Methods for administering therapeutic agents to the subretinal space

Abstract

The present disclosure relates to a method of administering a therapeutic agent to the subretinal space of a subject's eye, the method comprising the steps of creating a localized retinal detachment in the subretinal space of the subject's eye, injecting a composition comprising a magnetized therapeutic agent and a volume of a pharma- ceutically acceptable carrier, and applying a magnetic force to the eye, the magnetic force adhering the magnetized therapeutic agent to the subretinal space of the eye. Additionally, the method comprises removing at least 50% of the volume of the pharma- ceutically acceptable carrier from the eye while the magnetic force is applied to the eye.

Description

Related Applications

[0001] [Related Applications]

[0001] This application claims priority to and benefit of U.S. Provisional Patent Application No. 63 / 311,718, filed February 18, 2022, by Emmetrope Ophthalmics LLC, entitled “Methods of Administrating Therapeutic Agent to the Subretinal Space,” the entire disclosure of which is hereby incorporated by reference into this specification. [Technical field]

[0002] This document relates to methods of administering therapeutic agents to the subretinal space of the eye in a subject and methods of treating retinal disorders in a patient. [Background technology]

[0003]

[0003] Cellular therapies for ocular regeneration, such as cell injections for endothelial cell dysfunction and stem cell transplantation for retinal neuroprotection, have been investigated, but these procedures are technically challenging, have variable success rates, and are not yet commercially available for many types of ocular diseases, including: diseases of the cornea (including but not limited to endothelial dystrophies), diseases of the retinal ganglion cells and optic nerve (including but not limited to glaucoma, ischemic optic neuropathy, and other optic neuropathies), and diseases of the retinal photoreceptors and retinal pigment epithelium (including but not limited to Leber's congenital amaurosis, retinitis pigmentosa, and age-related macular degeneration).

[0004]

[0004] Unlike cell transplantation therapy in other organs, simply injecting therapeutic cells or other therapeutic agents into the eye is generally ineffective because they remain localized and do not attach to or integrate into the patient's tissue. For example, when healthy corneal endothelial cells are injected into the anterior chamber of the eye, they do not integrate very efficiently in preclinical models of corneal endothelial dysfunction (e.g., Mimura et al., Invest. Ophthalmol. Vis. Sci. 2005, 46(10): 3637-44). Similarly, healthy retinal ganglion cells do not integrate into the correct retinal layers when simply injected into the cavity of the eye. For the eye, most current cell therapy techniques lack techniques for controlling cell localization in vivo. Stem cell transplant clinical trials for retinitis pigmentosa use subretinal injections of hematopoietic stem cells, but have no mechanism to control the localization of stem cells to the subretinal space to prevent them from floating or migrating in the ocular fluid. As another example, corneal endothelial cells injected into the anterior chamber of the eye simply fall off the cornea by gravity and do not attach properly unless they are co-injected with molecules that promote cell adhesion (Kinoshita et al., N Engl J Med. 2018, 378(11): 995-1003). Thus, new methods to target cells to specific tissues for therapeutic purposes are long needed. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] U.S. Provisional Patent Application No. 63 / 311,718 [Non-patent literature]

[0006] [Non-Patent Document 1] Mimura et al., Invest. Ophthalmology and Visual Science, 2005, 46(10): 3637-44. [Non-Patent Document 2] Kinoshita et al., N Engl J Med. 2018, 378(11): 995-1003 [Non-Patent Document 3] Remington: The Science and Practice of Pharmacy, 21st Ed. (2005) [Non-Patent Document 4] Neuwell et al., Neurosurgery. 1994, 34: 777-784 [Non-Patent Document 5] Schutt et al., Hybridoma. 1997, 16: 109-117 [Non-Patent Document 6] Lubbe et al., J Surg Res. 2001, 95: 200-206 [Non-Patent Document 7] Miltenyi et al., Cytometry. 1990, 11:231-238 [Non-Patent Document 8] Schroder et al., J Immunol Methods. 1986, 93: 45-53 [Non-Patent Document 9] Douglas et al., Crit Rev Ther Drug Carrier Syst. 1987, 3: 233-261 [Non-Patent Document 10] Sestier et al., Electrophoresis. 1998, 19: 1220-1226 [Non-Patent Document 11] Perrin et al., J Immunol Methods. 1999, 224:77-87 [Non-Patent Document 12] McCloskey et al., Cytometry. 2000, 40: 307-315 [Non-Patent Document 13] Tibbe et al., Cytometry. 2001, 43: 31-37 Summary of the Invention [Means for solving the problem]

[0007] In some aspects, the disclosure relates to a method of administering a therapeutic agent to the subretinal space of a subject's eye, the method comprising the steps of creating a localized retinal detachment in the subretinal space of the subject's eye, injecting a composition comprising a magnetized therapeutic agent and a volume of a pharma- ceutically acceptable carrier, and applying a magnetic force to the eye, the magnetic force adhering the magnetized therapeutic agent to the subretinal space of the eye. In some embodiments, the process additionally requires removing at least 10%, 20%, 30%, 40%, or 50% of the volume of the pharma- ceutically acceptable carrier from the eye while the magnetic force is being applied to the eye.

[0008]

[0006] In another aspect, the disclosure relates to a method of treating a retinal disorder in a subject, the method comprising: creating a localized retinal detachment in the subretinal space of an eye of the subject; injecting a composition comprising a magnetization therapeutic agent and a volume of a pharma- ceutically acceptable carrier; and applying a magnetic force to the eye, the magnetic force adhering the magnetization therapeutic agent to the subretinal space of the eye. In certain embodiments, the method further comprises removing at least 10%, 20%, 30%, 40%, 50%, or 60% of the volume of the pharma- ceutical acceptable carrier from the eye while the magnetic force is applied to the eye. For example, in certain exemplary embodiments, 80-99% of the volume of the pharma- ceutical acceptable carrier is removed from the eye while the magnetic force is applied to the eye.

[0009]

[0007] In certain embodiments, the disclosure relates to the use of (i) a composition comprising a magnetotherapy agent and a pharma- ceutically acceptable carrier, and (ii) a magnetic force to guide and adhere the composition to the subretinal space of the eye, to treat a condition or disease, such as endothelial dystrophy, glaucoma, ischemic optic neuropathy, myopic degeneration, Leber's congenital amaurosis, retinitis pigmentosa, diabetic macular edema, proliferative diabetic retinopathy, retinopathy of prematurity, macular degeneration, age-related macular degeneration, or a combination thereof.

[0010]

[0008] In certain aspects, the magnetized therapeutic agent comprises magnetic particles and a therapeutic agent, the magnetic particles being attached to the therapeutic agent, and the therapeutic agent being selected from the group consisting of a gene therapy agent, an ocular cell, and a therapeutic drug.

[0011]

[0009] In certain methods, the ocular cells are selected from the group consisting of retinal pigment epithelial cells, photoreceptor cells, bipolar cells, ganglion cells, horizontal cells, amacrine cells, stem cells, retinal progenitor cells, and stem cell-derived ocular cells.

[0012]

[0010] Some methods further comprise the step of ceasing application of the magnetic force to the eye once at least 50% of the volume of the pharma- ceutically acceptable carrier has been removed.

[0013]

[0011] In certain embodiments, the magnetotherapy agent is a magnetized ocular cell, and application of magnetic force to the eye is stopped when the magnetized ocular cell is localized to Bruch's membrane, or to the retinal pigment epithelium layer, or to the posterior layer of the neural retina.

[0014]

[0012] Some embodiments further include the steps of injecting a second composition comprising a second magnetization therapeutic agent and a volume of a second pharma- ceutically acceptable carrier into the subretinal space of the eye, applying a magnetic force to the eye, which magnetic force adheres the second magnetization therapeutic agent to the subretinal space of the eye, and removing at least 50% of the volume of the second pharma- ceutically acceptable carrier from the eye while the magnetic force is applied to the eye.

[0015] In some embodiments, the second magnetization therapeutic agent is a different magnetization therapeutic agent than the first magnetization therapeutic agent.

[0016] In certain embodiments, the first and second magnetotherapy agents comprise ocular cells.

[0017]

[0015] In some embodiments, the first magnetotherapy agent comprises retinal pigment epithelial cells and the second magnetotherapy agent comprises ocular cells selected from the group consisting of photoreceptor cells, bipolar cells, ganglion cells, horizontal cells, and amacrine cells.

[0018] In certain embodiments, between 80 and 99% of the volume of the second pharma- ceutically acceptable carrier is removed while the magnetic force is applied to the eye.

[0019]

[0017] Some methods further comprise the step of ceasing application of the magnetic force to the eye once at least 50% of the volume of the second pharma- ceutically acceptable carrier has been removed.

[0020]

[0018] A localized retinal detachment may be created in some embodiments by injecting a balanced salt solution into the subretinal space of the eye.

[0021] In some methods, a localized retinal detachment is created by injecting a composition comprising a magnetotherapy agent into the subretinal space of the eye.

[0022]

[0020] Certain embodiments further include the steps of removing a quantity of vitreous through the pars plana and creating an incision in the retina in the subretinal space, wherein a composition comprising magnetized ocular cells is injected through the retinal incision into the subretinal space of the eye.

[0023] In some embodiments, the magnetic force attracts the magnetotherapy agent towards the subretinal space of the eye.

[0024] In certain embodiments, the magnetic force is applied by placing a magnet behind the eye.

[0025] In some embodiments, the magnet is placed intrinsically within the subject, hi certain embodiments, the magnet is sewn into place.

[0026] In yet another embodiment, the magnet is placed external to the subject.

[0027] In some embodiments, the magnetic force is applied by an external 3D magnet.

[0028]

[0026] Certain magnetic particles have an average diameter of 1 micron or less, 500 nm or less, 200 nm or less, or 50 nm or less. Some magnetic particles have a diameter of 1 micron or less, 500 nm or less, or 200 nm or less.

[0029]

[0027] Some magnetic particles comprise iron in any ferromagnetic form. Certain magnetic particles have a surface coating. The surface coating can allow for attachment of antibodies, antibody fragments, chemical moieties, proteins, or sugar fragments that bind to therapeutic agents.

[0030]

[0028] These and other aspects, features, and advantages will be apparent from the description and drawings, and from the accompanying claims, if any.

[0031]

[0029] The drawings depict certain non-limiting embodiments and implementations. [Brief description of the drawings]

[0032] [Figure 1A] 1 illustrates one magnetic surgery device positioning a magnetic body behind a patient's eye. [Figure 1B] 1 presents a perspective view of one of a patient's eyes, shown without the context of the surrounding eye socket or other structures and tissues. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0033]

[0032] Accordingly, it is to be understood that the embodiments of the invention described herein are merely illustrative of the application of the principles of the invention. References to details of the illustrated embodiments are not intended to limit the scope of the appended claims, which themselves recite those features regarded as essential to the invention.

[0034]

[0033] Detailed aspects and applications of the disclosure are described below in the accompanying drawings and detailed description of the technology. Unless otherwise noted, the words and phrases in the specification and claims are to be given their plain, ordinary, and accustomed meanings to those skilled in the art to which they apply.

[0035]

[0034] In the following description, for the purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the various aspects of the disclosure. However, as will be understood by those skilled in the relevant art, embodiments of the technology disclosed herein may be practiced without these specific details. It should be noted that there are many different alternative configurations, devices, and technologies to which the disclosed technology can be applied. The full scope of the technology disclosed herein is not limited to the examples described below.

[0036]

[0035] The terms "exemplary" and "embodiment" or various forms of these terms are used herein to mean serving as an embodiment, example, or illustration. Any aspect or design described as "exemplary" or as an "embodiment" should not necessarily be construed as preferred or advantageous over other aspects or designs. Furthermore, the examples are provided for clarity and understanding only and are not intended to limit or constrain in any way the disclosed subject matter or relevant portions of this disclosure. It should be understood that numerous additional or alternative embodiments of differing scope could have been presented, but have been omitted for the sake of brevity.

[0037]

[0036] When a range of values ​​is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, it will be understood that by use of the antecedent "about," the particular value forms another embodiment. All ranges are inclusive and combinable.

[0038]

[0037] Throughout the description and claims of this specification, the words "comprise" and "include" and variations of these words, such as "comprises" and "comprised," mean "including, but not limited to," and are not intended to (and do not) exclude other elements.

[0039]

[0038] This specification includes detailed embodiments of the present disclosure as necessary. It should be understood that the disclosed embodiments are merely examples of the disclosure that can be embodied in various forms. Therefore, specific structural and functional details disclosed in this specification should not be interpreted as limitations, but merely as a basis for teaching those skilled in the art to employ the present disclosure. The following specific examples will enable the disclosure to be more fully understood. However, they are provided merely as guidance and do not imply any limitations.

[0040]

[0039] The present disclosure will be more readily understood by reference to the following detailed description in conjunction with the accompanying figures and examples, which form a part of this disclosure. It is to be understood that the present disclosure is not limited to the specific materials, devices, methods, applications, conditions, or parameters described and / or illustrated herein, and that the terminology used herein is for the purpose of describing specific embodiments by way of example only, and is not intended to limit the invention as set forth in the claims. As used herein, the term "plurality" means more than one. When a range of values ​​is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, it will be understood that the particular value forms another embodiment by use of "about." All ranges are inclusive and combinable.

[0041]

[0040] This disclosure, its aspects, and its implementations are not limited to the specific material types, components, methods, or other examples disclosed herein. It is contemplated that many additional material types, components, methods, and procedures known in the art may be used with specific implementations from this disclosure. Thus, for example, even if a specific implementation is disclosed, such implementations and implementation components may comprise any components, models, types, materials, versions, quantities, and / or the like known in the art for such systems and implementation components that are not inconsistent with the intended operation.

[0042]

[0041] While the present disclosure includes embodiments in many different forms, specific embodiments have been shown in the drawings and are described in detail herein, with the understanding that the disclosure should be considered as illustrative of the principles of the disclosed methods and systems, and that it is not intended to limit the broad aspects of the disclosed concepts to the embodiments depicted.

[0043]

[0042] All amounts, ratios, and percentages are by weight unless otherwise indicated. The singular articles "a," "an," and "the" are equivalent to "a," "one," and "the," respectively, and refer to one or more, unless otherwise indicated by the context of the specification. Thus, for example, reference to "a process" includes reference to one or more of such processes.

[0044]

[0043] The disclosure of a range includes the range itself as well as any subsumed therein, including the endpoints. For example, the disclosure of a range of 2.0 to 4.0 includes not only the range 2.0 to 4.0, but also 2.1, 2.3, 3.4, 3.5, and 4.0 individually, as well as any other numbers subsumed within the range. Furthermore, the disclosure of a range, for example, 2.0 to 4.0, also includes subsets such as 2.1 to 3.5, 2.3 to 3.4, 2.6 to 3.7, and 3.8 to 4.0, as well as any other subsets subsumed within the range. As used herein, "about," "approximately," and "substantially" mean within a percentage difference of 20%, 10%, 5%, 3%, 2%, or 1% or less. Similarly, the disclosure of a Markush group includes the entire group as well as any individual members and subgroups subsumed within the group.

[0045]

[0044] Described herein are methods of administering a therapeutic agent to the subretinal space of an eye in a subject and methods of treating a retinal disorder in a subject. The methods include creating a localized retinal detachment in the subretinal space of the eye of the subject and injecting a composition comprising a magnetized therapeutic agent and a volume of a pharma- ceutically acceptable carrier. A magnetic force is applied to the eye, the magnetic force adhering the magnetized therapeutic agent to the subretinal space of the eye. At least 50% of the volume of the pharma- ceutical acceptable carrier is removed from the eye while the magnetic force is applied to the eye. In some embodiments, 80-99% of the volume of the pharma- ceutical acceptable carrier is removed while the magnetic force is applied to the eye. In some embodiments, the methods further include ceasing application of the magnetic force to the eye once at least 50% of the volume of the pharma- ceutical acceptable carrier has been removed.

[0046] In some aspects, the magnetized therapeutic agent comprises magnetic particles and a therapeutic agent. The magnetic particles are affixed to the therapeutic agent, and the therapeutic agent is selected from the group consisting of a gene therapy agent, an ocular cell, and a therapeutic drug. In certain embodiments, the ocular cell is selected from the group consisting of a retinal pigment epithelial cell, a photoreceptor cell, a bipolar cell, a ganglion cell, a horizontal cell, an amacrine cell, and a stem cell derived ocular cell. In certain embodiments where the magnetized therapeutic agent is a magnetized ocular cell, application of a magnetic force to the eye is terminated once the magnetized ocular cell has localized to Bruch's membrane.

[0047]

[0046] The magnetic particles preferably have an average diameter of 1 micron or less, 500 nm or less, 200 nm or less, or 50 nm or less. In certain embodiments, the magnetic particles have a diameter of 1 micron or less, 500 nm or less, or 200 nm or less. In some aspects, the magnetic particles comprise iron in any ferromagnetic form. In certain embodiments, the magnetic particles have a surface coating. For example, the surface coating allows for the attachment of an antibody, antibody fragment, protein, or sugar fragment that binds to a therapeutic agent.

[0048] In some aspects, the magnetic force attracts the magnetized therapeutic agent towards the subretinal space of the eye. In certain embodiments, the magnetic force is applied by placing a magnet behind the eye. In some aspects, the magnet is placed intrinsically to the subject, for example, the magnet is sewn into place. In other aspects, the magnet is placed externally to the subject. For example, the magnetic force is applied by an external 3D magnet.

[0049]

[0048] In certain embodiments of the methods described herein, the method further comprises injecting a second composition comprising a second magnetization therapeutic agent and a volume of a second pharma- ceutically acceptable carrier into the subretinal space of the eye, and applying a magnetic force to the eye, the magnetic force adhering the second magnetization therapeutic agent to the subretinal space of the eye. During application of the magnetic force to the eye, at least 50% of the volume of the second pharma- ceutically acceptable carrier is removed from the eye. In some embodiments, during application of the magnetic force to the eye, 80-99% of the volume of the second pharma- ceutically acceptable carrier is removed from the eye. In some embodiments, the method comprises ceasing application of the magnetic force to the eye once at least 50% of the volume of the second pharma- ceutically acceptable carrier has been removed.

[0050]

[0049] In some aspects, the second magnetization therapy agent is a different magnetization therapy agent than the first magnetization therapy agent. In some embodiments, the first magnetization therapy agent and the second magnetization therapy agent comprise ocular cells. For example, the first magnetization therapy agent comprises retinal pigment epithelial cells and the second magnetization therapy agent comprises ocular cells selected from the group consisting of photoreceptor cells, bipolar cells, ganglion cells, horizontal cells, and amacrine cells.

[0051] In certain embodiments of the methods described herein, a localized retinal detachment is created by injecting a balanced salt solution into the subretinal space of the eye. In other embodiments, a localized retinal detachment is created by injecting a composition comprising a magnetotherapy agent into the subretinal space of the eye.

[0052] In certain embodiments of the methods described herein, the method further comprises removing a volume of vitreous through the pars plana and creating an incision in the retina in the subretinal space. A composition comprising magnetized ocular cells is injected through the retinal incision into the subretinal space of the eye.

[0053]

[0052] The term "incision" may also include injection, puncture, retinotomy, or alternatively, the subretinal space may be accessed by puncturing through the suprachoroidal space, across the choroid and Bruch's membrane, and into the subretinal space. In this case, the magnetic force tends to draw the therapeutic agent away from the incision / puncture site rather than "towards the subretinal space." To facilitate removal of some or all of the carrier, the magnetic force may draw the therapeutic agent away from the incision site.

[0054]

[0053] The term "subject" refers to animals, such as mammals, particularly humans.

[0055]

[0054] The term "ocular disorder" refers to a condition or disease that interferes with the eye's ability to function properly and / or adversely affects the eye's visual clarity.

[0056]

[0055] The term "gene therapy" refers to a therapeutic technique in which a subject's genes are modified to treat or cure a disease.

[0057]

[0056] The term "pharmaceutical acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid, diluent, excipient, or solvent, involved in carrying or transporting a therapeutic composition in a subject. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the subject to which it is administered. Moreover, an acceptable carrier must not alter the specific activity of the therapeutic agent.

[0058]

[0057] The term "target tissue" refers to any particular tissue type or location, e.g., a location within an organ or tissue, to which it is desirable to deliver a therapeutic agent. For example, a therapeutic agent may be delivered to the eye or a particular area of ​​the eye, e.g., the cornea, optic nerve, retina, etc.

[0059]

[0058] The term "magnetic nanoparticles" refers to particles on the nanometer scale, ie, 1-500 nm, that have magnetic properties.

[0060]

[0059] Described herein is a method for delivering therapeutic agents to the subretinal space of the eye, which also relates to a method for treating retinal disorders. Many ophthalmic therapies are delivered locally in the form of eye drops or gels, but topical administration does not provide precise and efficient delivery. In order to deliver therapeutic agents to the subretinal space, several barriers must be overcome for effective delivery. Precorneal dynamic and static ocular barriers all limit the delivery of therapeutic agents to ocular tissues. In addition, ocular tissues do not adequately maintain therapeutic drug levels over time. Although injections are a common form of ocular therapeutic delivery that overcomes many of these barriers, repeated eye punctures are associated with endophthalmitis, bleeding, and retinal detachment, and are not well tolerated by patients. For cell-based therapies, injection of fluid into the subretinal space requires the formation of a bleb until the additional fluid is absorbed over time. However, the presence of a bleb can cause scar tissue to form in the eye that can adversely affect vision. The described methods of delivering therapeutic agents to the subretinal space of the eye address these limitations of existing therapies for treating ocular disorders and diseases. Examples of ocular disorders that can be treated with the methods described herein include, but are not limited to, degenerative vitreoretinal diseases such as retinitis pigmentosa, macular degeneration, and Leber's congenital amaurosis, age-related macular degeneration, diabetic macular edema, proliferative diabetic retinopathy, myopic degeneration, and retinopathy of prematurity.

[0061]

[0060] The method of delivering a therapeutic agent to the subretinal space of an eye described herein comprises the steps of creating a localized retinal detachment in the subretinal space of a subject's eye, injecting a composition comprising a magnetized therapeutic agent and a volume of a pharma- ceutically acceptable carrier, applying a magnetic force to the eye, which magnetic force adheres the magnetized therapeutic agent to the subretinal space of the eye, and removing at least 50% of the volume of the pharma- ceutical acceptable carrier from the eye while the magnetic force is applied to the eye. The localized retinal detachment may be created by methods established in the art, for example, by injecting a solution into the subretinal space. The solution may be a balanced salt solution or a composition comprising the magnetized therapeutic agent. In some embodiments, the step of creating the localized retinal detachment comprises removing a volume of vitreous through the pars plana and creating an incision in the retina in the subretinal space, and a composition comprising a magnetized therapeutic agent is injected through the retinal incision into the subretinal space of the eye.

[0062]

[0061] Pharmaceutical carriers suitable for application to the eye are known in the art, for example, those described in Remington: The Science and Practice of Pharmacy, 21st Ed. (2005). In some embodiments, at least 50% of the volume of the pharma- ceutically acceptable carrier is removed from the eye at least 1 minute, at least 2 minutes, at least 3 minutes, at least 4 minutes, at least 5 minutes, at least 10 minutes, at least 15 minutes, at least 20 minutes, at least 25 minutes, at least 30 minutes, at least 1 hour after injection of a composition comprising a magnetized therapeutic agent and a volume of a pharma- ceutically acceptable carrier. In certain embodiments, the pharma- ceutically acceptable carrier is removed from the eye within 3 hours, within 2 hours, or within 1 hour after injection of a composition comprising a magnetized therapeutic agent and a volume of a pharma- ceutical acceptable carrier.

[0063]

[0062] In certain embodiments, about 5 to 30 minutes after injection of a composition comprising a magnetic therapeutic agent and a volume of a pharma- ceutically acceptable carrier, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the volume of the pharma- ceutical acceptable carrier is removed from the eye. At least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the volume of the pharma- ceutically acceptable carrier is removed 5-10 minutes after injection, 5-15 minutes after injection, 5-20 minutes after injection, 5-25 minutes after injection, 5-30 minutes after injection, 10-15 minutes after injection, 10-20 minutes after injection, 10-25 minutes after injection, 10-30 minutes after injection, 15-20 minutes after injection, 15-25 minutes after injection, 15-30 minutes after injection, 20-25 minutes after injection, 20-30 minutes after injection, or 25-30 minutes after injection. Thus, in some embodiments, following injection of a composition comprising a magnetization therapeutic agent and a volume of a pharma- ceutically acceptable carrier, between 50% and 99% of the volume (e.g., 50-60%, 50-70%, 50-80%, 50-90%, 50-95%, 50-99%, 60-70%, 60-80%, 60-90%, 60-95%, 60-99%, 70-80%, 70-90%, 70-95%, 70-99%, 80-90%, 80-95%, 80-99%, 90-95%, 90-99%, 95-99%, or any other range between 50-99%) is removed or expelled. In another embodiment, following injection of a composition comprising a magnetization therapeutic agent and a volume of a pharma- ceutically acceptable carrier, no more than 99% of the volume of the pharma- ceutically acceptable carrier is expelled.

[0064] In certain embodiments, the magnetic force is applied by a magnet positioned behind the patient's eye. In other embodiments, the magnetic force is applied by a magnet coupled to the patient behind the retina, posterior to the sclera. The magnet can be coupled to the patient by stitches or any other acceptable coupling method. In some aspects, the magnet is positioned behind the patient's eye in a reversible manner so that the magnet can be removed when magnetic force is no longer needed.

[0065]

[0064] Figure 1A depicts a magnetic surgical device 135 positioning a magnetic body 140 behind a patient's eye 10 where no magnet was previously placed. The magnetic body 140 is shown being placed by an arm 160 of the magnetic surgical device 135 to which the magnetic body 140 is coupled, which extends through an incision in the skin and around a desired portion of the patient's eye 10. The arm 160 and magnetic body 140 can be steered and positioned by a handle 150 coupled to the arm 160. Figure 1A shows additional features of the eye 10, including the ciliary body 30, the conjunctiva 40, the anterior chamber 50, the cornea 60, the iris 70, the lens 80, the orbit 90, the optic nerve 100, the retina 110, the choroid 120, and the sclera 130.

[0066]

[0065] Figure 1B is a perspective view of a patient's eye 10 shown without the context of the surrounding eye socket or other structures and tissues. Similar to Figure 1A, Figure 1B shows (but with a larger magnification) a magnetic surgical device 135 positioning a magnetic body 140 at the back or posterior portion of the patient's eye 10, such as over the macula or in areas where retinal cells have deteriorated, been damaged, or are otherwise not functioning properly and are to be replaced or augmented with new magnetic stem cells 200. Figure 1B further shows a catheter 170 that may be used to place new magnetic stem cells at or near the site of the damaged retina. In some embodiments, the catheter 170 has a connection to a pump 210. Figure 1B also shows a sclerotomy and choroidal fistula 220, a subretinal cannulation 230, and a Healon lifter retina 240. The magnetic body may be positioned prior to placement of the new magnetic cells 200 to aid in positioning and alignment of the magnetic cells 200. As shown in FIG. 1B, the magnetic body 140 may create a magnetic field 180 that draws the magnetic cells 200 to a desired location. The size, shape, location, and orientation of the magnetic body 140 may be configured to match or approximate the size, shape, location, and orientation of the damaged portion of the eye 10 and of the area where the new magnetic cells 200 are to be positioned.

[0067]

[0066] Rather than applying the magnet externally, such as by centering the magnet over the cornea 10 and applying it as a patch to the surface of the eye 10 from outside the eyelid (thus pushing or repelling the magnetic cells 200), the magnetic material 140 is placed inside the patient and adjacent to the area to be treated, such as within the patient's eye socket or at the back of the eye 10 (so as to attract or draw the magnetic cells 200 to the desired location).

[0068]

[0067] If the magnetic body 140 is configured to matably couple with other tissue structures, the matable surfaces of the magnetic body 140 may be adapted accordingly. Figure 1A also shows a number of attachment devices 135 coupled to the magnetic body 140. There may be two, three, four, several dozen, one to a hundred, or any desired number of attachment devices that include loops, rings, or openings configured to receive stitched sutures for coupling the magnetic body 140 to a patient.

[0069]

[0068] In some embodiments, the magnet is left in place for a desired length of time during which the cells can begin to adhere, bind, attach, or grow into place and / or the therapeutic agent can be delivered to and interact with the appropriate tissue. For example, the magnet may be left in place for 5 minutes to 7 days, and in some cases more typically 1 hour to 3 days. The magnetic field 180 may assist in directing the magnetic therapeutic agent to a desired location in the subretinal space. In certain embodiments where the therapeutic agent is magnetic nanoparticle-bound cells, the magnetic field 180 may direct the cells to a particular location where the nanoparticle-bound cells (e.g., photoreceptor cells or retinal pigment epithelial cells or stem cells, etc.) attach to the vitreous surface or subretinal Bruch's membrane of the host / patient, after which natural cell attachment has occurred, the magnetic field 180 may be removed and the intrinsic magnet may be removed. In certain implementations, application of the magnetic field to the eye 10 is stopped once an appropriate volume of the pharma- ceutical acceptable carrier has been removed.

[0070]

[0069] The fixed / rare earth magnet, electromagnet, or superconducting magnet may provide sufficient magnetic field density uniformity and magnetic field 180 gradient to direct the cells and hold them in place. In some embodiments, the magnet can be turned on and off as magnetic force is required when positioned at the appropriate location behind the eye 10. The details of the magnetic field 180 strength may vary depending on the needs, so a stronger magnetic field / gradient may be used when the magnet needs to act over a larger distance over a larger area or surface of the damaged cells, and a weaker magnetic field / gradient may be used when the magnet can be localized closer to the implanted particles and / or target tissue. The magnetic therapeutic agent may be directed to the target tissue using a first magnetic field of a first strength, and the magnetic field 180 may then be modulated to refine the movement of the therapeutic agent and shape the tissue (or thereabouts). Rather than using a strong first magnetic field to move the therapeutic agent, the magnetic field 180 may be modulated to be a second magnetic field of a second strength weaker than the first strength to simply hold the magnetic therapeutic agent in the desired location until the desired adhesion or therapeutic delivery is achieved.

[0071]

[0070] The magnetic therapeutic agent is a therapeutic agent magnetized with magnetic nanoparticles. The magnetic nanoparticles have an average diameter of about 500 nm or less, more commonly 200 nm or less, but preferably 100 nm or less. Particles that can be used include nanospheres, conjugates, micelles, colloids, aggregates, and complexes comprising ferromagnetic, paramagnetic, or superparamagnetic materials such as iron, nickel, cobalt, and their alloys that are considered suitable for in vivo use. For example, the magnetic nanoparticles may comprise any ferromagnetic form of iron, with or without an inert surface coating, the surface of which is chemically modified to allow for the attachment of antibodies, or antibody fragments, or proteins, or sugar fragments that bind to cells. As will be appreciated by those skilled in the art, compounds that are excessively toxic when used in accordance with the method should be avoided. In many applications, the particles are endocytosed and excreted over time, and it is therefore expected that small amounts of potentially toxic particles will not cause problems when used in the method.

[0072]

[0071] In general, magnetic nanoparticles will have a diameter of 5-500 nm, more specifically 40-400 nm, and most specifically 40-100 nm. Difficulties in using nanoparticles compared to micron-scale particles include particle aggregation, particle tracking and observation, and the ability to mobilize particles by an external magnetic field 180, all of which are much easier when using micron-scale or larger particles. For this reason, previous efforts have tended to focus on micron-scale particles, and it appears that the advantages of using nanoparticles compared to micron-scale particles have been ignored. The advantages include the ability to bind to cell surfaces without stimulating endocytosis, the ability to shed from cell surfaces or be excreted from cells when internalized, and the ability to be excreted from the eye 10 or body when shed from cells.

[0073] Various forms of magnetic nanoparticles have already been used in clinical and research applications without any demonstrated toxicity. For example, superparamagnetic particles containing microcrystalline iron oxide nanoparticles (MIONs) with diameters of <50 nm have been used as MRI contrast agents. These particles have demonstrated neurological non-toxicity and axonal transport of iron-based agents (Neuwell et al., Neurosurgery. 1994, 34: 777-784). Published studies supporting the use of the MRI contrast agent Ferridex (Advanced Magnetics and Berlex Laboratories) have found no adverse effects. Furthermore, magnetically directed drug delivery using labeled pharmaceuticals in the form of magnetic microspheres and magnetic polymer carriers has shown success in delivering antitumor drugs and radioisotypes to magnetically targeted areas in vivo (Schutt et al., Hybridoma. 1997, 16: 109-117; Lubbe et al., J Surg Res. 2001, 95: 200-206). In certain embodiments, the magnetic nanoparticles are those described in Miltenyi et al., Cytometry. 1990, 11:231-238. For example, the magnetic nanoparticles are dextran coated and have a diameter of 30±20 nm (measured by electron microscopy) or 65±20 nm (measured by dynamic light scattering). In a particular embodiment, the magnetic nanoparticles are CliniMACS® by Miltenyi Biotec with a diameter of 50 nm.

[0074] In some embodiments, the magnetic particles are coated. The coating applied to the magnetic particles includes a non-specific binder, such as an inert metal or polymer, such as gold or dextran, and / or a specific binder, such as an antibody specific for a cell surface antigen. For example, antibodies against SSEA-1 bind to many types of stem cells, and nanoparticles coated with anti-SSEA-1 antibodies can be used to convert stem cells into magnetic stem cells. Similarly, many cells express specific surface receptors, and antibodies against these specific receptors conjugated to magnetic nanoparticles bind to these cells, such as corneal endothelial cells, creating magnetic corneal endothelial cells. The coatings are applied to the particles by standard methods commonly used in the art (e.g., Schroder et al., J Immunol Methods. 1986, 93: 45-53; Douglas et al., Crit Rev Ther Drug Carrier Syst. 1987, 3: 233-261; Sestier et al., Electrophoresis. 1998, 19: 1220-1226). (See Perrin et al., J Immunol Methods. 1999, 224:77-87; McCloskey et al., Cytometry. 2000, 40:307-315; Tibbe et al., Cytometry. 2001, 43:31-37).

[0075]

[0074] Magnetic nanoparticles can be endocytosed by cells or attached to the surface of cells by any effective means known to those skilled in the art. For example, magnetic particles can be attached to cells using antibodies, such as antibodies specific to surface antigens present on the cells. For example, surface coatings with anti-L1, anti-trkB, anti-integrin, and cholera toxin subunit B can be placed on the magnetic particles for the purpose of attaching the magnetic particles to retinal ganglion cells. Magnetic particles can also be attached to cells using specific ligands whose receptors are present on the cells. For example, magnetic particles can be functionalized for attachment to retinal ganglion cells using brain-derived neurotrophic factor (BDNF). Coated magnetic nanoparticles can also be attached to the outer surface of cells by co-incubation in a common medium that provides sufficient cell viability during the co-incubation period, although the medium has not generally been found to be closely related to the attachment process. Generally, a balanced salt solution with a physiological pH of about 7.4 will suffice, but supplementation of the media to enhance cell survival during the process is the topic of other published work specific to the cell type used and is not germane to this disclosure. The time and temperature of the co-incubation may depend on the particular cell type to be converted to magnetic cells, for example, binding to retinal ganglion cells using magnetic nanoparticles coated with anti-trkB antibodies occurs maximally after 4 hours at 37° C. but may be performed overnight at 4° C. Excess magnetic nanoparticles that fail to bind to the cells can be washed away by centrifuging the cells and spinning at a speed that pellets the cells but does not pellet the unbound magnetic nanoparticles, or by using a magnetic field 180 to elute the magnetic nanoparticle-bound cells from the unbound cells, or both.

[0076]

[0075] Other means of attaching magnetic particles to therapeutic agents include non-specific chemical modifications such as carboxy or amide groups, or coating with a sugar or dextran, or a polymer such as an amino acid polymer like polylysine, or coating with another inert coating that binds to the therapeutic agent.

[0077] In yet another aspect, the magnetized therapeutic agent is an ocular cell, such as a retinal cell (retinal Müller glial cell or retinal astrocyte or other), a retinal endothelial cell or pericyte, a retinal progenitor cell, a retinal stem cell, an optic nerve glial cell (whether an astrocyte, an oligodendrocyte, a microglial cell, or a precursor thereof), or other stem or progenitor cell capable of differentiating into an ocular cell or an optic nerve cell or supporting the survival or growth or normal function of an ocular cell or an optic nerve cell. In some embodiments, the ocular cell is selected from the group consisting of a retinal pigment epithelial cell, a photoreceptor cell, a bipolar cell, a ganglion cell, a horizontal cell, an amacrine cell, and a stem cell. In some aspects, the magnetized ocular cell is a cell coated with magnetic nanoparticles. Also provided are normal or genetically modified cells that have endocytosed, bound, or affixed magnetic nanoparticles to their surface, either covalently or by antibody-antigen binding.

[0078]

[0077] The cells may be suspended in any pharma- ceutically / physiologically acceptable medium or solution, such as isotonic saline, culture medium, or a transport medium suitable for in vivo delivery to a subject. Additional excipients and carriers may be added as found appropriate by those skilled in the art. Suitable solutions and delivery vehicles are described in Remington: The Science and Practice of Pharmacy, 21st Ed. (2005). In some applications, such as subretinal delivery of stem cell-derived cells for age-related macular degeneration or retinitis pigmentosa, 10 3 ~10 6 Cells will be delivered by injection in volumes ranging from 3 to 300 µL, but more commonly in volumes of 10 4 The cells are delivered by injection in a volume of 10-100 μL. In other applications, such as delivery of stem cells to enhance survival of retinal ganglion cells in diseases such as glaucoma or ischemic optic neuropathy, 10 3 ~10 6Cells are delivered by injection in volumes of 3-300 µL, but more typically in volumes of 10 5 of cells are delivered by injection in a volume of 200 μL. When considering the delivery of cells geared towards carrying toxic compounds to specific tissues, for example in cancer therapy, the cell number would need to be carefully titrated against systemic toxicity to the patient.

[0079]

[0078] In some aspects, the methods described herein comprise administering a second magnetic therapeutic agent after the first magnetic therapeutic agent has been delivered. The second magnetic therapeutic agent may be the same as the first magnetic therapeutic agent or may be different from the first magnetic therapeutic agent. Preferably, 50-99% of the volume of the pharma- ceutically acceptable carrier in the composition having the first magnetic therapeutic agent is removed from the eye 10 prior to injecting the composition comprising the second magnetic therapeutic agent and a volume of the second pharma- ceutically acceptable carrier.

[0080]

[0079] In an exemplary embodiment, the first magnetic therapeutic agent comprises magnetized ocular cells, and the second magnetic therapeutic agent also comprises magnetized ocular cells. In some aspects, the second magnetic therapeutic agent comprises a different type of ocular cells than the first magnetic therapeutic agent, for example, to reconstruct the retinal cellular structure or to reconstruct the retinal pigment epithelium. During administration of the first magnetic therapeutic agent, the magnetic field 180 may be modulated by activating different zones, portions, or areas of the magnet to promote migration of the cells to specific areas. Cell migration can be directed in real time while observing the placement of the cells, such as when the cells include visual markers and are visually observed during placement. The placement of the cells may be observed or monitored indirectly or in any other suitable manner. Once the cells have achieved the desired adhesion to the target tissue, the pharma- ceutical acceptable carrier is expelled. The time to desired adhesion can be as early as 1 or 5 minutes after injection or as long as 3 hours after injection, and one of skill in the art may fine-tune the time required for cell adhesion to the target tissue to occur and the amount and duration of the magnetic field required before the cells are sufficiently attached and able to expel the pharma- ceutically acceptable carrier. After the pharma- ceutical acceptable carrier from the first therapeutic agent has been expelled, the second therapeutic agent can be administered by the described method and the magnetic eye cells can be directed to the desired target tissue through magnetic forces. This method allows for multiple layers or stacks of cells to be placed with delays between each stacking step, allowing time for the first layer to adhere, and then adding a subsequent second (or any number or 1+nth) layer of cells. In some cases, the first and subsequent cell layers have the same or similar footprint, while in other cases the subsequent cell layers may have different footprints or contain different cell types. In such a manner, for applications in which cells are not drawn and held in place, the number of non-adherent cells can be reduced, thereby reducing the number of non-adherent cells that would otherwise die and remain free floating and away from the desired binding site.

[0081] Other combinations of a first magnetic therapeutic agent and a second magnetic therapeutic agent include, but are not limited to, the examples listed below, namely: the first magnetotherapy agent comprises ocular cells and the second magnetotherapy agent comprises a therapeutic drug; the first magnetotherapy agent comprises ocular cells and the second magnetotherapy agent comprises a gene therapy agent; the first magnetization therapeutic agent comprises a first therapeutic drug and the second magnetization therapeutic agent comprises a second therapeutic drug; the first magnetotherapy agent comprises a therapeutic drug and the second magnetotherapy agent comprises eye cells; the first magnetic therapeutic agent comprises a therapeutic drug and the second magnetic therapeutic agent comprises a gene therapeutic agent; the first magnetic therapeutic agent comprises a gene therapeutic agent and the second magnetic therapeutic agent comprises a therapeutic drug; the first magnetic therapeutic agent comprises a gene therapeutic agent and the second magnetic therapeutic agent comprises ocular cells, or Includes embodiments in which the first magnetic therapy agent comprises a gene therapy agent and the second magnetic therapy agent comprises a gene therapy agent.

[0082]

[0081] In yet another embodiment, the method further comprises administering a third, fourth, fifth or sixth magnetization therapeutic agent following the administration of the first and second magnetization therapeutic agents and the expulsion of their respective pharma- ceutically acceptable carriers. In such an embodiment, application of magnetic force to the eye 10 is stopped when 50-99% of the volume of the pharma- ceutically acceptable carrier in the composition of the last administered magnetization therapeutic agent has been removed from the eye 10.

[0083]

[0082] It is to be understood that the disclosed embodiments are not limited to the specific components disclosed herein, and that virtually any components may be utilized that are not inconsistent with the intended operation of the method and / or system implementations for such embodiments. Thus, for example, while examples of specific components may be disclosed, such components may be configured in any shape, size, style, type, model, version, class, grade, measure, density, material, weight, quantity, and / or the like that are not inconsistent with the intended purpose, method, and / or system of the implementation. Where the above description refers to a particular implementation or embodiment, it will be readily apparent that numerous modifications may be made without departing from the scope and / or spirit thereof, and that these principles and modifications may be applied to other such embodiments. The disclosed embodiments are therefore to be considered in all respects as illustrative and not restrictive.

[0084]

[0083] It is contemplated that many additional types of materials, components, methods, and procedures known in the art may be used with particular embodiments from this disclosure. Thus, for example, even if a particular embodiment is disclosed, such embodiments and implementation components may comprise any components, models, types, materials, versions, quantities, and / or the like known in the art for such systems and implementation components that are not inconsistent with the intended operation. Additional embodiments are possible and are covered by this disclosure. Thus, further embodiments and embodiments based on the teachings of this disclosure are within the scope of the following claims. [Explanation of symbols]

[0085] 10 eyes 30 Ciliary body 40 Conjunctiva 50 Anterior Chamber 60 Cornea 70 Iris 80 crystalline lens 90 Orbital 100 Optic Nerve 110 Retina 120 Choroid 130 Sclera 135 Magnetic surgery device, attachment device 140 Magnetic material 150 Handle 160 Arm 170 Catheter 180 Magnetic field 190 Sutures 200 new magnetic cells 210 Pump 220 Scleral incision and choroidal fistula 230 Subretinal Cannulation 240 Healon lifter retina

Claims

1. A pharmaceutical composition for treating a retinal disorder, comprising: (a) a magnetized therapeutic agent comprising a therapeutic agent and magnetic particles linked to, bound to, or internalized by the therapeutic agent; (b) a pharmaceutically acceptable carrier; the composition is formulated for subretinal administration to the subretinal space of the eye; A pharmaceutical composition, wherein when the composition is administered to the subretinal space and an external magnetic field is applied to the eye, at least 50% of the volume of the carrier is removed from the eye while the external magnetic field is applied, while the magnetization therapeutic agent is guided to and retained within the subretinal space at a target site.

2. A set of pharmaceutical compositions for use in treating retinal disorders, comprising: (i) a first pharmaceutical composition comprising a first magnetization therapeutic agent and a first pharmaceutically acceptable carrier; (ii) a second pharmaceutical composition comprising a second magnetization therapeutic agent and a second pharmaceutically acceptable carrier; During use in treating a retinal disorder, the first pharmaceutical composition is administered to the subretinal space of an eye and a magnetic force is applied to the eye, and after administration of the first pharmaceutical composition, at least 50% of the volume of the first pharmaceutically acceptable carrier of the first pharmaceutical composition is removed from the eye while the magnetic force is applied, and then the second pharmaceutical composition is administered to the subretinal space.

3. In the pharmaceutical composition according to claim 1 or the set according to claim 2, The pharmaceutical composition or set, wherein the external magnetic field is applied by a magnetic body positioned behind the eye.

4. 10. The pharmaceutical composition of claim 1, the carrier is a pharmaceutical composition that is removed by aspiration of carrier fluid from the subretinal space while the external magnetic field is applied; or 3. The set according to claim 2, The first pharmaceutically acceptable carrier is removed from the subretinal space by aspiration of carrier fluid while the external magnetic field is applied.

5. 10. The pharmaceutical composition of claim 1, a pharmaceutical composition, wherein at least 80% of the volume of the carrier is removed from the eye while the external magnetic field is applied; or 3. The set according to claim 2, At least 80% of the volume of the first pharmaceutically acceptable carrier is removed from the eye while the external magnetic field is applied, and then the second pharmaceutical composition is administered to the subretinal space.

6. 10. The pharmaceutical composition of claim 1, wherein the magnetic particles have an average diameter of 500 nm or less, 200 nm or less, or 50 nm or less.

7. 10. The pharmaceutical composition of claim 1, the magnetic therapeutic agent is a gene therapy agent, a pharmaceutical composition, or 3. The set according to claim 2, A set, wherein the first magnetization therapeutic agent is a gene therapy agent.

8. 10. The pharmaceutical composition of claim 1, the magnetic therapeutic agent is an ocular cell, or a pharmaceutical composition; 3. The set according to claim 2, A set, wherein the first magnetotherapy agent is an ocular cell.

9. 9. The pharmaceutical composition or set according to claim 8, A pharmaceutical composition or set, wherein the ocular cells are selected from the group consisting of retinal pigment epithelial cells, photoreceptor cells, bipolar cells, ganglion cells, horizontal cells, amacrine cells, stem cells, retinal progenitor cells, and stem cell-derived ocular cells.

10. In the pharmaceutical composition according to claim 1 or the set according to claim 2, The pharmaceutical composition or set, wherein the retinal disorder is selected from retinitis pigmentosa, Leber's congenital amaurosis, age-related macular degeneration, diabetic macular edema, proliferative diabetic retinopathy, myopic macular degeneration, and retinopathy of prematurity.

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