The present invention relates generally to methods, formulations and systems for delivery of imaging and therapeutic agents to the middle or inner ear.
Clinical diagnosis and treatment of ear infections is a $4 Billion market. In particular, otitis media (OM, inflammation in the middle ear) is one of the most common illnesses among children under five years of age. It is known to affect about 90% of children worldwide. Only in the US, 2.2 million cases of OM are diagnosed annually at a cost of $4 billion. Accurate diagnosis and treatment of OM is critical as it can lead to additional complications such as speech and language development delays, brain abscesses, meningitis, and permanent hearing loss. Additional information is provided in the infographic of
There are several current standards of diagnosis and treatment for persistent OM however each has a variety of limitations.
One mode of diagnosis for detecting the presence of fluid in the middle ear by pneumatic otoscopy using a light, magnifying lenses to examine monitor the motion of the eardrum in response to a puff of air. This method is subject to erroneous interpretation by the practitioner and thus highly prone to error. However, this mode remains the current standard of practice.
Still another conventional mode includes treatment by surgical insertion of a tube into the tympanic membrane (TM) and removal the thick fluid by suction. This procedure suffers from the shortcomings of requiring general anesthesia and multiple post-operative check-ups until the tube extrudes and the tympanic membrane heals.
Still another conventional mode includes treatment by antibiotic administration. It is common to use the delivery modes of systemic intravenous injection or oral administration. However, these delivery modes suffer from non-specific biodistribution and frequently require high-dosage administration.
What is needed are improvements in the diagnosis and treatment of middle and inner ear disorders.
In general, in one embodiment, a method of detecting or treating a disorder in an ear of a patient includes delivering an ear drop containing a nanoparticle having at least one of an imaging agent, a diagnostic agent, or a therapeutic agent into the ear canal of the patient, penetrating without damaging the tympanic membrane or round window with the nanoparticle, and delivering the agent or agents to the middle ear or the inner ear of the patient for the detection or treatment of a disorder of ear of the patient.
This and other embodiments can include one or more of the following features. Multiple imaging agents, diagnostic agents, or therapeutic agents can be delivered by the same nanoparticle. The nanoparticle can be adapted for detection or treatment of otitis media. The nanoparticle can be adapted for detection or changing properties of fluid as part of a diagnosis or treatment of otitis media with effusion. The nanoparticle can be adapted for detection or eradication of infection in fluid as part of a diagnosis or treatment of acute or chronic otitis media. The imaging agent can include an inflammation targeted probe that will fluoresce further including a step of detecting a presence of a fluid and/or an indication of whether the fluid is infected. The probe can fluoresce within the visible, NIR and short-wave infrared spectrum. The nanoparticle can be a lipid-based or liposomal formulation.
In general, in one embodiment, a liposomal nanoparticle for delivery to structures of the middle or inner ear includes a vesicular structure composed of lipids arranged in a shell-like bilayer formulated for trans-tympanic membrane delivery, and at least one of an imaging agent, a diagnostic agent, or a therapeutic agent loaded onto the structure.
This and other embodiments can include one or more of the following features. The liposomal nanoparticle can further include a formulation adapted for delivery of mucolytic or anti-microbial drugs to the middle ear. The liposomal nanoparticle can further include a formulation adapted for delivery of therapeutic or protective agents to the inner ear. The liposomal nanoparticle can further include a formulation including multiple compounds loaded simultaneously on, in or within the liposomal nanoparticle. The lipids can include phospholipids. The lipids can include a hydrophilic surface and core and a hydrophobic interior of the shell like bi-layer. The at least one of the imaging agents, the diagnostic agent or the therapeutic agent can be loaded on, in or within a portion of a hydrophilic surface, a hydrophilic core, or interior or combinations thereof. The liposomal nanoparticle can be formulated as an ear drop.
The novel features of the invention are set forth with particularity in the claims that follow. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:
In one aspect of the present invention, there is provided a method and system for delivering imaging agents, therapeutic agents, or both in a single formulation to the middle or inner ear area using nanoparticles that can pass through the tympanic membrane (i.e., eardrum).
The anatomy of the human ear is provided in the views of
In the description that follows, it is to be appreciated that one can load the agents anywhere on the nanoparticle, including hydrophilic surface, hydrophilic core, or both. As such the agents may be considered broadly as loaded into the nanoparticle structure as variously described herein.
Still further, embodiments of the present invention include an ear drop based on liposomal nanoparticles that penetrate the eardrum and round window membrane (RWM) without an incision and can deliver therapeutic agents and/or contrast agents to the middle ear and inner ear. Advantageously as compared to the conventional methods detailed above, embodiments of the inventive delivery system are inexpensive, painless, and risk-free alternative to surgery and extremely convenience for patients, resulting in fewer doctor visits, reducing cost of care, and improving overall quality of life for children and their parents. In additional alternatives, there are embodiments that provide a health care practitioner with a local delivery pathway including an ability to concentrate therapeutic and imaging agents for middle and inner ear diseases. In still other alternatives, there are provided a wide range of alternative embodiments of this method, using other types of nanoparticle agents, imaging probes, drug molecules, and administrative procedures can be employed for the various uses described herein.
In one aspect, there is provided a formulated and synthesized a liposomal nanoparticle carrier. In a specific embodiment, there are included vesicular structures composed of phospholipids arranged in a shell-like bilayer with a hydrophilic surface (facing aqueous solution) and hydrophobic interior. Embodiments of this shell-like structure makes liposomes ideal carriers, enabling loading or encapsulation of drug molecules. The ability of the inventive embodiments to fuse (at nanoscale) with lipid-rich barriers in the tympanic membrane enables liposomes to push the cargo (e.g., imaging probes or drug molecules) through without damaging those barriers. Embodiments of the liposomes as described herein are biocompatible (i.e., non-toxic) and bio-degradable, in formulations safe for administration to pediatric patients. Safe as used herein refers to meeting those standards used for liposomes when first FDA-approved as nanomedicines for clinical trials in the 1990s. In still further embodiments, the inventive liposomes are adapted liposomal nano-formulations suited for trans-tympanic delivery. In this context, suited for trans-tympanic delivery refers to embodiment that can carry encapsulated/conjugated therapeutic and contrast agents through the TM alone or in combination with the RWM for therapeutic or diagnostic applications in disorders of the middle ear and inner ear or other disease states.
While desiring not to be bound by theory, it is believed that the tympanic membrane has a similar structure and penetration barriers like the skin consisting of keratin and a lipid-rich stratum corneum.
The RWM consists of three layers: an outer epithelial layer facing the middle ear, a central connective tissue layer, and an inner epithelial layer. The evidence shows the RWM permeability allows passage of a wide range of materials including antibiotics, local anesthetics, toxins, and albumin. As such, it is believed that liposomal nanoparticles suited for topical delivery (i.e., delivery through skin) may be well suited and are adapted in these various formulations and embodiment for various applications of ototopical delivery (i.e., delivery through eardrum) for diagnostic, therapeutic, imaging or other payloads as described herein, or any of those in
In still other embodiments, aspects of the present invention may be provided in a form factor allowing use as an ear drop that can be administered clinically by physicians or other health practitioners in the care of pediatric patients using a non-invasive diagnosis and treatment of their middle ear infections. In still further embodiments, there is provided a method for diagnosis and treatment of other middle/inner ear related diseases in a convenient, cost-effective, and painless fashion including additional formulations incorporated into a trans-tympanic liposomal formulation including agents developed by pharmaceutical companies or biotech companies. Additionally or optionally, the advantageous trans-tympanic formulations and techniques may can be used for combined diagnosis and therapy (i.e., using one agent to do both) as well as multifunctional diagnosis or therapy (i.e., delivering one or more probes or drugs with different properties, e.g., inserting hydrophobic molecule within the shell layers and encapsulating hydrophilic molecules at the core and/or loading them on the surface, within the same liposomal carrier). One exemplary ear related disease suited for treatment using an embodiment of the present invention includes sensorineural hearing loss (SNHL).
In one illustrative embodiment, liposomal vesicles formulation, synthesis, and characterization were fabricated as follows: First, there was a thin-film hydration followed by extrusion method to form elastic liposomal vesicles. Soybean phosphatidylcholine (PC) with purity >99% and sodium cholate were used as lipid source and as surfactant to enhance the elasticity of the liposomes respectively. The obtained vesicles were downsized by passing through an extruder system equipped to a polycarbonate filter with 100 nm pore size. The filtrated transferosomes were lyophilized in presence of sucrose to avoid any undesirable aggregation/fusion.
The vesicles size distribution was then evaluated as shown in
Cryo-TEM images described above showed uniform and unilamellar formation of vesicles with apparent size at around 120 nm and confirmed the vesicles stayed intact without any aggregation after freeze-drying and encapsulation with the fluorescent RhB-dye.
Further consideration of the various images in
These confocal images from inner ear show particle penetrations beyond the middle ear to inner ear. (
For analysis of hair cells patterns in the organ of corti, the otic capsule containing the membranous inner ear were dissected out by decapitation and opening of the skull at the midline. The whole inner ear was fixed in 4% PFA in PBS for overnight and decalcified with EDTA solution (100 mM) for three days.
We studied long-term effects of our formulation on the functionality of the mice ears by testing their hearing ability via auditory brainstem response (ABR). The results showed that hearing sensitivities remained largely unchanged demonstrating that vesicles neither disturb the function of the inner ear and nor causes hearing impairment in mice.
We performed an MTT assay to evaluate the safety of the designed formulations for subsequent clinical trials. Vocal fold fibroblasts (VFF) cells were exposed to the vesicles at different concentrations and the viability of the cells was quantified by measuring using a microplate reader. MTT cytotoxicity studies on vocal cells after 24 h of incubation with vesicles. The results indicate that vesicles did not affect the proliferation of the cells, hence are not toxic.
Embodiments of the above may be employed in a wide variety of methods of detecting or treating a disorder in an ear of a patient. For example, in the exemplary method 100 in
In still other aspects or alternatives, the liposomal nanoparticle is adapted for detection of inflammatory fluid in the middle ear and an indication of infection in the detected fluids as part of a diagnosis of otitis media. In still other aspects, the imaging agent comprises an inflammation targeted probe that fluoresce comprising a step of detecting through the tympanic layer a presence of a bacterial fluid and an indication of whether the fluid is infected based on the response of the targeted probe that can fluoresce within the visible, NIR and short-wave infrared spectrum.
In other alternatives the liposomes can deliver mucolytic medications to the middle ear to decrease the viscosity of the middle ear fluid. Additionally or optionally, in some embodiments the nanoparticles may deliver mucolytic drugs to the middle ear fluid, which can soften the fluid (e.g., reduce its viscosity and stickiness) thereby facilitating its discharge from the nose of the patient.
In still other alternative embodiments, formulations of the inventive nanoparticle may be adapted for combination therapy to enhance the efficacy of the treatment, or still further as a multi-modal solution that can be used for imaging and therapy enabled by the delivery of a single product into the ear canal.
In still further alternatives, an embodiment of the nanoparticles may deliver antibiotics or combinations of antibiotics and other anti-microbial drugs to eradicate bacterial infection in the middle ear. In still further embodiments, there is provided a formulation of the nanoparticle adapted for treatment, removal or elimination of bacteria based or other undesired biofilms in the middle ear. Additionally, or optionally, the nanoparticle formulation may be formulated to specifically target those bacteria or biofilms that are hard to penetrate or treat with antibiotics. In still further alternatives, the liposomal nanoparticle comprises lipids adapted for penetration of biofilms within the ear.
While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.
When a feature or element is herein referred to as being “on” another feature or element, it can be directly on the other feature or element or intervening features and/or elements may also be present. In contrast, when a feature or element is referred to as being “directly on” another feature or element, there are no intervening features or elements present. It will also be understood that, when a feature or element is referred to as being “connected”, “attached” or “coupled” to another feature or element, it can be directly connected, attached or coupled to the other feature or element or intervening features or elements may be present. In contrast, when a feature or element is referred to as being “directly connected”, “directly attached” or “directly coupled” to another feature or element, there are no intervening features or elements present. Although described or shown with respect to one embodiment, the features and elements so described or shown can apply to other embodiments. It will also be appreciated by those of skill in the art that references to a structure or feature that is disposed “adjacent” another feature may have portions that overlap or underlie the adjacent feature.
Terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. For example, as used herein, the singular forms “a”, “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “comprises” and/or “comprising,” when used in this specification, specify the presence of stated features, steps, operations, elements, and/or components, but do not preclude the presence or addition of one or more other features, steps, operations, elements, components, and/or groups thereof. As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items and may be abbreviated as “/”.
Spatially relative terms, such as “under”, “below”, “lower”, “over”, “upper” and the like, may be used herein for ease of description to describe one element or feature’s relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is inverted, elements described as “under” or “beneath” other elements or features would then be oriented “over” the other elements or features. Thus, the exemplary term “under” can encompass both an orientation of over and under. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly. Similarly, the terms “upwardly”, “downwardly”, “vertical”, “horizontal” and the like are used herein for the purpose of explanation only unless specifically indicated otherwise.
Although the terms “first” and “second” may be used herein to describe various features/elements (including steps), these features/elements should not be limited by these terms, unless the context indicates otherwise. These terms may be used to distinguish one feature/element from another feature/element. Thus, a first feature/element discussed below could be termed a second feature/element, and similarly, a second feature/element discussed below could be termed a first feature/element without departing from the teachings of the present invention.
Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise”, and variations such as “comprises” and “comprising” means various components can be co-jointly employed in the methods and articles (e.g., compositions and apparatuses including device and methods). For example, the term “comprising” will be understood to imply the inclusion of any stated elements or steps but not the exclusion of any other elements or steps.
As used herein in the specification and claims, including as used in the examples and unless otherwise expressly specified, all numbers may be read as if prefaced by the word “about” or “approximately,” even if the term does not expressly appear. The phrase “about” or “approximately” may be used when describing magnitude and/or position to indicate that the value and/or position described is within a reasonable expected range of values and/or positions. For example, a numeric value may have a value that is +/- 0.1% of the stated value (or range of values), +/- 1% of the stated value (or range of values), +/- 2% of the stated value (or range of values), +/- 5% of the stated value (or range of values), +/- 10% of the stated value (or range of values), etc. Any numerical values given herein should also be understood to include about or approximately that value, unless the context indicates otherwise. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Any numerical range recited herein is intended to include all sub-ranges subsumed therein. It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value” and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “X” is disclosed the “less than or equal to X” as well as “greater than or equal to X” (e.g., where X is a numerical value) is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point “15” are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.
Although various illustrative embodiments are described above, any of a number of changes may be made to various embodiments without departing from the scope of the invention as described by the claims. For example, the order in which various described method steps are performed may often be changed in alternative embodiments, and in other alternative embodiments one or more method steps may be skipped altogether. Optional features of various device and system embodiments may be included in some embodiments and not in others. Therefore, the foregoing description is provided primarily for exemplary purposes and should not be interpreted to limit the scope of the invention as it is set forth in the claims.
The examples and illustrations included herein show, by way of illustration and not of limitation, specific embodiments in which the subject matter may be practiced. As mentioned, other embodiments may be utilized and derived there from, such that structural and logical substitutions and changes may be made without departing from the scope of this disclosure. Such embodiments of the inventive subject matter may be referred to herein individually or collectively by the term “invention” merely for convenience and without intending to voluntarily limit the scope of this application to any single invention or inventive concept, if more than one is, in fact, disclosed. Thus, although specific embodiments have been illustrated and described herein, any arrangement calculated to achieve the same purpose may be substituted for the specific embodiments shown. This disclosure is intended to cover any and all adaptations or variations of various embodiments. Combinations of the above embodiments, and other embodiments not specifically described herein, will be apparent to those of skill in the art upon reviewing the above description.
This application is a continuation of co-pending International Application No. PCT/US2021/ 044155, filed Oct. 15, 2021, which claims the benefit of U.S. Provisional Pat. Application No. 63/092,497, filed Oct. 15, 2020, titled “METHOD AND SYSTEM FOR OTOTOPICAL DELIVERY USING NANOPARTICLES FOR EAR INFECTION DIAGNOSIS AND TREATMENT,” the entire disclosures of which are herein incorporated by reference. All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
This invention was made with Government support under contract TR003142 awarded by the National Institutes of Health. The Government has certain rights in the invention.
Number | Date | Country | |
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63092497 | Oct 2020 | US |
Number | Date | Country | |
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Parent | PCT/US2021/044155 | Aug 2021 | WO |
Child | 18132290 | US |