Ultralow dose zoledronic acid for treatment of retinal diseases

Abstract

Provided herein are methods of treating retinal diseases using low doses of bisphosphonate hydrosphingomyelinase (ASM) and a farnesyl diphosphate synthase (FDPS) inhibitor, such as zoledronic acid.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 340,435, filed on May 10, 2022, and U.S. Provisional Application No. 63 / 500,448, filed on May 5, 2023, each of which is incorporated herein by reference for all purposes. Background Art

[0002] The retinal pigment epithelium (RPE) is an important component of the eye. The RPE is composed of a mosaic of flat, hexagonal cells that are tightly bound together at their junctions. On one side of the RPE are adjacent sensory retinal cells that sense light and transmit visual information to the optic nerve. On the other side of the RPE is the choroidal tissue of the eye. The choroidal tissue is a vascularized area that provides water, nutrients, and other compounds to the upper cells of the eye. The RPE plays many key roles in maintaining vision, including isolating the tissues of the eye from the general circulatory system, maintaining a proper ionic environment, processing waste external photoreceptor elements from the neural retinal photoreceptor cells, and protecting the retina from excessive light. Therefore, the RPE is essential for vision by maintaining the photoreceptor cells it supports.

[0003] Various retinal diseases may affect this key part of the eye. The main symptom of RPE is dry age-related macular degeneration (AMD). AMD is a disease that gradually impairs vision in the macula (the central area of ​​the eye). AMD is the leading cause of vision loss in people aged 60 and over. It is estimated that in the United States, 30% of people over the age of 75 suffer from some form of AMD. In dry AMD, accumulation of lipofuscin bisretinoids is observed. These substances are vitamin A metabolites, including undigested photoreceptor outer segment tips (usually digested in the RPE). These lipofuscin bisretinoids gradually accumulate as byproducts of the continuous circulation of retinal chromophores. Another symptom of dry AMD is the accumulation of lipid-protein aggregates (called drusen) above and below the RPE, which destroy the contact with the nutrient choroid below. The gradual accumulation of lipofuscin, bisretinoids and drusen is related to the dysfunction and death of RPE cells.

[0004] Other severe and blinding retinal diseases associated with lipofuscin accumulation are known to afflict millions of patients. For example, Stargardt macular dystrophy, including autosomal dominant Stargardt disease or autosomal recessive Stargardt disease, is a disease characterized by lipofuscin accumulation and macular degeneration. Other retinal diseases include, for example, neuronal ceroidlipofuscinosis, including Batten's Disease and Bestvitelliform macular dystrophy.

[0005] Previous studies have elucidated the pathological cascade of dry AMD and other retinal diseases associated with lipofuscin accumulation, for example, as described by Toops et al. (2015). Cholesterol-mediated activation of acid sphingomyelinase disrupts autophagy in the retinal pigment epithelium (Mol. Biol. Cell 26: 1-14). These studies show that bi-retinoids capture cholesterol and bis (monoacylglycerol) phosphate (an acid sphingomyelinase (ASM) cofactor) within the RPE. The study further showed that this will promote ASM activation, thereby increasing the accumulation of ceramide. Ceramide will promote microtubule acetylation, which will disrupt normal autophagosome transport and impair the important autophagic flux in the RPE. The study also showed that effective autophagy can be restored by inhibiting ASM and promote the health of the RPE by disrupting the pathological cascade triggered by lipofuscin bi-retinoids.

[0006] Therapeutic interventions based on the above findings have been taught, for example, as described in U.S. Patent Application Publication No. US20150366876, entitled "Treatment of Acquired and Inherited Retinal Degeneration Using Acid Sphingomyelinase Inhibitors" filed by Lakkaraju et al. Among them, it was demonstrated that the functional ASM inhibitor, desipramine, can effectively inhibit ceramide accumulation and the pathological cascade it triggers. In addition, the therapeutic use of other ASM inhibitors, including the structural ASM inhibitor zoledronic acid, is suggested, but no relevant proof is provided.

[0007] Despite these recent and promising advances in understanding and treating retinal diseases, clinical applications have yet to be realized and retinal diseases continue to afflict millions of people worldwide. Therefore, there remains a pressing need in the art for effective therapies to treat retinal diseases such as dry AMD, Stargardt's disease, etc.

[0008] As previous studies have shown, ASM inhibitors other than desipramine may be helpful in treating retinal diseases. Zoledronic acid is a known ASM inhibitor and therefore may be helpful in treating retinal diseases. However, to the knowledge of the inventors of the present disclosure, this agent and related ASM bisphosphonate inhibitors have not previously been evaluated for use in retinal diseases. Zoledronic acid also inhibits farnesyl diphosphate synthase (FDPS), which is an enzyme responsible for cholesterol biosynthesis and protein prenylation.

[0009] Zoledronic acid ((1-hydroxy-2-imidazol-1-yl-phosphonoethyl)phosphonic acid monohydrate) is also known as zoledronate, and is a bisphosphonate containing Structure 1: Among the various biological effects of zoledronic acid, as mentioned above, it is an inhibitor of ASM (Roth, et al. Angew Chem Int Ed Engl. 48:7560-7563, 2009), and is also an inhibitor of farnesyl diphosphate synthase (FDPS).

[0010] Zoledronic acid also inhibits bone resorption by inhibiting osteoclast activity and inducing osteoclast apoptosis. Zoledronic acid also binds to bone and prevents osteoclast resorption of mineralized bone. Zoledronic acid is clinically approved for the treatment of osteoporosis and is sold in a variety of forms, such as Aclasta TM (Novartis Pharmaceuticals) and Reclast TM (Novartis Pharmaceuticals). Zoledronic acid is also approved for the treatment of Paget's disease.

[0011] In terms of cancer, zoledronic acid has also been approved for the treatment of bone complications caused by certain cancers, such as malignant hypercalcemia. Zoledronic acid can also induce apoptosis in cancer cells and is approved for the treatment of bone metastases in multiple myeloma and certain solid tumors. In addition, certain types of solid tumors (breast cancer, prostate cancer, lung cancer) can also metastasize to the bone marrow. In the bone marrow, these cancers will destroy the function of osteoclasts, thereby promoting pathological bone resorption and inhibiting the formation of new bone. Zoledronic acid has been approved for the treatment of these conditions because it inhibits osteoclasts and bone resorption and induces apoptosis in cancer cells, and is marketed as Zometa TM (Novartis Pharmaceuticals) name sales.

[0012] In summary, zoledronic acid is considered to be a safe and well-tolerated agent. However, despite the obvious therapeutic uses of zoledronic acid, the agent is associated with various negative side effects. Various sources have pointed out the side effects of zoledronic acid, such as anemia, fatigue, muscle discomfort and swelling of the lower limbs. In addition, some subjects may face the risk of renal impairment due to the use of zoledronic acid, so it may not be recommended for subjects with subnormal renal function (e.g., CKD subjects) to use zoledronic acid. A rare but serious complication of some subjects receiving bisphosphonate treatment is osteonecrosis of the jaw, which mainly occurs in multiple myeloma subjects who receive tooth extraction. In addition, the European Medicines Agency reports that atypical fractures may be a side effect of bisphosphonates. In addition, subjects with hypocalcemia may experience harmful side effects of zoledronic acid.

[0013] As with many therapeutic agents, the administration of bisphosphonates also increases the risk of adverse interactions with other drugs being administered to a subject. For example, adverse interactions between zoledronic acid and drugs such as aspirin and celecoxib can result in an increased risk of gastrointestinal bleeding. Adverse interactions between zoledronic acid and drugs such as acyclovir and cisplatin have been observed to result in an increased risk of nephrotoxicity. Summary of the invention

[0014] The invention disclosed herein is based on the unexpected discovery that bisphosphonate ASM / FDPS inhibitors can be effectively administered to treat retinal diseases at ultra-low doses that have not been considered or suggested in previous use. Specifically, the inventors of the present disclosure have determined that zoledronic acid and related compositions can have therapeutic effects at doses that are much lower (e.g., at least 100 times lower) than the standard dose currently used clinically. Notably, this dose is about 2,000 times lower than the dose of desipramine previously shown to achieve a therapeutic effect in the retina.

[0015] The surprising and unexpected efficacy of these substances at ultra-low doses can provide a variety of benefits. In a first aspect, bisphosphonates (e.g., zoledronic acid) can be used to treat retinal diseases by using them at ultra-low doses while significantly reducing the risk of harmful side effects or adverse drug cross-interactions observed when used at standard doses. Through this risk reduction, the subject population suffering from retinal diseases that can be treated with bisphosphonates is greatly expanded. These and other therapeutic advantages are provided by the methods of the invention disclosed herein.

[0016] In one aspect, the present disclosure relates to a method for treating a retinal disease in an object for treating a retinal disease, the method being by administering to the object a pharmaceutical composition comprising zoledronic acid or a derivative thereof in a therapeutically effective amount, wherein the zoledronic acid or derivative is administered in an ultra-low dose. In some embodiments, the retinal disease is a disease mediated by the accumulation of lipofuscin in RPE cells. In some embodiments, the retinal disease is dry age-related macular degeneration or Stargardt's macular dystrophy. In some cases, the retinal disease is neuronal ceroid lipofuscinosis, Batten's Disease, Bietti's crystalline dystrophy, Niemann-Pick disease type C, Doyne's honeycomb dystrophy, Farber disease or Best vitelliform macular dystrophy. In some embodiments, the pharmaceutical composition comprises or is incorporated into an implant; a drug eluting device, structure or material; a polymer drug eluting wafer; an injectable hydrogel; or an implantable hydrogel scaffold. In some embodiments, zoledronic acid or a derivative is administered via an intravitreal implant to deliver a dose of about 50 ng / day to about 50 μg / day to the eye. In some embodiments, zoledronic acid or a 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, zoledronic acid or a derivative thereof is administered to the eye as an eye drop or suspension, or an ophthalmic ointment or gel at a dose of about 50 ng / day to about 50 μg / day. In some embodiments, zoledronic acid or a derivative is administered to the eye via suprachoroidal injection at a dose of about 50 ng / day to about 50 μg / day. In some embodiments, zoledronic acid or a derivative is systemically administered to provide a dose of 0.001 to 2.0 mg, a dose of 0.01 to 2.0 mg, or a dose of 0.3 to 0.5 mg. In some embodiments, zoledronic acid or a derivative is systemically administered at a dose of 0.3 to 0.5 mg, 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 derivatives are administered systemically, with a dose of 100ng to 10μg per kg of body weight, a dose of 1.0 to 7.0μg per kg of body weight, or a dose of about 5.0μg per kg of body weight. In some embodiments, zoledronic acid or derivatives are administered systemically in an amount of 0.1, 0.2, 0.3, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 or 10.0μg per kg of body weight. In some embodiments, administration is carried out by a route including any one 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, once every other day, once a day, twice a day, and three times a day. In some embodiments, the pharmaceutical composition comprises zoledronic acid and any one of an excipient, a carrier, a diluent, a release formulation, a drug delivery or drug targeting carrier and other active therapeutic agents. In some embodiments, the pharmaceutical composition comprises an adiponectin 1 receptor agonist, such as adiporon. In some embodiments, zoledronic acid is co-administered with an adiponectin 1 receptor agonist.

[0017] In another aspect, the present disclosure provides an intravitreal implant comprising zoledronic acid, wherein the amount of zoledronic acid loaded is 0.001 to 0.3 mg. In some embodiments, the implant is loaded with 0.005 to 2.5 mg of zoledronic acid.

[0018] In another aspect, the present disclosure provides a topical ophthalmic formulation comprising 0.001-0.05 mg / dose of zoledronic acid. In some embodiments, the ophthalmic formulation comprises 0.005-0.05 mg / dose of zoledronic acid.

[0019] In another aspect, the present disclosure provides an injectable ophthalmic formulation comprising zoledronic acid at a concentration of 1 μg / ml-10 mg / ml. In some embodiments, the injectable ophthalmic formulation comprises zoledronic acid at a concentration of 10 μg / ml-1 mg / ml. In some embodiments, the ophthalmic formulation comprises zoledronic acid coupled to a dendrimer or formulated as nanoparticles.

[0020] In another aspect, the present disclosure provides a method for treating a retinal disease in a subject in need of treating a retinal disease, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising adiprolon; or a method for treating a retinal disease in a subject in need of treating a retinal disease, the method comprising 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 disorder mediated by lipofuscin accumulation in RPE cells. In some embodiments, the retinal disease is Stargardt's macular dystrophy or dry age-related macular degeneration. In some embodiments, the retinal disease is neuronal ceroid lipofuscinosis, Batten's Disease, Bietti's crystalline dystrophy, Niemann-Pick disease type C, Doyne's honeycomb dystrophy, Farber disease, or Best vitelliform macular dystrophy. In some embodiments, adiprolon or a bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.001 to 2.0 mg, a dose of 0.1 to 1.0 mg, or a dose of 0.3 to 0.5 mg. In some embodiments, adiprolon or a bisphosphonate ASM / FDPS 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, Adipron or a bisphosphonate ASM / FDPS inhibitor is administered at a dose of 100 ng to 10 μg per kg body weight, 1.0 to 7.0 μg per kg body weight, or about 5.0 μg per kg body weight. In some embodiments, Adipron or a bisphosphonate ASM / FDPS inhibitor is administered at a dose selected from the group consisting of 0.1, 0.2, 0.3, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, and 10.0 μg / kg 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, once every other day, once a day, twice a day, and three times a day. In some embodiments, administration to the eye is by any of the following routes: systemic delivery; topical delivery; intravenous delivery; intramuscular delivery; intraperitoneal delivery; topical delivery; subcutaneous delivery; intraocular delivery; and topical delivery. In some embodiments, the pharmaceutical composition comprises one or more bisphosphonate ASM / FDPS inhibitors or adiprolon and any of an excipient, carrier, diluent, release formulation, drug delivery or drug targeting carrier and other active therapeutic agents. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Multiple pathways targeted by ZA for the prevention of retinal degeneration are delineated. ZA inhibits farnesyl diphosphate synthase (FDPS) and acid sphingomyelinase (ASM), thereby correcting multiple downstream pathological features of macular degeneration.

[0022] Figure 2 Data presented in AD illustrate in vitro screening of ASM inhibitors in RPE. 2A: Acetylated tubulin (green) staining in primary RPE cultures treated with the lipofuscin biretinoid A2E (which activates ASM) ± the ASM inhibitors desipramine (des), amitriptyline (ami), fluoxetine (fluo), or zoledronic acid (ZA). All drugs were used at 10 μM for 3 hours 48 hours after A2E treatment. 2B: Quantification of acetylated tubulin fluorescence in A. *: Significantly higher in drug-treated or control cells, p<0.001. 2C: Live imaging static images of autophagic flux (EGFP-mRFP-LC3) in cells treated with ZA. 2D: EGFP:mRFP ratio as a measure of autophagic flux in primary RPE. ***, p<0.001, compared to cells treated with A2E alone. Fex – fexofenadine; CBZ – cyclobenzaprine.

[0023] Figure 3 Data presented illustrate the effect of ZA treatment on ceramide accumulation in a dry AMD mouse model. Top: 18-month-old Abca4 treated with vehicle or ZA - / - Ceramide staining of RPE flat mounts from mice. Bottom: High magnification images of boxes in flat mounts (all from the subretinal part) and quantification of ceramide in RPE. Actin (highlighted line segments depicted in the field) was stained with phalloidin. Mean ± SEM, *p < 0.05. n ~ 1500-1800 RPE, ≥ 3 mice / group.

[0024] Figure 4 Data presented illustrate the effect of ZA treatment on cholesterol accumulation in a mouse model of macular degeneration. 14 month old wild type and Abca4 - / - Cholesterol in mouse RPE cryosections and quantification. Mean ± SEM, *p < 0.05; n ~ 1500-1800 RPE, ≥ 3 mice per genotype and treatment.

[0025] Figure 5 Data presented illustrate the effect of ZA treatment on complement-mediated mitochondrial fragmentation in RPE in vitro. Representative 3D volume reconstructions of live imaging of Mitotracker-labeled mitochondrial networks in primary polarized RPE monolayers treated or not with A2E and exposed to normal human serum (NHS) as a complement source. Cells were treated with 1 μM ZA for 3 h after complement exposure and imaged immediately. Warmer colors—healthy mitochondria; cooler colors—fragmented mitochondria. Mitochondrial volumes were quantified. Mean ± SEM, *p < 0.05, one-way ANOVA and Tukey's multiple comparison test. n = ~900 cells from 3 independent experiments.

[0026] Figure 6 The data presented illustrate the effect of ZA treatment on complement-mediated mitochondrial fragmentation in a mouse model of macular degeneration. Volume reconstruction and quantification of Tom20-labeled mitochondria in RPE flat mounts of 14-month-old mice (ZA 5 μg / kg, intraperitoneally (ip), three times a week for 8 weeks). Mean ± SEM, *p < 0.05. n = approximately 1500-1800 RPE from ≥ 3 mice / genotype and treatment.

[0027] Figure 7 Data presented show the effect of ZA treatment on subretinal microglia in a mouse model of macular degeneration. Iba1-labeled subretinal microglia (grey) on RPE flat mounts of phalloidin-labeled mice and vehicle-treated 18-month-old Abca4 - / - Quantification of microglia on each mount in ZA mice and ZA mice. Mean ± SEM of subretinal microglia from three mice. *, p < 0.05.

[0028] Figure 8 Data presented show the effect of ZA treatment on photoreceptor degeneration in a mouse model of macular degeneration. Scotopic vision a-wave (reflecting photoreceptor integrity and function) and c-wave (reflecting RPE integrity and function) were measured by electroretinography (Diagnosys Celeris). Mean ± SEM, *p < 0.05. ≥ 3 mice / genotype and treatment.

[0029] Fig. 9 Illustrative data are provided showing the effect of in vitro adipoviron treatment of RPE on complement-mediated mitochondrial fragmentation. Representative 3D volume reconstructions of live imaging of Mitotracker-labeled mitochondrial networks in primary polarized RPE monolayers treated or not with A2E and exposed to normal human serum (NHS) as a source of complement. After exposure to complement, cells were treated with 1 μM adipoviron (ADP) for 3 h and imaged immediately. Mitochondrial volume was quantified. Mean ± SEM, *p < 0.05, one-way ANOVA and Tukey's multiple comparison test. n = approximately 900 cells from 3 independent experiments. DETAILED DESCRIPTION

[0030] As used herein, "a," "an," and "the" include one member but also include members having more than one member unless the context clearly dictates otherwise.

[0031] As used herein, the terms "about" and "approximately" with respect to a given value generally indicate a deviation from the stated value that is generally within 30% or 20% of the stated value. For example, one of ordinary skill in the art recognizes that "about" with respect to a dose or amount provides the same pharmacological effect as that obtained with the specified dose or amount. In certain embodiments, the terms "about" and "approximately" when used in their context include doses or amounts within 20%, within 15%, within 10%, or within 5% of the specified dose or amount.

[0032] The present disclosure provides a method for treating a retinal disease in a subject, the method comprising administering to the subject a pharmaceutical composition comprising a bisphosphonate ASM / FDPS inhibitor in a pharmaceutically effective amount, wherein the bisphosphonate 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 the activity of acid sphingomyelinase (ASM) and farnesyl diphosphate synthase (FDPS). In a related embodiment, the present disclosure also provides a pharmaceutical composition comprising a bisphosphonate ASM / FDPS inhibitor for use in a method for treating a retinal disease in a subject, wherein the method comprises administering a bisphosphonate ASM / FDPS inhibitor at an ultra-low dose. In another aspect, the present disclosure provides a method for manufacturing a drug for treating a retinal disease, comprising using an ultra-low dose of a bisphosphonate ASM / FDPS inhibitor.

[0033] Ultra-low dose. A major aspect of the present invention is the use of bisphosphonate inhibitors of ASM, such as zoledronic acid, at doses far below those currently used clinically. As used herein, "dose" refers to the amount of a selected agent delivered at one time. For example, an example of a sugar dose is "4 grams" or "a 4 gram sugar cube." As used herein, a dose refers to a specific dose of a selected agent delivered within a selected time period, optionally, at a specified number of doses, optionally, at a specified frequency, optionally, by a specified route of administration. For example, an example of a sugar dose is "eight grams of sugar per day, administered in two divided doses of four doses, one in the morning and one at night, orally."

[0034] As used herein, "low dose" for treating retinal diseases with respect to administering ASM / FDPS inhibitors (e.g., zoledronic acid) includes a significantly smaller dose than a clinically commonly used dose, for example, at least 5 times lower, preferably 10 times lower, or 100 times lower bisphosphonate ASM / FDPS inhibitors (e.g., zoledronic acid). In some embodiments, ultra-low doses are at least 10 times lower, preferably 100 times lower, than the clinically commonly used dose of bisphosphonate ASM / FDPS inhibitors (e.g., zoledronic acid). For the treatment of osteoporosis and cancer-related treatments, the standard dose of zoledronic acid is 4 or 5 mg, delivered with a dose including a single dose of 4 or 5 mg zoledronic acid, delivered intravenously once a year. Such administration is on average about 40-100 μg per kilogram of body weight. As described in the Examples section herein, the inventors of the present disclosure have found that, in some embodiments, a dose at least 100 times lower than a given clinical dose can effectively inhibit the pathology mediated by lipofuscin in the RPE caused by various retinal diseases.

[0035] In one embodiment, the methods of the invention include administering a bisphosphonate ASM / FDPS inhibitor (e.g., zoledronic acid) to a human subject at a dose of 0.005 to 10 mg, or 0.01 to 1 mg, for example, at a dose of 0.5 mg to 1.0 mg, for example, at a dose of 0.3 mg to 0.5 mg, for example, in a dose range of 0.01-0.30 mg. For example, exemplary doses include 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. Equivalent doses for smaller human subjects or non-human animal subjects can be calculated based on methods known in the art.

[0036] In one embodiment, the method of the invention comprises administering a bisphosphonate ASM / FDPS inhibitor (e.g., zoledronic acid) at a dose of 10 ng to 50 μg per kg body weight, for example, at a dose of 100 ng to 20 μg per kg body weight, for example, at a dose of 1.0 μg to 10 μg per kg body weight, for example, at a dose of 3-7 μg per kg body weight, or at a dose of about 5 μg per kg body weight. For example, exemplary dosages include 100 ng / kg body weight, 200 ng / kg body weight, 300 ng / kg body weight, 400 ng / kg body weight, 500 ng / kg body weight, 600 ng / kg body weight, 700 ng / kg body weight, 800 ng / kg body weight, 900 ng / kg body weight, 1 μg / kg body weight, 2 μg / kg body weight, 3 μg / kg body weight, 4 μg / kg body weight, 5 μg / kg body weight, 6 μg / kg body weight, 7 μg / kg body weight, 8 μg / kg body weight, 9 μg / kg body weight, and 10 μg / kg body weight.

[0037] Typically, the dose range for bisphosphonate ASM / FDPS inhibitors (eg, zoledronic acid) is 0.001 mg / m2 of body surface area. 2 Up to 1.0 mg / m 2 , for example, the range is 0.037mg / m 2 Up to 0.37 mg / m 2 , for example, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1.0, 0.2, 0.3, 0.4 or 0.5 mg / m 2 The human equivalent dose (HED) of zoledronic acid is 0.1 μg / kg to 1 μg / kg.

[0038] The above disclosed dosage can be given according to any number of suitable dosing regimens, dosages, dosage forms and routes of administration. Regarding frequency, the selected dosage can be given at any selected frequency, such as once a year, once a month, once a week, once a day, etc. The course of treatment can be any course of treatment, for example, one week, two weeks, three weeks, four weeks, two months, three months, six months, one year or indefinitely as needed. Administration can also be carried out on demand as determined by clinical evaluation, or dosage and / or treatment frequency can be adjusted according to clinical evaluation. Additionally or alternatively, the dosage can be increased from a lower dose to at least one higher dose in subsequent administration. In some cases, additionally or alternatively, the dosage can be reduced from a higher dose to at least one lower dose in subsequent administration.

[0039] In addition, administration can be carried out by any selected route of administration. In one embodiment, administration is systemic, for example including intravenous, intraperitoneal, subcutaneous or oral administration. In such embodiments, the bisphosphonate ASM / FDPS inhibitor (for example, zoledronic acid) administered is usually administered with a relatively high dosage compared to topical administration to the eye, so as to reach a therapeutically effective concentration at RPE, for example, the dosage to a human subject is 0.01-1 mg, the dosage is 1-30 μ g per kg of body weight, and the dosage is 0.037-1.0 mg / m 2 .

[0040] In another embodiment, administration includes ocular administration, i.e. administration to the ocular region. For example, selected bisphosphonate ASM / FDPS inhibitor (e.g., zoledronic acid) can be delivered by intraocular administration, for example, by methods such as suprachoroidal injection, subconjunctival injection, intravitreal injection, subretinal or subtenon injection. In such embodiments, the bisphosphonate ASM / FDPS inhibitor administered will bypass the blood-brain barrier (BBB) ​​and systemic dilution and removal, and can reach a therapeutically effective concentration at RPE with a relatively low dosage than systemic administration. For example, in some embodiments, the dosage delivered by intraocular delivery may include 0.001-0.1mg for human subjects, 0.05-2μg per kg of body weight and 0.001-0.1mg per square meter of body surface area.

[0041] In a related implementation, the treatment method using intraocular administration may also include an anti-inflammatory regimen before and after the injection cycle. The anti-inflammatory regimen may include steroidal and non-steroidal anti-inflammatory drugs, alone or in combination. Anti-inflammatory drugs may be administered alone or in combination by oral, intraocular and / or topical routes.

[0042] In a related implementation, the administration to the eye is to be administered topically to the eye, for example, by using a gel, an eye ointment or drops to be applied to the outer surface of the eye. Exemplary dosage forms include eye drops or gels, for example, solutions, suspensions, emulsions or other preparations of the selected bisphosphonate ASM / FDPS inhibitor (for example, zoledronic acid) included in the vehicle. In such implementations, it is contemplated that delivery will be more effective than systemic delivery, but less efficient than intraocular delivery, because reagent must pass through vitreous membranes and eye fluid and photoreceptors to reach RPE. In some embodiments, the topical dose applied to the eye may include 0.01-0.5 mg for human subjects, 0.5-8 μ g per kg of body weight and 0.0185-0.3 mg per square meter of body surface area.

[0043] In some embodiments, the human dose is based on the animal dose, for example, based on dose scaling conversions commonly used in the art (see, for example, Nair & Jacob, J. Basic and Clin. Pharmac 2016; 7: 27-31). Thus, for example, the animal dose in mg / kg can be converted to a human equivalent dose in mg / kg by dividing the animal dose by 12.3.

[0044] In some embodiments, zoledronic acid is formulated into an ophthalmic solution or suspension for administration in the form of eye drops, for example, as a solution, suspension, liposomal formulation, mucoadhesive polymer, micelle, nanoparticle, 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 eye 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 100 ng / day, 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 not more than about 50 μg / day. In some embodiments, eye drops are formulated to provide a constant intraocular concentration of at least 100pM, 1nM, 10nM, 100nM, 200nM, 500nM or 1μM zoledronic acid. In some embodiments, an ophthalmic solution is administered once a day to provide a daily indicated dose. In some embodiments, more than one (e.g., twice or three times a day) ophthalmic solution is administered daily to provide a daily indicated dose. In some embodiments, an ophthalmic solution can be administered at least once a day, twice a week, three times a week, four times a week or more. In some embodiments, zoledronic acid can be administered once a week or once every two weeks. In some embodiments, zoledronic acid can be administered once a month or twice a month.

[0045] In some embodiments, zoledronic acid is delivered to the eye using an intravitreal implant. In such embodiments, zoledronic acid can be incorporated into an implant to provide zoledronic acid release over a period of up to several years, for example, up to 2 or 3 years. In some embodiments, the implant can be loaded with about 0.001 to about 0.3 mg of zoledronic acid. In some embodiments, zoledronic acid can be incorporated into an implant to release zoledronic acid in a period of less than a few weeks to a few months, for example, 2-4 weeks, 4-6 weeks, 6-8 weeks, 3-6 months, 6-12 months, or 12-18 months. In some embodiments, the implant can be a biodegradable polymer. In some embodiments, the implant is loaded with a certain amount of zoledronic acid to provide a release of about 0.001 μg / day to about 2.5 μg / day dosage, or about 0.005 μg / day to about 2.0 μg / day dosage. 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 of about 10 ng / day to about 250 μg / day. In some embodiments, the implant is loaded with zoledronic acid to provide a dose of about 50 ng / day to about 100 μg / day of zoledronic acid. In some embodiments, the implant is loaded with zoledronic acid to provide a dose of about 100 ng / day to about 50 μg / day of zoledronic acid. In some embodiments, the implant is loaded with zoledronic acid to provide a dose of about 100 ng / day to about 10 μg / day of zoledronic acid. In some embodiments, the implant is loaded with zoledronic acid to provide a dose of zoledronic acid 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, but not more than about 50 μg / day or 100 μg / day. In some embodiments, the sustained release implant is an immiscible or partially immiscible liquid, including but not limited to benzyl benzoate and / or silicone oil. Such embodiments may comprise a suspension, emulsion or solution of zoledronic acid.

[0046] In some embodiments, zoledronic acid is administered using suprachoroidal injection. In some embodiments, a zoledronic acid pharmaceutical composition is used, wherein zoledronic acid is coupled to a dendrimer (see, e.g., Pitha et al., Biomacromolecules 24: 1355-1365, 2023) or provided in a nanoparticle formulation (see, e.g., Pitha et al., Biomacromolecules 24: 1355-1365, 2023; Laradji et al., Polymers 13 (19), 3324, 2021). In some embodiments, the injectable solution or suspension is formulated to provide a concentration of 1 μg / ml-10 mg / ml of zoledronic acid and is delivered by suprachoroidal injection or intravitreal injection. In some embodiments, an injectable solution or suspension comprising zoledronic acid, e.g., coupled to a dendrimer or in a nanoparticle formulation, is formulated to deliver zoledronic acid in an amount of about 0.1-1 mg / eye. In some embodiments, the amount of zoledronic acid delivered to the eye by injection (e.g., suprachoroidal or intravitreal injection) is a dose of about 10 ng / day to about 50 μg / day. In some embodiments, the amount of zoledronic acid delivered to the eye by injection (e.g., suprachoroidal or intravitreal injection) is a dose of about 10 ng / day to about 50 μg / day. In some embodiments, the dose of zoledronic acid for injection is about 100 ng / day to about 50 ug / day. In some embodiments, the dose of zoledronic acid for injection is about 100 ng / day to about 10 μg / day. In some embodiments, the injection is loaded to provide a zoledronic acid dose 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, but not more than about 100 μg / day, preferably 50 μg or less / day. In some embodiments, the injectable composition (eg, for suprachoroidal or intravitreal injection) is formulated 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.

[0047] In some embodiments, the ophthalmic or systemic compositions may be administered using a pre-filled syringe.

[0048] Bisphosphonate ASM / FDPS inhibitors. The methods of the present invention include using bisphosphonate ASM / FDPS inhibitors to treat selected retinal diseases. In the main embodiment, the bisphosphonate ASM / FDPS inhibitor is zoledronic acid or a pharmaceutically effective variant thereof. As used herein, zoledronic acid includes any therapeutically active form of zoledronic acid, including structure 1, its pharmaceutically effective salts, its anhydrous, hygroscopic and hydrous forms, its lipophilic derivatives and chemically related derivatives thereof.

[0049] In one embodiment, zoledronic acid is provided in the form of a salt, such as any one of arginine salts, calcium salts, chromium salts, citrullinate salts, cobalt salts, copper salts, creatine salts, glutamine salts, histidine salts, iron salts, isoleucine salts, leucine salts, lithium salts, lysine salts, magnesium salts, manganese salts, molybdenum salts, ornithine salts, potassium salts, selenium salts, sodium salts, zinc salts, and combinations thereof.

[0050] In one embodiment, the bisphosphonate ASM / FDPS inhibitor includes a derivative of zoledronic acid, such as disclosed by Jaeggi and Wilder in U.S. Patent No. 4,939,130, "Substituted alkanediphosphonic acids and pharmaceutical use". In one embodiment, the bisphosphonate ASM / FDPS inhibitor includes a zoledronic acid derivative, such as disclosed by Hanna et al. in PCT International Patent Application Publication No. WO / 2012071517, "Novel Crystalline Forms". In one embodiment, the bisphosphonate ASM inhibitor includes a zoledronic acid derivative, such as disclosed by Glausch et al. in U.S. Patent Application Publication No. US20100056481, "Crystalline forms of zoledronic acid". In another embodiment, the bisphosphonate ASM / FDPS inhibitor includes a zoledronic acid derivative coupled to deoxycholic acid linked to lysine to increase oral absorption, such as described in Jeon et al., 2016. In another embodiment, the bisphosphonate ASM / FDPS inhibitor includes the compositions described in US Pat. No. 9,682,091 and entitled "Oral Forms of a Phosphonic Acid Derivative."

[0051] In one embodiment, the bisphosphonate ASM / FDPS inhibitor includes a form of zoledronic acid configured for oral delivery. Oral delivery conveniently avoids the inconvenience and risk of morbidity of ocular injections. For example, in one embodiment, the bisphosphonate ASM / FDPS inhibitor includes an orally bioavailable form of zoledronic acid as disclosed in the following documents: U.S. Patent Application Publication No. US20140051669 of Tabuteau et al. "Compositions of Zoledronic Acid or Related Compounds for Treating Disease"; or PCT International Patent Application Publication No. WO2013015599 of Kim et al. "Pharmaceutical Composition for Oral Administration Comprising Bisphosphonic Acid or its Salt".

[0052] Adipron. In another aspect of the present disclosure, Adipron (PubChem CID 16307093) is used to treat selected retinal diseases. Adipron (Case number) is a selective, orally active, synthetic small molecule agonist of adiponectin receptor 1 (AdipoR1) and adiponectin receptor 2 (AdipoR2) synthetic adiponectin receptor agonists. In some embodiments, Adipron is administered in an amount described herein for bisphosphonate ASM / FDPS inhibitors. In some embodiments, the dosage of Adipron is 10-1000 times lower than the dosage for other indications.

[0053] Treatment of Retinal Diseases. The methods of the present invention are directed to treating retinal diseases in a subject in need of treatment for retinal diseases by administering a low dose of a bisphosphonate ASM / FDPS inhibitor.

[0054] With respect to the subject, the subject can be any animal subject in need of treatment for a retinal disease. The subject can be any animal species. In a primary embodiment, the subject is a human, such as a human patient. In other implementations, the subject is a non-human animal, such as a veterinary subject, a pet, livestock, or a test animal. Exemplary non-human animals include mice, rats, pigs, horses, cows, dogs, cats, non-human primates, and the like.

[0055] The subject can be a subject suffering from or diagnosed with a selected retinal disease. The subject can be a subject suspected of suffering from a selected retinal disease. In one embodiment, the subject is a subject at risk of developing a selected retinal disease. In one embodiment, the subject is an elderly subject, for example, a human subject at least 50 years old, at least 55 years old, at least 60 years old, or at least 65 years old. In one embodiment, the subject is a subject with one or more genetic markers indicating a risk of a retinal disease, or a subject with a family history of a retinal disease and a genetic risk. In one implementation, the subject is at risk for, suspected of suffering from, or suffering from dry age-related macular degeneration.

[0056] In some cases, the subject is a human child, adolescent, or young adult, e.g., 25 years of age or younger, who is at risk for, suspected of having, or diagnosed with a retinal disease. In embodiments, the subject is at risk for, suspected of having, or having Stargardt's macular dystrophy, also referred to herein as Stargardt's disease. In some embodiments, the subject is an adult over 25 years of age who is at risk for, suspected of having, or having a retinal disease. In some embodiments, the subject has autosomal dominant Stargardt's disease. In some cases, the subject has autosomal recessive Stargardt's disease.

[0057] For retinal disorders that can be treated by the inventive method, retinal disorders can be any retinal disorders, wherein known or suspected to have RPE dysfunction, microglia activation or photoreceptor defect, including any disorder characterized by or related to the following content: lipofuscin accumulation in RPE cells, cholesterol accumulation in RPE cells, ASM or FDPS abnormal activation in RPE cells, abnormal production of ceramide in RPE, Rab GTPase prenylation abnormality in RPE, abnormal acetylation of microtubules in RPE cells, impaired autophagy in RPE cells, accumulation of drusen or similar deposits on or below RPE, complement-mediated mitochondrial damage in RPE, the presence of subretinal microglia, or photoreceptor loss or functional defects of photoreceptors. As previously mentioned, in the main embodiment, the disorder is dry AMD. In other embodiments, the retinal disease is Stargardt's macular dystrophy, for example, including autosomal dominant or autosomal recessive Stargardt's disease; Dohlmann's honeycomb dystrophy; acid ceramidase diseases, such as Farber's disease; cholesterol and ceramide accumulation and autophagy defect diseases, such as Niemann-Pick disease type C; and diseases such as (neuronal ceroid lipofuscinosis), Best's vitelliform macular dystrophy; retinitis pigmentosa; and Bitt's crystalline dystrophy.

[0058] The method of the present invention includes the treatment of selected retinal diseases. As used herein, "treatment" will cover the treatment effects and results of any number of treatments related to selected retinal diseases, including, for example: improving the symptoms related to selected retinal diseases; slowing down the progress of selected retinal diseases; preventing selected retinal diseases from causing further damage to RPE; improving RPE function; maintaining, improving or restoring photoreceptor health; maintaining, improving or restoring vision; or any other reduction in the incidence rate associated with selected retinal diseases. Treatment used herein will further include preventing selected retinal diseases. For example, preventive treatment can include achieving any of the following goals: preventing or delaying the onset of selected retinal diseases of high-risk subjects; maintaining normal vision or function of RPE; or otherwise preventing the onset of selected retinal diseases. As used herein, treatment also includes preventing progression to geographic atrophy, slowing progression of geographic atrophy, treating neovascularization, preventing progression of neovascular AMD, reducing the number and / or frequency of other AMD treatments (e.g., anti-VEGF injections, complement inhibitors, etc.), changing the total area of ​​geographic atrophy (GA) based on fundus autofluorescence, reducing photoreceptor loss (e.g., as measured by elliptical zone attenuation area), changing reading speed in standard and low light, changing contrast sensitivity, changing retinal sensitivity on visual field examination, changing patient-reported outcomes, changing dark adaptation, changing drusen volume, changing pigmented mottling, significant macular atrophy, bull's eye maculopathy, or fundus spots, changing electroretinogram (ERG) and / or structural changes measured by optical coherence tomography (OCT).

[0059] Treatment also includes any inhibition of the potential pathological process of selected retinal diseases.For example, exemplary therapeutic effect includes reducing the lipofuscin accumulation in RPE, reducing the cholesterol accumulation in RPE cells, reducing the ASM and / or FDPS activity in RPE cells, reducing abnormal ceramide production, reducing abnormal Rab prenylation, reducing the abnormal acetylation of microtubules in RPE cells, improving or recovering the autophagy ability, activity and flux in RPE cells, reducing the formation of drusen accumulation or similar deposits, reducing the mitochondrial damage of complement mediation in RPE, preventing or saving microglia from migrating to the subretinal space, or preventing or saving photoreceptor loss and functional defects.As shown in Example 3 herein, zoledronic acid advantageously reduces the accumulation of lipofuscin, cholesterol and ceramide to the inhibition of bisphosphonates ASM and FDPS, which is contrasted with the medicine such as desipramine that only reduces ceramide studied before.Therefore, in one embodiment, the scope of the present invention also includes reducing the method for lipofuscin accumulation, cholesterol accumulation and / or ceramide accumulation by giving zoledronic acid or its derivatives (for example, with ultra-low dose).

[0060] The treatment of the present invention includes administering to the subject a therapeutically effective amount of one or more bisphosphonate ASM / FDPS inhibitors. In one measurement, a therapeutically effective amount is an amount of a bisphosphonate ASM / FDPS inhibitor sufficient to induce a measurable therapeutic effect. The therapeutic effect can be to achieve a selected physiological result or state, for example, to reduce lipofuscin, cholesterol or ceramide accumulation; reduce ASM and FDP activity; improve autophagy flux in RPE; reduce drusen formation of RPE cells; reduce complement-mediated mitochondrial damage in RPE cells; improve the health or function of RPE; improve the health or function of photoreceptors; reduce the activation of microglia; and improve vision. As used herein, "subject" refers to any mammal, including non-human primates, mice, rats, rabbits, pigs, horses, cattle, goats, sheep, dogs, cats, etc. In a preferred embodiment, the subject is a human.

[0061] Pharmaceutical compositions. The methods and compositions of the present invention include administering one or more bisphosphonate ASM / FDPS inhibitors, such as zoledronic acid. One or more bisphosphonate ASM / FDPS inhibitors can be formulated as a "pharmaceutical composition". As used herein, a pharmaceutical composition will contain one or more bisphosphonate ASM inhibitors and may also contain any number of other material compositions, including excipients, carriers, diluents, release formulations, drug delivery or drug targeting carriers, and other active therapeutic agents. The pharmaceutical compositions of the present invention can be formulated to be compatible with a selected route of administration, such as oral, injection or ocular delivery.

[0062] The pharmaceutical composition of the present invention may include one or more bisphosphonate ASM / FDPS inhibitors, such as zoledronic acid, combined with a drug delivery composition. The drug delivery composition includes any part, material or other material composition, which helps to deliver the ASM / FDPS inhibitor (for example, zoledronic acid) to RPE. In some implementations, the drug delivery composition helps to target the ASM / FDPS inhibitor (for example, zoledronic acid) to other selected target compartments of CNS, PNS or nervous system. In some implementations, the delivery composition includes the material composition promoting across the blood-brain barrier. In some embodiments, the delivery composition includes the material composition promoting across the blood-retinal barrier (BRB).

[0063] The pharmaceutical composition can comprise any form of combination, including functionalization of the bisphosphonate ASM / FDPS inhibitor (e.g., zoledronic acid) with the delivery composition, coupling of the bisphosphonate ASM / FDPS inhibitor with the delivery composition; mixing of the bisphosphonate ASM / FDPS inhibitor with the delivery composition; encapsulation or infusion of the bisphosphonate ASM / FDPS inhibitor within the delivery composition, or any other combination.

[0064] In one implementation, the delivery composition includes a material that promotes bisphosphonate ASM / FDPS inhibitors (e.g., zoledronic acid) to cross the BBB or BRB. For example, the delivery composition may include a ligand that promotes transcytosis through brain endothelial cells across the BBB to the basolateral side, such as an anti-transferrin receptor antibody or its antigen-binding fragment, such as an OX26 antibody, Angiopep2 or similar polypeptide, ApoE protein and its mimetics; diphtheria toxin and a surfactant. Other targeting moieties include BBB-crossing peptides, such as those described in 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 hypertonic or hyperosmotic preparations to promote BBB or BRB transport.

[0065] In one implementation, the delivery composition includes a carrier, which is coupled to a bisphosphonate ASM / FDPS inhibitor (e.g., zoledronic acid), encapsulated therein or otherwise combined with it, to promote crossing the BBB and / or delivery to target RPE cells. Exemplary carriers include: liposomes; extracellular vesicles or their synthetic mimics, such as exosomes; red blood cells modified with bisphosphonate ASM / FDPS inhibitors; microspheres, such as poly (lactic acid-co-glycolic acid) (PLGA) microspheres; and other drug delivery nanoparticles, such as PLGA-PEG nanoparticles, alginate or chitosan nanoparticles, silica nanoparticles and iron oxide nanoparticles. Carrier molecules or compositions can be further functionalized with ligands, and the ligands promote the crossing of BBB or BRB, such as anti-tfR antibodies, Angiopep2 or similar polypeptides, ApoE proteins and their mimics; diphtheria toxin and surfactants, as described above.

[0066] In some embodiments, the bisphosphonate ASM / FDPS inhibitor (eg, zoledronic acid) is administered as a drug-antibody conjugate, eg, comprising an antibody or antigen-binding fragment thereof that targets a ligand present on the RPE.

[0067] In one embodiment, the delivery composition includes, for example, a drug eluting implant placed in a target tissue (e.g., eyes) or is included in an implant. For example, exemplary implants include implants made of biodegradable materials, such as PLGA, polymer drug eluting wafers, polymer drug eluting rods, injectable hydrogels, implantable hydrogel scaffolds, systems based on hydrophilic microspheres, systems based on cyclodextrins, systems based on polymer micelles, and other drug eluting implants known in the art. The exemplary implants for reagent delivery to RPE include implants described in the following documents: Tao et al. WO2012177968 "A scaffold for subretinal cell transplantation and drug delivery" ". Exemplary polymers and implants for ocular drug delivery are also by Allyn et al., Frontiers in Medicine, 8: Article 787644, January 2022, Cao et al., Drug Discovery Today 24 (8), 1694-1700 2019).

[0068] In one implementation, the delivery composition includes an intravitreal implant comprising zoledronic acid, wherein the zoledronic acid is loaded in an amount of 0.001-0.3 mg. In some embodiments, the implant is loaded with 0.005 to 2.5 mg of zoledronic acid.

[0069] In other embodiments, the delivery composition comprises a topical ophthalmic formulation comprising 0.001-0.05 mg / dose of zoledronic acid. In some embodiments, the formulation comprises 0.005-0.05 mg / dose of zoledronic acid. In some embodiments, the topical ophthalmic formulation is a solution or suspension administered as eye drops.

[0070] In other embodiments, the delivery composition comprises an injectable ophthalmic formulation, e.g., suitable for suprachoroidal or intravitreal injection, comprising zoledronic acid at a concentration of 1 μg / ml to 10 mg / ml. In some embodiments, the injectable ophthalmic formulation comprises zoledronic acid at a concentration of 10 μg / ml-1 mg / ml. In some embodiments, the injectable ophthalmic formulation comprises zoledronic acid coupled to a dendrimer or formulated as nanoparticles.

[0071] It should be understood that the methods of the present invention also include the co-administration of a bisphosphonate ASM / FDPS inhibitor (e.g., zoledronic acid) and one or more other active agents for treating retinal diseases. For example, other treatments for treating macular degeneration or Starsgardt's disease include, but are not limited to, anti-VEGF therapy (including, but not limited to, rolucizumab, aflibercept, ranibizumab, pegaptanib sodium, faricimab-svoa, and bevacizumab), complement inhibitors (including, but not limited to, pegcetacoplan and avacincaptad pegol), MCO 010, deuterated retinol, emixustat, MA09 hRPE, STG-001, tinlarebant, QR1011, and REV0100. As used herein, co-administration may include any combination of bisphosphonate ASM / FDPS inhibitor administration and other treatments for one or more retinal diseases. For example, one skilled in the art can determine the timing of administration of ASM / FDPS inhibition therapy and one or more other treatments. In various implementations, ASM / FDPS inhibition therapy and one or more other treatments are administered simultaneously, sequentially, or alternately. In one embodiment, the first and second treatments are administered simultaneously, i.e., simultaneously or overlapping in time. In one embodiment, the first and second treatments are administered in a pharmaceutical composition comprising a combination product comprising a bisphosphonate ASM / FDPS inhibitor and other agents, for example, administered in a single dose form. In one embodiment, one or more other treatments include administering an adiponectin 1 receptor agonist, for example, as described in Kim et al., 2022. Adiponectin receptor agonists improve cardiac lipotoxicity by enhancing ceramide metabolism in type 2 diabetic mice. Cell Death&Disease 13:282. In one embodiment, the adiponectin 1 receptor agonist is adiprolon. In one embodiment, the pharmaceutical compositions of the invention include combination products comprising a bisphosphonate ASM / FDPS inhibitor in combination with one or more adiponectin 1 receptor agonists (eg, adiprolon).

[0072] The pharmaceutical compositions of the present invention can be formulated into a variety of dosage forms. Exemplary dosage forms include: liquid solutions; sachets, capsules or tablets, each containing a predetermined amount of active ingredients, as solids or particles; liquid suspensions; emulsions; aqueous and non-aqueous solutions; isotonic sterile injection solutions; compositions stored under freeze-dried, lyophilized conditions; and other dosage forms known in the art. In one embodiment, the formulation is for topical application to the eye, including eye drops, eye ointments or gels.

[0073] Exemplary implementations include, but are not limited to, the following: Embodiment 1: A method of treating a retinal disease in a subject in need of treatment for the 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. Embodiment 2: The method of embodiment 1, wherein the retinal disease is a condition mediated by lipofuscin accumulation in RPE cells. Embodiment 3: The method as described in Embodiment 1, wherein the retinal disease is dry age-related macular degeneration. Embodiment 4: A method as described in embodiment 1, wherein the retinal disease is selected from the group consisting of Stargardt's macular dystrophy, neuronal ceroid lipofuscinosis, Batten's disease, Bitt's crystalline dystrophy, Niemann-Pick disease type C, Dohlmann's honeycomb dystrophy, Farber's disease, and Best's vitelliform macular dystrophy. Embodiment 5: The method according to embodiment 1, wherein the bisphosphonate ASM / FDPS inhibitor is zoledronic acid or a derivative thereof. Embodiment 6: The method of embodiment 1, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.005 to 2.0 mg. Embodiment 7: The method of embodiment 6, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.1 to 1.0 mg. Embodiment 8: The method of embodiment 7, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.3 to 0.5 mg. Embodiment 9: A method as described in embodiment 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. Embodiment 10: The method according to embodiment 1, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 100 ng to 10 μg per kg body weight. Embodiment 11: The method of embodiment 10, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 1.0 μg to 7.0 μg per kg body weight. Embodiment 12: The method of embodiment 11, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose of about 5.0 μg / kg body weight. Embodiment 13: A method as described in embodiment 10, wherein the bisphosphonate ASM / FDPS inhibitor is administered at a dose selected from the group consisting of 0.1, 0.2, 0.3, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0 μg / kg body weight. Embodiment 14: The method of any one of embodiments 1-13, 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, once every other day, once a day, twice a day, and three times a day. Embodiment 15: A method as described in any of embodiments 1-4, wherein the administration is given by a route including any of the following: systemic delivery; local delivery; intravenous delivery; intramuscular delivery; intraperitoneal delivery; local delivery; subcutaneous delivery; intraocular delivery; suprachoroidal delivery; and local delivery. Embodiment 16: A method as described in any of Embodiments 1-15, wherein the pharmaceutical composition comprises one or more bisphosphonate ASM / FDPS inhibitors and any of an excipient, carrier, diluent, release formulation, drug delivery or drug targeting carrier and other active therapeutic agents. Embodiment 17: The method of embodiment 16, wherein the pharmaceutical composition comprises an adiponectin 1 receptor agonist. Embodiment 18: The method of embodiment 17, wherein the adiponectin 1 receptor agonist comprises adiprolon. Embodiment 19: A method as described in any one of embodiments 1-18, wherein the pharmaceutical composition comprises any one of the following: a solution; a liquid suspension; an emulsion; an isotonic sterile injection solution; a composition in a lyophilized state; an eye drop; or an eye ointment or gel. Embodiment 20: A method as described in any of embodiments 1-19, wherein the drug composition includes or is incorporated into an implant; a drug eluting device, structure or material; a polymer drug eluting chip; an injectable hydrogel; and an implantable hydrogel scaffold. Embodiment 21: The method of embodiment 1, wherein the bisphosphonate ASM is co-administered with an adiponectin 1 receptor agonist. Embodiment 22: The method of embodiment 21, wherein the adiponectin 1 receptor agonist is adipolone. Example Example 1. In vitro screening of ASM inhibitors

[0074] A study was initiated to evaluate ASM inhibitors for the treatment of retinal diseases associated with pathological lipofuscin accumulation.

[0075] Primary RPE cell cultures were established with cells from pig eyes. Cultured RPE cells were treated with lipofuscin bi-retinoid A2E, which has been previously shown to activate ASM. The cultures were then treated with various ASM inhibitors and the measurements of lipofuscin-mediated damage were assessed. The ASM inhibitors were desipramine, amitrpytline, fluoxetine, and zoledronic acid, and the various inhibitors were applied at 10 μM and added to the cultures 48 hours after A2E treatment.

[0076] Acetylated tubulin in cultured RPE cells was assessed by immunostaining with specific antibodies. Figure 1As previously described in Toops et al. (2015), lipofuscin accumulation traps cholesterol within RPE cells, triggering abnormal activation of ASM. ASM activity leads to the accumulation of ceramide, which in turn promotes the acetylation of microtubules. This microtubule acetylation is unstable and inhibits RPE autophagy. The basic function of RPE is to autophagically degrade cellular debris and remove photoreceptor outer segment tips from overlying photoreceptor cells. Microtubule acetylation disrupts normal autophagosome movement and the interaction between autophagosomes and lysosomes, which are essential for the autophagic activity of RPE. This leads to the accumulation of intracellular debris in the RPE, which can induce metabolic stress and cause the formation of drusen in the RPE. Microtubule acetylation also makes the RPE susceptible to complement-mediated mitochondrial damage, leading to the formation of destructive aggregates of drusen. Therefore, the abundance of acetylated microtubules in the RPE provides a measure of lipofuscin-induced damage and the activation of the above pathological cascades.

[0077] Compared with RPE cells not exposed to A2E, primary RPE cultures treated with A2E showed a significant increase in acetylated tubulin, indicating impaired autophagy and resulting damage ( Figure 2 B) Furthermore, the use of an ASM inhibitor after A2E treatment significantly reversed this A2E-induced effect. Among the ASM inhibitors tested, zoledronic acid was the most effective in reducing the abundance of acetylated tubulin.

[0078] RPE cells were treated with A2E and then with ASM inhibitors, and autophagic flux was further assessed by live cell imaging. An autophagy reporter gene (tfLC3) containing the autophagy target microtubule-associated protein 1A / 1B-light chain 3 (LC3) fused to tandem red fluorescent protein (mRFP) and enhanced green fluorescent protein (EGFP) was administered to cultured RPE cells. The uptake of tfLC3 by autophagosomes and its delivery to lysosomes results in acidic quenching of the EGFP signal of the reporter gene, so simultaneous monitoring of mRFP and EGFP signals provides a method for observing and quantifying autophagosomes in RPE cells, where a reduced ratio of EGFP signal to mRFP signal indicates autophagic activity.

[0079] like Figure 2 As shown in D, the ratio of EGFP:mRFP in RPE cells treated with A2E was significantly increased relative to control cells not exposed to A2E, indicating impaired autophagy. Administration of an ASM inhibitor restored autophagy in A2E-treated cells. The most potent autophagy promoter was zoledronic acid. Example 2. Development of Ultra-Low Zoledronic Acid Doses

[0080] Based on the results described in Example 1, we next evaluated the in vivo effects of zoledronic acid on lipofuscin-induced RPE damage. - / - Mice. Autosomal recessive Stargardt's disease is a disorder that involves macular degeneration characterized by accumulation of lipofuscin in the RPE. This form of Stargardt's disease is caused by mutations in the ABCA4 gene. - / - Mice provide a model for studying dry AMD, Stargardt's disease, and other conditions in which pathological lipofuscin accumulation occurs.

[0081] In preliminary in vivo studies, mice were given a single intraperitoneal injection of zoledronic acid. The dose selected was 1 mg per kilogram of body weight, or about 28 μg per mouse. This dose was selected based on the approved dose of zoledronic acid for the treatment of osteoporosis and cancer in humans. Although the use of zoledronic acid is known to produce adverse side effects, the drug is generally considered to be well tolerated. However, when mice were given a dose of 1 mg per kilogram of body weight, significant adverse effects were observed. Side effects included osteonecrosis of the jaw, difficulty eating, and other symptoms such as weakness and lethargy.

[0082] Based on the above results, the effect of lower doses of zoledronic acid on Abca4 - / - The role of mouse. - / - Mice were given a single intraperitoneal injection of zoledronic acid at a dose of 5 μg / kg body weight, a dose 100-fold lower than that used to treat osteoporosis and cancer. Ceramide accumulation

[0083] Wild-type and Abca4 cells were injected intraperitoneally (ip) to 16-month-old mice. - / - Mice were given vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks. At the end of the study, the eyes were enucleated and the cornea, lens, vitreous, and retina were removed. Four to six relaxing incisions were made in the eyecup to generate RPE flat mounts and fixed with 4% paraformaldehyde. Ceramide levels were measured by immunostaining with a specific anti-ceramide antibody (Enzo), which has been extensively validated by us 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 ceramide fluorescence intensity of each cell was quantified using Imaris (Bitplane). Figure 3 The results shown demonstrate that administration of low doses of zoledronic acid reduces the accumulation of ceramide. Cholesterol accumulation

[0084] Wild-type and Abca4 cells were injected intraperitoneally (ip) to 12-month-old mice. - / -Mice were given vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks. At the end of the study, the cornea, lens, and vitreous were removed and embedded in eyecups for cryosectioning. 10 μm thick retinal cryosections were stained with filipin (Sigma), which specifically binds to free cholesterol (Toops et al., 2015; Tan et al., 2016). Figure 4 The results shown in the literature indicate that low-dose zoledronic acid reduces Abca4 - / - Cholesterol accumulation in mouse RPE. Complement-mediated mitochondrial fragmentation

[0085] Highly differentiated polarized primary RPE cultures were established from freshly harvested porcine eyes as described by Toops et al., 2014. To induce complement-mediated mitochondrial damage, RPE monolayers were treated with the lipofuscin diretinoid A2E, which impairs the mechanisms that protect RPE from complement activation (Tan et al., 2016). RPE cells were exposed to active complement components (10% normal human serum, NHS, for 10 min at 37°C) to induce complement activation on the surface of RPE cells. We have previously shown that this leads to mitochondrial fragmentation in RPE (Tan et al., 2016; La Cunza et al., 2021). Following NHS exposure, cells were treated with 1 μM zoledronic acid for 3 h. RPE mitochondria were labeled with MitoTracker Deep Red, and cells were imaged live on a Nikon spinning disk confocal microscope as we have described (Tan et al., 2016; La Cunza et al., 2021). Mitochondrial surfaces were reconstructed from 4D images using Imaris (Bitplane). Figure 5 The results shown in indicate that zoledronic acid prevents complement-mediated mitochondrial fragmentation in RPE in vitro.

[0086] Wild-type and Abca4 cells were injected intraperitoneally (ip) to 12-month-old mice. - / - Mice were given vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks. At the end of the study, eyes were processed to generate RPE flat mounts (after removing the cornea, lens, vitreous, and retina) and fixed in paraformaldehyde. Flat mounts were stained with antibodies against the mitochondrial outer membrane protein TOM20 to mark mitochondria. Figure 5 Mitochondrial volumes were imaged and reconstructed as shown. Figure 6 The results shown in indicate that low-dose zoledronic acid prevents complement-mediated mitochondrial fragmentation in the RPE of Abca4- / - mice. Subretinal microglia

[0087] Wild-type and Abca4 cells were injected intraperitoneally (ip) to 16-month-old mice. - / -Mice were given vehicle or zoledronic acid (ZA, 5 μg / kg) three times a week for 8 weeks. At the end of the study, the eyeballs were enucleated and the cornea, lens, vitreous, and retina were removed. 4-6 relaxing incisions were made in the eyecup to generate RPE flat mounts and fixed with 4% paraformaldehyde. The flat mounts were immunostained with antibodies against ionized calcium binding adapter molecule 1 (Iba1), a protein specifically expressed by microglia. Images were captured using a Nikon spinning disk confocal microscope, and the number of microglia in each flat mount was quantified. Figure 7 The results shown in the literature indicate that low-dose zoledronic acid reduces the incidence of Abca4 in 18-month-olds - / - Subretinal microglia in mice. Photoreceptor degeneration

[0088] After 8 weeks of vehicle or ZA administration as described above, electroretinograms were recorded using the Celeris system (Diagnosys LLC, Earle, MA). - / Retinal function of mice. 0.158 to 50 cd.s / m 2 Scotopic (dark-adapted) ERG responses were measured at flash intensities of 1.5 to 2.5 cd.s / m. The a-wave amplitude, a measure of photoreceptor function, was calculated and plotted as a function of flash intensity. 2 ) recorded the c-wave, which is a measure of RPE health and function. Figure 8 The results shown in the literature suggest that low-dose zoledronic acid prevents the development of Abca4 - / - Photoreceptor degeneration and visual deficits in mice. Example 3. In vitro treatment of RPE cells with adiprolon

[0089] Highly differentiated polarized primary RPE cultures were established from freshly harvested porcine eyes as described by Toops et al., 2014. To induce complement-mediated mitochondrial damage, RPE monolayers were treated with the lipofuscin diretinoid A2E, which impairs the mechanisms that protect RPE from complement activation (Tan et al., 2016). RPE cells were exposed to active complement components (10% normal human serum, NHS, for 10 min at 37°C) to induce complement activation on the surface of RPE cells. We have previously shown that this leads to mitochondrial fragmentation in RPE (Tan et al., 2016; La Cunza et al., 2021). Following NHS exposure, cells were treated with 1 μM adipolon (ADP) for 3 h. RPE mitochondria were labeled with MitoTracker Deep Red, and cells were imaged live on a Nikon spinning disk confocal microscope as we have described (Tan et al., 2016; La Cunza et al., 2021). Mitochondrial surfaces were reconstructed from 4D images using Imaris (Bitplane). Fig. 9 The results shown in indicate that adiprolon prevents complement-mediated mitochondrial fragmentation in RPE in vitro.

[0090] In in vitro studies on hepatocytes, cardiomyocytes, pancreatic cancer cell lines, etc., adiprolon was used at a dose of about 25 μM for 24-48 hours (range 10-50 μM, 25 μM was effective). Fig. 9 The experimental results described in were obtained using 1 μM adiprolon for 3 hours. We also used 0.01 μM (currently displayed data). These data show that a 10-1000-fold lower dose (0.01-1 μM) effectively reduces ceramide in primary RPE cultures and prevents complement-induced mitochondrial damage in a shorter time (3 hours).

[0091] All patents, patent applications and publications cited in this specification are incorporated herein by reference, as if each independent patent application or publication specifically and individually indicated to be incorporated by reference. The disclosed embodiments are for illustrative purposes only, not for limiting purposes. Although the present invention has been described with reference to the embodiments described therein, it will be appreciated by those skilled in the art that the structure and elements of the present invention may be modified without departing from the overall spirit and scope of the present invention.

Claims

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

2. The method of claim 1, wherein: Such retinal diseases are disorders mediated by the accumulation of lipofuscin in RPE cells.

3. The method of claim 1, wherein: The retinal disease is Stargardt's macular dystrophy.

4. The method of claim 1, wherein: The retinal disease is dry age-related macular degeneration.

5. The method of claim 1, wherein: The retinal disease is selected from the group consisting of neuronal ceroid lipofuscinosis, Batten disease, Bitt's crystalline dystrophy, Niemann-Pick disease type C, Dohlmann's honeycomb dystrophy, Farber disease, and Best's vitelliform macular dystrophy.

6. The method according to any one of claims 1 to 5, wherein: The drug composition comprises or is incorporated into an implant; a drug eluting device, structure or material; a polymer drug eluting wafer; an injectable hydrogel; or an implantable hydrogel scaffold.

7. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered via an intravitreal implant to deliver a dose of 50 ng / day to 50 μg / day to the eye.

8. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered to the eye as an eye drop or suspension, or an eye ointment or gel at a dose of 50 ng / day to 50 μg / day.

9. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered to the eye by suprachoroidal injection at 50 ng / day to 50 μg / day.

10. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered systemically to provide a dose of 0.001 to 2.0 mg.

11. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered systemically to provide a dose of 0.01 to 2.0 mg.

12. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered systemically in a dose of 0.3 to 0.5 mg.

13. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is systemically administered at a dose of 0.3 to 0.5 mg, 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.

14. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered systemically at a dose of 100 ng to 10 μg per kg body weight, a dose of 1.0 to 7.0 μg per kg body weight, or a dose of about 5.0 μg per kg body weight.

15. The method according to any one of claims 1 to 5, wherein: Zoledronic acid or a derivative is administered systemically in an amount of 0.1, 0.2, 0.3, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 or 10.0 μg per kg body weight.

16. The method according to any one of claims 1 to 5, wherein: The administration is performed by any of the routes including intravenous delivery, intramuscular delivery, intraperitoneal delivery or subcutaneous delivery.

17. The method according to any one of claims 1 to 16, 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, once every other day, once a day, twice a day, and three times a day.

18. The method according to any one of claims 1 to 17, wherein: The pharmaceutical composition comprises zoledronic acid or a derivative thereof and any one of an excipient, a carrier, a diluent, a release formulation, a drug delivery or drug targeting carrier and other active therapeutic agents.

19. The method of claim 18, wherein: The pharmaceutical composition comprises an adiponectin 1 receptor agonist.

20. The method of claim 19, wherein: The adiponectin 1 receptor agonist includes adiprolon.

21. The method of any one of claims 1 to 17, wherein: Zoledronic acid was co-administered with an adiponectin 1 receptor agonist.

22. An intravitreal implant comprising zoledronic acid, which is loaded with 0.001 to 0.3 mg zoledronic acid.

23. The intravitreal implant of claim 22, wherein: The implant is loaded with 0.005 to 2.5 mg zoledronic acid.

24. A topical ophthalmic preparation comprising 0.001-0.05 mg / dose of zoledronic acid.

25. The topical ophthalmic formulation of claim 24 comprising 0.005-0.05 mg / dose of zoledronic acid.

26. An injectable ophthalmic preparation comprising zoledronic acid at a concentration of 1 μg / ml to 10 mg / ml.

27. The injectable ophthalmic preparation of claim 26, comprising zoledronic acid at a concentration of 10 μg / ml to 1 mg / ml.

28. The injectable ophthalmic preparation of claim 26 or 27, wherein The ophthalmic formulation comprises zoledronic acid coupled to a dendrimer or formulated as nanoparticles.

29. A method of treating a retinal disease in a subject in need of treatment for the retinal disease, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising adiprolon.

30. A method of treating a retinal disease in a subject in need of treatment for the retinal disease 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.

31. The method of claim 29 or 30, wherein: Such retinal diseases are disorders mediated by the accumulation of lipofuscin in RPE cells.

32. The method of claim 29 or 30, wherein: The retinal disease is Stargardt's macular dystrophy.

33. The method of claim 29 or 30, wherein: The retinal disease is dry age-related macular degeneration.

34. The method of claim 29 or 30, wherein: The retinal disease is selected from the group consisting of neuronal ceroid lipofuscinosis, Batten disease, Bitt's crystalline dystrophy, Niemann-Pick disease type C, Dohlmann's honeycomb dystrophy, Farber disease, and Best's vitelliform macular dystrophy.

35. The method of any one of claims 29 to 34, wherein: Adipron or the bisphosphonate ASM / FDPS inhibitor is administered at a dose of 0.001 to 2.0 mg, a dose of 0.1 to 1.0 mg, or a dose of 0.3 to 0.5 mg.

36. The method of any one of claims 29-34, wherein adiprolon or a bisphosphonate ASM / FDPS 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 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.

37. The method of any one of claims 29 to 34, wherein: Adipron or a bisphosphonate ASM / FDPS inhibitor is administered at a dose of 100 ng to 10 μg per kg body weight, a dose of 1.0 to 7.0 μg per kg body weight, or a dose of about 5.0 μg per kg body weight.

38. The method of any one of claims 29 to 34, wherein: Adiplon or a bisphosphonate ASM / FDPS inhibitor is administered at a dose selected from the group consisting of 0.1, 0.2, 0.3, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0 μg / kg body weight.

39. The method of any one of claims 29 to 38, 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, once every other day, once a day, twice a day, and three times a day.

40. The method of any one of claims 29 to 39, wherein: Administration to the eye is by any of the following routes: systemic delivery; local delivery; intravenous delivery; intramuscular delivery; intraperitoneal delivery; local delivery; subcutaneous delivery; intraocular delivery; and topical delivery.

41. The method of any one of claims 29 to 39, wherein: The pharmaceutical composition comprises one or more bisphosphonate ASM / FDPS inhibitors or adiprolon and any one of an excipient, carrier, diluent, release formulation, drug delivery or drug targeting carrier and other active therapeutic agents.

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