Ultra-low dose zoledronic acid for treating retinal diseases

Abstract

Methods of treating retinal diseases using low doses of bisphosphonate inhibitors of acid sphingomyelinase (ASM) and farnesyl diphosphate synthase (FDPS), such as zoledronic acid, are provided herein. TIFF2025517691000003.tif78128

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 340,435, filed May 10, 2022, and U.S. Provisional Patent Application No. 63 / 500,448, filed May 5, 2023, each of which is hereby incorporated by reference in its entirety for all purposes.

Background Art

[0002] Background of the Invention The retinal pigment epithelium (RPE) is an essential component of the eye. The RPE is composed of flat, mosaic - shaped hexagonal cells that are tightly joined at their junctions. The RPE contacts the sensory retinal cells on one side, which sense light and transmit visual information to the optic nerve. The RPE contacts the choroid tissue on the other side, which is a vascular region that supplies water, nutrients, and other compounds to the cells above it in the eye. The RPE plays many important roles in the maintenance of vision, including isolating the eye tissues from the systemic circulatory system, maintaining an appropriate ionic environment, processing the outer segment elements of photoreceptor cells that have detached from the neural retina, and protecting the retina from excessive light. Thus, the RPE is essential for vision because it maintains the photoreceptor cells it supports.

[0003] Various retinal diseases can have an adverse impact on this important eye component. The main lesion of the RPE is dry age-related macular degeneration (AMD), which is a disease in which vision is gradually impaired in the macula, the central region of the eye. AMD is one of the main causes of vision loss in people over 60 years old. It is estimated that 30% of people over 75 years old in the United States suffer from some form of AMD. In dry AMD, the accumulation of lipofuscin bisretinoids is observed. These species are vitamin A metabolites that include the tips of undegraded photoreceptor outer segments, which are normally degraded in the RPE. These lipofuscin bisretinoids gradually accumulate as a byproduct of the steady recycling of the retinal chromophore. Another symptom of dry AMD is the accumulation of lipid-protein aggregates called drusen above and below the RPE, which interfere with the contact with the choroid that provides nutrients and is located beneath the RPE. The progressive accumulation of lipofuscin, bisretinoids, and drusen is associated with the dysfunction and death of RPE cells.

[0004] Other retinal pathologies that are severe and lead to blindness and are associated with the accumulation of lipofuscin are also known and afflict millions of subjects. For example, Stargardt macular dystrophy, including autosomal dominant Stargardt disease or autosomal recessive Stargardt disease, is a pathology characterized by the accumulation of lipofuscin and macular degeneration. Other retinal pathologies include, for example, neuronal ceroid lipofuscinosis, including Batten disease, and Best vitelliform macular dystrophy.

[0005] Previous studies have elucidated the pathological cascades underlying dry-type AMD and other retinal pathologies associated with the accumulation of lipofuscin, as described, for example, in Toops et al., 2015. Cholesterol-mediated activation of acid sphingomyelinase disrupts autophagy in the retinal pigment epithelium. Mol. Biol. Cell 26: 1-14 (Non-Patent Document 1). These studies have determined that bisretinoids sequester cholesterol and bis(monoacylglycero)phosphate, a cofactor for acid sphingomyelinase (ASM), within the RPE. This has been further demonstrated to promote the activation of ASM, and this activation in turn increases the accumulation of ceramide. Ceramide promotes the acetylation of microtubules, and this disrupts the normal transport of autophagosomes and negatively impacts the essential autophagic flux in the RPE. Inhibition of ASM has been further demonstrated to restore efficient autophagy and enhance the integrity of the RPE by interfering with the pathological cascade initiated by lipofuscin bisretinoids.

[0006] Therapeutic interventions based on the above findings are taught, for example, in U.S. Patent Application Publication No. 20150366876 (Patent Document 1) by Lakkaraju et al., Use of Inhibitors of Acid Sphingomyelinase to Treat Acquired and Inherited Retinal Degenerations. In the above, it has been demonstrated that desipramine, a functional ASM inhibitor, can effectively inhibit the accumulation of ceramide and the pathological cascade thereby initiated. In addition, the therapeutic use of other ASM-inhibiting agents, including zoledronic acid, a structural ASM inhibitor, has been suggested but not demonstrated.

[0007] Despite these recent promising advances in understanding and treating retinopathies, clinical applications are still not available, and millions of people worldwide continue to suffer from retinopathies. Thus, there is a great need in the art for effective therapies for treating retinopathies, such as dry AMD, Stargardt's disease, and the like.

[0008] As suggested by previous studies, ASM inhibitors other than desipramine may also be potentially useful in treating retinopathies. Zoledronic acid is a known ASM inhibitor and thus may be potentially useful in treating retinopathies. However, to the inventors' knowledge, the evaluation of this agent and related bisphosphonate-based ASM inhibitors with respect to retinal diseases has not been performed heretofore. Zoledronic acid also inhibits farnesyl diphosphate synthase (FDPS), which is an enzyme involved in cholesterol biosynthesis and protein prenylation.

[0009] Zoledronic acid, (1-hydroxy-2-imidazol-1-yl-phosphonoethyl) phosphonic acid monohydrate, also known as zoledronate, is a bisphosphonate that includes TIFF2025517691000002.tif36128. Among its various biological actions, zoledronic acid is an inhibitor of ASM, as described above (Roth, et al., Angew Chem Int Ed Engl. 48:7560-7563, 2009 (Non-Patent Document 2)), and is also an inhibitor of farnesyl diphosphate synthase (FDPS).

[0010] Zoledronic acid also inhibits bone resorption by inhibiting osteoclastic activity and inducing apoptosis of osteoclasts. Zoledronic acid also binds to bone and blocks osteoclastic resorption of mineralized bone. Clinically, zoledronic acid is approved for the treatment of osteoporosis and is sold in various forms, such as Aclasta (trademark) (Novartis Pharmaceuticals) and Reclast (trademark) (Novartis Pharmaceuticals). Zoledronic acid is also approved for use in the treatment of Paget's disease.

[0011] Regarding cancer, zoledronic acid is also approved for use in the treatment of skeletal complications resulting from some cancers, for example, for use in the treatment of hypercalcemia associated with malignancy. Zoledronic acid is also capable of inducing apoptosis in cancer cells and is approved for the treatment of multiple myeloma and for the treatment of bone metastases from some solid tumors. In addition, some types of solid tumors (breast cancer, prostate cancer, lung cancer) can also metastasize to the bone marrow. These cancers disrupt the function of the osteoclasts present in the bone marrow, promoting pathological bone resorption and inhibiting the formation of new bone. Zoledronic acid, by inhibiting osteoclasts and bone resorption and inducing apoptosis in cancer cells, is approved for the treatment of those lesions and is sold as Zometa (trademark) (Novartis Pharmaceuticals).

[0012] Generally, zoledronic acid is considered to be a safe and well-tolerated agent. However, although zoledronic acid has demonstrable therapeutic utility, this agent is also associated with a variety of negative side effects. Various sources have pointed out side effects of zoledronic acid such as anemia, fatigue, muscle discomfort, and swelling of the lower extremities. Furthermore, in some subjects, the use of zoledronic acid can pose a risk of renal insufficiency, and its use may not be recommended for subjects with sub-normal renal function, such as subjects with CKD. A rare but serious complication in some subjects treated with bisphosphonates is osteonecrosis of the jaw, which occurs primarily in subjects with multiple myeloma who have undergone tooth extraction. In addition, the European Medicines Agency has reported that atypical fractures may be a side effect of bisphosphonates. Furthermore, subjects with hypercalcemia may be at risk of adverse side effects of zoledronic acid.

[0013] As with many therapeutic agents, the administration of bisphosphonates also increases the risk of unfavorable interactions with other medications being administered to the subject. For example, an increased risk of gastrointestinal bleeding results from unfavorable interactions between zoledronic acid and drugs such as aspirin, celecoxib, etc. An increased risk of nephrotoxicity has been observed resulting from unfavorable interactions between zoledronic acid and drugs such as acyclovir and cisplatin.

Prior Art Documents

Patent Documents

[0014]

Patent Document 1

Non-Patent Documents

[0015]

Non-Patent Document 1

[0016] Brief Summary of the Invention The invention disclosed herein is based on the unexpected discovery that bisphosphonate-based ASM / FDPS inhibitors can be effectively administered at ultra-low doses, not intended or suggested from their previous uses, to treat retinal diseases. Specifically, the inventors of the present disclosure have identified that zoledronic acid and related compositions can be therapeutically effective at doses significantly lower than their standard doses currently used clinically, for example at least 1 / 100. In particular, the dose is approximately 1 / 2,000 compared to the dose of desipramine that has previously been shown to achieve a therapeutic effect in the retina.

[0017] The surprising and unexpected efficacy of these agents at ultra-low doses provides various benefits. In a first aspect, by using at ultra-low doses, bisphosphonates, such as zoledronic acid, can be used to treat retinal pathologies, where the risk of harmful side effects, or the risk of unwanted drug cross-interactions, observed when used at standard doses is substantially reduced. Such a reduction in risk substantially expands the pool of subjects with retinal diseases treatable by bisphosphonates. Such therapeutic and other therapeutic advantages are provided by the methods of the invention as disclosed herein.

[0018] In one aspect, the present disclosure features a method of treating a retinal disease in a subject in need thereof by administering a therapeutically effective amount of a pharmaceutical composition comprising zoledronic acid or a derivative thereof to the subject, wherein the zoledronic acid or derivative is administered at an ultra-low dose. In some embodiments, the retinal disease is a condition mediated by the accumulation of lipofuscin in RPE cells. In some embodiments, the retinal disease is dry age-related macular degeneration or Stargardt macular dystrophy. In some examples, the retinal disease is neuronal ceroid lipofuscinosis, Batten disease, Best vitelliform macular dystrophy, Niemann-Pick disease type C, Doin honeycomb dystrophy, Faber disease, or Best vitelliform macular dystrophy. In some embodiments, the pharmaceutical composition comprises an implant; a drug-eluting device, structure, or substance; a polymeric drug-eluting wafer; an injectable hydrogel; or an implantable hydrogel scaffold, or is incorporated therein. In some embodiments, the zoledronic acid or derivative is administered by an intravitreal implant to deliver a dose of about 50 ng / day to about 50 μg / day to one eye. In some embodiments, the zoledronic acid or derivative is administered to provide an intraocular concentration of zoledronic acid of at least 100 pM, 1 nM, 10 nM, 100 nM, 200 nM, 500 nM, or 1 μM. In some embodiments, the zoledronic acid or its derivative is administered as an eye drop solution or suspension, or as an ophthalmic ointment or gel, at a dose of about 50 ng / day to about 50 μg / day to one eye. In some embodiments, the zoledronic acid or derivative is administered by suprachoroidal injection at about 50 ng / day to about 50 μg / day to one eye. In some embodiments, the zoledronic acid or derivative is administered systemically to provide a dose of 0.001 to 2.0 mg, 0.01 to 2.0 mg, or 0.3 to 0.5 mg.In some embodiments, zoledronic acid or a derivative is administered systemically at a dose of 0.3 mg to 0.5 mg, and is administered at a dose selected from the group consisting of 0.001 mg, 0.02 mg, 0.03 mg, 0.04 mg, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.10 mg, 0.20 mg, 0.30 mg, 0.40 mg, 0.50 mg, 0.60 mg, 0.70 mg, 0.80 mg, 0.90 mg, and 1.0 mg. In some embodiments, zoledronic acid or a derivative is administered systemically at a dose of 100 ng to 10 μg per kg of body weight, at a dose of 1.0 to 7.0 μg per kg of body weight, or at a dose of about 5.0 μg per kg of body weight. In some embodiments, zoledronic acid or a derivative is administered systemically in an amount of 0.1 μg, 0.2 μg, 0.3 μg, 0.5 μg, 1.0 μg, 2.0 μg, 3.0 μg, 4.0 μg, 5.0 μg, 6.0 μg, 7.0 μg, 8.0 μg, 9.0 μg, or 10.0 μg per kg of body weight. In some embodiments, administration is by a route including any of intravenous delivery, intramuscular delivery, intraperitoneal delivery, or subcutaneous delivery. In some embodiments, the pharmaceutical composition is administered at a frequency selected from the group consisting of once a year, once a month, twice a month, once a week, twice a week, every other day, once a day, twice a day, and three times a day. In some embodiments, the pharmaceutical composition comprises zoledronic acid and comprises any of an excipient, a carrier, a diluent, a release formulation, a drug delivery vehicle or a drug targeting vehicle, and an additional active therapeutic agent. In some embodiments, the pharmaceutical composition comprises an adiponectin 1 receptor agonist, such as adiporone. In some embodiments, zoledronic acid is co-administered with an adiponectin 1 receptor agonist.

[0019] In yet another aspect, the present disclosure provides an intravitreal implant comprising zoledronic acid, and the intravitreal implant is loaded with an amount of zoledronic acid of 0.001 to 0.3 mg. In some embodiments, the implant is loaded with 0.005 to 2.5 mg of zoledronic acid.

[0020] In another aspect, the present disclosure provides an ophthalmic topical preparation comprising zoledronic acid at a dosage of 0.001 to 0.05 mg / dosage. In some embodiments, the ophthalmic preparation comprises zoledronic acid at a dosage of 0.005 to 0.05 mg / dosage.

[0021] In yet another aspect, the present disclosure provides an injectable ophthalmic preparation comprising zoledronic acid at a concentration of 1 μg / ml to 10 mg / ml. In some embodiments, the injectable ophthalmic preparation comprises zoledronic acid at a concentration of 10 μg / ml to 1 mg / ml. In some embodiments, the ophthalmic preparation comprises zoledronic acid conjugated with a dendrimer, or zoledronic acid formulated as nanoparticles.

[0022] In yet another aspect, the present disclosure provides: a method of treating a retinal disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising adiporone; or a method of treating a retinal disease in a subject in need thereof by administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a bisphosphonate ASM / FDPS inhibitor, wherein the bisphosphonate ASM / FDPS inhibitor is administered at an ultra-low dose. In some embodiments, the retinal disease is a condition mediated by the accumulation of lipofuscin in RPE cells. In some embodiments, the retinal disease is Stargardt macular dystrophy, or dry age-related macular degeneration. In some embodiments, the retinal disease is neuronal ceroid lipofuscinosis, Batten disease, Best vitelliform macular dystrophy, Niemann-Pick disease type C, Doin honeycomb dystrophy, Faber disease, or Best vitelliform macular dystrophy. In some embodiments, adiporone, or the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.001 to 2.0 mg, at a dose of 0.1 to 1.0 mg, or at a dose of 0.3 to 0.5 mg. In some embodiments, adiporone, or the bisphosphonate ASM inhibitor is administered at a dose selected from the group consisting of 0.001 mg, 0.002 mg, 0.003 mg, 0.004 mg, 0.005 mg, 0.006 mg, 0.007 mg, 0.008 mg, 0.009 mg, 0.01, 0.02 mg, 0.03 mg, 0.04 mg, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.10 mg, 0.20 mg, 0.30 mg, 0.40 mg, 0.50 mg, 0.60 mg, 0.70 mg, 0.80 mg, 0.90 mg, and 1.0 mg. In some embodiments, adiporone, or the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 100 ng to 10 μg per kg of body weight, at a dose of 1.0 to 7.0 μg per kg of body weight, or at a dose of about 5.0 μg per kg of body weight.In some embodiments, adiponectin, or a bisphosphonate-based ASM / FDPS inhibitor, is administered at a dosage selected from the group consisting of 0.1 μg, 0.2 μg, 0.3 μg, 0.5 μg, 1.0 μg, 2.0 μg, 3.0 μg, 4.0 μg, 5.0 μg, 6.0 μg, 7.0 μg, 8.0 μg, 9.0 μg, and 10.0 μg per kg of body weight. In some embodiments, the pharmaceutical composition is administered at a frequency selected from the group consisting of once a year, once a month, twice a month, once a week, twice a week, every other day, once a day, twice a day, and three times a day. In some embodiments, administration is by a route including any of systemic delivery; local delivery; intravenous delivery; intramuscular delivery; intraperitoneal delivery; topical delivery; subcutaneous delivery; intravitreal delivery; suprachoroidal delivery, and topical delivery to the eye. In some embodiments, the pharmaceutical composition comprises one or more bisphosphonate-based ASM / FDPS inhibitors, or adiponectin, and comprises any of an excipient, a carrier, a diluent, a release formulation, a drug delivery vehicle or a drug targeting vehicle, and an additional active therapeutic agent.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0024] Detailed Description As used herein, "a", "an", and "the" include aspects in which there is one member, but also include aspects in which there are a plurality of members, unless the context clearly indicates otherwise.

[0025] The terms "about" and "approximately," as used herein with respect to a given value, generally mean a deviation from the specified value that is within 30% or within 20% of the specified value. For example, "about" with respect to a dosage or amount is understood by those skilled in the art to provide a pharmacological effect equivalent to that obtained from the specified dosage or amount. In certain embodiments, the terms "about" and "approximately" as used in the context include dosages or amounts that are within 20%, within 15%, within 10%, or within 5% of the specified dosage or amount.

[0026] The present disclosure provides a method of treating a retinal disease in a subject by administering to the subject a pharmaceutically effective amount of a pharmaceutical composition comprising a bisphosphonate-based ASM / FDPS inhibitor, wherein the bisphosphonate-based ASM / FDPS inhibitor is administered at an ultra-low dose. As used herein, an ASM / FDPS inhibitor refers to a compound or agent that inhibits both acid sphingomyelinase (ASM) activity and farnesyl diphosphate synthase (FDPS) activity. In one related embodiment, the present disclosure further provides a pharmaceutical composition comprising a bisphosphonate-based ASM / FDPS inhibitor for use in a method of treating a retinal disease in a subject, wherein the method comprises administering the bisphosphonate-based ASM / FDPS inhibitor at an ultra-low dose. In another aspect, the present disclosure provides a method of making a medicament for treating a retinal disease, the method comprising using a bisphosphonate-based ASM / FDPS inhibitor at an ultra-low dose.

[0027] Ultra-low dose One basic aspect of the present invention is the use of bisphosphonate-based ASM inhibitors, such as zoledronic acid, at substantially lower doses than currently clinically used doses. As used herein, "dose" means the amount of a selected agent delivered in a single administration. For example, an example of a dose of sugar could be "4 grams" or "1 cube of 4 grams of granulated sugar". As used herein, regimen refers to a specific dose of a selected agent delivered over a selected period, which may be a specific number of administrations, a specific frequency, or a specific route of administration. For example, an example of a regimen of sugar could be "8 grams of sugar per day, with a dose of 4 grams administered 2 times, once in the morning and once in the evening, and administered orally".

[0028] In connection with the administration of an ASM / FDPS inhibitor, such as zoledronic acid, for treating retinal diseases, the "low dose" as used herein includes a dose of a bisphosphonate-based ASM / FDPS inhibitor, such as zoledronic acid, that is significantly lower compared to the doses generally used clinically, for example, at least 1 / 5, preferably 1 / 10 or 1 / 100. In some embodiments, an ultra-low dose is at least 1 / 10, preferably 1 / 100, compared to the doses generally used clinically of a bisphosphonate-based ASM / FDPS inhibitor, such as zoledronic acid. For the treatment of osteoporosis and for cancer-related treatments, the standard dose of zoledronic acid is 4 mg or 5 mg, which is delivered in a regimen that includes a single dose of 4 mg or 5 mg of zoledronic acid delivered intravenously once a year. Such administration results in an average of about 40 - 100 μg per kilogram of body weight. As described in the Examples section of this specification, in some embodiments, doses that are at least 1 / 100 lower than the established clinical doses and that can be effectively used to inhibit lipofuscin-mediated lesions in the RPE underlying various retinal pathologies have been discovered by the inventors of the present disclosure.

[0029] In one practice, the method of the present invention includes administering a bisphosphonate ASM / FDPS inhibitor, such as zoledronic acid, to a human subject at a dose of 0.005 to 10 mg or 0.01 to 1 mg, such as a dose of 0.5 mg to 1.0 mg, such as a dose of 0.3 mg to 0.5 mg, such as a dose in the range of 0.01 to 0.30 mg. Exemplary doses include, for example, 0.005 mg, 0.01 mg, 0.025 mg, 0.05 mg, 0.075 mg, 0.10 mg, 0.15 mg, 0.2 mg, 0.25 mg, and 0.3 mg. For smaller human subjects or non-human animal subjects, equivalent doses may be calculated based on known methodologies.

[0030] In one practice, the method of the present invention includes administering a bisphosphonate ASM / FDPS inhibitor, such as zoledronic acid, at a dose of 10 ng to 50 μg per kg of body weight, such as a dose of 100 ng to 20 μg per kg of body weight, such as a dose of 1.0 μg to 10 μg per kg of body weight, such as a dose in the range of 3 to 7 μg per kg of body weight, or a dose of about 5 μg per kg of body weight. Exemplary doses include, for example, 100 ng per kg of body weight, 200 ng per kg of body weight, 300 ng per kg of body weight, 400 ng per kg of body weight, 500 ng per kg of body weight, 600 ng per kg of body weight, 700 ng per kg of body weight, 800 ng per kg of body weight, 900 ng per kg of body weight, 1 μg per kg of body weight, 2 μg per kg of body weight, 3 μg per kg of body weight, 4 μg per kg of body weight, 5 μg per kg of body weight, and 6 μg per kg of body weight, 7 μg per kg of body weight, 8 μg per kg of body weight, 9 μg per kg of body weight, and 10 μg per kg of body weight.

[0031] Typically, the dose range of a bisphosphonate ASM / FDPS inhibitor, such as zoledronic acid, is m 2 in terms of body surface area, 0.001 mg / m 2 to 1.0 mg / m2 in the range of, for example, 0.037 mg / m 2 ~0.37 mg / m 2 in the range of, for example, about 0.05 mg / m 2 , 0.06 mg / m 2 , 0.07 mg / m 2 , 0.08 mg / m 2 , 0.09 mg / m 2 , 0.1.0 mg / m 2 , 0.2 mg / m 2 , 0.3 mg / m 2 , 0.4 mg / m 2 , or 0.5 mg / m 2 is included. The human equivalent dose (HED) of zoledronic acid is in the range of 0.1 μg / kg to 1 μg / kg.

[0032] The doses disclosed above may be administered by various suitable dosing regimens, methods of use, dosage forms, and routes of administration. With respect to frequency, the selected dose may be administered at any selected frequency, for example, once a year, once a month, once a week, once a day, etc. The treatment period may be arbitrary and may be, for example, 1 week, 2 weeks, 3 weeks, 4 weeks, 2 months, 3 months, 6 months, 1 year, or indefinite as needed. Administration may also be carried out when necessary, which is determined by clinical evaluation, or administration may be adjusted with respect to the amount and / or treatment frequency based on clinical evaluation. In addition to or instead of the above, the dose may be increased from at least 1 lower dose to at least 1 higher dose during subsequent administrations. In some examples, in addition to or instead of the above, the dose may be decreased from at least 1 higher dose to at least 1 lower dose during subsequent administrations.

[0033] In addition, administration may be by any selected route of administration. In one practice, administration is systemic administration, which includes, for example, intravenous administration, intraperitoneal administration, subcutaneous administration, or oral administration. In such a practice, the bisphosphonate ASM / FDPS inhibitor administered, such as zoledronic acid, is typically administered at a relatively high dose compared to local administration to the eye, for the purpose of achieving a therapeutically effective concentration in the RPE, which is, for example, a dose of 0.01 to 1 mg per human subject, a dose of 1 to 30 μg per kg of body weight, and a dose of 0.037 to 1.0 mg per m 2 of body surface area, etc.

[0034] In another practice, administration includes ocular administration, i.e., administration to the ocular compartment. For example, a selected bisphosphonate ASM / FDPS inhibitor, such as zoledronic acid, may be administered by intravitreal delivery, which is, for example, by methods such as suprachoroidal injection, subconjunctival injection, intravitreal injection, subretinal injection, or sub-Tenon's injection. In such a practice, the bisphosphonate ASM / FDPS inhibitor administered can avoid the blood-brain barrier (BBB), as well as systemic dilution and clearance, and can achieve a therapeutically effective concentration in the RPE at a relatively low dose compared to the dose that can be administered systemically. For example, in some embodiments, the dose delivered by intravitreal delivery may include a dose of 0.001 to 0.1 mg per human subject, a dose of 0.05 to 2 μg per kg of body weight, and a dose of 0.001 to 0.1 mg per m 2 of body surface area.

[0035] In one related practice, the treatment method using intravitreal administration may further include an anti-inflammatory regimen during the period before and after injection. The anti-inflammatory regimen may include steroids and non-steroidal anti-inflammatory drugs, alone or in combination. The anti-inflammatory drugs may be administered, alone or in combination, via the oral route, the intravitreal route, and / or the topical route to the eye.

[0036] In one related practice, administration to the eye is topical administration to the eye, which is effected, for example, by use of a gel, ophthalmic ointment, or eye drops applied to the outer surface of the eye. Exemplary dosage forms include eye drops or gels, which contain, for example, a solution, suspension, emulsion, or other preparation of a selected bisphosphonate ASM / FDPS inhibitor, such as zoledronic acid, in a carrier. In such a practice, delivery is expected to be more efficient than systemic delivery but less efficient than intraocular delivery, as the agent needs to pass through the vitreous membrane and the intraocular fluid, as well as the photoreceptor cells, to reach the RPE. In some embodiments, the dosage topically applied to the eye may include a dosage of 0.01 to 0.5 mg per human subject, a dosage of 0.5 to 8 μg per kg body weight, and a dosage of 0.0185 to 0.3 mg per m 2 2 of body surface area.

[0037] In some embodiments, the dosage for humans is based on the dosage for animals, for example, based on the dosage scaling conversions often used in the art (see, for example: Nair & Jacob, J. Basic and Clin. Pharmac 2016; 7:27-31). Thus, for example, to convert the animal dosage in mg / kg units to the human equivalent dosage in mg / kg units, the animal dosage would be divided by 12.3.

[0038] In some embodiments, zoledronic acid is formulated as an ophthalmic solution or suspension for administration as eye drops, which can be formulated, for example, as a solution, suspension, liposomal formulation, mucoadhesive polymer, micelle, nanoparticles, etc. (see, for example: Kim et al, Drug Delivery and Translational Research 12:826-837, 2022). In some embodiments, the ophthalmic solution or suspension provides a dose of zoledronic acid per eye of about 10 ng / day to about 250 μg / day. In some embodiments, the ophthalmic solution or suspension provides a dose of zoledronic acid per eye of about 10 ng / day to about 100 μg / day. In some embodiments, the eye drops are formulated to provide a dose of zoledronic acid per eye of about 50 ng / day to about 50 μg / day. In some embodiments, the eye drops are formulated to provide a dose of zoledronic acid per day of about 100 ng / day to about 50 μg / day. In some embodiments, the eye drops are formulated to provide a dose of zoledronic acid per eye of about 50 ng / day, or about 100 ng / day, up to about 10 μg / day. In some embodiments, the eye drops are formulated to provide a dose of zoledronic acid per eye of at least about 50 ng / day, at least 100 ng / day, at least 200 ng / day, at least 250 ng / day, at least 300 ng / day, at least 350 ng / day, at least 400 ng / day, at least 500 ng / day, at least 600 ng / day, at least 700 ng / day, at least 800 ng / day, at least 900 ng / day, at least μg / day, at least 1.5 μg / day, at least 2 μg / day, at least 2.5 μg / day, at least 3 μg / day, at least 4 μg / day, at least 5 μg / day, at least 10 μg / day, at least 15 μg / day, at least 20 μg / day, or at least 25 μg / day, but up to about 50 μg / day.In some embodiments, the eye drops are formulated to provide zoledronic acid at a constant intraocular concentration of at least 100 pM, 1 nM, 10 nM, 100 nM, 200 nM, 500 nM, or 1 μM. In some embodiments, the ophthalmic solution is administered once daily to provide a specified daily dose. In some embodiments, the ophthalmic solution is administered multiple times a day, such as twice or three times a day, to provide a specified daily dose. In some embodiments, the ophthalmic solution may be administered at least daily, twice a week, three times a week, four times a week, or more often. In some embodiments, zoledronic acid may be administered once a week or once every two weeks. In some embodiments, zoledronic acid may be administered once a month or twice a month.

[0039] In some embodiments, zoledronic acid is delivered to the eye using an intravitreal implant. In such embodiments, zoledronic acid may be incorporated into the implant to provide release of zoledronic acid over a period of up to several years, for example, up to 2 or 3 years. In some embodiments, the implant may be loaded with an amount of zoledronic acid in the range of about 0.001 to about 0.3 mg. In some embodiments, zoledronic acid may be incorporated into the implant to provide release of zoledronic acid over a period shorter than a period of several weeks to several months, for example, over a period shorter than 2 - 4 weeks, or 4 - 6 weeks, or 6 - 8 weeks, or 3 - 6 months, or 6 - 12 months, or 12 - 18 months. In some embodiments, the implant may be a biodegradable polymer. In some embodiments, the implant is loaded with an amount of zoledronic acid to provide release of a dose in the range of about 0.001 μg / day to about 2.5 μg / day, or release of a dose in the range of about 0.005 μg / day to about 2.0 μg / day. In some embodiments, the implant is loaded with an amount of zoledronic acid to provide a constant intraocular concentration of zoledronic acid of at least 100 pM, 1 nM, 10 nM, 100 nM, 200 nM, 500 nM, or 1 μM. In some embodiments, the implant is loaded with zoledronic acid to provide a dose in the range of about 10 ng / day to about 250 μg / day. In some embodiments, the implant is loaded to provide a dose of zoledronic acid in the range of about 50 ng / day to about 100 μg / day. In some embodiments, the implant is loaded to provide a dose of zoledronic acid in the range of about 100 ng / day to about 50 μg / day. In some embodiments, the implant is loaded to provide a dose of zoledronic acid in the range of about 100 ng / day to about 10 μg / day.In some embodiments, the implant is loaded to provide a dose of zoledronic acid that is at least about 50 ng / day, at least 100 ng / day, at least 200 ng / day, at least 250 ng / day, at least 300 ng / day, at least 350 ng / day, at least 400 ng / day, at least 500 ng / day, at least 600 ng / day, at least 700 ng / day, at least 800 ng / day, at least 900 ng / day, at least 1 μg / day, at least 1.5 μg / day, at least 2 μg / day, at least 2.5 μg / day, at least 3 μg / day, at least 4 μg / day, at least 5 μg / day, at least 10 μg / day, at least 15 μg / day, at least 20 μg / day, or at least 25 μg / day, but is about 50 μg / day or less or 100 μg / day or less. In some embodiments, the sustained release implant is a liquid that is immiscible or partially immiscible and includes, but is not limited to, benzyl benzoate and / or silicone oil. Such an embodiment can include a suspension, emulsion, or solution of zoledronic acid.

[0040] In some embodiments, suprachoroidal injection is used to administer zoledronic acid. In some embodiments, a pharmaceutical composition of zoledronic acid in which zoledronic acid is conjugated to a dendrimer (see, for example, Pitha et al, Biomacromolecules 24:1355-1365, 2023) or in which zoledronic acid is provided as a nanoparticle formulation (see, for example, Pitha et al, Biomacromolecules 24:1355-1365, 2023; Laradji et al, Polymers 13(19), 3324, 2021) is utilized. In some embodiments, an injectable solution or suspension is formulated to provide zoledronic acid at a concentration of 1 μg / ml to 10 mg / ml and is delivered by suprachoroidal injection or intravitreal injection. In some embodiments, an injectable solution or suspension containing zoledronic acid, such as zoledronic acid conjugated to a dendrimer or zoledronic acid in a nanoparticle formulation, is formulated to deliver zoledronic acid in an amount of about 0.1 to 1 mg / eye. In some embodiments, the amount of zoledronic acid delivered by injection into the eye, such as by suprachoroidal injection or intravitreal injection, is a dosage of about 10 ng / day to about 50 μg / day. In some embodiments, the amount of zoledronic acid delivered by injection into the eye, such as by suprachoroidal injection or intravitreal injection, is a dosage of about 10 ng / day to about 50 μg / day. In some embodiments, the dosage of injectable zoledronic acid is about 100 ng / day to about 50 ug / day. In some embodiments, the dosage of injectable zoledronic acid is about 100 ng / day to about 10 μg / day.In some embodiments, the injectable is loaded to provide zoledronic acid at a dosage of at least about 50 ng / day, at least 100 ng / day, at least 200 ng / day, at least 250 ng / day, at least 300 ng / day, at least 350 ng / day, at least 400 ng / day, at least 500 ng / day, at least 600 ng / day, at least 700 ng / day, at least 800 ng / day, at least 900 ng / day, at least 1 μg / day, at least 1.5 μg / day, at least 2 μg / day, at least 2.5 μg / day, at least 3 μg / day, at least 4 μg / day, at least 5 μg / day, at least 10 μg / day, at least 15 μg / day, at least 20 μg / day, or at least 25 μg / day and less than 100 μg / day, preferably less than about 50 μg / day. In some embodiments, an injectable composition, such as a composition for suprachoroidal or intravitreal injection, is formulated to provide a zoledronic acid intraocular concentration of at least 100 pM, 1 nM, 10 nM, 100 nM, 200 nM, 500 nM, or 1 μM.

[0041] In some embodiments, the ophthalmic or systemic composition may be administered using a prefilled syringe.

[0042] Bisphosphonate ASM / FDPS inhibitor The methods of the invention include the use of a bisphosphonate ASM / FDPS inhibitor for treating a selected retinal disorder. In one basic embodiment, the bisphosphonate ASM / FDPS inhibitor is zoledronic acid or a pharmaceutically effective variant thereof. As used herein, zoledronic acid includes any form of therapeutically active zoledronic acid, including Structure 1, its pharmaceutically effective salts, its acid anhydride forms, its hygroscopic forms, and its hydrate forms, its lipophilic derivatives, and its chemically related derivatives.

[0043] In one aspect, zoledronic acid is provided as a salt, which can be, for example, an arginine salt, a calcium salt, a chromium salt, a citrulline salt, a cobalt salt, a copper salt, a creatine salt, a glutamine salt, a histidine salt, an iron salt, an isoleucine salt, a leucine salt, a lithium salt, a lysine salt, a magnesium salt, a manganese salt, a molybdenum salt, an ornithine salt, a potassium salt, a selenium salt, a sodium salt, a zinc salt, or any combination of the above.

[0044] In one aspect, the bisphosphonate ASM / FDPS inhibitors include the derivatives of zoledronic acid disclosed below: U.S. Patent No. 4,939,130, Substituted alkanediphosphonic acids and pharmaceutical use, by Jaeggi and Wilder. In one aspect, the bisphosphonate ASM / FDPS inhibitors include the derivatives of zoledronic acid disclosed below: International Publication No. 2012071517 of PCT International Patent Application, Novel Crystalline Forms, by Hanna et al. In one aspect, the bisphosphonate ASM inhibitors include the derivatives of zoledronic acid disclosed below: U.S. Patent Application Publication No. 20100056481, Crystalline forms of zoledronic acid, by Glausch et al. In another aspect, the bisphosphonate ASM / FDPS inhibitors include derivatives of zoledronic acid conjugated with deoxycholic acid with lysine linked to increase oral absorption as described below: Jeon et al., 2016. In another aspect, the bisphosphonate ASM / FDPS inhibitors include the compositions disclosed below: U.S. Patent No. 9,682,091, named Oral Forms of a Phosphonic Acid Derivative.

[0045] In one practice, bisphosphonate ASM / FDPS inhibitors include zoledronic acid in a form configured for oral delivery. Oral delivery conveniently avoids the inconvenience and risk of side effects of eye injections. For example, in one aspect, bisphosphonate ASM / FDPS inhibitors include forms of zoledronic acid having oral bioavailability as disclosed below: US Patent Application Publication No. 20140051669 by Tabuteau et al., Compositions of Zoledronic Acid or Related Compounds for Treating Disease; or International Publication No. 2013015599 of PCT International Patent Application by Kim et al., Pharmaceutical Composition for Oral Administration Comprising Bisphosphonic Acid or its Salt.

[0046] Adiporone In a further aspect of the present disclosure, adiporone (PubChem CID 16307093) is used to treat selected retinal pathologies. Adiporone is a synthetic small molecule agonist of adiponectin receptor 1 (AdipoR1) that is selective and orally active, and is a synthetic adiponectin receptor agonist of adiponectin receptor 2 (AdipoR2). In some aspects, adiporone is administered in the amounts described herein with respect to the administration of bisphosphonate ASM / FDPS inhibitors. In some aspects, adiporone is utilized at a dose of 1 / 10 to 1 / 1000 compared to the doses used for other indications.

[0047] Treatment of Retinal Pathologies The method of the present invention is directed to the treatment of retinal pathologies in a subject in need of treatment of retinal pathologies by administering a low dose of a bisphosphonate ASM / FDPS inhibitor.

[0048] With respect to the subject, the subject may be any animal subject in need of treatment for a retinal disorder. The subject may be of any animal species. In one basic aspect, the subject is a human, such as a human patient. In other practices, the subject is a non-human animal, such as a veterinary subject, a pet, a livestock animal, or a laboratory animal. Exemplary non-human animals include mice, rats, pigs, horses, cows, dogs, cats, non-human primates, and others.

[0049] The subject may be a subject suffering from or diagnosed with a selected retinal disorder, or a subject suspected of having a selected retinal disorder. In one aspect, the subject is a subject at risk of developing a selected retinal disorder. In one aspect, the subject is an elderly subject, such as a human subject who is at least 50 years old, at least 55 years old, at least 60 years old, or at least 65 years old. In one aspect, the subject is a subject having one or more genetic markers indicative of a risk of a retinal disorder, or a subject having a family history of a retinal disorder with a risk of the retinal disorder being inherited. In one practice, the subject is at risk of, suspected of having, or suffering from dry age-related macular degeneration.

[0050] In some examples, the subject is a human child, teenager, or young adult, such as a human 25 years of age or younger, who is at risk of, suspected of having, or diagnosed with a retinal disease. In one aspect, the subject is at risk of, suspected of having, or suffering from Stargardt macular dystrophy, which is also referred to herein as Stargardt's disease. In some aspects, the subject is an adult over 25 years of age who is at risk of, suspected of having, or has Stargardt macular dystrophy. In some aspects, the subject has autosomal dominant Stargardt's disease. In some examples, the subject has autosomal recessive Stargardt's disease.

[0051] Regarding retinal pathologies treatable by the method of the present invention, the retinal pathology may be any pathology of the retina in which RPE dysfunction, microglial activation, or photoreceptor damage is known or suspected, including pathologies characterized by or associated with any of the following: lipofuscin accumulation in RPE cells, cholesterol accumulation in RPE cells, abnormal activation of ASM or FDPS in RPE cells, abnormal ceramide production in RPE cells, abnormal prenylation of Rab GTPase in the RPE, abnormal microtubule acetylation in RPE cells, autophagy deficiency in RPE cells, drusen accumulation or similar deposits above and below the RPE, complement-mediated mitochondrial damage in the RPE, presence of subretinal microglia, or loss or dysfunction of photoreceptors. As described above, in one basic practice, the pathology is dry AMD. In other practices, the retinal pathology is Stargardt macular dystrophy, including, for example: autosomal dominant or autosomal recessive Stargardt disease; Doyne honeycomb dystrophy; acid ceramidase diseases such as Faber disease; diseases with cholesterol accumulation, ceramide accumulation, and autophagy defects such as Niemann-Pick disease type C; and diseases such as Batten disease (neuronal ceroid lipofuscinosis), Best vitelliform macular dystrophy; retinitis pigmentosa; and Bietti crystalline dystrophy.

[0052] The methods of the present invention include the treatment of selected retinal pathologies. As used herein, "treatment" includes achieving various therapeutic effects and treatment outcomes related to the selected retinal pathology, and includes, for example: ameliorating symptoms associated with the selected retinal pathology; delaying the progression of the selected retinal pathology; preventing further damage to the RPE due to the selected retinal pathology; improving the function of the RPE; maintaining, improving, or restoring the integrity of photoreceptor cells; maintaining, improving, or restoring vision; or any other reduction in the pathological condition associated with the selected retinal pathology. As used herein, treatment further includes preventing the selected retinal pathology. For example, prophylactic treatment may include achieving any of the following: preventing or delaying the onset of the selected retinal pathology in at-risk subjects; maintaining normal vision or RPE function; or preventing the onset of the selected retinal pathology. As used herein, treatment further includes: preventing the progression of geographic atrophy, delaying the progression of geographic atrophy, treating angiogenesis prevention of the progression of neovascular AMD, reducing the number and / or frequency of other treatments for AMD (such as anti-VEGF injections, complement inhibitors, etc.), changes in the total area of geographic atrophy (GA) based on fundus autofluorescence, reduction in photoreceptor cell loss (such as as measured by the area where the ellipsoid zone is unclear), changes in reading speed under standard and low light conditions, changes in contrast sensitivity, changes in retinal sensitivity in visual field measurements, changes in patient-reported outcomes, changes in dark adaptation, changes in drusen amount, changes in pigmented spot formation, in well-defined macular atrophy, in target macular disease, or in fundus spots, changes in electroretinogram (ERG), and / or structural changes as measured by optical coherence tomography (OCT).

[0053] Treatment further includes any inhibition of the pathological processes underlying the selected retinal pathologies. Exemplary treatment effects include, for example: reducing lipofuscin accumulation in the RPE, reducing cholesterol accumulation in RPE cells, reducing ASM and / or FDPS activity in RPE cells, reducing abnormal ceramide production in RPE cells, reducing abnormal prenylation of Rab, reducing abnormal microtubule acetylation in RPE cells, improving or restoring autophagic capacity, autophagic activity, and autophagic flux in RPE cells, reducing drusen accumulation or the formation of similar deposits, reducing complement-mediated mitochondrial damage in the RPE, preventing or rescuing the migration of microglia into the subretinal space, or preventing or rescuing photoreceptor loss and dysfunction. As demonstrated in Example 3 herein, inhibition of ASM and FDPS with bisphosphonates using zoledronic acid effectively reduces the accumulation of lipofuscin, cholesterol, and ceramide, which is significantly different from previously investigated agents such as desipramine, which only reduces ceramide. Thus, in one aspect, the scope of the present invention further includes methods of reducing lipofuscin accumulation, cholesterol accumulation, and / or ceramide accumulation by administering zoledronic acid or a derivative thereof, for example, at ultra-low doses.

[0054] The treatment of the present invention includes administering to a subject one or more bisphosphonate ASM / FDPS inhibitors in a therapeutically effective amount. In one criterion, a therapeutically effective amount is an amount of a bisphosphonate ASM / FDPS inhibitor sufficient to induce a measurable therapeutic effect. The therapeutic effect may be the achievement of a selected physiological outcome or physiological state, which may be, for example: a reduction in the accumulation of lipofuscin, cholesterol, or ceramide; a reduction in ASM activity and FDP activity; an improvement in autophagic flux in the RPE; a reduction in drusen formation by RPE cells; a reduction in complement-mediated mitochondrial damage in RPE cells; an improvement in the integrity and function of the RPE; an improvement in the integrity and function of photoreceptor cells; a reduction in microglial activation; and an improvement in vision. As used herein, "subject" refers to any mammal, including non-human primates, mice, rats, rabbits, pigs, horses, cows, goats, sheep, dogs, cats, etc. In a preferred embodiment, the subject is a human.

[0055] Pharmaceutical composition The methods and compositions of the present invention include the administration of one or more bisphosphonate ASM / FDPS inhibitors, such as zoledronic acid. One or more bisphosphonate ASM / FDPS inhibitors may be formulated as what is referred to as a "pharmaceutical composition." As used herein, a pharmaceutical composition includes one or more bisphosphonate ASM inhibitors and may further include various additional compositions, including excipients, carriers, diluents, release formulations, drug delivery vehicles or drug targeting vehicles, and additional active therapeutic agents. The pharmaceutical compositions of the present invention may be formulated to be compatible with a selected route of administration, such as oral delivery, delivery by injection, or delivery to the eye.

[0056] The pharmaceutical composition of the present invention may include one or more bisphosphonate ASM / FDPS inhibitors, such as zoledronic acid, in combination with a drug delivery composition. The drug delivery composition includes any moiety, any substance, or any other composition that promotes the delivery of the ASM / FDPS inhibitor, such as zoledronic acid, to the RPE. In some practices, the drug delivery composition facilitates targeting of the ASM / FDPS inhibitor, such as zoledronic acid, to the CNS, the PNS, or other selected target compartments of the nervous system. In some practices, the delivery composition includes a composition that facilitates passage through the blood-brain barrier. In some practices, the delivery composition includes a composition that facilitates transport across the blood-retinal barrier (BRB).

[0057] The pharmaceutical composition may include any form of combination, including the following: functionalization of a bisphosphonate ASM / FDPS inhibitor (such as zoledronic acid) with a delivery composition, conjugation of a bisphosphonate ASM / FDPS inhibitor with a delivery composition; mixing of a bisphosphonate ASM / FDPS inhibitor with a delivery composition; encapsulation or injection of a bisphosphonate ASM / FDPS inhibitor into a delivery composition, or any other combination.

[0058] In one practice, the delivery composition includes a bisphosphonate-based ASM / FDPS inhibitor, such as zoledronic acid, and a substance that facilitates passage through the BBB or BRB. For example, the delivery composition may include a ligand that promotes transcytosis through the endothelial cells of the brain to pass through the BBB from the apical side to the basolateral side, which may be, for example, an anti-transferrin receptor antibody or an antigen-binding fragment thereof, such as the OX26 antibody, a polypeptide such as Angiopep2, the ApoE protein and its mimetics; diphtheria toxin, and surfactants, etc. Further targeting moieties include BBB-permeating peptides, which may be, for example, those described below: Van Dorpe et al., "Brainpeps: The blood-brain barrier peptide database", Brain Structure and Function, 2012, 217(3), 687-718. In addition, the delivery composition may include a hypertonic or hyperosmotic formulation that promotes BBB transport or BRB transport.

[0059] In one practice, to facilitate passage across the BBB and / or to facilitate delivery to target RPE cells, the delivery composition comprises a carrier, and a bisphosphonate-based ASM / FDPS inhibitor, such as zoledronic acid, is conjugated to the carrier, encapsulated within the carrier, or combined with the carrier. Exemplary carriers include the following: liposomes; extracellular vesicles or synthetic mimics thereof, such as exosomes; red blood cells modified with a bisphosphonate-based ASM / FDPS inhibitor; microspheres, such as poly(lactic-co-glycolic acid) (PLGA) microspheres; and other drug delivery nanoparticles, such as PLGA-PEG nanoparticles, alginate nanoparticles or chitosan nanoparticles, silica nanoparticles, and iron oxide nanoparticles. The carrier molecule or carrier composition may be further functionalized with a ligand that facilitates passage across the BBB or BRB, and the ligand may be, for example, an anti-tfR antibody, a polypeptide such as Angiopep2, an ApoE protein and mimics thereof; diphtheria toxin, and surfactants, as described above.

[0060] In some embodiments, the bisphosphonate-based ASM / FDPS inhibitor, such as zoledronic acid, is administered as a drug-antibody conjugate, which comprises, for example, an antibody that targets a ligand present on the RPE, or an antigen-binding fragment thereof.

[0061] In one aspect, the delivery composition constitutes or is incorporated into an implant, which is, for example, a drug-eluting implant that is placed into a target tissue, such as into the eye. Exemplary implants include, for example: implants made of biodegradable materials, such as PLGA, polymeric drug-eluting wafers, polymeric drug-eluting rods, injectable hydrogels, implantable hydrogel scaffolds, hydrophilic microsphere-based systems, cyclodextrin-based systems, polymer micelle-based systems, and other drug-eluting implants known in the art. Exemplary implants for delivering an agent to the RPE include those described below: International Publication No. WO 2012177968 by Tao et al., "A scaffold for subretinal cell transplantation and drug delivery". Exemplary polymers and implants for drug delivery to the eye are further described by: Allyn et al, Frontiers in Medicine, 8: Article 787644, January 2022; Cao et al, Drug Discovery Today 24(8), 1694 - 1700 2019).

[0062] In one practice, the delivery composition constitutes an intravitreal implant containing zoledronic acid loaded with an amount of zoledronic acid in the range of 0.001 to 0.3 mg. In some aspects, the implant is loaded with 0.005 to 2.5 mg of zoledronic acid.

[0063] In yet another aspect, the delivery composition includes an ophthalmic topical preparation containing zoledronic acid at 0.001 to 0.05 mg / dose. In some aspects, the preparation contains zoledronic acid at 0.005 to 0.05 mg / dose. In some aspects, the ophthalmic topical preparation is a solution or suspension that is applied as an eye drop.

[0064] In yet another aspect, the delivery composition comprises an injectable ophthalmic preparation, which is, for example, an ophthalmic preparation suitable for suprachoroidal or intravitreal injection, containing zoledronic acid at a concentration of 1 μg / ml to 10 mg / ml. In some aspects, the injectable ophthalmic preparation contains zoledronic acid at a concentration of 10 μg / ml to 1 mg / ml. In some aspects, the injectable ophthalmic preparation contains zoledronic acid conjugated to a dendrimer or formulated as nanoparticles.

[0065] It is understood that combination administration of a bisphosphonate ASM / FDPS inhibitor, such as zoledronic acid, with one or more additional active agents for treating retinopathy is further encompassed by the methods of the present invention. Additional treatments, such as additional treatments for macular degeneration or Stargardt disease, include but are not limited to: anti-VEGF treatments (including but not limited to rolucizumab, aflibercept, ranibizumab, pegaptanib sodium, faricimab suba, and bevacizumab), complement inhibitors (including but not limited to pegcetacoplan and abicipar pegol), MCO 010, deuterated retinol, emixustat, MA09 hRPE, STG-001, chinrarenbant, QR1011, and REV 0100. As used herein, combination administration can encompass any combination of administration of a bisphosphonate ASM / FDPS inhibitor and one or more additional treatments for retinopathy. For example, the timing of administration of the ASM / FDPS inhibitory treatment and one or more additional treatments can be determined by one of ordinary skill in the art. In various practices, the ASM / FDPS inhibitory treatment and the additional treatment are administered either concurrently, sequentially, or alternately. In one aspect, the first treatment and the second treatment are applied concurrently, i.e., simultaneously or with overlapping timing. In one aspect, the first treatment and the second treatment are administered as a pharmaceutical composition comprising a combination product, the combination product comprising a bisphosphonate ASM / FDPS inhibitor and an additional agent, which is administered, for example, in a single dosage form. In one aspect, the one or more additional treatments include, for example, administration of an adiponectin 1 receptor agonist as described below: Kim et al, 2022. Adiponectin receptor agonists promote improvement of cardiac lipotoxicity by enhancing the metabolism of ceramide in type 2 diabetic mice. Cell Death & Disease 13: 282. In one aspect, the adiponectin 1 receptor agonist is adiporone.In one aspect, the pharmaceutical composition of the present invention comprises a combination product, and the combination product comprises a bisphosphonate ASM / FDPS inhibitor combined with one or more adiponectin 1 receptor agonists, such as adiporone.

[0066] The pharmaceutical composition of the present invention can be formulated into various dosage forms. Exemplary dosage forms include: liquid solutions; sachets, capsules, or tablets, each containing a predetermined amount of the active ingredient as a solid or granule; suspensions in liquids; emulsions; aqueous and non-aqueous solutions; isotonic sterile injection solutions; compositions stored in a freeze-dried and lyophilized state; and other dosage forms known in the art. In one aspect, the dosage form is formulated for topical application to the eye and includes eye drops, ophthalmic ointments, or gels.

[0067] Exemplary aspects include, but are not limited to: Aspect 1: A method for treating a retinal disease in a subject in need of treatment for a retinal disease, the method comprising administering to the subject a pharmaceutically effective amount of a pharmaceutical composition comprising a bisphosphonate ASM / FDPS inhibitor, wherein the bisphosphonate ASM / FDPS inhibitor is administered at an ultra-low dose. Aspect 2: The method according to aspect 1, wherein the retinal disease is a condition mediated by the accumulation of lipofuscin in RPE cells. Aspect 3: The method according to aspect 1, wherein the retinal disease is dry age-related macular degeneration. Aspect 4: The method according to aspect 1, wherein the retinal disease is selected from the group consisting of Stargardt macular dystrophy, neuronal ceroid lipofuscinosis, Batten disease, Best vitelliform macular dystrophy, Niemann-Pick disease type C, Doin honeycomb dystrophy, Faber disease, and Best vitelliform macular dystrophy. Aspect 5: The method according to aspect 1, wherein the bisphosphonate ASM / FDPS inhibitor is zoledronic acid or a derivative thereof. Aspect 6: The method according to aspect 1, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.005 to 2.0 mg. Aspect 7: The method according to aspect 6, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.1 to 1.0 mg. Aspect 8: The method according to aspect 7, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.3 to 0.5 mg. Aspect 9: The method according to aspect 6, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose selected from the group consisting of 0.005 mg, 0.006 mg, 0.007 mg, 0.008 mg, 0.009 mg, 0.01 mg, 0.02 mg, 0.03 mg, 0.04 mg, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.10 mg, 0.20 mg, 0.30 mg, 0.40 mg, 0.50 mg, 0.60 mg, 0.70 mg, 0.80 mg, 0.90 mg, and 1.0 mg. Aspect 10: The method according to aspect 1, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 100 ng to 10 μg per kg of body weight. Aspect 11: The method according to aspect 10, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 1.0 μg to 7.0 μg per kg of body weight. Aspect 12: The method according to aspect 11, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of about 5.0 μg per kg of body weight. Aspect 13: The method according to aspect 10, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose selected from the group consisting of 0.1 μg, 0.2 μg, 0.3 μg, 0.5 μg, 1.0 μg, 2.0 μg, 3.0 μg, 4.0 μg, 5.0 μg, 6.0 μg, 7.0 μg, 8.0 μg, 9.0 μg, and 10.0 μg per kg of body weight. Aspect 14: The method according to any one of aspects 1 to 13, wherein the pharmaceutical composition is administered at a frequency selected from the group consisting of once a year, once a month, twice a month, once a week, twice a week, every other day, once a day, twice a day, and three times a day. Aspect 15: The method according to any one of Aspects 1 to 4, wherein the administration is by a route including any one of systemic delivery; local delivery; intravenous delivery; intramuscular delivery; intraperitoneal delivery; topical delivery; subcutaneous delivery; intravitreal delivery; suprachoroidal delivery, and topical delivery to the eye. Aspect 16: The method according to any one of Aspects 1 to 15, wherein the pharmaceutical composition includes one or more bisphosphonate-based ASM / FDPS inhibitors, and includes any one of an excipient, a carrier, a diluent, a release formulation, a drug delivery vehicle or a drug targeting vehicle, and an additional active therapeutic agent. Aspect 17: The method according to Aspect 16, wherein the pharmaceutical composition includes an adiponectin 1 receptor agonist. Aspect 18: The method according to Aspect 17, wherein the adiponectin 1 receptor agonist includes adiporone. Aspect 19: The method according to any one of Aspects 1 to 18, wherein the pharmaceutical composition includes any one of a solution; a suspension in a liquid; an emulsion; an isotonic sterile injection solution; a composition in a lyophilized state; an eye drop solution; or an ophthalmic ointment or gel. Aspect 20: The method according to any one of Aspects 1 to 19, wherein the pharmaceutical composition constitutes or is incorporated into an implant; a drug eluting device, a drug eluting structure, or a drug eluting substance; a polymeric drug eluting wafer; an injectable hydrogel; and an implantable hydrogel scaffold. Aspect 21: The method according to Aspect 1, wherein the bisphosphonate-based ASM is co-administered with an adiponectin 1 receptor agonist. Aspect 22: The method according to Aspect 21, wherein the adiponectin 1 receptor agonist is adiporone.

Example

[0068] Example 1 In Vitro Screening of ASM Inhibitors A test was initiated to evaluate ASM inhibitors for treating retinal pathologies associated with the pathological accumulation of lipofuscin.

[0069] The primary RPE cell cultures were established using cells from porcine eyes. The cultured RPE cells were treated with lipofuscin bisretinoid A2E, which has been previously shown to activate ASM. The cultures were subsequently treated with various ASM inhibitors, and measurements of lipofuscin-mediated damage were evaluated. The ASM inhibitors were desipramine, amitriptyline, fluoxetine, and zoledronic acid, which were each applied at 10 μM and added to the cultures 48 hours after A2E treatment.

[0070] Acetylated tubulin in cultured RPE cells was evaluated by immunostaining using specific antibodies. For example, as previously described in FIG. 1 and Toops et al., 2015, the accumulation of lipofuscin traps cholesterol within RPE cells, causing abnormal activation of ASM. ASM activity causes the accumulation of ceramide, which in turn promotes the acetylation of microtubules. This acetylation of microtubules is unstable and inhibits autophagy in RPE. One important function of RPE is to degrade cell debris by autophagy and to remove the tips of the outer segments of photoreceptor cells that overlay it. The acetylation of microtubules interferes with the normal movement of autophagosomes and also interferes with the interaction between autophagosomes and lysosomes, which is essential for autophagy activity in RPE. This results in the accumulation of intracellular debris within RPE, which can cause metabolic stress in RPE and initiate drusen formation. The acetylation of microtubules also renders RPE sensitive to complement-mediated mitochondrial damage, thereby causing the formation of drusen, which are aggregates of damaged material. Thus, the abundance of acetylated microtubules in RPE provides a measure of lipofuscin-induced damage and a measure of the activation of the pathological cascade described above.

[0071] Primary RPE cultures treated with A2E had a significant increase in acetylated tubulin compared to RPE cells not exposed to A2E, indicating autophagic incompetence and resulting damage (Figure 2B). Furthermore, application of an ASM inhibitor following A2E treatment significantly reversed this A2E-induced effect. Among the ASM inhibitors tested, zoledronic acid produced the most potent decrease in the abundance of acetylated tubulin.

[0072] RPE cells treated with A2E and subsequently applied with an ASM inhibitor were further evaluated for autophagic flux by live cell imaging. An autophagy reporter (tfLC3), which contains microtubule-associated protein 1A / 1B light chain 3 (LC3), a target of autophagy, tandemly fused with red fluorescent protein (mRFP) and enhanced green fluorescent protein (EGFP), was applied to cultured RPE cells. When tfLC3 is incorporated by autophagosomes and delivered to lysosomes, the EGFP signal of the reporter quenches under acidity, thus by simultaneously monitoring the mRFP signal and the EGFP signal, a means is provided to observe and quantify autophagosome in RPE cells, where a decrease in the ratio of the EGFP signal to the mRFP signal indicates autophagic activity.

[0073] As shown in Figure 2D, the EGFP:mRFP ratio was significantly increased in RPE cells treated with A2E compared to control cells not exposed to A2E, indicating autophagic incompetence. Application of an ASM inhibitor restored autophagy in A2E-treated cells. Zoledronic acid most effectively promoted autophagy.

[0074] Example 2 Development of Ultra-Low Dose Zoledronic Acid Next, based on the results described in Example 1, the in vivo effect of zoledronic acid on lipofuscin-induced RPE damage was evaluated. Zoledronic acid is Abca4- / - It was administered to mice. Autosomal recessive Stargardt disease is a pathological condition that includes macular degeneration characterized by the accumulation of lipofuscin in the RPE. This type of Stargardt disease is caused by mutations in the ABCA4 gene. Thus, Abca4 - / - mice provide a model for studying dry AMD, Stargardt disease, and other pathological conditions in which pathological accumulation of lipofuscin occurs.

[0075] In the first in vivo study, single intraperitoneal injections of zoledronic acid were administered to mice. The selected dose was 1 mg per kilogram of body weight, or approximately 28 μg per mouse. This dose was selected based on the approved zoledronic acid doses used to treat osteoporosis and cancer in humans. Although negative side effects are known when using zoledronic acid, this drug is generally considered to be highly tolerable. However, significant adverse effects were observed in mice when zoledronic acid was administered at a dose of 1 mg per kilogram of body weight. The side effects included osteonecrosis of the jaw, feeding disorders, and other symptoms such as weakness and lethargy.

[0076] Based on the above results, the effect of administering much lower doses of zoledronic acid to Abca4 - / - mice was evaluated. When zoledronic acid was administered to Abca4 - / - mice as a single intraperitoneal injection at a dose of 5 μg per kilogram of body weight, this was 1 / 100 compared to the dose prescribed to treat osteoporosis and cancer.

[0077] Accumulation of ceramide 16-month-old wild-type mice and Abca4 - / -Mice were administered vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks by intraperitoneal injection (i.p.). At the end of the study, eyes were enucleated and the cornea, lens, vitreous, and retina were removed. Four to six incisions were made to flatten the eyecup and an RPE flat mount was prepared, which was fixed in 4% paraformaldehyde. Ceramide levels were measured by immunostaining using a specific anti-ceramide antibody (Enzo), which has been widely validated by our group and other groups (Toops et al., 2015; Tan et al., 2016; Kaur et al., 2018; La Cunza et al., 2021). Images were captured on a Nikon spinning disk confocal microscope and the fluorescence intensity of ceramide per cell was quantified using Imaris (Bitplane). The results shown in Figure 3 demonstrate that administration of low-dose zoledronic acid decreased ceramide accumulation.

[0078] Cholesterol accumulation 12-month-old wild-type mice and Abca4 - / - Mice were administered vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks by intraperitoneal injection (i.p.). At the end of the study, the cornea, lens, and vitreous were removed and the eyecup was embedded for cryosectioning. 10-μm-thick cryosections of the retina were stained with filipin (Sigma), which specifically binds to free cholesterol (Toops et al., 2015; Tan et al., 2016). The results shown in Figure 4 demonstrate that low-dose zoledronic acid decreased cholesterol accumulation in the RPE of Abca4− / − mice.

[0079] Complement-mediated mitochondrial fragmentation Highly differentiated and polarized primary RPE cultures were established from freshly harvested porcine eyes as described (Toops et al., 2014). To induce complement-mediated mitochondrial damage, the RPE monolayer was treated with lipofuscin bisretinoid A2E, which impairs the mechanism protecting RPE from complement activation (Tan et al., 2016). RPE cells were exposed to active complement components (10% normal human serum, NHS, 37 °C for 10 min), which induced complement activation on the RPE cell surface. The inventors have previously shown that this causes mitochondrial fragmentation in RPE (Tan et al., 2016; La Cunza et al., 2021). After exposure to NHS, the cells were treated with 1 μM zoledronic acid for 3 h. RPE mitochondria were labeled with MitoTracker Deep Red, and the cells were live imaged on a Nikon spinning disk confocal microscope as described by the inventors (Tan et al., 2016; La Cunza et al., 2021). The mitochondrial surface was reconstructed from 4D images using Imaris (Bitplane). The results shown in Fig. 5 indicate that zoledronic acid prevented complement-mediated mitochondrial fragmentation in in vitro RPE.

[0080] 12-month-old wild-type mice and Abca4 - / -Mice were administered vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks by intraperitoneal injection (i.p.). At the end of the study, eyes were processed to create RPE flat mounts (after removal of the cornea, lens, vitreous, and retina), and the RPE flat mounts were fixed in paraformaldehyde. The flat mounts were stained with an antibody against TOM20, an outer mitochondrial membrane protein, to label mitochondria. Imaging and reconstruction of mitochondrial volume were performed as shown in Figure 5. The results shown in Figure 6 indicate that low-dose zoledronic acid prevented complement-mediated mitochondrial fragmentation in the RPE of Abca4− / − mice.

[0081] Subretinal microglia 16-month-old wild-type mice and Abca4 - / - Mice were administered vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks by intraperitoneal injection (i.p.). At the end of the study, eyes were enucleated and the cornea, lens, vitreous, and retina were removed. Four to six cuts were made to flatten the eye cup to create RPE flat mounts, which were fixed in 4% paraformaldehyde. The flat mounts were immunostained with an antibody against ionized calcium-binding adapter molecule 1 (Iba1), a protein specifically expressed in microglia. Images were captured on a Nikon spinning disk confocal microscope, and the number of microglia per flat mount was measured. The results shown in Figure 7 demonstrate that low-dose zoledronic acid reduced subretinal microglia in 18-month-old Abca4− / − mice.

[0082] Photoreceptor degeneration 18-month-old wild-type mice and Abca4 - / -Changes in mouse retinal function were determined by recording electroretinograms using a Celeris system (Diagnosys LLC, Lowell, MA) after 8 weeks of vehicle or ZA administration as described above. Scotopic (dark-adapted) ERG responses were measured under flash intensities ranging from 0.158 to 50 cd.s / m 2 . The amplitude of the a-wave, a measure of photoreceptor function, was calculated and plotted as a function of flash intensity. The c-wave, a measure of RPE integrity and function, was recorded while increasing the flash intensity (0.158 to 100 cd.s / m 2 ). The results shown in Figure 8 demonstrate that low-dose zoledronic acid prevented photoreceptor degeneration and visual impairment in 18-month-old Abca4- / - mice.

[0083] Example 3 Treatment of RPE cells in vitro with adiponectin Highly differentiated and polarized primary RPE cultures were established from freshly harvested porcine eyes as described (Toops et al., 2014). To induce complement-mediated mitochondrial damage, the RPE monolayer was treated with lipofuscin bisretinoid A2E, which impairs the mechanism protecting RPE from complement activation (Tan et al., 2016). RPE cells were exposed to active complement components (10% normal human serum, NHS, 37°C for 10 minutes), which induced complement activation on the RPE cell surface. We have previously shown that this causes mitochondrial fragmentation in RPE (Tan et al., 2016; La Cunza et al., 2021). After exposure to NHS, cells were treated with 1 μM adiporon (ADP) for 3 hours. RPE mitochondria were labeled with MitoTracker Deep Red, and cells were live imaged on a Nikon spinning disk confocal microscope as we have described (Tan et al., 2016; La Cunza et al., 2021). The mitochondrial surface was reconstructed from 4D images using Imaris (Bitplane). The results shown in Figure 9 indicate that adiporon prevented complement-mediated mitochondrial fragmentation in RPE in vitro.

[0084] In in vitro studies of hepatocytes, cardiomyocytes, pancreatic cancer cell lines, etc., adiporon has been used at a dose of approximately 25 μM for 24 - 48 hours (in the range of 10 - 50 μM, 25 μM being the effective dose). The experimental results shown in Figure 9 were obtained using 1 μM adiporon for 3 hours. We also used 0.01 μM (data not shown). These data indicate that a 1 / 10 - 1 / 1000 dose (0.01 - 1 μM) for a shorter time (3 hours) is effective against the reduction of ceramide and the prevention of complement-induced mitochondrial damage in primary RPE cultures.

[0085] All patents, patent applications, and publications cited in this specification are hereby incorporated by reference into this specification to the same extent as if each individual patent application or publication was specifically and individually indicated to be incorporated by reference. The disclosed embodiments are provided for purposes of illustration and not limitation. Although the invention has been described with reference to the described embodiments, those skilled in the art will understand that modifications can be made to the structure and elements of the invention generally without departing from the spirit and scope of the invention.

Claims

1. A pharmaceutical composition for treating a retinal disease in a subject requiring treatment of a retinal disease, comprising a therapeutically effective amount of zoledronic acid or a derivative thereof, wherein the zoledronic acid or derivative is administered in an ultra-low dose.

2. The pharmaceutical composition according to claim 1, wherein the retinal disease is a condition mediated by the accumulation of lipofuscin in RPE cells.

3. The pharmaceutical composition according to claim 1, wherein the retinal disease is Stargardt macular dystrophy or dry age-related macular degeneration.

4. The pharmaceutical composition according to claim 1, wherein the retinal disease is selected from the group consisting of neuronal ceroid lipofuscinosis, Batten disease, Vietti crystalline dystrophy, Niemann-Pick disease type C, Doin honeycomb dystrophy, Faber disease, and Best vitiligo macular dystrophy.

5. The pharmaceutical composition according to claim 1, comprising or incorporated in an implant; a drug-eluting device, a drug-eluting structure, or a drug-eluting substance; a polymer-based drug-eluting wafer; an injectable hydrogel; or an implantable hydrogel scaffold.

6. Zoledronic acid or its derivatives (a) administered by intravitreal implant to deliver a dose of 50 ng / day to 50 μg / day to one eye, or (b) administered as an eye drop solution or suspension, or as an ophthalmic ointment or gel, at a dose of 50 ng / day to 50 μg / day per eye, (c) Administered by choroidal injection at a dose of 50 ng / day to 50 μg / day per eye, or (d) administered systemically to provide a dose of 0.001–2.0 mg, 0.01–2.0 mg, or 0.3–0.5 mg. The pharmaceutical composition according to claim 1.

7. Zoledronic acid or its derivatives It is administered systemically in doses selected from the group consisting of 0.001 mg, 0.02 mg, 0.03 mg, 0.04 mg, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.10 mg, 0.20 mg, 0.30 mg, 0.40 mg, 0.50 mg, 0.60 mg, 0.70 mg, 0.80 mg, 0.90 mg, and 1.0 mg, or It is administered systemically in doses of 100 ng to 10 μg per kg of body weight, 1.0 to 7.0 μg per kg of body weight, or approximately 5.0 μg per kg of body weight, or It is administered systemically in amounts of 0.1 μg, 0.2 μg, 0.3 μg, 0.5 μg, 1.0 μg, 2.0 μg, 3.0 μg, 4.0 μg, 5.0 μg, 6.0 μg, 7.0 μg, 8.0 μg, 9.0 μg, or 10.0 μg per kg of body weight. The pharmaceutical composition according to claim 1.

8. The pharmaceutical composition according to claim 1, administered by a route including intravenous delivery, intramuscular delivery, intraperitoneal delivery, or subcutaneous delivery.

9. The pharmaceutical composition according to claim 1, administered at a frequency selected from the group consisting of once a year, once a month, twice a month, once a week, twice a week, every other day, once a day, twice a day, and three times a day.

10. The pharmaceutical composition according to claim 1, comprising zoledronic acid or a derivative thereof, and further comprising any of an excipient, a carrier, a diluent, a release compound, a drug delivery vehicle or a drug targeting vehicle, and an additional active therapeutic agent.

11. The pharmaceutical composition according to claim 10, comprising an adiponectin 1 receptor agonist.

12. The pharmaceutical composition according to claim 11, wherein the adiponectin 1 receptor agonist comprises adipolone.

13. The pharmaceutical composition according to claim 1, wherein zoledronic acid is co-administered with an adiponectin 1 receptor agonist.

14. Zoledronic acid-containing intravitreous implants loaded with 0.001–0.3 mg of zoledronic acid or 0.005–2.5 mg of zoledronic acid.

15. Ophthalmic preparations containing zoledronic acid at a dose of 0.001–0.05 mg / dose or 0.005–0.05 mg / dose, or injectable ophthalmic preparations containing zoledronic acid at a concentration of 1 μg / ml–10 mg / ml or 10 μg / ml–1 mg / ml.

16. The injectable ophthalmic preparation according to claim 15, comprising zoledronic acid conjugated with a dendrimer, or zoledronic acid formulated as nanoparticles.

17. (a) A therapeutically effective amount of adipolone, or (b) A therapeutically effective dose of a bisphosphonate ASM / FDPS inhibitor, A pharmaceutical composition for treating retinal disease in a subject requiring treatment for retinal disease, wherein a bisphosphonate ASM / FDPS inhibitor is administered to the subject at an ultra-low dose.

18. The pharmaceutical composition according to claim 17, wherein the retinal disease is a condition mediated by the accumulation of lipofuscin in RPE cells.

19. The pharmaceutical composition according to claim 17, wherein the retinal disease is Stargard macular dystrophy or dry age-related macular degeneration, or is selected from the group consisting of neuronal ceroid lipofuscinosis, Batten disease, Vietti crystalline dystrophy, Niemann-Pick disease type C, Doin honeycomb dystrophy, Faber disease, and Best vitellate macular dystrophy.

20. Adipolon, or a bisphosphonate-based ASM / FDPS inhibitor, (a) administered in doses of 0.001–2.0 mg, 0.1–1.0 mg, or 0.3–0.5 mg, (b) Administered in a dose selected from the group consisting of 0.001 mg, 0.002 mg, 0.003 mg, 0.004 mg, 0.005 mg, 0.006 mg, 0.007 mg, 0.008 mg, 0.009 mg, 0.01 mg, 0.02 mg, 0.03 mg, 0.04 mg, 0.05 mg, 0.06 mg, 0.07 mg, 0.08 mg, 0.09 mg, 0.10 mg, 0.20 mg, 0.30 mg, 0.40 mg, 0.50 mg, 0.60 mg, 0.70 mg, 0.80 mg, 0.90 mg, and 1.0 mg, or (c) Administered in doses of 100 ng to 10 μg per kg of body weight, 1.0 to 7.0 μg per kg of body weight, or 5.0 μg per kg of body weight, (d) The drug is administered at a dose selected from the following groups per kg of body weight: 0.1 μg, 0.2 μg, 0.3 μg, 0.5 μg, 1.0 μg, 2.0 μg, 3.0 μg, 4.0 μg, 5.0 μg, 6.0 μg, 7.0 μg, 8.0 μg, 9.0 μg, and 10.0 μg. The pharmaceutical composition according to claim 17.

21. The pharmaceutical composition according to claim 17, administered at a frequency selected from the group consisting of once a year, once a month, twice a month, once a week, twice a week, every other day, once a day, twice a day, and three times a day.

22. Whole-body delivery; local delivery; Intravenous delivery; Intramuscular delivery; intraperitoneal delivery; topical delivery; subcutaneous delivery; The pharmaceutical composition according to claim 17, administered by a route comprising either intraocular delivery or topical delivery to the eye.

23. A pharmaceutical composition according to any one of claims 17 to 22, comprising one or more bisphosphonate ASM / FDPS inhibitors or adipolone, and further comprising an excipient, carrier, diluent, release compound, drug delivery vehicle or drug targeting vehicle, and an additional active therapeutic agent.

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