Acids Research 38 (6): 2006-18, doi: 10.1093/nar/gkpl l71. PMC 2847234, PMID 20026587; Epinat, Jean-Charles; Arnould, Sylvain; Chames, Patrick; Rochaix, Pascal; Desfontaines, Dominique; Puzin, Clemence; Patin, Amelie; Zanghellini, Alexandre; Paques, Frederic (2003-06-01) "A novel engineered meganuclease induces homologous recombination in yeast and mammalian cells" Nucleic Acids Research 31 (11): 2952-2962; and Seligman, L. M.; Chisholm, KM; Chevalier, BS; Chadsey, MS; Edwards, ST; Savage, JH; Veillet, AL (2002) "Mutations altering the cleavage specificity of a homing endonuclease" Nucleic Acids Research 30 (17): 3870-9, doi:10.1093/nar/gkf495. PMC 137417, PMID 12202772, the contents of each of which are incorporated herein by reference.

ZFNs are artificial restriction enzymes that have a zinc finger DNA-binding domain fused to a DNA-cleavage domain. ZFNs can also be engineered to target specific DNA sequences. The design and use of ZFNs is known in the art and described in, for example, Carroll, D (2011) "Genome engineering with zinc-finger nucleases" Genetics Society of America 188 (4): 773-782, doi: 10.1534/genetics.l l l.131433. PMC 3176093, PMID 21828278; Cathomen T, Joung JK (July 2008) "Zinc-finger nucleases: the next generation emerges" Mol. Ther. 16 (7): 1200-7, doi: 10.1038/mt.2008.114, PMID 18545224; Miller, J. C.; Holmes, M. C.; Wang, J.; Guschin, D. Y.; Lee, Y. L.; Rupniewski, I.; Beausejour, C. M.; Waite, A. J.; Wang, N. S.; Kim, K. A.; Gregory, P. D.; Pabo, C. 0.; Rebar, E. J. (2007) "An improved zinc-finger nuclease architecture for highly specific genome editing" Nature Biotechnology, 25 (7): 778-785, doi: 10.1038/nbtl319, PMID 17603475, the contents of each of which are incorporated herein by reference.

TALENs are artificial restriction enzymes that have a TAL effector DNA-binding domain fused to a DNA cleavage domain. TALENs can also be engineered to target specific DNA sequences. The design and use of TALENs is known in the art and described in, for example, Boch J (February 2011) "TALEs of genome targeting" Nature Biotechnology 29 (2): 135-6, doi:10.1038/nbt.1767. PMID 21301438; Juillerat A, Pessereau C, Dubois G, Guyot V, Marechai A, Valton J, Daboussi F, Poirot L, Duclert A, Duchateau P (January 2015) "Optimized tuning of TALEN specificity using non-conventional RVDs" Scientific Reports, 5: 8150, doi: 10.1038/srep08150. PMC 4311247, PMID 25632877; and Mahfouz MM, Li L, Shamimuz- zaman M, Wibowo A, Fang X, Zhu JK (February 2011) "De novo-engineered transcription activator-like effector (TALE) hybrid nuclease with novel DNA binding specificity creates doublestrand breaks" Proceedings of the National Academy of Sciences of the United States of America, 108 (6): 2623-8, Bibcode:2011PNAS,1O8.2623M, doi: 10.1073/pnas.1019533108, PMC 3038751, PMID 21262818, the contents of each of which are incorporated herein by reference.

The CRISPR system is a prokaryotic immune system that provides acquired immunity against foreign genetic elements, such as plasmids and phages. CRISPR systems include one or more CRISPR-associated (Cas) proteins that cleave DNA at clustered, regularly-interspersed palindromic repeat (CRISPR) sequences. Cas proteins include helicase and exonuclease activities, and these activities may be on the same polypeptide or on separate polypeptides. Cas proteins are directed to CRISPR sequences by RNA molecules. A CRISPR RNA (crRNA) binds to a complementary sequence in the target DNA to be cleaved. A transactivating crRNA (tracrRNA) binds to both the Cas protein and the crRNA to draw the Cas protein to the target DNA sequence. Not all CRISPR systems require tracrRNA. In nature crRNA and tracrRNA occur on separate RNA molecules, but they also function when contained a single RNA molecule, called a single guide RNA or guide RNA (gRNA). The one or more RNAs and one or more polypeptides assemble inside the cell to form a ribonucleoprotein (RNP). CRISPR systems are described, for example, in van der Oost, et al., CRISPR-based adaptive and heritable immunity in prokaryotes, Trends in Biochemical Sciences, 34(8):401 -407 (2014); Garrett, et al., Archaeal CRISPR-based immune systems: exchangeable functional modules, Trends in Microbiol. 19(11):549-556 (2011); Makarova, et al., Evolution and classification of the CRISPR-Cas systems, Nat. Rev. Microbiol. 9:467-477 (2011); and Sorek, et al., CRISPR-Mediated Adaptive Immune Systems in Bacteria and Archaea, Ann. Rev. Biochem. 82:237-266 (2013), the contents of each of which are incorporated herein by reference.

CRISPR-Cas systems have been placed in two classes. Class 1 systems use multiple Cas proteins to degrade nucleic acids, while class 2 systems use a single large Cas protein. Class 1 Cas proteins include CaslO, CaslOd, Cas3, Cas5, Cas8a, Cmr5, Csel, Cse2, Csfl, Csm2, Csxl l, Csyl, Csy2, and Csy3. Class 2 Cas proteins include C2cl, C2c2, C2c3, Cas4, Cas9, Cpfl, and Csn2.

CRISPR-Cas systems are powerful tools because they allow gene editing of specific nucleic acid sequences using a common protein enzyme. By designing a guide RNA complementary to a target sequence, a Cas protein can be directed to cleave that target sequence. In addition, although naturally-occurring Cas proteins have endonuclease activity, Cas proteins have been engineered to perform other functions. For example, endonuclease-deactivated mutants of Cas9 (dCas9) have been created, and such mutants can be directed to bind to target DNA sequences without cleaving them. dCas9 proteins can then be further engineered to bind transcriptional activators or inhibitors. As a result, guide sequences can be used to recruit such CRISPR complexes to specific genes to turn on or off transcription. Thus, these systems are called CRISPR activators (CRISPRa) or CRISPR inhibitors (CRISPRi). CRISPR systems can also be used to introduce sequence-specific epigenetic modifications of DNA, such acetylation or methylation. The use of modified CRISPR systems for purposes other than cleavage of target DNA are described, for example, in Dominguez, et al., Beyond editing: repurposing CRISPR-Cas9 for precision genome regulation and interrogation, Nat. Rev. Cell Biol. 17(1): 5- 15 (2016), which is incorporated herein by reference.

The agent may be any component of a CRISPR system, such as those described above. For example and without limitation, the CRISPR component may be one or more of a helicase, endonuclease, transcriptional activator, transcriptional inhibitor, DNA modifier, gRNA, crRNA, or tra- crRNA. The CRISPR component contain a nucleic acid, such as RNA or DNA, a polypeptide, or a combination, such as a RNP. The CRISPR nucleic acid may encode a functional CRISPR component. For example, the nucleic acid may be a DNA or mRNA. The CRISPR nucleic acid may itself be a functional component, such as a gRNA, crRNA, or tracrRNA.

The agent may include an element that induces expression of the CRISPR component. For example, expression of the CRISPR component may be induced by an antibiotic, such as tetracycline, or other chemical. Inducible CRISPR systems have been described, for example, in Rose, et al., Rapidly inducible Cas9 and DSB-ddPCR to probe editing kinetics, Nat. Methods, 14, pages 891-896 (2017); and Cao, et al., An easy and efficient inducible CRISPR/Cas9 platform with improved specificity for multiple gene targeting, Nucleic Acids Res. 14( 19): e 149 (2016), the contents of which are incorporated herein by reference. The inducible element may be part of the CRISPR component, or it may be a separate component.

In certain embodiments of the invention, methods allow delivery of agents that promote wound healing. The agent may promote healing by any mechanism. For example and without limitation, the agent may facilitate one or more phases of the wound healing process; prevent infection, including bacterial or viral infection; or alleviate pain or sensitivity.

A variety of growth factors promote wound healing. For example and without limitation, growth factors that promote wound healing include CTGF/CCN2, EGF family members, FGF family members, G-CSF, GM-CSF, HGF, HGH, HIF, histatin, hyaluronan, IGF, IL-1, IL-4, IL-8, KGF, lactoferrin, lysophosphatidic acid, NGF, a PDGF, TGF-P, and VEGF. The EFG family includes 10 members: amphiregulin (AR), betacellulin (BTC), epigen, epiregulin (EPR), heparin- binding EGF-like growth factor (HB-EGF), neuregulin-1 (NRG1), neuregulin-2 (NRG2), neureg- ulin-3 (NRG3), neuregulin-4 (NRG4), or transforming growth factor-a (TGF-a). The FGF family includes 22 members: FGF1, FGF2 (also called basic FGF or bFGF), FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, FGF9, FGF10, FGF11, FGF12, FGF13, FGF14, FGF16, FGF17, FGF18, FGF19, FGF20, FGF21, FGF22, or FGF23. PDGF exists in three forms: PDGF AA, PDGF AB, and PDGF BB. The TGF-P family includes three forms: TGF-pi, TGF-P2, and TGF-P3.

A variety of agents that prevent infection have been used to treat wounds. For example, and without limitation, the agent may be an antimicrobial, antiviral, antibiotic, antifungal, or antiseptic. Exemplary agents include silver, iodine, chlorhexidine, hydrogen peroxide, lysozyme, peroxidase, defensins, cystatins, thrombospondin, and antibodies. Nitric oxide donors, such as glyceryl trinitrate and nitrite salts, are also useful to prevent infection and promote wound healing. Diseases, disorders, and conditions

The methods are useful to treat conditions of the GI tract of a subject. The condition may be any disease, disorder, or condition that affects the GI tract.

In some embodiments, the disorder is a disorder of the esophagus, including, but not limited to, esophagitis - (candidal), gastroesophageal reflux disease (gerd); laryngopharyngeal reflux (also known as extraesophageal reflux disease/eerd); rupture (Boerhaave syndrome, Mallory- Weiss syndrome); UES - (Zenker's diverticulum); LES - (Barrett's esophagus); esophageal motility disorder - (nutcracker esophagus, achalasia, diffuse esophageal spasm); esophageal stricture; and megaesophagus.

In some embodiments, the disorder is a disorder of the stomach, including but not limited to gastritis (e.g., atrophic, Menetrier's disease, gastroenteritis); peptic (i.e., gastric) ulcer (e.g., Cushing ulcer, Dieulafoy's lesion); dyspepsia; emesis; pyloric stenosis; achlorhydria; gastropare- sis; gastroptosis; portal hypertensive gastropathy; gastric antral vascular ectasia; gastric dumping syndrome; and human mullular fibrillation syndrome (HMFS).

In some embodiments, the disorder is a disorder of the small intestine, including but not limited to, enteritis (duodenitis, jejuni tis, ileitis); peptic (duodenal) ulcer (curling's ulcer); malabsorption: celiac; tropical sprue; blind loop syndrome; Whipple's; short bowel syndrome; steatorrhea; Milroy’s disease In some embodiments, the disorder is a disorder of the small intestine, including but not limited to, both large intestine and small intestine enterocolitis (necrotizing); inflammatory bowel disease (IBD); Crohn's disease; vascular; abdominal angina; mesenteric ischemia; angiodysplasia; bowel obstruction: ileus; intussusception; volvulus; fecal impaction; constipation; and diarrhea.

In some embodiments, the disorder is a disorder of the small intestine, including but not limited to, accessory digestive glands disease; liver hepatitis (viral hepatitis, autoimmune hepatitis, alcoholic hepatitis); cirrhosis (PBC); fatty liver (Nash); vascular (hepatic veno-occlusive disease, portal hypertension, nutmeg liver); alcoholic liver disease; liver failure (hepatic encephalopathy, acute liver failure); liver abscess (pyogenic, amoebic); hepatorenal syndrome; peliosis hepatis; hemochromatosis; and Wilson's disease.

In some embodiments, the disorder is a disorder of the pancreas, including, but not limited to, pancreas pancreatitis (acute, chronic, hereditary); pancreatic pseudocyst; and exocrine pancreatic insufficiency. In some embodiments, the disorder is a disorder of the large intestine, including but not limited to, appendicitis; colitis (pseudomembranous, ulcerative, ischemic, microscopic, collagenous, lymphocytic); functional colonic disease (IBS, intestinal pseudoobstruction/ogilvie syndrome); megacolon/toxic megacolon; diverticulitis; and diverticulosis.

In some embodiments, the disorder is a disorder of the large intestine, including but not limited to, gall bladder and bile ducts, cholecystitis; gallstones/cholecystolithiasis; cholesterolosis; Rokitansky-Aschoff sinuses; postcholecystectomy syndrome cholangitis (PSC, ascending); cho- lestasis/Mirizzi's syndrome; biliary fistula; haemobilia; and gallstones/cholelithiasis. In some embodiments, the disorder is a disorder of the common bile duct (including choledocholithiasis, biliary dyskinesia).

Other disorders which can be treated with the methods and devices included herein include acute and chronic immune and autoimmune pathologies, such as systemic lupus erythematosus (SLE), rheumatoid arthritis, thyroidosis, graft versus host disease, scleroderma, diabetes mellitus, Graves' disease, Behcet's disease; inflammatory diseases, such as chronic inflammatory pathologies and vascular inflammatory pathologies, including chronic inflammatory pathologies such as sarcoidosis, chronic inflammatory bowel disease, ulcerative colitis, and Crohn's pathology and vascular inflammatory pathologies, such as, but not limited to, disseminated intravascular coagulation, atherosclerosis, giant cell arteritis and Kawasaki's pathology; malignant pathologies involving tumors or other malignancies, such as, but not limited to leukemias (acute, chronic myelocytic, chronic lymphocytic and/or myelodysplastic syndrome); lymphomas (Hodgkin's and non-Hodg- kin's lymphomas, such as malignant lymphomas (Burkitt's lymphoma or Mycosis fungoides)); carcinomas (such as colon carcinoma) and metastases thereof; cancer-related angiogenesis; infantile hemangiomas; and infections, including, but not limited to, sepsis syndrome, cachexia, circulatory collapse and shock resulting from acute or chronic bacterial infection, acute and chronic parasitic and/or infectious diseases, bacterial, viral or fungal, such as a HIV, AIDS (including symptoms of cachexia, autoimmune disorders, AIDS dementia complex and infections).

Other disorders which can be treated with the methods and devices included herein include acute and chronic immune and autoimmune pathologies, inflammatory diseases, infections, and malignant pathologies involving, e.g., tumors or other malignancies.

The subject suffering from the GI condition may be any type of subject, such as an animal, for example, a mammal, for example, a human. Incorporation by Reference

References and citations to other documents, such as patents, patent applications, patent publications, journals, books, papers, web contents, have been made throughout this disclosure. All such documents are hereby incorporated herein by reference in their entirety for all purposes.

Equivalents

Various modifications of the invention and many further embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art from the full contents of this document, including references to the scientific and patent literature cited herein. The subject matter herein contains important information, exemplification, and guidance that can be adapted to the practice of this invention in its various embodiments and equivalents thereof.